Discovery of a Small Molecule Inhibitor of the Wnt Pathway (SM04690) as a Potential Disease Modifying Treatment for Knee Osteoarthritis

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1 Discovery of a Small Molecule Inhibitor of the Wnt Pathway (SM469) as a Potential Disease Modifying Treatment for Knee Osteoarthritis Vishal Deshmukh, Ph.D., Charlene Barroga, Ph.D., Yong Hu, Ph.D., John Hood, Ph.D., and Yusuf Yazici, M.D.

2 DISCLOSURES Vishal Deshmukh, Ph.D. o Financial disclosure: Samumed, LLC; salary and equity Charlene Barroga, Ph.D. o Financial disclosure: Samumed, LLC; salary and equity Yong Hu, Ph.D. o Financial disclosure: Former employee of Samumed, LLC; equity John D. Hood, Ph.D. o Financial disclosure: Co-founder and former employee of Samumed, LLC Yusuf Yazici, M.D. o Financial disclosure: Samumed, LLC; salary and equity eular European League Against Rheumatism

3 Pathophysiology of Osteoarthritis Mechanical forces and inflammation result in degenerative tissue remodeling in OA. Induce cartilage catabolic enzymes - matrix metalloproteinases (MMPs), aggrecanases, etc. Cartilage loss and subchondral bone remodeling Healthy Osteoarthritis Bone marrow Synovial membrane Articular Cartilage Mesenchymal Stem Cells Bone Remodeling Bone Sclerosis Synovitis Inflammatory cells and cytokines Osteophyte Cartilage Loss Cartilage Degradation MMPs, ADAMTS Chondrocyte Hypertrophy Figure adapted from Bush J & Beier F. Nature Med. 213;19(6):667-9.

4 Wnt Pathway and Osteoarthritis Mechanical stress Cytokines Chondrocytes + Wnt activity Synovium Progenitor cells + Wnt activity - Osteoblasts Catabolic Enzymes Chondrocytes Normal Cartilage Cartilage Degradation The Wnt signaling pathway is involved in stem cell control and regeneration of tissues Increased Wnt signaling contributes to the pathophysiology of OA 1-2 Wnt signaling is involved in increased bone formation and cartilage breakdown Progenitor cells reside in the synovium and subchondral bone 3-5 Hypothesis: Inhibiting the Wnt Pathway protects and regenerates cartilage 1. Rudnicki JA & Brown AM. Dev Biol. 1997;185(1): Im GI, et al. Biotechnol Lett. 211;33(5): Thomas RS, et al. Arthritis Res Ther. 211;13(6):R Loughlin J. Curr Opin Rheumatol. 25;17(5): Blom AB, et al. Arthritis Rheum. 29;6(2): Figure adaptations: and Bush J & Beier F. Nature Med. 213;19(6):

5 Proposed Therapy: SM469 SM469 is a small molecule Wnt inhibitor in development for the treatment of OA SM469 demonstrated the following properties in pre-clinical studies: Decreased inflammation Decreased cartilage degradation Regenerated cartilage Sustained local exposure and no observable systemic toxicity 5

6 In vitro Efficacy- SM469

7 p g /m l Decreased inflammation: SM469 suppressed inflammatory cytokines Synovial fibroblasts IL1β, TNFα and IL-6 are associated with the pathophysiology of OA 1 Cellular assay: Synovial fibroblasts stimulated with IL1β and THP-1 monocytes stimulated with LPS to induce cytokine production p g /m l 4 TNFa 3 EC 5 ~ 35 nm Then treated with SM469 Cytokine production quantified by ELISA Dose dependent inhibition of both TNFa and IL-6 production demonstrated in both cell types IL-6 EC 5 ~ 24 nm Glyn-Jones S, et al. Lancet. 215;386(9991)

8 Decreased cartilage degradation: SM469 inhibited protease production Fold change (X/unstimulated) Fold change (X/unstimulated) Fold change (X/unstimulated) In OA, cytokines induce cartilage catabolic enzymes Increased Wnt signaling increases protease expression 1 Dose dependent inhibition of protease expression demonstrated Cellular assay human chondrocytes: Induce proteases Treat Measure TNFα + Oncostatin M SM469 or Control qpcr: MMP 1, 3, & MMP1 Control 469 1nM 469 3nM MMP3 Control 469 1nM 469 3nM MMP Control 469 1nM 469 3nM 1. Enochson L, et al. OA Cart. 214;22(4): p<.5 p<.1 8

9 Decreased cartilage degradation: SM469 inhibited GAG and Nitric Oxide release N O re le a s e (u M ) 8 Secreted GAG Glycosaminoglycan (GAG) are components of cartilage matrix Secreted/extracellular GAG = cartilage breakdown Inhibition of GAG and Nitric Oxide (NO) release demonstrated S e c re te d G A G (E x tra c e llu la r/in tra c e llu la r) Cellular assay human chondrocytes: TNFα + OM IL1β SM Induce catabolism Treat Measure IL1β or TNFα + Oncostatin M SM469 or Control Secreted GAG and NO Nitric Oxide p<.5 p<.1 p<.1 2 SM469 protected chondrocytes from catabolic breakdown TNFα + OM IL1β SM

10 Fold Change Over DMSO Regenerated cartilage: SM469 induced functional chondrogenesis GAG (ug/mg) s G A G (u g /m g ) SM469 (3nM) Control (DMSO) Fold Change Over DMSO 21 day cellular assay hmscs: Treated with SM469 every 7 days Cells stained for biomarkers and gene expression measured by qpcr Increased sulfated glycosaminoglycans (sgag) with SM469 treatment Functional chondrocytes-cartilage matrix synthesis Alcian Blue Safranin O Type II collagen 6 Chondrogenic Genes DMSO SM469 (3nM) 8 GAG 1. 5 Osteogenic Genes DMSO SM469 (3nM) S O X9 A g g re c a n C o l2 A TG F b 1 TIM P 1 DMSO SM469 (3nM). C o l1 A O s t e o c a lc in A L P L B M P 4 p<.5 p<.1 p<.1 1

11 In vivo Efficacy

12 S M (n M ) SM469 had sustained local exposure and no systemic toxicity 6 Rats (Sprague Dawley): Single intra-articular injection (.3ug) Compound is retained in joint above the target concentration level (~3 nm) for >6 months Compound is undetectable in plasma at all time points Expected therapeutic level (~3 nm) D a y s C a rtila g e B o n e P la s m a Intra-articular (IA) injection in Rats (Sprague Dawley) and Dogs (Beagle): Single or multiple (6 or 9 once-monthly) IA injections No systemic toxicity - body weight, target or non-target organ effects, ECG and clinical pathology at doses up to 4X the expected clinical dose 12

13 Decreased cartilage degradation: ACLT + pmmx model of OA R e la tiv e E x p re s s io n Rat ACLT + pmmx model- anterior cruciate ligament transection (ACLT) combined with partial medial meniscectomy (pmmx) Inject SM469 single dose, intra-articular after 1 week Rat knee analyzed 5 and 13 weeks post-surgery for OA cartilage pathology Week 5 qpcr evaluation of protease enzymes in cartilage Decreased protease expression in cartilage with SM469 treatment Vehicle SM469 (.3ug). M M P 1 M M P 3 M M P 1 3 A D A M T S 5 p<.5 p<.1 p<.1 N=12 rats/group SM469 protected cartilage from catabolic breakdown 13

14 R e la tiv e E x p re s s io n SM469 regenerated cartilage: ACLT + pmmx model of OA GAG (ug/g) qpcr evaluation of cartilage production markers Increased expression of cartilage markers with SM469 treatment Increased sulfated glycosaminoglycans (sgag) - cartilage matrix No change in Col1a (hypertrophic marker) Week ns Vehicle SM469 (.3ug) s G A G (u g /g ) sgagg A G Vehicle SM469 (.3ug) C o l2 a 1 C O M P A g g re c a n C o l1 a p<.5 p<.1 ns- not significant N=12 rats/group SM469 induced chondrocyte and cartilage matrix production 14

15 C O M P (n g /m l) SM469 regenerated cartilage: Improved OA biomarkers and OARSI scores P IIA N P (n g /m l) OARSI Score Decreased serum COMP and increased serum PIIANP observed with SM469 treatment Safranin O-stained sections from the rat knee scored (blinded) using OARSI system OARSI cartilage pathology score measures cartilage matrix loss, fissures and subchondral bone remodeling, based on stage and grade of cartilage damage COMP Vehicle SM469.3ug PIIANP Vehicle SM469.3ug 6 4 OARSI score Week W e e k 3 W e e k 6 W e e k 4 W e e k 5 Vehicle SM469.1 ug SM469.3 ug (.1µg) (.3µg) SM469 improved joint health p<.5 p<.1 N=12 rats/group 15

16 SM469 regenerated cartilage Safranin O-stained sections from the rat knee analyzed 13 weeks post-surgery for OA cartilage pathology Increased cartilage thickness and decreased fissures observed with a single intraarticular injection of SM469 femur Control Knee 12 weeks tibia Treated Knee (.3µg) 13 weeks Increased Cartilage SM469 increased cartilage thickness 16

17 Summary Wnt signaling is a critical pathway in osteoarthritis In preclinical models, SM469: Inhibited inflammatory cytokine and protease production Induced chondrogenesis Had sustained local availability and no systemic exposure Had no observable systemic toxicity Phase 1 clinical data Poster SAT428, Saturday June 11 th, 1:15-11:45 17

18 Thank you 18

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