Abstract
Novel inhibitors of GlcNAc-Ts have been reported based on benzoxazolone appended amino acids/esters which showed a competitive inhibition of the enzyme with IC50 values in lower micromolar range. The compounds 1b–5b showed a better inhibition profile against GlcNAc-Ts as compared to 1a–5a as evidenced by the in vitro 96 well plate ELISA experiment and the in vitro cytotoxicity studies on the cancer cell lines PDAC, Caco-2, MDA-MB-231, HT-29, PC-3, and A549. The execution of ELISA and cytotoxicity experiments on the compounds 1a–5a in the presence of esterase enzyme along with the appraisal of the lipophilicity index, and calculation of Ki and Ka values for the enzyme-inhibitor complex further indicated that the compounds 1a–5a serve as prodrugs of the compounds 1b–5b owing to a better lipophilicity index of the former. Double reciprocal plots were constructed for 2 different concentrations of the test compounds 1b–5b and that of the natural substrate of OGT. A competitive mode of inhibition was observed for the test molecules which was further confirmed by molecular docking analysis of these molecules in the active site of OGT in the presence of its natural inhibitor. Mechanistic evaluation of the inhibition of OGT by test molecules was performed by NMR and HPLC experiments where hydrolysis of exocyclic urea linkage in the test molecules was observed on their incubation with the enzyme over a period.
Graphical abstract
Similar content being viewed by others
References
Adelfinskaya O, Terrazas M, Froeyen M, Marlière P, Nauwelaerts K, Herdewijn P (2007) Polymerase-catalyzed synthesis of DNA from phosphoramidate conjugates of deoxynucleotides and amino acids. Nuc Acid Res 35:5060–5072
Cheng Y, Prussof WH (1973) Relationship between the inhibition constant (Ki) and the concentration of inhibitor which causes 50 percent inhibition (I50) of an enzymatic reaction. Biochem Pharmacol 22:3099–3108
Hang HC, Yu C, Ten Hagen KG, Tian E, Winans KA, Tabak LA, Bertozzi CR (2004) Small molecule inhibitors of mucin-type O-linked glycosylation from a uridine-based library. Chem Biol 11:337–345
Jiang J, Lazarus MB, Pasquina L, Sliz P, Walker S (2012) A neutral diphosphate mimic crosslinks the active site of human O-GlcNAc transferase. Nat Chem Biol 8:72–77
Magalhaes A, Duarte HO, Reis CA (2021) The role of O-glycosylation in human disease. Mol Aspect Med 79, Article 100964
Olivier T, Haslam A, Prasad V (2021) Anticancer drugs approved by the US food and drug administration from 2009 to 2020. JAMA Netw Open 4:e2138793
Papanicolaou KN, Jung J, Ashok D, Zhang W, Modaressanavi A, Avila A, Foster DB, Zachara NE, O'Rourke B (2023) Inhibiting O-GlcNAcylation impacts p38 and Erk1/2 signalling and perturbs cardiomyocyte hypertrophy. J Biol Chem 299, Article 102907
Pignatti F, Wilking U, Postmus D, Wilking N, Delgado J, Bergh J (2022) The value of anticancer drugs—a regulatory view. Nat Rev Clin Oncol 19:207–215
Prasher P, Pooja P (2014) Singh, Lead modification: amino acid appended indoles as highly effective 5-LOX inhibitors. Bioorg Med Chem 22:1642–1648
Prasher P, Sharma M (2023) Potential strategies for the management of adenocarcinoma: a perspective. Future Med Chem 15:123–127
Qi J, Wang R, Zeng Y, Yu W, Gu Y (2017) New ELISA-based method for the detection of O-GlcNAc transferase activity in vitro. Prep Biochem Biotechnol 47:699–702
Schimpl M, Zheng X, Borodkin VS, Blair DE, Ferenbach AT, Schüttelkopf AW, Navratilova I, Aristotelous T, Albarbarawi O, Robinson DA, Macnaughtan MA, van Aalten DM (2012) O-GlcNAc transferase invokes nucleotide sugar pyrophosphate participation in catalysis. Nat Chem Biol 8:969–974
Sharma M, Prasher P (2022) Selected heterocycles for the development of pyrophosphate mimics. Future Med Chem 14:1219–1222
Siegel RL, Miller KD, Wagle NS, Jemal A (2023) Cancer statistics. CA Cancer J Clin 73:17–48
Singh P, Prasher P, Dhillon P, Bhatti R (2015) Indole based peptidomimetics as anti-inflammatory and anti-hyperalgesic agents: dual inhibition of 5-LOX and COX-2 enzymes. Eur J Med Chem 97:104–123
Singh P, Kaur J, Kaur H, Kaur A, Bhatti R (2018) Synergy of physico-chemical and biological experiments for developing a cyclooxygenase-2 inhibitor. Sci Rep 8, Article 10005
Thomas D, Rathinavel AK, Radhakrishnan P (2021)Altered glycosylation in cancer: a promising target for biomarkers and therapeutics. Biochim Biophys Acta Rev Cancer 1875, Article 188464
Vaghefi MM, Bernacki RJ, Hennen WJ, Robins RK (1987) Synthesis of certain nucleoside methylenediphosphonate sugars as potential inhibitors of glycosyltransferases. J Med Chem 30:1391–1399
Wang R, Steenshma DH, Takaoka Y, Yun JW, Kajimoto T, Wong C-H (1997) A search for pyrophosphate mimics for the development of substrates and inhibitors of glycosyltransferases. Bioorg Med Chem Lett 5:661–672
Acknowledgements
The authors duly acknowledge the Research Grant from DST-SERB, India for supporting this work. PP Thanks DST-SERB for TARE fellowship (File No. TAR/2021/000129).
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Below is the link to the electronic supplementary material.
Rights and permissions
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Mall, T., Sharma, M. & Prasher, P. Amino acid tethered benzoxazolone as highly potent inhibitors of O-glycosylation. Chem. Pap. (2024). https://doi.org/10.1007/s11696-024-03461-y
Received:
Accepted:
Published:
DOI: https://doi.org/10.1007/s11696-024-03461-y