Drug – bio-affecting and body treating compositions – Plant material or plant extract of undetermined constitution... – Containing or obtained from a fruit – including berry
Reexamination Certificate
2001-02-05
2002-08-06
Tate, Christopher R. (Department: 1651)
Drug, bio-affecting and body treating compositions
Plant material or plant extract of undetermined constitution...
Containing or obtained from a fruit , including berry
C424S725000, C514S866000
Reexamination Certificate
active
06428825
ABSTRACT:
BACKGROUND OF THE INVENTION
1. Field of the Invention
The present invention relates to an herbal therapeutic product for controlling diabetes mellitus comprising at least one hypoglycemic compound extracted from the pulp of a fruit from a species of genus Eugenia specifically
Eugenia jambolana
and a process for the preparation of the same.
An embodiment of the invention resides in the various compounds of the product and their mixture having immediate as well as long lasting effects in controlling diabetes mellitus.
Another embodiment of the invention resides in the four active hypoglycemic compounds comprising the product of the subject invention. These four active hypoglycemic compounds individually have varying degrees of hypoglycemic characteristics, however, the combination has been found to be most effective.
Still another embodiment of the invention resides in the discontinuity of such herbal therapeutic product for a predetermined period after controlling the diabetes of the patients as the hypoglycemic compounds of such products are having the property to control the glucose level, for longer durations.
It has been found that the intake of doses can be postponed for 2-3 days after controlling the glucose level because of the long lasting effects of these hypoglycemic compounds.
The source of carbohydrates in the diet has a significant influence on the lipid(fat) metabolism in human being. An amount of sucrose is increased and the quantity of complex carbohydrates is decreased, the concentration of cholesterol and more particularly of triglycerides increases which has a correlation between blood lipid concentration and impaired glucose tolerance.
One of the objects of the present invention is that the product of the present invention is a 100% herbal product without any side effects.
Another object of the present invention is to treat the defect in metabolism of a patient suffering from diabetes, by increasing the activity of the glucose utilizing enzymes in the liver, muscles and adipose tissues.
The present invention relates to the product obtained from the species of genus Eugenia having hypoglycemic effects in the blood sugar level.
2. Description of Related Art
Kelkar in his article XP-000940531,
Phytomedicine,
Volume 3 (4), pages 353-359, 1996/97, teaches a purification of antidiabetic compounds from
Eugenia jambolana
fruit-pulp; proteolytic resistance and other properties.
The compounds have been identified as a peptidylglycan and an oligosaccharide with molecular weights of 6.0 and 1.2 kD, respectively. The binding between the sugars and the peptide in the peptidylglycan appears to be covalent. The amino acid and sugar composition of the peptidylglycan have been determined as having the sugars of the oligosaccharide. The intrinsic color of the peptidylglycan was attributed to the compound itself and not to the presence of pigment or metal. The peptidylglycan was resistant to degradation by proteolytic enzymes in vitro, explaining its efficacy in oral feeding.
Kelkar further teaches separation of hypoglycemic compound by molecular sieving (depending upon the size of two molecular). In short the purification process described Kelkar involves ultrafiltration, gel filtration, paper chromatography and SDS polyacrylamide tube gel electrophoresis.
Kelkar also teaches that hypoglycemic activity was found at two distinct positions eluting at 35-45 ml & 60-75 ml, which is not very precise, since vol. of fractions vary with the amount of loading, width and size of column and height of packed material.
Saigeeta Achrekar et al. in its review article XP000608379 teaches Hypoglycemic Activity of
Eugenia jambolana
and
Ficus bengalensis:
BRIEF SUMMARY OF THE INVENTION
Accordingly, the present invention relates to a herbal therapeutic product for controlling diabetes mellitus comprising at least one hypoglycemic compound extracted from the pulp of a species of the genus Eugenia, specifically
Eugenia jambolana.
The process for the preparation of the herbal therapeutic product to control the glucose level, comprises cleaning and drying the fruit of a species of family Eugenia to remove extraneous material from the outermost layer of the said fruit. De-seeding the fruit and soaking the said de-seeded fruit in water under controlled cooled conditions overnight to retain the activity of hypoglycemic compounds in the mixture of water and pulp of de-seeded fruit. It is followed by mixing the mixture to achieve a uniform consistency.
DETAILED DESCRIPTION OF THE INVENTION
The process for the preparation of such herbal product comprising cleaning and drying the fruit of a species of genus Eugenia of family Myrtaceae to remove the extraneous material from the outer layer of the fruit, deseeding the fruit, soaking the de-seeded pulp in water, mixing the mixture of water and pulp and keeping it overnight in a controlled cooled conditions to maintain the activity of hypoglycemic compounds, filtering the mixture in a conventional manner, washing the residue with water to extract all the active compounds from the mixture of water and pulp in the form of clear watery solution, reducing the water content by keeping the clear watery solution in controlled freezing conditions by conventional means to obtain the residues in a concentrated form, purifying the concentrated product in a chromatography by using slurries of different columns, and adding buffers in the purified mixture present in the chromatography to elute the mixture of hypoglycemic compounds.
The various steps involved in the aforementioned process are:
(a) The fruit of
Eugenia jambolana
is cleaned and dried to remove the extraneous material from the outer layer of the fruit;
(b) The seeds are removed from the fruit in a conventional manner and kept separately;
(c) The resulting de-seeded pulp is soaked in (distilled) water and the same is mixed to obtain a uniform consistency of the liquid;
(d) The mixture is kept overnight in a controlled cooled condition preferably between 4-10 degrees centigrade to maintain the activity of the hypoglycemic compounds present in the fruit pulp;
(e) The mixture is filtered in a conventional manner;
(f) The residue is washed with water to extract all active compounds from the pulp in the form of clear watery solution;
(g) The water content of the solution is reduced by keeping the clear watery solution in freezing conditions by conventional means to concentrate the residue;
(h) The concentrated product is purified by treating the same with slurries of solvents selected preferably from diethyl amino ethyl cellulose, anion exchange resins or silica gel and distilled water in the chromatography; and
(i) Buffers are added in the purified mixture present in the chromatography to elute the hypoglycemic compounds from the mixture.
The water content from said mixture can be reduced optionally to get the end product in solid form. The water is extracted from the mixture after filtration by means of lypholizer.
The pulp of
Eugenia jambolana,
is subjected to column chromatography for a period of 15 minutes to 2 hours to obtain a hypoglycemic fraction at a slightly acidic pH of 5-7, preferably 6.0 to 6.5.
The hypoglycemic fraction is subjected to thin layer chromatography using silica gel to form at least 2 visible bands. The R
f
value of all the four bands is between 0.1-1.0, wherein the R
f
value of one visible band which has a highly active hypoglycemic compound is between 0.5 to 0.7 and the R
f
value of the less active hypoglycemic compounds is between 0.1-0.2 and 0.9-1.0.
The pH of the column containing the mixture of chemical compounds is maintained at 6.0-8.0, preferably 6.5 to 7.5.
The buffer used in the chromatography is selected preferably from a phosphate buffer or citrate buffer.
The pH of the mixture of hypoglycemic compounds is to be maintained neutral.
The mixture of hypoglycemic compounds were further analyzed to separate the active hypoglycemic compounds by means of thin layer chromatography to separate the different compounds in the form of separate bands. Such bands were furt
Murthy Pothapragada Suryanarayana
Nasir Afreena
Prabhu Krishan Madhav
Sharma Suman Bala
Indian Council of Medical Research
Ladas & Parry
Tate Christopher R.
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