Atypical Kinetics and Albumin Effect of Glucuronidation of 5-n-Butyl-4-{4-[2-(1H-tetrazole-5- yl)-1H-pyrrol-1-yl]phenylmethyl}-2,4-dihydro-2-(2,6- dichlorophenyl)-3H-1,2,4-triazol-3-one, a Novel Nonpeptide Angiotensin Type 1 Receptor Antagonist, in Liver Microsomes and UDP-Glucuronosyl-transferase
Ib is a new nonpeptide AT1 receptor antagonist, which plays an active role in cardiovascular protection. Ib monoglucuronide has been identified as its main metabolite. A detailed study of Ib glucuronidation is important for predicting potential DDI. Besides, the elucidation of the “BSA effect” in Ib...
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MDPI AG
2018-03-01
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| Series: | Molecules |
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| Online Access: | http://www.mdpi.com/1420-3049/23/3/688 |
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| author | Ying Peng Xueyuan Zhang Yinci Zhu Hui Wu Shiyin Gu Qingqing Chang Yi Zhou Guangji Wang Jianguo Sun |
| author_facet | Ying Peng Xueyuan Zhang Yinci Zhu Hui Wu Shiyin Gu Qingqing Chang Yi Zhou Guangji Wang Jianguo Sun |
| author_sort | Ying Peng |
| collection | DOAJ |
| description | Ib is a new nonpeptide AT1 receptor antagonist, which plays an active role in cardiovascular protection. Ib monoglucuronide has been identified as its main metabolite. A detailed study of Ib glucuronidation is important for predicting potential DDI. Besides, the elucidation of the “BSA effect” in Ib glucuronidation would make obtained kinetic parameters more predictive in IVIVE. “BSA effect” means that there is a significant change in in vitro kinetic parameters when generated from incubations performed in the presence of bovine serum albumin (BSA). Five UGTs (UGT1A3, UGT2B4, UGT2B7, UGT1A9 and UGT1A8) were identified that produced abundant Ib monoglucuronide, especially UGT1A3. We investigated Ib glucuronidation in liver microsomes from different species (rat, dog, human) and in five identified major human UGTs. Ib glucuronidation in liver microsomes and recombinant human UGTs all showed substrate inhibition kinetics. DLM showed the strongest affinity and activity, HLM showed the lowest affinity, and RLM showed the weakest activity. The addition of BSA did not alter the enzyme kinetics, but significantly altered enzyme kinetic parameters resulting in a reduction in Km value and an increase in CLint value. However, high concentrations of BSA could significantly attenuate this positive effect on enzyme affinity and activity, and the effect of BSA on the Vmax of Ib glucuronidation was opposite in different enzyme sources. In conclusion, this study demonstrated the substrate inhibition kinetics of Ib glucuronidation in the liver metabolism and the effect of BSA on its kinetic parameters, in order to provide more accurate in vitro data for in vivo prediction. |
| first_indexed | 2024-04-12T13:48:18Z |
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| language | English |
| last_indexed | 2024-04-12T13:48:18Z |
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| spelling | doaj.art-3029b93a23314e9aa77e3ccabd2526d62022-12-22T03:30:36ZengMDPI AGMolecules1420-30492018-03-0123368810.3390/molecules23030688molecules23030688Atypical Kinetics and Albumin Effect of Glucuronidation of 5-n-Butyl-4-{4-[2-(1H-tetrazole-5- yl)-1H-pyrrol-1-yl]phenylmethyl}-2,4-dihydro-2-(2,6- dichlorophenyl)-3H-1,2,4-triazol-3-one, a Novel Nonpeptide Angiotensin Type 1 Receptor Antagonist, in Liver Microsomes and UDP-Glucuronosyl-transferaseYing Peng0Xueyuan Zhang1Yinci Zhu2Hui Wu3Shiyin Gu4Qingqing Chang5Yi Zhou6Guangji Wang7Jianguo Sun8Key Lab of Drug Metabolism and Pharmacokinetics, State Key Laboratory of Natural Medicines, Pharmaceutical University China, Nanjing 210009, Jiangsu, ChinaKey Lab of Drug Metabolism and Pharmacokinetics, State Key Laboratory of Natural Medicines, Pharmaceutical University China, Nanjing 210009, Jiangsu, ChinaKey Lab of Drug Metabolism and Pharmacokinetics, State Key Laboratory of Natural Medicines, Pharmaceutical University China, Nanjing 210009, Jiangsu, ChinaKey Lab of Drug Metabolism and Pharmacokinetics, State Key Laboratory of Natural Medicines, Pharmaceutical University China, Nanjing 210009, Jiangsu, ChinaKey Lab of Drug Metabolism and Pharmacokinetics, State Key Laboratory of Natural Medicines, Pharmaceutical University China, Nanjing 210009, Jiangsu, ChinaKey Lab of Drug Metabolism and Pharmacokinetics, State Key Laboratory of Natural Medicines, Pharmaceutical University China, Nanjing 210009, Jiangsu, ChinaKey Lab of Drug Metabolism and Pharmacokinetics, State Key Laboratory of Natural Medicines, Pharmaceutical University China, Nanjing 210009, Jiangsu, ChinaKey Lab of Drug Metabolism and Pharmacokinetics, State Key Laboratory of Natural Medicines, Pharmaceutical University China, Nanjing 210009, Jiangsu, ChinaKey Lab of Drug Metabolism and Pharmacokinetics, State Key Laboratory of Natural Medicines, Pharmaceutical University China, Nanjing 210009, Jiangsu, ChinaIb is a new nonpeptide AT1 receptor antagonist, which plays an active role in cardiovascular protection. Ib monoglucuronide has been identified as its main metabolite. A detailed study of Ib glucuronidation is important for predicting potential DDI. Besides, the elucidation of the “BSA effect” in Ib glucuronidation would make obtained kinetic parameters more predictive in IVIVE. “BSA effect” means that there is a significant change in in vitro kinetic parameters when generated from incubations performed in the presence of bovine serum albumin (BSA). Five UGTs (UGT1A3, UGT2B4, UGT2B7, UGT1A9 and UGT1A8) were identified that produced abundant Ib monoglucuronide, especially UGT1A3. We investigated Ib glucuronidation in liver microsomes from different species (rat, dog, human) and in five identified major human UGTs. Ib glucuronidation in liver microsomes and recombinant human UGTs all showed substrate inhibition kinetics. DLM showed the strongest affinity and activity, HLM showed the lowest affinity, and RLM showed the weakest activity. The addition of BSA did not alter the enzyme kinetics, but significantly altered enzyme kinetic parameters resulting in a reduction in Km value and an increase in CLint value. However, high concentrations of BSA could significantly attenuate this positive effect on enzyme affinity and activity, and the effect of BSA on the Vmax of Ib glucuronidation was opposite in different enzyme sources. In conclusion, this study demonstrated the substrate inhibition kinetics of Ib glucuronidation in the liver metabolism and the effect of BSA on its kinetic parameters, in order to provide more accurate in vitro data for in vivo prediction.http://www.mdpi.com/1420-3049/23/3/688Ib glucuronidationenzyme kineticsBSA effectliver microsomesUGTs |
| spellingShingle | Ying Peng Xueyuan Zhang Yinci Zhu Hui Wu Shiyin Gu Qingqing Chang Yi Zhou Guangji Wang Jianguo Sun Atypical Kinetics and Albumin Effect of Glucuronidation of 5-n-Butyl-4-{4-[2-(1H-tetrazole-5- yl)-1H-pyrrol-1-yl]phenylmethyl}-2,4-dihydro-2-(2,6- dichlorophenyl)-3H-1,2,4-triazol-3-one, a Novel Nonpeptide Angiotensin Type 1 Receptor Antagonist, in Liver Microsomes and UDP-Glucuronosyl-transferase Molecules Ib glucuronidation enzyme kinetics BSA effect liver microsomes UGTs |
| title | Atypical Kinetics and Albumin Effect of Glucuronidation of 5-n-Butyl-4-{4-[2-(1H-tetrazole-5- yl)-1H-pyrrol-1-yl]phenylmethyl}-2,4-dihydro-2-(2,6- dichlorophenyl)-3H-1,2,4-triazol-3-one, a Novel Nonpeptide Angiotensin Type 1 Receptor Antagonist, in Liver Microsomes and UDP-Glucuronosyl-transferase |
| title_full | Atypical Kinetics and Albumin Effect of Glucuronidation of 5-n-Butyl-4-{4-[2-(1H-tetrazole-5- yl)-1H-pyrrol-1-yl]phenylmethyl}-2,4-dihydro-2-(2,6- dichlorophenyl)-3H-1,2,4-triazol-3-one, a Novel Nonpeptide Angiotensin Type 1 Receptor Antagonist, in Liver Microsomes and UDP-Glucuronosyl-transferase |
| title_fullStr | Atypical Kinetics and Albumin Effect of Glucuronidation of 5-n-Butyl-4-{4-[2-(1H-tetrazole-5- yl)-1H-pyrrol-1-yl]phenylmethyl}-2,4-dihydro-2-(2,6- dichlorophenyl)-3H-1,2,4-triazol-3-one, a Novel Nonpeptide Angiotensin Type 1 Receptor Antagonist, in Liver Microsomes and UDP-Glucuronosyl-transferase |
| title_full_unstemmed | Atypical Kinetics and Albumin Effect of Glucuronidation of 5-n-Butyl-4-{4-[2-(1H-tetrazole-5- yl)-1H-pyrrol-1-yl]phenylmethyl}-2,4-dihydro-2-(2,6- dichlorophenyl)-3H-1,2,4-triazol-3-one, a Novel Nonpeptide Angiotensin Type 1 Receptor Antagonist, in Liver Microsomes and UDP-Glucuronosyl-transferase |
| title_short | Atypical Kinetics and Albumin Effect of Glucuronidation of 5-n-Butyl-4-{4-[2-(1H-tetrazole-5- yl)-1H-pyrrol-1-yl]phenylmethyl}-2,4-dihydro-2-(2,6- dichlorophenyl)-3H-1,2,4-triazol-3-one, a Novel Nonpeptide Angiotensin Type 1 Receptor Antagonist, in Liver Microsomes and UDP-Glucuronosyl-transferase |
| title_sort | atypical kinetics and albumin effect of glucuronidation of 5 n butyl 4 4 2 1h tetrazole 5 yl 1h pyrrol 1 yl phenylmethyl 2 4 dihydro 2 2 6 dichlorophenyl 3h 1 2 4 triazol 3 one a novel nonpeptide angiotensin type 1 receptor antagonist in liver microsomes and udp glucuronosyl transferase |
| topic | Ib glucuronidation enzyme kinetics BSA effect liver microsomes UGTs |
| url | http://www.mdpi.com/1420-3049/23/3/688 |
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