Introduction to Enteris BioPharma

September 2014

Enteris BioPharma

• Privately held, New Jersey based biotech company

• Owned by Victory Park Capital, a Chicago based investment firm

• Clinically validated oral formulation technology

• for peptides and challenging small molecules

• Extensive scientific know-how and R&D experience

• Proven GMP tablet manufacturing capabilities

2

Enteris BioPharma

• Enteris has effectively addressed both permeability and solubility challenges with a simple, elegant and scalable solution

• Demonstrated a track record of clinical success across a range of compounds and therapeutic indications

• Enteris offers robust IP protection, regulatory CMC support and finished, solid dosage formulations for preclinical and early phase clinical studies

3

Clinically Validated Oral Delivery Technology

• Clinically validated oral peptide delivery technology • Positive Phase 3 oral Calcitonin: Osteoporosis(1)

• Positive Phase 2 oral PTH: Osteoporosis(2)

• Positive Phase 2 oral Calcitonin: Osteopenia(3) • Positive Phase 1 oral CR845: Neuropathic Pain(4)

• Sponsored preclinical peptide programs

• 18 ongoing or completed formulation programs

• (1) Tarsa Therapeutics, Inc. (JBMR 27, No.8, 2012, 1821-1829) • (2) Unigene Laboratories, Inc. (Bone 53, 2013, 160-166) (Clin Pharm 52, No. 6, 2013) • (3) Tarsa Therapeutics, Inc. (ASBMR, 2012) • (4) Cara Therapeutics, Inc. (data on file)

4

0

5

10

15

20

25

0 1000 2000 3000 4000 5000 6000

Actual Bioavailability Data Peptides and Small Molecules

Molecular Weight (Da)

Abs

olut

e B

ioav

aila

bilit

y (%

)

insulin calcitonin

proprietary peptide

proprietary peptides

CR-845

proprietary peptide

zanamivir

octreotide

Studies in beagle dogs

tobramycin

5

tigecycline

PTH 1-34

proprietary peptide

kanamycin

Criteria for Selection of Peptides

6

1. Characteristics of the API

a. What is the peptide sequence (or number of amino acids)?

Are there any chemical modifications to the peptide (and if so, what are they)?

i. What is the total molecular weight (including any modifications)?

ii. Is the peptide soluble in pure water? Buffers or salt solutions (which ones)? Acidic pH?

iii. Does it aggregate?

iv. Is it susceptible to proteolysis (qualitatively)?

v. Is it cyclic? How large is the macrocycle?

vi. What is the overall net charge? The pI?

vi. Any special phys-chem properties that should be known?

2. Project status

a. Is this in clinical development?

b. What is the indication?

c. Assuming we’re successful, do you have a target date for getting a tablet formulation into the clinic?

3. Feasibility for oral delivery

a. What is the injectable dose (IV/IM/SC)?

b. What’s the mechanism of action (e.g., agonist or antagonist)?

c. What’s the therapeutic window?

Contents

• Introduction

• Mechanism

• Peptides

• Small molecules

• BCS Class III

• BCS Class II

• Safety

• Excipient Safety Profile

• LLC Toxicology Studies

• LLC Regulatory and Clinical

• Patents

• Business Development

• Manufacturing

7

Mechanism of Drug Delivery

8

Enteric Coat Prevents Tablet from Opening in Stomach at Low pH

• Acid-stable enteric coating prevents tablet release in stomach • Less susceptible to food effects

or dilution with liquids • API protected from degradation

by acid and pepsin • Peptides • Acid-labile small molecules

9

Enteric Coat Dissolves at Neutral pH in the Small Intestine

• Water-soluble sub-coat acts as a

partition layer between the enteric

coat and the acidic tablet core

• Simultaneous release of API

and excipients

10

pH Modifier, Permeability Enhancers and API Released

• Citric acid sequestered in coated beads • Increases stability of tablet formulation • Compatible with peptides and small

molecules • Acts as protease inhibitor for peptides • Calcium chelator

and membrane permeation enhancer • pH-lowering agent that increases absorptive

flux • Membrane wetting/charge dispersal agent

11

API Absorbed Across Intestinal Wall via Paracellular Transport

• Lauroyl-L-carnitine (12-carbon fatty acid) • Modulates tight junctions in the intestinal

enterocytes and enhances paracellular transport

• Acts as a solubilizing agent due to surfactant properties

• Inhibits P-gp efflux transporters

12

Components of Enteris’ Solid Dosage Formulation

13

Peptide Experience

14

Dog Model Predicts Bioavailability in Humans

0 1 2 3 4 5 0

500

1000

1500

2000

2500

3000

3500

4000

4500

5000 Linear Regression for Cmax: Y = A + B * X R 2 =0.9801

Plas

ma

Cm

ax (p

g/m

l)

Dose (mg) 10 100 1000 10000 10

100

1000 y=.995x - 0.7548 R=0.958 R 2 =0.918

Hum

an C

max

(pg/

ml)

Dog Cmax (pg/ml)

Enteris' dog model for oral delivery shows high degree of linearity with respect to dose offering a wide range of dosing strategies. Comparability of PK results in dog and human shows that Enteris’ dog model is an appropriate success predictor for human studies

15

Bioavailability of 9 Amino Acid Peptide Across Multiple Animal Models

Enteris Enteris Enteris

All D Amino Acid Peptide

16

Absorption of LHRH Analog in Dogs as a Function of Enteric-Coating

Formulation B

Unformulated C

Formulation D

Formulation Enteric coat (weight gain)

Tmax (min)

Bioavailability

(% F)

A L30D-55

(10%) 111 3.0

B L30D-55

(15%) 116 4.6

D L30D-55/

FS30D (12%) 152 7.2

Unformulated L30D-55

(10%) 130 0.1

0

2000

4000

6000

8000

10000

12000

0 100 200 300 400 500LH

RH

(pg

/mL)

Time Relative to Tmax (minutes)

Capsule Formulation in Dogs

Formulation A

Formulation B

Formulation D

Unformulated

17

Bioavailability of Cara’s CR845 in Preclinical & Phase I

0%

5%

10%

15%

20%13% 13%

16%

Rat Dog Man

18

Phase I Oral CR845 Study

Time (hours)

CR84

5 (ng

/mL)

0 4 8 12 16 20 240.1

1

10

100

3 mg

0.5 mg1 mg

10 mg

N = 8/group

Mean + SEM

CR845 Demonstrated 16% Oral Bioavailability

19

PTH Phase II Study

Mean PTH Cmax Values for Subjects Receiving Oral PTH(1-31)NH2 and Forsteo®

20

0.00

0.50

1.00

1.50

2.00

2.50

3.00

rsCT Tablet Nasal Spray Placebo

p<0.001* p=0.014* p=ns*

1.5

0.8 0.5

Phase III Oral Calcitonin Study

Mea

n %

Cha

nge

LS-B

MD

Phase III Study for Oral sCT:

Primary Endpoint (Change in LS BMD) Achieved

21

Small Molecule Experience

22

Rationale for for BCS Class II, III and IV Small Molecule Drugs

• Lauroyl-L-carnitine (12-carbon fatty acid) • Modulates tight junctions in the intestinal enterocytes and enhances

paracellular transport • Acts as a solubilizing agent due to surfactant properties • Inhibits efflux transporters (P-gp)

• Citric Acid (Organic acid) • Calcium chelator and membrane permeation enhancer • pH-lowering agent that increases absorptive flux • Membrane wetting/charge dispersal agent

23

KANAMYCIN PROJECT REVIEW AMY STURMER 1.0 / 31JULY2013

Tigecycline Case Study

24

• Currently approved only for IV infusion as a last resort antibiotic therapy • 100mg loading dose, followed by 50mg every 12hrs, duration ranging

from 5 to 14 days • Must be dosed in the clinic

• BCS Class III • Very high solubility in water: >295 mg/mL at all pH ranging from 1 – 14 • Very poor permeability: Oral formulations explored to date exhibit a limit

of approx. 5 %F • Technology is uniquely suited to enable oral formulation with suitable F

• Oral therapy offers out of clinic dosing • Reduced overall healthcare costs • Strategy: Initiate dosing by IV titration, then discharge with oral therapy

BCS Class III Molecule Tigecycline Case Study

25

BCS Class III Molecule Tigecycline Rat Study

Animals dosed via intraduodenal administration to simulate oral dosing

(0.64 mg/kg IV or 4.8 mg/kg ID)

0.0 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.00.00

0.25

0.50

0.75

1.00

1.25

1.50

1.75

2.00

2.25

IV

ID 100mM CA, 26mM LLCID PBS

ID 400mM CA, 26mM LLC

Time (hrs)

Plas

ma T

igec

yclin

e (m

cg/m

L)Pl

asm

a C

once

ntra

tion

(mcg

/mL)

IV ID PBS ID 100mM CA, 26mM LLC ID 400mM CA, 26mM LLC Cmax (mcg/mL) 1.79 0.10 0.71 1.11

Tmax (hr) 0.08 1.50 0.33 0.28

AUC(0-t) (mcg*hr/mL) 58.20 6.96 49.38 117.63

%F(0-t) -- 1.60 10.85 27.90 26

Summary of Rat Tigecycline Study

• %F for unformulated ID tigecycline: 1.60 to 2.76%

• %F of formulated ID tigecycline increased by 10 to 20 fold depending on formulation • 10.85% at 100mM CA / 26mM LLC • 27.90% at 400mM CA / 26mM LLC

• Achieved high oral bioavailability for tigecycline where other formulation technologies have failed, or only been marginally effective

27

Tigecycline Solid Dosage Form Development Enteric Coated Capsule

• BCS class III molecule filled in capsules • “Formulated”: API, CA, LLC and filler • “Unformulated”: API and filler only • Single ascending dose study in beagle dogs • 4 single dose arms

• 15 mg formulated + unformulated (n=8 dogs each) • 30 mg formulated (n=5 dogs) + unformulated (n=3 dogs) • 45 mg formulated (n=5 dogs) • 5 mg IV bolus (n=3 dogs)

28

BCS Class III Molecule (Tigecycline) Beagle Dog PK Study

• API was not detected in any “unformulated” arm

0.0 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.00

30

60

90

120

150

180

15 mg30 mg45 mg

Time (hr)

Pla

sma

Tige

cycl

ine

Con

cent

rati

on(n

g/m

L)

Plas

ma

Con

cent

ratio

n (n

g/m

L)

29

• Summary • %F of formulated ID dosing in rat model increased by 10 to 20 fold

• Enabled the development of a solid oral dosage form with desirable level of bioavailability and commercial viability in dog model

• Achieved high oral bioavailability where other formulation technologies have failed

BCS Class III Molecule (Tigecycline) Study Summary

PK Data: Enteric Coated Capsules in Dogs

Mean 5 mg IV

Mean 15 mg PO

Mean 30 mg PO

Mean 45 mg PO

Cmax (ng/mL) 335.0 75.51 121.13 177.01

Tmax (h) 0.08 1.84 1.58 1.67

AUC(0-t) (ng*hr/mL) 410.86 132.65 366.65 574.32

%Fest -- 12.2 14.87 15.53

F%CV -- 72.0 43.3 67.7

30

Synergy of CA and LLC Demonstrated on Tigecycline

Mean %F

400mM CA/26mM LLC 21.64

0 CA/26mM LLC 8.86

(m)

31

Tigecycline Human PK Data

32

KANAMYCIN PROJECT REVIEW AMY STURMER 1.0 / 31JULY2013

Aminoglycoside (Kanamycin, Tobramycin) Case Studies

33

Project Overview

• Background • Aminoglycoside bactericidal antibiotics • Consist of two amino sugars glycosidically linked to deoxystreptamine • Available in oral, IV, IM, and inhaled

• Indication: • Used to treat wide variety of infections • Effective against gram negative bacteria

• Mechanism of action: • Interacts with the 30S subunit of prokaryotic ribosomes. Induces substantial

amounts of mistranslation and indirectly inhibits translocation during protein synthesis

• Contraindications: • Side effects include tinnitus, toxicity to kidneys, and allergic reactions to the drug • Presence of intestinal obstruction • Generally not indicated for long-term therapy due to nephrotoxocity and ototoxicity

• Marketed by BMS, Alcon, Novartis etc. • Wide veterinary use for certain indications

34

Mean PK for Kanamycin Following Oral Administration to Beagle Dogs (±SEM)

Formulation Key Excipients N Cmax (ng/mL)

Tmax (min)

AUC(0-t) (ng*min/mL) %F

1 0 mg CA, 0 mg LLC, uncoated

6 67 (9) 118 (18) 9167 (983) 2.8 (0.3)

2 500 mg CA, 100 mg LLC,

coated 4 428 (88) 101 (13) 46216 (6593) 14.2 (2.0)

3 250 mg CA, 100 mg LLC,

coated 3 408 (72) 125 (22) 36970 (10465) 11.4 (3.2)

4 100 mg CA, 100 mg LLC,

coated 3 489 (196) 160 (10) 40336 (18038) 12.4 (5.5)

5 50 mg CA,

100 mg LLC, coated

31 147 (147) 195 (NA) 8573 (8573) 2.6 (2.6)

35

IV Tobramycin Study

36

Plasma Levels in 8 Dogs Given Formulated Oral Tobramycin Capsules

37

Mean Plasma Levels in Dogs Given Oral Tobramycin

(Adjusted for Tmax and dose)

38

PK of Tobramycin

Unformulated Capsules

(SEM)

Formulated Capsules

(SEM)No of Dogs/No of

Responders 3/8 8/8

Cmax (ng/mL)3

(1)314 (46)

Tmax (min)145 (35)

144 (20)

AUC(0-t) (ng*min/mL)

270 (140)

25829 (4738)

AUC(0-t) (ng*min/mL/mg)

27 (14)

2583 (474)

%F0.15

(0.08)14.5 (2.7)

39

Pharmacokinetic Parameters for Kanamycin and Tobramycin

Formulation Key Excipients N Cmax (ng/mL)

Tmax (min)

AUC(0-t) (ng*min/mL) %F

JSV-003-005 10 mg Kanamycin,

500 mg CA, 100 mg LLC

4 428 (88) 101 (13) 46216 (6593) 14.2 (2.0)

JSV-003-051 10 mg Tobramycin,

500 mg CA, 100 mg LLC

8 314 (46) 144 (20) 25829 (4738) 14.5 (2.7)

Mean Pharmacokinetic Parameters for Kanamycin and Tobramycin Following Oral Administration to Beagle Dogs (±SEM)

40

BCS Class III Molecule Zanamivir Case Study

41

BCS Class III Molecule Zanamivir Case Study

• PK studies of zanamivir in beagle dogs included the following arms: • I.V. (n=3 dogs)

• 0.083 mg/kg • dosed as 1 mL of 1 mg/mL zanamivir in PBS

• Formulated enteric-coated capsule (n=5 dogs) • 1.25 mg/kg • 15 mg zanamivir, 500 mg citric acid, 100 mg LLC, Prosolv

• Un-formulated enteric-coated capsule (n=3 dogs) • 1.25 mg/kg • 15 mg zanamivir, Prosolv

• Plasma samples collected to 4 hours

42

Zanamivir Dog Study Results

0

100

200

300

400

500

600

700

0 30 60 90 120 150 180 210 240

Zana

miv

ir pl

asm

a co

ncen

trat

ion

(ng/

mL)

Time post-dose (minutes)

IV [0.083 mg/kg]

unformulated [1.25 mg/kg]

formulated: 500 mg CA, 100 mg LLC [1.25 mg/kg]

F0-4h = 19.8% (56% CV)

43

Conclusions from Zanamivir PK Study

• Bioavailability • Unformulated: 0.95% (88% CV) • Formulated: 19.8% (56% CV) • %F is somewhat underestimated, as the elimination phase of the

oral dose is not complete at 4 hours

• Compared to Relenza® Diskhaler (marketed inhalation product, GSK)

• Relenza %F∞ ranges from 4% to 17% post-inhalation • Dosage form requires inhaler device and specially-packaged blisters

of Relenza® • High variability due to differences in inhalation performance

• Enteris technology shows ~20% bioavailability over just 4 hours

44

Small Molecule Experience BCS Class II

45

In-vitro Fenofibrate Solubility Study

• BCS class II compound • Insoluble in water • Slightly soluble in ethanol (1 mg/mL) • Soluble in DMF (30 mg/mL) and DMSO (15 mg/mL) • Solubility in 1:3 DMF:PBS pH 7.2 reported at 250 mcg/mL

• Equilibrium solubility in water and increasing levels of LLC • Excess fenofibrate weighed into individual PP vials • Solutions mixed at 125rpm at 25°C for 4 days • Fenofibrate concentration measured by HPLC against a standard curve

prepared in neat CH3CN • This experiment measured water solubility at 4.3 ng/mL

46

0.0 2.5 5.0 7.5 10.00

50

100

150

200

250

300

350

400

Solubility in 1:3 DMF:PBS pH 7.20.25mg/mL

O

ClO

CH3

CH3

O

OCH3

CH3

LLC Concentration (% w/v)

Feno

fibra

te C

once

ntra

tion

(mcg

/mL)

Fenofibrate Equilibrium Solubility with Increasing Concentrations of LLC

47

Safety

48

Excipient Safety Profile

• Protease Inhibitor: Organic Acid • Pharmaceutically accepted ingredient

• Transport Enhancer: Acyl Carnitine • Scope of non-clinical safety package has been confirmed by FDA • Genotoxicity and respiratory toxicity completed • 6 month rat toxicology study completed • 9 month dog toxicology study completed • Part of tablet formulation in 12 clinical studies (328 subjects )

• All other Excipients • Pharmaceutically accepted ingredients

• Drug Master File (Type V Safety Data) • Available for cross-reference

49

Acylcarnitines as Permeation Enhancers

• Acylcarnitines are fatty acid esters of L-carnitine that have a single aliphatic hydrocarbon chain of variable length.

• 3-O-lauroyl-L-carnitine, LLC, is the carnitine ester of lauric acid, a 12 carbon aliphatic fatty acid.

• Plasma acylcarnitine concentrations in healthy human subjects ranges from 6 μM to 15 μM, with the majority of studies reporting values ≥10 μM. • Function to transfer long chain fatty acids across the

mitochondrial membrane for β-oxidation and subsequent adenosine triphosphate (ATP) production

50

The Permeation Enhancement by LLC is Transient

LeCluyse E.L., et al. (1993). J. Pharm. Exp. Therap. 265(2):955-962. 51

LLC’s Effects are Reversible Within 15 to 30 Minutes of Removal

LeCluyse E.L., et al. (1993). J. Pharm. Exp. Therap. 265(2):955-962.

Control Tissue 2.0 mM LLC

52

Little Potential for Opportunistic “Piggyback” Permeation

Molecule Type MW (kDa) 3-Dimensional Radius (Å)

Macromolecules 1 – 10 10 – 30

LPS > 100 100 – 1000

Enteric Toxins 70 – 900

Viruses 600 – 1000

Bacteria > 1000

Modeling of perturbed membrane in Caco-2 cells indicates an effective pore radius ca. 20Å

53

Tight Junction Modifiers are Common

• Drug Compounds: • Aspirin • NSAIDS • Phenothiazines

• Food and Drug Additives/Excipients: • EDTA • C8-C18 fatty acids • Various polymers • Poly-L-lysine

• Natural/Food Products: • ZOT • ATP • Chitosan and chitosan

derivatives • Wheat gluten • Oat saponins • Capsaicin • Alcohol

54

LLC Toxicology Studies

55

3-O-Lauroyl-L-Carnitine Preclinical Safety Studies

• Completed Safety Pharmacology Studies • Acute neurotoxicity in rat • Acute respiratory in rat • Acute cardiovascular in dog • hERG • CYP450 inhibition/ induction • Metabolic profiling in hepatocytes (multiple species)

• Completed Toxicology Studies • Oral MTD studies in rat and dog • 4 day oral repeat dose finding in rat and dog • 1 month oral toxicology with toxicokinetics in rat and dog • Standard genotoxicity • 6 month oral repeat dose in rat • 9 month oral repeat dose in dog

56

All Safety Pharmacology Studies Generated Desirable Outcomes

• Lauroyl-L-Carnitine did not: • Inhibit hERG tail current in vitro, at doses up to ca. 90 μg/mL • Affect ECG parameters in dogs dosed up to 100 mg/kg/day PO

for 1 month • Produce cardiovascular effects in dogs at up to 100 mg/kg • Induce any respiratory effects in rats at up to 100 mg/kg by

gavage • Produce any adverse effects on neurobehavioral function in rats

at up to 100 mg/kg

57

LLC Regulatory and Clinical

58

FDA Feedback on LLC program

• FDA Advice Letter received August, 2009 ...the studies that have been completed to date in your development program and the proposed six- and nine-month repeat dose studies in rats and dogs respectively, would serve to support the use of LLC as an excipient in drug products.

59

• Reproductive toxicology • Studies required for populations of premenopausal females, women

of child bearing potential, or males • Embryo-fetal studies – for registration

• Recommended embryo-fetal studies in two species, with an assessment of teratogenicity as a minimum to complete drug approval applications in postmenopausal women

• Carcinogenicity – likely not required • The nonclinical findings to date could support the proposal that

carcinogenicity studies are not necessary • If there are no preneoplastic lesions or serious adverse toxicologic

effects in these studies, FDA would concur that carcinogenicity studies will not be necessary (None observed – reports submitted Sept, 2012)

Registration Requirements

60

“3-O-Lauroyl-L-Carnitine Hydrochloride (LLC) Preclinical and Clinical Data.”

• Contains full study reports of all non-clinical and

clinical studies • Non-clinical and clinical overview documents • Available to partners for cross-referencing

LLC Type V DMF Submitted to FDA

61

Summary

• Extensive preclinical toxicology package • Observed no preneoplastic lesions or serious adverse

toxicologic effects • Completed toxicology study of 9 months duration

• Extensive Clinical Experience

• < 8 single dose Phase 1 studies • 8 week Phase 2a study for oral sCT program • 24 week Phase 2 study for oral PTH program

• Type V DMF containing safety data of LLC

62

Patents

63

Patents Summary

Oral Delivery Patents

• 9 issued U.S. patents

• 2 allowed U.S. patent applications

• 3 pending U.S. provisional patent applications

• 29 issued Foreign patents

• 5 pending Foreign patent applications

• Key issued patents extend through 2030

64

Business Development

65

In-vitro testing

(days - weeks)

~$10-30K

Developability Assessment

Matrix

(hours - days)

In-vivo rat

model

(6 weeks) ~$72K

In-vivo dog

model

(6-8 weeks) ~$120-160K

Y N

Suitable candidate?

Y N

Y

N

Suitable candidate?

Suitable candidate to go direct to tablet in dog?

Enteris Feasibility Study Flowchart

Review with Client at each stage Stop any unsuitable candidates asap Focus resources on candidates with potential for success

Manufacturing

67

Manufacturing

• Enteris cGMP Manufacturing • 32,000 ft2 cGMP facility located in Boonton, NJ • Separate tableting and nasal spray filling suites • Full QA/QC and regulatory support • Commercial product in US distribution

68

Recent Technical Achievements

• Identified coated organic acid as compatible excipient with peptides and small molecules

• Simple and scaleable manufacturing process

• Optimized release characteristics and bioavailability

• Demonstrated room temperature stability of peptide tablets for 24 months

• Supplied CTM for Phase 1 and Phase 2 studies

69

Tableting and Capsuling Line

• Comil conical mill • V-Blender • Korsh XL-100 10 station tablet press - up to

10,000 tablets/hr • Natoli NP-RD10a single station press for 1 to

300 tablets • Vector LDCS coating pan for enteric coating • Capsugel Profill capsule filler • Phase 1 and 2 clinical supplies • Clinical packaging

• Open label, double blind

70

Quality Control

• Raw Material Release Testing • Final Product Release • Assay Development • Assay Transfer, Optimization & Validation • Special Projects and Investigations • Stability Studies • In process, intermediate and facility testing

capabilities

71

Quality Assurance

• Systems compliant with FDA, EMEA, MHRA and ICH • Customer focused, providing real time feedback on all quality related

issues. • Full project participation:

• process development → batch record design → GMP manufacture. • Manufacturing oversight through concurrent batch record review. • Vendor auditing and qualification program.

72

Inspection History

Regulatory Inspection History • Most recent FDA inspection – October, 2011 - No FDA-483 issued

• Successful Pre-Approval Inspection – June, 2003 • Most recent EMA inspection – October, 2006 - GMP Certificate

issued • Successful Pre-Approval Inspection – February, 1998

• GMP inspections by QPs from UK & Germany for Phase 3 - Clinical Trial – 2008 • GMP Certificates issued

73

Thank You