Screening Antidepressive Effect Of MT And Quality Control Study

Abstract:The novel melatonin agonist, whose unique mechanism is combined with melatonin receptor (MT) and improves the symptoms of depression showing fewer adverse reactions, regulating circadian rhythm especially for improving sleep quality, is considered as a breakthrough in the field of antidepressants. In this study, the model of depression have been established to study the antidepressive effect of melatonin’s derivatives and the quality control of antidepressant M1.1. Screening of antidepressive effect of MT derivativesThe mice model with chronic stress depression was made by the chronic unpredictable stress and solitary custody. The changes of behaviors in mice were observed by determining immobility time in the tail suspension test, immobility time in the forced swimming test and locomotor activity in the open field test. Nine MT derivatives, including M1-M7and L1-L2, were screened and M1was studied the relationship between antidepressive effect and dose. Results:M1, M4, M7, L1and L2can improve the behaviour of depressed mice:decreasing the immobility time of mice in the suspension test (TST) and the forced swimming test (FST) and increasing locomotor activity in the open field test. Especially, antidepressant effect of M1have good relationship with its dose.2. Study on the quality control of drug M1Based on the screening antidepressant effect of MT, the quality control of the antidepressant Ml have been studied. The thesis mainly includes a system research on quality standards of M1and stability study of M1. The main content of the subject is as follow:2.1Confirmation of the structure of M1Analysis technologies such as UV, IR, MS,1H-NMR,13C-NMR, HMBC were adopted to analyze and identify the structure of synthetic target compound and confirm the correct structure of the samples. 2.2The determination content of M1by HPLCThe determination content of M1by HPLC have been established The method of determination the content of the M1is as follow:Chromatographic conditions:chromatographic column was Diamonsil C18(250mmx4.6mm,5μm); the mobile phase was methanol acetonitrile-water (43:12:45); The ultraviolet detection wavelength was224nm; the flow rate was1.0ml/min; the sample concentration was0.08mg/ml; the injection quantity was20μl and the column temperature was at room temperature. According to the results of methodological investigation such as sensitivity, precision, repeatability, linear range and so on, this method for the HPLC determination of M1content meets the requirements of the drug quality standard.2.3Analysis of the structure of one of related substances in M1The structure of related substances3have been confirmed by LC-MS, prepared it and determined its structure by UV、IR、MS、1H-NMR、13C-NMR、HMBC and NOE. Further study of other related substances of Ml is under going.2.4Establishment of the method for the analysis of related substances in M1Based on the study of quality control of MT, we have established a method for the analysis of the related substances in M1.Chromatographic conditions:chromatographic column was Diamonsil Ci8(250mm×4.6mm,5μm); Mobile phase was methanol-acetonitrile-water(43:12:45); The ultraviolet detection wavelength was224nm; the flow rate was1.0ml/min; the sample concentration was0.4mg/ml; the injection quantity was20μl and the column temperature was room temperature.Destructive test which evaluated the effect of strong acid, strong base and strong oxidation for M1was investigated. The experimental results have indicated that the effect of strong acid, oxidation were weaker. The method can well separate M1from its related substances. The method of determination of the related substances was self-control method of the principal component of the related substance.2.5Study stability testing of M1 Stability study is one of the main content of pharmaceutical quality control, closely relating to the drug quality research and the establishment of quality standards. The stability of M1was studied by exposing to high temperature, high humidity and strong light. The results have indicated that M1was stable under the condition of high temperature, high humidity and strong light.2.6SummaryBased on the confirmation of the structure of the samples, the method for the analysis of M1and its related substances had been established. We can indicate that the method was sensitive, accurate, repetitive and specifical by studying the method for the analysis of M1.Two aspects of the quality investigation and the stability study of M1that provide the basic data for the quality specification establishment of M1.