Figure 6.

Takinib reduces TNF in a murine LPS challenge. FVB mice treated i.p. with varying concentrations of Takinib, once (qw) or twice weekly (qbw), caused no significant adverse events or weight change over 8 weeks of chronic treatment (a). Bone marrow derived macrophages (BMDM’s) were plated at (2 × 10⁶/well) in a 24-well plate and treated with M-CSF (20 ng/mL) for 72 hours followed by a 24 hour rest period prior to activation with LPS (10 ng/mL) and IFNγ (50 ng/mL) and treatment with Takinib, at the indicated concentrations, or vehicle for 24 hours. Cytokine production was sampled from cell supernatant and examined by ELISA (b) (One-way ANOVA with Dunnett’s, n = 3± SEM). C57/bl6 male and female mice were sacrificed prior to LPS challenge and baseline TNF and IL-6 serum concentrations were examined by ELISA (c,d) (Student t-test, n = 3± SEM, 3 males, 3 females). C57/bl6 male and female mice were injected intraperitoneally (i.p.) with LPS (100 μg/kg in PBS) followed immediately by vehicle or Takinib (50 mg/kg in DMSO). 1 hour later, mice were sacrificed and blood samples clotted in centrifuge for 10 minutes at 3,000 rpm. Concentration of serum cytokines TNF and IL-6 determined by ELISA (c,d) (Student t-test, n = 5± SEM).