The in vitro stability and proteolytic enzyme of synthesized protamine-analog peptide R15
10.13220/j.cnki.jipr.2018.04.009
- Author:
Yong-Le ZHU
1
Author Information
1. Taishan Medical University
- Publication Type:Journal Article
- Keywords:
In vitro stability;
Metal carboxypeptidase;
Protease inhibitor
- From:
Journal of International Pharmaceutical Research
2018;45(4):295-300
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To establish an ultra performance liquid chromatography(UPLC)method for determination of prot- amine peptide analogul(R15),and investigate the metabolism in biological samples of R15 in vitro by the method. Methods: Chro- matographic separation was performed by pumping the mobile phase into a BioBasic C 18 (2.1 mm×100 mm,5 μm). The mobile phase was composed of 0.05% trifluoroacetic acid-water and 0.05% trifluoroacetic acid acetonitrile(98:2,V/V),using 10% trifluoroacetic ac- id-acetonitrile-water as precipitant. The stability of R15 in rat whole blood,liver and kidney homogenate was studied. Results: The in- tra-and inter-batch relative standard deviations (RSD)were less than 15%,the relative errors (RE)were in the range of ±15%. The range of calibration curve was 25-1000 μg/ml with a good correlation coefficient. The extraction recoveries of R15 were 99.43-100.43%. R15 was stable under various experimental storage conditions. Half-lives of R15 in rat liver,rat kidneys,rat blood,beagle dog blood and human blood were 27.4 min,9.1 min,15.6 min,11.9 min and 21.0 min,respectively. Compared with the control group without inhibi- tor,the inhibitory rates of EDTA,pepstatin and benzamidine HCl protease inhibitor in the experimental group was 92.7%,41.3% and 33.0%,respectively. Protamine analogue peptide was mainly metabolized by metal carboxyl peptidases. Compared with the control group,AEBSF,leupeptin and trypsin protease inhibitors had significant difference,but the inhibitory rate was low. Conclusion: The method is specific,sensitive,accurate and reliable. The metabolism of R15 in rat kidney is the fastest. The degradation of R15 in whole blood is possibly related to metal carboxypeptidase.
