Engineering of a Reductive Aminase to Enable the Synthesis of a Key Intermediate to a CDK 2/4/6 Inhibitor

Gideon James Grogan; Amelia Gilio; Jeremy Steflik; Michael Burns; Rajesh Kumar; Russell Lewis; Carlos Martinez

doi:10.1021/acscatal.3c01534

Engineering of a Reductive Aminase to Enable the Synthesis of a Key Intermediate to a CDK 2/4/6 Inhibitor

Gideon James Grogan, Amelia Gilio, Jeremy Steflik, Michael Burns, Rajesh Kumar, Russell Lewis, Carlos Martinez

Chemistry

Research output: Contribution to journal › Article › peer-review

Abstract

Biocatalytic reductive amination reactions with reductive aminases (RedAms) are emerging transformations with a high potential value for pharmaceutical synthesis. Here, we report the identification and engineering of a RedAm to catalyze a reductive amination reaction, making a key intermediate in the synthesis of an investigational cyclin-dependent kinase (CDK) inhibitor, using the relatively bulky benzylamine as a nucleophile. The engineered enzyme contains six mutations with respect to the wild-type and displays high productivity at high substrate concentrations (50-fold improved over the wild-type). After the optimized enzyme variant was identified, crystal structures of both the wild-type and mutant enzymes were solved and used to rationalize how structural changes to the RedAm improved its performance under process conditions. Results suggest that mutations affecting both substrate binding and enzyme thermostability contribute to improved enzyme performance. By enabling the multikilogram-scale synthesis of the chiral intermediate, this work highlights the versatility and industrial utility of RedAm-catalyzed reductive amination.

Original language	English
Number of pages	11
Journal	ACS Catalysis
Early online date	18 Jul 2023
DOIs	https://doi.org/10.1021/acscatal.3c01534
Publication status	E-pub ahead of print - 18 Jul 2023

Bibliographical note

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10.1021/acscatal.3c01534

Steflik_Accepted_Manuscript_2023
©2023 American Chemical Society. This is an author-produced version of the published paper. Uploaded in accordance with the University’s Research Publications and Open Access policy.
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Cite this

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title = "Engineering of a Reductive Aminase to Enable the Synthesis of a Key Intermediate to a CDK 2/4/6 Inhibitor",

abstract = "Biocatalytic reductive amination reactions with reductive aminases (RedAms) are emerging transformations with a high potential value for pharmaceutical synthesis. Here, we report the identification and engineering of a RedAm to catalyze a reductive amination reaction, making a key intermediate in the synthesis of an investigational cyclin-dependent kinase (CDK) inhibitor, using the relatively bulky benzylamine as a nucleophile. The engineered enzyme contains six mutations with respect to the wild-type and displays high productivity at high substrate concentrations (50-fold improved over the wild-type). After the optimized enzyme variant was identified, crystal structures of both the wild-type and mutant enzymes were solved and used to rationalize how structural changes to the RedAm improved its performance under process conditions. Results suggest that mutations affecting both substrate binding and enzyme thermostability contribute to improved enzyme performance. By enabling the multikilogram-scale synthesis of the chiral intermediate, this work highlights the versatility and industrial utility of RedAm-catalyzed reductive amination.",

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AU - Grogan, Gideon James

AU - Gilio, Amelia

AU - Steflik, Jeremy

AU - Burns, Michael

AU - Kumar, Rajesh

AU - Lewis, Russell

AU - Martinez, Carlos

PY - 2023/7/18

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AB - Biocatalytic reductive amination reactions with reductive aminases (RedAms) are emerging transformations with a high potential value for pharmaceutical synthesis. Here, we report the identification and engineering of a RedAm to catalyze a reductive amination reaction, making a key intermediate in the synthesis of an investigational cyclin-dependent kinase (CDK) inhibitor, using the relatively bulky benzylamine as a nucleophile. The engineered enzyme contains six mutations with respect to the wild-type and displays high productivity at high substrate concentrations (50-fold improved over the wild-type). After the optimized enzyme variant was identified, crystal structures of both the wild-type and mutant enzymes were solved and used to rationalize how structural changes to the RedAm improved its performance under process conditions. Results suggest that mutations affecting both substrate binding and enzyme thermostability contribute to improved enzyme performance. By enabling the multikilogram-scale synthesis of the chiral intermediate, this work highlights the versatility and industrial utility of RedAm-catalyzed reductive amination.

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