Rheumatoid Arthritis COLLAGEN-INDUCED ARTHRITIS: RAT THERAPEUTIC.

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Rheumatoid arthritis (RA) is an autoimmune disease associated with chronic inflammation, primarily in the joints. RA is characterized by synovial inflammation, swelling and autoantibody production, which result in the destruction of cartilage and bone in the small joints of hands, wrists and feet. Vium offers a well-established rat collagen-induced arthritis (CIA) model against which to validate the efficacy of potential therapeutic compounds1-2. In the rat CIA model, both B and T cells mediate joint inflammation and destruction with signature increases in the cytokines IL-1, IL-6, IL-17, and TNF-αwhich are validated targets for development for new therapeutics. VIUM METRICS Vium Arthritis IndexTM: The Arthritis Index provides an objective measure of function-related disease severity. The Vium index also increases the number of subjects or arms that can be simultaneously evaluated for efficacy by reducing labor-intensive measurements that require the same technician for consistency. Breathing Rate: Changes in breathing rate may be the result of disease processes or may occur following dosing of some therapeutics. Frequent assessment of breathing rates allows researchers to compare breathing rates across the course of the disease as well as focus on changes before and after therapeutic interventions. EXPERIMENTAL OVERVIEW The rat CIA model was conducted according to standard protocols3-5. Briefly, Lewis rats are inoculated with porcine collagen type II (2 mg/mL) in Incomplete Freund’s Adjuvant (IFA). Control animals are induced with IFA only. On study day 0, rats are given 200 μl intradermal injections on the back at 2 sites, and a single 100 μl booster injection on day 7. Disease onset occurs between 11 to 15 days with average day of onset occurring between study day 12 & 13. Our induction rate is > 95%. THERAPEUTIC TREATMENT REGIME Randomization: Rats are randomized into groups using motion prior to induction and disease severity at time of onset. Dosing: Rats are enrolled into treatment groups following a significant Vium Arthritis IndexTM score or an arthritis score of < 1. A standard study may dose rats for 14 – 28 days. Analysis: Treatment efficacy is assessed using Vium metrics (and option conventional measures) to compare:

• Disease severity between vehicle and treated rats Plots and statistical analysis are available on Vium Analytics Studio.

Vium metrics included with each study:

• Vium Arthritis IndexTM • Breathing rate

Optional conventional measures:

• Arthritis score • Ankle joint size (caliper) • Ankle joint histopathology

Vium Inc. | 411 Borel Avenue, Suite 616, San Mateo, CA 94402 650.797.2000 | info@vium.com | vium.com Version # 122-22

Positive control therapeutics. Several positive control therapeutics can be used including: • Methotrexate (daily, PO) • Enbrel (q3d, SC) • Dexamethasone (daily, PO or IP)

REFERENCES 1. Hegen M, Keith JC Jr, Collins M, Nickerson-Nutter CL. (2008) Utility of animal models for identification of potential

thereapeutics for rheumatoid arthritis. Ann Rheum Dis. 67: 1505-15. 2. Bevaart L, Vervoordeldonk MJ, Tak PP. (2010) Evaluation of therapeutic targets in animal models of arthritis: how does it relate

to rheumatoid arthritis? Arthritis Rheum. 62: 2192-205. 3. Benedele A. (2001) Animal models of rheumatoid arthritis. J Muscoloskelet Neuronal Interact. 1: 377-85. 4. Trentham DE, Townes AS, Kang AH. (1977) Autoimmunity to type II collagen an experimental model of arthritis. J Exp Med. 146:

857-68. 5. Courtenay JS, Dallman MJ, Dayan AD, Martin A, Mosedale B. (1980) Immunisation against heterologous type II collagen induces

arthritis in mice. Nature 283: 666-8.