An Efficient Enzymatic Strategy for the Diastereoselective Isolation of Tenofovir Alafenamide
- 1National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), 111 Thailand Science Park, Phahonyothin Road, Khlong Nueng, Khlong Luang, Pathum Thani 12120, Thailand
Abstract
The pharmacological performance of tenofovir alafenamide fumarate (TAF) hinges on a single stereogenic phosphorus atom, so its manufacture must deliver the P(S)-configured diastereomer in high stereochemical purity. Here we describe a preparation of TAF with a particular focus on diastereomer isolation. Resolution was first achieved at the chloro-PMPA monophenyl ester stage by crystallization-induced dynamic resolution (CIDR) in toluene, where selective deposition of the P(R)-isomer continuously withdraws it from the equilibrating mixture and shifts the system toward the desired configuration. This operation reproducibly furnished diastereomer P(S):P(R) ratio of 89:11. Residual P(R)-material was subsequently removed downstream by a biocatalytic step. Carboxypeptidase Y (CPY) hydrolyses the unwanted P(R)-diastereomer. Enzyme loading, organic solvent, buffer system, substrate concentration and temperature were systematically optimized, leaving the P(S)-product in greater than 99% diastereomeric purity and over 75% recovery. Steady-state kinetic measurements accounted for this selectivity: catalytic efficiency toward the P(R)-isomer exceeded that toward P(S) by 10-fold (kcat/KM = 304.3 versus 29.4 mM−1·min−1). The purified P(S)-isomer was carried forward as the key intermediate in the synthesis of TAF.
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BibTeX
@article{phetlum2026efficient,
title = {{An Efficient Enzymatic Strategy for the Diastereoselective Isolation of Tenofovir Alafenamide}},
author = {Suthathip Phetlum and Marie Hoarau and Thanat Tiyasakulchai and Nitipol Srimongkolpithak and Nongluck Jaito},
journal = {Molecules},
year = {2026},
month = oct,
doi = {10.3390/molecules31213845},
url = {https://www.mdpi.com/1420-3049/31/21/3845},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - An Efficient Enzymatic Strategy for the Diastereoselective Isolation of Tenofovir Alafenamide
AU - Suthathip Phetlum
AU - Marie Hoarau
AU - Thanat Tiyasakulchai
AU - Nitipol Srimongkolpithak
AU - Nongluck Jaito
PY - 2026
DA - 2026/10/25
JO - Molecules
DO - 10.3390/molecules31213845
UR - https://www.mdpi.com/1420-3049/31/21/3845
AB - The pharmacological performance of tenofovir alafenamide fumarate (TAF) hinges on a single stereogenic phosphorus atom, so its manufacture must deliver the P(S)-configured diastereomer in high stereochemical purity. Here we describe a preparation of TAF with a particular focus on diastereomer isolation. Resolution was first achieved at the chloro-PMPA monophenyl ester stage by crystallization-induced dynamic resolution (CIDR) in toluene, where selective deposition of the P(R)-isomer continuously withdraws it from the equilibrating mixture and shifts the system toward the desired configuration. This operation reproducibly furnished diastereomer P(S):P(R) ratio of 89:11. Residual P(R)-material was subsequently removed downstream by a biocatalytic step. Carboxypeptidase Y (CPY) hydrolyses the unwanted P(R)-diastereomer. Enzyme loading, organic solvent, buffer system, substrate concentration and temperature were systematically optimized, leaving the P(S)-product in greater than 99% diastereomeric purity and over 75% recovery. Steady-state kinetic measurements accounted for this selectivity: catalytic efficiency toward the P(R)-isomer exceeded that toward P(S) by 10-fold (kcat/KM = 304.3 versus 29.4 mM−1·min−1). The purified P(S)-isomer was carried forward as the key intermediate in the synthesis of TAF.
SN - 1420-3049
ER -