Development and Preclinical Evaluation of Therapeutic Radiopharmaceuticals Based on Lu-177 and Y-90...

5/26/2018 Development and Preclinical Evaluation of Therapeutic Radiopharmaceuticals Based on Lu-177 and Y-90 Labeled Monoclonal Antibodies And

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Development and preclinical evaluation of theradiopharmaceuticals based on Lu 177 and Y 90 labeantibodies

IAEAs Coordinated

Research Project (CRP)

Establishment and standardization of a technology for rea

production of cold kit formulation of DOTA-Rituximab and p

radiopharmaceuticals for labeling with Lu-177 and Y-90

Emilija Jan

Goce Delce

Repub;ic o


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PET-S+

- Auger-e-

SPECT

Camera

The Nuclear Medicine Alphabet

DIAGNOSTIC

THERAPY


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Koopmans KP, Glaudem

EJNMMMI 2012,39, S4

Tumour

1 Peptides (receptors)

2Antibodies (Antigen)

3 Other molecules (metabolic processes)

Targeted diagnosis and therapy in nuclear medicineInfection

Seite 3


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low-dose SPECT prior to therapy

Mller et al. 2013, Nucl Med Biol, in press: doi:10.1016/j.nucmedbio.2013.11.002


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ISOTOPES in Therapy =surgery with radiation

Tissuesurgery Cellsurgery Molesur

ISOTOPE

131I, 90Y, 153Sm,166Ho,177Lu

Others

E 1 3 MeV

212, 213 Bi, 211At,149Tb,

223, 224Ra

E 48 MeV

1

16

Ee fe

Range about 1 cm 30 80 m 1

-Knife -Knife AuKn


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Questions to be answered:

Realtionship between radiation dose delivered to a lesion andthe therapeutic response (in addition toxicity)

In vivo dosimetry by quantitative PET imaging

need for +-emitting metallic radionuclides

Relationship between beta energy and therapeutic responseVariation of radionuclides with different -energy

need for metallic - -emitters with very different

energy


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- emitter for therapy

Sorted along the beta energy(starting from very soft radiation 169Er) to very hard beta energy (90Most of them are produced with reactors. However, some of them neproduced with a cyclotron (67Cu).Others can be produced with a cyclotron in much better quality (186R


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PeptideAbs

Chelator

M

Receptor

Antigen

Radiometals for diagnostic imaging and therapy

Photon or positron emitters 99mTc, 68Ga, for imaging

Particle emitters 90Y, 177Lu, for radionuclide therapy

applicable only

with high specific activity and chemical purity


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90 Y

177 Lu

131I, ..153S

Nature, behavior,

RADIOPHARMACEUTICALS

THERAPY -


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linical application.177Lu-Labeled Targeted Radiotherapeutics for Treatment ofCentral Nervous System Tumors

Nature of problem and rationale for 177Lu disease sites and their constraints

A well-studied clinical application with a new radionuclide: 177Lu-MeO-DOTA-81C6

A well-studied radiotherapeutic in a new application: 177Lu-DOTA-TATE for medulloblastoma

A cocktail glass: DOTA-SIB mixing 177Lu with radiohalogens


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Two approaches for Targeted Radiotherapy of Bra

Direct Injection into SCRC Convection EnhancedDelivery

More than 200 patients with131

I-labeled 81C6 mAb

92%

6%

2%


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Potential Advantages of 177Lu for Brain Tumor Treatm

Property 131I 177Lu

Half-life (days) 8.1 6.7

Ave. -energy (keV) 182 133

Mean range (mm) 0.91 0.67

Max. range (mm) 2.3 1.8

-ray energy (keV) 364, 637 113, 208

-ray intensity (%) 81,7 7, 11

0.01

0.1

1

10

100

1000

10000

2.0 3.0 4.0 5.0

Radial distance (cm)

Absorbed

dose

(Gy)

interface

177Lu

131I

90Y

Assume 2-cm radius cavity (33.5 cm3


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Dose to Brain beyond SCRC Interface

ThresholdDose (Gy)

Brain volume (cm3) > threshold dose

131I 177Lu 90Y

110 2.05 1.02 11.70

50 4.73 1.79 18.12

10 180.3 3.64 31.14


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177Lu-DOTA-TATE vs. 90Y-DOTA-TOC

Property 90Y 177Lu

Half-life (days) 2.7 6.7

Ave. -energy (keV) 935 133

Mean range (mm) 3.9 0.67

Max. range (mm) 10.0 1.8

-ray energy (keV) none 113, 208

-ray intensity (%) none 7, 11

DOTA-TATE has 10x highaffinity vs. DOTA-TOC

Compared with 90Y, 177L

More focal field of th-particles results indamage to normal C

Imaging177

Lu -rays determination of PKpatient-specific dosi


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177Lu-DOTA-TATE: Children

Six children with relapsed or refractory neuroblastoma PET imaging with [68Ga]DOTA-TATE to determine eligibil

200 mCi 2-3 cycles i.v. at 9-wk intervals with

Co-infusion of Lys/Arg; no renal toxicity

5 of 6 patients had stabilized disease by RECIST

Gaze et al., 201


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Some results from preclinical studies..Introducing the new approach to have kiformulation for labeling

h h d f i


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The same method of preparation


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HPLC characterization before labelling

90Y-DOTATATE

In vitro stability

(EDTA, serum, etc)

Lab yield 98 %

After purification 99 %

razn molar DOTATATE: Y

Purezaradioqumica

5:1

10:1

15:1

20:1

0

50

100Sep-Pak C18

Whatman


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Animal Biodistribution in athimic mice

Sample of ROIs definition

177Lu-DOTA-hR3

20 mCi, 1,5 mg Ab

81 % RCP 95% after purif

id

d l f l i i i

d


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Two different CCK-2-receptor specific tumour-cell-lines:

AR42J exocrine rat pancreas carcinoma cells (rat CC

A431 human epidermoid carcinoma cells stably trahuman CCK-2-receptor

tota

Inte

1 nM

(BL

med

of 1

111In-DOTA-cyclo-MG1

me

BLBL

A431 AR42J

%

Aktivitt/mgPro

tein

BLBL

A431 AR42J

111In-DOTA-MG11

Peptides . development of novel targeting strategies using Lu-177 and

Cell Uptake

Cyclic minigastrin analogues - diagnosis and therapy of medullary thyroid car


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1 h p.i.

4 h p.i.

4 h p.i. + blockage(1000-fold excess ofunlabelled peptide)

111In-DOTA-cyclo-MG1

111In-DOTA-MGD5 111In-DOTA-PP-F11

111In-DOTA-MG11

Biodistribution in the A431 mouse modelTumour-Xenograft: BALB/c nude mice, A431-CCK2R human epidermoid carcinoma cells

2 x 106 cells/mouse, tumour growth ~10 days, tumour size ~0,2 g0,37 MBq 111In-DOTA-Peptide (


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Preclinical evaluation of newbiomolecules

Radiolabelled DOTA-bombesin

for diagnosis and therapy of prostate and breast cancer


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New bombesin analogue

Peptide delivered by Saudi Arabia

DOTA-Glu-Phe-Glu-Trp-Ala-Val--Ala-His-Phe-Nle-NH2

(DOTA-Glu-BNU)

Molecular weight 1670

spacer Glu-Phe-Glu (negatively charged AA)

< abdominal uptake? > renal excretion?

retained tumour uptake?

clinical potential?


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low protein binding Eppendorf tube

10-15 g DOTA-Glu-BNU (20% EtOH 1g/l)

60 l177

LuCl3 (120 MBq)+ 80 l buffer

80C,25 min

1) 1 M ascorbic acid/0.27 M gentisic acidbuffer pH 4.5

RCY =95.2%

2) 0.8 M sodium acetate buffer pH 5

RCY =99.6%

3) 0.4 M Sodium acetate/ 0.24 M gentisic acid

buffer pH 5

RCY =99.7%

4) 0.2 M 2-(N-morpholino) ethanesulfonic acid

(MES) buffer pH 5.5

RCY =5.9%

Radiolabelling with Lu-177

1)

2)

3)

0, 0 1, 3 2, 5 3, 8 5, 0 6, 3 7, 5 8, 8 10, 0 11, 3 12, 5 13, 8 15, 0 16, 3 17, 5 18, 8 20, 0 21, 3 22, 5 23, 8 25, 0 2

-50

0

100

200

300

400 Gastrin #16 [modif ied by radhplc1] 177Lu-BOMB Ask/gen mV

0, 0 1, 3 2, 5 3, 8 5, 0 6, 3 7, 5 8, 8 1 0, 0 1 1, 3 1 2, 5 13, 8 15, 0 16, 3 17, 5 18, 8 20, 0 21, 3 22, 5 23, 8 25, 0 2

-20

25

50

75

100

125

150

180 Gastrin #14 [modif ied by radhplc1] 177Lu-BOMB NaAc mV

0, 0 1, 3 2, 5 3, 8 5, 0 6, 3 7, 5 8, 8 1 0, 0 1 1, 3 1 2, 5 13, 8 15, 0 16, 3 17, 5 18, 8 20, 0 21, 3 22, 5 23, 8 25, 0 2

-50

0

50

100

150

200

250

300

350 Gastrin #15 [modif ied by radhplc1] 177Lu-BOMB NaAc/gen mV

0, 0 1, 3 2, 5 3, 8 5, 0 6, 3 7, 5 8, 8 10, 0 11, 3 12, 5 13, 8 15, 0 16, 3 17, 5 18, 8 20, 0 21, 3 22, 5 23, 8 25, 0 2

-20

0

20

40

60

80

100

120

140 Gastrin #13 [modif ied by radhplc1] 177Lu-BOMB MES mV

4)

DOTA-Glu-BNU

D bl f i N d i


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Double xenografts in Nude micespecific vs non-specific uptake


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FU-parameters, toxicitythe weight

FU-parameters, efficacy


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the tumor growthand survival time !

The double tumors imaged by


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The double tumors imaged bySPECT/CT

The double tumors imaged


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The double tumors imagedPET/MRI


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Transplanted hu tumors in

immunosuppresseed animals

Biologically similar spontaneous tum

S ll

i l

SPECT/CT i SCID

i


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Nano-SPECT/CT

of the 111In-labellel DOTA-peptidesSosabowski J (London, GB)

Small animal SPECT/CT in SCID-mice

with A431-CCK2R-tumour 4 h p.i.

Injected activity

10 MBq111

In-DOTA-Pep(0.3 nmol Pept

10-fold higher

peptide amoun

(comparable w

the therapeutic

application)

MGD5London

cyclo-MGInnsbruck

PP-F11Basel

Seite 31

Rats with SSR positive tumours in liver


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Control

Rats with SSR-positive tumours in liver

model mimics disseminated disease PRRT

177Lu-octreotate

(PRRT = Peptide Receptor Radionuclide Therapy )

Woute

ErasmusThe Netherland

S l ti it i


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-50

-40

-30

-20

-10

0

10

20

30

0 10 20 30 40

Temperature[oC]

Time [h]

Selection criteria:

- physical appearance of the obtained cake of dry product

- clear solution of reconstituted product, after reconstitution in

N t t


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Next step..

2. Freeze-dried product:

Implementation of the lyophilization protocol to thestabile product

Characterization of the lyophilized product (SEM,

DSC, ) Stability of the freeze-dried product

Ne t step


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Next step..

+ emitters for in vivo dosimetry

[111In]DTPA- Scintigraphic abi 5 &

[86Y]DOTA-DPhe1-Tyr3-

t tid


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5 h p.i.

24 h p.i.

[ ]

octreotide

SPECT

images 5 &

affe

carcin

extensive hep

paraaortal met

octreotide

PET

Patients: 3 patients with me

of carcinoid tumo

(histologically co

No therapy with u

somatostatin > 4 w

Age: 46

67 year All were candidat

a possible90Y-DOTATOC th

F.Rsch et.al.

that metallic positr

practical relevance

DOTA-SIB, A TRIFUNCTIONAL PROSTHETIC GROUP


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Long term Objective: Prosthetic moieties to label proteins and

-with two different radionuclides

-augment tumor retention after internalization

Functions:

-Chelating moiety to impart polarity/charge in addition to com

radiometals-Halobenzoyl moiety to carry radiohalogens

-Active ester for mAb/peptide conjugation

,

FOR MULTI-MODAL

LABELING


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DOTA-SIB A TRIFUNCTIONAL PROSTHETIC GROUP FOR MULTI-MOD

Ganesan Vaidyanathan, Benjamin White, Donna Affleck, Xiao-Guang Zhao, Philip Welsh, Marc Hens, and

Duke University, Durham, NC


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165Er3+

1.

2.

4.

5.

6.

7.

p+

Er

165Tm3+

165TmTmEr

165Er3+

SCX

165Tm-DOTA

165Er3+

3.

Development and Preclinical Evaluation of Therapeutic Radiopharmaceuticals Based on Lu-177 and Y-90...

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Transcript of Development and Preclinical Evaluation of Therapeutic Radiopharmaceuticals Based on Lu-177 and Y-90...