Title of Invention	A SYNERGISTIC PHARMACEUTICAL COMPOSITION FOR DIABETIC NEPHROPATHY
Abstract	The present invention provides a synergistic pharmaceutical composition comprising butyric acid, wheat fibre bran and guar gum for the treatment of diabetic nephropathy. The composition of the present invention is useful in reducing the albumin excretion and ameliorates the Glomerular Filtration rate.

Full Text

The present invention relates to a synergistic pharmaceutical composition for diabetic Nephropathy. Diabetic Nephropathy is one of the complications of diabetes meilitus involving basement membrane thickening as a result of reduction in Heparan Sulfate, Laminin and increase in type-IV Collagen/ Diabetic Nephropathy is characterized by increased excretion of albumin in the urine as a result of Glomerular Basement Membrane damage. Glomeruli become more porous to passage of macromolecules. During Diabetic Nephropathy the functional unit of kidney, the Nephrons are damaged leading to increased filtration rate and also increased excretion of proteins in urine. Therefore, compositions helpful in modulating the damaged kidney membrane have a tremendous scope in preventing the progression of Diabetic Nephropathic state. Diet plays a major role in management of diabetic complications. Dietary fibre - unabsorbable carbohydrates in the diet has many beneficial effects including slowing macromolecular digestion, slow release and absorption of glucose etc. The dietary fibres are fermented by microbes in the colon to short chain fatty acids and the role of butyric acid-a four carbon fatty acid in particular, on various physiological functions is receiving great attention. A reference can be made to patent number US6248375 wherein designed solid matrix nutritional product is designed containing dietary fibre and indigestible oligosaccharides to be administered to diabetics. The draw back of this nutritional product is that it does not include butyric acid, which is recognized for its role at the molecular level. A reference can be made to patent number US4598081 wherein a butyric acid analogue has been recommended for the treatment of complications of diabetes mellitus. The draw back is non-usage of dietary fibre, which acts as a reservoir of butyric acid and has supplementary beneficial effect. A reference can be made to patent number US6303586 wherein a supportive therapy in the form of stabilized rice bran is given in the treatment of diabetes. This includes both the solubilised and insolubilised fractions of the stabilized rice bran. The draw back of this therapy is that it includes only a single type of dietary fibre (rice bran). A reference may be made to patent number US6303174 that concerns with the food composition including resistant starch. The draw back of this composition is that it can include only a soluble fibre from grains and legumes. A reference can be made to patent number US6248390 wherein fibre-water-water containing soluble fibre is invented. The draw back of this invention is that it includes only water-soluble fibre. Hence, a pharmaceutical composition comprising butyric acid, wheat bran and guar gum has a significant role in ameliorating the increased Glomerular Filtration Rate and protein in urine and also prevent the progression of Diabetic Nephropathy. The main object of the present investigation is to provide a pharmaceutical composition comprising butyric acid, wheat bran and guar gum for treating Diabetic Nephropathy. An object of the present invention is to provide a pharmaceutical composition to reduce albumin excretion and ameliorate the Glomerular Filtration Rate. Yet another object of the present invention is to provide a pharmaceutical composition that provides a constant reservoir of butyric acid. Further object of the present invention is to provide a soluble fibre guar, which is a hypoglycemic agent. Accordingly the present invention provides a synergistic pharmaceutical composition useful for diabetic nephropathy comprising butyric acid, wheat bran and guar gum. The present invention also provides a method for the treatment of diabetic nephropathy. Accordingly, the present invention provides a synergistic pharmaceutical composition for diabetic nephropathy comprising: [a] butyric acid in the range of 0.025% to 0.075%; [b] an insoluble wheat fibre bran in the range of 1.0 to 5.0%; [c] guar gum fibre in the range of 1.0 to 2.5% as a soluble hypoglycemic agent; [d] optionally along with pharmaceutically acceptable additives. An embodiment of the present invention, wherein the preferred dosage of said composition is in the range of 250-750 mg/kg body weight. Yet another embodiment of the present invention, wherein the most preferred dosage of said composition is 500 mg/kg body weight. Still another embodiment of the present invention, wherein the sustained release of butyric acid of said composition is effective in the treatment of diabetic nephropathy. Further embodiment of the present invention, wherein the insoluble fibre wheat bran of said composition gets fermented all along the intestinal tract and acts a reservoir of butyric acid. Yet another embodiment of the present invention, wherein said composition is effective in reducing Glomerular Filtration Rate. Still another embodiment of the present invention, wherein said composition is effective in reducing urinary protein excretion. Yet another embodiment of the present invention, wherein said composition is used for the prevention, treatment and control of diabetic nephropathy. Further another embodiment of the present invention, wherein the pharmaceutically accepted additives are selected from group consisting of stearates and carbonates of magnesium, calcium and potassium. Yet another embodiment of the present invention, wherein the subject is selected from mammals. The following examples are given by the way of illustration of the present invention and therefore should not be construed to limit the scope of the present invention Example 1 Diabetes is induced in Male Wistar rats weighing between 120 g using streptozotocin at 55 mg/kg body weight and the rats are fed with wheat bran (1 to 5%), guar gum (1 to 2.5%) and butyric acid in the range of 1,2,4,6,8 and 10 mg/kg body weight/day in the diet. After 4 days measurement of fasting blood glucose level in the plasma and urinary sugar levels assessed the diabetic status. Fasting blood glucose level is measured in the plasma collected from retro-orbital plexus of the rats by glucose oxidase method. The reaction mixture is incubated for 15 minutes and absorbance is measured at 520 nm. Urinary sugar levels are measured by Dinitrosalicylic acid method. The reaction mixture is kept in boiling water bath for 10 minutes and the colour developed is measured at 540 nm. Urinary creatinine is estimated by alkaline picrate reagent method. The reaction mixture is incubated at room temperature for 15 minutes and absorbance is measured at 520 nm. The plasma is deproteinsed using tungstic acid prior to creatinine estimation. The Glomerular Filtration Rate is determined at the end of fifth week by measuring urinary creatinine and plasma creatinine . The formula used is Urinary creatinine (mg/dL) X Urine volume (ml) X (FORMULA REMOVED) Urinary protein is determined at the end of fifth week by TCA precipitation method and the absorbance is measured at 660 nm . The increased levels of fasting blood glucose, urinary sugar and urinary volume are reduced to a considerable extent. The progression of Diabetic Nephropathic state is considerably prevented by reducing the Glomerular Filtration Rate and urinary protein with the composition involving butyric acid, wheat bran and guar gum. Example 2 Diabetes is induced in Male Wistar rats weighing between 120 g using streptozotocin at 55 mg/kg body weight and the rats are fed with wheat bran (1 to 5%), guar gum (1 to 2.5%) and butyric acid in the range of 1,2,4,6,8 and 10 mg/kg body weight/day in drinking water. After 4 days measuring fasting blood glucose level in the plasma and urinary sugar levels assessed the diabetic status. Fasting blood glucose level is measured in the plasma collected from retro-orbital plexus of the rats by glucose oxidase method. The reaction mixture is incubated for 15 minutes and absorbance is measured at 520 nm. Urinary sugar levels are measured by Dinitrosalicylic acid method. The reaction mixture is kept in boiling water bath for 10 minutes and the colour developed is measured at 540 nm. Urinary creatinine is estimated by alkaline picrate reagent method. The reaction mixture is incubated at room temperature for 15 minutes and absorbance is measured at 520 nm. The plasma is deproteinsed using tungstic acid prior to creatinine estimation. The Glomerular Filtration Rate is determined at the end of fifth week by measuring urinary creatinine and plasma creatinine . The formula used is (FORMULA REMOVED) Plasma creatinine (mg/dL) X Body weight (g) X 1440 (min) Urinary protein is determined at the end of fifth week by TCA precipitation method and the absorbance is measured at 660 nm. The increased levels of fasting blood glucose, urinary sugar and urinary volume are reduced to a considerable extent. The progression of Diabetic Nephropathic state is considerably prevented by reducing the Glomerular Filtration Rate and urinary protein with the composition involving butyric acid, wheat bran and guar gum. Example 3 Diabetes is induced in Male Wistar rats weighing between 120 g using streptozotocin at 55 mg/kg body weight and the rats are fed with wheat bran (1 to 5%), guar gum (1 to 2.5%) and butyric acid in the range of 250,500 and 750 mg/kg body weight/day in the diet. After 4 days measuring fasting blood glucose level in the plasma and urinary sugar levels assessed the diabetic status. Fasting blood glucose level is measured in the plasma collected from retro-orbital plexus of the rats by glucose oxidase method. The reaction mixture is incubated for 15 minutes and absorbance is measured at 520 nm. Urinary sugar levels are measured by Dinitrosalicylic acid method. The reaction mixture is kept in boiling water bath for 10 minutes and the colour developed is measured at 540 nm. Urinary creatinine is estimated by alkaline picrate reagent method. The reaction mixture is incubated at room temperature for 15 minutes and absorbance is measured at 520 nm. The plasma is deproteinsed using tungstic acid prior to creatinine estimation. The Glomerular Filtration Rate is determined at the end of fifth week by measuring urinary creatinine and plasma creatinine . The formula used is (FORMULA REMOVED) Plasma creatinine (mg/dL) X Body weight (g) X 1440 (min) Urinary protein is determined at the end of fifth week by TCA precipitation method and the absorbance is measured at 660 nm . The increased levels of fasting blood glucose, urinary sugar and urinary volume are reduced to a considerable extent. The progression of Diabetic Nephropathic state is considerably prevented by reducing the Glomerular Filtration Rate and urinary protein with the composition involving butyric acid, wheat bran and guar gum. Example 4 Diabetes is induced in Male Wistar rats weighing between 120 g using streptozotocin at 55 mg/kg body weight and the rats are fed with wheat bran (1 to 5%), guar gum (1 to 2.5%) and butyric acid in the range of 250, 500 and 750 mg/kg body weight/day in drinking water. After 4 days measuring fasting blood glucose level in the plasma and urinary sugar levels assessed the diabetic status. Fasting blood glucose level is measured in the plasma collected from retro-orbital plexus of the rats by glucose oxidase method. The reaction mixture is incubated for 15 minutes and absorbance is measured at 520 nm. Urinary sugar levels are measured by Dinitrosalicylic acid method. The reaction mixture is kept in boiling water bath for 10 minutes and the colour developed is measured at 540 nm. Urinary creatinine is estimated by alkaline picrate reagent method. The reaction mixture is incubated at room temperature for 15 minutes and absorbance is measured at 520 nm. The plasma is deproteinsed using tungstic acid prior to creatinine estimation. The Glomerular Filtration Rate is determined at the end of fifth week by measuring urinary creatinine and plasma creatinine (Table 1). Table 1 Effect of butyric acid on Glomerular filtration Rate in control and diabetic rats (Table Removed) Groups with numbers (eg. SFD-250) represent mg of butyric acid fed/kg body weight/day SFC - Starch fed control SFD - Starch fed diabetic FFC - Fibre fed control FFD - Fibre fed diabetic Values are mean ± SEM of 6 rats in control and 14 rats in diabetic groups. Statistically significant compared to SFC at p The formula used is (FORMULA REMOVED) Plasma creatinine (mg/dL) X Body weight (g) X 1440 (min)rinary protein is determined at the end of fifth week by TCA precipitation method and the absorbance is measured at 660 nm (Table 2). Table 2 Effect of butyric acid on urinary protein excretion in control and diabetic rats (Table Removed) Abbreviations as in Table 1 The increased levels of fasting blood glucose, urinary sugar and urinary volume are reduced to a great extent. The progression of Diabetic Nephropathic state is significantly prevented by reducing the Glomerular Filtration Rate and urinary protein with the composition involving butyric acid, wheat bran and guar gum. The advantage of the invention are as follows 1. A synergistic pharmaceutical composition for treating Diabetic Nephropathy helps in preventing complications of diabetes. 2. A synergistic pharmaceutical composition for treating Diabetic Nephropathy by reducing the Glomerular Filtration Rate. 3. A synergistic pharmaceutical composition for treating Diabetic Nephropathy by reducing the urinary protein excretion. We Claim: 1. A synergistic pharmaceutical composition for diabetic nephropathy comprising: [a] butyric acid in the range of 0.025% to 0.075%; [b] an insoluble wheat fibre bran in the range of 1.0 to 5.0%; [c] guar gum fibre in the range of 1.0 to 2.5% as a soluble hypoglycemic agent; [d] optionally along with pharmaceutically acceptable additives. 2. A synergistic pharmaceutical composition for diabetic nephropathy substantially as herein described with reference to the foregoing exmples.

Documents:

305-DEL-2002-Abstract-19-05-2008.pdf

305-del-2002-abstract.pdf

305-DEL-2002-Claims-19-05-2008.pdf

305-del-2002-claims.pdf

305-del-2002-correspondence-others-1.pdf

305-DEL-2002-Correspondence-Others-19-05-2008.pdf

305-DEL-2002-Correspondence-Others.pdf

305-DEL-2002-Correspondence-PO.pdf

305-del-2002-description (complete)-19-05-2008.pdf

305-del-2002-description (complete).pdf

305-DEL-2002-Form-1-19-05-2008.pdf

305-del-2002-form-1.pdf

305-del-2002-form-18-1.pdf

305-DEL-2002-Form-18.pdf

305-DEL-2002-Form-2-19-05-2008.pdf

305-del-2002-form-2.pdf

305-DEL-2002-Form-3-19-05-2008.pdf

305-del-2002-form-3.pdf