Introduction to

Enteris BioPharma, Inc.

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

• 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

0

5

10

15

20

25

0 1000 2000 3000 4000 5000 6000

Actual Bioavailability Data

Peptides and Small Molecules

Molecular Weight (Da)

Ab

so

lute

Bio

ava

ilab

ility

(%

)

insulin PTH 1-34 calcitonin

proprietary peptide

proprietary peptides

kanamycin

CR-845

proprietary peptide

zanamivir

tigecycline

octreotide

Studies in beagle dogs

Except *Rat Study 5

tobramycin

*GLP-1 analog

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

Net Charge

• Anionic charges

• Cationic charges

• pI

Chemical Stability

• x-S-S-x

Resistant to Proteolysis?

Modified?

Physical Stability

• Aggregation rate

Projected Dose (mg)

Injectable Dose (mg)

Bioavailability (%) if known

• Rat

• Dog

6

Criteria for Selection of Small Molecules

ENTERIS SMALL MOLECULE ASSESSMENT MATRIX

Molecular Weight (Da)

BCS Class, if known

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

Solubility

• pH/Solubility curve available?

• Water

• Buffers/Solvents/Emulsions

• 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

Moisture sensitive/hygroscopic

Is the API plastic, elastic or brittle?

Any known excipient incompatibilities?

Primary mechanism(s) of chemical degradation

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

• 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

8

Mechanism of Drug Delivery

9

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

10

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

11

pH Modifier, Permeability

Enhancers and API Released

• 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

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

Pla

sm

a C

ma

x (

pg/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/m

l)

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

16

Peptide Bioavailability

Formulated vs. Unformulated

Effect of Formulation on the Bioavailability of Various

Peptides in Pre-Clinical Animal Models

Program

Type

No. of

Amino

Acids

Study

Type

Ranges of Unformulated

Bioavailability (%) for 12

Peptides

Ranges of Formulated

Bioavailability* (%) for 12

Peptides

Sponsored 4 – 39

Rat 0.35 – 6.0 10.7 – 26.0

Dog <LOD** – 0.38 0.39 – 22.8

* in each case the percent bioavailability of the formulated peptide was higher than unformulated

** LOD = limit of detection

17

Bioavailability of 9 Amino Acid Peptide

Across Multiple Animal Models

Enteris Enteris Enteris

All D Amino Acid Peptide

18

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

0 100 200 300 400 500

LH

RH

(p

g/m

L)

Time Relative to Tmax (minutes)

Capsule Formulation in Dogs

Formulation A

Formulation B

Formulation D

Unformulated

19

Bioavailability of Cara’s CR845 in

Preclinical & Phase I

0%

5%

10%

15%

20%

13% 13%

16%

Rat Dog Man

20

Phase I Oral CR845 Study

Time (hours)

CR

845

(ng/

mL)

0 4 8 12 16 20 240.1

1

10

100

3 mg

0.5 mg

1 mg

10 mg

N = 8/group

Mean + SEM

CR845 Demonstrated 16% Oral Bioavailability

21

PTH Phase II Study

Mean PTH Cmax Values for Subjects

Receiving Oral PTH(1-31)NH2 and Forsteo®

22

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

Me

an

% C

ha

ng

e L

S-B

MD

Phase III Study for Oral sCT:

Primary Endpoint (Change in LS BMD) Achieved

23

Small Molecule Experience

24

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

25

KANAMYCIN

PROJECT REVIEW

AMY STURMER

1.0 / 31JULY2013

Tigecycline

Case Study

26

• 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

27

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 LLC

ID PBS

ID 400mM CA, 26mM LLC

Time (hrs)

Pla

sma

Tig

ecyc

lin

e (m

cg/m

L)

Pla

sm

a C

on

ce

ntr

ati

on

(m

cg

/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 28

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

29

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)

30

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 4.0

0

30

60

90

120

150

180

15 mg

30 mg

45 mg

Time (hr)

Pla

sm

a T

igecycli

ne C

on

cen

trati

on

(ng

/mL

)

Pla

sm

a C

on

ce

ntr

ati

on

(n

g/m

L)

31

• 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

32

Synergy of CA and LLC

Demonstrated on Tigecycline

Mean %F

400mM CA/26mM LLC 21.64

0 CA/26mM LLC 8.86

(m)

33

Tigecycline Human PK Data

34

KANAMYCIN

PROJECT REVIEW

AMY STURMER

1.0 / 31JULY2013

Aminoglycoside (Kanamycin, Tobramycin)

Case Studies

35

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

36

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)

37

IV Tobramycin Study

38

Plasma Levels in 8 Dogs Given

Formulated Oral Tobramycin Capsules

39

Mean Plasma Levels in

Dogs Given Oral Tobramycin

40

(Adjusted for Tmax and Dose)

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)

%F

0.15

(0.08)

14.5

(2.7)

41

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)

42

Mean Pharmacokinetic Parameters for Kanamycin and Tobramycin

Following Oral Administration to Beagle Dogs (±SEM)

BCS Class III Molecule

Zanamivir Case Study

43

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

44

Zanamivir Dog Study Results

0

100

200

300

400

500

600

700

0 30 60 90 120 150 180 210 240

Zan

am

ivir

pla

sm

a c

on

cen

trati

on

(n

g/m

L)

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)

45

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

46

Small Molecule Experience

BCS Class II

47

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

48

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)

Fen

ofi

bra

te C

on

cen

trati

on

(mcg

/mL

)

Fenofibrate Equilibrium Solubility with

Increasing Concentrations of LLC

49

Safety

50

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

51

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

52

The Permeation

Enhancement by LLC is Transient

LeCluyse E.L., et al. (1993). J. Pharm. Exp. Therap. 265(2):955-962. 53

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

54

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

Modified from: Brayden, D. Permeation Enhancers and Oral Peptide Delivery.

Presented at the Roche Colorado Peptide Symposium, Sept. 12, 2011.

Modeling of

perturbed

membrane in

Caco-2 cells

indicates an

effective pore

radius ca. 20Å

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

• 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

59

LLC Regulatory and Clinical

60

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.

61

• 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

62

“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

63

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

64

Patents

65

Patents Summary

Oral Delivery Patents

• 7 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

66

Business Development

67

Working with Enteris

In-Vitro

• Solubility

• Permeability

In-Vivo

• Intra-duodenal:

Rat Study

In-Vivo

• Capsule: Dog

Study

In-Vitro & In-Vivo

• Stability; Design Space

• Tablet: Dog Study

Formulation

Development

Tablet Formulation Optimization

3 months 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

• 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

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

• 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.

74

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

75

E. coli Recombinant Peptide Manufacturing

Enteris’ Peptide Manufacturing Advantages

• Robust yield and purity

• Scalable

• Efficient

• Cost-effective

• Patented E. coli-based recombinant peptide

expression technology

• Peptide secreted directly into culture medium

• Enzymatic amidation of recombinant precursor

Direct Expression of Peptides Direct Expression of Peptides

76

Enteris’ Recombinant Manufacturing Process

77

kDa

66.2

45.0

31.0

21.5

14.4

6.5

sC

Tg

ly

Glu

co

se r

eg

ula

tory

pep

tid

e-g

ly

Glu

co

se r

eg

ula

tory

pep

tid

e a

nalo

g-g

ly

PT

H a

nalo

g-g

ly

SDS PAGE and HPLC Trace of Crude Medium

• Enriched Starting Material

• Extracellular yields of 400 to 1300 mg/L

• Reduced purification steps

High Yield and High Product Purity

at Low Cost

Recombinant Peptide Manufacturing

78

Amidated Peptide Manufacturing Example

rhPTH(1-34)NH2

0

100

200

300

400

500

600

700

800

1 5 9 13 17 21 25 29

Hours Post Induction

PT

H(1

- 34)

mg

/L

0.000

0.050

0.100

0.150

0.200

0.250

0.300

Feed

Rate

mL

/min

PTH(1-34)

mg/L

Feed Rate ml/min

0

100

200

300

400

500

600

700

800

1 5 9 13 17 21 25 29

Hours Post Induction

PT

H(1

- 34)

mg

/L

0.000

0.050

0.100

0.150

0.200

0.250

0.300

Feed

Rate

mL

/min

0

100

200

300

400

500

600

700

800

1 5 9 13 17 21 25 29

Hours Post Induction

PT

H(1

- 34)

mg

/L

0.000

0.050

0.100

0.150

0.200

0.250

0.300

Feed

Rate

mL

/min

0

100

200

300

400

500

600

700

800

1 5 9 13 17 21 25 29

Hours Post Induction

PT

H(1

- 34)

mg

/L

0.000

0.050

0.100

0.150

0.200

0.250

0.300

Feed

Rate

mL

/min

0

100

200

300

400

500

600

700

800

1 5 9 13 17 21 25 29

Hours Post Induction

PT

H(1

- 34)

mg

/L

0.000

0.050

0.100

0.150

0.200

0.250

0.300

Feed

Rate

mL

/min

0

100

200

300

400

500

600

700

800

1 5 9 13 17 21 25 29

Hours Post Induction

PT

H(1

- 34)

mg

/L

0.000

0.050

0.100

0.150

0.200

0.250

0.300

Feed

Rate

mL

/min

rhPTH(1-34)GLY Production and

Fermentation Feed Rate

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

The peptide: human PTH(1 - 34) - Gly

SVSEIQLMHNLGKHLNSMERVENLRKKLQDVHNFG MW = 4,116.8 Da

Step 1. Recombinant production of rhPTH(1-34)Gly35OH precursor

pPTH(1-34)G-03

7799 bp

PTH(1-34)gly

ompA

SEC-E LAC-IQ

KAN-R

PRLA-4

ompA

PTH(1-34)gly

P1

P2

P3

SecE P/O

P2

P1

Ori

Term

Term

Term

Term

79

Thank You