Discovery of Novel Reductive Elimination Pathway for 10-Hydroxywarfarin
Coumadin (R/S-warfarin) anticoagulant therapy is highly efficacious in preventing the formation of blood clots; however, significant inter-individual variations in response risks over or under dosing resulting in adverse bleeding events or ineffective therapy, respectively. Levels of pharmacological...
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| Format: | Article |
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Frontiers Media S.A.
2022-01-01
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| Series: | Frontiers in Pharmacology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fphar.2021.805133/full |
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| author | Dakota L. Pouncey Dustyn A. Barnette Riley W. Sinnott Sarah J. Phillips Noah R. Flynn Howard P. Hendrickson Howard P. Hendrickson S. Joshua Swamidass Grover P. Miller |
| author_facet | Dakota L. Pouncey Dustyn A. Barnette Riley W. Sinnott Sarah J. Phillips Noah R. Flynn Howard P. Hendrickson Howard P. Hendrickson S. Joshua Swamidass Grover P. Miller |
| author_sort | Dakota L. Pouncey |
| collection | DOAJ |
| description | Coumadin (R/S-warfarin) anticoagulant therapy is highly efficacious in preventing the formation of blood clots; however, significant inter-individual variations in response risks over or under dosing resulting in adverse bleeding events or ineffective therapy, respectively. Levels of pharmacologically active forms of the drug and metabolites depend on a diversity of metabolic pathways. Cytochromes P450 play a major role in oxidizing R- and S-warfarin to 6-, 7-, 8-, 10-, and 4′-hydroxywarfarin, and warfarin alcohols form through a minor metabolic pathway involving reduction at the C11 position. We hypothesized that due to structural similarities with warfarin, hydroxywarfarins undergo reduction, possibly impacting their pharmacological activity and elimination. We modeled reduction reactions and carried out experimental steady-state reactions with human liver cytosol for conversion of rac-6-, 7-, 8-, 4′-hydroxywarfarin and 10-hydroxywarfarin isomers to the corresponding alcohols. The modeling correctly predicted the more efficient reduction of 10-hydroxywarfarin over warfarin but not the order of the remaining hydroxywarfarins. Experimental studies did not indicate any clear trends in the reduction for rac-hydroxywarfarins or 10-hydroxywarfarin into alcohol 1 and 2. The collective findings indicated the location of the hydroxyl group significantly impacted reduction selectivity among the hydroxywarfarins, as well as the specificity for the resulting metabolites. Based on studies with R- and S-7-hydroxywarfarin, we predicted that all hydroxywarfarin reductions are enantioselective toward R substrates and enantiospecific for S alcohol metabolites. CBR1 and to a lesser extent AKR1C3 reductases are responsible for those reactions. Due to the inefficiency of reactions, only reduction of 10-hydroxywarfarin is likely to be important in clearance of the metabolite. This pathway for 10-hydroxywarfarin may have clinical relevance as well given its anticoagulant activity and capacity to inhibit S-warfarin metabolism. |
| first_indexed | 2024-12-18T05:25:05Z |
| format | Article |
| id | doaj.art-a8dec47d9bf6442485f60c67345fb8f1 |
| institution | Directory Open Access Journal |
| issn | 1663-9812 |
| language | English |
| last_indexed | 2024-12-18T05:25:05Z |
| publishDate | 2022-01-01 |
| publisher | Frontiers Media S.A. |
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| series | Frontiers in Pharmacology |
| spelling | doaj.art-a8dec47d9bf6442485f60c67345fb8f12022-12-21T21:19:33ZengFrontiers Media S.A.Frontiers in Pharmacology1663-98122022-01-011210.3389/fphar.2021.805133805133Discovery of Novel Reductive Elimination Pathway for 10-HydroxywarfarinDakota L. Pouncey0Dustyn A. Barnette1Riley W. Sinnott2Sarah J. Phillips3Noah R. Flynn4Howard P. Hendrickson5Howard P. Hendrickson6S. Joshua Swamidass7Grover P. Miller8Department of Biochemistry and Molecular Biology, College of Medicine, University of Arkansas for Medical Sciences, Little Rock, AR, United StatesDepartment of Biochemistry and Molecular Biology, College of Medicine, University of Arkansas for Medical Sciences, Little Rock, AR, United StatesDepartment of Biochemistry and Molecular Biology, College of Medicine, University of Arkansas for Medical Sciences, Little Rock, AR, United StatesDepartment of Pharmaceutical Sciences, College of Pharmacy, University of Arkansas for Medical Sciences, Little Rock, AR, United StatesDepartment of Pathology and Immunology, Washington University School of Medicine, St. Louis, MO, United StatesDepartment of Pharmaceutical Sciences, College of Pharmacy, University of Arkansas for Medical Sciences, Little Rock, AR, United StatesDepartment of Pharmaceutical Social and Administrative Sciences, McWhorter School of Pharmacy, Samford University, Birmingham, AL, United StatesDepartment of Pathology and Immunology, Washington University School of Medicine, St. Louis, MO, United StatesDepartment of Biochemistry and Molecular Biology, College of Medicine, University of Arkansas for Medical Sciences, Little Rock, AR, United StatesCoumadin (R/S-warfarin) anticoagulant therapy is highly efficacious in preventing the formation of blood clots; however, significant inter-individual variations in response risks over or under dosing resulting in adverse bleeding events or ineffective therapy, respectively. Levels of pharmacologically active forms of the drug and metabolites depend on a diversity of metabolic pathways. Cytochromes P450 play a major role in oxidizing R- and S-warfarin to 6-, 7-, 8-, 10-, and 4′-hydroxywarfarin, and warfarin alcohols form through a minor metabolic pathway involving reduction at the C11 position. We hypothesized that due to structural similarities with warfarin, hydroxywarfarins undergo reduction, possibly impacting their pharmacological activity and elimination. We modeled reduction reactions and carried out experimental steady-state reactions with human liver cytosol for conversion of rac-6-, 7-, 8-, 4′-hydroxywarfarin and 10-hydroxywarfarin isomers to the corresponding alcohols. The modeling correctly predicted the more efficient reduction of 10-hydroxywarfarin over warfarin but not the order of the remaining hydroxywarfarins. Experimental studies did not indicate any clear trends in the reduction for rac-hydroxywarfarins or 10-hydroxywarfarin into alcohol 1 and 2. The collective findings indicated the location of the hydroxyl group significantly impacted reduction selectivity among the hydroxywarfarins, as well as the specificity for the resulting metabolites. Based on studies with R- and S-7-hydroxywarfarin, we predicted that all hydroxywarfarin reductions are enantioselective toward R substrates and enantiospecific for S alcohol metabolites. CBR1 and to a lesser extent AKR1C3 reductases are responsible for those reactions. Due to the inefficiency of reactions, only reduction of 10-hydroxywarfarin is likely to be important in clearance of the metabolite. This pathway for 10-hydroxywarfarin may have clinical relevance as well given its anticoagulant activity and capacity to inhibit S-warfarin metabolism.https://www.frontiersin.org/articles/10.3389/fphar.2021.805133/fullanticoagulationwarfarinhydroxywarfarinmetabolismmodelingreduction |
| spellingShingle | Dakota L. Pouncey Dustyn A. Barnette Riley W. Sinnott Sarah J. Phillips Noah R. Flynn Howard P. Hendrickson Howard P. Hendrickson S. Joshua Swamidass Grover P. Miller Discovery of Novel Reductive Elimination Pathway for 10-Hydroxywarfarin Frontiers in Pharmacology anticoagulation warfarin hydroxywarfarin metabolism modeling reduction |
| title | Discovery of Novel Reductive Elimination Pathway for 10-Hydroxywarfarin |
| title_full | Discovery of Novel Reductive Elimination Pathway for 10-Hydroxywarfarin |
| title_fullStr | Discovery of Novel Reductive Elimination Pathway for 10-Hydroxywarfarin |
| title_full_unstemmed | Discovery of Novel Reductive Elimination Pathway for 10-Hydroxywarfarin |
| title_short | Discovery of Novel Reductive Elimination Pathway for 10-Hydroxywarfarin |
| title_sort | discovery of novel reductive elimination pathway for 10 hydroxywarfarin |
| topic | anticoagulation warfarin hydroxywarfarin metabolism modeling reduction |
| url | https://www.frontiersin.org/articles/10.3389/fphar.2021.805133/full |
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