A Novel One-Pot Enzyme Cascade for the Biosynthesis of Cladribine Triphosphate.

Julia Frisch,Tin Maršić,Christoph Loderer

doi:10.3390/biom11030346

Julia Frisch, Tin Maršić + Show 1 more

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https://doi.org/10.3390/biom11030346

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Abstract

Cladribine triphosphate is the active compound of the anti-cancer and multiple sclerosis drug Mavenclad (cladribine). Biosynthesis of such non-natural deoxyribonucleotides is challenging but important in order to study the pharmaceutical modes of action. In this study, we developed a novel one-pot enzyme cascade for the biosynthesis of cladribine triphosphate, starting with the nucleobase 2Cl-adenine and the generic co-substrate phosphoribosyl pyrophosphate. The cascade is comprised of the three enzymes, namely, adenine phosphoribosyltransferase (APT), polyphosphate kinase (PPK), and ribonucleotide reductase (RNR). APT catalyzes the binding of the nucleobase to the ribose moiety, followed by two consecutive phosphorylation reactions by PPK. The formed nucleoside triphosphate is reduced to the final product 2Cl-deoxyadenonsine triphosphate (cladribine triphosphate) by the RNR. The cascade is feasible, showing comparative product concentrations and yields to existing enzyme cascades for nucleotide biosynthesis. While this study is limited to the biosynthesis of cladribine triphosphate, the design of the cascade offers the potential to extend its application to other important deoxyribonucleotides.

Highlights

We report a new biocatalytic approach for the biosynthesis of nonnatural deoxyribonucleotides
The objective of this study is to explore the feasibility of this cascade for the small-scale production of cladribine triphosphate, the active metabolite of the anti-cancer, and multiple sclerosis drug cladribine
Phosphoribosyltransferases are a well-established tool for nucleoside monophosphate biosynthesis and have been applied in numerous studies [12,13,14,15,16,17]

Summary

Introduction

Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations. On 29 March 2019 the pharmaceutical Mavenclad (cladribine) was approved by the. U.S Food and Drug Administration (FDA) as treatment against relapsing forms of multiple sclerosis. Cladribine was developed in the 1980s as a treatment for different forms of leukemia [1], and only 30 years later, its potential against multiple sclerosis was realized and studied in detail [2,3]. Its mode of action revolves around its cytotoxicity by induction of DNA strand breaks [3,4]. The detailed modes of action are subject of current research [4,5]

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