CU-T12-9 is a potent TLR1/2 agonist(EC50 of 52.9 nM in HEK-Blue hTLR2 SEAP assay). It acts by activating the NFkB pathway, upregulating proinflammatory cytokines, and enhancing TLR1 and TLR2 dimerization.CU-T12-9 activates both the innate and the adaptive immune systems. CU-T12-9 selectively activates the TLR1/2 heterodimer, not TLR2/6. CU-T12-9 signals through NF-κB and invokes an elevation of the downstream effectors TNF-α, IL-10, and iNOS.

TLR1:52.9 nM （EC50）（in HEK-Blue cells), TLR2:52.9 nM （EC50）（in HEK-Blue cells)

CU-T12-9 directly targets TLR1/2 to initiate downstream signaling. It specifically induces TLR1/2 activation, which can be blocked by either the anti-hTLR1 or anti-hTLR2 antibody, but not the anti-hTLR6 antibody. By binding to both TLR1 and TLR2, CU-T12-9 facilitates TLR1/2 heterodimer formation, activating downstream signaling. Fluorescence anisotropy assays revealed competitive binding to the TLR1/2 complex between CU-T12-9 and Pam3CSK4, with an IC50 of 54.4 nM. CU-T12-9 signals through nuclear factor κB (NF-κB) and elevates downstream effectors tumor necrosis factor-α (TNF-α), interleukin-10 (IL-10), and inducible nitric oxide synthase (iNOS)[1].