P164 MRT-6160, a VAV1-directed molecular glue degrader, inhibits disease progression and inflammation in a T-cell transfer model of Colitis

Abstract

Abstract Background VAV1 is an immune-restricted guanine nucleotide exchange factor critical for T-cell receptor (TCR) and B-cell receptor (BCR) signalling. The role of VAV1 in T-cells has been demonstrated in knockout mice, which exhibit decreased effector functions and resistance to autoimmune disease models, and in CRISPR-based screens, where VAV1 has been highlighted as a top hit among positive regulators of T-cell function. Until now, VAV1 has remained undruggable by conventional small molecules. MRT-6160 is a first-in-class molecular glue degrader that specifically targets VAV1 for proteasomal degradation. Given the role of VAV1 in TCR and BCR signalling, MRT-6160-mediated degradation could be an effective therapeutic approach to treating autoimmune disease. Methods We first tested the impact of MRT-6160-mediated human (h)VAV1 degradation on primary human cells using in vitro stimulation-based assays. Oral bioavailability of MRT-6160 and in vivo degradation of murine (m)VAV1 was then examined in mice. We tested the efficacy of MRT-6160 using a T-cell adoptive transfer model of colitis treating recipients of pathogenic CD45RBhigh CD4+ T-cells (8 per group) with anti-TNF (25 mg/kg, Q3D) or MRT-6160 (1 mg/kg, QD) for 41 days following cell transfer. Disease activity index (DAI), comprising stool consistency and weight loss, was assessed every 3 days. At study termination, mesenteric lymph nodes (mLN) were profiled by flow cytometry and colon tissues were assessed for cytokine expression and histopathology. Results Degradation of hVAV1 by MRT-6160 inhibited T-cell activation, proliferation, and cytokine production in a dose-dependent manner. Orally administered MRT-6160 degraded mVAV1 and inhibited colitis disease progression by 85%, reducing average end point DAI scores more than a standard of care control (vehicle, p<0.0001; anti-TNF, p=0.0107). Compared to vehicle treated groups, MRT-6160 treated mice had fewer IL-17A+ (12.6 and 5.3% respectively) and TNF+ (37.6 and 17% respectively) CD4+ T cells in mLN tissue. Colon weight:length ratio was significantly reduced in MRT-6160-treated mice (42.5 mg/cm) compared to vehicle (70.3 mg/cm) and anti-TNF (61.2 mg/cm). Finally, pro-inflammatory cytokine expression in the colon mucosa was reduced in MRT-6160-treated mice compared to vehicle (IL-6: 1305.8 and 682.3 fg/mg; TNF: 175.5 and 66.9 ng/mg; respectively), as well as histopathological evidence of disease. Conclusion MRT-6160 demonstrates strong activity in a preclinical model of colitis reducing DAI, effector cytokine production, and colon tissue damage. These data suggest that MRT-6160-mediated degradation of VAV1 may have therapeutic benefit in IBD patients and warrants further clinical development.