Introduction to Enteris BioPharma, Inc.

Enteris BioPharma

•

Privately held, New Jersey based biotech company

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Owned by Victory Park Capital, a Chicago based investment firm

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Clinically validated oral formulation technology • for peptides and challenging small molecules

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Extensive scientific know-how and R&D experience

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Proven GMP tablet manufacturing capabilities

2

Enteris BioPharma

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

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

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

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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 • 15 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

Actual Bioavailability Data Peptides and Small Molecules 25

Absolute Bioavailability (%)

proprietary peptide

20

15

zanamivir tigecycline CR-845 kanamycin tobramycin

10 proprietary peptides proprietary peptide

5

octreotide *GLP-1 analog

0

0 Studies in beagle dogs Except *Rat Study

1000

2000

calcitonin

PTH 1-34

3000

4000

insulin

5000

6000

Molecular Weight (Da) 5

Criteria for Selection of Peptides

ENTERIS PEPTIDE ASSESSMENT MATRIX Molecular Weight (Da) • Sequence of peptide • Hydrodynamic radius (Å) • Cyclic peptide?

Solubility • Acid

• Water • Buffers/salts

• FaSSIF/FeSSIF

Chemical Stability • x-S-S-x Resistant to Proteolysis?

Modified? Physical Stability • Aggregation rate Projected Dose (mg)

Injectable Dose (mg)

Net Charge • Anionic charges

Bioavailability (%) if known

• Cationic charges

• Rat

• pI

• Dog 6

Criteria for Selection of Small Molecules

ENTERIS SMALL MOLECULE ASSESSMENT MATRIX Molecular Weight (Da)

Moisture sensitive/hygroscopic

BCS Class, if known

Is the API plastic, elastic or brittle?

Describe the solid (salt form, hydrate, solvate, etc.

Any known excipient incompatibilities?

Solubility

Primary mechanism(s) of chemical degradation

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pH/Solubility curve available?

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Water

•

Buffers/Solvents/Emulsions

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If insoluble, what is the most likely reason? (hydrophobicity/crystal surface energy/etc.)

Are other salts or polymorphs available? Is the API crystalline or amorphous? •

Has polymorph transition been observed during processing or on stability

Caco-2 Permeability • Apical to Basolateral, Basolateral to Apical? Identify known transporter interactions, (P-gp, BCRP, MRP family, PepT1, OATP, etc.) What is the primary clearance mechanism(s)? (renal/Biliary/Metabolic) If metabolic, what enzymes predominate? Bioavailability of current oral formulation in development?

Please describe any PO formulation attempted thus far 7

Contents

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Introduction

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Mechanism

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Safety • Excipient Safety Profile

• Peptides

• LLC Toxicology Studies

• Small molecules

• LLC Regulatory and Clinical

• BCS Class III

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Patents

• BCS Class II

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Business Development

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Manufacturing

8

Mechanism of Drug Delivery

9

Enteric Coat Prevents Tablet from Opening in Stomach at Low pH

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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

10

Enteric Coat Dissolves at Neutral pH in the Small Intestine

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Water-soluble sub-coat acts as a partition layer between the enteric

coat and the acidic tablet core • Simultaneous release of API and excipients

11

pH Modifier, Permeability Enhancers and API Released

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Organic 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

12

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

13

Components of Enteris’ Solid Dosage Formulation

14

Peptide Experience

15

Dog Model Predicts Bioavailability in Humans 4500 4000

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

1000

Human Cmax (pg/ml)

Plasma Cmax (pg/ml)

5000

3500 3000 2500

2000 1500 1000 500 0 0

1

2

3

Dose (mg)

4

5

y=.995x - 0.7548 R=0.958 R2=0.918

100

10

10

100

1000

10000

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

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Peptide Bioavailability Formulated vs. Unformulated

Effect of Formulation on the Bioavailability of Various Peptides in Pre-Clinical Animal Models Program Type

Sponsored

No. of Amino Acids

Study Type

Ranges of Unformulated Bioavailability (%) for 12 Peptides

Ranges of Formulated Bioavailability* (%) for 12 Peptides

Rat

0.35 – 6.0

10.7 – 26.0

Dog

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

Modified from: Brayden, D. Permeation Enhancers and Oral Peptide Delivery. Presented at the Roche Colorado Peptide Symposium, Sept. 12, 2011. 55

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

56

LLC Toxicology Studies

57

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

Completed Safety Pharmacology Studies • • • • • •

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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 58

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

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LLC Regulatory and Clinical

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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 ninemonth repeat dose studies in rats and dogs respectively, would serve to support the use of LLC as an excipient in drug products.

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Registration Requirements

• 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)

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LLC Type V DMF Submitted to FDA

“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

63

Summary

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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

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Type V DMF containing safety data of LLC

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Patents

65

Patents Summary

Oral Delivery Patents •

7 issued U.S. patents

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2 allowed U.S. patent applications

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3 pending U.S. provisional patent applications

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29 issued Foreign patents

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5 pending Foreign patent applications

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Key issued patents extend through 2030

66

Business Development

67

Working with Enteris

Formulation Development

In-Vitro • Solubility • Permeability In-Vivo • Intra-duodenal: Rat Study In-Vivo • Capsule: Dog Study

Tablet Formulation Optimization

3 months

In-Vitro & In-Vivo • Stability; Design Space • Tablet: Dog Study

3 months

68

Manufacturing

69

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 70

Recent Technical Achievements

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

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Simple and scaleable manufacturing process

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Optimized release characteristics and bioavailability

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Demonstrated room temperature stability of peptide tablets for 24 months

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Supplied CTM for Phase 1 and Phase 2 studies

71

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

72

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

73

Quality Assurance

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Systems compliant with FDA, EMEA, MHRA and ICH

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Customer focused, providing real time feedback on all quality related issues.

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Full project participation: • process development → batch record design → GMP manufacture.

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Manufacturing oversight through concurrent batch record review.

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Vendor auditing and qualification program.

74

Inspection History

Regulatory Inspection History • Most recent FDA inspection – October, 2011 - No FDA-483 issued • Successful Pre-Approval Inspection – June, 2003

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Most recent EMA inspection – October, 2006 - GMP Certificate issued • Successful Pre-Approval Inspection – February, 1998

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

75

E. coli Recombinant Peptide Manufacturing

DirectExpression ExpressionofofPeptides Peptides Direct • • •

Patented E. coli-based recombinant peptide expression technology Peptide secreted directly into culture medium Enzymatic amidation of recombinant precursor

Enteris’ Peptide Manufacturing Advantages • • • •

Robust yield and purity Scalable Efficient Cost-effective

76

Enteris’ Recombinant Manufacturing Process

77

High Yield and High Product Purity at Low Cost

PTH analog-gly

Glucose regulatory peptide analog-gly

Glucose regulatory peptide-gly

kDa

sCTgly

Recombinant Peptide Manufacturing

66.2 45.0 31.0 21.5 14.4 6.5

SDS PAGE and HPLC Trace of Crude Medium • Enriched Starting Material • Extracellular yields of 400 to 1300 mg/L • Reduced purification steps 78

Amidated Peptide Manufacturing Example rhPTH(1-34)NH2 Step 1. Recombinant production of rhPTH(1-34)Gly35OH precursor The peptide: human PTH(1 -34)-Gly

rhPTH(1-34)GLY Production and Fermentation Feed Rate

SVSEIQLMHNLGKHLNSMERVENLRKKLQDVHNFG MW = 4,116.8 Da P1 P2

KAN-R

ompA

800

0.300

PTH(1-34)gly 700

Term

0.250

P1

PTH(1-34)gly

pPTH(1-34)G-03

Term

7799 bp

P3

Ori

PRLA-4

0.200 PTH(1-34) mg/L

ompA

500 400

0.150

300

Feed Rate mL/min

600

P2

0.100 200 0.050 100

Term SecE P/O LAC-IQ

SEC-E

0

0.000 1

5

9

13

17

21

25

29

Hours Post Induction

Term

The host : E. coli B strain, BLM-6

PTH(1-34) mg/L

Feed Rate ml/min

79

Thank You