Formulating Peptides/proteins for Oral Delivery: a Clinical Case

Study

John S. Vrettos, Ph.D. Sr. Principal Scientist

Formulation Development

2014 AAPS National Meeting and Exposition

Enteris BioPharma

• Clinically validated solid oral dosage formulation technology

• for peptides and challenging small molecules

• Enteris has effectively addressed both permeability and

solubility challenges with a simple, elegant and scalable solution

• Privately held, New Jersey based biotech company • Owned by Victory Park Capital, a Chicago-based investment firm • Extensive scientific know-how and R&D experience • Proven GMP tablet and API manufacturing capabilities • Demonstrated a track record of clinical success across a range of

compounds and therapeutic indications • Enteris offers robust IP protection

• Patents extend to 2030 • Feasibility study and licensing business model

2

Manufacturing

• Enteris cGMP Manufacturing

• 32,000 ft2 cGMP facility located in Boonton, NJ

• Separate tableting and nasal spray filling suites

• Multi-kilogram scale microbial fermentation and purification suites

• Full QA/QC and regulatory support

• Commercial product in US distribution

• Fortical ® Calcitonin-Salmon (rDNA origin) Nasal Spray, marketed by Upsher-Smith Laboratories

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

• Several 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

Potential Commercial Benefits of Peptelligence™

• Increases product uptake and utilization vs. parenteral administration • Increased physician starts; fewer patient refusals • Enhanced patient compliance • Better patient outcomes

• Protects and extends product exclusivity and commercial life • Provides compelling differentiation in competitive markets • Adds extra layer of robust IP protection until 2030 • Refreshes and extends commercial life of products

5

Challenges to Solid Oral Dosage Formulations

Oral peptide delivery • GI tract is designed to degrade and

digest peptides • Low permeability through the

intestinal cell layer

Peptelligence™ solution to these challenges

• Reduces peptide degradation • Increases paracellular transport • No modification of the peptide

required

Oral small molecule delivery • Solubility or dissolution limited

absorption • Poor permeability due to interaction

with efflux transporters or other mechanisms

Peptelligence™ solution to these challenges

• Permeation enhancer acts as a surfactant

• Paracellular transport bypasses transcellular permeation hurdles

6

0

5

10

15

20

25

0 1000 2000 3000 4000 5000 6000

Bioavailability Data in Dogs: Peptides and Small Molecules

Abso

lute

Bio

avai

labi

lity

(%)

Molecular Weight (Da)

SMALL MOLECULES PEPTIDES PROPRIETARY PEPTIDES

zanamivir

tobramycin

tigecycline

kanamycin

CR-845

calcitonin PTH 1-34 insulin

leuprolide

octreotide

7

Mechanism of Drug Delivery

8

Enteric Coat Dissolves at Neutral pH in the Small Intestine

• 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

• Water-soluble sub-coat acts as a

partition layer between the enteric coat and the acidic tablet core • Simultaneous release of API and

excipients

9

pH Modifier, Permeability Enhancers and API Released

• Citric acid (CA), sugar-coated granules • 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

10

API Absorbed Across Intestinal Wall via Paracellular Transport

• Lauroyl-L-carnitine (LLC) • 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 • Brittle powder, not a wax or liquid • Type V DMF on file

11

Criteria for Selection of Peptides: Developability Assessment

1. Characteristics of the API

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

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

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

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

• Does it aggregate?

• Is it susceptible to proteolysis (qualitatively)?

• Is it cyclic? How large is the macrocycle?

• What is the overall net charge? The pI?

• Any special phys-chem properties that should be known?

2. Project status

• Is this in clinical development?

• What is the indication?

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

3. Feasibility for oral delivery

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

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

• What’s the therapeutic window? 12

Pre-clinical and Clinical Peptide Experience

13

Dog Model Predicts Bioavailability in Humans

• 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

0 1 2 3 4 5 0

500

1000

1500

2000

2500

3000

3500

4000

4500

Plas

ma

Cm

ax (p

g/m

l)

Dose (mg)

Dose Linearity in Dogs (39 aa peptide)

10 100 1000 10000 10

100

1000

Hum

an C

max

(pg/

ml)

Dog Cmax (pg/ml)

Human/Dog Correlation (sCT)

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Absorption of LHRH Analog in Dogs as a Function of Enteric Coat

Formulation B

Unformulated C

Formulation D

Capsule (formulation)

Enteric coat composition (weight gain)

Tmax (min)

Bioavailability (% F)

A (+ CA/LLC)

L30D-55 (10%)

111 3.0

B (+ CA/LLC)

L30D-55 (15%)

116 4.6

C (+ CA/LLC)

L30D-55 / FS30D (12%)

152 7.2

D (– CA/LLC)

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

A

B

C

D

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Bioavailability of Cara’s CR845 in Preclinical & Phase I

0%

5%

10%

15%

20%13% 13%

16%

Rat Dog Man

16

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

17

PTH Phase II Study

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

18

Development of Oral Calcitonin for Osteoporosis

19

Pharmacokinetic and pH Profile of Enteric Coated sCT and Heidelberg Capsules in Dogs

0 30 60 90 120 150 180

0

2

4

6

8

10

Time (min)

sCT

(ng/

ml)

sCT

0

2

4

6

8Intestinal pH

pH

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Phase I sCT Exploratory Study

• Design: Single Dose, Open, Crossover Design Study • Subjects: 18 Postmenopausal Women • Study Medication Doses

• 500 μg Tablet (CA/LLC) • 1500 μg Tablet (CA) • 200 IU Fortical® Nasal Spray

• Assessments: • PD Measurements (CTX-1) up to 24 hours Post Dosing • PK Measurements up to 24 hours Post Dosing

21

sCT PK and PD Profiles Following Administration of sCT to Postmenopausal Women

0

50

100

150

200

250

0.0 1.0 2.0 3.0 4.0 5.0 6.0 7.0 8.0

sCT

(pg/

mL)

Time Relative Tmax (hours)

Nasal

500 µg PO

1500 µg PO

-100

-80

-60

-40

-20

0

20

0.0 3.0 6.0 9.0 12.0 15.0 18.0 21.0 24.0M

ean

chan

ge fr

om b

asel

ine

pl

asm

a C

TX (%

) Time (hours)

22

Dose Dependent Oral Absorption of sCT Enteric-coated Tablets in Humans

0 50 100 150 200 -20

0

20

40

60

80

100

120

140

160

180

Cm

ax P

lasm

a sC

T (p

g/m

l)

Dose sCT (µg)

Mean Cmax Individual Cmax

23

Phase II Oral sCT Dose Selection Study

• Design: Single Dose, Open, Crossover Design Study • Subjects: 24 Post Menopausal Women • Study Medication Doses:

• 50 μg Tablet (CA) • 200 μg Tablet (CA) • 200 IU Fortical® Nasal Spray

• Assessments: • PD Measurements (CTX-1) Up To 24 hours Post Dosing • PK Measurements Up To 24 hours Post Dosing

24

sCT PK and PD Profiles Following Administration of sCT to Postmenopausal Women

0

5

10

15

20

25

30

0 5 10

sCT

(pg/

mL)

Time Relative Tmax (hours)

Nasal

50 µg PO

200 µg PO

-100

-80

-60

-40

-20

0

20

0 5 10

sCT

(pg/

mL)

Time Relative Tmax (hours)

• Subsequent phase 2 study for osteopenia showed no effect from food on

PD markers (lumbar spine bone mineral density) Osteoporos Int, Volume 25, Issue 11, pages 2649-2656

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Phase 3 Trial of the Efficacy and Safety of Oral Calcitonin in Postmenopausal Osteoporosis

(ORACAL)

• Design: Double-blind, Double-dummy, Multiple Dose, Placebo-controlled, Parallel Group, 48-week

• Subjects: 565 Post Menopausal Women • Study Medication Doses:

• 200 μg Tablet (CA) • Placebo Tablet (CA) • 200 IU Fortical® Nasal Spray • Placebo Nasal Spray

• Assessments: • Primary Endpoint: Percent Change From Baseline in Bone

Mineral Density (BMD) of Axial Lumbar Spine

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Phase III Oral Calcitonin Study

Journal of Bone and Mineral Research, Volume 27, Issue 8, pages 1821-1829

Cum

ulat

ive

Per

cent

of S

ubje

cts

Percent Change in Lumbar Spine Bone Mineral Density

27

0.0

0.5

1.0

1.5

2.0

2.5

rsCT Tablet Nasal Spray Placebop=ns p=0.014 p<0.001

Phase III Oral Calcitonin Study

Mea

n %

Cha

nge

LS-B

MD

Primary Endpoint (Change in LS BMD) Achieved

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Concluding Remarks

• Enteris BioPharma has: • A clinically validated solid oral dosage formulation technology for

oral delivery of peptides

• Addressed both permeability and solubility challenges with a simple, elegant and scalable solution

• Several successful pre-clinical and clinical programs

• Cara Therapeutics CR845 (neuropathic pain)

• Tarsa Therapeutics Ostora® (osteoporosis, osteopenia)

• Proprietary and internal programs

29

Thank You!

Questions?