Design,Synthesis And Biological Evaluation Of Novel HIF-1 Inhibitors As Anticancer Agent

Mammalian cells need to maintain proper oxygen tension for aerobic metabolism,and hypoxia is a common feature of most solid tumors.Tumor cells under hypoxic conditions adapt to these hypoxic tension conditions by activating multiple survival pathways,of which hypoxia-inducible factor(HIF-1)is the most important pathway.Hypoxia-inducible factors regulate the transcriptional activation of more than one hundred genes by combining hypoxia response elements,thereby regulating the adaptation of tumor cells to hypoxia,such as vascular endothelial growth factor(VEGF),erythropoiesis(erythropoietin,EPO),Iron metabolism(transferrin),glucose transporters(GLUT-1,GLUT-3),insulin-like growth factor 2(IGF-2)and pH regulation(carbonic anhydrase 9,CA IX),etc.These genes are associated with blood vessels Generation,immune escape,metabolic reprogramming,growth factor signaling,invasion,tumor progression,and metastasis are closely related.A large number of clinical data indicate that in many cancers such as breast cancer,brain cancer,cervical cancer,colorectal cancer,acute lymphoma,myeloid leukemia,melanoma,gastric cancer,and liver cancer,lung cancer,ovarian cancer and pancreatic cancer increased HIF-1 protein levels are associated with increased radiotherapy and chemotherapy resistance,cancer progression,and patient mortality.At present,some marketed drugs have been found to have HIF-1 inhibitory effects,but there is no anti-tumor drug specifically targeting this target.In this paper,HIF-1 inhibitor YC-1 is used as the lead for structural optimization,its indazole core is retained,1-(3-(4-(trifluoromethoxy)phenyl)-1,2,4-oxadiazole moiety was added at the 1-position of indazole,and benzyl or piperidine containing different substituents moiety was added at the 3-position of indazole.These designed compounds TO1-TO11(Series A)and compounds TO12-TO20(Series B)was synthesized.Based on compound library in our laboratory,we designed a series of pyrazole derivatives containing covalent functional group,compounds TO21-TO36(Series C),by virtual screening and computer-aided drug desgn approach.The structures of all the synthesized compounds were confirmed by hydrogen spectrum(1H-NMR),carbon spectrum(13C-NMR)and mass spectrometry(MS).We conducted anti-tumor cells(mantle cell lymphoma cell line and liver cancer cell HepG2)proliferation experiments at the cell level for three series of compounds.The results showed that the growth inhibitory activity of TO1 and TO10 on ibrutinib(IBN)resistant cell lines Maver-1 and Z138 was significantly better than that of IBN;the compounds TO2 and TO3 had inhibitory activity on Maver-1 and Z138 better than IBN in IBN sensitive cell lines Jeko-1,Mino,Jeko-KO.Among liver cancer cells HepG2,TO1,TO2,TO3,TO6,and TO7 all showed better inhibitory activity,among which TO3 had the best inhibitory activity.TO-TO36 has also been tested on the MCL cell line.The experimental results show that TO21,TO28 have excellent inhibitory activity on MCL cells Z138,Rec-1,Jeko-1,Mino,Maver-1,and the activity is 10 times that of IBN;TO21,TO22 have antiproliferative activity comparable to IBN,and TO25 is comparable to IBN in Z138,Jeko-1,Mino,TO26,TO36 are comparable to IBN in Mino.Taking YC-1 as a control,the IC50 of compound TO1,TO3,and TO22 on HepG2 growth inhibition concentration was determined.The results showed that IC50 of TO3,TO1,TO22 were 9.39 pM,23.27 ?M and 24.49 ?M,respectively,better than YC-1(IC50=54.20 ?M).Western Blot analysis were conducted under both normoxic(20%O2)and hypoxic conditions(1%O2).The compounds TO1,TO3,and TO22 were compared with YC-1.It was found that the level of HIF-1? is more significantly increased in hypoxia than in normoxic condition.Under hypoxia,the level of HIF-1? was significantly reduced with the treatment of TO1,TO3,TO22 and YC-1.