A PROCESS FOR THE CHARACTERIZATION OF THE BIOACTIVE COMPONENT PRESENT IN BARK AND ROOT OF AZADIRACHTA INDICA (NEEM) USED IN HERBAL REMEDIES FOR THE TREATMENT OF DIABETIC

摘要

Hypoglycemic activity of Azadirachta indica(Neem ) which is used in the treatment of diabetics is known. But bioactive component responsible for hypoglycemic activity remained unknown. The invention characterizes the bioactivity, present in alcoholic extract or in low molecular weight fraction (5000 dalton ) of aqueous extract of bark and root of the plant , reversibly inhibiting maltase and sucrase enzyme activities. In the digestion of starch, pancreatic amylase produces maltose from starch . Maltose as well as any sucrose if taken, are hydrolysed at the epithelium of the small intestine by membrane-bound maltase and sucrase into glucose or glucose and fructose.. Many low molecular weight maltase and sucrase inhibitors (acarbose, miglitol, valiolamine derivative) are clinically used for the treatment of type-II or insulin-independent diabetes Therapy is achieved by delaying glucose absorption into the blood stream due to inhibition of epithelial sucrase and maltase activities. The present invention characterized similar activity present in Azadirachta indica(Neem which could control inflow of glucose into blood after starch digestion. The active inhibitors molecule (s) may be a potential ant-diabetic drug or a lead molecule for future drug development. The present invention provides a process for the characterization of inhibitory activities of aqueous or alcoholic extract of Azadirachta indica root or bark on maltase and sucrase activities, which comprise collection of fresh or dried uninfected bark or roots of Azadirachta indica, washing them with water, blending in blender to paste, separating the aqueous solution by pressure filtration, further filtering the solution by ultrafiltration membrane of 5,000 dalton cut, collecting the filtrate and drying the solution in a lyophilizer, and characterizing the inhibitory activities of the fraction on activities of maltase and sucrase or extracting hand milled powder of root and bark each separately with ethyl alcohol for 24 hours, collecting the filtrates, removing alcohol by evaporation of the solvent 50°C and characterizing the inhibitory activities of dried powder on maltase and sucrase. In typical experiments, aqueous extraction gave 2.2 and 2 grams of solid from 100gm of dry bark and root respectively and ethanol extraction gave 2 and 1.4 grams of solid from 100 grams of dry bark and root respectively. Ki and IC50 values of the solids on maltase and sucrase of both mammalian and microbial origins were determined.