Guru Bhavimani, Tamrakar Akhilesh K, Mandal Subhankar P, Kumar Prashantha B R, Sharma Aditya, Manjula Santhepete Nanjundaiah
Department of Pharmacology, JSS College of Pharmacy, JSS Academy of Higher Education & Research, Mysuru, India.
Division of Biochemistry and Structural Biology, CSIR-Central Drug Research Institute, Lucknow, India.
Pharmacology. 2022;107(1-2):90-101. doi: 10.1159/000519331. Epub 2021 Nov 4.
Peroxisome proliferator-activated receptor gamma (PPARγ) agonists are highly effective in treating insulin resistance. However, associated side effects such as weight gain due to increase in adipogenesis and lipogenesis hinder their clinical use. The aim of the study was to design and synthesize novel partial PPARγ agonists with weaker lipogenic effect in adipocytes and enhanced glucose transporter 4 (GLUT4) translocation stimulatory effect in skeletal muscle cells.
Novel partial PPARγ agonists (GS1, GS2, and GS3) were designed and screened to predict their binding interactions with PPARγ by molecular docking. The stability of the docked ligand-PPARγ complex was studied by molecular dynamics (MD) simulation. The cytotoxicity of synthesized compounds was tested in 3T3-L1 adipocytes and L6 myoblasts by MTT assay. The lipogenic effect was investigated in 3T3-L1 adipocytes using oil red O staining and GLUT4 translocation stimulatory effect in L6-GLUT4myc myotubes by an antibody-coupled colorimetric assay.
The molecular docking showed the binding interactions between designed agonists and PPARγ. MD simulation demonstrated good stability between the GS2-PPARγ complex. GS2 and GS3 did not show any significant effect on cell viability up to 80 or 100 μM concentration. Pioglitazone treatment significantly increased intracellular lipid accumulation in adipocytes compared to control. However, this effect was significantly less in GS2- and GS3-treated conditions compared to pioglitazone at 10 μM concentration, indicating weaker lipogenic effect. Furthermore, GS2 significantly stimulated GLUT4 translocation to the plasma membrane in a dose-dependent manner via the AMPK-dependent signaling pathway in skeletal muscle cells.
GS2 may be a promising therapeutic agent for the treatment of insulin resistance and type 2 diabetes mellitus without adiposity.
过氧化物酶体增殖物激活受体γ（PPARγ）激动剂在治疗胰岛素抵抗方面非常有效。然而，诸如因脂肪生成和脂质生成增加导致体重增加等相关副作用阻碍了它们的临床应用。本研究的目的是设计和合成新型的部分PPARγ激动剂，使其在脂肪细胞中具有较弱的脂质生成作用，并在骨骼肌细胞中增强葡萄糖转运蛋白4（GLUT4）转位刺激作用。
设计并筛选新型的部分PPARγ激动剂（GS1、GS2和GS3），通过分子对接预测它们与PPARγ的结合相互作用。通过分子动力学（MD）模拟研究对接的配体-PPARγ复合物的稳定性。采用MTT法在3T3-L1脂肪细胞和L6成肌细胞中测试合成化合物的细胞毒性。在3T3-L1脂肪细胞中使用油红O染色研究脂质生成作用，并通过抗体偶联比色法在L6-GLUT4myc肌管中研究GLUT4转位刺激作用。
分子对接显示了设计的激动剂与PPARγ之间的结合相互作用。MD模拟证明了GS2-PPARγ复合物之间具有良好的稳定性。在浓度高达80或100μM时，GS2和GS3对细胞活力没有任何显著影响。与对照组相比，吡格列酮处理显著增加了脂肪细胞内的脂质积累。然而，在10μM浓度下，与吡格列酮相比，GS2和GS3处理条件下的这种作用明显较小，表明脂质生成作用较弱。此外，GS2通过骨骼肌细胞中依赖AMPK的信号通路以剂量依赖的方式显著刺激GLUT4向质膜转位。
GS2可能是一种有前景的治疗药物，可用于治疗胰岛素抵抗和2型糖尿病且不会导致肥胖。
