LIFR/GPBAR1 modulator 1 is an orally active, potent GPBAR1 agonist (EC50= 0.2 μM) and LIFR inhibitor (IC50= 7.9 μM). It enhances the expression of leukemia inhibitory factor (LIF)-mediated LIFR and GPBAR1 mRNA, while significantly reducing the expression of fibrosis markers (COL1A1, ASMA, and TGFβ), decreasing TIMP1 levels, and increasing MMP9 expression. LIFR/GPBAR1 modulator 1 is applicable for research into human fibrotic diseases.

LIFR/GPBAR1 modulator 1 (Compound 2o) exhibits an 89.6% inhibition rate of LIFR at 10 μM concentration in HepG2 cells (IC50 = 7.9 μM), whereas in HEK293T cells, it activates GPBAR1 at a rate of 79.4% at the same concentration (EC50 = 0.2 μM). At concentrations ranging from 1 to 10 μM over 24 hours, it regulates the expression of fibrosis-related genes in a concentration-dependent manner in LX2 human hepatic stellate cells mediated by the leukemia inhibitory factor (LIF). At 10 μM, the compound upregulates mRNA expression of LIFR and GPBAR1 while significantly reducing the expression of profibrotic markers (COL1A1, ASMA, and TGFβ). It simultaneously decreases TIMP1 expression and increases MMP9 expression, indicating promotion of extracellular matrix (ECM) degradation.

Compound LIFR/GPBAR1 modulator 1 (Compound 2o) administered orally at a dose of 10 mg/kg once daily for 7 days can effectively reverse acute liver fibrosis induced by Carbon tetrachloride (CCl4) in mice. This reversal is evidenced by reduced hepatocyte injury, inhibition of inflammatory responses, decreased extracellular matrix (ECM) deposition, and downregulation of pro-fibrotic gene expression.