Anti-inflammatory Effect of Novel Spleen Tyrosine Kinase Inhibitor, SKI-O-592, on Fibroblast-like Synoviocyte in Rheumatoid Arthritis and THP-1 Cell

Seon Uk Kim¹, Hyun Jung Yoo ², Shin Eui Kang ³, Ji soo Park ⁴, Ra Ham Kim ⁵, Jung-Ho Kim ⁶, Hae-Jun Hwang ⁶, Jin Kyun Park ⁷, Eun Young Lee ⁷ and Yeong-Wook Song ⁸, ¹Department of Molecular Medicine and Biopharmaceutical Sciences, Graduate School of Convergence Science and Technology, and College of Medicine, Seoul National University, Seoul, Republic of Korea, ²Department of Molecular Medicine and Biopharmaceutical Sciences, Graduate School of Convergence Science and Technology and College of Medicine, Seoul National University, Seoul, Repulbic of Korea, Seoul, Republic of Korea, ³Department of Molecular Medicine and Biopharmaceutical Sciences, Graduate School of Convergence Science and Technology and College of Medicine, Seoul National University, Seoul, Republic of Korea, Seoul, Republic of Korea, ⁴Division of Rheumatology, Department of Internal Medicine, Seoul National University Hospital, Seoul, Korea., Seoul, Republic of Korea, ⁵Department of Molecular Medicine and Biopharmaceutical Sciences, Graduate School of Convergence Science and Technology and College of Medicine, Seoul National University, Seoul, Republic of Korea, Seoul, Republic of Korea, ⁶Oscotec Inc., Seongnam, Republic of Korea, ⁷Division of Rheumatology, Department of Internal Medicine, Seoul National University College of Medicine, Seoul, Korea, seoul, Republic of Korea, ⁸Seoul National University Hospital, Seoul, Republic of Korea

Meeting: 2019 ACR/ARP Annual Meeting

Keywords: rheumatoid arthritis (RA) and tyrosine kinase inhibition

Session Information

Date: Tuesday, November 12, 2019

Session Title: RA – Etiology & Pathogenesis Poster II

Session Type: Poster Session (Tuesday)

Session Time: 9:00AM-11:00AM

Background/Purpose: Rheumatoid arthritis (RA) is a chronic inflammatory autoimmune disease characterized by bone and cartilage destruction with leukocyte infiltration and activation at synovial tissue. The fibroblast-like synoviocytes (FLS) have a central role in disease pathogenesis and in vitro FLS invasiveness which correlates with articular damage in RA patients. Spleen tyrosine kinase (SYK) is a non-receptor tyrosine kinase known to have a crucial role in immune receptor signaling. Recently, a number of studies have revealed that aberrant SYK activation is associated with diverse allergic disorders and antibody-mediated autoimmune diseases such as rheumatoid arthritis, asthma and allergic rhinitis. A novel small-molecule SYK inhibitor, SKI-O-592, was designed and synthesized. The aim of this study is to evaluate the inhibitory effect of SYK inhibitor on inflammation and migration in RA FLS and THP-1 cell.

Methods: Selectivity of SKI-O-592 was evaluated by KinaseProfiler platform consisting of 293 human purified kinases using ATP Km for each kinase. A series of concentrations of ATP ranged from 10 to 1,500 µM were simultaneously incubated with cell-free kinase reaction. FLS were isolated from synovial tissues of RA patients, and treated with SKI-O-592 under tumor necrosis factor alpha (TNF-α). FLS were stimulated with TNF-α for 48 hr after 1 hr treatment of SKI-O-592. After stimulation, cell viability were measured using CCK-8 assay. The levels of IL-6, CXCL10, MMP-3 and TNF-α were measured in culture supernatant of RA FLS and THP-1 cell by ELISA. Wound healing assay were performed to evaluate cell migration ability. The expression of α-tubulin, phosphorylated SYK and phosphoinositide 3-kinase (PI3K) were determined by Western blotting.

Results: SKI-O-592 inhibited the activity of recombinant human SYK enzymes with an IC₅₀ of 5.1 nM. The kinases with more than 75% inhibition by 0.1 μM SKI-O-592 were SYK, YES and ROCK-II. The number of migrated cell to wound region was decreased in SKI-O-592 treated RA FLS without the change in cell viability. SKI-O-592 reduced the levels of cytokine and chemokine secretion including IL-6, CXCL10 and MMP-3 in RA FLS. Secreted TNF-α levels were decreased in THP-1 cell by SKI-O-592. Phosphorylation of PI3K was decreased after 30min of SKI-O-592 treatment. The viability and proliferation of the cells were not affected.

Conclusion: The novel SYK inhibitor, SKI-O-592 decreased the production of pro-inflammatory cytokine, chemokine and metalloproteinase in RA FLS. SKI-O-592 decreased the phosphorylation of PI3K. SKI-O-592 may provide a new therapeutic strategy in RA patients.

Disclosure: S. Kim, None; H. Yoo, None; S. Kang, None; J. Park, None; R. Kim, None; J. Kim, None; H. Hwang, None; J. Park, None; E. Lee, None; Y. Song, Astellas Pharma, Inc., 9.

To cite this abstract in AMA style:

Kim S, Yoo H, Kang S, Park J, Kim R, Kim J, Hwang H, Park J, Lee E, Song Y. Anti-inflammatory Effect of Novel Spleen Tyrosine Kinase Inhibitor, SKI-O-592, on Fibroblast-like Synoviocyte in Rheumatoid Arthritis and THP-1 Cell [abstract]. Arthritis Rheumatol. 2019; 71 (suppl 10). https://acrabstracts.org/abstract/anti-inflammatory-effect-of-novel-spleen-tyrosine-kinase-inhibitor-ski-o-592-on-fibroblast-like-synoviocyte-in-rheumatoid-arthritis-and-thp-1-cell/. Accessed January 20, 2021.

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