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. 2023 Oct 11;145(40):22041-22046.
doi: 10.1021/jacs.3c07010. Epub 2023 Oct 2.

Biocatalysis in Drug Design: Engineered Reductive Aminases (RedAms) Are Used to Access Chiral Building Blocks with Multiple Stereocenters

Arnau Rué Casamajo  1 , Yuqi Yu  1 , Christian Schnepel  2 , Charlotte Morrill  1 , Rhys Barker  1 , Colin W Levy  1 , James Finnigan  3 , Victor Spelling  4 , Kristina Westerlund  5 , Mark Petchey  6 , Robert J Sheppard  5 , Richard J Lewis  7 , Francesco Falcioni  8 , Martin A Hayes  6 , Nicholas J Turner  1
Affiliations

Biocatalysis in Drug Design: Engineered Reductive Aminases (RedAms) Are Used to Access Chiral Building Blocks with Multiple Stereocenters

Arnau Rué Casamajo et al. J Am Chem Soc. .

Abstract

Novel building blocks are in constant demand during the search for innovative bioactive small molecule therapeutics by enabling the construction of structure-activity-property-toxicology relationships. Complex chiral molecules containing multiple stereocenters are an important component in compound library expansion but can be difficult to access by traditional organic synthesis. Herein, we report a biocatalytic process to access a specific diastereomer of a chiral amine building block used in drug discovery. A reductive aminase (RedAm) was engineered following a structure-guided mutagenesis strategy to produce the desired isomer. The engineered RedAm (IR-09 W204R) was able to generate the (S,S,S)-isomer 3 in 45% conversion and 95% ee from the racemic ketone 2. Subsequent palladium-catalyzed deallylation of 3 yielded the target primary amine 4 in a 73% yield. This engineered biocatalyst was used at preparative scale and represents a potential starting point for further engineering and process development.

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Conflict of interest statement

The authors declare no competing financial interest.

Figures

Figure 1
Figure 1
Combined biocatalytic resolution and reductive amination of racemic ketone 2 with allylamine 1 forming (S,S,S)-3, which is a key intermediate for the desired primary amine (S,S,S)-4.
Figure 2
Figure 2
(a) The four possible reductive amination products (S,S,S)-3, (R,R,R)-3, (S,S,R)-3, and (R,R,S)-3 derived from ketone (±)-2; (b) SFC chromatogram of the stereoisomers.
Figure 3
Figure 3
(a) Crystal structure of IRED-09 (purple) in complex with NADPH (pink) and N-cyclopropylcyclohexanamine; there is only one molecule in the asymmetric unit. (b) IR-09 biological dimer. (c) Active site of IR-09 with the imine intermediate of 3 modeled into the active site showing distances (Å) from C4 of the nicotinamide ring of NAPDH and (B) W204 to the electrophilic carbon of the ligand. (d) The view is rotated 180 deg to observe the active site from the opposite perspective and show distances from (B) M233 and (B) Q234 to the ligand.
Figure 4
Figure 4
(a) SFC chromatograms comparing the isomer production of IR-09 WT, the best SDM variant, and the best SSM variant (peak at rt = 4.4. min corresponds to the alcohol). (b) Comparison between WT and the best variants for the production of (S,S,S)-3. Conversion to (S,S,S)-3 (%) = (S,S,S) yield × conversion.
See this image and copyright information in PMC

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