The SMARTag TM ADC Technology Platform

Transcription

1 The SMARTag TM ADC Technology Platform

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5 World Class Protein Production Capability New State of the Art Facility Madison, WI Expanded GPEx Cell Line Engineering capacity 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 5

6 The Catalent-Redwood Advantage + 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 Our Value Proposition: Enable Biologics Innovators to Develop Better Antibody Drug Conjugates Faster 6

7 SMARTag Technology: Better Proteins Advanced Technology for the Development of Optimized Antibody Drug Conjugates: 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 SPECIFIC MODIFIABLE ALDEHYDE RECOMBINANT TAG 7

8 Limitations of Conventional ADC Technology 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 *Junutula, JR. et al. Nat. Biotechnol. 2008, 26, *Junutula, JR. et al. Clinical Cancer Res. 2010, 16, *Boswell, CA. et al. Bioconjugate Chemistry. 2011, 22,

9 SMARTag Technology: Next- Generation Antibody-Drug Conjugates Leveraging enzymes for site specific chemoenzymatic modification of proteins

10 Creation of Chemical Handle to Enable Site Specific Modification in Any Protein FGE recognition site (5 aa) N-terminus CxPxR C-terminus Protein of interest Formylglycine generating enzyme (FGE) cysteine Aldehyde bearing formylglycine FGE highly selective for Cys in CxPxR sequence to generate aldehyde tag (SMARTag TM ) Carrico IS, Carlson BL, and Bertozzi CR, Nat Chem Biol, 2007, 3, Wu, P, et al., Proc Natl Acad Sci U S A, 2009, 106, Hudak, J.E., et al., Angew Chem Int Ed Engl. 2012, 51, Rabuka, D., et al., Nat Protoc. 2012, 7,

11 Simple and Efficient Chemoenzymatic Approach for Generating Uniform Antibody Drug Conjugates Antibody is produced in cell line over-expressing FGE plasmid FGE recognition sequence inserted into antibody using standard cloning techniques Site-Specific Payload Placement Aldehyde tagged mabs purified using standard protocols mab is conjugated with proprietary cytotoxin-linkers that react selectively with aldheyde tags 11

12 SMARTag TM Technology: Optimized ADCs Homogeneous Drug Product Consistent cytotoxin/mab ratio: Batch to Batch Reproducibility ADC Structure Activity Relationship ( SAR ) Programmable Drug Placement: Optimized Therapeutic ADC SAR Index Controlled Drug Loading: Optimized Potency 2, 4 or 6 Payloads/Antibody 12

13 The SMARTag TM Technology Platform Includes All the Required Components to Assemble ADCs CYTOTOXIN Access to multiple Payloads Through SMARTag TM Approach LINKER Both Cleavable and Non-Cleavable Linkers ATTACHMENT Proprietary Conjugation Chemistries TAGGED ANTIBODY Site specific protein modification 13

14 Optimized ADCs: Site Specific Programmability Aldehyde Tags can be Placed Throughout the Protein 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 14

15 Aldehyde Tag Enables Advanced Conjugation Chemistry cytotoxin/linker 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 the SMARTag platform: maytansine, auristatin, PBD, duocarmycin >50 ADC linker/payload combinations successfully tested against multiple targets 15

16 Novel Conjugation Technology Provides Enhanced Stability: C-C Bond Formation Using Proprietary HIPS Chemistry N H NH 2 + R O H N H N H R N H NH R Pictet-Spengler reaction N H Me HN N Me + R O H R N H N Me N Me N H R N Me N Me Pictet-Spengler ligation Me HN N Me N Bioconjugation: C-C bond formation Me Me N N Me N Me N O H H O N N Agarwal, P.W. et al. Bioconjugate Chem 2013, 24,

17 Optimized ADCs: Homogeneity and Simplified Analytics ADCs generated using SMARTag Technology: D2 SMARTag site-specific system produces homogeneous ADCs D1 ADCs generated using conventional technology: D0 D8 Conventional cysteine conjugation results in 0-8 drugs per antibody; lysine conjugation has even greater heterogenicity Analytical HIC Profile of ADCs 17

18 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 18

19 SMARTag TM HER2 ADCs with Varied Payload Placement Position are Stable in Human Plasma After 14 Days Minimal loss of payloads (including cytotoxins) on SMARTag TM HER2 ADCs in human plasma at 37 o C Multiple payload placements are stable 19

20 Potent in vitro Cytotoxicity of SMARTag TM HER2 ADCs Generated with Redwood s Linker Library varied linker IC 50 s: TDM-1 = 250 pm αher2-hipslinker 1 ADC = 181 pm αher2-hipslinker 2 ADC = 219 pm αher2-hipslinker 3 ADC = 161 pm Isotype-HIPSLinker 1 ADC SMARTag TM HER2 ADCs exhibit potent cell killing activity SMARTag TM non-binding ADC control exhibited no cell killing over 6 days 20

21 Potent in vitro Cytotoxicity of SMARTag TM HER2 ADCs With Varied Payloads and Linkers Cytotoxicity of αher2 Maytansine or MMAE ADCs with cleavable linkers in NCI-N87 cells Cytotoxicity of αher2 MMAF with noncleavable linkers in NCI-N87 cells Free drug αher2-hipslinker1may ADC = 372 pm Isotype-HIPSLinker1May ADC αher2-hipslinker1auri1 ADC = 410 pm Isotype-HIPSLinker1Auri1 ADC Free drug αher2-hipslinker2auri2 ADC = 174 pm Isotype-HIPSLinker2Auri2 ADC SMARTag TM HER2 ADCs exhibit potent cell killing activity with different linkers/payloads SMARTag TM non-binding ADC controls exhibited no cell killing over 6 days 21

22 Tag Placement Does Not Impact In Vitro Cytotoxicity Against NCI-N87 Cells Heavy Chain C-Terminal Heavy Chain CH1 Light Chain IC 50 s: T-DM1 = 165 pm αher2-ct-hipslinker1may = 176 pm αher2-ch1-hipslinker1may = 150 pm αher2-lc-hipslinker1may = 97.8 pm varied placement SMARTag TM HER2 ADCs as potent as T-DM1 All payload placements active 22

23 SMARTag TM Her2 ADCs Control Tumor Growth Comparable to or Better than T-DM1 with Half the Drug Loading NCI-N87 Xenografts Panel of 7 CT-conjugated ADCs bearing different linkers (single dose 5 mg/kg) 23

24 Dose-Response Relationship Between SMARTag TM ADC Dose and Tumor Volume NCI-N87 Xenografts Dose-response study using a CT-conjugated ADC at 5 or 10 mg/kg 24

25 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) 25

26 SMARTag TM HER2 ADC Demonstrates Half-life Equivalent to Naked Antibody in Mouse PK Study Nominal Dose (mg/kg) T max (day) Half-life (day) Anti-HER2 Tagged Anti-HER2 Anti-HER2-ADC All 3 groups show similar PK parameters 26

27 Advantages vs. Other Site-Specific Systems 27

28 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/ , PRV Filed 1/11 Aldehyde-Tagged Immunoglobulin Polypeptides and Methods of Use Thereof - Application published Expanded FGE recognition sequences US2011/ , PRV Filed 3/08 Aldehyde Tags, Uses Thereof in Site-Specific Protein Modification - Application published Expanded use of aldehyde tags US2010/ , PRV Filed 2/09 Aldehyde-Tagged Protein-Based Drug Carriers and Methods of Use - Application published 28

29 References Publications Carrico, I.S., B.L. Carlson, and C.R. Bertozzi, Introducing genetically encoded aldehydes into proteins. Nat Chem Biol, (6): p Wu, P., et al., Site-specific chemical modification of recombinant proteins produced in mammalian cells by using the genetically encoded aldehyde tag. Proc Natl Acad Sci U S A, (9): p Rabuka, D., Chemoenzymatic methods for site-specific protein modification. Curr Opin Chem Biol (6): p Hudak, J.E., et al., Synthesis of heterobifunctional protein fusions using copperfree click chemistry and the aldehyde tag. Angew Chem Int Ed Engl (17): p Rabuka, D., et al., Site-specific chemical protein conjugation using genetically encoded aldehyde tags. Nat Protoc (6): p 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

30 To learn more about this technology or to request an expert consultation call REDWOOD BIOSCIENCE 5703 HOLLIS STREET EMERYVILLE, CA CATALENT PHARMA SOLUTIONS 14 SCHOOLHOUSE ROAD SOMERSET, NJ