176655-55-3Relevant articles and documents
Recombinant expression and characterization of novel P450s from Actinosynnema mirum
Schmitz, Lisa Marie,Hageneier, Felix,Rosenthal, Katrin,Busche, Tobias,Brandt, David,Kalinowski, J?rn,Lütz, Stephan
, (2021)
Cytochrome P450 monooxygenases (P450s) are the major contributor in the metabolism of xenobiotics, including therapeutic agents. Thus, P450s find broad application in the pharmaceutical industry to synthesize metabolites of new active pharmaceutical ingredients in order to evaluate toxicity and pharmacokinetics. As an alternative to human hepatic P450s, microbial P450s offer several advantages, such as an easier and more efficient heterologous expression as well as higher stability under process conditions. Recently, the wild-type strain Actinosynnema mirum has been reported to catalyze hydroxylation reactions with high activity on a broad range of substrates. In this study, one of these substrates, ritonavir, was used to analyze the transcriptional response of the wild-type strain. Analysis of the differential gene expression pattern allowed the assignment of genes potentially responsible for ritonavir conversion. Heterologous expression of these candidates and activity testing led to the identification of a novel P450 that efficiently converts ritonavir resembling the activity of the human CYP3A4.
The effect of ritonavir on human CYP2B6 catalytic activity: Heme modification contributes to the mechanism-based inactivation of CYP2B6 and CYP3A4 by ritonavir
Lin, Hsia-Lien,D'Agostino, Jaime,Kenaan, Cesar,Calinski, Diane,Hollenberg, Paul F.
, p. 1813 - 1824 (2013)
The mechanism-based inactivation of human CYP2B6 by ritonavir (RTV) in a reconstituted system was investigated. The inactivation is time, concentration, and NADPH dependent and exhibits a Kl of 0.9 μM, a k inact of 0.05 min-1, and a partition ratio of approximately 3. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis showed that the protonated molecular ion of RTV exhibits an m/z at 721 and its two major metabolites are an oxidation product with MH+ at m/z 737 and a deacylated product with MH+at m/z 580. Inactivation of CYP2B6 by incubation with 10 μM RTV for 10 min resulted in an approximately 50% loss of catalytic activity and native heme, but no modification of the apoprotein was observed. RTV was found to be a potent mixed-type reversible inhibitor (Ki = 0.33 μM) and a type II ligand (spectral dissociation constant-Ks = 0.85 mM) of CYP2B6. Although previous studies have demonstrated that RTV is a potent mechanism-based inactivator of CYP3A4, the molecular mechanism responsible for the inactivation has not been determined. Here, we provide evidence that RTV inactivation of CYP3A4 is due to heme destruction with the formation of a heme-protein adduct. Similar to CYP2B6, there is no significant modification of the apoprotein. Furthermore, LC-MS/MS analysis revealed that both CYP3A4 and human liver microsomes form an RTV-glutathione conjugate having a MH+ at m/z 858 during metabolism of RTV, suggesting the formation of an isocyanate intermediate leading to formation of the conjugate. Copyright
Explanation through density functional theory of the unanticipated loss of CO2 and differences in mass fragmentation profiles of ritonavir and its rCYP3A4-mediated metabolites
Jhajra, Shalu,Handa, Tarun,Bhatia, Sonam,Bharatam,Singh, Saranjit
, p. 452 - 467 (2014)
In the present study, the metabolism of ritonavir was explored in the presence of rCYP3A4 using a well-established strategy involving liquid chromatography-mass spectrometry (LC-MS) tools. A total of six metabolites were formed, of which two were new, not reported earlier as CYP3A4-mediated metabolites. During LC-MS studies, ritonavir was found to fragment through six principal pathways, many of which involved neutral loss of CO2, as indicated through 44-Da difference between masses of the precursors and the product ions. This was unusual as the drug and the precursors were devoid of a terminal carboxylic acid group. Apart from the neutral loss of CO2, marked differences were also observed among the fragmentation pathways of the drug and its metabolites having intact N-methyl moiety as compared to those lacking N-methyl moiety. These unusual fragmentation behaviours were successfully explained through energy distribution profiles by application of the density functional theory.
Preparation method of ritonavir
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, (2021/10/13)
The invention provides a preparation method shown I, N - [(2S, 3S, 5R) -3 - hydroxyl -5 - [(2S) -3 - methyl -2 - [[methyl - [(2 - isopropyl -1, 3 - thiazole -4 - yl) methyl] carbamoyl] amino] -1, 6 - diphenyl - hexyl -2 -] carbamic acid 5 - thiazolyl methyl ester. The invention provides a new method for preparing ritonavir.
Preparation method of ritonavir
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, (2018/07/30)
The invention discloses a preparation method of ritonavir. According to the preparation method, N-[N-Methyl-N-((2-isopropyl-4-thiazolyl)methyl)aminocarbonyl]-L-valine is taken as a raw material, and is reacted with thionyl chloride so as to obtain an intermediate I; the intermediate I is reacted with ((2S, 3S, 5S)-5-amino-3-hydroxy-1.6-diphenylhexane-2-yl) tert-butyl carbamate so as to obtain intermediate II; the intermediate II is subjected to hydrolysis to remove tertbutyloxycarbonyl so as to obtain an intermediate III; the intermediate III is reacted with ((5-Thiazolyl)methyl)-(4-nitrophenyl)carbonate so as to obtain finished product ritonavir. The preparation method is short in synthesis route; the raw materials are simple and are easily available; the reaction sequence is reasonable;using of expensive palladium catalyst and condensing agents such as DCC, EDC, HATU, DEPBT, and toxic triphosgene is avoided; the preparation method is green, is friendly to the environment; reaction conditions are mild; side product is less; synthesis yield is high; the purity of obtain ritonavir is high; production cost is low; and the preparation method is suitable for industrialized production.
