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ML RR-S2 CDA is a synthetic cyclic dinucleotide (CDN) that contains non-canonical 2'5'-phosphodiester bonds and is an activator of stimulator of interferon genes (STING). It contains mixed linkages (ML) with both 2'5' and 3'5' linkages, which leads to increased thermal stability of human STING in a differential scanning fluorimetry (DSF) assay. ML RR-S2 CDA increases type I interferon production by THP-1 human monocytes relative to unmodified cyclic di-AMP , indicating the ML enhances its action at human STING. It induces expression of IFN-β and the pro-inflammatory cytokines TNF-α, IL-6, and Mcp-1 in murine bone marrow macrophages (BMM) isolated from wild-type, but not STING-/-, mice. ML RR-S2 CDA also induces IFN-β expression in peripheral blood mononuclear cells (PBMCs) isolated from donors carrying STINGWT/WT, STINGWT/REF, and STINGWT/HAQ alleles. In vivo, ML RR-S2 CDA initiates tumor regression and prevents tumor growth upon tumor cell reimplantation in 4T1 breast and CT26 colon cancer mouse xenograft models..
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