The SMARTagTM ADCTechnology Platform

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World Class Protein Production Capability

New State of the Art Facility – Madison, WI

• Expanded GPEx® Cell Line Engineeringcapacity

• Flexible Non cGMP production up to 250L and cGMP production scale up to 1000 L

• Latest single-use systems with skid-based processing

• Integrated development services to solve your most difficult biologic challenges

• Expanded + 100,000 square feet

Experience with aldehyde tagged antibody production and ability to produce at GMP scale

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The Catalent-Redwood Advantage

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Our Value Proposition: Enable Biologics Innovators to Develop Better

Antibody Drug Conjugates Faster

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Biologic Development Expertise

• Advanced GPEx® Expression System

• State-of-the-art Biomanufacturing and Integrated Analytical Services

• Best Practice Partnership Management

Advanced ADC Technology

• Proprietary Conjugation Platform

• Advanced Cytotoxin-Linker Chemistry

• ADC Development Expertise

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SMARTag™ Technology: Better Proteins

Site-Specific, Programmable Drug Placement • Novel patented “aldehyde tagging” technology• Control of drug location to maximize ADC performance• Improved consistency of ADC product• Better regulatory compliance

Proprietary Cytotoxin-linkers and Conjugation Chemistry• Superior linker stability• Library of cytotoxin/linkers to optimize efficacy by target• Commercial access to payloads through unique chemistry

Efficient and Scalable Process• One step in vivo tag generation• Compatible with any cell-based expression system• Simplified analytics

Advanced Technology for the Development of Optimized Antibody Drug Conjugates:

MODIFIABLE

SPECIFIC

RECOMBINANT

ALDEHYDE

TAG

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Limitations of Conventional ADC Technology

*Junutula, JR. et al. Nat. Biotechnol. 2008, 26, 925-932.*Junutula, JR. et al. Clinical Cancer Res. 2010, 16, 4769-4778.*Boswell, CA. et al. Bioconjugate Chemistry. 2011, 22, 1994-2004.

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1. Toxicity: aggregation & first pass metabolism in liver, linker instability*

2. Variable Potency: uncontrolled payload loading, PK liability*

3. CMC*: lot to lot reproducibility, complex manufacturing, challenging analytics

SMARTag™ Technology: Next-Generation Antibody-Drug Conjugates

Leveraging enzymes for site specificchemoenzymatic modification of proteins

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Creation of Chemical “Handle” to Enable Site Specific Modification in Any Protein

N-terminus C-terminus

Formylglycine generating enzyme(FGE)

Aldehyde bearing formylglycinecysteine

CxPxR

FGE highly selective for Cys in CxPxR sequence to generate aldehyde tag (SMARTagTM)

FGE recognition site (5 aa)

Protein of interest

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Carrico IS, Carlson BL, and Bertozzi CR, Nat Chem Biol, 2007, 3, 321-2.Wu, P, et al., Proc Natl Acad Sci U S A, 2009, 106, 3000-5.

Hudak, J.E., et al., Angew Chem Int Ed Engl. 2012, 51, 4161-5.Rabuka, D., et al., Nat Protoc. 2012, 7, 1052-67.

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plasmid

FGE recognition sequenceinserted into antibody usingstandard cloning techniques

Antibody is produced in cell line over-expressing FGE

Aldehyde tagged mAbspurified using standard

protocols

mAb is conjugated with proprietary cytotoxin-linkers that react selectively with

aldheyde tags

Site-Specific Payload Placement

Simple and Efficient Chemoenzymatic Approach for Generating Uniform Antibody Drug Conjugates

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SMARTagTM Technology: Optimized ADCs

Homogeneous Drug Product

Consistent cytotoxin/mAb ratio:

Batch to Batch Reproducibility

ADC Structure Activity Relationship (“SAR”)Programmable Drug

Placement:Optimized Therapeutic

IndexADC SARControlled Drug Loading:

Optimized Potency – 2, 4 or 6 Payloads/Antibody

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The SMARTagTM Technology Platform Includes All the Required Components to Assemble ADCs

CYTOTOXINAccess to multiple Payloads Through SMARTagTM Approach

LINKERBoth Cleavable and Non-Cleavable Linkers

ATTACHMENTProprietary Conjugation Chemistries

TAGGED ANTIBODYSite specific protein modification

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• Multiple tagged regions (in green)

• Biophysical properties

— Aggregation, >95% monomer

— Aldehyde tags can be introduced at multiple positions with

— No impact on antigen binding

— No impact on internalization

— No impact on melting temp

— No impact on FcRN binding

— No impact on protein expression

Optimized ADCs: Site Specific Programmability

Aldehyde Tags can be Placed Throughout the Protein

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cytotoxin/linker

Aldehyde Tag Enables Advanced Conjugation Chemistry

• Proprietary cleavable and non-cleavable linker library

• Advanced conjugation chemistry

• C-C ligation – enhanced plasma stability, half-life > 20 days

• Payloads from every commonly used ADC drug class provided on theSMARTag platform: maytansine, auristatin, PBD, duocarmycin

• >50 ADC linker/payload combinations successfully tested against multiple targets

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NH

NH2

R

O

H+

NH

N H

R NH

NH

R

NH

R

O

H+

NH

NN

NH

N MeHN

Me

NN

Me

Me

RR

Me

Me

O

H H

O N

NNMe Me

N

N NMeMe

N

N MeHN

Me

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Novel Conjugation Technology Provides Enhanced Stability:C-C Bond Formation Using Proprietary HIPS Chemistry

Pictet-Spengler reaction

Pictet-Spengler ligation

Bioconjugation:C-C bond formation

Agarwal, P.W. et al. Bioconjugate Chem 2013, 24, 846–851..

D1

D2

D0

D8

Analytical HIC Profile of ADCs

Optimized ADCs: Homogeneity and Simplified Analytics

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ADCs generated using SMARTag Technology:

ADCs generated using conventional technology:

SMARTag site-specific system produces homogeneous ADCs

Conventional cysteine conjugation results in 0-8 drugs per antibody; lysine conjugation has even greater heterogenicity

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Analytical Expertise to support ADCs

DAR: HIC, UV, RP-HPLC, MS

Drug distribution: HIC, MS, iCE

Purity/Impurity Characterization: SEC,

SEC/MALLS, SDS-PAGE or CE, MS

Charge heterogeneity: iCE, IEF, CEX, MS

Free drug (unconjugated): HPLC, ELISA, CE

Potency: ELISA, Cell based assays (binding and

cytotoxic)

LC/MS based method: Intact MW, subunit

analysis, peptide map, glycan profile

Biophysical: Fluorescence

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SMARTagTM HER2 ADCs with Varied Payload Placement Position are Stable in Human Plasma After 14 Days

•Minimal loss of payloads (including cytotoxins) on SMARTagTM HER2 ADCs in human plasma at 37oC

•Multiple payload placements are stable

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Potent in vitro Cytotoxicity of SMARTagTM HER2 ADCsGenerated with Redwood’s Linker Library

•SMARTagTM HER2 ADCs exhibit potent cell killing activity

•SMARTagTM non-binding ADC control exhibited no cell killing over 6 days20

IC50s: TDM-1 = 250 pMαHER2-HIPSLinker 1 ADC = 181 pMαHER2-HIPSLinker 2 ADC = 219 pMαHER2-HIPSLinker 3 ADC = 161 pMIsotype-HIPSLinker 1 ADC

varied linker

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Potent in vitro Cytotoxicity of SMARTagTM HER2 ADCsWith Varied Payloads and Linkers

Free drug αHER2-HIPSLinker1May ADC = 372 pM

Isotype-HIPSLinker1May ADC αHER2-HIPSLinker1Auri1 ADC = 410 pMIsotype-HIPSLinker1Auri1 ADC

Cytotoxicity of αHER2 Maytansine or MMAE ADCs with cleavable linkers in

NCI-N87 cells

Cytotoxicity of αHer2 MMAF with non-cleavable linkers in NCI-N87 cells

Free drugαHER2-HIPSLinker2Auri2 ADC = 174 pMIsotype-HIPSLinker2Auri2 ADC

•SMARTagTM HER2 ADCs exhibit potent cell killing activity with different linkers/payloads

•SMARTagTM non-binding ADC controls exhibited no cell killing over 6 days

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•SMARTagTM HER2 ADCs as potent as T-DM1

•All payload placements active

Tag Placement Does Not Impact In Vitro CytotoxicityAgainst NCI-N87 Cells

IC50s: T-DM1 = 165 pMαHER2-CT-HIPSLinker1May = 176 pMαHER2-CH1-HIPSLInker1May = 150 pMαHER2-LC-HIPSLinker1May = 97.8 pM

varied placement

Heavy Chain C-Terminal

Heavy Chain CH1

Light Chain

SMARTagTM Her2 ADCs Control Tumor Growth Comparable to or Better than T-DM1 with Half the Drug Loading

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NCI-N87 Xenografts

Panel of 7 CT-conjugated ADCs bearing different linkers (single dose 5 mg/kg)

Dose-Response Relationship BetweenSMARTagTM ADC Dose and Tumor Volume

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NCI-N87 Xenografts

Dose-response study using a CT-conjugated ADC at 5 or 10 mg/kg

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Specific Conjugation Sites Impact In Vivo Efficacy

NCI-N87 Xenografts

Panel of 3 ADCs conjugated with the same linker at 3 distinct locations (single dose 5 mg/kg)

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Nominal Dose (mg/kg) Tmax (day) Half-life (day)

Anti-HER25.0 0.25 10.9

Tagged Anti-HER25.0 0.25 7.8

Anti-HER2-ADC5.0 0.25 10.9

SMARTagTM HER2 ADC Demonstrates Half-life Equivalent to Naked Antibody in Mouse PK Study

All 3 groups show similar PK parameters

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Advantages vs. Other Site-Specific Systems

Redwood Public IP

• Use of FGE in bioconjugate creation/resulting composition, PRV Filed 9/06“Aldehyde Tags, Uses Thereof in Site-Specific Protein Modification”

→ U.S. method claims issued 7/11 (7,985,783) and 1/13 (8,349,910)→ U.S. composition-of-matter claims issued 1/12 (8,097,701)

• Tag placement and ADC generation – US2012/0183566, PRV Filed 1/11“Aldehyde-Tagged Immunoglobulin Polypeptides and Methods of Use Thereof”- Application published

• Expanded FGE recognition sequences – US2011/0117621, PRV Filed 3/08“Aldehyde Tags, Uses Thereof in Site-Specific Protein Modification”- Application published

• Expanded use of aldehyde tags – US2010/0210543, PRV Filed 2/09“Aldehyde-Tagged Protein-Based Drug Carriers and Methods of Use”- Application published

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References

PublicationsCarrico, I.S., B.L. Carlson, and C.R. Bertozzi, Introducing genetically encoded

aldehydes into proteins. Nat Chem Biol, 2007. 3(6): p. 321-2.Wu, P., et al., Site-specific chemical modification of recombinant proteins

produced in mammalian cells by using the genetically encoded aldehydetag. Proc Natl Acad Sci U S A, 2009. 106(9): p. 3000-5.

Rabuka, D., Chemoenzymatic methods for site-specific protein modification. CurrOpin Chem Biol. 2011 14(6): p. 790-6.

Hudak, J.E., et al., Synthesis of heterobifunctional protein fusions using copper-free click chemistry and the aldehyde tag. Angew Chem Int Ed Engl. 2012 51(17): p. 4161-5.

Rabuka, D., et al., Site-specific chemical protein conjugation using genetically encoded aldehyde tags. Nat Protoc. 2012 7(6): p. 1052-67.

Agarwal, P.W., et al., Hydrazino-Pictet-Spengler Ligation as a Biocompatible Method for the Generation of Stable Protein Conjugates. Bioconjugate Chem., 2013, 24 (6), p. 846–851.

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To learn more about this technology or to request an expert consultation call

CATALENT PHARMA SOLUTIONS14 SCHOOLHOUSE ROADSOMERSET, NJ 08873+ 1 866 720 3148www.catalent.com

REDWOOD BIOSCIENCE5703 HOLLIS STREETEMERYVILLE, CA 94608+ 1 510 343 6034www.redwoodbioscience.com