An Embryo-fetal Development Study of Thymosin beta 4 ...

FINAL REPORT

Testing Facility Study No. 20024504

An Embryo-fetal Development Study of Thymosin beta 4 (formulated as RGN-352) Administered by Intravenous Injection in Rats

SPONSOR: RegeneRx Biopharmaceuticals, Inc. 15245 Shady Grove Road, Suite 470

Rockville, MD 20850 United States

TESTING FACILITY: Charles River Laboratories

Preclinical Services, Pennsylvania (PCS-PA) 905 Sheehy Drive

Horsham, PA 19044 United States

13 February 2013

of 208

Final Report Testing Facility Study No. 20024504

TABLE OF CONTENTS

1. LIST OF FIGURES .................................................................................................................4

2. LIST OF TABLES ..................................................................................................................5

3. LIST OF APPENDICES .........................................................................................................6

4. COMPLIANCE STATEMENT ..............................................................................................7

5. QUALITY ASSURANCE STATEMENT ..............................................................................8

6. RESPONSIBLE PERSONNEL ...............................................................................................9 6.1. Testing Facility ..............................................................................................................9 6.2. Principal Investigators (PI) at Testing Facility ..............................................................9

7. SUMMARY ..........................................................................................................................10

8. INTRODUCTION .................................................................................................................11

9. MATERIALS AND METHODS ..........................................................................................11 9.1. Test and Control Articles .............................................................................................11 9.1.1. Test Article...................................................................................................................11 9.1.2. Control Article .............................................................................................................11 9.2. Test and Control Article Characterization ...................................................................12 9.3. Reserve Samples ..........................................................................................................12 9.4. Test and Control Article Inventory and Disposition ....................................................12 9.5. Safety ...........................................................................................................................12 9.6. Dose Formulation and Analysis ...................................................................................12 9.6.1. Preparation of Control Article .....................................................................................12 9.6.2. Preparation of Test Article ...........................................................................................12 9.6.3. Sample Collection and Analysis ..................................................................................13 9.7. Test System ..................................................................................................................14 9.7.1. Receipt .........................................................................................................................14 9.7.2. Justification for Test System and Number of Animals ................................................14 9.7.3. Animal Identification ...................................................................................................14 9.7.4. Environmental Acclimation .........................................................................................14 9.7.5. Mating ..........................................................................................................................14 9.7.6. Selection, Assignment, and Replacement of Animals .................................................14 9.7.7. Disposition ...................................................................................................................14 9.7.8. Husbandry ....................................................................................................................14 9.8. Experimental Design ....................................................................................................16 9.8.1. Administration of Test and Control Articles ...............................................................16 9.8.2. Justification of Route and Dose Levels .......................................................................16 9.9. In-life Procedures, Observations, and Measurements ..................................................17 9.9.1. Viability Checks ...........................................................................................................17 9.9.2. Clinical Observations ...................................................................................................17 9.9.3. Body Weights ...............................................................................................................17 9.9.4. Food Consumption .......................................................................................................17 9.10. Terminal Procedures ....................................................................................................17 9.10.1. Method of Euthanasia ..................................................................................................17

Final Report Page 2Testing Facility Study No. 20024504


9.10.2. Scheduled Euthanasia ..................................................................................................18 9.10.3. Necropsy ......................................................................................................................18 9.10.4. Tissue Collection and Preservation ..............................................................................18 9.10.5. Fetal Examinations.......................................................................................................19

10. COMPUTERIZED SYSTEMS .............................................................................................19

11. STATISTICAL ANALYSIS .................................................................................................20

12. RETENTION OF RECORDS, SAMPLES, AND SPECIMENS .........................................21

13. RESULTS ..............................................................................................................................22 13.1. Dose Formulation Analyses .........................................................................................22 13.2. Mortality, Clinical and Necropsy Observations...........................................................22 13.3. Maternal Body Weights and Body Weight Changes ...................................................22 13.4. Maternal Food Consumption .......................................................................................22 13.5. Caesarean-Sectioning and Litter Observations ............................................................22 13.6. Fetal Alterations ...........................................................................................................23 13.7. Summary of Fetal Alterations ......................................................................................23 13.7.1. Fetal Gross External Alterations ..................................................................................23 13.7.2. Fetal Soft Tissue Alterations ........................................................................................24 13.7.3. Fetal Skeletal Alterations .............................................................................................24

14. CONCLUSION .....................................................................................................................26

15. REPORT APPROVAL ..........................................................................................................27

16. SCIENTIFIC REPORT REVIEW .........................................................................................28

17. REFERENCES ......................................................................................................................29



1. LIST OF FIGURES

Figure 1 Maternal Body Weights ........................................................................................30



2. LIST OF TABLES

Table 1 Clinical and Necropsy Observations - Summary ..................................................32

Table 2 Maternal Body Weights - Summary .....................................................................34

Table 3 Maternal Body Weight Changes - Summary ........................................................36

Table 4 Maternal Absolute Food Consumption Values (g/day) - Summary .....................38

Table 5 Maternal Relative Food Consumption Values (g/kg/day) - Summary .................40

Table 6 Caesarean-Sectioning Observations - Summary ...................................................42

Table 7 Litter Observations (Caesarean-Delivered Fetuses) - Summary ..........................45

Table 8 Fetal Alterations - Caesarean-Delivered Live Fetuses (Day 21 of Gestation) - Summary ...........................................................................................47

Table 9 Fetal Gross External Alterations - Caesarean-Delivered Live Fetuses (Day 21 of Gestation) - Summary ............................................................49

Table 10 Fetal Soft Tissue Alterations - Caesaraean-Delivered Live Fetuses (Day 21 of Gestation) - Summary ............................................................51

Table 11 Fetal Skeletal Alterations - Caesarean-Delivered Live Fetuses (Day 21 of Gestation) - Summary ...........................................................................................53

Table 12 Fetal Ossification Sites - Caesarean-Delivered Live Fetuses (Day 21 of Gestation) - Summary ...........................................................................................57



3. LIST OF APPENDICES

Appendix 1 Protocol and Deviations ........................................................................................59

Appendix 2 Test and Control Article Characterization ............................................................87

Appendix 3 Dose Formulation Analysis Report .......................................................................92

Appendix 4 Clinical Observations - Individual Data .............................................................135

Appendix 5 Maternal Body Weights - Individual Data ..........................................................140

Appendix 6 Maternal Food Consumption Values - Individual Data ......................................149

Appendix 7 Necropsy Observations - Individual Data ...........................................................154

Appendix 8 Caesarean-Sectioning Observations - Individual Data .......................................159

Appendix 9 Litter Observations (Caesarean-Delivered Fetuses) - Individual Data ...............164

Appendix 10 Fetal Sex, Vital Status and Body Weight - Individual Data ...............................169

Appendix 11 Fetal Alterations - Individual Data .....................................................................178

Appendix 12 Historical Control Data .......................................................................................191

Appendix 13 Tabulated Summary for The Common Technical Document (CTD) .................206



4. COMPLIANCE STATEMENT The study was performed in accordance with the U.S. Department of Health and Human Services, Food and Drug Administration (FDA), United States Code of Federal Regulations, Title 21, Part 58: Good Laboratory Practice for Nonclinical Laboratory Studies and as accepted by Regulatory Authorities throughout the European Community (OECD Principles of Good Laboratory Practice) and Japan (MHLW).

One exception from the FDA Good Laboratory Practice (GLP) for Nonclinical Laboratory Studies regulation occurred. Analyses conducted to support the information cited in the Certificate of Analysis for the test and control articles were conducted in accordance with current FDA Good Manufacturing Practice regulations. The outcome or interpretation of the study was not adversely affected by this exception, because the FDA GLP requires that “The identity, strength, purity, and composition or other characteristics which will appropriately define the test or control article shall be determined for each batch and shall be documented [58.105(a)].” This requirement is met, therefore there is no effect on the study.

This study was conducted in accordance with the procedures described herein. All deviations authorized/acknowledged by the Study Director are documented in the Study Records. The report represents an accurate and complete record of the results obtained.

There were no deviations from the above regulations that affected the overall integrity of the study or the interpretation of the study results and conclusions.



6. RESPONSIBLE PERSONNEL

6.1. Testing Facility Study Director Valerie A. Sharper, MS

Principal Research Scientist Address as cited for Testing Facility

Testing Facility Management Alan M. Hoberman, PhD, DABT, Fellow ATS Executive Director, Site Operations and Toxicology Address as cited for Testing Facility

Director of Operations Matthew J. Vaneman, BS

Senior Manager Technical Operations Joseph W. Lech, BS, LAT

Senior Manager Technical Resources James Yeager, BS, LAT

Senior Manager Laboratory Sciences Julian Gulbinski, III, BS, MBA

6.2. Principal Investigators (PI) at Testing Facility Dose Formulation Analysis Jason Sarsoza, BSc

Scientist II Address as cited for Testing Facility



7. SUMMARY The objectives of this study were to detect adverse effects of Thymosin beta 4, formulated as Tβ4 Injectable Solution (RGN-352), on pregnant Crl:CD(SD) rats and development of the embryo and fetus consequent to exposure of the female from implantation to closure of the hard palate. This study was designed to evaluate ICH Harmonised Tripartite Guideline stages C to D of the reproductive process.

One hundred Crl:CD(SD) rats were randomly assigned to four dose groups, 25 rats per group. Rats were administered the control article or test article formulation via intravenous bolus injection at doses of 0, 20, 60 and 180 mg/kg/day on Days 7 through 17 of presumed gestation (DGs 7 through 17). The study design was as follows:

Text Table 1 Experimental Design

Group No. Test Material

Dose Level (mg/kg)

Concentration (mg/mL)

Dose Volume (mL/kg) No. of Rats

1 Control Article 0 0 1.8 25 2

RGN-352 20 100 0.2 25

3 60 100 0.6 25 4 180 100 1.8 25

The following parameters and end points were evaluated in this study: viability, clinical signs, body weights, body weight changes, food consumption, ovarian and uterine parameters, fetal examinations (body weight, sex, gross external, visceral and skeletal alterations) and gross necropsy observations.

All rats survived until scheduled euthanasia. All clinical observations were considered unrelated to Tβ4 Injectable Solution and all rats appeared normal at necropsy. Body weights, body weight gains and absolute and relative food consumption values were unaffected by dosages of Tβ4 Injectable Solution as high as 180 mg/kg/day.

No Caesarean-sectioning or litter parameters were affected by dosages of Tβ4 Injectable Solution as high as 180 mg/kg/day. No gross external, soft tissue or skeletal fetal alterations (malformations or variations) were caused by Tβ4 Injectable Solution. Fetal ossification site averages were comparable among the four dose groups, and no toxicologically important differences occurred.

On the basis of these data, the maternal no-observable-effect level (NOEL) of Tβ4 Injectable Solution is 180 mg/kg/day. The developmental NOEL is also 180 mg/kg/day. No adverse effects on maternal rats or embryo-fetal development were observed at the highest dosage tested.



8. INTRODUCTION The objectives of this study were to detect adverse effects of Thymosin beta 4, formulated as Tβ4 Injectable Solution (RGN-352), on pregnant Crl:CD(SD) rats and development of the embryo and fetus consequent to exposure of the female from implantation to closure of the hard palate. This study was designed to evaluate ICH Harmonised Tripartite Guideline stages C to D of the reproductive process.

The design of this study was based on the study objectives, the overall product development strategy for the test article, and based on ICH Harmonised Tripartite Guidelines1,2.

The Study Director signed the protocol on 01 May 2012, and dosing was initiated on 15 May 2012. The in-life phase of the study was completed on 01 Jun 2012. The experimental start date was 01 May 2012, and the experimental completion date was 11 Jul 2012. The study protocol and deviations are presented in Appendix 1.

9. MATERIALS AND METHODS

9.1. Test and Control Articles

9.1.1. Test Article Identification: Thymosin beta 4 (Tβ4) formulated as RGN-352 (Tβ4 Injectable

Solution)

Lot No.: 330-05-001

Receipt Date: 22 Aug 2011

Expiration Date: 01 Dec 2012

Physical Description: Clear, colorless liquid, 2.25 mg/vial

Concentration: 100 mg Tβ4/mL

Storage Conditions: Refrigerated (2°C to 8°C)

Supplier: Sponsor

9.1.2. Control Article Identification: RGN-352 Placebo (Vehicle for RGN-352)

Batch (Lot) No.: 330-06-001

Receipt Date: 22 Aug 2011

Expiration Date: 27 Aug 2012

Physical Description: Clear, colorless liquid, 5.5 mL/vial


Supplier: Sponsor



9.2. Test and Control Article Characterization The Sponsor provided to the Testing Facility documentation of the identity, strength, purity, composition and stability for the test and control articles. Certificates of Analysis were provided to the Testing Facility and are presented in Appendix 2.

The Sponsor has appropriate documentation on file concerning the method of synthesis, fabrication or derivation of the test and control articles, and this information is available to the appropriate regulatory agencies should it be requested.

9.3. Reserve Samples For each batch (lot) of test and control article, a reserve sample (1 vial) was collected and maintained under the appropriate storage conditions by the Testing Facility.

9.4. Test and Control Article Inventory and Disposition Records of the receipt, distribution, storage and disposition of test and control articles (including empty containers) were maintained. With the exception of reserve samples, all unused test and control articles were retained for use in future studies. All empty containers were maintained for the duration of the study. Any residual volumes were discarded before issuance of the Final Report.

9.5. Safety The following safety instructions applied to this study:

Gloves, dust-mist/HEPA-filtered mask, appropriate eye protection and uniform/lab coat. Tyvek® sleeves were worn during dose administration.

The Material Safety Data Sheet (MSDS) is maintained in the raw data.

9.6. Dose Formulation and Analysis

9.6.1. Preparation of Control Article The control article, RGN-352 Placebo, was administered as received. The control article was dispensed daily for administration to Group 1 control animals. The control article was removed from the refrigerator at least 30 minutes before dosing to allow it to equilibrate to room temperature.

9.6.2. Preparation of Test Article

The test article, Tβ4 Injectable Solution (RGN-352), was administered as received. The test article was dispensed daily for administration to Groups 2, 3 and 4. The test article was removed from the refrigerator for at least 30 minutes before dosing to allow it to equilibrate to room temperature.



9.6.3. Sample Collection and Analysis Dose formulation samples were collected for analysis as indicated in Text Table 2.

Text Table 2 Dose Formulation Sample Collection Schedule

Interval Concentration Homogeneity Stability Before First Administration 0 and 100 mg/mL N/A N/A Last Day of Administrationa 0 and 100 mg/mL N/A N/A N/A = Not applicable. All samples analyzed were transferred (ambient conditions) to the analytical laboratory at the Testing Facility (see Appendix 1, Deviations). Upon receipt at the analytical laboratory, the samples were stored refrigerated (2°C to 8°C) until analysis.

Any residual analytical samples (and test article used in analysis) were discarded before issue of the Final Report.

9.6.3.1. Analytical Method Analyses described below were performed by HPLC-UV using validated analytical procedure TYMN03 - Analytical Procedure for the Analysis of Thymosin Beta 4 in RGN-352 Placebo Dose Formulation by HPLC-UV; validated under PCS-PA Protocol No. 20019481.

9.6.3.2. Concentration Analysis Samples (one vial each) of the test and control articles were taken before the first dose administration and on the last day of dose administration and transferred to the analytical laboratory at the Testing Facility for analysis; another set of vials was retained as backup samples on the last day of dose administration and stored refrigerated (2°C to 8°C) (see Appendix 1, Deviations). Concentration results were considered acceptable if mean sample concentration results were within or equal to ± 10% of theoretical concentration. Each individual sample concentration result was considered acceptable if it were within or equal to ± 15%. After acceptance of the analytical results, backup samples (unopened vials) were retained for use on future studies.

9.6.3.3. Homogeneity Analysis The test article Tβ4, formulated and vialed as RGN-352 (100 mg/mL), yields a sterile aqueous, homogeneous solution. Therefore, homogeneity analysis of the test article was not conducted. The test article was not mixed with the placebo (RGN-352 excipients minus Tβ4) in this study.

9.6.3.4. Stability Analysis

Stability of the formulated test article for the duration of the study was confirmed by comparison of the results of the concentration analysis conducted for samples collected before the first dose administration and samples collected on the last day of administration.



9.7. Test System

9.7.1. Receipt One-hundred and twenty virgin female Crl:CD(SD) Sprague Dawley rats were received from Charles River Laboratories, Kingston, NY. The rats were approximately 66 days of age upon arrival at the Testing Facility. The body weight range was 204 g to 255 g on the day after arrival and 227 g to 258 g at randomization and assignment to study.

9.7.2. Justification for Test System and Number of Animals The test system was selected because: 1) it is a standard species accepted for use in embryo-fetal development studies; 2) this species and strain has been demonstrated to be sensitive to developmental toxicants; and 3) historical data and experience exist at the Testing Facility.

The number of animals chosen for this study was the smallest number considered necessary to provide the minimum number of pregnancies recommended by the applicable guidelines.

9.7.3. Animal Identification Female rats were permanently identified using Monel® self-piercing ear tags. Male rats were permanently identified by tattoo and given permanent unique identification numbers upon assignment to the Testing Facility's breeder male rat population. Female rats were assigned temporary numbers at receipt and given unique permanent identification numbers when assigned to the study on the basis of DG 0 body weights.

9.7.4. Environmental Acclimation After receipt at the Testing Facility, the rats were acclimated for 5 days before cohabitation.

9.7.5. Mating Female rats were mated with breeder male rats of the same source and strain, 1 male per female rat. The cohabitation period consisted of a maximum of 5 days. Female rats were examined each day of the cohabitation period and those with spermatozoa observed in a smear of the vaginal contents and/or a copulatory plug observed in situ were considered mated and to be DG 0.

9.7.6. Selection, Assignment, and Replacement of Animals

One-hundred healthy mated female rats, 25 per dose group, were assigned to groups using a computer-based weight-ordered randomization procedure, based on DG 0 body weights.

9.7.7. Disposition The disposition of all animals was documented in the study records. All remaining rats not selected for study and with no confirmed date of mating were euthanized.

9.7.8. Husbandry All cage sizes and housing conditions are in compliance with the Guide for the Care and Use of Laboratory Animals.3



9.7.8.1. Housing The rats were individually housed in stainless steel, wire-bottomed cages, except during the cohabitation period. During cohabitation, each pair of rats was housed in the male rat's nesting box. No nesting materials were supplied to female rats that were individually housed following completion of the cohabitation period because the female rats were euthanized before parturition was expected.

9.7.8.2. Environmental Conditions The study rooms were maintained under conditions of positive airflow relative to a hallway and independently supplied with a minimum of ten changes per hour of 100% fresh air that had been passed through 99.97% HEPA filters. Room temperature and humidity were monitored constantly throughout the study. Room temperature was targeted at 64°F to 79°F (18°C to 26°C); relative humidity was targeted at 30% to 70% (See Appendix 1, Deviations). An automatically controlled 12-hour light:12-hour dark fluorescent light cycle was maintained. Each dark period began at 1900 hours (± 30 minutes).

9.7.8.3. Food Rats were given Certified Rodent Diet® #5002 (PMI® Nutrition International) available ad libitum from individual feeders.

Analyses were routinely performed by the food supplier. No contaminants at levels exceeding the maximum concentration limits for certified food or deviations from expected nutritional requirements were detected by these analyses. Copies of the results of the food analyses are available in the raw data.

The Study Director was not aware of any potential contaminants likely to be present in the certified diet that would have interfered with the results of this study. Therefore, no analyses other than those routinely performed by the food supplier or those mentioned in this report were conducted.

9.7.8.4. Water Local water (Horsham, PA), that had been processed by passage through a reverse osmosis membrane (R.O. water), was available to the rats ad libitum from an automatic watering access system and/or water bottles attached to the cages. Chlorine was added to the processed water as a bacteriostat.

The processed water is analyzed twice annually for possible chemical contamination (Lancaster Laboratories, Lancaster, PA) and monthly for possible bacterial contamination (QC Laboratories, Southampton, PA).

The Study Director was not aware of any potential contaminants likely to be present in the drinking water at levels that would interfere with the results of this study. Therefore, no analyses other than those mentioned in this report were conducted.

9.7.8.5. Animal Enrichment Chewable Nylabones® were supplied to all rats during the course of the study, except during cohabitation. Analyses for possible contamination were conducted on each lot of Nylabones®.



The Study Director was not aware of any potential contaminants likely to be present in the enrichment products at levels that would have interfered with the results of this study. Therefore, no analyses other than those routinely performed or those mentioned in this report were conducted.

9.7.8.6. Veterinary Care Veterinary care was available throughout the course of the study; however, no veterinary examinations or medical treatments were administered during the course of the study.

9.8. Experimental Design The experimental design is presented in Text Table 3, below.

Text Table 3 Experimental Design


Dose Level (mg/kg/day)


Dose Volume (mL/kg) Rat Numbers

1 Control Article 0 0 1.8 4001 - 4025 2

RGN-352 20 100 0.2 4026 - 4050

3 60 100 0.6 4051 - 4075 4 180 100 1.8 4076 - 4100

9.8.1. Administration of Test and Control Articles Female rats were administered the test article or the control article by once daily intravenous bolus injection via the lateral tail vein on DGs 7 through 17. The actual dose volumes were adjusted based on the most recently recorded body weight and administered at approximately the same time each day.

9.8.2. Justification of Route and Dose Levels The intravenous route was selected for use because it is one of the proposed systemic routes for clinical use.

Dose range selections for the test article in this study encompass the dose level selections planned for RGN-352 in future clinical trials. The 180 mg/kg dose level is 6 times higher than the highest dose anticipated for administration in man. The low dose level of 20 mg/kg is at least 3 times higher than the lowest dose anticipated for administration in man. If an NOAEL cannot be established for the mated female and the development of the embryo and fetus from implantation to closure of the hard palate, then at least a 6-fold margin of safety would be established based on the highest dose planned for RGN-352 in man.

Dosages were also selected on the basis of a dosage-range developmental toxicity study (Testing Facility Study No. 20024503). In that study, RGN-352 was administered once daily by intravenous bolus injection from DGs 7 to 17 at levels of 10, 30, 100 and 300 mg/kg/day. There were no test article-related adverse clinical observations and body weight gains and food consumption values were comparable among the five dose groups for the entire dose period. All



Caesarean-sectioning and litter parameters were comparable among the five dose groups and there were no fetal gross external alterations.

9.9. In-life Procedures, Observations, and Measurements The in-life procedures, observations, and measurements listed below were performed for all rats.

9.9.1. Viability Checks The rats were assessed for viability at least twice daily during the study (See Appendix 1, Deviations).

9.9.2. Clinical Observations The rats were observed for general appearance at least twice weekly during the acclimation period, daily before administration during the dose period and once daily during the postdose period.

Postdose observations were recorded between 1 and 2 hours after dose administration.

9.9.3. Body Weights Body weights were recorded at least twice weekly during the acclimation period, on DG 0, and daily during the dose and postdose periods.

9.9.4. Food Consumption Food consumption values were recorded on DGs 0, 7, 10, 12, 15, 18, and 21.

9.10. Terminal Procedures Terminal procedures are summarized in Text Table 4.

Text Table 4 Terminal Procedures

Group No.

No. of Animals

Scheduled Euthanasia

Day

Necropsy Procedures

Histology Histopathology

Ovarian/ Uterine

Examination NecropsyTissue

CollectionOrgan

Weights 1 25

DG 21 Full Exam X X -

- - 2 25 - -

3 25 - -

4 25 - - X = Procedure conducted; - = Not applicable. DG = Day of Presumed Gestation

9.10.1. Method of Euthanasia Rats were euthanized by carbon dioxide asphyxiation. Live fetuses were euthanized by an intraperitoneal injection of sodium pentobarbital.



9.10.2. Scheduled Euthanasia On DG 21, all rats were Caesarean-sectioned and examined for gross lesions as described in Section 9.10.2.1 (Ovarian and Uterine Examinations) and Section 9.10.3 (Necropsy).

9.10.2.1. Ovarian and Uterine Examinations The reproductive tract was dissected from the abdominal cavity. The uterus was opened, and the contents were examined. The fetuses were removed from the uterus and placed in individual containers.

The ovaries and uterus of each rat were examined for number and distribution of corpora lutea, implantation sites, placentae (size, color or shape), live and dead fetuses, and early and late resorptions. An early resorption was defined as one in which organogenesis was not grossly evident. A late resorption was defined as one in which the occurrence of organogenesis was grossly evident. A live fetus was defined as a term fetus that responded to stimuli. Nonresponding term fetuses were considered to be dead. Dead fetuses and late resorptions were differentiated by the degree of autolysis present; marked to extreme autolysis indicated that the fetus was a late resorption.

Uteri of apparently nonpregnant rats were examined while being pressed between glass plates to confirm the absence of implantation sites. Uteri and ovaries of apparently nonpregnant rats were retained in 10% neutral buffered formalin and were discarded when authorized by the Study Director.

9.10.3. Necropsy Rats were subjected to a gross necropsy examination, which included an evaluation of the thoracic, abdominal, and pelvic cavities with their associated organs and tissues.

9.10.4. Tissue Collection and Preservation Representative samples of the tissues identified in Text Table 5 were collected from all animals and preserved in 10% neutral buffered formalin, unless otherwise indicated.

A table of random units was used to select one control group rat from which all tissues examined at necropsy were retained, in order to provide control tissues for any possible histopathological evaluations of gross lesions.

Text Table 5 Tissue Collection and Preservation

Tissue Collected Comment Animal identification (ear tag) X All rats with retained tissues.

Cervix X Collected with the uterus. All nonpregnant rats. Ovaries X All nonpregnant rats. Uterus X All nonpregnant rats.

X = Procedure conducted



9.10.5. Fetal Examinations Representative photographs of visceral and skeletal abnormalities were taken. Photographs were not included in the report, but were retained as electronic images and archived with the raw data.

Fetuses were examined for sex and external abnormalities. Dead fetuses and late resorptions were examined for sex and external abnormalities to the extent possible.

The body weight of each fetus was recorded. Fetuses were individually identified with the study number, litter number and uterine distribution.

Approximately one-half of the fetuses in each litter were examined for visceral abnormalities by using a modification of the microdissection technique of Staples4. Each fetus was fixed in Bouin's solution, and the heads were subsequently examined by free-hand sectioning;5 head sections were stored in alcohol (See Appendix 1, Deviations). The decapitated carcasses were not retained. The remaining fetuses (approximately one-half of the fetuses in each litter) were examined for skeletal abnormalities after staining with alizarin red S.6 (See Appendix 1, Deviations). Following examination, skeletal preparations were retained in glycerin with thymol added as a preservative (See Appendix 1, Deviations).

10. COMPUTERIZED SYSTEMS Critical computerized systems used in the study are listed below or presented in the appropriate Phase Report. All computerized systems used in the conduct of this study have been validated; when a particular system has not satisfied all requirements, appropriate administrative and procedural controls were implemented to assure the quality and integrity of data.

Text Table 6 Critical Computerized Systems

System Name Version No. Description of Data Collected and/or Analyzed Dispense 7.0.3 Test article/control article receipt

Argus Automated Data Collection and Management

System 13.11.18.9 Clinical observations, body weights, food consumption,

Caesarean-sectioning observations, statistical analyses

Vivarium Temperature and Relative Humidity Monitoring System

2.0 Study room temperature and humidity

Microsoft® Excel 2007 Pre- and postimplantation loss (report summary table) Quattro Pro 8 Body weight graph

The SAS System 6.12 Statistical analyses



11. STATISTICAL ANALYSIS Averages and percentages were calculated. Litter values were used where appropriate.



Clinical observations and other proportional data were analyzed using the Variance Test for Homogeneity of the Binomial Distribution.7

Continuous data (e.g., maternal body weights, body weight changes, food consumption values and litter averages for percent male fetuses, percent resorbed conceptuses, fetal body weights and fetal anomaly data) were analyzed using Bartlett’s Test of Homogeneity of Variances8 and the Analysis of Variance9, when appropriate [i.e., Bartlett’s Test was not significant (p0.001)]. If the Analysis of Variance was significant (p0.05), Dunnett’s Test10 was used to identify the statistical significance of the individual groups. If the Analysis of Variance was not appropriate [i.e., Bartlett’s Test was significant (p0.001)], the Kruskal-Wallis Test11 was used (75% ties). In cases where the Kruskal-Wallis Test was statistically significant (p0.05), Dunn’s Method of Multiple Comparisons12 was used to identify the statistical significance of the individual groups. If there were greater than 75% ties, Fisher’s Exact Test13 was used to analyze the data.

Count data were evaluated using the procedures described above for the Kruskal-Wallis Test.11

12. RETENTION OF RECORDS, SAMPLES, AND SPECIMENS All study-specific raw data, documentation, protocol, samples, specimens, and final reports from this study were available at the Testing Facility during the study and will be transferred to the Testing Facility archive by no later than the date of Final Report issue. One year after issue of the audited draft report, the Sponsor will be contacted to determine the disposition of materials associated with the study.

All records, samples, specimens, and reports generated from phases or segments performed by Testing Facility-designated subcontractors were sent to the Testing Facility for archiving.



13. RESULTS

13.1. Dose Formulation Analyses (Appendix 3)

Mean measured Tβ4 concentrations, sampled at the start and end of the study, were within the acceptable limits of ± 10% of nominal concentrations. Individual Tβ4 concentrations for all samples were within the acceptable limits of ± 15% of nominal concentrations. There was no test article contamination of the control article.

13.2. Mortality, Clinical and Necropsy Observations (Table 1, Appendix 4 and Appendix 7)

All rats survived until scheduled euthanasia.

All clinical observations were considered unrelated to Tβ4 Injectable Solution because: 1) the incidences were not dose dependent; 2) the observations are common in the laboratory environment; and/or 3) the observations occurred only in one rat in the control group and three different rats in the highest dose group of 180 mg/kg. These clinical observations included portion of tail missing and sparse hair coat on the underside, limbs and/or neck.

All rats appeared normal at necropsy.

13.3. Maternal Body Weights and Body Weight Changes (Figure 1, Table 2, Table 3, and Appendix 5)

Body weights and body weight gains were unaffected by dosages of Tβ4 Injectable Solution as high as 180 mg/kg/day. Average body weights on Day 18 of gestation (DG 18), the day after the last dose, were 99.2%, 100% and 101.9% of the control group value in the 20, 60 and 180 mg/kg/day dose groups, respectively. A statistically significant reduction (p≤0.05) in body weight gain in the 20 mg/kg/day dose group on DGs 10 to 12 as compared to the control group was not considered test article related because it was not dose dependent.

13.4. Maternal Food Consumption (Table 4, Table 5 and Appendix 6)

Absolute and relative food consumption values were unaffected by dosages of Tβ4 Injectable Solution as high as 180 mg/kg/day. All values were comparable among the four dose groups and no statistically significant differences occurred. Absolute food consumption values for the entire dose period (calculated as DGs 7 to 18) were 98.0%, 100.8% and 101.2% of the control group value in the 0, 20, 60 and 180 mg/kg/day dose groups, respectively.

13.5. Caesarean-Sectioning and Litter Observations (Table 6, Table 7, Appendix 8, Appendix 9, and Appendix 10)

Caesarean-sectioning observations were based on 25 (100%), 25 (100%), 25 (100%) and 23 (92.0%) pregnant rats in Groups 1 through 4, respectively.



No Caesarean-sectioning or litter parameters were affected by dosages of Tβ4 Injectable Solution as high as 180 mg/kg/day. The litter averages for corpora lutea, implantations, percent preimplantation loss, litter sizes, live fetuses, early and late resorptions, percent postimplantation loss, percent resorbed conceptuses, and percent live male fetuses were comparable among the four dose groups and did not significantly differ. No dam had a litter consisting of only resorbed conceptuses, and there were no dead fetuses. All placentae appeared normal. Statistically significant increases (p≤0.05 or p≤0.01) in total and male fetal body weights in the 60 mg/kg/day dose group were not considered test article related because they were not dose dependent.

13.6. Fetal Alterations (Table 8 and Appendix 11)

Fetal alterations were defined as: 1) malformations (irreversible changes that occur at low incidences in this species and strain); or 2) variations (common findings in this species and strain and reversible delays or accelerations in development). Litter averages were calculated for specific fetal ossification sites as part of the evaluation of the degree of fetal ossification.

Fetal evaluations were based on 333, 340, 338 and 324 live, DG 21 Caesarean-delivered fetuses in 25, 25, 25, and 23 litters in the 0 (Control), 20, 60 and 180 mg/kg/day dose groups, respectively. Each of these fetuses was examined for gross external alterations. Of these respective fetuses, 161, 164, 161 and 159 fetuses were examined for soft tissue alterations, and 173, 176, 177 and 165 fetuses were examined for skeletal alterations and fetal ossification site averages.

No gross external, soft tissue or skeletal fetal alterations (malformations or variations) were caused by dosages of Tβ4 Injectable Solution as high as 180 mg/kg/day. There were no dose-dependent or significant differences in the litter or fetal incidences of any gross external, soft tissue or skeletal alterations. Fetal ossification site averages were comparable among the four dose groups, and no toxicologically important differences occurred.

13.7. Summary of Fetal Alterations (Table 8, Appendix 10 and Appendix 11)

In Groups 1 through 4, litters with fetuses with alterations numbered 7 (28.0%), 10 (40.0%), 4 (16.0%) and 3 (13.0%), respectively. The numbers of fetuses with any alteration observed were 8 (2.4%), 15 (4.4%), 4 (1.2%) and 6 (1.8%), and the percentages of fetuses with any alteration per litter were 2.7, 4.6, 1.1 and 1.8 in these same respective dose groups. A statistically significant increase (p≤0.01) in the numbers of fetuses with any alteration observed in the 20 mg/kg/day dose group was not considered test article related because it was not dose dependent.

13.7.1. Fetal Gross External Alterations (Table 9 and Appendix 11)

No gross external fetal alterations occurred.



13.7.2. Fetal Soft Tissue Alterations (Table 10 and Appendix 11)

Variations of the vessels occurred in one fetus in each of the control and 20 mg/kg/day dose groups. Fetus 4010-13 in the control group had the aorta pass dorsal to the trachea and esophagus, the innominate artery absent and several vessels (1st left carotid, 2nd right carotid, 3rd right subclavian and the 4th left subclavian) rose in the incorrect order. Fetus 4041-6 in the 20 mg/kg/day dose group had an absent innominate artery. No additional alterations occurred in these fetuses.

13.7.3. Fetal Skeletal Alterations (Table 11 and Appendix 11)

13.7.3.1. Malformations Fetus 4045-1 in the 20 mg/kg/day dose group had a small left eye socket and interrelated vertebral-rib malformations consisting of bifid 11th and 12th thoracic centra and fused left 11th and 12th ribs, and unilateral ossification (left) and small arch (right) of the 5th thoracic vertebra and fused right 4th and 5th ribs. This fetus also had incompletely ossified and fused cervical arches, and asymmetric sternal centra. The litter and fetal incidences of these alterations were within the ranges observed historically at the Testing Facility (Appendix 12).

13.7.3.2. Variations

13.7.3.2.1. Skull One fetus (4087-11) in the 180 mg/kg/day dose group had large nasal-frontal sutures. This fetus also had wavy right 10th and 11th ribs, incompletely ossified right 10th and left 11th ribs, and a thickened right 9th rib.

13.7.3.2.2. Vertebrae Fetus 4045-1 in the 20 mg/kg/day dose group had incompletely ossified and/or fused arches of the 4th and 5th cervical vertebrae. This fetus had additional alterations as previously described.

One fetus in the 20 mg/kg/day dose group (4045-7) had incompletely ossified arches of the 4th and 5th cervical vertebrae as the only alteration. The litter and fetal incidences of this alteration were within the ranges observed historically at the Testing Facility (Appendix 12).

The arch of the 6th cervical vertebra had the appearance of the arch of the 7th cervical vertebra in two fetuses from two different litters in the control group. The litter and fetal incidences of this alteration were within the ranges observed historically at the Testing Facility (Appendix 12).

A bifid centrum in a thoracic vertebra occurred in one fetus in each of the 0 and 60 mg/kg/day dose groups and in four fetuses from different litters in the 20 mg/kg/day dose group. One fetus in the 20 mg/kg/day dose group (4045-1) had additional alterations as previously described. No additional alterations occurred in these fetuses. The litter and fetal incidences of this alteration were within the ranges observed historically at the Testing Facility (Appendix 12).



13.7.3.2.3. Ribs A cervical rib at the 7th cervical vertebra, a common variation in this strain of rat14, was present as the only alteration in one fetus in all four dose groups. The litter and fetal incidences of this alteration were within the ranges observed historically at the Testing Facility (Appendix 12).

Wavy ribs, a reversible delay in ossification15, 16, occurred in 1, 4, 1 and 3 fetuses from 1, 3, 1 and 1 litters in the 0, 20, 60 and 180 mg/kg/day dose groups, respectively. Fetuses 4037-1, -3; 4048-15 and 4049-7 in the control group, 4066-5 in the 60 mg/kg/day dose group and 4087-3, -9 and -11 in the 180 mg/kg/day dose group also had incompletely ossified ribs. Fetus 4087-11 in the 180 mg/kg/day dose group had additional alterations as previously described.

Incompletely ossified ribs occurred in 2, 6, 1 and 4 fetuses from 2, 4, 1 and 2 litters in the 0, 20, 60 and 180 mg/kg/day dose groups, respectively. Additional alterations in these fetuses were described previously.

One fetus (4087-11) in the 180 mg/kg/day dose group had a thickened right 9th rib in addition to wavy and incompletely ossified ribs, as previously described.

Fetus 4087-1 in the 180 mg/kg/day dose group had a short 13th rib as the only alteration. The litter and fetal incidences of this alteration were within the ranges observed historically at the Testing Facility (Appendix 12).

13.7.3.2.4. Sternum Fetus 4045-1 in the 20 mg/kg/day dose group had asymmetric 1st and 2nd sternal centra. This fetus had additional alterations as previously described.

One fetus (4043-3) in the 20 mg/kg/day dose group had duplicated 1st through 3rd sternal centra and xiphoid.

An incompletely ossified 1st sternebra occurred as the only alteration in one fetus in the 20 mg/kg/day dose group.

The litter and fetal incidences of these alterations were within the ranges observed historically at the Testing Facility (Appendix 12).

13.7.3.2.5. Pelvis The pubes were incompletely ossified in one fetus in the 60 mg/k/day dose group. No additional alterations occurred in this fetus.

13.7.3.2.6. Fetal Ossification Site Averages (Table 12)

There were no statistically significant or biologically important differences among the four dose groups in the average numbers of ossification sites per fetus for the hyoid, vertebrae (cervical, thoracic, lumbar, sacral and caudal), ribs, sternum (manubrium, sternal centers and xiphoid), forelimbs (carpals, metacarpals and phalanges) or hindlimb tarsals and metatarsals. Statistically significant increases (p≤0.05 or p≤0.01) in the average number of ossified hindlimb phalanges in the 60 and 180 mg/kg/day dose groups were not considered to be biologically important because: 1) a delay, rather than an acceleration in ossification is the expected effect of a toxicant; and 2) all values were within the ranges observed historically at the Testing Facility (Appendix 12).



14. CONCLUSION On the basis of these data, the maternal no-observable-effect level (NOEL) of Tβ4 Injectable Solution is 180 mg/kg/day. The developmental NOEL is also 180 mg/kg/day. No adverse effects on maternal rats or embryo-fetal development were observed at the highest dosage tested.



17. REFERENCES 1. ICH Harmonised Tripartite Guideline M3 (R2). Nonclinical Safety Studies for the

Conduct of Human Clinical Trials for Pharmaceuticals.

2. ICH Harmonised Tripartite Guideline S5 (R2). Guideline for Industry: detection of toxicity to reproduction for medicinal products & toxicity to male fertility.

3. National Research Council. Guide for the Care and Use of Laboratory Animals. Washington, D.C.: National Academy Press. 1996.

4. Staples RE. Detection of visceral alterations in mammalian fetuses. Teratology 1974;9(3):A37-A38.

5. Wilson JG. Methods for administering agents and detecting malformations in experimental animals. In: Wilson JG, Warkany J, editors. Teratology: principles and techniques. Chicago (IL): University of Chicago Press; 1965. p. 262-77.

6. Staples RE, Schnell VL. Refinements in rapid clearing technic in the KOH-alizarin red S method for fetal bone. Stain Technol 1964;39:61-3.

7. Snedecor GW, Cochran WG. Variance test for homogeneity of the binomial distribution. Statistical methods. 6th Ed. Iowa State University Press, Ames; 1967. p. 240-1.

8. Sokal RR, Rohlf FJ. Bartlett' s test of homogeneity of variances. Biometry: the principles and practice of statistics in biological research. San Francisco (CA): Freeman & Co; 1969. p. 370-1.

9. Snedecor GW, Cochran WG. Analysis of variance. Statistical methods. 6th Ed. Iowa State University Press, Ames; 1967. p. 258-98.

10. Dunnett CW. A multiple comparison procedure for comparing several treatments with a control. J Am Stat Assoc 1955;50:1096-121.

11. Sokal RR, Rohlf FJ. Kruskal-Wallis test. Biometry: the principles and practice of statistics in biological research. San Francisco (CA): Freeman & Co; 1969. p. 388-91.

12. Dunn OJ. Multiple comparisons using rank sums. Technometrics 1964;6(3):241-52.

13. Siegel S. The Fisher’s exact probability test. Nonparametric statistics for the behavioral sciences. New York (NY): McGraw-Hill Co; 1956. p. 96-105.

14. Khera KS. Common fetal aberrations and their teratologic significance: a review. Fund Appl Toxicol 1981;1:13-8.

15. Nishimura M, Iizuka M, Iwaki S, Kast A. Repairability of drug-induced "wavy ribs" in rat offspring. Arzneimittelforschung 1982;32(12):1518-22.

16. Sterz H, Sponer G, Neubert P, Hebold G. A postulated mechanism of beta-sympathomimetic induction of rib and limb anomalies in rat fetuses. Teratology 1985;31(3):401-12.


Revised Draft Report Testing Facility Study No. 20024504

Figure 1 Maternal Body Weights


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MATERNAL BODY WEIGHTSFigure 1

PROTOCOL 20024504: AN EMBRYO-FETAL DEVELOPMENT STUDY OF THYMOSIN BETA 4 (FORMULATED AS RGN-352) BY INTRAVENOUS INJECTION IN RATS

DAY OF GESTATION



Table 1 Clinical and Necropsy Observations - Summary


Revised Draft Report Testing Facility Study No. 20024504 PROTOCOL 20024504: AN EMBRYO-FETAL DEVELOPMENT STUDY OF THYMOSIN BETA 4 (FORMULATED AS RGN-532) BY INTRAVENOUS INJECTION IN RATS TABLE 1 (PAGE 1): CLINICAL AND NECROPSY OBSERVATIONS - SUMMARY ------------------------------------------------------------------------------------------------------------------------------------ GROUP 1 2 3 4 TEST MATERIAL CONTROL ARTICLE RGN-352 RGN-352 RGN-352 DOSE (MG/KG/DAY)a 0 (CONTROL) 20 60 180 ------------------------------------------------------------------------------------------------------------------------------------ MAXIMUM POSSIBLE INCIDENCE 375/ 25 375/ 25 375/ 25 375/ 25 PORTION OF TAIL MISSING 0/ 0 0/ 0 0/ 0 11/ 1 SPARSE HAIR COAT: TOTAL 1/ 1 0/ 0 0/ 0 4/ 2 UNDERSIDE 0/ 0 0/ 0 0/ 0 3/ 1 LIMB(S) 1/ 1 0/ 0 0/ 0 1/ 1 NECK 0/ 0 0/ 0 0/ 0 1/ 1 NO GROSS LESIONS WERE IDENTIFIED AT NECROPSY ------------------------------------------------------------------------------------------------------------------------------------ MAXIMUM POSSIBLE INCIDENCE = (DAYS x RATS)/NUMBER OF RATS EXAMINED PER GROUP N/N = TOTAL NUMBER OF OBSERVATIONS/NUMBER OF RATS WITH OBSERVATION a. Dose administration occurred on Days 7 through 17 of gestation.



Table 2 Maternal Body Weights - Summary


Revised Draft Report Testing Facility Study No. 20024504 PROTOCOL 20024504: AN EMBRYO-FETAL DEVELOPMENT STUDY OF THYMOSIN BETA 4 (FORMULATED AS RGN-532) BY INTRAVENOUS INJECTION IN RATS TABLE 2 (PAGE 1): MATERNAL BODY WEIGHTS - SUMMARY ------------------------------------------------------------------------------------------------------------------------------------ GROUP 1 2 3 4 TEST MATERIAL CONTROL ARTICLE RGN-352 RGN-352 RGN-352 DOSE (MG/KG/DAY)a 0 (CONTROL) 20 60 180 ------------------------------------------------------------------------------------------------------------------------------------ RATS TESTED N 25 25 25 25 PREGNANT N 25 25 25 23 MATERNAL BODY WEIGHT (G) DAY 0 MEAN±S.D. 242.2 ± 7.5 241.8 ± 7.6 241.9 ± 7.5 243.0 ± 7.5 DAY 7 MEAN±S.D. 285.4 ± 12.7 284.6 ± 12.4 286.0 ± 10.5 287.9 ± 11.8 DAY 8 MEAN±S.D. 287.4 ± 13.6 287.0 ± 14.0 288.2 ± 10.6 290.4 ± 12.1 DAY 9 MEAN±S.D. 292.1 ± 13.0 291.2 ± 12.9 291.7 ± 11.5 294.5 ± 13.2 DAY 10 MEAN±S.D. 297.8 ± 15.4 296.6 ± 14.7 298.0 ± 11.4 301.2 ± 13.4 DAY 11 MEAN±S.D. 304.5 ± 16.2 303.1 ± 16.8 304.9 ± 13.3 306.5 ± 13.6 DAY 12 MEAN±S.D. 309.0 ± 15.3 304.3 ± 15.1 309.5 ± 12.8 311.7 ± 13.5 DAY 13 MEAN±S.D. 313.8 ± 15.6 310.3 ± 16.5 315.2 ± 13.4 317.2 ± 14.8 DAY 14 MEAN±S.D. 319.5 ± 18.2 315.3 ± 16.7 320.4 ± 13.7 322.6 ± 14.7 DAY 15 MEAN±S.D. 326.1 ± 18.7 323.6 ± 18.2 328.2 ± 14.1 330.5 ± 15.1 DAY 16 MEAN±S.D. 337.5 ± 20.0 333.8 ± 20.4 339.1 ± 15.7 342.5 ± 15.3 DAY 17 MEAN±S.D. 351.6 ± 22.2 347.5 ± 21.6 350.4 ± 16.6 357.0 ± 16.5 DAY 18 MEAN±S.D. 365.8 ± 21.9 362.7 ± 23.1 365.7 ± 17.0 372.9 ± 17.2 DAY 19 MEAN±S.D. 380.2 ± 23.8 378.7 ± 25.0 383.0 ± 18.4 389.4 ± 18.5 DAY 20 MEAN±S.D. 398.8 ± 25.5 395.8 ± 28.0 400.2 ± 19.0 407.5 ± 19.8 DAY 21 MEAN±S.D. 428.4 ± 29.6 424.6 ± 32.0 429.1 ± 20.6 437.0 ± 22.6 ------------------------------------------------------------------------------------------------------------------------------------ ALL WEIGHTS WERE RECORDED IN GRAMS (G). DAY = DAY OF GESTATION a. Dose administration occurred on Days 7 through 17 of gestation.



Table 3 Maternal Body Weight Changes - Summary


Revised Draft Report Testing Facility Study No. 20024504 PROTOCOL 20024504: AN EMBRYO-FETAL DEVELOPMENT STUDY OF THYMOSIN BETA 4 (FORMULATED AS RGN-532) BY INTRAVENOUS INJECTION IN RATS TABLE 3 (PAGE 1): MATERNAL BODY WEIGHT CHANGES - SUMMARY ------------------------------------------------------------------------------------------------------------------------------------ GROUP 1 2 3 4 TEST MATERIAL CONTROL ARTICLE RGN-352 RGN-352 RGN-352 DOSE (MG/KG/DAY)a 0 (CONTROL) 20 60 180 ------------------------------------------------------------------------------------------------------------------------------------ RATS TESTED N 25 25 25 25 PREGNANT N 25 25 25 23 MATERNAL BODY WEIGHT CHANGE (G) DAYS 0 - 7 MEAN±S.D. +43.2 ± 8.3 +42.8 ± 7.6 +44.1 ± 7.8 +45.0 ± 6.0 DAYS 7 - 10 MEAN±S.D. +12.4 ± 7.2 +11.9 ± 4.7 +12.0 ± 3.9 +13.3 ± 4.9 DAYS 10 - 12 MEAN±S.D. +11.2 ± 2.9 +7.8 ± 6.7* +11.4 ± 4.1 +10.5 ± 4.5 DAYS 12 - 15 MEAN±S.D. +17.2 ± 5.9 +19.2 ± 8.0 +18.7 ± 5.4 +18.8 ± 5.0 DAYS 15 - 18 MEAN±S.D. +39.7 ± 6.5 +39.2 ± 10.0 +37.5 ± 6.9 +42.3 ± 7.2 DAYS 7 - 18 MEAN±S.D. +80.4 ± 12.7 +78.1 ± 15.4 +79.7 ± 11.2 +85.0 ± 10.1 DAYS 18 - 21 MEAN±S.D. +62.6 ± 10.2 +61.8 ± 12.4 +63.4 ± 7.7 +64.1 ± 9.4 DAYS 7 - 21 MEAN±S.D. +143.0± 20.8 +139.9± 25.0 +143.1± 16.4 +149.0± 16.2 DAYS 0 - 21 MEAN±S.D. +186.2± 25.2 +182.8± 28.6 +187.2± 19.0 +194.0± 19.2 ------------------------------------------------------------------------------------------------------------------------------------ ALL WEIGHTS WERE RECORDED IN GRAMS (G). DAYS = DAYS OF GESTATION a. Dose administration occurred on Days 7 through 17 of gestation. * Significantly different from the control group value (p≤0.05).



Table 4 Maternal Absolute Food Consumption Values (g/day) - Summary


Revised Draft Report Testing Facility Study No. 20024504 PROTOCOL 20024504: AN EMBRYO-FETAL DEVELOPMENT STUDY OF THYMOSIN BETA 4 (FORMULATED AS RGN-532) BY INTRAVENOUS INJECTION IN RATS TABLE 4 (PAGE 1): MATERNAL ABSOLUTE FOOD CONSUMPTION VALUES (G/DAY) - SUMMARY ------------------------------------------------------------------------------------------------------------------------------------ GROUP 1 2 3 4 TEST MATERIAL CONTROL ARTICLE RGN-352 RGN-352 RGN-352 DOSE (MG/KG/DAY)a 0 (CONTROL) 20 60 180 ------------------------------------------------------------------------------------------------------------------------------------ RATS TESTED N 25 25 25 25 PREGNANT N 25 25 25 23 MATERNAL FOOD CONSUMPTION (G/DAY) DAYS 0 - 7 MEAN±S.D. 21.7 ± 2.4 22.0 ± 2.2 22.0 ± 2.6 22.0 ± 1.4 DAYS 7 - 10 MEAN±S.D. 22.7 ± 2.2 22.5 ± 2.4 22.7 ± 2.3 22.9 ± 1.5 DAYS 10 - 12 MEAN±S.D. 24.4 ± 2.4 23.1 ± 2.7 24.6 ± 2.4 24.4 ± 2.6 DAYS 12 - 15 MEAN±S.D. 24.7 ± 3.1 24.0 ± 2.1 24.8 ± 2.3 24.8 ± 2.5 DAYS 15 - 18 MEAN±S.D. 27.0 ± 2.8 27.0 ± 3.0 27.5 ± 2.6 27.8 ± 2.1 [ 24]b DAYS 7 - 18 MEAN±S.D. 24.7 ± 2.4 24.2 ± 2.2 24.9 ± 1.9 25.0 ± 1.8 [ 24]b DAYS 18 - 21 MEAN±S.D. 28.3 ± 3.0 26.8 ± 3.9 28.8 ± 2.7 28.6 ± 2.4 DAYS 7 - 21 MEAN±S.D. 25.5 ± 2.5 24.8 ± 2.4 25.7 ± 1.9 25.8 ± 1.7 DAYS 0 - 21 MEAN±S.D. 24.2 ± 2.3 23.8 ± 2.2 24.5 ± 1.9 24.5 ± 1.4 ------------------------------------------------------------------------------------------------------------------------------------ ALL WEIGHTS WERE RECORDED IN GRAMS (G). DAYS = DAYS OF GESTATION [ ] = NUMBER OF VALUES AVERAGED a. Dose administration occurred on Days 7 through 17 of gestation. b. Excludes a value that was associated with spillage.



Table 5 Maternal Relative Food Consumption Values (g/kg/day) - Summary


Revised Draft Report Testing Facility Study No. 20024504 PROTOCOL 20024504: AN EMBRYO-FETAL DEVELOPMENT STUDY OF THYMOSIN BETA 4 (FORMULATED AS RGN-532) BY INTRAVENOUS INJECTION IN RATS TABLE 5 (PAGE 1): MATERNAL RELATIVE FOOD CONSUMPTION VALUES (G/KG/DAY) - SUMMARY ------------------------------------------------------------------------------------------------------------------------------------ GROUP 1 2 3 4 TEST MATERIAL CONTROL ARTICLE RGN-352 RGN-352 RGN-352 DOSE (MG/KG/DAY)a 0 (CONTROL) 20 60 180 ------------------------------------------------------------------------------------------------------------------------------------ RATS TESTED N 25 25 25 25 PREGNANT N 25 25 25 23 MATERNAL FOOD CONSUMPTION (G/KG/DAY) DAYS 0 - 7 MEAN±S.D. 82.3 ± 7.0 83.3 ± 6.6 83.2 ± 8.9 83.0 ± 4.5 DAYS 7 - 10 MEAN±S.D. 77.9 ± 5.2 77.6 ± 6.8 77.9 ± 6.0 78.0 ± 4.1 DAYS 10 - 12 MEAN±S.D. 80.4 ± 5.9 76.5 ± 7.4 80.7 ± 6.2 79.6 ± 6.8 DAYS 12 - 15 MEAN±S.D. 77.8 ± 6.7 76.7 ± 5.0 77.7 ± 5.4 77.2 ± 6.6 DAYS 15 - 18 MEAN±S.D. 78.2 ± 6.4 78.9 ± 6.4 79.4 ± 5.6 79.2 ± 5.1 [ 24]b DAYS 7 - 18 MEAN±S.D. 78.2 ± 4.9 77.3 ± 5.0 78.6 ± 3.9 78.2 ± 4.4 [ 24]b DAYS 18 - 21 MEAN±S.D. 71.9 ± 5.1 68.7 ± 8.3 73.2 ± 6.5 71.3 ± 4.8 DAYS 7 - 21 MEAN±S.D. 76.4 ± 4.6 74.9 ± 5.4 77.0 ± 3.6 76.3 ± 3.7 DAYS 0 - 21 MEAN±S.D. 74.0 ± 4.2 73.2 ± 4.4 74.6 ± 4.0 73.9 ± 2.9 ------------------------------------------------------------------------------------------------------------------------------------ ALL WEIGHTS WERE RECORDED IN GRAMS (G). DAYS = DAYS OF GESTATION [ ] = NUMBER OF VALUES AVERAGED a. Dose administration occurred on Days 7 through 17 of gestation. b. Excludes a value that was associated with spillage.



Table 6 Caesarean-Sectioning Observations - Summary


Revised Draft Report Testing Facility Study No. 20024504 PROTOCOL 20024504: AN EMBRYO-FETAL DEVELOPMENT STUDY OF THYMOSIN BETA 4 (FORMULATED AS RGN-532) BY INTRAVENOUS INJECTION IN RATS TABLE 6 (PAGE 1): CAESAREAN-SECTIONING OBSERVATIONS - SUMMARY ------------------------------------------------------------------------------------------------------------------------------------ GROUP 1 2 3 4 TEST MATERIAL CONTROL ARTICLE RGN-352 RGN-352 RGN-352 DOSE (MG/KG/DAY)a 0 (CONTROL) 20 60 180 ------------------------------------------------------------------------------------------------------------------------------------ RATS TESTED N 25 25 25 25 PREGNANT N(%) 25(100.0) 25(100.0) 25(100.0) 23( 92.0) RATS PREGNANT AND CAESAREAN-SECTIONED ON DAY 21 OF GESTATION N 25 25 25 23 CORPORA LUTEA MEAN±S.D. 14.7 ± 2.0 15.2 ± 2.2 14.6 ± 1.9 14.9 ± 1.8 IMPLANTATIONS MEAN±S.D. 14.4 ± 1.9 14.3 ± 1.8 14.1 ± 1.9 14.6 ± 1.8 % PREIMPLANTATION LOSS MEAN±S.D. 1.5 ± 3.2 4.9 ± 9.1 3.0 ± 5.0 2.0 ± 3.2 LITTER SIZES MEAN±S.D. 13.3 ± 1.7 13.6 ± 2.5 13.5 ± 1.8 14.1 ± 1.7 LIVE FETUSES N 333 340 338 324 MEAN±S.D. 13.3 ± 1.7 13.6 ± 2.5 13.5 ± 1.8 14.1 ± 1.7 DEAD FETUSES N 0 0 0 0 RESORPTIONS MEAN±S.D. 1.1 ± 1.4 0.7 ± 1.4 0.6 ± 0.6 0.5 ± 0.8 EARLY RESORPTIONS N 27 18 15 11 MEAN±S.D. 1.1 ± 1.4 0.7 ± 1.4 0.6 ± 0.6 0.5 ± 0.8 LATE RESORPTIONS N 1 0 0 0 MEAN±S.D. 0.0 ± 0.2 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0 ------------------------------------------------------------------------------------------------------------------------------------ % PREIMPLANTATION LOSS = [(NUMBER OF CORPORA LUTEA - NUMBER OF IMPLANTATIONS) / NUMBER OF CORPORA LUTEA] x 100 a. Dose administration occurred on Days 7 through 17 of gestation.


Revised Draft Report Testing Facility Study No. 20024504 PROTOCOL 20024504: AN EMBRYO-FETAL DEVELOPMENT STUDY OF THYMOSIN BETA 4 (FORMULATED AS RGN-532) BY INTRAVENOUS INJECTION IN RATS TABLE 6 (PAGE 2): CAESAREAN-SECTIONING OBSERVATIONS - SUMMARY ------------------------------------------------------------------------------------------------------------------------------------ GROUP 1 2 3 4 TEST MATERIAL CONTROL ARTICLE RGN-352 RGN-352 RGN-352 DOSE (MG/KG/DAY)a 0 (CONTROL) 20 60 180 ------------------------------------------------------------------------------------------------------------------------------------ RATS TESTED N 25 25 25 25 PREGNANT N(%) 25(100.0) 25(100.0) 25(100.0) 23( 92.0) RATS PREGNANT AND CAESAREAN-SECTIONED ON DAY 21 OF GESTATION N 25 25 25 23 % POSTIMPLANTATION LOSS MEAN±S.D. 7.3 ± 9.2 5.2 ± 10.8 4.1 ± 4.4 3.1 ± 4.9 DAMS WITH ANY RESORPTIONS N(%) 12( 48.0) 8( 32.0) 13( 52.0) 8( 34.8) DAMS WITH ALL CONCEPTUSES RESORBED N(%) 0( 0.0) 0( 0.0) 0( 0.0) 0( 0.0) DAMS WITH VIABLE FETUSES N(%) 25(100.0) 25(100.0) 25(100.0) 23(100.0) PLACENTAE APPEARED NORMAL N(%) 25(100.0) 25(100.0) 25(100.0) 23(100.0) ------------------------------------------------------------------------------------------------------------------------------------ % POSTIMPLANTATION LOSS = [(NUMBER OF IMPLANTATIONS - NUMBER OF LIVE FETUSES)/NUMBER OF IMPLANTATIONS] X 100 a. Dose administration occurred on Days 7 through 17 of gestation.



Table 7 Litter Observations (Caesarean-Delivered Fetuses) - Summary


Revised Draft Report Testing Facility Study No. 20024504 PROTOCOL 20024504: AN EMBRYO-FETAL DEVELOPMENT STUDY OF THYMOSIN BETA 4 (FORMULATED AS RGN-532) BY INTRAVENOUS INJECTION IN RATS TABLE 7 (PAGE 1): LITTER OBSERVATIONS (CAESAREAN-DELIVERED FETUSES) - SUMMARY ------------------------------------------------------------------------------------------------------------------------------------ GROUP 1 2 3 4 TEST MATERIAL CONTROL ARTICLE RGN-352 RGN-352 RGN-352 DOSE (MG/KG/DAY)a 0 (CONTROL) 20 60 180 ------------------------------------------------------------------------------------------------------------------------------------ LITTERS WITH ONE OR MORE LIVE FETUSES N 25 25 25 23 IMPLANTATIONS MEAN±S.D. 14.4 ± 1.9 14.3 ± 1.8 14.1 ± 1.9 14.6 ± 1.8 LIVE FETUSES N 333 340 338 324 MEAN±S.D. 13.3 ± 1.7 13.6 ± 2.5 13.5 ± 1.8 14.1 ± 1.7 LIVE MALE FETUSES N 155 159 177 166 % LIVE MALE FETUSES/LITTER MEAN±S.D. 46.1 ± 14.3 46.0 ± 11.9 52.7 ± 17.4 51.4 ± 12.8 LIVE FETAL BODY WEIGHTS (GRAMS)/LITTER MEAN±S.D. 5.54 ± 0.32 5.59 ± 0.29 5.79 ± 0.28** 5.71 ± 0.31 MALE FETUSES MEAN±S.D. 5.67 ± 0.36 5.72 ± 0.38 5.92 ± 0.30* 5.84 ± 0.32 FEMALE FETUSES MEAN±S.D. 5.42 ± 0.33 5.49 ± 0.29 5.64 ± 0.28 5.54 ± 0.32 % RESORBED CONCEPTUSES/LITTER MEAN±S.D. 7.3 ± 9.2 5.3 ± 10.8 4.1 ± 4.4 3.1 ± 4.9 ------------------------------------------------------------------------------------------------------------------------------------ a. Dose administration occurred on Days 7 through 17 of gestation. * Significantly different from the control group value (p≤0.05). ** Significantly different from the control group value (p≤0.01).



Table 8 Fetal Alterations - Caesarean-Delivered Live Fetuses (Day 21 of Gestation) - Summary


Revised Draft Report Testing Facility Study No. 20024504 PROTOCOL 20024504: AN EMBRYO-FETAL DEVELOPMENT STUDY OF THYMOSIN BETA 4 (FORMULATED AS RGN-532) BY INTRAVENOUS INJECTION IN RATS TABLE 8 (PAGE 1): FETAL ALTERATIONS - CAESAREAN-DELIVERED LIVE FETUSES (DAY 21 OF GESTATION) - SUMMARY ------------------------------------------------------------------------------------------------------------------------------------ GROUP 1 2 3 4 TEST MATERIAL CONTROL ARTICLE RGN-352 RGN-352 RGN-352 DOSE (MG/KG/DAY)a 0 (CONTROL) 20 60 180 ------------------------------------------------------------------------------------------------------------------------------------ LITTERS EVALUATED N 25 25 25 23 LITTERS WITH LIVE FETUS(ES) N 25 25 25 23 FETUSES EVALUATED N 333 340 338 324 ------------------------------------------------------------------------------------------------------------------------------------ LITTERS WITH FETUSES WITH ANY ALTERATION OBSERVED N(%) 7( 28.0) 10( 40.0) 4( 16.0) 3( 13.0) FETUSES WITH ANY ALTERATION OBSERVED N(%) 8( 2.4) 15( 4.4)** 4( 1.2) 6( 1.8) % FETUSES WITH ANY ALTERATION/LITTER MEAN±S.D. 2.7 ± 4.8 4.6 ± 6.4 1.1 ± 2.7 1.8 ± 6.1 ------------------------------------------------------------------------------------------------------------------------------------ a. Dose administration occurred on Days 7 through 17 of gestation. ** Significantly different from the control group value (p≤0.01).



Table 9 Fetal Gross External Alterations - Caesarean-Delivered Live Fetuses

(Day 21 of Gestation) - Summary


Revised Draft Report Testing Facility Study No. 20024504 PROTOCOL 20024504: AN EMBRYO-FETAL DEVELOPMENT STUDY OF THYMOSIN BETA 4 (FORMULATED AS RGN-532) BY INTRAVENOUS INJECTION IN RATS TABLE 9 (PAGE 1): FETAL GROSS EXTERNAL ALTERATIONS - CAESAREAN-DELIVERED LIVE FETUSES (DAY 21 OF GESTATION) - SUMMARY ------------------------------------------------------------------------------------------------------------------------------------ GROUP 1 2 3 4 TEST MATERIAL CONTROL ARTICLE RGN-352 RGN-352 RGN-352 DOSE (MG/KG/DAY)a 0 (CONTROL) 20 60 180 ------------------------------------------------------------------------------------------------------------------------------------ LITTERS EVALUATED N 25 25 25 23 LITTERS WITH LIVE FETUS(ES) N 25 25 25 23 FETUSES EVALUATED N 333 340 338 324 LIVE N 333 340 338 324 ------------------------------------------------------------------------------------------------------------------------------------ NO FETAL ALTERATIONS WERE IDENTIFIED AT GROSS EXTERNAL EXAMINATION ------------------------------------------------------------------------------------------------------------------------------------ a. Dose administration occurred on Days 7 through 17 of gestation.



Table 10 Fetal Soft Tissue Alterations - Caesaraean-Delivered Live Fetuses (Day 21 of Gestation) -

Summary


Revised Draft Report Testing Facility Study No. 20024504 PROTOCOL 20024504: AN EMBRYO-FETAL DEVELOPMENT STUDY OF THYMOSIN BETA 4 (FORMULATED AS RGN-532) BY INTRAVENOUS INJECTION IN RATS TABLE 10 (PAGE 1): FETAL SOFT TISSUE ALTERATIONS - CAESAREAN-DELIVERED LIVE FETUSES (DAY 21 OF GESTATION) - SUMMARY ------------------------------------------------------------------------------------------------------------------------------------ GROUP 1 2 3 4 TEST MATERIAL CONTROL ARTICLE RGN-352 RGN-352 RGN-352 DOSE (MG/KG/DAY)a 0 (CONTROL) 20 60 180 ------------------------------------------------------------------------------------------------------------------------------------ LITTERS EVALUATED N 25 25 25 23 LITTERS WITH LIVE FETUS(ES) N 25 25 25 23 FETUSES EVALUATED N 161 164 161 159 LIVE N 161 164 161 159 ------------------------------------------------------------------------------------------------------------------------------------ VESSELS: AORTA PASSES DORSAL TO THE TRACHEA AND ESOPHAGUS LITTER INCIDENCE N(%) 1( 4.0) 0( 0.0) 0( 0.0) 0( 0.0) FETAL INCIDENCE N(%) 1( 0.6)b 0( 0.0) 0( 0.0) 0( 0.0) VESSELS: INNOMINATE ARTERY ABSENT LITTER INCIDENCE N(%) 1( 4.0) 1( 4.0) 0( 0.0) 0( 0.0) FETAL INCIDENCE N(%) 1( 0.6)b 1( 0.6) 0( 0.0) 0( 0.0) VESSELS: ARISE IN INCORRECT ORDER LITTER INCIDENCE N(%) 1( 4.0) 0( 0.0) 0( 0.0) 0( 0.0) FETAL INCIDENCE N(%) 1( 0.6)b 0( 0.0) 0( 0.0) 0( 0.0) ------------------------------------------------------------------------------------------------------------------------------------ a. Dose administration occurred on Days 7 through 17 of gestation. b. Fetus 4010-13 had other soft tissue alterations.



Table 11 Fetal Skeletal Alterations - Caesarean-Delivered Live Fetuses (Day 21 of Gestation) -

Summary


Revised Draft Report Testing Facility Study No. 20024504 PROTOCOL 20024504: AN EMBRYO-FETAL DEVELOPMENT STUDY OF THYMOSIN BETA 4 (FORMULATED AS RGN-532) BY INTRAVENOUS INJECTION IN RATS TABLE 11 (PAGE 1): FETAL SKELETAL ALTERATIONS - CAESAREAN-DELIVERED LIVE FETUSES (DAY 21 OF GESTATION) - SUMMARY (See the last page of this table for footnotes.) ------------------------------------------------------------------------------------------------------------------------------------ GROUP 1 2 3 4 TEST MATERIAL CONTROL ARTICLE RGN-352 RGN-352 RGN-352 DOSE (MG/KG/DAY)a 0 (CONTROL) 20 60 180 ------------------------------------------------------------------------------------------------------------------------------------ LITTERS EVALUATED N 25 25 25 23 LITTERS WITH LIVE FETUS(ES) N 25 25 25 23 FETUSES EVALUATED N 173 176 177 165 LIVE N 173 176 177 165 ------------------------------------------------------------------------------------------------------------------------------------ SKULL: EYE SOCKET, SMALL LITTER INCIDENCE N(%) 0( 0.0) 1( 4.0) 0( 0.0) 0( 0.0) FETAL INCIDENCE N(%) 0( 0.0) 1( 0.6)e 0( 0.0) 0( 0.0) SKULL: NASAL - FRONTAL, SUTURE LARGE LITTER INCIDENCE N(%) 0( 0.0) 0( 0.0) 0( 0.0) 1( 4.3) FETAL INCIDENCE N(%) 0( 0.0) 0( 0.0) 0( 0.0) 1( 0.6)k CERVICAL VERTEBRAE: ARCH, INCOMPLETELY OSSIFIED LITTER INCIDENCE N(%) 0( 0.0) 1( 4.0) 0( 0.0) 0( 0.0) FETAL INCIDENCE N(%) 0( 0.0) 2( 1.1)e 0( 0.0) 0( 0.0) CERVICAL VERTEBRAE: ARCHES, FUSED LITTER INCIDENCE N(%) 0( 0.0) 1( 4.0) 0( 0.0) 0( 0.0) FETAL INCIDENCE N(%) 0( 0.0) 1( 0.6)e 0( 0.0) 0( 0.0) CERVICAL VERTEBRAE: CERVICAL RIB PRESENT AT 7TH CERVICAL VERTEBRAE LITTER INCIDENCE N(%) 1( 4.0) 1( 4.0) 1( 4.0) 1( 4.3) FETAL INCIDENCE N(%) 1( 0.6) 1( 0.6) 1( 0.6) 1( 0.6) CERVICAL VERTEBRAE: ARCH, 6TH HAS THE APPEARANCE LIKE THE 7TH LITTER INCIDENCE N(%) 2( 8.0) 0( 0.0) 0( 0.0) 0( 0.0) FETAL INCIDENCE N(%) 2( 1.2) 0( 0.0) 0( 0.0) 0( 0.0) THORACIC VERTEBRAE: CENTRUM, BIFID LITTER INCIDENCE N(%) 1( 4.0) 4( 16.0) 1( 4.0) 0( 0.0) FETAL INCIDENCE N(%) 1( 0.6) 4( 2.3)e 1( 0.6) 0( 0.0) THORACIC VERTEBRAE: CENTRUM, UNILATERAL OSSIFICATION LITTER INCIDENCE N(%) 0( 0.0) 1( 4.0) 0( 0.0) 0( 0.0) FETAL INCIDENCE N(%) 0( 0.0) 1( 0.6)e 0( 0.0) 0( 0.0) ------------------------------------------------------------------------------------------------------------------------------------


Revised Draft Report Testing Facility Study No. 20024504 PROTOCOL 20024504: AN EMBRYO-FETAL DEVELOPMENT STUDY OF THYMOSIN BETA 4 (FORMULATED AS RGN-532) BY INTRAVENOUS INJECTION IN RATS TABLE 11 (PAGE 2): FETAL SKELETAL ALTERATIONS - CAESAREAN-DELIVERED LIVE FETUSES (DAY 21 OF GESTATION) - SUMMARY (See the last page of this table for footnotes.) ------------------------------------------------------------------------------------------------------------------------------------ GROUP 1 2 3 4 TEST MATERIAL CONTROL ARTICLE RGN-352 RGN-352 RGN-352 DOSE (MG/KG/DAY)a 0 (CONTROL) 20 60 180 ------------------------------------------------------------------------------------------------------------------------------------ LITTERS EVALUATED N 25 25 25 23 LITTERS WITH LIVE FETUS(ES) N 25 25 25 23 FETUSES EVALUATED N 173 176 177 165 LIVE N 173 176 177 165 ------------------------------------------------------------------------------------------------------------------------------------ THORACIC VERTEBRAE: ARCH, SMALL LITTER INCIDENCE N(%) 0( 0.0) 1( 4.0) 0( 0.0) 0( 0.0) FETAL INCIDENCE N(%) 0( 0.0) 1( 0.6)e 0( 0.0) 0( 0.0) RIBS: WAVY LITTER INCIDENCE N(%) 1( 4.0) 3( 12.0) 1( 4.0) 1( 4.3) FETAL INCIDENCE N(%) 1( 0.6) 4( 2.3)b,c,f,g 1( 0.6)h 3( 1.8)i-k RIBS: INCOMPLETELY OSSIFIED LITTER INCIDENCE N(%) 2( 8.0) 4( 16.0) 1( 4.0) 2( 8.7) FETAL INCIDENCE N(%) 2( 1.2) 6( 3.4)b,c,f,g 1( 0.6)h 4( 2.4)i-k RIBS: FUSED LITTER INCIDENCE N(%) 0( 0.0) 1( 4.0) 0( 0.0) 0( 0.0) FETAL INCIDENCE N(%) 0( 0.0) 1( 0.6)e 0( 0.0) 0( 0.0) RIBS: SHORT LITTER INCIDENCE N(%) 0( 0.0) 0( 0.0) 0( 0.0) 1( 4.3) FETAL INCIDENCE N(%) 0( 0.0) 0( 0.0) 0( 0.0) 1( 0.6) RIBS: THICKENED LITTER INCIDENCE N(%) 0( 0.0) 0( 0.0) 0( 0.0) 1( 4.3) FETAL INCIDENCE N(%) 0( 0.0) 0( 0.0) 0( 0.0) 1( 0.6)k ------------------------------------------------------------------------------------------------------------------------------------


Final Report Testing Facility Study No. 20024504 PROTOCOL 20024504: AN EMBRYO-FETAL DEVELOPMENT STUDY OF THYMOSIN BETA 4 (FORMULATED AS RGN-532) BY INTRAVENOUS INJECTION IN RATS TABLE 11 (PAGE 3): FETAL SKELETAL ALTERATIONS - CAESAREAN-DELIVERED LIVE FETUSES (DAY 21 OF GESTATION) - SUMMARY ------------------------------------------------------------------------------------------------------------------------------------ GROUP 1 2 3 4 TEST MATERIAL CONTROL ARTICLE RGN-352 RGN-352 RGN-352 DOSE (MG/KG/DAY)a 0 (CONTROL) 20 60 180 ------------------------------------------------------------------------------------------------------------------------------------ LITTERS EVALUATED N 25 25 25 23 LITTERS WITH LIVE FETUS(ES) N 25 25 25 23 FETUSES EVALUATED N 173 176 177 165 LIVE N 173 176 177 165 ------------------------------------------------------------------------------------------------------------------------------------ STERNAL CENTRA: ASYMMETRIC LITTER INCIDENCE N(%) 0( 0.0) 1( 4.0) 0( 0.0) 0( 0.0) FETAL INCIDENCE N(%) 0( 0.0) 1( 0.6)e 0( 0.0) 0( 0.0) STERNAL CENTRA: DUPLICATED LITTER INCIDENCE N(%) 0( 0.0) 1( 4.0) 0( 0.0) 0( 0.0) FETAL INCIDENCE N(%) 0( 0.0) 1( 0.6)d 0( 0.0) 0( 0.0) STERNAL CENTRA: INCOMPLETELY OSSIFIED LITTER INCIDENCE N(%) 0( 0.0) 1( 4.0) 0( 0.0) 0( 0.0) FETAL INCIDENCE N(%) 0( 0.0) 1( 0.6) 0( 0.0) 0( 0.0) XIPHOID: DUPLICATED LITTER INCIDENCE N(%) 0( 0.0) 1( 4.0) 0( 0.0) 0( 0.0) FETAL INCIDENCE N(%) 0( 0.0) 1( 0.6)d 0( 0.0) 0( 0.0) PELVIS: PUBIS, INCOMPLETELY OSSIFIED LITTER INCIDENCE N(%) 0( 0.0) 0( 0.0) 1( 4.0) 0( 0.0) FETAL INCIDENCE N(%) 0( 0.0) 0( 0.0) 1( 0.6) 0( 0.0) ------------------------------------------------------------------------------------------------------------------------------------ a. Dose administration occurred on Days 7 through 17 of gestation. b. Fetus 4037-1 had other skeletal alterations. c. Fetus 4037-3 had other skeletal alterations. d. Fetus 4043-3 had other skeletal alterations. e. Fetus 4045-1 had other skeletal alterations. f. Fetus 4048-15 had other skeletal alterations. g. Fetus 4049-7 had other skeletal alterations. h. Fetus 4066-5 had other skeletal alterations. i. Fetus 4087-3 had other skeletal alterations. j. Fetus 4087-9 had other skeletal alterations. k. Fetus 4087-11 had other skeletal alterations.



Table 12 Fetal Ossification Sites - Caesarean-Delivered Live Fetuses (Day 21 of Gestation) -

Summary


Revised Draft Report Testing Facility Study No. 20024504 PROTOCOL 20024504: AN EMBRYO-FETAL DEVELOPMENT STUDY OF THYMOSIN BETA 4 (FORMULATED AS RGN-532) BY INTRAVENOUS INJECTION IN RATS TABLE 12 (PAGE 1): FETAL OSSIFICATION SITES - CAESAREAN-DELIVERED LIVE FETUSES (DAY 21 OF GESTATION) - SUMMARY ------------------------------------------------------------------------------------------------------------------------------------ GROUP 1 2 3 4 TEST MATERIAL CONTROL ARTICLE RGN-352 RGN-352 RGN-352 DOSE (MG/KG/DAY)a 0 (CONTROL) 20 60 180 ------------------------------------------------------------------------------------------------------------------------------------ LITTERS EXAMINED N 25 25 25 23 FETUSES EXAMINED N 173 176 177 165 ------------------------------------------------------------------------------------------------------------------------------------ OSSIFICATION SITES PER FETUS PER LITTER HYOID MEAN±S.D. 0.99 ± 0.04 1.00 ± 0.00 1.00 ± 0.00 1.00 ± 0.00 VERTEBRAE CERVICAL MEAN±S.D. 7.00 ± 0.00 7.00 ± 0.00 7.00 ± 0.00 7.00 ± 0.00 THORACIC MEAN±S.D. 13.09 ± 0.15 13.08 ± 0.10 13.09 ± 0.15 13.10 ± 0.18 LUMBAR MEAN±S.D. 5.90 ± 0.15 5.92 ± 0.10 5.90 ± 0.15 5.90 ± 0.18 SACRAL MEAN±S.D. 3.00 ± 0.00 3.00 ± 0.00 3.00 ± 0.00 3.00 ± 0.00 CAUDAL MEAN±S.D. 7.10 ± 0.68 7.34 ± 0.73 7.49 ± 0.64 7.39 ± 0.72 RIBS (PAIRS) MEAN±S.D. 13.06 ± 0.11 13.06 ± 0.08 13.08 ± 0.12 13.08 ± 0.15 STERNUM MANUBRIUM MEAN±S.D. 1.00 ± 0.00 1.00 ± 0.00 1.00 ± 0.00 1.00 ± 0.00 STERNAL CENTERS MEAN±S.D. 4.00 ± 0.00 3.98 ± 0.06 4.00 ± 0.00 4.00 ± 0.00 XIPHOID MEAN±S.D. 1.00 ± 0.00 1.00 ± 0.00 1.00 ± 0.00 1.00 ± 0.00 FORELIMB b CARPALS MEAN±S.D. 0.00 ± 0.00 0.00 ± 0.00 0.00 ± 0.00 0.00 ± 0.00 METACARPALS MEAN±S.D. 4.00 ± 0.00 4.00 ± 0.00 4.00 ± 0.00 4.00 ± 0.00 DIGITS MEAN±S.D. 5.00 ± 0.00 5.00 ± 0.00 5.00 ± 0.00 5.00 ± 0.00 PHALANGES MEAN±S.D. 7.54 ± 0.70 7.84 ± 0.72 8.03 ± 0.76 7.90 ± 0.82 HINDLIMB b TARSALS MEAN±S.D. 0.01 ± 0.06 0.03 ± 0.07 0.02 ± 0.06 0.01 ± 0.04 METATARSALS MEAN±S.D. 4.80 ± 0.20 4.84 ± 0.18 4.92 ± 0.16 4.85 ± 0.20 DIGITS MEAN±S.D. 5.00 ± 0.00 5.00 ± 0.00 5.00 ± 0.00 5.00 ± 0.00 PHALANGES MEAN±S.D. 5.73 ± 0.74 6.24 ± 0.98 6.54 ± 0.99** 6.30 ± 0.86* ------------------------------------------------------------------------------------------------------------------------------------ a. Dose administration occurred on Days 7 through 17 of gestation. b. Calculated as average per limb. * Significantly different from the control group value (p≤0.05). ** Significantly different from the control group value (p≤0.01).



Appendix 1 Protocol and Deviations


FINAL PROTOCOL


An Embryo-fetal Development Study of Thymosin beta 4 (formulated as RGN-352) by Intravenous Injection in Rats




Preclinical Services, Pennsylvania (PCS-PA) 905 Sheehy Drive, Building A


1 May 2012

Page 1 of 25



TABLE OF CONTENTS

1. OBJECTIVES ..........................................................................................................................3

2. PROPOSED STUDY SCHEDULE ........................................................................................3

3. GUIDELINES FOR STUDY DESIGN ...................................................................................3

4. REGULATORY COMPLIANCE ...........................................................................................3

5. QUALITY ASSURANCE .......................................................................................................4

6. SPONSOR ...............................................................................................................................5

7. RESPONSIBLE PERSONNEL...............................................................................................5

8. TEST AND CONTROL ARTICLES ......................................................................................6

9. SAFETY ..................................................................................................................................7

10. DOSE FORMULATION AND ANALYSIS ..........................................................................7

11. TEST SYSTEM .......................................................................................................................9

12. HUSBANDRY ......................................................................................................................10

13. EXPERIMENTAL DESIGN .................................................................................................12

14. IN-LIFE PROCEDURES, OBSERVATIONS, AND MEASUREMENTS .........................13

15. TERMINAL PROCEDURES ...............................................................................................14

16. FETAL EXAMINATIONS ...................................................................................................16

17. COMPUTERIZED SYSTEMS .............................................................................................17

18. STATISTICAL ANALYSIS .................................................................................................18

19. AMENDMENTS AND DEVIATIONS ................................................................................19

20. RETENTION OF RECORDS, SAMPLES, AND SPECIMENS .........................................19

21. REPORTING .........................................................................................................................20

22. ANIMAL WELFARE ...........................................................................................................21

23. REFERENCES ......................................................................................................................22

24. TESTING FACILITY APPROVAL .....................................................................................23

25. SPONSOR APPROVAL .......................................................................................................25



1. OBJECTIVES The objectives of this study are to detect adverse effects of thymosin beta 4 (Tβ4, formulated as RGN-352) on pregnant Crl:CD(SD) females and development of the embryo and fetus consequent to exposure of the female from implantation to closure of the hard palate. This study evaluates ICH Harmonised Tripartite Guideline stages C to D of the reproductive process.

2. PROPOSED STUDY SCHEDULE Proposed study dates are listed below. Actual applicable dates will be included in the Final Report.

Animal Arrival: 01 May 2012

Cohabitation: 07 May 2012 PM - 12 May 2012 AM

Transfer of Predose Formulation Samples: 15 May 2012

Initiation of Dosing: 15 May 2012 (Day 7 of presumed gestation)

Completion of Dosing: 29 May 2012 (Day 17 of presumed gestation)

Transfer of Last Dose Formulation Samples: 29 May 2012

Completion of In-life: 29 May 2012 - 02 Jun 2012 (Day 21 of presumed gestation)

Dose Formulation Draft Report: 10 Aug 2012

Audited Draft Report: 24 Aug 2012

3. GUIDELINES FOR STUDY DESIGN The design of this study was based on the study objective(s), the overall product development strategy for the test article, and the following study design guidelines:

• ICH Harmonised Tripartite Guideline M3 (R2). Nonclinical Safety Studies for the Conduct of Human Clinical Trials for Pharmaceuticals.

• ICH Harmonised Tripartite Guideline S5 (R2). Guideline for Industry: detection of toxicity to reproduction for medicinal products & toxicity to male fertility.

4. REGULATORY COMPLIANCE The study will be performed in accordance with the U.S. Department of Health and Human Services, Food and Drug Administration, United States Code of Federal Regulations, Title 21, Part 58: Good Laboratory Practice for Nonclinical Laboratory Studies and as accepted by Regulatory Authorities throughout the European Community (OECD Principles of Good Laboratory Practice) and Japan (MHLW).



Exceptions to GLPs include the following study elements:

• Characterization of the test and control articles was performed by a Sponsor subcontractor at a facility that follows FDA current Good Manufacturing Practice (cGMP) regulations.

5. QUALITY ASSURANCE

5.1. Testing Facility The Testing Facility Quality Assurance Unit (QAU) will monitor the study to assure the facilities, equipment, personnel, methods, practices, records, and controls are in conformance with Good Laboratory Practice regulations. The QAU will review the protocol, conduct inspections at intervals adequate to assure the integrity of the study, and audit the Final Report to assure that it accurately describes the methods and standard operating procedures and that the reported results accurately reflect the raw data of the study.

The Testing Facility QAU contact for this study is indicated below:

Karen J. Waetjen, M.S., RQAP-GLP Manager, Regulatory Compliance Preclinical Services Charles River 905 Sheehy Drive, Building A Horsham, PA 19044 Tel: 215.443.8710 Fax: 215.443.8587 E-mail: [email protected]



5.2. Testing Facility-designated Subcontractor The following study phase performed by Testing Facility-designated subcontractor will be audited by the Testing Facility QAU:

• Dose formulation analysis For all study phase(s) inspected by subcontractor QAU(s), copies of each periodic inspection report will be made available to the Study Director, Testing Facility Management, and the Testing Facility QAU.

6. SPONSOR Sponsor Representative and Study Monitor David Crockford, Vice President Clinical and Regulatory Affairs RegeneRx Biopharmaceuticals, Inc. Tel: 301.208.9191 Fax: 617.285.5588 E-mail: [email protected]

7. RESPONSIBLE PERSONNEL Study Director Valerie A. Sharper, MS Address as cited for Testing Facility Tel: 267.532.3838 Fax: 215.443.8587 E-mail: [email protected]

Management Contact Alan M. Hoberman, PhD, DABT, Fellow ATS Address as cited for Testing Facility Tel: 215.443.8710 Fax: 215.443.8587 E-mail: [email protected]

Principal Investigator (PI) Dose Formulation Analysis Jason Sarsoza, BSc

Scientist I Charles River Laboratories Preclinical Services 905 Sheehy Drive, Building A Horsham, PA 19044 Tel: 267.532.3771 Fax: 215.443.8587 E-mail: [email protected]



The PI is required to report any deviations or other circumstances that could affect the quality or integrity of the study to the Study Director in a timely manner. The PI will provide a report addressing their assigned phase of the study, which will be included as an appendix to the Final Report. Each phase report will include the following:

• A Statement of Compliance

• A QA Statement

• The archive site and storage duration for all records, samples, specimens and reports generated from the phase or segment

• A listing of critical computerized systems used in the conduct and/or interpretation of the assigned study phase

8. TEST AND CONTROL ARTICLES

8.1. Test Article Identification: Thymosin beta 4 (Tβ4) formulated as RGN-352 (Tβ4 Injectable

Solution)

Supplier: Sponsor

Batch (Lot) Number: 330-05-001

Expiration Date: To be documented in the raw data

Physical Description: colorless, clear liquid, 2.25 mL/vial



8.2. Control Article Identification: RGN-352 Placebo (Vehicle for RGN-352) Supplier: Sponsor

Batch (Lot) Number: 330-06-001

Expiration Date: To be documented in the raw data

Physical Description: colorless, clear liquid, 5.5 mL/vial


8.3. Test Material Shipment All test material shipments should be addressed to the attention of Julian Gulbinski ([email protected]), Senior Manager Laboratory Sciences, at the previously cited Testing Facility address.



Shipments should include information concerning storage conditions and shipping cartons should be labeled appropriately. The recipient should be notified in advance of shipment.

8.4. Test and Control Article Characterization The Sponsor will provide to the Testing Facility documentation of the identity, strength, purity, and composition for the test and control articles. A Certificate of Analysis or equivalent documentation will be provided for inclusion in the Final Report. The Sponsor will also provide information concerning the regulatory standard that was followed for these evaluations.

The Sponsor has appropriate documentation on file concerning the method of synthesis of the active ingredient, Tβ4, and fabrication or derivation of the test and control articles, and this information is available to the appropriate regulatory agencies should it be requested.

8.5. Analysis of Test Article Stability of the formulated test article for the duration of the study will be confirmed by comparison of the results of the concentration analysis conducted for samples collected before the first dose administration and samples collected on the last day of administration.

8.6. Reserve Samples For each batch (lot) of test and control article(s), a reserve sample (1 vial) will be collected and maintained under the appropriate storage conditions by the Testing Facility, pursuant to 21 CFR 58.190.

8.7. Test and Control Article Inventory and Disposition Records of the receipt, distribution, storage, and disposition of test and control articles (including empty containers) will be maintained. With the exception of reserve samples, all unused Sponsor-supplied bulk test and control articles will be retained for use in future studies with this test article or returned to the Sponsor before issue of the final report. All empty containers will be maintained for the duration of the study.

9. SAFETY The following safety instructions apply to this study:

Gloves, dust-mist/HEPA-filtered mask, appropriate eye protection and uniform/lab coat.

The Material Safety Data Sheet (MSDS) will be maintained with the raw data.

10. DOSE FORMULATION AND ANALYSIS

10.1. Preparation of Control Article The control article, RGN-352 Placebo, will be administered as received. Upon receipt, the control article will be dispensed daily for administration to Group 1 control animals. The control article will be removed from the refrigerator at least 30 minutes before dosing to allow it to come to room temperature.



Any residual volumes from partially used vials will be discarded before the Final Report is issued.

10.2. Preparation of Test Article The test article, formulated as RGN-352, will be administered as received. Upon receipt, the test article will be dispensed daily for administration to Groups 2, 3 and 4. The prepared test article will be removed from the refrigerator at least 30 minutes before dosing to allow it to come to room temperature.

Any residual volumes from partially used vials will be discarded before the Final Report is issued.

10.3. Sample Collection and Analysis Dose formulation samples will be collected for analysis as indicated in the following table. Additional samples may be collected and analyzed at the discretion of the Study Director.

Dose Formulation Sample Collection Schedule

Interval Concentration Homogeneity Stability Before First Administrationa All groups N/A N/A Last Day of Administration All groups N/A N/A

N/A = Not applicable. a. Results of the concentration analysis of the last day of administration samples from the most recently previously

performed study (Testing Facility Study No. 20024503) were within the acceptance criteria and are within 30 days from the first administration on this study (Testing Facility Study No. 20024504), analysis of the predose samples will not be required.

Samples to be analyzed will be submitted on the date prepared. All samples to be analyzed will be transferred (ambient conditions) to the analytical laboratory at the Testing Facility. Upon receipt at the analytical laboratory, the samples will be stored refrigerated (2°C to 8°C) until analysis. Any residual/retained analytical samples (and test article used in analysis) will be discarded before the Final Report is issued.

10.3.1. Analytical Method Analyses described below will be performed by HPLC-UV using validated analytical procedure TYMN03 - Analytical Procedure for the Analysis of Thymosin Beta 4 in RGN-352 Placebo by HPLC-UV; validated under PCS-PA Protocol No. 20019481.

10.3.1.1. Concentration Analysis Samples for Analysis: Single sample from one vial; transferred to Charles River’s

Analytical Chemistry Laboratory.

Backup Samples: Single sample from one vial; maintained at the Testing Facility. Backup samples may be analyzed at the discretion of the Study Director.



Sample Volume: 2.25 mL


Acceptance Criteria: The criteria for acceptability will be mean sample concentration results within or equal to ± 10% of theoretical concentration. Each individual sample concentration result within or equal to ± 15%.

10.3.1.2. Homogeneity Analysis The test article Tβ4, formulated and vialed as RGN-352 (100 mg/mL), yields a sterile aqueous, homogeneous solution. Therefore, homogeneity analyses of the test article will not be conducted. The test article will not be mixed with the placebo (RGN-352 excipients minus Tβ4) in this study.

10.3.1.3. Stability Analysis Stability of the formulated test article for the duration of the study will be confirmed by comparison of the results of the concentration analysis conducted for samples collected before the first dose administration and samples collected on the last day of administration.

11. TEST SYSTEM Species: Rat

Strain: Crl:CD(SD) Sprague Dawley

Condition: Virgin

Source: Charles River Laboratories, Kingston, NY

Number of Females Ordered: 120

Number of Females Assigned To Study: 100

Age/Weight: Rats will be ordered to have body weights of 201 g to 225 g each at receipt, at which time they will be expected to be approximately 60 days of age.

The actual body weight will be recorded the day after receipt. The actual age, weight, and number of animals received will be listed in the Final Report.

11.1. Justification of Test System and Number of Animals The test system was selected because: 1) it is a standard species accepted for use in embryo-fetal development studies; 2) this species and strain has been demonstrated to be sensitive to developmental toxicants; and 3) historical data and experience exist at the Testing Facility.

The number of animals chosen for this study is the smallest number considered necessary to provide the minimum number of pregnancies recommended by the applicable guidelines.



11.2. Animal Identification Female rats are permanently identified using Monel® self-piercing ear tags. Male rats are permanently identified by tattoo and given permanent unique identification numbers upon assignment to the Testing Facility's breeder male rat population. Female rats are assigned temporary numbers at receipt and given unique permanent identification numbers when assigned to the study on the basis of Day 0 of presumed gestation body weights.

11.3. Environmental Acclimation After receipt at the Testing Facility, the rats will be acclimated for at least 5 days prior to cohabitation.

11.4. Insemination/Mating Female rats will be mated with breeder male rats of the same source and strain, 1 male per female rat. The cohabitation period will consist of a maximum of 5 days. Female rats with spermatozoa observed in a smear of the vaginal contents and/or a copulatory plug observed in situ will be considered to be Day 0 of presumed gestation.

11.5. Selection, Assignment, and Replacement of Animals Healthy mated females will be assigned to groups using a computer-based weight-ordered randomization procedure, based on Day 0 of presumed gestation body weights. Assigned animals that die or are euthanized for reasons considered unrelated to toxicity will be replaced if possible, at the discretion of the Study Director, with spare animals. Any animal replacement following randomization will be documented in the study file and presented in the final report.

11.6. Animal Disposition The disposition of all animals will be documented in the study records. Any remaining animals not selected for study will be assigned to the Testing Facility’s general population or euthanized.

12. HUSBANDRY

12.1. Housing All cage sizes and housing conditions are in compliance with the Guide for the Care and Use of Laboratory Animals1

The rats will be individually housed in stainless steel, wire-bottomed cages, except during the cohabitation period. During cohabitation, each pair of rats will be housed in the male rat's nesting box. No nesting materials will be supplied to female rats that are individually housed following completion of the cohabitation period because the female rats will be euthanized before parturition is expected. Female rats with no confirmed date of mating will be assigned to the Testing Facility’s general population or euthanized.

.



12.2. Environmental Conditions The targeted conditions for animal room environment will be as follows:

Temperature: Room temperature will be maintained at 64°F to 79°F (18°C to 26°C) and monitored constantly.

Humidity: Room humidity will be monitored constantly and maintained at 30% to 70%.

Ventilation: The animal room is independently supplied with at least 10 changes per hour of 100% fresh air that has been passed through 99.97% HEPA filters.

Light Cycle: An automatically controlled 12-hour light:12-hour dark fluorescent light cycle will be maintained. Each dark period will begin at 1900 hours (± 30 minutes). The light cycle may be adjusted by the Study Director or designee if deemed necessary to accommodate scheduled laboratory activities. Any such adjustment will be documented in the raw data.

12.3. Food Rats will be given Certified Rodent Diet® #5002 (PMI® Nutrition International) available ad libitum from individual feeders. The Study Director is not aware of any potential contaminants likely to be present in the certified diet that would interfere with the results of this study. Therefore, no analyses other than those routinely performed by the feed supplier or those mentioned in this protocol will be conducted.

12.4. Water Water will be available ad libitum from individual bottles attached to the cages and/or from an automatic watering access system. All water will be from a local source and passed through a reverse osmosis membrane before use. Chlorine will be added to the processed water as a bacteriostat; processed water is expected to contain no more than 1.2 ppm chlorine at the time of analysis. Water is analyzed monthly for possible bacterial contamination and twice annually for possible chemical contamination. The Study Director is not aware of any potential contaminants likely to be present in the drinking water at levels that would interfere with the results of this study. Therefore, no analyses other than those mentioned in this protocol will be conducted.

12.5. Animal Enrichment Chewable Nylabones® will be supplied to all rats during the course of the study. Analyses for possible contamination are conducted on each lot of Nylabones® and documented in the raw data.



The Study Director is not aware of any potential contaminants likely to be present in the chewable enrichment devices at levels that would interfere with the results of this study. Therefore, no analyses other than those routinely performed by the enrichment device suppliers or those mentioned in this protocol will be conducted.

12.6. Veterinary Care Veterinary care will be available throughout the course of the study, and animals will be examined by the veterinary staff as warranted by clinical signs or other changes. All veterinary examinations and recommended therapeutic treatments, if any, will be documented in the study records.

13. EXPERIMENTAL DESIGN

Experimental Design


Dose Level (mg/kg/day)


Dose Volume (mL/kg) No. of Animals

1 Control Article 0 0 1.8 25 2

RGN-352 20 100 0.2 25

3 60 100 0.6 25 4 180 100 1.8 25

13.1. Administration of Test and Control Articles Female rats will be administered the test article and/or the control article by once daily intravenous bolus injection via the lateral tail vein on Days 7 through 17 of presumed gestation. Doses will be adjusted based on the most recently recorded body weight and administered at approximately the same time each day.

13.2. Justification of Route and Dose Levels The intravenous route was selected for use because it is one of the proposed systemic routes for clinical use.

Dose level selections for the test article in this study essentially bracket the dose level selections planned for RGN-352 in future clinical trials. The 180 mg/kg dose level is 6 times higher than the highest dose anticipated for administration in man. The low dose level of 20 mg/kg is 3.32 times higher than the lowest dose anticipated for administration in man. If an NOAEL cannot be established for the mated female and the development of the embryo and fetus from implantation to closure of the hard palate, then at least a 6-fold margin of safety would be established based on planned dose levels for RGN-352 in man.



Dosages were also selected on the basis of a dosage-range developmental toxicity study (Testing Facility Study No. 20024503). In that study, RGN-352 was administered once daily by intravenous bolus injection from gestation Days 7 to 17 at levels of 10, 30, 100 and 300 mg/kg/day. There were no test article-related adverse clinical observations and body weight gains and feed consumption values were comparable among the five dose groups for the entire dosage period. All Caesarean-sectioning and litter parameters were comparable among the five dose groups and there were no fetal gross external alterations.

14. IN-LIFE PROCEDURES, OBSERVATIONS, AND MEASUREMENTS The in-life procedures, observations, and measurements listed below will be performed for all rats.

14.1. Viability Checks Frequency: At least twice daily.

Procedure: Viability will be recorded.

14.2. Clinical Observations Clinical observations may be recorded more frequently than cited in the following sections.

14.2.1. General Appearance Frequency: At least weekly during the acclimation period, daily before

administration during the dosing period, and once daily during the postdose period.

Procedure: Clinical observations will be recorded.

14.2.2. Postdose Observations Frequency: Postdose observations will be recorded between 1 and 2 hours after

dose administration. Time intervals for postdose observations may be adjusted if deemed appropriate by the Study Director or designee during the course of the study. Such adjustments will be documented in the raw data.

Procedure: Clinical observations will be recorded.

14.3. Body Weights Frequency: At least weekly during the acclimation period, on Day 0 of

presumed gestation, and daily during the dosing and postdose periods.

Procedure: Body weights will be recorded.



14.4. Food Consumption Frequency: Days 0, 7, 10, 12, 15, 18 and 21 of presumed gestation

Procedure: Weights of feed left and feed fed will be recorded.

15. TERMINAL PROCEDURES Terminal procedures are summarized in the following table:

Terminal Procedures for Study Animals

Group No.

No. of Animals

Scheduled Euthanasia

Day

Necropsy Procedures

Histology Histopathology

Ovarian/ Uterine

Examination Necropsy Tissue

Collection Organ

Weights 1 25

21 Full Exam X X -

- - 2 25 - -

3 25 - -

4 25 - - Unscheduled Deaths Full Exam X X - - -

X = Procedure to be conducted; - = Not applicable.

15.1. Method of Euthanasia Rats will be euthanized by carbon dioxide asphyxiation. Live fetuses will be euthanized by an intraperitoneal injection of sodium pentobarbital.

15.2. Unscheduled Deaths Rats that die or are euthanized before scheduled termination will be examined for the cause of death or condition as soon as possible after the observation is made. The rats will be examined for gross lesions. See Section 15.5. (Tissue Collection and Preservation) for additional tissues that will be retained for possible further evaluation. Additional tissues may be retained at the discretion of the Study Director.

Pregnancy status and uterine contents will be recorded, and aborted fetuses, conceptuses in utero and/or delivered pups will be examined to the extent possible, using the same methods described for term fetuses.

Uteri of apparently nonpregnant rats will be examined while being pressed between glass plates to confirm the absence of implantation sites. Uteri and ovaries of apparently nonpregnant rats will be retained in neutral buffered 10% formalin and may be discarded when authorized by the Study Director.

15.3. Scheduled Euthanasia On Day 21 of presumed gestation, female rats will be Caesarean-sectioned and examined for gross lesions as further described in Sections 15.3.1 (Ovarian and Uterine Examination) and 15.4 (Necropsy).



15.3.1. Ovarian and Uterine Examinations The reproductive tract will be dissected from the abdominal cavity. The uterus will be opened, and the contents will be examined. The fetuses will be removed from the uterus and placed in individual containers (or a tray).

The ovaries and uterus will be examined for number and distribution of:

• Corpora Lutea

• Implantation Sites

• Placentae (size, color or shape) - any abnormalities will be recorded

• Live and Dead Fetuses

• Early and Late Resorptions Uteri of apparently nonpregnant rats will be examined while being pressed between glass plates to confirm the absence of implantation sites. Uteri and ovaries of apparently nonpregnant rats will be retained in 10% neutral buffered formalin and may be discarded when authorized by the Study Director.

15.4. Necropsy A gross necropsy of the thoracic, abdominal and pelvic viscera will be performed at scheduled euthanasia for all rats. Unless specifically cited in Section 15.5 (Tissue Collection and Preservation), all tissues will be discarded.

Images will be generated for illustration of or consultation on gross observations. Generation of such images will be documented. Images and associated documentation will be retained and archived.

15.5. Tissue Collection and Preservation Representative samples of the tissues identified in the Tissue Collection and Preservation table will be collected from all rats that are found dead or euthanized before scheduled termination and all rats that survive to scheduled euthanasia and will be preserved in 10% neutral buffered formalin, unless otherwise indicated. Additional tissues may be collected at the discretion of the Study Director to elucidate abnormal findings. Unless specifically cited below, all other tissues will be discarded.

A table of random units will be used to select one control group rat from which all tissues examined at necropsy will be retained, in order to provide control tissues for any possible histopathological evaluations of gross lesions.



Tissue Collection and Preservation

Tissue Collect Comment Animal identification X All rats with gross lesions and/or retained tissues.

Cervix X Collect with the uterus. All nonpregnant rats. Salivary glands X Rats with excess salivation (ptyalism) at any time during the study.

Gross lesions/masses X All rats. Heart X Rats found dead or euthanized before scheduled termination.

Kidney X Rats found dead or euthanized before scheduled termination. Liver X Rats found dead or euthanized before scheduled termination. Lung X Rats found dead or euthanized before scheduled termination.

Ovaries X All nonpregnant rats. Spleen X Rats found dead or euthanized before scheduled termination.

Stomach X Rats found dead or euthanized before scheduled termination. Uterus X All nonpregnant rats.

X = Procedure to be conducted

16. FETAL EXAMINATIONS Representative photographs of external, visceral and skeletal abnormalities will be taken. Photographs will not be included in the report, but will be retained as electronic images and archived with the raw data.

16.1. External Abnormalities and Sex Fetuses will be examined for sex and for external abnormalities. Late resorptions and dead fetuses will be examined for sex and for external abnormalities to the extent possible.

16.2. Body Weights and Identification The body weight of each fetus will be recorded. Fetuses will be individually identified with the study number, litter number and uterine distribution.

16.3. Visceral Examination Approximately one-half of the fetuses in each litter will be examined for visceral abnormalities by using a modification of the microdissection technique of Staples.2 Each fetus will be fixed in Bouin's solution, and the heads will subsequently be examined by free-hand sectioning;3 head sections will be stored in alcohol. The decapitated carcasses will not be retained.

16.4. Skeletal Examination The remaining fetuses (approximately one-half of the fetuses in each litter) will be examined for skeletal abnormalities after staining with alizarin red S.4 Following examination, skeletal preparations will be retained in glycerin with thymol added as a preservative.



17. COMPUTERIZED SYSTEMS The following proposed critical computerized systems may be used in the study. The actual computerized systems used will be specified in the Final Report.

Data for parameters not required by protocol, which are automatically generated by analytical devices used will be retained on file but not reported. Statistical analysis results that are generated by the program but are not required by protocol and/or are not scientifically relevant will be retained on file but will not be included in the tabulations.

Critical Computerized Systems

System Name Description of Data Collected and/or Analyzed Dispense Test article/vehicle/control article/component receipt

Argus Automated Data Collection and Management System

Clinical observations, body weights, food consumption, organ weights, Caesarean-sectioning observations, statistical analyses

Vivarium Temperature and Relative Humidity Monitoring System Study room temperature and humidity

Microsoft® Excel Pre- and postimplantation loss (report summary table) Quattro Pro Body weight graph

The SAS System Statistical analyses



18. STATISTICAL ANALYSIS Averages and percentages will be calculated. Litter values will be used where appropriate. Additional procedures and/or analyses may be performed, if appropriate.

A. Bartlett's Testc A. Kruskal-Wallis Test(≤75% ties)

I. Parametric II. Nonparametricb

Type of Testa

Significant at p<0.001 Significant at p<0.05Not Significant Not Significant

Nonparametric Dunn's TestAnalysis of Variance

Significant at p<0.05 Not Significant

Dunnett's Test

B. Fisher's Exact Test(>75% ties)

III. Test for Proportion Data Variance Test for Homogeneity

of the Binomial Distribution

a. Statistically significant probabilities are reported as either p<0.05 or p<0.01.b. Proportion data are not included in this category.c. Test for homogeneity of variance.



Clinical observations and other proportional data will be analyzed using the Variance Test for Homogeneity of the Binomial Distribution5. Continuous data (e.g., maternal body weights, body weight changes, food consumption values and litter averages for percent male fetuses, percent resorbed conceptuses, fetal body weights and fetal anomaly data) will be analyzed using Bartlett’s Test of Homogeneity of Variances6 and the Analysis of Variance7, when appropriate [i.e., Bartlett’s Test is not significant (p>0.001)]. If the Analysis of Variance is significant (p≤0.05), Dunnett’s Test8 will be used to identify the statistical significance of the individual groups. If the Analysis of Variance is not appropriate [i.e., Bartlett’s Test is significant (p≤0.001)], the Kruskal-Wallis Test9 is used (≤75% ties). In cases where the Kruskal-Wallis Test is statistically significant (p≤0.05), Dunn’s Method of Multiple Comparisons10 will be used to identify the statistical significance of the individual groups. If there are greater than 75% ties, Fisher’s Exact Test11 will be used to analyze the data.

Count data will be evaluated using the procedures described above for the Kruskal-Wallis Test9.

19. AMENDMENTS AND DEVIATIONS Changes to the approved protocol shall be made in the form of an amendment, which will be signed and dated by the Study Director and the Sponsor.

All protocol and SOP deviations will be documented in the study records. Deviations from the protocol and/or SOP related to the phaseof the study conducted at a Test Site shall be documented, acknowledged by the PI, and reported to the Study Director for authorization/acknowledgement. The Study Director will notify the Sponsor of deviations that may result in a significant impact on the study as soon as possible.

20. RETENTION OF RECORDS, SAMPLES, AND SPECIMENS All study-specific raw data, documentation, protocol, samples, specimens, and interim (if applicable) and final reports from this study are the property of the Sponsor. These materials will be available at the Testing Facility during the study and will be transferred to the Testing Facility archive by no later than the date of final report issue. One year after issue of the audited draft report, the Sponsor will be contacted to determine the disposition of materials associated with the study.

Electronic data generated by the Testing Facility will be archived, and the software and hardware required to produce it in a readable form will be maintained and available.

All records, samples, specimens and reports generated from phases or segments performed by Testing Facility-designated subcontractors will be returned to the Testing Facility for archiving.

Records to be maintained will include, but will not be limited to, documentation and data for the following:

• Protocol, protocol amendments, and deviations

• Study schedule



• Study-related correspondence

• Test and control article receipt, identification, preparation, and analysis

• Test system receipt, health, and husbandry

• Mating history

• Selection, assignment and replacement of animals

• Animal disposition

• Veterinary care, if required

• Administration of test and control articles

• In-life measurements and observations

• Gross and/or microscopic observations and related data

• Ovarian/Uterine and Fetal Observations

• Photographs, if required

• Packing and/or Shipment Lists

21. REPORTING A comprehensive Draft Report will be prepared following completion of the study and will be finalized following consultation with the Sponsor. The report will include all information necessary to provide a complete and accurate description of the experimental methods and results and any circumstances that may have affected the quality or integrity of the study.

The Sponsor will receive an electronic version of the Draft and Final Report provided in Adobe Acrobat PDF format (hyperlinked and searchable at final) along with a Microsoft Word version of the text. The PDF document will be created from native electronic files to the extent possible, including text and tables generated by the Testing Facility. Report components not available in native electronic files and/or original signature pages will be scanned and converted to PDF image files for incorporation. An original copy of the report with the Testing Facility’s handwritten signatures will be retained.

A tabulated data summary following the appropriate format as outlined in the ICH Harmonized Tripartite Guideline, The Common Technical Document for the Registration of Pharmaceuticals for Human Use: Safety – M4S (R2), Nonclinical Overview and Nonclinical Summaries of Module 2, Organisation of Module, will be provided as an appendix to the Draft and Final Reports.

Reports should be finalized within 6 months of issue of the audited Draft Report. If the Sponsor has not provided comments to the report within 6 months of draft issue, the report will be finalized by the Testing Facility unless other arrangements are made by the Sponsor.



22. ANIMAL WELFARE This study will comply with all applicable sections of the Final Rules of the Animal Welfare Act regulations (Code of Federal Regulations, Title 9), the Public Health Service Policy on Humane Care and Use of Laboratory Animals from the Office of Laboratory Animal Welfare, and the Guide for the Care and Use of Laboratory Animals from the National Research Council.1,12 The protocol and any amendments or procedures involving the care or use of animals in this study will be reviewed and approved by the Testing Facility Institutional Animal Care and Use Committee before the initiation of such procedures.

The Testing Facility’s attending veterinarian is responsible for implementation of programs for the evaluation of the health status of study animals, the recommendation of treatment for health conditions, the evaluation of response to treatment, as well as the diagnosis of pain or distress. The veterinary staff will communicate to the Study Director signs of animal illness, injury, pain and distress, and recommendations on treatment of the animal(s) and/or alteration of study procedures. In nonemergency situations, decisions regarding the study animals, including treatments, alterations in study design, and/or justification of action(s) will be documented, and will be approved in advance by the Study Director, and in consultation with the Sponsor as appropriate. If euthanasia is deemed necessary, it will be in accordance with the American Veterinary Medical Association (AVMA) Guidelines on Euthanasia and with the procedures outlined in the protocol.13 The Sponsor will be fully informed of any such events.

If an animal is determined by the veterinary staff to be in overt pain/distress, or appears moribund and is beyond the point where recovery appears reasonable, the animal will be euthanized for humane reasons in accordance with the AVMA Guidelines on Euthanasia.

By approving this protocol, the Sponsor affirms that there are no acceptable non-animal alternatives for this study, that this study is required by a relevant government regulatory agency(ies) and that it does not unnecessarily duplicate any previous experiments.



23. REFERENCES 1. National Research Council. Guide for the Care and Use of Laboratory Animals.

Washington, D.C.: National Academy Press. 1996.

2. Staples RE. Detection of visceral alterations in mammalian fetuses. Teratology 1974;9(3):A37-A38.

3. Wilson JG. Methods for administering agents and detecting malformations in experimental animals. In: Wilson JG, Warkany J, editors. Teratology: principles and techniques. Chicago (IL): University of Chicago Press; 1965. p. 262-77.

4. Staples RE, Schnell VL. Refinements in rapid clearing technic in the KOH-alizarin red S method for fetal bone. Stain Technol 1964;39:61-3.

5. Snedecor GW, Cochran WG. Variance test for homogeneity of the binomial distribution. Statistical methods. 6th Ed. Iowa State University Press, Ames; 1967. p. 240-1.

6. Sokal RR, Rohlf FJ. Bartlett' s test of homogeneity of variances. Biometry: the principles and practice of statistics in biological research. San Francisco (CA): Freeman & Co; 1969. p. 370-1.

7. Snedecor GW, Cochran WG. Analysis of variance. Statistical methods. 6th Ed. Iowa State University Press, Ames; 1967. p. 258-98.

8. Dunnett CW. A multiple comparison procedure for comparing several treatments with a control. J Am Stat Assoc 1955;50:1096-121.

9. Sokal RR, Rohlf FJ. Kruskal-Wallis test. Biometry: the principles and practice of statistics in biological research. San Francisco (CA): Freeman & Co; 1969. p. 388-91.

10. Dunn OJ. Multiple comparisons using rank sums. Technometrics 1964;6(3):241-52.

11. Siegel S. The Fisher’s exact probability test. Nonparametric statistics for the behavioral sciences. New York (NY): McGraw-Hill Co; 1956. p. 96-105.

12. Office of Laboratory Animal Welfare. Public Health Services Policy on Humane Care and Use of Laboratory Animals. Bethesda, MD: National Institutes of Health. August 2002.

13. American Veterinary Medical Association. AVMA Guidelines on Euthanasia. June 2007.



DEVIATIONS All deviations that occurred during the study have been authorized/acknowledged by the Study Director, assessed for impact, and documented in the study records. All protocol deviations that could have impacted the quality or integrity of the study are listed below.

Unless otherwise indicated below, none of the deviations were considered to have impacted the overall integrity of the study or the interpretation of the study results and conclusions.

Test and Control Articles

Samples retained on the last day of administration (28 May 2012) were submitted for analysis on 29 May 2012, rather than on the date prepared. This deviation did not adversely affect the outcome or interpretation of the study because the one day delay was immaterial; results of the analysis were within specification

The sample of the control article retained before the first dose administration was 5.5 mL (the contents of one vial), rather than 2.25 mL, as required by protocol. This deviation did not adversely affect the outcome or interpretation of the study because retention of the entire vial was appropriate for analysis.

Backup samples of the test and control articles were not retained before the first dose administration. This deviation did not adversely affect the outcome or interpretation of the study because analysis of backup samples was not required.

Environmental Conditions

Relative humidity was above the 30% to 70% range at one timepoint recorded throughout the course of the study. The online vivarium environmental monitoring system records temperature and humidity values no less frequently than four times hourly. The out-of-range humidity was 71.6%. This deviation did not adversely affect the outcome or interpretation of the study because this elevation in relative humidity was minor and transient.

In-life Observations, Measurements, and Evaluations

On 04 May 2012, during the acclimation period, a viability check was conducted only once, rather than twice daily. This deviation did not adversely affect the outcome or interpretation of the study because all rats were alive at the next viability check; rats were not yet assigned to study.

Necropsy

Fetus 4021-13 in the 0 (Control Article) mg/kg/day dose group was assigned to alcohol fixative and skeletal examination, but was placed into Bouin’s fixative and a visceral examination of the head was performed. A visceral examination of the body of this fetus was not performed. Fetus 4021-14 in the 0 (Control Article) mg/kg/day dose group was assigned to Bouin’s fixative and visceral examination, but was placed into alcohol after fresh visceral examination and was subsequently also examined for skeletal alterations. Fetus 4092-9 in the 180 mg/kg/day dose group was assigned to alcohol fixative and skeletal examination, but was placed into Bouin’s fixative and a visceral examination of the head and the fixed body of



this fetus was performed. These deviations did not adversely affect the outcome or interpretation of the study because approximately one-half of the fetuses in the litter were examined for each of visceral and skeletal alterations, as required by protocol.



Appendix 2 Test and Control Article Characterization


Revised Draft Report Page 101 Testing Facility Study No. 20024504

Appendix 3 Dose Formulation Analysis Report


FINAL REPORT

Study Phase: Analytical Chemistry


An Embryo-fetal Development Study of Thymosin beta 4 (formulated as RGN-352) by Intravenous Injection in Rats




Preclinical Services, Pennsylvania (PCS-PA) 905 Sheehy Dr., Building A


Page 1 of 42


Final Analytical Report Testing Facility Study No. 20024504

TABLE OF CONTENTS

1. LIST OF TABLES ..................................................................................................................3

2. LIST OF APPENDICES .........................................................................................................3

3. COMPLIANCE STATEMENT ..............................................................................................4

4. QUALITY ASSURANCE STATEMENT ..............................................................................5

5. RESPONSIBLE PERSONNEL...............................................................................................6

6. INTRODUCTION ...................................................................................................................7

7. EXPERIMENTAL DESIGN ...................................................................................................7

8. MATERIALS AND METHODS ............................................................................................7 8.1. Test Article.....................................................................................................................7 8.2. Control Article ...............................................................................................................8 8.3. Sample Receipt and Storage ..........................................................................................8 8.4. Sample Analysis.............................................................................................................8 8.5. Test and Control Article Inventory and Disposition ......................................................8 8.6. Computerized Systems...................................................................................................8

9. STATISTICAL ANALYSIS ...................................................................................................9

10. RETENTION OF RECORDS .................................................................................................9

11. RESULTS ................................................................................................................................9

12. CONCLUSION .....................................................................................................................10

13. REPORT APPROVAL ..........................................................................................................11



1. LIST OF TABLES

Table 1 Study Samples – Thymosin beta 4 Concentration .................................................12

2. LIST OF APPENDICES

Appendix 1 Deviations .............................................................................................................13

Appendix 2 Certificates of Analysis .........................................................................................15

Appendix 3 Analytical Procedure .............................................................................................20

Appendix 4 Dose Formulation Analysis Reports .....................................................................36



3. COMPLIANCE STATEMENT This phase of this study was conducted in accordance with the U.S. Department of Health and Human Services, Food and Drug Administration, United States Code of Federal Regulations, Title 21, Part 58: Good Laboratory Practice for Nonclinical Laboratory Studies and as accepted by Regulatory Authorities throughout the European Community (OECD Principles of Good Laboratory Practice) and Japan (MHLW)

Exceptions to GLPs include the following study elements:

• Characterization of the test and control articles was performed by a Sponsor subcontractor at a facility that follows FDA current Good Manufacturing Practice (cGMP) regulations. This exception to the GLP regulations did not affect the overall integrity of the study or the interpretation of the study results and conclusions because the material was appropriately characterized.

This phase of this study was conducted in accordance with the procedures described herein. All deviations authorized/acknowledged by the Principal Investigator and Study Director are documented in the Study Records. The report represents an accurate and complete record of the results obtained.



5. RESPONSIBLE PERSONNEL Principal Investigator Jason Sarsoza, BSc

Charles River Laboratories, PCS-PA

Research Assistant IV Phinh xu Ngo Charles River Laboratories, PCS-PA

Research Assistant II Shichei Andega, MS Charles River Laboratories, PCS-PA

Senior Manager, Laboratory Sciences Julian Gulbinski, III, BS, MBA Charles River Laboratories, PCS-PA

Director of Operations Matthew J. Vaneman, BS

Executive Director, Laboratory Sciences North America

Alan Bartlett, CChem, FRSC



6. INTRODUCTION The purpose of this project was to determine the concentration of Thymosin beta 4 in the test and control articles from Study No. 20024504 titled “An Embryo-fetal Development Study of Thymosin beta 4 (formulated as RGN-352) by Intravenous Injection in Rats.” The study was sponsored by RegeneRx Biopharmaceuticals, Inc., Rockville, MD where David Crockford, Vice President, served as the Sponsor Representative. Valerie A. Sharper, MS, Charles River Laboratories Preclinical Services, Horsham, PA, was the Study Director. Jason Sarsoza, BSc, Charles River Laboratories Preclinical Services, Horsham, PA, was the Principal Investigator for this study phase.

This study phase was started on 18 May 2012 and completed on 22 Jun 2012.

7. EXPERIMENTAL DESIGN Analysis of formulations was carried out with regard to concentration.

During the study, one vial each of Test Article and Control Article for analysis was taken at the start of study and end of study. The samples were received ambient and in satisfactory condition. The samples were stored 2°C to 8°C prior to analysis. The results are presented in Table 1.

8. MATERIALS AND METHODS

8.1. Test Article Identity: RGN-352 (also known as Thymosin beta 4 (Tβ4) Injectable

Solution)

Batch/Lot No.: 330-05-001

Expiration Date: 01 December 2012

Purity/Potency/Assay: 99.54% (assumed 100%)



Manufacturer/Supplier: Sponsor

The Sponsor provided to the Testing Facility documentation of the identity, strength, purity, composition, and stability for the test article, RGN-352. A Certificate of Analysis was provided to the Testing Facility and is presented in Appendix 2.



8.2. Control Article Identity: RGN-352 Placebo (also known as RGN-352 Vehicle)

Batch/Lot No.: 330-06-001

Expiration Date: 27 August 2012


Manufacturer/Supplier: Sponsor

The Sponsor provided to the Testing Facility documentation of the identity, strength, purity, composition, and stability for the control article, RGN-352 Placebo. A Certificate of Analysis was provided to the Testing Facility and is presented in Appendix 2.

8.3. Sample Receipt and Storage Two sets of samples, RGN-352 and RGN-352 Placebo, were received from the Testing Facility on 15 May 2012 and 29 May 2012. The samples were stored at 2°C ± 8°C prior to analysis. The samples were analyzed within the established stability period.

8.4. Sample Analysis Samples of RGN-352 and RGN-352 Placebo were analyzed for Thymosin beta 4 according to the validated method described in PCS-PA Analytical Procedure TYMN03 “Analytical Procedure for the Analysis of Thymosin Beta 4 in RGN-352 Placebo Dose Formulations by HPLC-UV.” A copy of the most recent version of the Analytical Procedure is contained in Appendix 3.

8.5. Test and Control Article Inventory and Disposition Records of the receipt, distribution, and storage of the test article (RGN-352) and control article (RGN-352 Placebo) were maintained. All unopened vials of test and control article were maintained at the Testing Facility.

8.6. Computerized Systems Critical computerized systems used in this study phase are listed below (see Text Table 1).

Text Table 1 Computerized Systems

System Name Version No. Description of Data Collected and/or Analyzed

TotalChrom® (PerkinElmer®) 6.2.1. Acquisition of HPLC data, assessment of system suitability, and integration of the peak area of the analyte

Excel (Microsoft Office) 1997 or later Regression analysis and calculation of concentrations and descriptive statistics



9. STATISTICAL ANALYSIS Regression analysis and descriptive statistics (such as means and relative standard deviations) were used to determine the dose formulation concentrations.

10. RETENTION OF RECORDS All study-specific raw data, documentation and the final report from this study phase are the property of the Sponsor. These materials will be available at the Testing Facility during the progress of the study and will be transferred to the Testing Facility archive by no later than the date of final report issue. One year after issue of the audited draft report, the Sponsor will be contacted to determine the disposition of materials associated with the study. Archival material will be indexed by Study No. 20024504.

11. RESULTS Concentration results are summarized in Table 1. Results and conclusions for each analytical run are provided in the Dose Formulation Analysis Reports, which are contained in Appendix 4.

All study samples analyzed had mean concentrations within or equal to the acceptance criteria of ± 10% (individual values within or equal to ± 15%) of their nominal concentrations.



12. CONCLUSION Samples of RGN-352 and RGN-352 Placebo were analyzed for Thymosin beta 4 by high-performance liquid chromatography with ultraviolet detection (HPLC-UV). The method was validated for the analysis of Thymosin beta 4 in RGN-352 Placebo at concentrations ranging from 50 mg/mL to 100 mg/mL.

The dose formulations were within specification.



Table 1 Study Samples – Thymosin beta 4 Concentration

Occasion (Sampling Date) Group

Theoretical Concentration

(mg/mL) Sampling Location

Measured Concentration

(mg/mL) Bias

Start of Study (15 May 2012)

1 0 Middle ND NA ND NA

Mean - NA

2 to 4 100 Middle 99.3 -0.7 99.9 -0.1

Mean 99.6 -0.4

End of Study (28 May 2012)

1 0 Middle ND NA ND NA

Mean - NA

2 to 4 100 Middle 101 1.0 99.2 -0.8

Mean 100 0.0 Group 1: Control article, RGN-352 Placebo Group 2-4: Test Article, RGN-352 NA = Not applicable. ND = None detected.



Appendix 1 Deviations



DEVIATIONS

All deviations that occurred during the study have been authorized/acknowledged by the Study Director, assessed for impact, and documented in the study records. Only minor SOP deviations that did not impact the quality or integrity of the study occurred during the course of the study.

None of the deviations were considered to have impacted the overall integrity of the study or the interpretation of the study results and conclusions.



Appendix 2 Certificates of Analysis



Appendix 3 Analytical Procedure



Appendix 4 Dose Formulation Analysis Reports



DOSE FORMULATION ANALYSIS REPORT

Sponsor: RegeneRx Biopharmacetuicals, Inc. Study Facility: Charles River Laboratories Preclinical Services, Pennsylvania Protocol Number: 20024504 Analyte: Thymosin beta 4 (formulated as RGN-352) Analytical Facility: Charles River Laboratories Preclinical Services, Pennsylvania Batch ID: 20024504-1-001-1 Sampling Criteria: Start of Study Concentration Analysis Vehicle: RGN-352 Placebo Storage Conditions: 2°C to 8°C Analytical Procedure: TYMN03 Revision 00 Analysis Date: May 18, 2012 RESULTS: (Concentrations in mg/mL, ND = none detected)

CALIBRATION STANDARDS

Standard Nominal Response Calculated % "X" = Criteria Standard Description Conc. Area Conc. Bias Exclude Limit Pass/Fail Cal Std A1 0.100 137207 0.0983 -1.7 5% PASS Cal Std B1 0.200 279280 0.199 -0.5 5% PASS Cal Std A2 0.400 565596 0.403 +0.8 5% PASS Cal Std B2 0.600 849985 0.605 +0.8 5% PASS Cal Std A3 0.800 1114811 0.793 -0.9 5% PASS Cal Std B3 1.00 1409064 1.00 0.0 5% PASS

CHECK STANDARDS

Standard Nominal Response Dilution Conc. % Criteria Standard Description Conc. Area Factor Found Bias Limit Pass/Fail

Check Std A3 0.800 1121892 1 0.798 -0.2 5% PASS Check Std A3 0.800 1113733 1 0.792 -1.0 5% PASS



Project Number: 20024504Analysis of Thymosin beta 4 in RGN-352

Batch ID: 20024504-1-001-1

0

200000

400000

600000

800000

1000000

1200000

1400000

1600000

0 0.2 0.4 0.6 0.8 1 1.2


Res

pons

e (A

rea)

Slope: 1407400Y-Int: -1149.2Corr: 0.99993

n: 6



SAMPLES

Nominal Total

Sample Prep Sample Response Dilution mg/mL % Description Date Conc. Replicate Area Factor Found Bias

Group 1 08/22/11 0 A 0 50.0 ND Group 1 08/22/11 0 B 0 50.0 ND

Groups 2 - 4 08/22/11 100 A 1117216 125 99.3 -0.7 Groups 2 - 4 08/22/11 100 B 1123088 125 99.9 -0.1

Sample

Description

Nominal Sample Conc.

Mean Conc.

%

Error Groups 2 - 4 100 99.6 -0.4

CONCLUSIONS: Results indicate that the formulations are within the acceptable limits of ±10% of nominal concentration for the mean result and ±15% of the nominal concentration for the individual results. ACTIONS TAKEN: None.



DOSE FORMULATION ANALYSIS REPORT

Sponsor: RegeneRx Biopharmaceuticals, Inc. Study Facility: Charles River Laboratories Preclinical Services, Pennsylvania Protocol Number: 20024504 Analyte: Thymosin beta 4 (formulated as RGN-352) Analytical Facility: Charles River Laboratories Preclinical Services, Pennsylvania Batch ID: 20024504-1-002-1 Sampling Criteria: End of Study Concentration Analysis Vehicle: RGN-352 Placebo Storage Conditions: 2°C to 8°C Analytical Procedure TYMN03 Revision 01 Analysis Date: June 21, 2012 RESULTS: (Concentrations in mg/mL, ND = none detected)

CALIBRATION STANDARDS

Standard Nominal Response Calculated % "X" = Criteria Standard Description Conc. Area Conc. Bias Exclude Limit Pass/Fail Cal Std A1 0.100 159307 0.102 +2.0 5% PASS Cal Std B1 0.200 305994 0.200 0.0 5% PASS Cal Std A2 0.400 612207 0.404 +1.0 5% PASS Cal Std B2 0.600 892435 0.590 -1.7 5% PASS Cal Std A3 0.800 1208114 0.800 0.0 5% PASS Cal Std B3 1.00 1514957 1.00 0.0 5% PASS

CHECK STANDARDS

Standard Nominal Response Dilution Conc. % Criteria Standard Description Conc. Area Factor Found Bias Limit Pass/Fail

Check Std A3 0.800 1215935 1 0.805 +0.6 5% PASS Check Std A3 0.800 1219181 1 0.807 +0.9 5% PASS



Project Number: 20024504Analysis of Thymosin beta 4 in RGN-352

Batch ID: 20024504-1-002-1

0

200000

400000

600000

800000

1000000

1200000

1400000

1600000

0 0.2 0.4 0.6 0.8 1 1.2


Res

pons

e (A

rea)

Slope: 1502800Y-Int: 5727.2

Corr: 0.99989n: 6



SAMPLES

Nominal Total Sample Prep Sample Response Dilution mg/mL %

Description Date Conc. Replicate Area Factor Found Bias Group 1 08/22/11 0 A 0 50.0 ND Group 1 08/22/11 0 B 0 50.0 ND

Group 2-4 08/22/11 100 A 1223049 125 101 +1.0 Group 2-4 08/22/11 100 B 1198381 125 99.2 -0.8

Sample

Description

Nominal Sample Conc.

Mean Conc.

%

Error Groups 2-4 100 100 0.0

CONCLUSIONS: Results indicate that the formulations are within the acceptable limits of ±10% of nominal concentrations for the mean result and ±15% of the nominal concentration for the individual results. ACTIONS TAKEN: None.



Appendix 4 Clinical Observations - Individual Data


Revised Draft Report Page 144 Testing Facility Study No. 20024504 PROTOCOL 20024504: AN EMBRYO-FETAL DEVELOPMENT STUDY OF THYMOSIN BETA 4 (FORMULATED AS RGN-532) BY INTRAVENOUS INJECTION IN RATS APPENDIX 4 (PAGE 1): CLINICAL OBSERVATIONS - INDIVIDUAL DATA ------------------------------------------------------------------------------------------------------------------------------------ GROUP 1 CONTROL ARTICLE 0 (CONTROL) MG/KG/DAY ------------------------------------------------------------------------------------------------------------------------------------ RAT # DESCRIPTION ------------------------------------------------------------------------------------------------------------------------------------ 4001 NO ADVERSE FINDINGS 4002 NO ADVERSE FINDINGS 4003 NO ADVERSE FINDINGS 4004 NO ADVERSE FINDINGS 4005 NO ADVERSE FINDINGS 4006 NO ADVERSE FINDINGS 4007 NO ADVERSE FINDINGS 4008 NO ADVERSE FINDINGS 4009 NO ADVERSE FINDINGS 4010 NO ADVERSE FINDINGS 4011 NO ADVERSE FINDINGS 4012 NO ADVERSE FINDINGS 4013 NO ADVERSE FINDINGS 4014 NO ADVERSE FINDINGS 4015 NO ADVERSE FINDINGS 4016 NO ADVERSE FINDINGS 4017 DG( 21 ) SPARSE HAIR COAT: LIMB(S)a 4018 NO ADVERSE FINDINGS 4019 NO ADVERSE FINDINGS 4020 NO ADVERSE FINDINGS 4021 NO ADVERSE FINDINGS 4022 NO ADVERSE FINDINGS 4023 NO ADVERSE FINDINGS 4024 NO ADVERSE FINDINGS 4025 NO ADVERSE FINDINGS ------------------------------------------------------------------------------------------------------------------------------------ CLINICAL OBSERVATIONS APPEARED NORMAL UNLESS NOTED OTHERWISE DG = DAY OF PRESUMED GESTATION a. Observation confirmed at necropsy.


Revised Draft Report Page 145 Testing Facility Study No. 20024504 PROTOCOL 20024504: AN EMBRYO-FETAL DEVELOPMENT STUDY OF THYMOSIN BETA 4 (FORMULATED AS RGN-532) BY INTRAVENOUS INJECTION IN RATS APPENDIX 4 (PAGE 2): CLINICAL OBSERVATIONS - INDIVIDUAL DATA ------------------------------------------------------------------------------------------------------------------------------------ GROUP 2 RGN-352 20 MG/KG/DAY ------------------------------------------------------------------------------------------------------------------------------------ RAT # DESCRIPTION ------------------------------------------------------------------------------------------------------------------------------------ 4026 NO ADVERSE FINDINGS 4027 NO ADVERSE FINDINGS 4028 NO ADVERSE FINDINGS 4029 NO ADVERSE FINDINGS 4030 NO ADVERSE FINDINGS 4031 NO ADVERSE FINDINGS 4032 NO ADVERSE FINDINGS 4033 NO ADVERSE FINDINGS 4034 NO ADVERSE FINDINGS 4035 NO ADVERSE FINDINGS 4036 NO ADVERSE FINDINGS 4037 NO ADVERSE FINDINGS 4038 NO ADVERSE FINDINGS 4039 NO ADVERSE FINDINGS 4040 NO ADVERSE FINDINGS 4041 NO ADVERSE FINDINGS 4042 NO ADVERSE FINDINGS 4043 NO ADVERSE FINDINGS 4044 NO ADVERSE FINDINGS 4045 NO ADVERSE FINDINGS 4046 NO ADVERSE FINDINGS 4047 NO ADVERSE FINDINGS 4048 NO ADVERSE FINDINGS 4049 NO ADVERSE FINDINGS 4050 NO ADVERSE FINDINGS ------------------------------------------------------------------------------------------------------------------------------------ CLINICAL OBSERVATIONS APPEARED NORMAL UNLESS NOTED OTHERWISE DG = DAY OF PRESUMED GESTATION


Revised Draft Report Page 146 Testing Facility Study No. 20024504 PROTOCOL 20024504: AN EMBRYO-FETAL DEVELOPMENT STUDY OF THYMOSIN BETA 4 (FORMULATED AS RGN-532) BY INTRAVENOUS INJECTION IN RATS APPENDIX 4 (PAGE 3): CLINICAL OBSERVATIONS - INDIVIDUAL DATA ------------------------------------------------------------------------------------------------------------------------------------ GROUP 3 RGN-352 60 MG/KG/DAY ------------------------------------------------------------------------------------------------------------------------------------ RAT # DESCRIPTION ------------------------------------------------------------------------------------------------------------------------------------ 4051 NO ADVERSE FINDINGS 4052 NO ADVERSE FINDINGS 4053 NO ADVERSE FINDINGS 4054 NO ADVERSE FINDINGS 4055 NO ADVERSE FINDINGS 4056 NO ADVERSE FINDINGS 4057 NO ADVERSE FINDINGS 4058 NO ADVERSE FINDINGS 4059 NO ADVERSE FINDINGS 4060 NO ADVERSE FINDINGS 4061 NO ADVERSE FINDINGS 4062 NO ADVERSE FINDINGS 4063 NO ADVERSE FINDINGS 4064 NO ADVERSE FINDINGS 4065 NO ADVERSE FINDINGS 4066 NO ADVERSE FINDINGS 4067 NO ADVERSE FINDINGS 4068 NO ADVERSE FINDINGS 4069 NO ADVERSE FINDINGS 4070 NO ADVERSE FINDINGS 4071 NO ADVERSE FINDINGS 4072 NO ADVERSE FINDINGS 4073 NO ADVERSE FINDINGS 4074 NO ADVERSE FINDINGS 4075 NO ADVERSE FINDINGS ------------------------------------------------------------------------------------------------------------------------------------ CLINICAL OBSERVATIONS APPEARED NORMAL UNLESS NOTED OTHERWISE DG = DAY OF PRESUMED GESTATION


Revised Draft Report Page 147 Testing Facility Study No. 20024504 PROTOCOL 20024504: AN EMBRYO-FETAL DEVELOPMENT STUDY OF THYMOSIN BETA 4 (FORMULATED AS RGN-532) BY INTRAVENOUS INJECTION IN RATS APPENDIX 4 (PAGE 4): CLINICAL OBSERVATIONS - INDIVIDUAL DATA ------------------------------------------------------------------------------------------------------------------------------------ GROUP 4 RGN-352 180 MG/KG/DAY ------------------------------------------------------------------------------------------------------------------------------------ RAT # DESCRIPTION ------------------------------------------------------------------------------------------------------------------------------------ 4076 NO ADVERSE FINDINGS 4077 NO ADVERSE FINDINGS 4078 NO ADVERSE FINDINGS 4079 NO ADVERSE FINDINGS 4080 DG( 11- 21) PORTION OF TAIL MISSING a 4081 NO ADVERSE FINDINGS 4082 NO ADVERSE FINDINGS 4083 NO ADVERSE FINDINGS 4084 NO ADVERSE FINDINGS 4085 NO ADVERSE FINDINGS 4086 NO ADVERSE FINDINGS 4087 NO ADVERSE FINDINGS 4088 NO ADVERSE FINDINGS 4089 DG( 21 ) SPARSE HAIR COAT: NECK a 4090 NO ADVERSE FINDINGS 4091 NO ADVERSE FINDINGS 4092 NO ADVERSE FINDINGS 4093 NO ADVERSE FINDINGS 4094 NO ADVERSE FINDINGS 4095 NO ADVERSE FINDINGS 4096 NO ADVERSE FINDINGS 4097 DG( 19- 21) SPARSE HAIR COAT: UNDERSIDE a DG( 21 ) SPARSE HAIR COAT: LIMB(S)a 4098 NO ADVERSE FINDINGS 4099 NO ADVERSE FINDINGS 4100 NO ADVERSE FINDINGS ------------------------------------------------------------------------------------------------------------------------------------ CLINICAL OBSERVATIONS APPEARED NORMAL UNLESS NOTED OTHERWISE DG = DAY OF PRESUMED GESTATION a. Observation confirmed at necropsy.



Appendix 5 Maternal Body Weights - Individual Data


Revised Draft Report Page 149 Testing Facility Study No. 20024504 PROTOCOL 20024504: AN EMBRYO-FETAL DEVELOPMENT STUDY OF THYMOSIN BETA 4 (FORMULATED AS RGN-532) BY INTRAVENOUS INJECTION IN RATS APPENDIX 5 (PAGE 1): MATERNAL BODY WEIGHTS - INDIVIDUAL DATA ------------------------------------------------------------------------------------------------------------------------------------ PREGNANCY STATUS DAY 0 7 8 9 10 11 12 13 14 15 16 17 18 ------------------------------------------------------------------------------------------------------------------------------------ RAT # GROUP 1 CONTROL ARTICLE 0 (CONTROL) MG/KG/DAY ------------------------------------------------------------------------------------------------------------------------------------ 4001 P 241. 283. 296. 295. 304. 308. 312. 315. 324. 328. 342. 359. 375. 4002 P 230. 280. 289. 290. 297. 302. 307. 310. 317. 320. 331. 348. 356. 4003 P 242. 288. 290. 296. 305. 310. 315. 321. 327. 335. 344. 368. 378. 4004 P 244. 306. 308. 312. 318. 329. 330. 337. 346. 352. 365. 377. 392. 4005 P 252. 294. 293. 292. 298. 309. 308. 315. 316. 322. 335. 357. 372. 4006 P 252. 310. 316. 318. 329. 341. 341. 346. 359. 364. 372. 400. 404. 4007 P 247. 284. 295. 298. 310. 318. 317. 328. 331. 340. 357. 362. 383. 4008 P 228. 266. 268. 272. 274. 282. 286. 291. 294. 300. 315. 331. 344. 4009 P 248. 299. 303. 305. 311. 324. 325. 330. 338. 352. 368. 379. 396. 4010 P 241. 275. 280. 287. 293. 301. 303. 304. 306. 310. 326. 342. 345. 4011 P 250. 290. 283. 288. 292. 302. 303. 308. 320. 322. 327. 342. 368. 4012 P 248. 292. 286. 296. 307. 309. 312. 321. 328. 332. 338. 350. 368. 4013 P 246. 294. 288. 303. 308. 307. 318. 315. 322. 326. 339. 345. 364. 4014 P 240. 265. 271. 279. 282. 290. 292. 301. 304. 311. 331. 330. 349. 4015 P 232. 268. 260. 266. 270. 274. 284. 285. 281. 288. 292. 304. 328. 4016 P 249. 292. 284. 290. 292. 300. 309. 309. 314. 328. 344. 353. 369. 4017 P 258. 310. 314. 316. 329. 334. 342. 346. 357. 363. 378. 395. 412. 4018 P 244. 288. 297. 299. 305. 307. 319. 313. 326. 328. 342. 361. 376. 4019 P 242. 271. 277. 275. 281. 286. 294. 299. 302. 312. 315. 330. 337. 4020 P 237. 275. 275. 282. 284. 285. 298. 303. 311. 317. 326. 341. 355. 4021 P 235. 274. 281. 288. 292. 300. 301. 312. 314. 322. 336. 355. 365. 4022 P 238. 282. 290. 298. 302. 307. 312. 318. 322. 333. 340. 354. 366. 4023 P 234. 286. 282. 294. 295. 300. 305. 318. 320. 331. 340. 352. 367. 4024 P 243. 287. 285. 286. 289. 302. 306. 311. 312. 319. 329. 342. 353. 4025 P 234. 275. 274. 277. 278. 286. 285. 289. 297. 298. 306. 312. 323. ------------------------------------------------------------------------------------------------------------------------------------ P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G).


Revised Draft Report Page 150 Testing Facility Study No. 20024504 PROTOCOL 20024504: AN EMBRYO-FETAL DEVELOPMENT STUDY OF THYMOSIN BETA 4 (FORMULATED AS RGN-532) BY INTRAVENOUS INJECTION IN RATS APPENDIX 5 (PAGE 2): MATERNAL BODY WEIGHTS - INDIVIDUAL DATA ------------------------------------------------------------------------------------------------------------------------------------ PREGNANCY STATUS DAY 19 20 21 ------------------------------------------------------------------------------------------------------------------------------------ RAT # GROUP 1 CONTROL ARTICLE 0 (CONTROL) MG/KG/DAY ------------------------------------------------------------------------------------------------------------------------------------ 4001 P 383. 407. 435. 4002 P 372. 383. 413. 4003 P 392. 414. 440. 4004 P 398. 425. 474. 4005 P 388. 407. 440. 4006 P 427. 443. 478. 4007 P 401. 421. 450. 4008 P 358. 373. 409. 4009 P 413. 430. 455. 4010 P 357. 376. 399. 4011 P 383. 398. 428. 4012 P 385. 405. 433. 4013 P 378. 402. 428. 4014 P 361. 382. 414. 4015 P 335. 352. 368. 4016 P 389. 409. 436. 4017 P 429. 450. 485. 4018 P 396. 416. 450. 4019 P 352. 366. 389. 4020 P 371. 388. 410. 4021 P 379. 402. 450. 4022 P 383. 400. 427. 4023 P 375. 396. 420. 4024 P 365. 376. 402. 4025 P 336. 350. 376. ------------------------------------------------------------------------------------------------------------------------------------ P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G).


Revised Draft Report Page 151 Testing Facility Study No. 20024504 PROTOCOL 20024504: AN EMBRYO-FETAL DEVELOPMENT STUDY OF THYMOSIN BETA 4 (FORMULATED AS RGN-532) BY INTRAVENOUS INJECTION IN RATS APPENDIX 5 (PAGE 3): MATERNAL BODY WEIGHTS - INDIVIDUAL DATA ------------------------------------------------------------------------------------------------------------------------------------ PREGNANCY STATUS DAY 0 7 8 9 10 11 12 13 14 15 16 17 18 ------------------------------------------------------------------------------------------------------------------------------------ RAT # GROUP 2 RGN-352 20 MG/KG/DAY ------------------------------------------------------------------------------------------------------------------------------------ 4026 P 238. 276. 282. 284. 289. 295. 296. 304. 310. 313. 318. 338. 346. 4027 P 251. 291. 294. 291. 298. 311. 309. 311. 315. 323. 328. 349. 370. 4028 P 239. 281. 277. 290. 290. 296. 297. 301. 304. 310. 312. 324. 331. 4029 P 250. 290. 295. 298. 300. 308. 311. 318. 321. 327. 338. 358. 370. 4030 P 246. 283. 290. 290. 298. 305. 310. 313. 317. 321. 332. 341. 356. 4031 P 247. 288. 286. 294. 307. 314. 312. 320. 326. 335. 345. 361. 377. 4032 P 235. 272. 274. 274. 275. 278. 281. 289. 294. 296. 308. 324. 341. 4033 P 242. 300. 306. 310. 317. 326. 321. 331. 338. 346. 362. 376. 397. 4034 P 248. 309. 315. 313. 321. 328. 335. 336. 342. 359. 366. 382. 374. 4035 P 241. 289. 291. 295. 296. 304. 308. 309. 318. 324. 341. 351. 369. 4036 P 252. 309. 315. 315. 328. 327. 311. 344. 348. 360. 373. 389. 410. 4037 P 233. 263. 267. 272. 273. 280. 280. 284. 289. 294. 302. 318. 328. 4038 P 240. 294. 292. 298. 298. 305. 303. 320. 321. 322. 340. 348. 364. 4039 P 242. 282. 288. 294. 298. 302. 312. 314. 321. 328. 340. 350. 368. 4040 P 243. 289. 288. 295. 302. 301. 312. 313. 300. 315. 326. 331. 353. 4041 P 252. 293. 296. 299. 307. 337. 318. 321. 328. 338. 354. 366. 383. 4042 P 243. 278. 282. 290. 295. 304. 306. 308. 318. 328. 347. 356. 369. 4043 P 244. 280. 274. 280. 292. 287. 290. 298. 301. 314. 320. 329. 351. 4044 P 236. 278. 282. 291. 296. 302. 311. 316. 322. 330. 342. 362. 382. 4045 P 230. 268. 268. 275. 279. 281. 289. 291. 293. 304. 308. 314. 331. 4046 P 235. 278. 279. 284. 287. 290. 288. 291. 297. 309. 312. 323. 328. 4047 P 258. 305. 312. 316. 322. 331. 336. 339. 345. 357. 368. 386. 406. 4048 P 227. 270. 272. 280. 284. 293. 296. 302. 308. 321. 329. 348. 365. 4049 P 241. 280. 278. 280. 287. 293. 296. 300. 312. 315. 326. 340. 363. 4050 P 232. 270. 271. 272. 275. 280. 280. 284. 294. 300. 309. 323. 336. ------------------------------------------------------------------------------------------------------------------------------------ P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G).


Revised Draft Report Page 152 Testing Facility Study No. 20024504 PROTOCOL 20024504: AN EMBRYO-FETAL DEVELOPMENT STUDY OF THYMOSIN BETA 4 (FORMULATED AS RGN-532) BY INTRAVENOUS INJECTION IN RATS APPENDIX 5 (PAGE 4): MATERNAL BODY WEIGHTS - INDIVIDUAL DATA ------------------------------------------------------------------------------------------------------------------------------------ PREGNANCY STATUS DAY 19 20 21 ------------------------------------------------------------------------------------------------------------------------------------ RAT # GROUP 2 RGN-352 20 MG/KG/DAY ------------------------------------------------------------------------------------------------------------------------------------ 4026 P 371. 381. 403. 4027 P 386. 398. 427. 4028 P 345. 357. 373. 4029 P 386. 396. 434. 4030 P 375. 382. 405. 4031 P 386. 416. 437. 4032 P 357. 380. 413. 4033 P 412. 442. 481. 4034 P 392. 416. 445. 4035 P 381. 397. 426. 4036 P 434. 450. 491. 4037 P 346. 356. 388. 4038 P 382. 399. 426. 4039 P 383. 397. 429. 4040 P 366. 380. 409. 4041 P 401. 418. 456. 4042 P 383. 406. 439. 4043 P 360. 374. 382. 4044 P 402. 422. 462. 4045 P 331. 346. 374. 4046 P 340. 356. 386. 4047 P 424. 443. 464. 4048 P 382. 409. 436. 4049 P 382. 402. 432. 4050 P 360. 371. 396. ------------------------------------------------------------------------------------------------------------------------------------ P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G).


Revised Draft Report Page 153 Testing Facility Study No. 20024504 PROTOCOL 20024504: AN EMBRYO-FETAL DEVELOPMENT STUDY OF THYMOSIN BETA 4 (FORMULATED AS RGN-532) BY INTRAVENOUS INJECTION IN RATS APPENDIX 5 (PAGE 5): MATERNAL BODY WEIGHTS - INDIVIDUAL DATA ------------------------------------------------------------------------------------------------------------------------------------ PREGNANCY STATUS DAY 0 7 8 9 10 11 12 13 14 15 16 17 18 ------------------------------------------------------------------------------------------------------------------------------------ RAT # GROUP 3 RGN-352 60 MG/KG/DAY ------------------------------------------------------------------------------------------------------------------------------------ 4051 P 232. 271. 272. 274. 277. 285. 291. 298. 304. 311. 315. 327. 344. 4052 P 230. 269. 268. 274. 279. 288. 294. 303. 310. 312. 328. 341. 355. 4053 P 243. 283. 283. 280. 291. 295. 301. 302. 302. 318. 325. 342. 357. 4054 P 251. 306. 310. 312. 321. 329. 334. 339. 346. 349. 359. 366. 379. 4055 P 242. 296. 293. 295. 306. 314. 317. 328. 334. 340. 350. 368. 380. 4056 P 239. 283. 284. 288. 295. 302. 309. 310. 318. 322. 331. 338. 357. 4057 P 229. 268. 271. 267. 276. 277. 281. 287. 293. 299. 304. 312. 328. 4058 P 235. 274. 279. 280. 286. 287. 294. 299. 308. 314. 323. 335. 342. 4059 P 242. 301. 304. 306. 314. 322. 320. 325. 332. 338. 343. 362. 366. 4060 P 240. 281. 280. 283. 289. 300. 301. 306. 313. 323. 336. 349. 370. 4061 P 244. 289. 288. 296. 306. 314. 317. 328. 330. 336. 350. 360. 375. 4062 P 248. 300. 295. 302. 307. 322. 322. 325. 332. 337. 343. 359. 384. 4063 P 236. 294. 298. 304. 310. 319. 330. 332. 338. 345. 360. 377. 398. 4064 P 249. 279. 278. 283. 288. 296. 299. 303. 308. 317. 327. 342. 356. 4065 P 245. 283. 284. 289. 297. 302. 306. 313. 318. 327. 340. 348. 365. 4066 P 258. 303. 304. 305. 311. 326. 332. 339. 349. 358. 379. 390. 405. 4067 P 241. 284. 289. 296. 299. 304. 311. 317. 323. 336. 342. 351. 363. 4068 P 246. 276. 282. 286. 294. 294. 303. 302. 309. 321. 329. 344. 353. 4069 P 253. 289. 291. 291. 302. 313. 315. 322. 328. 341. 351. 362. 379. 4070 P 238. 279. 295. 297. 301. 308. 314. 322. 324. 334. 346. 344. 364. 4071 P 241. 289. 293. 304. 298. 307. 310. 313. 314. 331. 345. 352. 370. 4072 P 235. 286. 292. 301. 302. 310. 314. 322. 325. 338. 348. 364. 373. 4073 P 246. 288. 294. 292. 300. 308. 309. 322. 322. 331. 342. 355. 368. 4074 P 252. 295. 296. 301. 306. 301. 309. 315. 312. 308. 328. 333. 350. 4075 P 232. 284. 281. 287. 296. 300. 304. 308. 318. 319. 333. 339. 362. ------------------------------------------------------------------------------------------------------------------------------------ P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G).


Revised Draft Report Page 154 Testing Facility Study No. 20024504 PROTOCOL 20024504: AN EMBRYO-FETAL DEVELOPMENT STUDY OF THYMOSIN BETA 4 (FORMULATED AS RGN-532) BY INTRAVENOUS INJECTION IN RATS APPENDIX 5 (PAGE 6): MATERNAL BODY WEIGHTS - INDIVIDUAL DATA ------------------------------------------------------------------------------------------------------------------------------------ PREGNANCY STATUS DAY 19 20 21 ------------------------------------------------------------------------------------------------------------------------------------ RAT # GROUP 3 RGN-352 60 MG/KG/DAY ------------------------------------------------------------------------------------------------------------------------------------ 4051 P 365. 378. 408. 4052 P 373. 388. 420. 4053 P 366. 380. 406. 4054 P 402. 409. 446. 4055 P 400. 413. 435. 4056 P 368. 388. 417. 4057 P 353. 370. 397. 4058 P 350. 366. 395. 4059 P 385. 391. 424. 4060 P 392. 411. 435. 4061 P 389. 410. 445. 4062 P 399. 417. 444. 4063 P 418. 438. 475. 4064 P 369. 390. 422. 4065 P 381. 405. 425. 4066 P 424. 440. 474. 4067 P 382. 403. 436. 4068 P 376. 393. 422. 4069 P 400. 422. 444. 4070 P 379. 394. 425. 4071 P 385. 409. 442. 4072 P 388. 410. 429. 4073 P 389. 405. 438. 4074 P 358. 375. 395. 4075 P 383. 400. 428. ------------------------------------------------------------------------------------------------------------------------------------ P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G).


Revised Draft Report Page 155 Testing Facility Study No. 20024504 PROTOCOL 20024504: AN EMBRYO-FETAL DEVELOPMENT STUDY OF THYMOSIN BETA 4 (FORMULATED AS RGN-532) BY INTRAVENOUS INJECTION IN RATS APPENDIX 5 (PAGE 7): MATERNAL BODY WEIGHTS - INDIVIDUAL DATA ------------------------------------------------------------------------------------------------------------------------------------ PREGNANCY STATUS DAY 0 7 8 9 10 11 12 13 14 15 16 17 18 ------------------------------------------------------------------------------------------------------------------------------------ RAT # GROUP 4 RGN-352 180 MG/KG/DAY ------------------------------------------------------------------------------------------------------------------------------------ 4076 P 240. 293. 294. 299. 310. 314. 320. 329. 334. 342. 355. 366. 378. 4077 NP 231. 248. 243. 241. 245. 252. 245. 247. 251. 254. 253. 256. 255. 4078 P 250. 294. 299. 303. 313. 317. 318. 326. 330. 336. 345. 359. 366. 4079 P 236. 280. 288. 290. 295. 299. 305. 309. 319. 331. 341. 362. 375. 4080 P 248. 288. 301. 298. 304. 315. 315. 322. 325. 329. 342. 362. 374. 4081 P 231. 267. 271. 274. 278. 283. 290. 290. 297. 308. 316. 329. 345. 4082 P 246. 286. 291. 296. 298. 303. 313. 313. 319. 332. 342. 359. 375. 4083 P 242. 280. 281. 286. 292. 300. 300. 308. 310. 318. 329. 337. 356. 4084 P 228. 267. 265. 270. 277. 283. 289. 293. 298. 306. 324. 336. 350. 4085 P 244. 290. 293. 302. 304. 313. 313. 321. 326. 330. 340. 354. 374. 4086 P 245. 286. 289. 294. 299. 300. 304. 311. 313. 322. 333. 343. 353. 4087 P 247. 288. 294. 299. 309. 314. 322. 332. 335. 344. 359. 374. 396. 4088 P 251. 296. 290. 298. 308. 315. 328. 333. 348. 355. 372. 387. 402. 4089 P 240. 281. 284. 287. 299. 304. 305. 316. 316. 319. 341. 350. 363. 4090 P 246. 293. 298. 298. 308. 316. 326. 330. 337. 342. 350. 362. 378. 4091 P 243. 294. 292. 297. 306. 314. 314. 316. 323. 336. 348. 365. 389. 4092 P 253. 303. 308. 310. 320. 321. 323. 329. 334. 347. 352. 362. 376. 4093 P 242. 285. 291. 292. 296. 302. 308. 316. 320. 323. 335. 346. 366. 4094 P 241. 293. 294. 301. 310. 315. 315. 325. 327. 346. 349. 369. 386. 4095 P 252. 310. 316. 324. 328. 333. 339. 344. 349. 351. 361. 379. 395. 4096 P 234. 276. 278. 277. 287. 288. 297. 300. 305. 310. 324. 341. 358. 4097 P 238. 279. 275. 275. 278. 286. 295. 297. 303. 307. 318. 334. 352. 4098 NP 242. 280. 275. 286. 288. 288. 291. 294. 284. 280. 280. 283. 278. 4099 P 258. 315. 308. 317. 318. 323. 333. 337. 340. 349. 371. 389. 407. 4100 P 233. 278. 280. 286. 291. 291. 297. 298. 311. 319. 331. 346. 362. ------------------------------------------------------------------------------------------------------------------------------------ P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G).


Revised Draft Report Page 156 Testing Facility Study No. 20024504 PROTOCOL 20024504: AN EMBRYO-FETAL DEVELOPMENT STUDY OF THYMOSIN BETA 4 (FORMULATED AS RGN-532) BY INTRAVENOUS INJECTION IN RATS APPENDIX 5 (PAGE 8): MATERNAL BODY WEIGHTS - INDIVIDUAL DATA ------------------------------------------------------------------------------------------------------------------------------------ PREGNANCY STATUS DAY 19 20 21 ------------------------------------------------------------------------------------------------------------------------------------ RAT # GROUP 4 RGN-352 180 MG/KG/DAY ------------------------------------------------------------------------------------------------------------------------------------ 4076 P 396. 409. 443. 4077 NP 256. 257. 260. 4078 P 384. 400. 426. 4079 P 396. 417. 440. 4080 P 373. 392. 415. 4081 P 359. 380. 406. 4082 P 395. 418. 447. 4083 P 371. 392. 429. 4084 P 364. 378. 415. 4085 P 395. 418. 440. 4086 P 370. 383. 415. 4087 P 415. 433. 467. 4088 P 423. 442. 470. 4089 P 384. 396. 421. 4090 P 394. 404. 427. 4091 P 408. 426. 470. 4092 P 396. 412. 435. 4093 P 380. 394. 413. 4094 P 396. 419. 458. 4095 P 415. 433. 469. 4096 P 375. 392. 417. 4097 P 367. 386. 415. 4098 NP 274. 279. 288. 4099 P 421. 446. 482. 4100 P 379. 402. 430. ------------------------------------------------------------------------------------------------------------------------------------ P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G).



Appendix 6 Maternal Food Consumption Values - Individual Data


Revised Draft Report Page 158 Testing Facility Study No. 20024504 PROTOCOL 20024504: AN EMBRYO-FETAL DEVELOPMENT STUDY OF THYMOSIN BETA 4 (FORMULATED AS RGN-532) BY INTRAVENOUS INJECTION IN RATS APPENDIX 6 (PAGE 1): MATERNAL FOOD CONSUMPTION VALUES - INDIVIDUAL DATA ------------------------------------------------------------------------------------------------------------------------------------ PREGNANCY STATUS DAYS 0 - 7 7 - 10 10 - 12 12 - 15 15 - 18 18 - 21 ------------------------------------------------------------------------------------------------------------------------------------ RAT # GROUP 1 CONTROL ARTICLE 0 (CONTROL) MG/KG/DAY ------------------------------------------------------------------------------------------------------------------------------------ 4001 P 145. 65. 47. 70. 86. 86. 4002 P 149. 69. 47. 73. 82. 85. 4003 P 157. 70. 50. 76. 90. 90. 4004 P 193. 77. 54. 88. 72. 93. 4005 P 160. 66. 49. 74. 85. 86. 4006 P 189. 84. 60. 98. 103. 104. 4007 P 135. 69. 49. 80. 84. 95. 4008 P 131. 57. 40. 67. 77. 72. 4009 P 180. 73. 60. 86. 88. 92. 4010 P 141. 70. 47. 66. 74. 70. 4011 P 159. 69. 48. 73. 74. 71. 4012 P 164. 68. 50. 77. 79. 87. 4013 P 163. 68. 45. 73. 72. 79. 4014 P 144. 73. 54. 77. 86. 92. 4015 P 128. 56. 40. 51. 64. 72. 4016 P 155. 60. 47. 73. 80. 81. 4017 P 164. 76. 54. 86. 93. 98. 4018 P 155. 73. 49. 69. 82. 91. 4019 P 124. 56. 42. 63. 71. 73. 4020 P 146. 63. 49. 75. 91. 88. 4021 P 142. 67. 49. 66. 78. 89. 4022 P 142. 68. 47. 74. 74. 81. 4023 P 145. 72. 51. 78. 86. 91. 4024 P 149. 65. 46. 68. 74. 79. 4025 P 148. 67. 48. 74. 80. 78. ------------------------------------------------------------------------------------------------------------------------------------ P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAYS = DAYS OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G).


Revised Draft Report Page 159 Testing Facility Study No. 20024504 PROTOCOL 20024504: AN EMBRYO-FETAL DEVELOPMENT STUDY OF THYMOSIN BETA 4 (FORMULATED AS RGN-532) BY INTRAVENOUS INJECTION IN RATS APPENDIX 6 (PAGE 2): MATERNAL FOOD CONSUMPTION VALUES - INDIVIDUAL DATA ------------------------------------------------------------------------------------------------------------------------------------ PREGNANCY STATUS DAYS 0 - 7 7 - 10 10 - 12 12 - 15 15 - 18 18 - 21 ------------------------------------------------------------------------------------------------------------------------------------ RAT # GROUP 2 RGN-352 20 MG/KG/DAY ------------------------------------------------------------------------------------------------------------------------------------ 4026 P 147. 62. 45. 67. 73. 71. 4027 P 158. 68. 48. 68. 79. 73. 4028 P 151. 57. 38. 67. 68. 69. 4029 P 158. 67. 44. 72. 87. 79. 4030 P 148. 68. 45. 68. 66. 62. 4031 P 138. 58. 46. 72. 80. 73. 4032 P 140. 58. 34. 65. 72. 75. 4033 P 184. 79. 41. 72. 100. 94. 4034 P 188. 77. 54. 85. 79. 74. 4035 P 158. 67. 46. 71. a 87. 4036 P 167. 70. 44. 76. 86. 90. 4037 P 134. 52. 39. 63. 65. 62. 4038 P 168. 72. 48. 82. 92. 97. 4039 P 159. 75. 51. 81. 92. 97. 4040 P 165. 74. 51. 60. 79. 86. 4041 P 159. 67. 48. 75. 84. 82. 4042 P 139. 68. 47. 80. 91. 98. 4043 P 131. 59. 44. 70. 74. 60. 4044 P 159. 75. 54. 74. 85. 96. 4045 P 131. 66. 45. 71. 78. 78. 4046 P 157. 71. 45. 69. 76. 83. 4047 P 163. 80. 59. 83. 94. 93. 4048 P 162. 73. 51. 76. 85. 88. 4049 P 143. 63. 44. 71. 84. 77. 4050 P 137. 62. 43. 66. 74. 70. ------------------------------------------------------------------------------------------------------------------------------------ P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAYS = DAYS OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. Spilled food precluded the calculation of this value.


Revised Draft Report Page 160 Testing Facility Study No. 20024504 PROTOCOL 20024504: AN EMBRYO-FETAL DEVELOPMENT STUDY OF THYMOSIN BETA 4 (FORMULATED AS RGN-532) BY INTRAVENOUS INJECTION IN RATS APPENDIX 6 (PAGE 3): MATERNAL FOOD CONSUMPTION VALUES - INDIVIDUAL DATA ------------------------------------------------------------------------------------------------------------------------------------ PREGNANCY STATUS DAYS 0 - 7 7 - 10 10 - 12 12 - 15 15 - 18 18 - 21 ------------------------------------------------------------------------------------------------------------------------------------ RAT # GROUP 3 RGN-352 60 MG/KG/DAY ------------------------------------------------------------------------------------------------------------------------------------ 4051 P 146. 60. 45. 73. 72. 83. 4052 P 146. 61. 46. 68. 80. 66. 4053 P 147. 60. 44. 68. 77. 70. 4054 P 185. 80. 56. 78. 80. 77. 4055 P 166. 71. 51. 79. 86. 77. 4056 P 146. 61. 47. 73. 74. 84. 4057 P 152. 63. 47. 62. 67. 93. 4058 P 141. 61. 42. 68. 69. 77. 4059 P 181. 81. 53. 86. 86. 92. 4060 P 137. 66. 54. 77. 86. 88. 4061 P 148. 67. 42. 77. 86. 88. 4062 P 173. 72. 55. 62. 84. 97. 4063 P 174. 75. 54. 80. 92. 99. 4064 P 135. 56. 42. 66. 78. 84. 4065 P 164. 70. 50. 81. 88. 90. 4066 P 172. 69. 56. 89. 104. 95. 4067 P 94. 60. 50. 73. 77. 86. 4068 P 146. 68. 45. 69. 78. 87. 4069 P 152. 63. 54. 80. 83. 93. 4070 P 161. 75. 53. 83. 80. 88. 4071 P 157. 68. 47. 74. 83. 91. 4072 P 165. 72. 50. 77. 83. 84. 4073 P 155. 75. 51. 74. 89. 89. 4074 P 155. 78. 42. 68. 93. 93. 4075 P 155. 70. 52. 72. 85. 93. ------------------------------------------------------------------------------------------------------------------------------------ P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAYS = DAYS OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G).


Revised Draft Report Page 161 Testing Facility Study No. 20024504 PROTOCOL 20024504: AN EMBRYO-FETAL DEVELOPMENT STUDY OF THYMOSIN BETA 4 (FORMULATED AS RGN-532) BY INTRAVENOUS INJECTION IN RATS APPENDIX 6 (PAGE 4): MATERNAL FOOD CONSUMPTION VALUES - INDIVIDUAL DATA ------------------------------------------------------------------------------------------------------------------------------------ PREGNANCY STATUS DAYS 0 - 7 7 - 10 10 - 12 12 - 15 15 - 18 18 - 21 ------------------------------------------------------------------------------------------------------------------------------------ RAT # GROUP 4 RGN-352 180 MG/KG/DAY ------------------------------------------------------------------------------------------------------------------------------------ 4076 P 156. 67. 48. 77. 82. 88. 4077 NP 110. 42. 32. 50. 49. 54. 4078 P 162. 70. 50. 73. 82. 80. 4079 P 169. 70. 47. 78. 89. 95. 4080 P 163. 70. 52. 74. 81. 67. 4081 P 150. 65. 44. 68. 82. 86. 4082 P 142. 62. 47. 77. 81. 90. 4083 P 156. 71. 48. 73. 78. 79. 4084 P 140. 67. 48. 76. 87. 88. 4085 P 149. 67. 46. 71. 86. 91. 4086 P 138. 66. 42. 69. 76. 83. 4087 P 154. 69. 51. 77. 84. 88. 4088 P 160. 76. 60. 97. 96. 96. 4089 P 147. 66. 46. 67. 81. 84. 4090 P 162. 73. 56. 83. 88. 87. 4091 P 166. 70. 57. 77. 83. 90. 4092 P 157. 71. 48. 74. 77. 87. 4093 P 138. 65. 41. 67. 82. 77. 4094 P 148. 72. 50. 81. 85. 85. 4095 P 170. 79. 51. 62. 80. 91. 4096 P 145. 66. 42. 65. 69. 83. 4097 P 152. 56. 45. 67. 80. 85. 4098 NP 142. 68. 43. 53. 58. 58. 4099 P 167. 72. 59. 84. 99. 101. 4100 P 156. 70. 46. 71. 88. 75. ------------------------------------------------------------------------------------------------------------------------------------ P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAYS = DAYS OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G).



Appendix 7 Necropsy Observations - Individual Data


Revised Draft Report Page 163 Testing Facility Study No. 20024504 PROTOCOL 20024504: AN EMBRYO-FETAL DEVELOPMENT STUDY OF THYMOSIN BETA 4 (FORMULATED AS RGN-532) BY INTRAVENOUS INJECTION IN RATS APPENDIX 7 (PAGE 1): NECROPSY OBSERVATIONS - INDIVIDUAL DATA ------------------------------------------------------------------------------------------------------------------------------------ RAT DAY OF PREGNANCY DOSES NUMBER NECROPSY STATUS ADMINISTERED OBSERVATIONS ------------------------------------------------------------------------------------------------------------------------------------ GROUP 1 CONTROL ARTICLE 0 (CONTROL) MG/KG/DAY ------------------------------------------------------------------------------------------------------------------------------------ 4001 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4002 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4003 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4004 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4005 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4006 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4007 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4008 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4009 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4010 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4011 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4012 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4013 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4014 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4015 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4016 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4017 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4018 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4019 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4020 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4021 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4022 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4023 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4024 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4025 DG 21 P 11 ALL TISSUES APPEARED NORMAL. ------------------------------------------------------------------------------------------------------------------------------------ DG = DAY OF PRESUMED GESTATION P = PREGNANT NP = NOT PREGNANT


Revised Draft Report Page 164 Testing Facility Study No. 20024504 PROTOCOL 20024504: AN EMBRYO-FETAL DEVELOPMENT STUDY OF THYMOSIN BETA 4 (FORMULATED AS RGN-532) BY INTRAVENOUS INJECTION IN RATS APPENDIX 7 (PAGE 2): NECROPSY OBSERVATIONS - INDIVIDUAL DATA ------------------------------------------------------------------------------------------------------------------------------------ RAT DAY OF PREGNANCY DOSES NUMBER NECROPSY STATUS ADMINISTERED OBSERVATIONS ------------------------------------------------------------------------------------------------------------------------------------ GROUP 2 RGN-352 20 MG/KG/DAY ------------------------------------------------------------------------------------------------------------------------------------ 4026 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4027 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4028 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4029 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4030 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4031 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4032 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4033 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4034 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4035 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4036 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4037 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4038 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4039 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4040 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4041 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4042 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4043 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4044 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4045 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4046 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4047 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4048 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4049 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4050 DG 21 P 11 ALL TISSUES APPEARED NORMAL. ------------------------------------------------------------------------------------------------------------------------------------ DG = DAY OF PRESUMED GESTATION P = PREGNANT NP = NOT PREGNANT


Revised Draft Report Page 165 Testing Facility Study No. 20024504 PROTOCOL 20024504: AN EMBRYO-FETAL DEVELOPMENT STUDY OF THYMOSIN BETA 4 (FORMULATED AS RGN-532) BY INTRAVENOUS INJECTION IN RATS APPENDIX 7 (PAGE 3): NECROPSY OBSERVATIONS - INDIVIDUAL DATA ------------------------------------------------------------------------------------------------------------------------------------ RAT DAY OF PREGNANCY DOSES NUMBER NECROPSY STATUS ADMINISTERED OBSERVATIONS ------------------------------------------------------------------------------------------------------------------------------------ GROUP 3 RGN-352 60 MG/KG/DAY ------------------------------------------------------------------------------------------------------------------------------------ 4051 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4052 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4053 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4054 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4055 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4056 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4057 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4058 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4059 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4060 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4061 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4062 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4063 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4064 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4065 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4066 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4067 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4068 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4069 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4070 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4071 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4072 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4073 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4074 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4075 DG 21 P 11 ALL TISSUES APPEARED NORMAL. ------------------------------------------------------------------------------------------------------------------------------------ DG = DAY OF PRESUMED GESTATION P = PREGNANT NP = NOT PREGNANT


Revised Draft Report Page 166 Testing Facility Study No. 20024504 PROTOCOL 20024504: AN EMBRYO-FETAL DEVELOPMENT STUDY OF THYMOSIN BETA 4 (FORMULATED AS RGN-532) BY INTRAVENOUS INJECTION IN RATS APPENDIX 7 (PAGE 4): NECROPSY OBSERVATIONS - INDIVIDUAL DATA ------------------------------------------------------------------------------------------------------------------------------------ RAT DAY OF PREGNANCY DOSES NUMBER NECROPSY STATUS ADMINISTERED OBSERVATIONS ------------------------------------------------------------------------------------------------------------------------------------ GROUP 4 RGN-352 180 MG/KG/DAY ------------------------------------------------------------------------------------------------------------------------------------ 4076 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4077 DG 21 NP 11 ALL TISSUES APPEARED NORMAL. 4078 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4079 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4080 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4081 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4082 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4083 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4084 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4085 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4086 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4087 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4088 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4089 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4090 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4091 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4092 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4093 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4094 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4095 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4096 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4097 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4098 DG 21 NP 11 ALL TISSUES APPEARED NORMAL. 4099 DG 21 P 11 ALL TISSUES APPEARED NORMAL. 4100 DG 21 P 11 ALL TISSUES APPEARED NORMAL. ------------------------------------------------------------------------------------------------------------------------------------ DG = DAY OF PRESUMED GESTATION P = PREGNANT NP = NOT PREGNANT



Appendix 8 Caesarean-Sectioning Observations - Individual Data


Revised Draft Report Page 168 Testing Facility Study No. 20024504 PROTOCOL 20024504: AN EMBRYO-FETAL DEVELOPMENT STUDY OF THYMOSIN BETA 4 (FORMULATED AS RGN-532) BY INTRAVENOUS INJECTION IN RATS APPENDIX 8 (PAGE 1): CAESAREAN-SECTIONING OBSERVATIONS - INDIVIDUAL DATA ------------------------------------------------------------------------------------------------------------------------------------ GROUP 1 CONTROL ARTICLE 0 (CONTROL) MG/KG/DAY ------------------------------------------------------------------------------------------------------------------------------------ VIABLE FETUSES DEAD FETUSES EARLY RESORPTIONS LATE RESORPTIONS IMPLANTATION SITES CORPORA LUTEA ------------------------------------------------------------------------------------------------------------------------------------ SEX RIGHT LEFT RIGHT LEFT RIGHT LEFT RIGHT LEFT RIGHT LEFT RIGHT LEFT RAT # M F HORN TOTAL HORN TOTAL HORN TOTAL HORN TOTAL HORN TOTAL OVARY TOTAL ------------------------------------------------------------------------------------------------------------------------------------ 4001 5 8 6 7 13 0 0 0 0 0 0 0 0 0 6 7 13 6 7 13 4002 8 4 6 6 12 0 0 0 0 0 0 0 0 0 6 6 12 6 6 12 4003 7 6 8 5 13 0 0 0 0 0 0 0 0 0 8 5 13 8 5 13 4004 5 12 9 8 17 0 0 0 1 0 1 0 0 0 10 8 18 10 8 18 4005 9 6 8 7 15 0 0 0 0 0 0 0 0 0 8 7 15 8 8 16 4006 6 9 8 7 15 0 0 0 0 0 0 0 0 0 8 7 15 10 7 17 4007 5 8 8 5 13 0 0 0 0 0 0 0 0 0 8 5 13 8 5 13 4008 5 8 5 8 13 0 0 0 0 0 0 0 0 0 5 8 13 5 8 13 4009 4 10 8 6 14 0 0 0 0 0 0 0 0 0 8 6 14 8 6 14 4010 8 6 6 8 14 0 0 0 1 1 2 0 1 1 7 10 17 7 10 17 4011 12 4 12 4 16 0 0 0 0 0 0 0 0 0 12 4 16 12 5 17 4012 3 8 6 5 11 0 0 0 1 1 2 0 0 0 7 6 13 7 6 13 4013 8 6 7 7 14 0 0 0 2 1 3 0 0 0 9 8 17 9 8 17 4014 3 8 6 5 11 0 0 0 2 2 4 0 0 0 8 7 15 8 8 16 4015 5 7 7 5 12 0 0 0 0 0 0 0 0 0 7 5 12 7 5 12 4016 9 4 6 7 13 0 0 0 0 0 0 0 0 0 6 7 13 7 7 14 4017 6 8 5 9 14 0 0 0 1 0 1 0 0 0 6 9 15 6 9 15 4018 8 8 8 8 16 0 0 0 0 0 0 0 0 0 8 8 16 8 8 16 4019 5 7 7 5 12 0 0 0 0 2 2 0 0 0 7 7 14 7 7 14 4020 6 6 7 5 12 0 0 0 1 0 1 0 0 0 8 5 13 8 5 13 4021 10 4 11 3 14 0 0 0 0 0 0 0 0 0 11 3 14 11 3 14 4022 5 10 5 10 15 0 0 0 2 2 4 0 0 0 7 12 19 7 12 19 4023 5 7 7 5 12 0 0 0 1 0 1 0 0 0 8 5 13 8 5 13 4024 4 7 6 5 11 0 0 0 1 3 4 0 0 0 7 8 15 7 8 15 4025 4 7 4 7 11 0 0 0 2 0 2 0 0 0 6 7 13 6 7 13 ------------------------------------------------------------------------------------------------------------------------------------ M = MALE F = FEMALE PLACENTAE APPEARED NORMAL UNLESS NOTED OTHERWISE.


Revised Draft Report Page 169 Testing Facility Study No. 20024504 PROTOCOL 20024504: AN EMBRYO-FETAL DEVELOPMENT STUDY OF THYMOSIN BETA 4 (FORMULATED AS RGN-532) BY INTRAVENOUS INJECTION IN RATS APPENDIX 8 (PAGE 2): CAESAREAN-SECTIONING OBSERVATIONS - INDIVIDUAL DATA ------------------------------------------------------------------------------------------------------------------------------------ GROUP 2 RGN-352 20 MG/KG/DAY ------------------------------------------------------------------------------------------------------------------------------------ VIABLE FETUSES DEAD FETUSES EARLY RESORPTIONS LATE RESORPTIONS IMPLANTATION SITES CORPORA LUTEA ------------------------------------------------------------------------------------------------------------------------------------ SEX RIGHT LEFT RIGHT LEFT RIGHT LEFT RIGHT LEFT RIGHT LEFT RIGHT LEFT RAT # M F HORN TOTAL HORN TOTAL HORN TOTAL HORN TOTAL HORN TOTAL OVARY TOTAL ------------------------------------------------------------------------------------------------------------------------------------ 4026 8 5 5 8 13 0 0 0 0 0 0 0 0 0 5 8 13 5 8 13 4027 10 4 9 5 14 0 0 0 0 0 0 0 0 0 9 5 14 9 5 14 4028 2 5 3 4 7 0 0 0 5 1 6 0 0 0 8 5 13 12 10 22 4029 6 7 5 8 13 0 0 0 0 0 0 0 0 0 5 8 13 5 9 14 4030 6 7 7 6 13 0 0 0 0 0 0 0 0 0 7 6 13 7 6 13 4031 6 8 8 6 14 0 0 0 0 1 1 0 0 0 8 7 15 8 8 16 4032 6 9 8 7 15 0 0 0 0 0 0 0 0 0 8 7 15 8 8 16 4033 10 6 9 7 16 0 0 0 0 0 0 0 0 0 9 7 16 9 7 16 4034 9 7 7 9 16 0 0 0 0 0 0 0 0 0 7 9 16 7 10 17 4035 3 11 6 8 14 0 0 0 1 0 1 0 0 0 7 8 15 7 8 15 4036 8 9 7 10 17 0 0 0 0 0 0 0 0 0 7 10 17 7 10 17 4037 7 7 6 8 14 0 0 0 0 0 0 0 0 0 6 8 14 6 8 14 4038 5 5 3 7 10 0 0 0 0 0 0 0 0 0 3 7 10 5 8 13 4039 4 8 2 10 12 0 0 0 1 1 2 0 0 0 3 11 14 4 11 15 4040 5 7 9 3 12 0 0 0 2 1 3 0 0 0 11 4 15 11 5 16 4041 8 8 8 8 16 0 0 0 0 0 0 0 0 0 8 8 16 8 9 17 4042 5 8 7 6 13 0 0 0 0 0 0 0 0 0 7 6 13 7 6 13 4043 6 5 5 6 11 0 0 0 0 0 0 0 0 0 5 6 11 5 6 11 4044 8 9 9 8 17 0 0 0 0 0 0 0 0 0 9 8 17 9 8 17 4045 5 6 5 6 11 0 0 0 0 1 1 0 0 0 5 7 12 5 7 12 4046 3 7 5 5 10 0 0 0 2 1 3 0 0 0 7 6 13 7 7 14 4047 8 7 10 5 15 0 0 0 0 0 0 0 0 0 10 5 15 10 6 16 4048 10 6 8 8 16 0 0 0 0 0 0 0 0 0 8 8 16 8 8 16 4049 6 10 11 5 16 0 0 0 0 0 0 0 0 0 11 5 16 11 5 16 4050 5 10 12 3 15 0 0 0 0 1 1 0 0 0 12 4 16 12 4 16 ------------------------------------------------------------------------------------------------------------------------------------ M = MALE F = FEMALE PLACENTAE APPEARED NORMAL UNLESS NOTED OTHERWISE.


Revised Draft Report Page 170 Testing Facility Study No. 20024504 PROTOCOL 20024504: AN EMBRYO-FETAL DEVELOPMENT STUDY OF THYMOSIN BETA 4 (FORMULATED AS RGN-532) BY INTRAVENOUS INJECTION IN RATS APPENDIX 8 (PAGE 3): CAESAREAN-SECTIONING OBSERVATIONS - INDIVIDUAL DATA ------------------------------------------------------------------------------------------------------------------------------------ GROUP 3 RGN-352 60 MG/KG/DAY ------------------------------------------------------------------------------------------------------------------------------------ VIABLE FETUSES DEAD FETUSES EARLY RESORPTIONS LATE RESORPTIONS IMPLANTATION SITES CORPORA LUTEA ------------------------------------------------------------------------------------------------------------------------------------ SEX RIGHT LEFT RIGHT LEFT RIGHT LEFT RIGHT LEFT RIGHT LEFT RIGHT LEFT RAT # M F HORN TOTAL HORN TOTAL HORN TOTAL HORN TOTAL HORN TOTAL OVARY TOTAL ------------------------------------------------------------------------------------------------------------------------------------ 4051 4 9 4 9 13 0 0 0 0 0 0 0 0 0 4 9 13 4 9 13 4052 7 8 10 5 15 0 0 0 1 0 1 0 0 0 11 5 16 11 5 16 4053 4 8 4 8 12 0 0 0 0 1 1 0 0 0 4 9 13 4 9 13 4054 8 4 6 6 12 0 0 0 0 0 0 0 0 0 6 6 12 6 6 12 4055 8 7 7 8 15 0 0 0 1 0 1 0 0 0 8 8 16 8 8 16 4056 5 6 6 5 11 0 0 0 1 0 1 0 0 0 7 5 12 7 5 12 4057 5 7 3 9 12 0 0 0 1 0 1 0 0 0 4 9 13 4 9 13 4058 6 7 7 6 13 0 0 0 0 0 0 0 0 0 7 6 13 7 6 13 4059 8 3 3 8 11 0 0 0 1 0 1 0 0 0 4 8 12 4 8 12 4060 8 6 7 7 14 0 0 0 0 1 1 0 0 0 7 8 15 8 9 17 4061 4 11 9 6 15 0 0 0 1 0 1 0 0 0 10 6 16 10 7 17 4062 3 12 10 5 15 0 0 0 0 0 0 0 0 0 10 5 15 11 5 16 4063 11 5 10 6 16 0 0 0 0 0 0 0 0 0 10 6 16 10 6 16 4064 9 4 8 5 13 0 0 0 0 2 2 0 0 0 8 7 15 8 8 16 4065 12 2 9 5 14 0 0 0 0 0 0 0 0 0 9 5 14 9 5 14 4066 6 7 9 4 13 0 0 0 1 0 1 0 0 0 10 4 14 10 5 15 4067 4 11 8 7 15 0 0 0 0 0 0 0 0 0 8 7 15 8 8 16 4068 10 3 6 7 13 0 0 0 0 0 0 0 0 0 6 7 13 7 7 14 4069 8 6 9 5 14 0 0 0 0 0 0 0 0 0 9 5 14 9 5 14 4070 10 5 8 7 15 0 0 0 1 0 1 0 0 0 9 7 16 9 7 16 4071 7 8 7 8 15 0 0 0 0 0 0 0 0 0 7 8 15 7 8 15 4072 11 5 9 7 16 0 0 0 1 0 1 0 0 0 10 7 17 10 7 17 4073 8 5 6 7 13 0 0 0 1 1 2 0 0 0 7 8 15 7 8 15 4074 5 3 8 0 8 0 0 0 0 0 0 0 0 0 8 0 8 8 2 10 4075 6 9 9 6 15 0 0 0 0 0 0 0 0 0 9 6 15 10 6 16 ------------------------------------------------------------------------------------------------------------------------------------ M = MALE F = FEMALE PLACENTAE APPEARED NORMAL UNLESS NOTED OTHERWISE.


Revised Draft Report Page 171 Testing Facility Study No. 20024504 PROTOCOL 20024504: AN EMBRYO-FETAL DEVELOPMENT STUDY OF THYMOSIN BETA 4 (FORMULATED AS RGN-532) BY INTRAVENOUS INJECTION IN RATS APPENDIX 8 (PAGE 4): CAESAREAN-SECTIONING OBSERVATIONS - INDIVIDUAL DATA ------------------------------------------------------------------------------------------------------------------------------------ GROUP 4 RGN-352 180 MG/KG/DAY ------------------------------------------------------------------------------------------------------------------------------------ VIABLE FETUSES DEAD FETUSES EARLY RESORPTIONS LATE RESORPTIONS IMPLANTATION SITES CORPORA LUTEA ------------------------------------------------------------------------------------------------------------------------------------ SEX RIGHT LEFT RIGHT LEFT RIGHT LEFT RIGHT LEFT RIGHT LEFT RIGHT LEFT RAT # M F HORN TOTAL HORN TOTAL HORN TOTAL HORN TOTAL HORN TOTAL OVARY TOTAL ------------------------------------------------------------------------------------------------------------------------------------ 4076 6 6 6 6 12 0 0 0 0 1 1 0 0 0 6 7 13 6 8 14 4077 NOT PREGNANT 4078 6 6 7 5 12 0 0 0 0 0 0 0 0 0 7 5 12 7 5 12 4079 4 8 7 5 12 0 0 0 1 0 1 0 0 0 8 5 13 8 5 13 4080 6 6 9 3 12 0 0 0 1 0 1 0 0 0 10 3 13 10 4 14 4081 10 4 8 6 14 0 0 0 0 0 0 0 0 0 8 6 14 8 6 14 4082 10 6 6 10 16 0 0 0 0 0 0 0 0 0 6 10 16 6 10 16 4083 4 12 6 10 16 0 0 0 0 0 0 0 0 0 6 10 16 6 10 16 4084 7 9 8 8 16 0 0 0 0 0 0 0 0 0 8 8 16 8 8 16 4085 8 5 6 7 13 0 0 0 1 0 1 0 0 0 7 7 14 7 7 14 4086 6 5 9 2 11 0 0 0 0 0 0 0 0 0 9 2 11 9 3 12 4087 7 7 7 7 14 0 0 0 0 0 0 0 0 0 7 7 14 7 7 14 4088 5 10 6 9 15 0 0 0 0 0 0 0 0 0 6 9 15 6 10 16 4089 8 5 8 5 13 0 0 0 0 0 0 0 0 0 8 5 13 8 5 13 4090 8 6 7 7 14 0 0 0 0 0 0 0 0 0 7 7 14 7 7 14 4091 9 5 6 8 14 0 0 0 0 0 0 0 0 0 6 8 14 7 8 15 4092 4 9 9 4 13 0 0 0 0 0 0 0 0 0 9 4 13 9 4 13 4093 8 5 8 5 13 0 0 0 2 0 2 0 0 0 10 5 15 10 6 16 4094 9 7 8 8 16 0 0 0 0 1 1 0 0 0 8 9 17 8 9 17 4095 7 10 7 10 17 0 0 0 0 0 0 0 0 0 7 10 17 7 10 17 4096 10 6 9 7 16 0 0 0 1 0 1 0 0 0 10 7 17 10 7 17 4097 8 6 8 6 14 0 0 0 0 0 0 0 0 0 8 6 14 8 6 14 4098 NOT PREGNANT 4099 6 10 8 8 16 0 0 0 0 0 0 0 0 0 8 8 16 8 8 16 4100 10 5 4 11 15 0 0 0 3 0 3 0 0 0 7 11 18 7 12 19 ------------------------------------------------------------------------------------------------------------------------------------ M = MALE F = FEMALE PLACENTAE APPEARED NORMAL UNLESS NOTED OTHERWISE.



Appendix 9 Litter Observations (Caesarean-Delivered Fetuses) - Individual Data


Revised Draft Report Page 173 Testing Facility Study No. 20024504 PROTOCOL 20024504: AN EMBRYO-FETAL DEVELOPMENT STUDY OF THYMOSIN BETA 4 (FORMULATED AS RGN-532) BY INTRAVENOUS INJECTION IN RATS APPENDIX 9 (PAGE 1): LITTER OBSERVATIONS (CAESAREAN-DELIVERED FETUSES) - INDIVIDUAL DATA ------------------------------------------------------------------------------------------------------------------------------------ GROUP 1 CONTROL ARTICLE 0 (CONTROL) MG/KG/DAY ------------------------------------------------------------------------------------------------------------------------------------ NUMBER OF LIVE AVERAGE FETAL ------ CONCEPTUSES ------ FETUSES BODY WEIGHT (G) TOTAL RESORBED ------------------------------------------------------------------------------------------------------------------------------------ RAT # MALE FEMALE TOTAL MALE FEMALE TOTAL a N N % ------------------------------------------------------------------------------------------------------------------------------------ 4001 5 8 13 5.60 5.39 5.47 13 0 0.0 4002 8 4 12 5.68 5.41 5.59 12 0 0.0 4003 7 6 13 6.20 5.81 6.02 13 0 0.0 4004 5 12 17 5.91 5.72 5.78 18 1 5.6 4005 9 6 15 5.74 5.25 5.54 15 0 0.0 4006 6 9 15 5.69 5.63 5.65 15 0 0.0 4007 5 8 13 5.88 5.67 5.75 13 0 0.0 4008 5 8 13 5.93 5.59 5.72 13 0 0.0 4009 4 10 14 5.32 5.45 5.41 14 0 0.0 4010 8 6 14 5.12 4.54 4.87 17 3 17.6 4011 12 4 16 5.29 5.18 5.27 16 0 0.0 4012 3 8 11 6.26 5.91 6.01 13 2 15.4 4013 8 6 14 5.73 5.35 5.56 17 3 17.6 4014 3 8 11 6.17 5.84 5.93 15 4 26.7 4015 5 7 12 5.70 5.50 5.58 12 0 0.0 4016 9 4 13 5.81 5.45 5.70 13 0 0.0 4017 6 8 14 6.17 5.82 5.97 15 1 6.7 4018 8 8 16 5.17 5.06 5.11 16 0 0.0 4019 5 7 12 4.79 4.81 4.80 14 2 14.3 4020 6 6 12 5.64 5.31 5.48 13 1 7.7 4021 10 4 14 5.70 5.76 5.72 14 0 0.0 4022 5 10 15 5.74 5.29 5.44 19 4 21.0 4023 5 7 12 5.38 5.21 5.28 13 1 7.7 4024 4 7 11 5.81 5.45 5.58 15 4 26.7 4025 4 7 11 5.38 5.23 5.28 13 2 15.4 ------------------------------------------------------------------------------------------------------------------------------------ a. TOTAL = SUM OF FETAL WEIGHTS/NUMBER OF LIVE FETUSES.


Revised Draft Report Page 174 Testing Facility Study No. 20024504 PROTOCOL 20024504: AN EMBRYO-FETAL DEVELOPMENT STUDY OF THYMOSIN BETA 4 (FORMULATED AS RGN-532) BY INTRAVENOUS INJECTION IN RATS APPENDIX 9 (PAGE 2): LITTER OBSERVATIONS (CAESAREAN-DELIVERED FETUSES) - INDIVIDUAL DATA ------------------------------------------------------------------------------------------------------------------------------------ GROUP 2 RGN-352 20 MG/KG/DAY ------------------------------------------------------------------------------------------------------------------------------------ NUMBER OF LIVE AVERAGE FETAL ------ CONCEPTUSES ------ FETUSES BODY WEIGHT (G) TOTAL RESORBED ------------------------------------------------------------------------------------------------------------------------------------ RAT # MALE FEMALE TOTAL MALE FEMALE TOTAL a N N % ------------------------------------------------------------------------------------------------------------------------------------ 4026 8 5 13 5.59 5.59 5.59 13 0 0.0 4027 10 4 14 5.64 5.04 5.47 14 0 0.0 4028 2 5 7 5.72 4.86 5.10 13 6 46.2 4029 6 7 13 6.10 5.64 5.85 13 0 0.0 4030 6 7 13 5.90 5.88 5.89 13 0 0.0 4031 6 8 14 5.98 5.60 5.76 15 1 6.7 4032 6 9 15 6.00 5.74 5.84 15 0 0.0 4033 10 6 16 6.28 6.11 6.22 16 0 0.0 4034 9 7 16 5.62 5.11 5.39 16 0 0.0 4035 3 11 14 5.76 5.60 5.63 15 1 6.7 4036 8 9 17 5.89 5.64 5.76 17 0 0.0 4037 7 7 14 5.54 5.26 5.40 14 0 0.0 4038 5 5 10 6.06 5.77 5.92 10 0 0.0 4039 4 8 12 6.04 5.68 5.80 14 2 14.3 4040 5 7 12 5.61 5.64 5.63 15 3 20.0 4041 8 8 16 5.88 5.61 5.74 16 0 0.0 4042 5 8 13 5.61 5.53 5.56 13 0 0.0 4043 6 5 11 4.60 5.23 4.88 11 0 0.0 4044 8 9 17 5.43 5.30 5.36 17 0 0.0 4045 5 6 11 5.00 5.75 5.41 12 1 8.3 4046 3 7 10 6.26 5.20 5.52 13 3 23.1 4047 8 7 15 6.00 5.69 5.86 15 0 0.0 4048 10 6 16 5.61 5.43 5.54 16 0 0.0 4049 6 10 16 5.55 5.32 5.41 16 0 0.0 4050 5 10 15 5.36 5.13 5.20 16 1 6.2 ------------------------------------------------------------------------------------------------------------------------------------ a. TOTAL = SUM OF FETAL WEIGHTS/NUMBER OF LIVE FETUSES.


Revised Draft Report Page 175 Testing Facility Study No. 20024504 PROTOCOL 20024504: AN EMBRYO-FETAL DEVELOPMENT STUDY OF THYMOSIN BETA 4 (FORMULATED AS RGN-532) BY INTRAVENOUS INJECTION IN RATS APPENDIX 9 (PAGE 3): LITTER OBSERVATIONS (CAESAREAN-DELIVERED FETUSES) - INDIVIDUAL DATA ------------------------------------------------------------------------------------------------------------------------------------ GROUP 3 RGN-352 60 MG/KG/DAY ------------------------------------------------------------------------------------------------------------------------------------ NUMBER OF LIVE AVERAGE FETAL ------ CONCEPTUSES ------ FETUSES BODY WEIGHT (G) TOTAL RESORBED ------------------------------------------------------------------------------------------------------------------------------------ RAT # MALE FEMALE TOTAL MALE FEMALE TOTAL a N N % ------------------------------------------------------------------------------------------------------------------------------------ 4051 4 9 13 5.74 5.40 5.50 13 0 0.0 4052 7 8 15 5.51 5.33 5.41 16 1 6.2 4053 4 8 12 6.00 5.77 5.84 13 1 7.7 4054 8 4 12 6.16 5.82 6.05 12 0 0.0 4055 8 7 15 5.65 5.19 5.44 16 1 6.2 4056 5 6 11 5.98 5.72 5.84 12 1 8.3 4057 5 7 12 6.39 5.68 5.98 13 1 7.7 4058 6 7 13 5.32 5.11 5.21 13 0 0.0 4059 8 3 11 5.91 5.89 5.90 12 1 8.3 4060 8 6 14 5.99 5.80 5.91 15 1 6.7 4061 4 11 15 5.98 5.45 5.59 16 1 6.2 4062 3 12 15 5.64 5.63 5.63 15 0 0.0 4063 11 5 16 6.11 5.79 6.01 16 0 0.0 4064 9 4 13 5.88 5.60 5.80 15 2 13.3 4065 12 2 14 5.75 5.72 5.75 14 0 0.0 4066 6 7 13 6.60 6.10 6.33 14 1 7.1 4067 4 11 15 5.86 5.70 5.74 15 0 0.0 4068 10 3 13 5.74 5.39 5.66 13 0 0.0 4069 8 6 14 6.05 5.72 5.91 14 0 0.0 4070 10 5 15 5.74 5.39 5.62 16 1 6.2 4071 7 8 15 6.37 6.05 6.20 15 0 0.0 4072 11 5 16 5.56 5.26 5.46 17 1 5.9 4073 8 5 13 6.04 5.70 5.91 15 2 13.3 4074 5 3 8 6.33 6.20 6.28 8 0 0.0 4075 6 9 15 5.84 5.61 5.70 15 0 0.0 ------------------------------------------------------------------------------------------------------------------------------------ a. TOTAL = SUM OF FETAL WEIGHTS/NUMBER OF LIVE FETUSES.


Revised Draft Report Page 176 Testing Facility Study No. 20024504 PROTOCOL 20024504: AN EMBRYO-FETAL DEVELOPMENT STUDY OF THYMOSIN BETA 4 (FORMULATED AS RGN-532) BY INTRAVENOUS INJECTION IN RATS APPENDIX 9 (PAGE 4): LITTER OBSERVATIONS (CAESAREAN-DELIVERED FETUSES) - INDIVIDUAL DATA ------------------------------------------------------------------------------------------------------------------------------------ GROUP 4 RGN-352 180 MG/KG/DAY ------------------------------------------------------------------------------------------------------------------------------------ NUMBER OF LIVE AVERAGE FETAL ------ CONCEPTUSES ------ FETUSES BODY WEIGHT (G) TOTAL RESORBED ------------------------------------------------------------------------------------------------------------------------------------ RAT # MALE FEMALE TOTAL MALE FEMALE TOTAL a N N % ------------------------------------------------------------------------------------------------------------------------------------ 4076 6 6 12 5.79 5.38 5.59 13 1 7.7 4077 NOT PREGNANT 4078 6 6 12 6.02 5.84 5.93 12 0 0.0 4079 4 8 12 5.71 5.73 5.72 13 1 7.7 4080 6 6 12 5.58 5.48 5.53 13 1 7.7 4081 10 4 14 5.70 5.23 5.57 14 0 0.0 4082 10 6 16 5.77 5.52 5.68 16 0 0.0 4083 4 12 16 5.14 5.09 5.10 16 0 0.0 4084 7 9 16 5.47 5.13 5.28 16 0 0.0 4085 8 5 13 6.20 6.03 6.13 14 1 7.1 4086 6 5 11 6.14 5.87 6.02 11 0 0.0 4087 7 7 14 6.19 5.59 5.89 14 0 0.0 4088 5 10 15 6.02 5.84 5.90 15 0 0.0 4089 8 5 13 6.37 5.97 6.22 13 0 0.0 4090 8 6 14 5.70 5.24 5.50 14 0 0.0 4091 9 5 14 6.04 5.80 5.96 14 0 0.0 4092 4 9 13 5.15 5.09 5.11 13 0 0.0 4093 8 5 13 6.13 5.84 6.02 15 2 13.3 4094 9 7 16 5.79 5.78 5.79 17 1 5.9 4095 7 10 17 5.86 5.55 5.68 17 0 0.0 4096 10 6 16 5.67 5.15 5.48 17 1 5.9 4097 8 6 14 5.71 5.02 5.41 14 0 0.0 4098 NOT PREGNANT 4099 6 10 16 6.20 5.82 5.96 16 0 0.0 4100 10 5 15 6.04 5.47 5.85 18 3 16.7 ------------------------------------------------------------------------------------------------------------------------------------ a. TOTAL = SUM OF FETAL WEIGHTS/NUMBER OF LIVE FETUSES.



Appendix 10 Fetal Sex, Vital Status and Body Weight - Individual Data


Revised Draft Report Page 178 Testing Facility Study No. 20024504 PROTOCOL 20024504: AN EMBRYO-FETAL DEVELOPMENT STUDY OF THYMOSIN BETA 4 (FORMULATED AS RGN-532) BY INTRAVENOUS INJECTION IN RATS APPENDIX 10 (PAGE 1): FETAL SEX, VITAL STATUS AND BODY WEIGHT - INDIVIDUAL DATA ------------------------------------------------------------------------------------------------------------------------------------ GROUP 1 CONTROL ARTICLE 0 (CONTROL) MG/KG/DAY ------------------------------------------------------------------------------------------------------------------------------------ FETUS # 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 ------------------------------------------------------------------------------------------------------------------------------------ RAT # CLs 4001 6/ 7 FA FA FA FA FA FA / FA MA MA FA MA MA MA 4.81 5.25 5.68 5.75 5.30 5.41 5.50 5.67 5.57 5.39 5.53 5.50 5.71 4002 6/ 6 FA MA MA MA FA MA / MA FA MA FA MA MA 5.52 5.53 5.66 5.64 5.61 5.69 5.84 5.68 6.04 4.82 5.47 5.60 4003 8/ 5 FA MA MA FA FA FA FA MA / MA FA MA MA MA 5.95 6.16 6.45 5.55 6.25 5.72 5.43 6.40 5.81 5.94 6.19 6.28 6.10 4004 10/ 8 MA FA FA FA MA FA FA FA FA E / FA MA MA FA MA FA FA FA 6.02 5.86 5.87 5.72 5.84 5.58 5.65 5.85 6.01 5.68 5.99 5.91 5.31 5.81 5.52 5.64 5.92 4005 8/ 8 MA MA MA MA FA MA MA MA / FA FA FA FA MA MA FA 5.66 5.66 5.50 6.15 5.39 5.64 5.72 5.54 5.19 5.50 5.56 4.95 5.59 6.17 4.92 4006 10/ 7 FA MA FA FA MA FA FA MA / FA MA FA MA MA FA FA 5.00 5.51 5.62 5.57 5.67 5.73 5.64 5.65 5.30 5.66 5.98 5.67 5.96 6.08 5.78 4007 8/ 5 FA MA FA FA MA FA MA FA / FA FA MA FA MA 5.49 6.05 5.85 5.55 5.85 5.77 5.92 5.84 5.44 5.37 5.81 6.08 5.78 4008 5/ 8 FA FA MA FA FA / MA FA FA MA FA MA FA MA 5.52 6.12 5.90 5.42 5.58 5.96 5.59 5.37 5.85 5.36 6.18 5.75 5.74 4009 8/ 6 MA FA FA MA FA FA FA FA / FA MA FA FA MA FA 5.59 5.38 5.45 4.16 5.70 5.63 5.33 5.23 5.43 5.67 5.65 5.29 5.84 5.42 4010 7/10 FA E MA MA MA FA MA / FA FA FA MA FA MA MA E L MA 3.08 3.72 6.05 6.06 6.03 5.90 4.67 3.79 4.96 5.89 4.73 6.30 4.14 0.15 2.88 4011 12/ 5 MA MA MA FA FA FA MA MA MA MA MA MA / FA MA MA MA 5.22 5.36 5.34 5.20 5.20 5.20 5.43 5.36 5.06 5.42 5.33 5.77 5.14 5.56 4.15 5.52 4012 7/ 6 FA FA FA FA E FA FA / FA MA MA MA FA E 5.82 5.56 5.92 5.65 6.51 6.06 5.87 6.30 6.27 6.22 5.89 4013 9/ 8 MA FA MA MA MA MA E E FA / MA E FA MA FA FA FA MA 5.57 5.62 5.65 5.87 5.41 5.77 4.70 5.81 5.45 6.00 5.21 5.36 5.75 5.74 4014 8/ 8 FA FA E E FA MA FA FA / E E FA FA FA MA MA 5.77 5.66 5.71 5.81 5.75 6.03 5.81 5.63 6.32 6.34 6.37 4015 7/ 5 FA MA FA MA MA MA FA / FA FA FA MA FA 5.68 5.73 4.22 6.25 6.26 5.98 5.71 5.81 5.51 5.57 4.26 6.03 4016 7/ 7 FA FA FA MA MA MA / MA FA MA MA MA MA MA 5.33 5.67 5.35 5.89 5.88 6.10 5.79 5.46 5.55 5.29 5.85 6.18 5.74 4017 6/ 9 FA FA FA MA E MA / MA FA MA MA FA FA MA FA FA 5.75 6.23 5.61 6.19 5.97 6.48 5.85 6.02 6.03 6.02 5.82 6.31 5.73 5.58 4018 8/ 8 MA MA FA MA MA MA FA FA / FA MA FA MA MA FA FA FA 5.33 4.78 5.20 5.25 5.10 5.36 5.03 4.95 4.73 5.26 5.32 5.26 5.00 5.05 4.98 5.20 ------------------------------------------------------------------------------------------------------------------------------------ M = MALE F = FEMALE A = ALIVE E = EARLY RESORPTION L = LATE RESORPTION "/" DENOTES POSITION OF CERVIX CLs = CORPORA LUTEA/OVARY FETAL BODY WEIGHTS WERE RECORDED IN GRAMS (G).


Revised Draft Report Page 179 Testing Facility Study No. 20024504 PROTOCOL 20024504: AN EMBRYO-FETAL DEVELOPMENT STUDY OF THYMOSIN BETA 4 (FORMULATED AS RGN-532) BY INTRAVENOUS INJECTION IN RATS APPENDIX 10 (PAGE 2): FETAL SEX, VITAL STATUS AND BODY WEIGHT - INDIVIDUAL DATA ------------------------------------------------------------------------------------------------------------------------------------ GROUP 1 CONTROL ARTICLE 0 (CONTROL) MG/KG/DAY ------------------------------------------------------------------------------------------------------------------------------------ FETUS # 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 ------------------------------------------------------------------------------------------------------------------------------------ RAT # CLs 4019 7/ 7 FA MA FA FA FA MA FA / MA FA E E FA MA MA 4.15 2.93 5.01 5.52 3.73 6.07 5.40 3.03 5.70 4.18 6.01 5.89 4020 8/ 5 MA MA FA MA FA FA MA E / MA MA FA FA FA 5.07 5.46 5.10 5.65 5.25 5.31 6.20 5.54 5.90 5.41 5.61 5.20 4021 11/ 3 MA FA MA MA MA FA MA MA MA MA FA / MA MA FA 5.23 5.93 5.73 5.54 5.50 5.85 5.94 5.31 5.62 5.96 5.57 6.03 6.17 5.69 4022 7/12 FA E FA E FA FA FA / FA FA MA FA E FA MA MA MA FA MA E 5.51 5.78 5.68 5.66 5.91 5.51 4.86 5.94 4.85 3.60 5.63 5.50 5.69 5.55 5.96 4023 8/ 5 FA FA FA E MA MA FA FA / MA MA FA FA MA 4.68 5.49 5.20 5.49 5.25 5.06 5.26 5.26 5.24 5.75 5.05 5.68 4024 7/ 8 FA MA FA FA FA E MA / MA FA FA E MA E FA E 5.42 5.76 5.63 5.83 5.55 5.99 5.60 5.31 5.33 5.90 5.11 4025 6/ 7 MA FA FA E FA E / FA MA MA FA FA MA FA 5.32 5.66 5.08 5.07 5.07 5.31 5.54 5.09 5.28 5.35 5.35 ------------------------------------------------------------------------------------------------------------------------------------ M = MALE F = FEMALE A = ALIVE E = EARLY RESORPTION L = LATE RESORPTION "/" DENOTES POSITION OF CERVIX CLs = CORPORA LUTEA/OVARY FETAL BODY WEIGHTS WERE RECORDED IN GRAMS (G).


Revised Draft Report Page 180 Testing Facility Study No. 20024504 PROTOCOL 20024504: AN EMBRYO-FETAL DEVELOPMENT STUDY OF THYMOSIN BETA 4 (FORMULATED AS RGN-532) BY INTRAVENOUS INJECTION IN RATS APPENDIX 10 (PAGE 3): FETAL SEX, VITAL STATUS AND BODY WEIGHT - INDIVIDUAL DATA ------------------------------------------------------------------------------------------------------------------------------------ GROUP 2 RGN-352 20 MG/KG/DAY ------------------------------------------------------------------------------------------------------------------------------------ FETUS # 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 ------------------------------------------------------------------------------------------------------------------------------------ RAT # CLs 4026 5/ 8 FA FA FA FA MA / MA MA MA MA MA MA MA FA 5.22 5.43 5.81 5.60 5.67 5.64 5.54 5.61 5.67 5.31 5.58 5.67 5.88 4027 9/ 5 FA FA MA MA MA FA FA MA MA / MA MA MA MA MA 5.10 5.18 5.67 5.09 5.49 5.01 4.86 5.63 5.56 5.40 5.72 6.10 5.71 6.05 4028 12/10 E E FA E FA E MA E / FA FA MA E FA 4.06 5.22 5.84 4.41 5.34 5.59 5.26 4029 5/ 9 FA MA FA FA FA / FA MA FA MA MA MA MA FA 5.84 6.25 5.85 5.79 5.88 5.42 5.96 5.25 6.05 5.92 6.22 6.18 5.47 4030 7/ 6 FA FA MA MA FA FA FA / MA MA MA MA FA FA 5.80 5.79 6.00 6.18 5.95 5.97 6.04 6.13 5.53 5.65 5.90 5.97 5.68 4031 8/ 8 FA MA FA FA MA FA FA FA / FA E MA MA MA MA FA 5.85 5.72 5.90 5.47 6.07 5.09 5.88 5.22 5.73 6.25 6.11 5.82 5.88 5.65 4032 8/ 8 MA FA FA FA MA MA MA MA / FA FA FA FA FA FA MA 5.78 6.00 5.20 5.67 6.37 5.75 5.83 5.86 5.71 5.85 6.20 5.72 5.58 5.71 6.40 4033 9/ 7 FA MA MA MA MA FA FA FA MA / MA MA MA FA FA MA MA 5.95 6.34 5.98 6.19 6.36 6.20 6.47 6.20 6.45 6.10 6.50 6.31 6.04 5.82 6.32 6.28 4034 7/10 MA FA MA MA MA FA MA / MA FA MA MA FA FA FA FA MA 5.29 4.72 5.73 5.70 5.97 5.23 5.72 5.52 5.12 5.15 5.70 4.95 5.34 5.04 5.35 5.78 4035 7/ 8 FA FA FA MA E FA MA / MA FA FA FA FA FA FA FA 5.92 5.71 5.56 5.74 5.78 5.71 5.84 5.58 5.20 5.76 5.64 5.52 5.48 5.40 4036 7/10 MA FA FA FA FA MA FA / FA FA MA MA MA MA MA FA MA FA 6.18 5.36 5.32 5.66 5.54 5.76 5.97 5.48 5.72 5.94 5.77 5.88 5.90 6.02 5.77 5.68 5.98 4037 6/ 8 MA FA MA MA FA FA / MA MA MA MA FA FA FA FA 5.05 5.46 5.73 5.75 5.45 5.04 5.21 5.61 6.01 5.43 5.64 4.98 5.01 5.28 4038 5/ 8 FA MA MA / FA FA MA MA FA FA MA 5.99 6.55 6.18 5.75 5.56 6.23 5.70 5.65 5.90 5.66 4039 4/11 MA E MA / E MA FA FA FA FA MA FA FA FA FA 6.33 6.34 5.61 5.62 5.05 5.44 6.05 5.87 5.84 5.90 5.79 5.75 4040 11/ 5 MA MA FA FA FA E MA FA FA MA E / FA FA MA E 4.37 5.34 5.59 5.27 5.87 6.12 5.73 5.55 5.97 5.87 5.63 6.27 4041 8/ 9 FA FA FA MA FA MA MA FA / MA MA FA FA MA MA MA FA 5.38 5.83 5.50 5.90 5.58 5.83 5.91 5.43 6.33 5.89 5.57 5.73 5.96 5.37 5.81 5.83 4042 7/ 6 FA FA FA FA FA FA FA / MA MA FA MA MA MA 5.51 5.62 5.35 5.90 5.61 5.51 5.42 5.04 5.77 5.35 5.88 5.92 5.42 4043 5/ 6 FA MA MA MA MA / MA FA FA MA FA FA 4.99 5.54 2.47 5.28 3.32 5.31 5.47 5.27 5.68 5.19 5.21 ------------------------------------------------------------------------------------------------------------------------------------ M = MALE F = FEMALE A = ALIVE E = EARLY RESORPTION L = LATE RESORPTION "/" DENOTES POSITION OF CERVIX CLs = CORPORA LUTEA/OVARY FETAL BODY WEIGHTS WERE RECORDED IN GRAMS (G).


Revised Draft Report Page 181 Testing Facility Study No. 20024504 PROTOCOL 20024504: AN EMBRYO-FETAL DEVELOPMENT STUDY OF THYMOSIN BETA 4 (FORMULATED AS RGN-532) BY INTRAVENOUS INJECTION IN RATS APPENDIX 10 (PAGE 4): FETAL SEX, VITAL STATUS AND BODY WEIGHT - INDIVIDUAL DATA ------------------------------------------------------------------------------------------------------------------------------------ GROUP 2 RGN-352 20 MG/KG/DAY ------------------------------------------------------------------------------------------------------------------------------------ FETUS # 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 ------------------------------------------------------------------------------------------------------------------------------------ RAT # CLs 4044 9/ 8 FA MA FA FA MA FA MA FA FA / FA MA FA MA MA FA MA MA 5.11 5.09 5.28 5.63 5.69 5.02 5.73 5.23 5.61 5.26 5.63 5.27 5.31 5.60 5.29 6.03 4.38 4045 5/ 7 MA MA MA MA FA / MA FA FA FA FA E FA 3.58 6.42 6.23 2.37 5.99 6.39 4.70 6.17 5.89 5.76 5.97 4046 7/ 7 E MA FA MA FA FA E / FA FA FA FA E MA 6.18 3.50 6.14 5.82 5.63 5.51 4.14 5.77 6.03 6.46 4047 10/ 6 FA MA MA MA MA FA FA MA FA MA / FA FA FA MA MA 5.40 5.84 5.53 5.81 6.20 6.22 5.41 6.07 5.60 5.98 5.41 6.01 5.79 6.13 6.44 4048 8/ 8 FA FA FA MA FA MA FA MA / FA MA MA MA MA MA MA MA 5.10 5.36 5.57 5.56 5.42 5.50 5.40 5.44 5.71 6.15 5.91 5.69 5.56 5.85 4.84 5.64 4049 11/ 5 MA FA FA MA FA MA MA FA FA MA MA / FA FA FA FA FA 5.35 5.21 5.22 5.81 4.95 5.72 4.96 5.04 5.11 5.43 6.03 5.30 5.51 5.58 5.51 5.80 4050 12/ 4 FA MA FA FA FA MA MA FA MA FA FA MA / FA FA E FA 4.82 5.36 5.21 5.24 5.10 5.39 5.55 4.97 5.49 5.08 4.98 4.99 5.44 4.94 5.50 ------------------------------------------------------------------------------------------------------------------------------------ M = MALE F = FEMALE A = ALIVE E = EARLY RESORPTION L = LATE RESORPTION "/" DENOTES POSITION OF CERVIX CLs = CORPORA LUTEA/OVARY FETAL BODY WEIGHTS WERE RECORDED IN GRAMS (G).


Revised Draft Report Page 182 Testing Facility Study No. 20024504 PROTOCOL 20024504: AN EMBRYO-FETAL DEVELOPMENT STUDY OF THYMOSIN BETA 4 (FORMULATED AS RGN-532) BY INTRAVENOUS INJECTION IN RATS APPENDIX 10 (PAGE 5): FETAL SEX, VITAL STATUS AND BODY WEIGHT - INDIVIDUAL DATA ------------------------------------------------------------------------------------------------------------------------------------ GROUP 3 RGN-352 60 MG/KG/DAY ------------------------------------------------------------------------------------------------------------------------------------ FETUS # 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 ------------------------------------------------------------------------------------------------------------------------------------ RAT # CLs 4051 4/ 9 FA FA FA MA / FA MA MA MA FA FA FA FA FA 5.56 5.50 5.78 6.08 5.45 5.78 5.55 5.54 5.78 4.89 4.88 5.49 5.29 4052 11/ 5 MA MA E FA MA FA FA FA FA MA MA / FA MA FA FA MA 5.12 5.66 5.46 5.47 5.71 5.28 5.06 5.42 5.44 5.81 5.34 5.50 5.27 5.13 5.55 4053 4/ 9 FA FA FA FA / MA MA MA FA FA FA MA E FA 5.89 6.14 5.56 5.44 5.83 6.15 6.10 5.79 5.71 5.81 5.92 5.80 4054 6/ 6 FA MA MA FA FA MA / MA FA MA MA MA MA 5.88 6.36 5.90 5.89 5.80 5.97 6.12 5.73 6.01 6.36 6.33 6.22 4055 8/ 8 FA FA MA FA MA MA E MA / MA FA MA FA MA FA FA MA 4.65 5.88 5.08 5.22 5.74 5.92 5.86 5.47 5.23 5.62 5.39 5.56 4.78 5.19 5.95 4056 7/ 5 FA MA MA FA FA E FA / MA FA MA FA MA 5.64 5.91 6.28 6.23 5.61 5.79 5.85 5.57 5.95 5.46 5.90 4057 4/ 9 FA MA FA E / MA MA MA FA FA FA FA FA MA 5.23 6.59 6.02 6.47 6.39 6.06 6.09 5.61 5.68 5.81 5.32 6.44 4058 7/ 6 FA FA FA MA MA FA MA / MA MA FA MA FA FA 4.52 5.12 5.37 5.23 5.30 4.92 5.22 5.49 5.33 5.28 5.37 5.29 5.29 4059 4/ 8 MA E MA MA / MA FA MA FA MA MA FA MA 6.42 6.03 6.46 4.10 5.77 6.00 6.02 5.96 6.12 5.88 6.16 4060 8/ 9 MA MA MA FA MA FA FA / MA MA FA FA FA E MA MA 5.78 6.05 5.72 5.56 6.09 6.12 5.89 5.87 5.82 6.06 5.35 5.84 6.25 6.35 4061 10/ 7 E FA MA FA FA FA FA FA MA FA / FA MA FA FA MA FA 5.10 5.90 5.21 5.63 5.78 5.05 5.13 5.79 5.59 5.62 6.35 5.51 5.62 5.86 5.73 4062 11/ 5 FA MA FA FA FA FA FA MA FA MA / FA FA FA FA FA 5.23 5.57 5.84 5.47 6.02 5.31 5.81 5.59 5.41 5.75 5.59 5.58 5.62 5.88 5.79 4063 10/ 6 MA MA MA MA FA MA FA FA MA MA / MA FA MA FA MA MA 5.89 6.02 5.69 5.93 5.68 6.20 5.73 5.92 6.43 6.35 6.02 5.73 6.00 5.89 6.16 6.49 4064 8/ 8 MA MA FA FA FA MA MA MA / MA MA MA MA FA E E 5.43 5.92 5.45 5.74 5.87 5.92 5.74 6.31 5.91 6.03 5.85 5.86 5.32 4065 9/ 5 MA MA MA FA FA MA MA MA MA / MA MA MA MA MA 5.46 5.61 5.71 5.75 5.70 5.81 5.90 5.75 5.56 5.52 5.71 5.98 6.09 5.92 4066 10/ 5 FA MA FA FA MA FA MA E FA MA / FA FA MA MA 5.89 6.15 6.04 6.04 6.62 6.19 6.53 5.79 6.79 6.31 6.41 6.80 6.70 4067 8/ 8 FA FA MA FA FA FA MA FA / MA FA FA FA FA FA MA 5.29 5.37 5.62 5.31 5.62 5.78 5.81 5.88 5.99 5.98 5.86 5.29 6.11 6.24 6.02 4068 7/ 7 MA FA MA MA MA MA / MA MA MA MA MA FA FA 5.80 4.78 6.00 5.79 5.79 5.42 5.32 5.69 5.79 6.02 5.79 5.69 5.71 ------------------------------------------------------------------------------------------------------------------------------------ M = MALE F = FEMALE A = ALIVE E = EARLY RESORPTION L = LATE RESORPTION "/" DENOTES POSITION OF CERVIX CLs = CORPORA LUTEA/OVARY FETAL BODY WEIGHTS WERE RECORDED IN GRAMS (G).


Revised Draft Report Page 183 Testing Facility Study No. 20024504 PROTOCOL 20024504: AN EMBRYO-FETAL DEVELOPMENT STUDY OF THYMOSIN BETA 4 (FORMULATED AS RGN-532) BY INTRAVENOUS INJECTION IN RATS APPENDIX 10 (PAGE 6): FETAL SEX, VITAL STATUS AND BODY WEIGHT - INDIVIDUAL DATA ------------------------------------------------------------------------------------------------------------------------------------ GROUP 3 RGN-352 60 MG/KG/DAY ------------------------------------------------------------------------------------------------------------------------------------ FETUS # 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 ------------------------------------------------------------------------------------------------------------------------------------ RAT # CLs 4069 9/ 5 MA FA MA FA FA MA FA FA MA / FA MA MA MA MA 5.97 5.86 6.01 5.82 5.69 5.90 5.61 5.82 6.15 5.55 5.94 6.30 5.92 6.19 4070 9/ 7 MA MA E MA MA FA MA FA FA / FA MA FA MA MA MA MA 5.88 5.89 5.49 5.69 5.43 5.41 5.49 5.93 4.49 5.57 5.59 5.53 6.00 5.87 6.07 4071 7/ 8 FA MA FA MA FA FA FA / FA FA MA MA MA MA MA FA 5.90 6.47 5.94 6.24 6.12 5.70 6.05 6.31 5.93 6.70 6.27 6.26 6.24 6.41 6.46 4072 10/ 7 FA FA MA MA MA MA MA MA E MA / MA MA MA FA FA FA MA 4.74 5.53 5.52 5.66 5.59 5.71 5.29 5.42 5.60 5.46 5.46 5.62 5.33 5.31 5.40 5.79 4073 7/ 8 MA MA FA FA MA MA E / FA FA MA MA MA FA MA E 5.87 6.05 5.50 5.69 6.17 6.09 6.12 5.32 5.85 6.27 5.60 5.85 6.42 4074 8/ 2 MA MA FA FA MA MA MA FA / 6.08 6.30 6.19 6.07 6.44 6.54 6.30 6.34 4075 10/ 6 MA FA FA FA FA MA FA FA MA / MA FA MA FA FA MA 5.49 5.39 5.66 5.64 5.97 5.99 6.05 5.27 5.58 5.64 5.50 6.02 5.53 5.45 6.30 ------------------------------------------------------------------------------------------------------------------------------------ M = MALE F = FEMALE A = ALIVE E = EARLY RESORPTION L = LATE RESORPTION "/" DENOTES POSITION OF CERVIX CLs = CORPORA LUTEA/OVARY FETAL BODY WEIGHTS WERE RECORDED IN GRAMS (G).


Revised Draft Report Page 184 Testing Facility Study No. 20024504 PROTOCOL 20024504: AN EMBRYO-FETAL DEVELOPMENT STUDY OF THYMOSIN BETA 4 (FORMULATED AS RGN-532) BY INTRAVENOUS INJECTION IN RATS APPENDIX 10 (PAGE 7): FETAL SEX, VITAL STATUS AND BODY WEIGHT - INDIVIDUAL DATA ------------------------------------------------------------------------------------------------------------------------------------ GROUP 4 RGN-352 180 MG/KG/DAY ------------------------------------------------------------------------------------------------------------------------------------ FETUS # 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 ------------------------------------------------------------------------------------------------------------------------------------ RAT # CLs 4076 6/ 8 MA FA MA MA FA FA / MA E FA MA FA MA FA 5.96 5.30 6.31 5.65 5.75 5.64 5.19 5.34 5.58 5.40 6.06 4.86 4077 NOT PREGNANT 4078 7/ 5 FA FA FA FA MA FA FA / MA MA MA MA MA 5.95 5.87 5.61 6.05 5.96 5.73 5.82 5.95 6.02 6.28 5.92 6.02 4079 8/ 5 FA FA FA FA MA E MA FA / FA FA MA MA FA 5.39 5.22 6.06 5.87 5.53 5.90 5.91 5.82 5.75 5.41 5.99 5.82 4080 10/ 4 MA FA MA FA FA FA FA E MA MA / FA MA MA 5.02 5.19 5.12 5.53 5.25 5.81 5.45 5.69 6.04 5.67 5.52 6.06 4081 8/ 6 MA MA MA MA FA FA MA MA / MA MA MA FA MA FA 5.99 5.85 6.05 5.12 4.79 5.42 5.67 5.91 5.63 5.55 5.64 5.30 5.63 5.42 4082 6/10 MA MA MA MA MA MA / MA MA FA FA MA FA MA FA FA FA 5.82 5.66 5.77 5.78 5.41 6.00 5.95 5.83 5.27 5.25 5.61 5.37 5.91 5.15 5.98 6.08 4083 6/10 FA FA FA FA FA FA / FA MA FA FA MA FA FA MA MA FA 4.94 4.87 5.15 5.11 5.15 5.33 5.22 5.13 5.12 5.04 5.29 5.18 4.71 5.04 5.08 5.23 4084 8/ 8 FA FA FA FA MA MA MA MA / FA FA MA FA MA FA FA MA 5.00 5.18 5.01 5.25 5.54 5.28 5.47 6.04 5.26 4.93 4.97 5.19 5.28 5.31 5.03 5.73 4085 7/ 7 E MA MA FA MA FA MA / MA MA FA MA FA FA MA 6.29 6.15 6.10 6.00 6.18 6.07 6.30 6.21 5.80 6.28 5.73 6.35 6.28 4086 9/ 3 FA MA MA MA FA MA MA FA MA / FA FA 5.89 4.75 6.63 6.66 6.09 5.96 6.21 4.66 6.61 6.11 6.60 4087 7/ 7 FA MA FA FA FA MA FA / FA MA MA MA MA MA FA 5.49 5.98 5.85 5.53 5.29 6.04 5.66 5.80 6.12 6.43 6.33 6.09 6.32 5.53 4088 6/10 FA MA FA FA MA FA / FA MA FA MA FA FA FA FA MA 5.84 6.12 5.61 6.18 5.86 5.74 5.50 5.81 5.92 5.83 6.16 5.70 5.80 5.95 6.47 4089 8/ 5 FA MA MA FA MA MA FA MA / MA MA FA FA MA 5.97 6.19 6.23 5.57 6.16 6.59 5.99 6.79 6.36 6.38 5.92 6.39 6.27 4090 7/ 7 FA MA FA MA MA MA FA / MA MA FA MA FA FA MA 4.72 5.62 5.40 5.58 5.62 5.92 5.26 5.52 5.59 5.09 5.81 5.23 5.73 5.94 4091 7/ 8 MA MA FA FA MA MA / MA FA MA MA FA MA FA MA 5.95 6.23 5.80 5.66 6.10 5.89 6.17 5.85 5.88 6.31 5.80 6.14 5.90 5.72 4092 9/ 4 MA FA FA MA FA FA FA FA FA / FA FA MA MA 4.91 4.65 5.16 5.27 5.21 5.36 5.18 5.28 5.22 4.86 4.89 4.89 5.53 4093 10/ 6 MA E MA MA FA E MA FA MA MA / FA MA FA FA MA 6.70 6.15 6.54 5.61 6.42 5.46 6.47 5.64 6.28 5.38 5.93 5.90 5.77 ------------------------------------------------------------------------------------------------------------------------------------ M = MALE F = FEMALE A = ALIVE E = EARLY RESORPTION L = LATE RESORPTION "/" DENOTES POSITION OF CERVIX CLs = CORPORA LUTEA/OVARY FETAL BODY WEIGHTS WERE RECORDED IN GRAMS (G).


Revised Draft Report Page 185 Testing Facility Study No. 20024504 PROTOCOL 20024504: AN EMBRYO-FETAL DEVELOPMENT STUDY OF THYMOSIN BETA 4 (FORMULATED AS RGN-532) BY INTRAVENOUS INJECTION IN RATS APPENDIX 10 (PAGE 8): FETAL SEX, VITAL STATUS AND BODY WEIGHT - INDIVIDUAL DATA ------------------------------------------------------------------------------------------------------------------------------------ GROUP 4 RGN-352 180 MG/KG/DAY ------------------------------------------------------------------------------------------------------------------------------------ FETUS # 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 ------------------------------------------------------------------------------------------------------------------------------------ RAT # CLs 4094 8/ 9 MA MA FA FA FA MA MA MA / MA FA FA FA MA MA MA E FA 5.95 5.82 5.73 6.04 5.81 5.80 6.23 6.03 5.00 5.28 5.82 5.43 5.57 6.01 5.70 6.36 4095 7/10 FA FA MA FA FA MA FA / FA MA FA MA FA FA FA MA MA MA 5.65 5.60 6.25 5.81 5.63 6.16 5.77 5.61 5.58 5.29 5.72 5.48 5.38 5.28 5.63 5.63 6.08 4096 10/ 7 MA MA FA MA E MA FA FA MA FA / MA FA MA MA FA MA MA 5.63 6.03 5.37 5.61 4.56 5.22 3.95 5.97 5.39 6.13 5.58 5.63 5.45 5.41 5.77 5.94 4097 8/ 6 FA MA FA MA FA FA FA MA / MA MA MA FA MA MA 5.57 5.54 5.76 5.59 2.65 5.41 5.58 5.59 5.41 5.89 6.23 5.12 5.55 5.87 4098 NOT PREGNANT 4099 8/ 8 FA FA FA MA FA FA MA MA / MA MA FA MA FA FA FA FA 5.57 5.90 5.50 6.58 5.92 5.74 6.01 6.28 5.89 6.20 6.00 6.24 5.97 5.81 5.79 6.05 4100 7/12 E FA MA MA MA E E / MA MA MA FA MA MA MA FA MA FA FA 5.54 6.50 6.12 6.62 5.71 6.22 5.95 5.84 5.61 5.63 6.22 4.53 5.83 5.60 5.85 ------------------------------------------------------------------------------------------------------------------------------------ M = MALE F = FEMALE A = ALIVE E = EARLY RESORPTION L = LATE RESORPTION "/" DENOTES POSITION OF CERVIX CLs = CORPORA LUTEA/OVARY FETAL BODY WEIGHTS WERE RECORDED IN GRAMS (G).



Appendix 11 Fetal Alterations - Individual Data


Revised Draft Report Page 187 Testing Facility Study No. 20024504 PROTOCOL 20024504: AN EMBRYO-FETAL DEVELOPMENT STUDY OF THYMOSIN BETA 4 (FORMULATED AS RGN-532) BY INTRAVENOUS INJECTION IN RATS APPENDIX 11 (PAGE 1): FETAL ALTERATIONS - INDIVIDUAL DATA ------------------------------------------------------------------------------------------------------------------------------------ GROUP 1 CONTROL ARTICLE 0 (CONTROL) MG/KG/DAY ------------------------------------------------------------------------------------------------------------------------------------ LIVE SPECIMENS GROSS EXTERNAL EXAMINATION SOFT TISSUE EXAMINATION SKELETAL EXAMINATION WITH ANY RAT ALTERATIONS NUMBER N(%) N/N DESCRIPTION N/N DESCRIPTION N/N DESCRIPTION ------------------------------------------------------------------------------------------------------------------------------------ 4001 0( 0.0) 0/13 0/ 6 0/ 7 4002 1( 8.3) 0/12 0/ 6 1/ 6 FETUS 7 RIBS: INCOMPLETELY OSSIFIED, left 10th 4003 0( 0.0) 0/13 0/ 6 0/ 7 4004 0( 0.0) 0/17 0/ 8 0/ 9 4005 0( 0.0) 0/15 0/ 7 0/ 8 4006 0( 0.0) 0/15 0/ 7 0/ 8 4007 0( 0.0) 0/13 0/ 6 0/ 7 4008 1( 7.7) 0/13 0/ 6 1/ 7 FETUS 1 THORACIC VERTEBRAE: CENTRUM, BIFID, 12th 4009 1( 7.1) 0/14 0/ 7 1/ 7 FETUS 3 RIBS: WAVY, right 11th 4010 1( 7.1) 0/14 FETUS 16 1/ 7 FETUS 13 0/ 7 LATE RESORPTION, VESSELS: AORTA PASSES autolysis precluded DORSAL TO THE TRACHEA further evaluation AND ESOPHAGUS; INNOMINATE ARTERY ABSENT; ARISE IN INCORRECT ORDER, 1st left carotid, 2nd right carotid, 3rd right subclavian, 4th left subclavian ------------------------------------------------------------------------------------------------------------------------------------ N/N = NUMBER OF LIVE SPECIMENS WITH ALTERATIONS/NUMBER OF LIVE SPECIMENS EXAMINED


Revised Draft Report Page 188 Testing Facility Study No. 20024504 PROTOCOL 20024504: AN EMBRYO-FETAL DEVELOPMENT STUDY OF THYMOSIN BETA 4 (FORMULATED AS RGN-532) BY INTRAVENOUS INJECTION IN RATS APPENDIX 11 (PAGE 2): FETAL ALTERATIONS - INDIVIDUAL DATA ------------------------------------------------------------------------------------------------------------------------------------ GROUP 1 CONTROL ARTICLE 0 (CONTROL) MG/KG/DAY ------------------------------------------------------------------------------------------------------------------------------------ LIVE SPECIMENS GROSS EXTERNAL EXAMINATION SOFT TISSUE EXAMINATION SKELETAL EXAMINATION WITH ANY RAT ALTERATIONS NUMBER N(%) N/N DESCRIPTION N/N DESCRIPTION N/N DESCRIPTION ------------------------------------------------------------------------------------------------------------------------------------ 4011 0( 0.0) 0/16 0/ 8 0/ 8 4012 1( 9.1) 0/11 0/ 5 1/ 6 FETUS 8 CERVICAL VERTEBRAE: ARCH, 6TH HAS THE APPEARANCE LIKE THE 7TH, left 4013 0( 0.0) 0/14 0/ 7 0/ 7 4014 2( 18.2) 0/11 0/ 5 2/ 6 FETUS 1 RIBS: INCOMPLETELY OSSIFIED, right 10th; left 11th FETUS 11 CERVICAL VERTEBRAE: CERVICAL RIB PRESENT AT 7TH CERVICAL VERTEBRA, left 4015 0( 0.0) 0/12 0/ 6 0/ 6 4016 0( 0.0) 0/13 0/ 6 0/ 7 4017 0( 0.0) 0/14 0/ 7 0/ 7 4018 0( 0.0) 0/16 0/ 8 0/ 8 4019 0( 0.0) 0/12 0/ 6 0/ 6 4020 0( 0.0) 0/12 0/ 6 0/ 6 ------------------------------------------------------------------------------------------------------------------------------------ N/N = NUMBER OF LIVE SPECIMENS WITH ALTERATIONS/NUMBER OF LIVE SPECIMENS EXAMINED


Revised Draft Report Page 189 Testing Facility Study No. 20024504 PROTOCOL 20024504: AN EMBRYO-FETAL DEVELOPMENT STUDY OF THYMOSIN BETA 4 (FORMULATED AS RGN-532) BY INTRAVENOUS INJECTION IN RATS APPENDIX 11 (PAGE 3): FETAL ALTERATIONS - INDIVIDUAL DATA ------------------------------------------------------------------------------------------------------------------------------------ GROUP 1 CONTROL ARTICLE 0 (CONTROL) MG/KG/DAY ------------------------------------------------------------------------------------------------------------------------------------ LIVE SPECIMENS GROSS EXTERNAL EXAMINATION SOFT TISSUE EXAMINATION SKELETAL EXAMINATION WITH ANY RAT ALTERATIONS NUMBER N(%) N/N DESCRIPTION N/N DESCRIPTION N/N DESCRIPTION ------------------------------------------------------------------------------------------------------------------------------------ 4021 0( 0.0) 0/14 0/ 8a 0/ 7a 4022 0( 0.0) 0/15 0/ 7 0/ 8 4023 0( 0.0) 0/12 0/ 6 0/ 6 4024 1( 9.1) 0/11 0/ 5 1/ 6 FETUS 10 CERVICAL VERTEBRAE: ARCH, 6TH HAS THE APPEARANCE LIKE THE 7TH, left 4025 0( 0.0) 0/11 0/ 5 0/ 6 ------------------------------------------------------------------------------------------------------------------------------------ N/N = NUMBER OF LIVE SPECIMENS WITH ALTERATIONS/NUMBER OF LIVE SPECIMENS EXAMINED a. Fetus 4021-13 was examined for soft tissue alterations of the head only after fixation; a soft tissue examination of the body was not performed. Fetus 4021-14 was examined for soft tissue alterations of the body, was then fixed in alcohol and examined for skeletal alterations.


Revised Draft Report Page 190 Testing Facility Study No. 20024504 PROTOCOL 20024504: AN EMBRYO-FETAL DEVELOPMENT STUDY OF THYMOSIN BETA 4 (FORMULATED AS RGN-532) BY INTRAVENOUS INJECTION IN RATS APPENDIX 11 (PAGE 4): FETAL ALTERATIONS - INDIVIDUAL DATA ------------------------------------------------------------------------------------------------------------------------------------ GROUP 2 RGN-352 20 MG/KG/DAY ------------------------------------------------------------------------------------------------------------------------------------ LIVE SPECIMENS GROSS EXTERNAL EXAMINATION SOFT TISSUE EXAMINATION SKELETAL EXAMINATION WITH ANY RAT ALTERATIONS NUMBER N(%) N/N DESCRIPTION N/N DESCRIPTION N/N DESCRIPTION ------------------------------------------------------------------------------------------------------------------------------------ 4026 0( 0.0) 0/13 0/ 6 0/ 7 4027 0( 0.0) 0/14 0/ 7 0/ 7 4028 0( 0.0) 0/ 7 0/ 3 0/ 4 4029 0( 0.0) 0/13 0/ 6 0/ 7 4030 0( 0.0) 0/13 0/ 6 0/ 7 4031 0( 0.0) 0/14 0/ 7 0/ 7 4032 0( 0.0) 0/15 0/ 7 0/ 8 4033 1( 6.2) 0/16 0/ 8 1/ 8 FETUS 9 THORACIC VERTEBRAE: CENTRUM, BIFID, 12th 4034 0( 0.0) 0/16 0/ 8 0/ 8 4035 0( 0.0) 0/14 0/ 7 0/ 7 4036 1( 5.9) 0/17 0/ 8 1/ 9 FETUS 13 RIBS: INCOMPLETELY OSSIFIED, right 11th 4037 2( 14.3) 0/14 0/ 7 2/ 7 FETUS 1 RIBS: WAVY, right 10th and 11th; INCOMPLETELY OSSIFIED, right 10th FETUS 3 RIBS: WAVY, right 5th - 8th, 10th - 12th; left 5th - 7th, 10th - 12th; INCOMPLETELY OSSIFIED, right 11th; left 10th ------------------------------------------------------------------------------------------------------------------------------------ N/N = NUMBER OF LIVE SPECIMENS WITH ALTERATIONS/NUMBER OF LIVE SPECIMENS EXAMINED


Revised Draft Report Page 191 Testing Facility Study No. 20024504 PROTOCOL 20024504: AN EMBRYO-FETAL DEVELOPMENT STUDY OF THYMOSIN BETA 4 (FORMULATED AS RGN-532) BY INTRAVENOUS INJECTION IN RATS APPENDIX 11 (PAGE 5): FETAL ALTERATIONS - INDIVIDUAL DATA ------------------------------------------------------------------------------------------------------------------------------------ GROUP 2 RGN-352 20 MG/KG/DAY ------------------------------------------------------------------------------------------------------------------------------------ LIVE SPECIMENS GROSS EXTERNAL EXAMINATION SOFT TISSUE EXAMINATION SKELETAL EXAMINATION WITH ANY RAT ALTERATIONS NUMBER N(%) N/N DESCRIPTION N/N DESCRIPTION N/N DESCRIPTION ------------------------------------------------------------------------------------------------------------------------------------ 4038 1( 10.0) 0/10 0/ 5 1/ 5 FETUS 1 THORACIC VERTEBRAE: CENTRUM, BIFID, 12th 4039 0( 0.0) 0/12 0/ 6 0/ 6 4040 0( 0.0) 0/12 0/ 6 0/ 6 4041 2( 12.5) 0/16 1/ 8 FETUS 6 1/ 8 FETUS 15 VESSELS: INNOMINATE THORACIC VERTEBRAE: ARTERY ABSENT CENTRUM, BIFID, 13th 4042 0( 0.0) 0/13 0/ 6 0/ 7 4043 2( 18.2) 0/11 0/ 5 2/ 6 FETUS 3 STERNAL CENTRA: DUPLICATED a, 1st - 3rd, XIPHOID: DUPLICATED a, FETUS 7 CERVICAL VERTEBRAE: CERVICAL RIB PRESENT AT 7TH CERVICAL VERTEBRA, right 4044 0( 0.0) 0/17 0/ 8 0/ 9 ------------------------------------------------------------------------------------------------------------------------------------ N/N = NUMBER OF LIVE SPECIMENS WITH ALTERATIONS/NUMBER OF LIVE SPECIMENS EXAMINED a. Excluded from ossification site summarization and statistical analyses.


Revised Draft Report Page 192 Testing Facility Study No. 20024504 PROTOCOL 20024504: AN EMBRYO-FETAL DEVELOPMENT STUDY OF THYMOSIN BETA 4 (FORMULATED AS RGN-532) BY INTRAVENOUS INJECTION IN RATS APPENDIX 11 (PAGE 6): FETAL ALTERATIONS - INDIVIDUAL DATA ------------------------------------------------------------------------------------------------------------------------------------ GROUP 2 RGN-352 20 MG/KG/DAY ------------------------------------------------------------------------------------------------------------------------------------ LIVE SPECIMENS GROSS EXTERNAL EXAMINATION SOFT TISSUE EXAMINATION SKELETAL EXAMINATION WITH ANY RAT ALTERATIONS NUMBER N(%) N/N DESCRIPTION N/N DESCRIPTION N/N DESCRIPTION ------------------------------------------------------------------------------------------------------------------------------------ 4045 2( 18.2) 0/11 0/ 5 2/ 6 FETUS 1 SKULL: EYE SOCKET, SMALL, left; CERVICAL VERTEBRAE: ARCH, INCOMPLETELY OSSIFIED, left 4th; ARCHES, FUSED, right 4th and 5th; THORACIC VERTEBRAE: CENTRUM, BIFID, 11th and 12th; CENTRUM, UNILATERAL OSSIFICATION, left 5th; RIBS: FUSED, right 4th and 5th, bases, proximal - distal; left 11th and, 12th, medially; THORACIC VERTEBRAE: ARCH, SMALL, right 5th; STERNAL CENTRA: ASYMMETRIC, 1st and 2nd FETUS 7 CERVICAL VERTEBRAE: ARCH, INCOMPLETELY OSSIFIED, right 4th and 5th 4046 1( 10.0) 0/10 0/ 5 1/ 5 FETUS 9 STERNAL CENTRA: INCOMPLETELY OSSIFIED, 1st 4047 0( 0.0) 0/15 0/ 7 0/ 8 ------------------------------------------------------------------------------------------------------------------------------------ N/N = NUMBER OF LIVE SPECIMENS WITH ALTERATIONS/NUMBER OF LIVE SPECIMENS EXAMINED


Revised Draft Report Page 193 Testing Facility Study No. 20024504 PROTOCOL 20024504: AN EMBRYO-FETAL DEVELOPMENT STUDY OF THYMOSIN BETA 4 (FORMULATED AS RGN-532) BY INTRAVENOUS INJECTION IN RATS APPENDIX 11 (PAGE 7): FETAL ALTERATIONS - INDIVIDUAL DATA ------------------------------------------------------------------------------------------------------------------------------------ GROUP 2 RGN-352 20 MG/KG/DAY ------------------------------------------------------------------------------------------------------------------------------------ LIVE SPECIMENS GROSS EXTERNAL EXAMINATION SOFT TISSUE EXAMINATION SKELETAL EXAMINATION WITH ANY RAT ALTERATIONS NUMBER N(%) N/N DESCRIPTION N/N DESCRIPTION N/N DESCRIPTION ------------------------------------------------------------------------------------------------------------------------------------ 4048 2( 12.5) 0/16 0/ 8 2/ 8 FETUS 7 RIBS: INCOMPLETELY OSSIFIED, right 11th FETUS 15 RIBS: WAVY, right 7th, 8th and 11th; INCOMPLETELY OSSIFIED, right 10th 4049 1( 6.2) 0/16 0/ 8 1/ 8 FETUS 7 RIBS: WAVY, right 9th - 11th; INCOMPLETELY OSSIFIED, right 11th 4050 0( 0.0) 0/15 0/ 7 0/ 8 ------------------------------------------------------------------------------------------------------------------------------------ N/N = NUMBER OF LIVE SPECIMENS WITH ALTERATIONS/NUMBER OF LIVE SPECIMENS EXAMINED


Revised Draft Report Page 194 Testing Facility Study No. 20024504 PROTOCOL 20024504: AN EMBRYO-FETAL DEVELOPMENT STUDY OF THYMOSIN BETA 4 (FORMULATED AS RGN-532) BY INTRAVENOUS INJECTION IN RATS APPENDIX 11 (PAGE 8): FETAL ALTERATIONS - INDIVIDUAL DATA ------------------------------------------------------------------------------------------------------------------------------------ GROUP 3 RGN-352 60 MG/KG/DAY ------------------------------------------------------------------------------------------------------------------------------------ LIVE SPECIMENS GROSS EXTERNAL EXAMINATION SOFT TISSUE EXAMINATION SKELETAL EXAMINATION WITH ANY RAT ALTERATIONS NUMBER N(%) N/N DESCRIPTION N/N DESCRIPTION N/N DESCRIPTION ------------------------------------------------------------------------------------------------------------------------------------ 4051 0( 0.0) 0/13 0/ 6 0/ 7 4052 0( 0.0) 0/15 0/ 7 0/ 8 4053 0( 0.0) 0/12 0/ 6 0/ 6 4054 0( 0.0) 0/12 0/ 6 0/ 6 4055 1( 6.7) 0/15 0/ 7 1/ 8 FETUS 3 PELVIS: PUBIS, INCOMPLETELY OSSIFIED, bilateral 4056 0( 0.0) 0/11 0/ 5 0/ 6 4057 0( 0.0) 0/12 0/ 6 0/ 6 4058 0( 0.0) 0/13 0/ 6 0/ 7 4059 0( 0.0) 0/11 0/ 5 0/ 6 4060 1( 7.1) 0/14 0/ 7 1/ 7 FETUS 7 CERVICAL VERTEBRAE: CERVICAL RIB PRESENT AT 7TH CERVICAL VERTEBRA, right ------------------------------------------------------------------------------------------------------------------------------------ N/N = NUMBER OF LIVE SPECIMENS WITH ALTERATIONS/NUMBER OF LIVE SPECIMENS EXAMINED


Revised Draft Report Page 195 Testing Facility Study No. 20024504 PROTOCOL 20024504: AN EMBRYO-FETAL DEVELOPMENT STUDY OF THYMOSIN BETA 4 (FORMULATED AS RGN-532) BY INTRAVENOUS INJECTION IN RATS APPENDIX 11 (PAGE 9): FETAL ALTERATIONS - INDIVIDUAL DATA ------------------------------------------------------------------------------------------------------------------------------------ GROUP 3 RGN-352 60 MG/KG/DAY ------------------------------------------------------------------------------------------------------------------------------------ LIVE SPECIMENS GROSS EXTERNAL EXAMINATION SOFT TISSUE EXAMINATION SKELETAL EXAMINATION WITH ANY RAT ALTERATIONS NUMBER N(%) N/N DESCRIPTION N/N DESCRIPTION N/N DESCRIPTION ------------------------------------------------------------------------------------------------------------------------------------ 4061 0( 0.0) 0/15 0/ 7 0/ 8 4062 0( 0.0) 0/15 0/ 7 0/ 8 4063 0( 0.0) 0/16 0/ 8 0/ 8 4064 0( 0.0) 0/13 0/ 6 0/ 7 4065 1( 7.1) 0/14 0/ 7 1/ 7 FETUS 9 THORACIC VERTEBRAE: CENTRUM, BIFID, 11th 4066 1( 7.7) 0/13 0/ 6 1/ 7 FETUS 5 RIBS: WAVY, right 4th - 11th; left 10th; INCOMPLETELY OSSIFIED, right 5th, 8th - 12th; left 10th - 12th 4067 0( 0.0) 0/15 0/ 7 0/ 8 4068 0( 0.0) 0/13 0/ 6 0/ 7 4069 0( 0.0) 0/14 0/ 7 0/ 7 4070 0( 0.0) 0/15 0/ 7 0/ 8 4071 0( 0.0) 0/15 0/ 7 0/ 8 4072 0( 0.0) 0/16 0/ 8 0/ 8 4073 0( 0.0) 0/13 0/ 6 0/ 7 4074 0( 0.0) 0/ 8 0/ 4 0/ 4 4075 0( 0.0) 0/15 0/ 7 0/ 8 ------------------------------------------------------------------------------------------------------------------------------------ N/N = NUMBER OF LIVE SPECIMENS WITH ALTERATIONS/NUMBER OF LIVE SPECIMENS EXAMINED


Revised Draft Report Page 196 Testing Facility Study No. 20024504 PROTOCOL 20024504: AN EMBRYO-FETAL DEVELOPMENT STUDY OF THYMOSIN BETA 4 (FORMULATED AS RGN-532) BY INTRAVENOUS INJECTION IN RATS APPENDIX 11 (PAGE 10): FETAL ALTERATIONS - INDIVIDUAL DATA ------------------------------------------------------------------------------------------------------------------------------------ GROUP 4 RGN-352 180 MG/KG/DAY ------------------------------------------------------------------------------------------------------------------------------------ LIVE SPECIMENS GROSS EXTERNAL EXAMINATION SOFT TISSUE EXAMINATION SKELETAL EXAMINATION WITH ANY RAT ALTERATIONS NUMBER N(%) N/N DESCRIPTION N/N DESCRIPTION N/N DESCRIPTION ------------------------------------------------------------------------------------------------------------------------------------ 4076 0( 0.0) 0/12 0/ 6 0/ 6 4077 NOT PREGNANT 4078 0( 0.0) 0/12 0/ 6 0/ 6 4079 0( 0.0) 0/12 0/ 6 0/ 6 4080 0( 0.0) 0/12 0/ 6 0/ 6 4081 0( 0.0) 0/14 0/ 7 0/ 7 4082 0( 0.0) 0/16 0/ 8 0/ 8 4083 0( 0.0) 0/16 0/ 8 0/ 8 4084 0( 0.0) 0/16 0/ 8 0/ 8 4085 0( 0.0) 0/13 0/ 6 0/ 7 4086 0( 0.0) 0/11 0/ 5 0/ 6 ------------------------------------------------------------------------------------------------------------------------------------ N/N = NUMBER OF LIVE SPECIMENS WITH ALTERATIONS/NUMBER OF LIVE SPECIMENS EXAMINED


Revised Draft Report Page 197 Testing Facility Study No. 20024504 PROTOCOL 20024504: AN EMBRYO-FETAL DEVELOPMENT STUDY OF THYMOSIN BETA 4 (FORMULATED AS RGN-532) BY INTRAVENOUS INJECTION IN RATS APPENDIX 11 (PAGE 11): FETAL ALTERATIONS - INDIVIDUAL DATA ------------------------------------------------------------------------------------------------------------------------------------ GROUP 4 RGN-352 180 MG/KG/DAY ------------------------------------------------------------------------------------------------------------------------------------ LIVE SPECIMENS GROSS EXTERNAL EXAMINATION SOFT TISSUE EXAMINATION SKELETAL EXAMINATION WITH ANY RAT ALTERATIONS NUMBER N(%) N/N DESCRIPTION N/N DESCRIPTION N/N DESCRIPTION ------------------------------------------------------------------------------------------------------------------------------------ 4087 4( 28.6) 0/14 0/ 7 4/ 7 FETUS 1 RIBS: SHORT, left 13th FETUS 3 RIBS: WAVY, right 4th - 12th; left 5th, 9th - 13th; INCOMPLETELY OSSIFIED, right 9th - 11th; left 9th - 12th FETUS 9 RIBS: WAVY, right 10th; left 6th, 8th - 10th; INCOMPLETELY OSSIFIED, right 10th; left 8th - 11th FETUS 11 SKULL: NASAL - FRONTAL, SUTURE LARGE, bilateral; RIBS: WAVY, right 10th and 11th; INCOMPLETELY OSSIFIED, right 10th; left 11th; THICKENED, right 9th, medially 4088 0( 0.0) 0/15 0/ 7 0/ 8 4089 0( 0.0) 0/13 0/ 6 0/ 7 4090 0( 0.0) 0/14 0/ 7 0/ 7 4091 0( 0.0) 0/14 0/ 7 0/ 7 ------------------------------------------------------------------------------------------------------------------------------------ N/N = NUMBER OF LIVE SPECIMENS WITH ALTERATIONS/NUMBER OF LIVE SPECIMENS EXAMINED


Revised Draft Report Page 198 Testing Facility Study No. 20024504 PROTOCOL 20024504: AN EMBRYO-FETAL DEVELOPMENT STUDY OF THYMOSIN BETA 4 (FORMULATED AS RGN-532) BY INTRAVENOUS INJECTION IN RATS APPENDIX 11 (PAGE 12): FETAL ALTERATIONS - INDIVIDUAL DATA ------------------------------------------------------------------------------------------------------------------------------------ GROUP 4 RGN-352 180 MG/KG/DAY ------------------------------------------------------------------------------------------------------------------------------------ LIVE SPECIMENS GROSS EXTERNAL EXAMINATION SOFT TISSUE EXAMINATION SKELETAL EXAMINATION WITH ANY RAT ALTERATIONS NUMBER N(%) N/N DESCRIPTION N/N DESCRIPTION N/N DESCRIPTION ------------------------------------------------------------------------------------------------------------------------------------ 4092 0( 0.0) 0/13 0/ 7a 0/ 6 4093 0( 0.0) 0/13 0/ 6 0/ 7 4094 0( 0.0) 0/16 0/ 8 0/ 8 4095 1( 5.9) 0/17 0/ 8 1/ 9 FETUS 15 CERVICAL VERTEBRAE: CERVICAL RIB PRESENT AT 7TH CERVICAL VERTEBRA, left 4096 0( 0.0) 0/16 0/ 8 0/ 8 4097 1( 7.1) 0/14 0/ 7 1/ 7 FETUS 9 RIBS: INCOMPLETELY OSSIFIED, right 11th 4098 NOT PREGNANT 4099 0( 0.0) 0/16 0/ 8 0/ 8 4100 0( 0.0) 0/15 0/ 7 0/ 8 ------------------------------------------------------------------------------------------------------------------------------------ N/N = NUMBER OF LIVE SPECIMENS WITH ALTERATIONS/NUMBER OF LIVE SPECIMENS EXAMINED a. The body of fetus 4092-9 was examined for soft tissue alterations after fixation.



Appendix 12 Historical Control Data


SUMMARY OF REPRODUCTIVE INDICESCrl:CD(SD) RATS

DAY 21 CAESAREAN-SECTION

PERIOD: JANUARY 2010 - JANUARY 2012

FULL DOSAGE-RANGE STUDIES STUDIES

NUMBER OF STUDIES 36 12

NUMBER OF RATS:TESTED 898 96PREGNANT 850 94FOUND DEAD 3 0ABORTED 1 0DELIVERED 6 0

NUMBER OF RATS PREGNANT AT CAESAREAN-SECTIONING 842 94

NUMBER OF RATS WITH SINGLE CONCEPTUS LITTER:LIVE 1 1RESORBED 0 0ABORTED 0 0

RANGE/STUDY RANGE/STUDY MEAN or % MEAN or % MEAN or % MEAN or %

% PREGNANT 95.0 (80.0-100) 98.1 (87.5-100) AVERAGE NO. CORPORA LUTEA 15.4 (14.1-17.3) 15.2 (13.4-16.8) AVERAGE NO. IMPLANTATIONS 14.6 (13.4-16.4) 14.2 (11.7-15.6) AVERAGE % PREIMPLANTATION LOSS 5.1 (1.1-10.0) 5.9 (0.7-18.5)

AVERAGE LITTER SIZE

AVERAGE NO. LIVE FETUSES 13.9 (12.4-16.0) 13.5 (11.4-15.1) AVERAGE NO. DEAD FETUSES 0.0 (0-0.2) 0.0 (0-0.2)

AVERAGE NO. RESORPTIONS 0.7 (0.2-1.1) 0.7 (0.3-1.0) AVERAGE NO. EARLY RESORPTIONS 0.6 (0.2-1.1) 0.6 (0.3-1.0) AVERAGE NO. LATE RESORPTIONS 0.0 (0-0.4) 0.0 (0-0.1)

AVERAGE % POSTIMPLANTATION LOSS 4.8 (1.7-12.2) 5.8 (1.9-18.7)


SUMMARY OF REPRODUCTIVE INDICESCrl:CD(SD) RATS



RANGE/STUDY RANGE/STUDY MEAN or % MEAN or % MEAN or % MEAN or %

AVERAGE % DAMS WITH ANY RESORPTIONS 42.4 (21.7-68.2) 44.7 (25.0-75.0)

AVERAGE % DAMS WITH ALL CONCEPTUSES RESORBED 0.1 (0-4.5) 1.1 (0-12.5)

AVERAGE % DAMS WITH ONE OR MORE LIVE FETUSES 99.9 (95.4-100) 98.9 (87.5-100)

AVERAGE SEX RATIO (% MALES/LITTER) 49.4 (42.6-55.1) 50.4 (39.3-61.0) AVERAGE FETAL BODY WEIGHT (G) 5.54 (5.30-5.75) 5.55 (5.31-5.87) AVERAGE FOR MALES (G) 5.69 (5.44-5.92) 5.70 (5.45-6.03)

AVERAGE FOR FEMALES (G) 5.39 (5.13-5.61) 5.39 (5.11-5.76)

AVERAGE % DEAD OR RESORBED CONCEPTUSES/LITTER 4.7 (1.7-8.2) 4.8 (1.9-8.9)


SUMMARY OF MATERNAL NECROPSY OBSERVATIONS Crl:CD(SD) RATS - DAY 21 CAESAREAN-SECTION


NO. STUDIES 45NO. RATS TESTED 919NO. RATS PREGNANT 870NO. RATS DIED 6NO. RATS ABORTED 1NO. RATS DELIVERED 6NO. RATS WITH 100% RESORPTION 1

OBSERVATIONS MEAN RANGE/ STUDYN % N %

TRACHEADorsal, proximal to heart, dark red gelatinous material present (found dead) 1 0.11 0-1 (0-4.0)

LIVERMedian lobe, constricted area 1 0.11 0-1 (0-4.0)

THORACIC CAVITYRed and/or cloudy fluid present (euthanized or found dead) 1 0.11 0-1 (0-4.0)

SPLEENAdhered to abdominal adipose and stomach 1 0.11 0-1 (0-4.0)Discolored 1 0.11 0-1 (0-4.0)Large 2 0.22 0-1 (0-4.0)

KIDNEY(S)Pelvis, slight dilation 1 0.11 0-1 (0-4.0)Pelvis, moderate dilation 1 0.11 0-1 (0-4.2)

STOMACHTissue present (delivered early) 2 0.22 0-1 (0-4.0)

INTESTINESJejunum; diverticulum 1 0.11 0-1 (0-12.5)

URETERSSlight dilation, bilateral 1 0.11 0-1 (0-4.2)


SUMMARY OF MATERNAL NECROPSY OBSERVATIONS Crl:CD(SD) RATS - DAY 21 CAESAREAN-SECTION

OBSERVATIONS MEAN RANGE/ STUDYN % N %

UTERUSHorns filled with clear, brown fluid 1 0.11 0-1 (0-4.0)Uterus elongated, proximal to cervix; uterine adipose tissue, red; left horn, mucosal and serosal surface, dark red and contained dark red fluid and red gelatinous material 1 0.11 0-1 (0-12.5)Bilateral, slight hydrometra 1 0.11 0-1 (0-4.0)

OVARIESLeft, dark red 1 0.11 0-1 (0-12.5)Right, red bursal cyst 1 0.11 0-1 (0-4.0)


LITTER AND FETAL ALTERATIONS SUMMARY Crl:CD(SD) RATS DAY 21 CAESAREAN-SECTION


NUMBER OF STUDIES: 30

NUMBER OF LITTERS EVALUATED: 663

NUMBER OF FETUSES EVALUATED (LIVE): 9092

RANGE/STUDY MEAN or % MEAN or %

LITTERS WITH FETUSES WITHANY ALTERATION OBSERVED % 22.6 (0-50.0)

FETUSES WITH ANYALTERATION OBSERVED % 2.2 (0-4.3)

% FETUSES WITH ANYALTERATION/LITTER MEAN 2.2 (0-4.6)


SUMMARY OF FETAL GROSS EXTERNAL ALTERATIONS Crl:CD(SD) RATS - DAY 21 CAESAREAN-SECTION



NO. STUDIES INCLUDED 36 12NO. LITTERS EXAMINED 840 94NO. LIVE FETUSES EXAMINED 11673 1272

ALTERATION RANGE /STUDY RANGE /STUDYN % N % N % N %

HEADExencephaly L 1 0.12 0-1 (0-4.3) 0 --- --- ---

F 1 0.01 0-1 (0-0.3) 0 --- --- ---

EYE One or both eye bulges L 3 0.36 0-1 (0-5.0) 0 --- --- --- depressed F 3 0.03 0-1 (0-0.4) 0 --- --- ---

SNOUT Cleft L 1 0.12 0-1 (0-4.3) 0 --- --- ---

F 1 0.01 0-1 (0-0.3) 0 --- --- ---

NARESAbsent L 1 0.12 0-1 (0-5.3) 0 --- --- ---

F 1 0.01 0-1 (0-0.4) 0 --- --- ---

PALATE Cleft L 2 0.24 0-1 (0-5.0) 0 --- --- ---

F 2 0.02 0-1 (0-0.4) 0 --- --- ---

TONGUEProtruded L 1 0.12 0-1 (0-4.3) 0 --- --- ---

F 1 0.01 0-1 (0-0.3) 0 --- --- ---

JAW Micrognathia L 1 0.12 0-1 (0-4.0) 0 --- --- ---

F 1 0.01 0-1 (0-0.3) 0 --- --- ---

BODY Umbilical hernia L 3 0.36 0-1 (0-4.3) 0 --- --- ---

F 3 0.03 0-1 (0-0.3) 0 --- --- ---

L: LITTER INCIDENCEF: FETAL INCIDENCE


SUMMARY OF FETAL GROSS EXTERNAL ALTERATIONS Crl:CD(SD) RATS - DAY 21 CAESAREAN-SECTION


ALTERATION RANGE /STUDY RANGE /STUDYN % N % N % N %

BODY (CONT.)Anus, no opening L 1 0.12 0-1 (0-4.0) 0 --- --- --- present F 1 0.01 0-1 (0-0.2) 0 --- --- ---Edema L 3 0.36 0-1 (0-4.3) 0 --- --- ---

F 3 0.03 0-1 (0-0.3) 0 --- --- ---Spina bifida L 1 0.12 0-1 (0-4.3) 0 --- --- ---

F 1 0.01 0-1 (0-0.3) 0 --- --- ---Trunk short L 1 0.12 0-1 (0-5.3) 0 --- --- ---

F 1 0.01 0-1 (0-0.4) 0 --- --- ---Gastroschisis L 1 0.12 0-1 (0-4.0) 0 --- --- ---

F 1 0.01 0-1 (0-0.3) 0 --- --- ---

FORE AND/OR HINDLIMB(S)Digit(s), short L 1 0.12 0-1 (0-5.3) 0 --- --- ---

F 1 0.01 0-1 (0-0.4) 0 --- --- ---Digit(s), absent L 2 0.24 0-1 (0-4.2) 0 --- --- ---

F 2 0.02 0-1 (0-0.3) 0 --- --- ---Digit(s), extra L 1 0.12 0-1 (0-4.0) 0 --- --- ---

F 1 0.01 0-1 (0-0.3) 0 --- --- ---Limb(s), short L 2 0.24 0-1 (0-5.3) 0 --- --- ---

F 2 0.02 0-1 (0-0.4) 0 --- --- ---Limb(s), rotated L 3 0.36 0-1 (0-5.3) 0 --- --- ---

F 3 0.03 0-1 (0-0.4) 0 --- --- ---Limb(s), extra L 1 0.12 0-1 (0-4.0) 0 --- --- ---

F 1 0.01 0-1 (0-0.3) 0 --- --- ---

TAILThreadlike L 2 0.24 0-2 (0-10.0) 0 --- --- ---

F 2 0.02 0-2 (0-0.6) 0 --- --- ---Absent L 1 0.12 0-1 (0-4.0) 0 --- --- ---

F 1 0.01 0-1 (0-0.2) 0 --- --- ---Curved L 1 0.12 0-1 (0-4.2) 0 --- --- ---

F 1 0.01 0-1 (0-0.3) 0 --- --- ---Short L 1 0.12 0-1 (0-5.3) 0 --- --- ---

F 1 0.01 0-1 (0-0.4) 0 --- --- ---



SUMMARY OF FETAL SOFT TISSUE ALTERATIONS Crl:CD(SD) RATS



NO. STUDIES INCLUDED 30NO. LITTERS EXAMINED 662NO. FETUSES EXAMINED 4456

RANGE/STUDY ALTERATION N % N %

BRAINLateral ventricles, slight L 1 0.15 0-1 (0-4.2) dilation F 1 0.02 0-1 (0-0.6)

EYERetina folded in one or both L 5 0.76 0-2 (0-9.1)

F 5 0.11 0-2 (0-1.4)

PALATEInvaginated L 1 0.15 0-1 (0-4.0)

F 1 0.02 0-1 (0-0.6)

TONGUESmall L 1 0.15 0-1 (0-4.0)

F 1 0.02 0-1 (0-0.6)

HEARTInterventricular septal defect L 1 0.15 0-1 (0-4.2)

F 1 0.02 0-1 (0-0.6)

VESSELSInnominate artery, absent L 1 0.15 0-1 (0-4.0)

F 1 0.02 0-1 (0-0.7)

LUNGOne or more lobes, partial or L 2 0.30 0-1 (0-4.2) complete agenesis F 2 0.04 0-1 (0-0.6)Right diaphragmatic and L 1 0.15 0-1 (0-4.2) apical lobes fused F 1 0.02 0-1 (0-0.6)

THORACIC AND/OR ABDOMINAL CAVITIESSitus inversus L 1 0.15 0-1 (0-4.0)

F 1 0.02 0-1 (0-0.6)



SUMMARY OF FETAL SOFT TISSUE ALTERATIONS Crl:CD(SD) RATS


RANGE/STUDY ALTERATION N % N %

URINARY BLADDERCyst L 1 0.15 0-1 (0-4.2)

F 1 0.02 0-1 (0-0.6)



SUMMARY OF FETAL SKELETAL ALTERATIONS Crl:CD(SD) RATS - DAY 21 CAESAREAN-SECTION

PERIOD: JANUARY 2010 - JANUARY 2012NO. STUDIES INCLUDED 29NO. LITTERS EXAMINED 655NO. FETUSES EXAMINED 4635

RANGE/STUDYALTERATION N % N %

SKULLFrontals: suture, large L 1 0.15 0-1 (0-4.2)

F 1 0.02 0-1 (0-0.6)Nasal - Frontal: suture, large L 9 1.37 0-2 (0-8.3)

F 14 0.30 0-3 (0-1.9)Parietals: incompletely ossified L 2 0.30 0-1 (0-4.2)

F 2 0.04 0-1 (0-0.6)Eye socket: small L 3 0.46 0-1 (0-4.3)

F 3 0.06 0-1 (0-0.6)Palate: incompletely ossified L 2 0.30 0-1 (0-4.3)

F 2 0.04 0-1 (0-0.6)Premaxilla: incompletely ossified L 1 0.15 0-1 (0-4.0)

F 1 0.02 0-1 (0-0.6) : short L 1 0.15 0-1 (0-4.2)

F 1 0.02 0-1 (0-0.6)Maxilla: incompletely ossified L 1 0.15 0-1 (0-4.3)

F 1 0.02 0-1 (0-0.6) : short L 1 0.15 0-1 (0-4.2)

F 1 0.02 0-1 (0-0.6)Squamosal: incompletely ossified L 4 0.61 0-1 (0-4.2)

F 4 0.09 0-1 (0-0.6)Zygomatic: incompletely ossified L 4 0.61 0-1 (0-4.2)

F 4 0.09 0-1 (0-0.6)

VERTEBRAECervical: Arch, incompletely L 15 2.29 0-2 (0-25.0) ossified F 16 0.34 0-2 (0-3.3) : Arch, 6th had appearance L 15 2.29 0-3 (0-12.5) like 7th F 17 0.37 0-3 (0-1.9) : Arch, 7th had appearance L 6 0.92 0-2 (0-8.3) like 6th F 6 0.13 0-2 (0-1.1) : Arches, fused L 2 0.30 0-1 (0-4.8)

F 2 0.04 0-1 (0-0.7) : Hemivertebra L 2 0.30 0-1 (0-4.3)

F 2 0.04 0-1 (0-0.6)





VERTEBRAE (CONT.)Thoracic: Centrum, bifid L 52 7.94 0-5 (0-25.0)

F 55 1.19 0-5 (0-3.3) : Centrum, unilateral L 4 0.61 0-1 (0-4.3) ossification F 4 0.09 0-1 (0-0.6) : Centrum, incompletely or L 2 0.30 0-1 (0-4.3) not ossified F 2 0.04 0-1 (0-0.6) : Arches, fused L 1 0.15 0-1 (0-4.0)

F 1 0.02 0-1 (0-0.6) : Arch, small L 2 0.30 0-1 (0-4.3)

F 2 0.04 0-1 (0-0.6) : Hemivertebra L 1 0.15 0-1 (0-4.3)

F 1 0.02 0-1 (0-0.6)

Lumbar: Centrum, bifid L 1 0.15 0-1 (0-4.2)F 1 0.02 0-1 (0-0.6)

: Centrum, incompletely or L 1 0.15 0-1 (0-4.3) not ossified F 1 0.02 0-1 (0-0.6) : Arch, incompletely ossified L 2 0.30 0-1 (0-4.2)

F 2 0.04 0-1 (0-0.6) : 7 present L 1 0.15 0-1 (0-4.0)

F 3 0.06 0-3 (0-1.7)

Sacral: Centrum, incompletely L 1 0.15 0-1 (0-4.3) ossified F 1 0.02 0-1 (0-0.6)

Caudal: Centrum, incompletely L 1 0.15 0-1 (0-4.3) ossified F 1 0.02 0-1 (0-0.6)

RIBS Cervical rib(s) present L 38 5.80 0-6 (0-24.0)

F 46 0.99 0-10 (0-5.5)One or more, wavy L 12 1.83 0-1 (0-4.5)

F 13 0.28 0-2 (0-1.3)One or more incompletely ossified L 7 1.07 0-2 (0-8.3) (hypoplastic) or not ossified F 10 0.22 0-3 (0-1.8)Fused L 3 0.46 0-1 (0-4.2)

F 3 0.06 0-1 (0-0.6)Short L 18 2.75 0-3 (0-12.5)

F 18 0.39 0-3 (0-1.8)





RIBS (CONT.)Irregularly shaped L 1 0.15 0-1 (0-4.0)

F 1 0.02 0-1 (0-0.6)Thickened L 11 1.68 0-2 (0-8.3)

F 13 0.28 0-3 (0-1.7)Split L 1 0.15 0-1 (0-4.3)

F 1 0.02 0-1 (0-0.6)12 present L 1 0.15 0-1 (0-4.3)

F 1 0.02 0-1 (0-0.6)Two segments L 1 0.15 0-1 (0-4.2)

F 1 0.02 0-1 (0-0.6)

STERNEBRAEOne or more incompletely L 14 2.14 0-2 (0-12.5) ossified or not ossified F 16 0.34 0-2 (0-1.7)One or more asymmetric L 6 0.92 0-1 (0-4.3)

F 6 0.13 0-1 (0-0.6)Irregularly shaped L 3 0.46 0-1 (0-4.3)

F 3 0.06 0-1 (0-0.6)Fused L 3 0.46 0-1 (0-4.2)

F 3 0.06 0-1 (0-0.6)Close-set L 1 0.15 0-1 (0-4.0)

F 1 0.02 0-1 (0-0.6)Duplicated L 3 0.46 0-1 (0-4.2)

F 3 0.06 0-1 (0-0.6)Large L 1 0.15 0-1 (0-4.2)

F 1 0.02 0-1 (0-0.6)

MANUBRIUMIrregularly shaped L 3 0.46 0-1 (0-4.2)

F 3 0.06 0-1 (0-0.6)Fused L 1 0.15 0-1 (0-4.0)

F 1 0.02 0-1 (0-0.6)Large L 2 0.30 0-1 (0-4.3)

F 2 0.04 0-1 (0-0.6)Duplicated L 2 0.30 0-1 (0-4.2)

F 2 0.04 0-1 (0-0.6)

XIPHOIDDuplicated L 2 0.30 0-1 (0-4.0)

F 2 0.04 0-1 (0-0.6)Irregularly shaped L 1 0.15 0-1 (0-4.0)

F 1 0.02 0-1 (0-0.5)





SCAPULAEBody and Ala, bent L 1 0.15 0-1 (0-4.3)

F 1 0.02 0-1 (0-0.6)

PELVISPubis, incompletely ossified L 8 1.22 0-2 (0-8.3)

F 10 0.22 0-2 (0-1.3)Ischium, incompletely ossified L 5 0.76 0-2 (0-8.3)

F 7 0.15 0-3 (0-1.6)

FORELIMB(S)Humerus, short L 1 0.15 0-1 (0-4.3)

F 1 0.02 0-1 (0-0.6)Radius and ulna, bent L 1 0.15 0-1 (0-4.3)

F 1 0.02 0-1 (0-0.6) : separated L 1 0.15 0-1 (0-4.0)

F 1 0.02 0-1 (0-0.6) : small L 1 0.15 0-1 (0-4.0)

F 1 0.02 0-1 (0-0.6)Phalanx, absent L 1 0.15 0-1 (0-4.0)

F 1 0.02 0-1 (0-0.6)Digit, absent L 1 0.15 0-1 (0-4.0)

F 1 0.02 0-1 (0-0.6)Metacarpal, absent L 1 0.15 0-1 (0-4.0)

F 1 0.02 0-1 (0-0.6)

HINDLIMB(S)Femur, tibia and fibula, bent L 1 0.15 0-1 (0-4.3)

F 1 0.02 0-1 (0-0.6)Digit: extra L 1 0.15 0-1 (0-4.0)

F 1 0.02 0-1 (0-0.6) : absent L 1 0.15 0-1 (0-4.2)

F 1 0.02 0-1 (0-0.6)Phalanx: extra L 1 0.15 0-1 (0-4.0)

F 1 0.02 0-1 (0-0.6) : not ossified L 1 0.15 0-1 (0-4.2)

F 1 0.02 0-1 (0-0.6)Metatarsal, extra L 1 0.15 0-1 (0-4.0)

F 1 0.02 0-1 (0-0.6)



SUMMARY OF FETAL OSSIFICATION SITE AVERAGES SKELETAL AVERAGES Crl:CD(SD) RATS DAY 21 CAESAREAN-SECTION


NO. STUDIES INCLUDED 29NO. LITTERS EXAMINED 654NO. FETUSES EXAMINED 4628

FETUS/LITTERSKELETAL AVERAGES MEAN RANGE/STUDY

HYOID 0.99 (0.93-1.00)VERTEBRAE CERVICAL 7.00 -- THORACIC 13.09 (13.00-13.18) LUMBAR 5.90 (5.80-6.00) SACRAL 3.00 -- CAUDAL 7.53 (7.01-8.48)RIBS (pairs) 13.07 (13.00-13.12)STERNUM MANUBRIUM 1.00 -- STERNAL CENTERS 3.99 (3.97-4.02) XIPHOID 1.00 (0.98-1.00)FOREPAWS (Calculated as average per limb) CARPALS 0.00 -- METACARPALS 4.00 (3.98-4.00) DIGITS 5.00 -- PHALANGES 8.13 (7.57-8.78)HINDPAWS (Calculated as average per limb) TARSALS 0.03 (0-0.09) METATARSALS 4.90 (4.81-4.98) DIGITS 5.00 -- PHALANGES 6.45 (5.62-7.68)



Appendix 13 Tabulated Summary for The Common Technical

Document (CTD)


The Common Technical Document — Safety____________________________________________________________________________________________

2.6.7.13 Reproductive and Developmental Toxicity - Effects on Embryo-fetal Development

Report Title: An Embryo-fetal Development Study of Thymosin beta 4 (formulated as RGN-352) Administered by Intravenous Injection in Rats

Test Article: Thymosin beta 4 (Tβ4) formulated as RGN-352 (Tβ4 Injectable Solution)

Design similar to ICH 4.1.3? Yes Species/Strain: Rat/Sprague-Dawley Initial Age: 66 days

Duration of Dosing: DG 7 through 17 Study No. 20024504 Location in CTD: Vol.! Page !

Date of First Dose: 15 May 2012 Day of Mating: DG 0

Special Features: None Day of C-Section: DG 21 No Observed Adverse Effect Level:

F0 Females: 180 mg/kg/day F1 Litters: 180 mg/kg/day

Method of Administration: Intravenous injection Vehicle/Formulation: Aqueous solution

GLP Compliance: Yes

Daily Dose (mg/kg) 0 (Control) 20 60 180

Dams: No. Pregnant 25 25 25 23 No. Died or Sacrificed Moribund 0 0 0 0 No. Aborted or with Total Resorption of Litter 0 0 0 0 Clinical Observations - - - - Necropsy Observations - - - - Body Weight DG 18 (%)a 365.8 g -0.8 0.0 +1.9 Food Consumption DG 7-18 (%)a 24.7 g/day +2.0 +0.8 +1.2 Mean No. Corpora Lutea 14.7 15.2 14.6 14.9 Mean No. Implantations 14.4 14.3 14.1 14.6 Mean % Preimplantation Loss 1.5 4.9 3.0 2.0 - No noteworthy findings DG = Day of presumed gestation a. At end of dosing period. For controls, group means are shown. For treated groups, percent differences from controls are shown. Statistical significance is

based on actual data (not on the percent differences).


The Common Technical Document —Safety____________________________________________________________________________________________

2.6.7.13 Reproductive and Developmental Toxicity - Effects on Embryo-fetal Development

(Continued) Study No. 20024504

Daily Dose (mg/kg) 0 (Control) 20 60 180

Litters: No. Litters Evaluated 25 25 25 23 No. Live Fetuses 13.3 13.6 13.5 14.1 Mean No. Resorptions 1.1 0.7 0.6 0.5 No. of Litters with Dead Fetuses 0 0 0 0 Mean % Postimplantation Loss 7.3 5.2 4.1 3.1 Mean Fetal Body Weight (g) 5.54 5.59 5.79** 5.71 Fetal Sex Ratios (% males) 46.1 46.0 52.7 51.4 Fetal Anomalies:

Gross External - - - - Visceral Anomalies - - - - Skeletal Anomalies - - - -

Total Affected Fetuses (Litters) 8 (7) 15## (10) 4 (4) 6 (3) - No noteworthy findings DG = Day of presumed gestation Dunnett’s Test ** - p<0.01 Variance Test for Homogeneity of the Binomial Distribution ## - p<0.01


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