Doramapimod

p38 MAPK:0.1 nM (Kd, cell free)

未经治疗的dTGRs的收缩压为204 mm Hg，但经过Doramapimod (30 mg/kg 每日) 处理后有所降低（166 mm Hg），而Sprague-Dawley大鼠保持正常血压。与dTGRs相比，Doramapimod处理后的心脏中β-肌球蛋白重链表达显著降低。Doramapimod的治疗显著减少了心脏纤维化、结缔组织生长因子、肿瘤坏死因子-α、白细胞介素-6和巨噬细胞浸润[4]。

Doramapimod (BIRB796) 是一种高效的 p38 MAPK 抑制剂（Kd：0.1 nM），能够阻断 LPS 刺激的 THP-1 细胞中 TNFα 的释放（IC50：18 nM）[1]。BIRB796 同时抑制 SAPK3/p38gamma 的活性及其激活，并阻断应激诱导的脚手架蛋白 SAP97 的磷酸化[2]。此外，BIRB 796 抑制了由 17-AAG 加 bortezomib 引起的 Hsp27 磷酸化，从而增强了细胞毒性。在骨髓基质细胞（BMSC）中，BIRB 796 抑制了由肿瘤坏死因子-α 或肿瘤生长因子-β1 触发的 p38 MAPK 的磷酸化和 IL-6 及血管内皮生长因子的分泌。BIRB 796 还抑制了 BMSCs 因黏附到 MM 细胞而诱导的 IL-6 分泌，从而抑制了肿瘤细胞增殖[3]。

DMSO:20 mg/mL (37.9 mM);Ethanol:26.4 mg/mL (50 mM)

Human embryonic kidney (HEK) 293 and HeLa cells were cultured in Dulbecco's modified Eagle's medium at 37 °C, supplemented with 10% fetal calf serum, 50 units of penicillin/ml, 50 μg/ml streptomycin, and 2 mM glutamine. Mouse embryonic fibroblasts were cultured as described previously, and C2C12 myoblasts were cultured. Cells were exposed to 0.5 M sorbitol for 30 min or 100 ng/ml EGF for 10 min and then lysed in buffer A (50 mM Tris-HCl, pH 7.5, 1 mM EGTA, 1 mM EDTA, 1 mM sodium orthovanadate, 10 mM sodium fluoride, 50 mM sodium β-glycerophosphate, 5 mM pyrophosphate, 0.27 M sucrose, 0.1 mM phenylmethylsulfonyl fluoride, 1% (v/v) Triton X-100) plus 0.1% (v/v) 2-mercaptoethanol and Complete proteinase inhibitor mixture from Roche Applied Science. Lysates were centrifuged at 18,000 × g for 5 min at 4 °C, and the supernatants were removed, quick-frozen in liquid nitrogen, and stored at –20 °C until use. When required, cells were preincubated for 1 h without or with 10 μM SB 203580 or 10 μM PD 184352 or with different concentrations of BIRB796 for the times indicated in the figures [2].

We studied male transgenic dTGRs and age-matched nontransgenic Sprague–Dawley (SD) rats (MDC). Local authorities approved the studies, and American Physiological Society guidelines for animal care were followed. We performed 2 different protocols. In protocol 2, untreated dTGR (n=15), dTGR+BIRB796 (30 mg/kg per day in the diet for 3 weeks; n=11), and SD (n=8 each group) rats were analyzed. Systolic blood pressure was measured weekly by tail cuff. Twenty-four– hour urine samples were collected in metabolic cages from weeks 5 to 7. Serum was collected at week 7. Serum creatinine and cystatin C were measured by clinical routine assays. Urinary rat albumin was determined by enzyme-linked immunosorbent assay. The aim of protocol 2 was to focus on electrophysiological alterations and mortality. Untreated dTGR (n=10), dTGR+BIRB796 (n=10), and SD (n=10) rats were studied up to week 8. Cardiac magnetic field mapping (CMFM) was performed at week 7 under isoflurane anesthesia. Echocardiography was performed as described earlier [4].