Speaker
Description
Submission Type: Poster Presentation
Submitted by: Hae Chan Jeong (Chonnam National University, KR)
Track: New Tools for Studying Drug Metabolism
Biocatalytic synthesis of a novel atorvastatin derivative as an anti-hyperlipidemic drug candidate using sequential reaction of P450 and tyrosinase
Hae Chan Jeong1, Yu-Jin Lee1, Gun Su Cha2, Fikri A. R. Hardiyanti Oktavia1, Chan Mi Park2, Chul-Ho Yun1,3
1School of Biological Sciences and Biotechnology, Graduate School, Chonnam National University, 77 Yongbongro, Gwangju 61186, Republic of Korea
2Institute of Natural BIO Industry of Namwon, 43 Simyogil, Namwonsi, Jeonbukdo 55801, Republic of Korea
3School of Biological Sciences and Technology, Chonnam National University, 77 Yongbongro, Gwangju 61186, Republic of Korea
E-mail: hcjeong450@gmail.com
[Background]
Atorvastatin effectively lowers total LDL-cholesterol levels by competitively inhibiting HMG-CoA reductase. Its active metabolite, 2-hydroxy and 4-hydroxy atorvastatin (4-OH ATO), produced by cytochrome P450s, also exhibit inhibitory effects on HMG-CoA reductase. Similar to atorvastatin, certain drugs undergo aromatic hydroxylation to produce active metabolites having pharmacological potencies that are equivalent to or significantly greater than those of their parent compounds.
Inspired by in vivo metabolism, derivatives of parent drug scaffolds are attractive candidates for novel drug development, offering clinically validated structures that retain pharmacological activity with potentially lower risk profiles. In particular, aromatic hydroxylation of a phenolic moiety is an important bioactivation process, producing catechol metabolites from Phase Ⅰ-derived phenolic intermediates. Chemoenzymatic or dual-enzymatic systems provide regio- and stereoselective synthesis under mild aqueous conditions and enable efficient production of complex derivatives.
[Methods]
4-OH ATO was produced from atorvastatin using the heme domain of CYP102A1 mutant. Then, 4-OH ATO was subsequently converted to 3,4-dihydroxy atorvastatin (3,4-diOH ATO) by tyrosinase derived from Priestia megaterium GJ804. The products were isolated by HPLC and identified by LC-MS and NMR analysis. The inhibitory activities of atorvastatin and its derivatives against HMG-CoA reductase were evaluated by in vitro inhibition assay.
[Results]
The H1120 mutant catalyzes the para-hydroxylation of atorvastatin utilizing H2O2 as a co-substrate, bypassing the need for complex reductase systems and NADPH. This H1120-based system achieved the highest reported productivity for 4-OH ATO (121 μM) to date. The subsequent conversion of 4-OH ATO to 3,4-diOH ATO by GJ804 Ty was demonstrated for the first time. Despite the disparate optimal conditions for the two enzymes, a stepwise one-pot cascade system for the synthesis of new atorvastatin derivatives was successfully established. It was also confirmed that 4-OH ATO is converted to 3,4-diOH ATO by human liver microsomes (HLMs). The novel derivative, 3,4-diOH ATO, retains HMG-CoA reductase inhibitory activity comparable to both atorvastatin and 4-OH ATO.
[Conclusions]
We report the biocatalytic synthesis of a new atorvastatin derivative. Using an engineered heme domain of P450 BM3 (CYP102A1), atorvastatin was converted to 4-OH ATO, which was subsequently transformed into 3,4-diOH ATO using a bacterial tyrosinase. Notably, 3,4-diOH ATO exhibited HMG-CoA reductase inhibitory activity comparable to both atorvastatin and 4-OH ATO. This study demonstrates the potential of biocatalysis for the selective synthesis of active atorvastatin derivatives as a platform for discovering novel drug candidates.
[Acknowledgements]
This research was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Science, ICT & Future Planning (CHY: No. RS-2021-NR059615 and RS-2024-00440681) and the Ministry of Education (HCJ: RS-2025-25426590)
