Design, synthesis, and target identification of new hypoxia-inducible factor 1 (HIF-1) inhibitors containing 1-alkyl-1H-pyrazole-3-carboxamide moiety

• Published in: Bioorganic & medicinal chemistry 2021-09, Vol.46, p.116375-116375, Article 116375
• Main Authors: Sakai, Marina, Takahashi, Nobuaki, Ikeda, Hiroaki, Furutani, Yutaka, Higuchi, Shoko, Suzuki, Takehiro, Dohmae, Naoshi, Kobayashi, Sayaka, Harada, Hiroshi, Kojima, Soichi, Matsuura, Tomokazu, Hattori, Akira, Kakeya, Hideaki
• Format: Article
• Language: English
• Subjects: Antigens, Neoplasm - genetics; Antigens, Neoplasm - metabolism; Antineoplastic Agents - chemical synthesis; Antineoplastic Agents - chemistry; Antineoplastic Agents - pharmacology; Cancer; Carbonic Anhydrase IX - antagonists & inhibitors; Carbonic Anhydrase IX - genetics; Carbonic Anhydrase IX - metabolism; Cell Line, Tumor; Cell Proliferation - drug effects; Chemotherapy; Dose-Response Relationship, Drug; Drug Design; Drug Screening Assays, Antitumor; Female; Humans; Hypoxia-inducible factor (HIF); Hypoxia-Inducible Factor 1, alpha Subunit - antagonists & inhibitors; Hypoxia-Inducible Factor 1, alpha Subunit - metabolism; Molecular Structure; Pyrazoles - chemical synthesis; Pyrazoles - chemistry; Pyrazoles - pharmacology; Structure-Activity Relationship; Target identification; Vascular Endothelial Growth Factor A - antagonists & inhibitors; Vascular Endothelial Growth Factor A - genetics; Vascular Endothelial Growth Factor A - metabolism
• ISSN: 0968-0896; 1464-3391
• DOI: 10.1016/j.bmc.2021.116375

[Display omitted] •A promising HIF-1 inhibitor KUSC-5037 was developed during extensive SAR study.•A fluorescent probe and a bifunctional probe of KUSC-5037 were synthesized and utilized.•ATP5B, a β subunit of FoF1-ATP synthase, was identified as the target protein. Hypoxia-inducible factor 1 (HIF-1) is a promising drug target for cancer chemotherapy. In our screening program aimed at identifying new HIF-1 inhibitors by using a hypoxia-responsive luciferase reporter gene assay, KUSC-5001 containing the 1-alkyl-1H-pyrazole-3-carboxamide moiety was found as a potential hit molecule. During an extensive structure–activity relationship (SAR) study, we developed a more effective HIF-1 inhibitor KUSC-5037 (IC50 = 1.2 μM). Under hypoxic conditions, KUSC-5037 suppressed the HIF-1α (a regulatory subunit of HIF-1) mRNA, causing decreases in the gene expression of HIF-1 target genes such as carbonic anhydrase 9 (CA9) and vascular endothelial growth factor (VEGF) genes. Furthermore, by applying our fluorescent and bifunctional probes, ATP5B, a catalytic β subunit of mitochondrial FoF1-ATP synthase, was identified as a target protein of KUSC-5037. These results indicate that the derivatives of KUSC-5037 containing the 1-alkyl-1H-pyrazole-3-carboxamide moiety are promising lead molecules for the inhibition of HIF-1 signaling via FoF1-ATP synthase suppression.