Synthesis, design, and antiproliferative evaluation of 6-(N-Substituted-methyl)pyrazolo[3,4-d]pyrimidines as the potent anti-leukemia agents

Cheng-Yen Chung; Sin-Min Li; Wei-Zheng Zeng; Naoto Uramaru; Guan-Jhong Huang; Shin-Hun Juang; Fung Fuh Wong

doi:10.1016/j.bioorg.2024.107424

Synthesis, design, and antiproliferative evaluation of 6-(N-Substituted-methyl)pyrazolo[3,4-d]pyrimidines as the potent anti-leukemia agents

Bioorg Chem. 2024 May 3:148:107424. doi: 10.1016/j.bioorg.2024.107424. Online ahead of print.

Authors

Cheng-Yen Chung¹, Sin-Min Li², Wei-Zheng Zeng³, Naoto Uramaru⁴, Guan-Jhong Huang⁵, Shin-Hun Juang⁶, Fung Fuh Wong⁷

Affiliations

¹ Department of Chinese Pharmaceutical Sciences and Chinese Medicine Resources, China Medical University, No. 91 Hsueh-Shih Rd., Taichung 40402, Taiwan.
² Institute of Translation Medicine and New Drug Development, China Medical University, No. 91 Hsueh-Shih Rd., Taichung 40402, Taiwan.
³ Institute of Nutrition, China Medical University, No. 100, Jingmao 1st Rd., Beitun Dist., Taichung 406040, Taiwan.
⁴ Department of Environmental Science, Nihon Pharmaceutical University, Komuro Inamachi Kita-adachi-gun, Saitama-ken 10281, Japan.
⁵ Department of Chinese Pharmaceutical Sciences and Chinese Medicine Resources, China Medical University, No. 91 Hsueh-Shih Rd., Taichung 40402, Taiwan; Department of Food Nutrition and Healthy Biotechnology, Asia University, No. 500, Lioufeng Rd., Wufeng, Taichung 41354, Taiwan.
⁶ School of Pharmacy, China Medical University, No. 100, Jingmao 1st Rd., Beitun Dist., Taichung 406040, Taiwan.
⁷ School of Pharmacy, China Medical University, No. 100, Jingmao 1st Rd., Beitun Dist., Taichung 406040, Taiwan. Electronic address: ffwong@mail.cmu.edu.tw.

PMID: 38728908
DOI: 10.1016/j.bioorg.2024.107424

Abstract

Pyrazolopyrimidine derivatives, including pyrazolopyrimidines, 6-aminopyrazolopyrimidines, 6-[(formyloxy)methyl]pyrazolopyrimidines, 6-(hydroxymethyl)pyrazolopyrimidine, and 6-(aminomethyl)pyrazolopyrimidines have been successfully prepared and tested against NCI-H226, NPC-TW01, and Jurkat cancer cell lines. Among the tested pyrazolopyrimidine compounds, we found 6-aminopyrazolopyrimidines and 6-(aminomethyl)pyrazolopyrimidines with essential o-ClPh or p-ClPh substituted moieties on N-1 pyrazole ring exhibited the best IC₅₀ inhibition activity for Jurkat cells. Furthermore, optimization of the SAR study on the C-6 position of pyrazolopyrimidine ring demonstrated that 6-(N-substituted-methyl)pyrazolopyrimidines 17b, 17d, and 19d possessed the significant IC₅₀ inhibitory activity for the different leukemia cell lines, especially for Jurkat, K-562, and HL-60. On the other hand, further SAR inhibition and docking model studies revealed that compound 19d, which has a 3-(1H-imidazol-1-yl)propan-1-amino side-chain on the C-6 position, was able to form four hydrogen bonds with residues Ala226, Leu152, and Glu194 and specifically extended into the P1 pocket subsite with Aurora A, resulting in improved inhibitory activity almost similar to SNS-314. To explore the anti-cancer mechanism, compound 19d was measured by Western blot analysis in Jurkat T-cells, however, it showed non-responsibility to Aurora B. For the further structural modifications on the lateral chain of compound 19d, compounds 24 with longer lateral chain were designed and synthesized for testing leukemia cell lines. However, compounds 24 was significantly decrease inhibition potency against leukemia cell lines. Based on the in-vitro results, compounds 17b and 19d could be considered to be the best potential lead drug in our study for the development of new and effective therapies for leukemia treatment. On the other hand, the DHFR inhibition results indicated compound 19d possessed good inhibitory activity and better than the reported naphthalene derivative. Through further comparisons of the model superposition of three-dimensional (3D) conformations in DHFR, compound 19d presented a similar structural alignment to Methotrexate and the reported naphthalene derivative and led to similar drug-like functional relationships. As a results, compound 19d would be a potential DHFR inhibitor for anti-leukemia drug candidate.

Keywords: 5-Aminopyrazoles; Antiproliferative; Jurkat; Leukemia; Pyrazolopyrimidines.