Cloning and in vivo metabolizing activity study of CYP3A4 on amiodarone drug residues: A possible probiotic and therapeutic option - 30/05/20
Bienvenue sur EM-consulte, la référence des professionnels de santé.
L’accès au texte intégral de cet article nécessite un abonnement.
| pages | 9 |
| Iconographies | 4 |
| Vidéos | 0 |
| Autres | 0 |
Graphical abstract |
Highlights |
• | Developed human cytochrome P4503A4 gene with improved activity using in silico techniques. |
• | Synthetic gene was successfully cloned and expressed in probiotic yeast isolate S. cerevisiae. |
• | Recombinant S. cerevisiae (MTCC 25158) expressing human CYP3A4 showed an activity of 108 IU/mL. |
• | Degradation of leftover drug residues of amiodarone was observed as 66.32 % and 72.61 % in vitro and in vivo, respectively by recombinant S. cerevisiae. |
Abstract |
Background |
Amiodarone represents a principal antiarrhythmic pharmaceutical drug available in the market for the treatment of ventricular arrhythmias. However, despite its better efficacy, the usage of amiodarone is associated with extracardiac toxicity. The human body evolved a system of cytochrome P450 enzymes which play an essential role in the metabolism of harmful foreign substances. Therefore, CYP enzymes may either lead to the elimination or degradation of the leftover drug residues.
Objective |
The present study focused on successful utilization of Saccharomyces cerevisiae strain OBS2 with probiotic- cum- therapeutic potential and expressing in silico enhanced human cytochrome P4503A4 for the degradation of leftover drug residues of amiodarone in vitro and in vivo.
Methodology |
In this study, cytochrome P4503A4 (1W0E) was taken as a template and the predicted 3D model of mutant CYP3A4 was developed using different bioinformatics tools. Selected mutant (Glu165Asp) protein was reverse translated and transcribed into cDNA sequence. The cDNA of CYP3A4 was synthesized, cloned into p427TEF construct and transformed into Saccharomyces cerevisiae OBS2. The degradation of leftover drug residues of amiodarone in vitro and in vivo using recombinant Saccharomyces cerevisiae OBS2 expressing CYP3A4 was evaluated.
Result |
The CYP3A4 activity in recombinant probiotic yeast was observed as 108 IU/mL and in vitro degradation of leftover drug residues of amiodarone was observed as 66.32 %. Whereas, in vivo degradation of leftover drug residues of amiodarone was observed as 72.61 % along with recovery of organ damage in histopathological studies of the animal model.
Conclusion |
Saccharomyces cerevisiae OBS2 expressing CYP3A4 can be used for probiotic and therapeutic applications.
Le texte complet de cet article est disponible en PDF.Keywords : Amiodarone, Cytochrome P450 3A4, Saccharomyces cerevisiae, Docking simulation, Molecular cloning, Gene expression, Cardiovascular disease (CVD)
Plan
Vol 127
Article 110128- juillet 2020 Retour au numéro
Article précédent
- Pananx notoginseng saponins attenuate CCL2-induced cognitive deficits in rats via anti-inflammation and anti-apoptosis effects that involve suppressing over-activation of NMDA receptors
- Yi-jun Zhou, Jian-min Chen, Kiran Sapkota, Jiang-yi Long, Yuan-jun Liao, Jun-jun Jiang, Bing-yu Liang, Jin-bin Wei, Yan Zhou
- The gut microbiota confers protection in the CNS against neurodegeneration induced by manganism
- Hui Wang, Shidong Zhang, Feng Yang, Ruihua Xin, Shengyi Wang, Dongan Cui, Yan Sun
Bienvenue sur EM-consulte, la référence des professionnels de santé.
L’accès au texte intégral de cet article nécessite un abonnement.
Bienvenue sur EM-consulte, la référence des professionnels de santé.
L’achat d’article à l’unité est indisponible à l’heure actuelle.
Déjà abonné à cette revue ?