WO2005048915A2 - Renoprotective and lipid lowering oral compositions - Google Patents

Abstract

The present invention discloses a renoprotective and lipid lowering herbal oral composition comprising granules of Hemidesmus indicus along with pharmaceutically acceptable excipients. Further the invention discloses the use of such herbal composition in treatment of primary glomerulonephritis and/or aminoglycoside induced nephrotoxicity and method of treatment for the same.

Description

Related Application

This application claims priority from India National patent application serial No.

1003/MUM/2003, filed 24^th Sept 03.

Technical Field

The present invention relates to renoprotective and lipid lowering oral solid dosage composition developed from Hemidesmus indicus Linn Roots.

Background and Prior Art

Renal diseases have always been a challenging field of medicine. They continue to attract the interest of successive generations of physicians. Definitive therapeutic strategies for various renal disorders are still under investigation. The end stage renal disease in the form of failure causes significant morbidity and mortality. However, its treatment (dialysis and transplantation) remains out of reach for a vast majority of Indian patients because of high cost. Renal failure can also be due to nephrotoxins. Many clinically popular drugs such as amino glycoside antibiotics, amphoteracin B, radiolographic contrast agents e.g. sodium and meglumin salts of diatrizoate and isothalmate, analgesics, cyclosporin can cause deterioration of kidney function even when the dose has been adjusted properly for renal insufficiency. Drug induced nephrotoxicity ranges from mild dysfunction and electrolyte disturbances to acute tubular necrosis and chronic nephropathy. In acute renal dysfunction reestablishment of urine output with diuretics, reduction in intake of nitrogenous substances, water and electrolytes and maintaining adequate nutrition are the main therapeutic measures. Several clinical and experimental strategies have been employed in order to ameliorate or abolish signs of nephrotoxicity. Most of these are still need to elucidate the mechanism of action of the drug nephrotoxicity and to discover safe, practical and effective agents to ameliorate the nephrotoxicity in patients at high risk. Attempts have been made to introduce a specific agent, which can alter the course of renal damage and reorient the existing symptomatic and supportive management to more definitive form. Cholesterol is a lipid substance that is used for many body processes. Low-density Hpoprotein (LDL) is known as the bad cholesterol, is the main source of build up on the blood vessel walls. High density hpoprotein (HDL) cholesterol carries cholesterol back to the liver and helps prevent build upon the blood vessel walls. High cholesterol level is seen in patients who suffer from renal disease.

The most important hypolipidemic drugs available today belong to statin and fibrate classes. However theses therapeutic agents are not free from side effects. Statins (HMG- CoA reductase inhibitors) the most widely used drugs today which hitherto were thought to be very safe drugs, have exhibited side effects following long term therapy. The adverse effects, which have become the concern are increases in hepatic transminases and myopathies. The incidence of myopathy associated with rhabdomyolysis and renal failure is increased subsequent to such treatment. Also, the HMG-CoA inhibitors block mevalonate, which is a common precursor for all isoprenoids such as ubiquiones (Co-enyme Q-10), the dolichols, isopentenyl t-RNA etc. Therefore, long-term blockade of mevalonate synthesis leads to Q-10 deficiency. Serum Co-enzyme Q-10 is important for cardiac function. The commonest side effects of fibrates, particularly clofibrate therapy are gastrointestinal upsets including nausea, vomiting, diarrhea, dyspepsia, flatulence and abdominal discomfort. Elevated creatinine phosphokinase concentration during clofibrate therapy may be associated with a syndrome of muscle pain and weakness. Long-term studies have demonstrated an increased incidence of cholecystitis, gall stones and sometimes pancreatitis in patients receiving clofibrate and some studies have indicated cardiovascular disorders. The unexpected finding of an increased mortality rate in patients taking clofibrate in the WHO study produced serious concern over the long- term safety of clofibrate and ultimately led to its withdrawal in many countries [Oliver, M.F.et al; Lancet, ii, 600- 604(1984)]. The general therapeutic approach to patient with acute renal failure aims at diagnosing and correcting all treatable causes or renal dysfunction. The attempts are made to reestablish an adequate urine output including, taking diuretics like frusemide, conservative therapy includes a decreased intake of nitrogenous substances, water and electrolytes to match the output and providing adequate nutrition. Frequent clinical and biochemical monitoring of patients is necessary to readjust to the therapy. Patients with chronic renal failure are usually managed best with conservative measures, which include control of associated hypertension, reduction in protein diet, regulation of fluid intake, treatment of anemia acidosis, infections and uraemic osteodystrophy and neuropathy.

Apart from these routinely followed established therapeutic measures a variety of pharmacologic agents have been proposed to prevent acute renal failure or to attenuate its severity or hasten the recovery. Because of the lack of adequate scientific information, the use of these modalities has remained controversial. The details are as follows:

1) Mannitol: - The protective effect of mannitol in many experimental models of Acute Renal Failure has been attributed to increase in solute excretion, diminution of cell swelling, both contributing to prevention of tubule obstruction and vasodilation. However, these effects are partial and inconsistent. Mannitol is known to be an effective hydroxyl radical scavenger.

2) Calcium channel blocker: - Appreciation of the fact that cell injury is marked by an increase in cytosolic and mitochondrial calcium concentrations and these shifts in calcium may aggravate the degree of cellular injury in acute renal failure has lead to the use of calcium channel blockers.

3) Prostaglandin: PGI₂ has been used for its cytoprotective action. Since PGI₂ has a vasodilatory effect in the kidney the results suggest that it may play a specific role in maintaining normal renal blood flow and glomerular filtration rate during the aminoglycoside nephrotoxicty. The inhibition of prostaglandin production by non- steroid anti-inflammatory drugs prevents these compensatory mechanisms and worsens the renal damage.

4) Xanthine oxidase inhibitors:- AUopurinol and oxipurinol have been used to prevent the formation of reactive oxygen species during the conversion of hypoxanthine or xanthine to uric acid.

5) Free-radical scavengers: These agents scavenge the super oxide radical or limit its formation and prevent tissue damage has been tried.

Based on the mechanisms involved in the pathogenesis of chronic renal disease following measures may appear to be rational in slowing the natural loss of renal function: - 1. Control of systemic hypertension: Systemic hypertension is a regular feature of progressive renal insufficiency and elevation of blood pressure appears to accelerate loss of renal function. 2. However, another antihypertensive agent, metoprolol, does not prevent the progressive decline in renal function despite a prolonged therapy of 7 years. It has been observed that antihypertensives like calcium channel blockers and ACE inhibitors are more beneficial than the combination of reserpine, hydralazine and hydrochlothiazide also known as triple therapy. The former agents' posses ability to lower intraglomerular capillary pressure also while the others are only effective agents to control systemic hypertension. 3. Control of glomerular capillary hypertension: With the knowledge of these differential effects, focus has now shifted to lower the glomerular capillary pressure. The inadequate constriction of afferent arteriole allows the transmission of increased systemic pressure to glomerular capillary. ACE inhibitors cause preferential dilation of efferent arteriole, lowering intraglomerular pressure and thus reduce the renal damage. 4. Reduction in protein intake: - An increase in protein intake stimulates the secretion of glucagon, dopamine, vasopressin, eicosanides and rennin. Prospective randomized, controlled trials have been conducted in nondiabetic patients to study the effects of low protein diets in renal insufficiency. The Meta analysis of this data revealed that dietary protein restriction indeed delays the onset of end-stage renal disease. However, the dietary management in renal diseases is not easy, especially when long-term administration is necessary. A large multi centered clinical trial modification of diet in renal disease study 1992, currently underway will hopefully clarify the roles of dietary proteins restriction and aggressive reduction in blood pressure in the therapy of chronic renal disease in humans. 5. Reduction in dietary phosphate intake: - Phosphate restriction also reduces proteinuria and prostanoid production in several models of chronic renal diseases. It may also reduce the oxident stress, however such restriction has no effect on glomerular hypertension and lipid accumulation, the leading factors to progressive renal injury. 6. Phosphate binders: These agents also decrease phosphate intake and proposed mechanisms are same as above. 7. Control of hyperlipedemia: - It has been proposed that correction of hyperlipedemia may decrease proteinuria and / or slow the progression of renal failure. Lipid lowering agents such as lovastatin, primastatin, probucol, and nicotinic acid reductase have been tried. However none of these agents are without toxic effects. Especially lacking are long-term trials to determine safety and efficacy of drug therapy.

6. Corticosteroids: - Long term administration of corticosteroids leads to manifestation of adverse effects and in ISKDC trial large number of withdrawals resulted because of hypertension and encephelopathy induced by steroids. This renders the data generated insufficient to reach any conclusions. 7. Immunosuppressive therapy: - The cytokines are implicated in chronic renal damage. Hence various immunosuppressive therapies are being tried as a therapeutic strategy. The use of cytotoxic drugs is however not without hazards which limits their applicability. Cyclosporin has shown to induce remission in minimal change glomerulonephritis, however lasting remission following discontinuation of therapy are uncommon and prolonged therapy may be associated with side effects including nephrotoxicity.

8. Reno protective agents: - ACE inhibitors and calcium channel blockers are the two classes of anti-hypertensive agents with novel effects on renal function. In both experimental models and clinical studies in patients with essential hypertension, calcium channel blockers were found to increase the renal blood flow, glomerular filtration rate and urinary sodium excretion. Similar effects were observed with ACE inhibitors with respect to renal blood flow and glomerular filtration rate, but they were found to cause sodium retention. In addition in cases of renovascular hypertension, it was observed that ACE inhibitors can cause marked reductions in GFR. The mechanism for the reno protective action of ACE inhibitors is not clearly established. As mentioned earlier hyperfiltration and raised intraglomerular pressures are responsible for progression of renal disease. ACE inhibitors reduce glomerular pressure. Angiotensin II plays role of growth regulator and induces hypertrophy of vascular smooth muscles and hyperplastia of other mesangial and endothelial cells. It also activates several growth promoters as well as other prqtoncogenes. There is good evidence that ACE may be involved in the inflammatory process. Based on several series of clinical and experimental evidence of ACE inhibitors has been shown that ACE inhibitors are advantageous in patients with diabetic nephropathy. However with regard to progression of non-diabetic chronic renal disease, literature has shown that as yet it cannot be concluded that ACE inhibitors modify the rate at which the nephropathy progresses in non- diabetic patients and therefore may be of little help to these patients. Though calcium channel blockers like verapril have shown to be protective against gentamicin-induced nephrotoxicity; it has been reported that ditiazem enhances gentamicin nephrotoxicity. A similar result is observed in verapril that it increases the nephrotoxic potential of gentamicin in rats.

The rapid advances in the field of dialysis and transplant offer a new horizon in the therapeutics. Yet these modalities have their drawbacks, technical problems, infections, metabolic complications and hypertension are common complications in patients undergoing dialysis, further renal transplant is also not an answer. Apart from complications of major operations, medical problems of transplantation are rejection, adverse effects of steroid and immunosuppressive therapy, infections and restricted lifestyle. The major hurdle in India is the high cost of these sophisticated techniques.

The studies with pharmacological agents like calcium channel blockers and angiotensin converting enzyme inhibitors have raised a new hope. A "nephroprotective" role was claimed for their effects. However studies carried out on calcium channel blockers by Gomez et al 1989 and Bianchi et al 1991, could not prove these claims. Similar disappointment was also felt when ACE inhibitors were evaluated by Zucchelli et al 1992, and Mourad and coworkers (1993). On one hand ACE inhibitors have proved to be nephroprotective in diabetic nephropathy , but on the other hand , various clinical studies in non-diabetic diseases are inconclusive as yet. Thus their role as nephroprotectors not only requires to be confirmed, but hurdles of cost, side effects and contraindications also need to be overcome. These facts emphasize a need for a renoprotective agent, which would be safe, reasonable in cost and effective beyond doubt.

In recent times there has been renewed interest in herbal remedies and studies have been carried out to establish the clinical efficacy of such herbs. Search of Ayurvedic literature reveals that various plants like Boerhaavia diffusia (punarnava), Tribulus terrestris (gorkshur), Aerva lanata (Pashanabhed) etc. are used for renal disorders. There are many plants mentioned in Ayurvedic medicine, which are useful as diuretics as follows: l.Tinospora cordifolia (Guduchi) : Stem is used in dyspepsia, fevers and urinary diseases;

2. Tribulus terrestris (Gokshura) : it is useful in urolithiasis, , impotence and kidney dysfunction;

3. Boerhavia diffusa (Punarnava) : it is good for the kidneys, especially the nephron cells that are damaged by long-term hyperglycemia (diabetes);

4. Bacopa monnieri (Brahmi) : known as diuretic;

5. Rice ((Shali) : known as diuretic;

6. Rice (Vrihi) : known as diuretic; and

7. Flat beans (Nispara) : known as diuretic

The plants employed for cholesterol disorders are: -

1. Commniphora wightii (guggul): It lowers the level of cholesterol;

2. Terminalia arjuna (arjuna) : It reduces the level of triglycerides and cholesterol and has been reported to enhance the synthesis of LDL-apoprotein (apoB). This enhances the elimination of cholesterol from the body;

3. Courcuma longa (turmeric) : It reduces lipid peroxides, increases HDL Cholesterol, and decreases total serum cholesterol; and

4. Trigonella foenumgraecum (methi) : It lowers elevated cholesterol and triglyceride levels in the blood.

The above-mentioned herbal as well as allopathic drugs suffer from number of disadvantages. 1. Some of Allopathic drugs, which are used either for lowering of lipids or for kidney protection have quite a number of side effects; 2. In case of other herbs which are mentioned above have single effects i.e. either herb is useful in kidney disorders or then it is cholesterol lowering; 3. Adulteration of plant material is often there, therefore, the formulation, which does not contain standardized herb, does not show much efficacy; and 4. Very often the amount of base material is more compared to active components and thus the product is not very useful.

Hemidesmus Indicus Linn also called "Sariva" or " Anantamool" is used in traditional medicine in form of decoction, avaleha (syrup with or with out alcohol) in combination with other drugs to treat diseases. It has been traditionally (as per literature survey) used in various diseases like urinary disorders, fever etc. The plant has a variety called Hemidesmus Indicus Var Pubescens. Both the plants were used for study independently; Hemidesmus Indicus Linn is the preferred variety.

Hemidesmus indicus and it's variety, Hemidesmus indicus var pubescens, are found throughout India. The roots are cylindrical, thick and hard and somewhat tortuous, sparsely branched being provided with a few thin roots and secondary roots. External appearance is dark brown in color, sometimes with a violet grey tinge. The odor is agreeable resembling those of tonka beans and is due to the presence of coumarin. The taste is sweetish, though very slightly acid and aromatic.

The roots are bitter, sweet, astringent, aromatic, refrigerent, emollient, depurative, aphrodisiac, carminative, appetizer, antihelmenthic, alterant, demulcent, diaphoretic, febrifuge, expectorant and tonic ref -[Kirtikar KR, Basu BD, Indian Medicinal Plants ,Vol-III, L.M.Basu, Allahabad, p.1596, 1933]. It is also useful in vitiliated conditions of pitta, burning sensation, leucoderma, leprosy, skin diseases, pruritas, asthma, bronchhiatus, hyperdipsia, opthalmopathy, hemicrania, epileptic fits, dyspeptic fits, dyspepsia, helminthiasis, diarrhoea, dysentery, haemorrhoids, strangury, leucorrhoea, syphilis, abscess, arthralgia, fever and general debility. [Kirtikar K. R, Basu B. D., Indian Medicinal Plants ,Vol-III, L.M.Basu, Allahabad, p.1596, 1933, Nadkarni AN, Dr.K.M .Nadkarni's Indian Materia Medica, VOL- I: 619& 64, Popular Book Depot, Bombay 1954, Robert E, Vegetable Materia Medica of India & Ceylon, Plat Ltd, Colombo p. 201, 1931, Sanyal D. and Ghosh R. B., Vegetable Drugs of India , The medical publishers, Calcutta, p. 332, 1937].

Pharmacological study on different extracts of the plant has been described in the prior art. The steam distilled fraction appeared to be almost inactive pharmacologically [Satoskar RS, shah LG, Bhatt K and Sheth, U.K., Indian J Physiol Pharmacol, 1962, 6, 68]. Study on ethanolic extract was carried out to find the effect on respiration. [Dhar MX, Dhar M. M, Dhawan B. N, Mehrotra B.N, and Ray C, Indian J. Exp. Bio. 6 (1968), 232]. The petroleum ether, chloroform and alcoholic extracts of Hemidesmus indicus showed antibacterial activity against different microorganisms like S. albus, S. typhosa etc. [ICMR Bulletin, Request on Indian medicinal Plants, part-Ill, 1972, 2(6), 3].

The aqueous ethanolic extract of whole plant was tested for antibacterial, antifungal, antiprotozoal, antihelminthic, antispirochaetal hypoglycaemic and anticancer activities. [Dhar MX, Dhar M. M, Dhawan B. N, Mehrotra B.N, and Ray C, Indian Journal Exp. Bio. 6 (1968), 232]. The essential oil obtained from the plant exhibits marked antibacterial activity against B. proteus, P. aeruginosa, S. pyogenes and E. coli. [RPrakash et al, Indian Perfume, 1983, 27, 197-199].

The aqueous ethanolic extract of the whole plant (0.5 mg/ml) was tested for antiviral activity against Raniket disease virus (RDV) and vaccinia virus [Babber O.P, Chόwdhury BL, Singh, MP, Khan SK and Bajpai S, 1970, Indian J. Exp. Bio, 8, 304; O. P. Babber, B.Choudhary et al, Journal of Experimental biology, 1970, 8, 304-312; Babbar O, Joshi M.N, Indian journal Med. res, 1982, 76(supp)54-65]. In vitro evaluation of inhibitory nature of extracts of 18 plants species (including Hemidesmus indicus) against 3-keratinophilic fungi was studied [Qureshi S et al, Hindustan Anti Bull, 1997, 39(1-4), 56-60] The ethyl acetate extract of Hemidesmus indicus has been reported to exhibit significant anti-inflammatory activity. [Dutta M. K., Sen T. K. and Sikdar S., 1982, Proc Indian Pharmacol Soc. XIV, Ann Conf. Bombay, 31^st Dec, 1981, Indian J Pharmacol 14, 78].

Plant is noteworthy species used in stomach ulcer [Jain s.p, Etheno-medico-botanical survey of ambikapur, Fourth inter cong eteno bio, 1994, 293]. In another botanical survey it was interesting to note that the plants like Hemidesmus indicus, Thymofolia etc. in medicated form were used on scalp for strengthening the skull bone of children [Banerjee et al, Plants used by tribals, fourth inter cong ethenobio, 1994, 340].

Hemidesmus indicus was used in research carried out to see the effect of medicinal plants in management of tumors (carcinoma} [Ka nick C.R et al, Some observation on the use of Indian Medicinal plant in management of tumors, Arya Vaidya, 1997, 10 (4), 244-249]. Attempts were made to review the different sources of Ayurvedic drug Sariva, of four different taxa are being used in the name of Sariva,. [Nayar R.C, A review on the ayurvedic drug " Sariva" , Jour. Res. Ind. Med. Yoga & Homeo, 1979, 14: 2] . Diagnostic characters of drugs available under name "Sariva" are recorded in Short research Communication paper [Wahi A.K, Khosala RX, Mukherjee A.K, Journal. Res. Ind. Med. Yoga and Homeo, 1979, 14: 2]. The market sample of the crude drug under the local name Makali beru (kannada) has been identified as D. hamiltonii W. and A. and details are explained. [Togunashi VS, Venkateram Bsand Yoganarasimhan SN, Journal Research Indian Medical Yoga and Homeo, 13: 4. 1978].

Identification of Sariva using TLC for quick identification was carried out. [Togunashi VS, Venkateram Bsand Yoganarasimhan SN, Journal Research Indian Med Yoga and Homeo, 13: 4. 1978]. A number of characters of the drug have been enumerated by which it can be distinguished with root of Hemidesmus indicus [Wahi S.P and Ansari M. S., Journal research Indian Med., 1971, 5:2]. The utility of Jwaradhan Dashomani in treatment of fever was established. [Vyas Y. et. al., Utility of Jwaradhan Dashomani in treatment of fevers, Sachitra Ayurved, 1993, 5; 45; (10), 784-755] . The methanolic root extracts explored for neutralization of snake venom (Vipera russellii) activity. [Alam M. I. et al, Arya Vaidya, 1996, 10(4), 244-249]. One of the Botanical surveys revealed the use of Hemidesmus indicus and other plants in various ointments used for eye. [Sahoo et al, Plants used in ophthalmic drugs in Orissa, Glimpses of Indian Ethenopham, 1995, 173]. Physico-Chemical methods of Identification from finished products containing Hemidesmus indicus were carried out [Aravindakshan G., Ramiah N., Journal Science Research PI and Med., 1982, 3 (2-3), 57-60]. Terpenoids, B-sitosterol, pregnane glycosides, pregnane ester glycosides, indicusin- pregnane glycoside was also isolated from Hemidesmus indicus root powder [Madan M. et al, Phytochemistry, 1992, 31(2), 4036-4037, Ramesh Chatterjee et al, Journal Indian chemical Society, 1955, 32 (7), Chandra Khare et al, Pregnane glycosides, Pytochemistry, 1994, 35(6), 1545-1548, Kanchan Oberai, Anakshi Khare et al, 1985, 24, 2395-2397, Kamal Prakash et al, 1991, 30, 297-299, Deepak S et al, Natural product letter; 1995, 6(2), 81-86]. Hemidesmus indicus and other plants were screened for energy hydrocarbons and phytochemicals [Sekan, Bioresource- tec, 1998, 65(3) 257-259]. Hemidesmus indicus found to be protective against Rifamicin and Isoniazid induced hepatotoxicity in rat [Prabakan mookan, Fitoterapia, 2000, 71(1) 55-59].

Thus the present invention provides a non-synthetic, herbal therapeutic composition for protection of kidney and control of associated hypercholesterolemia (high cholesterol levels) in mammals.

Objectives

The main objective of the present invention is to provide herbal composition effective in protecting the kidneys and in lowering lipid levels without side effect associated with synthetic drugs. Another object of the present invention is to provide a herbal composition which is safe, has no mammalian toxicity and has maximum efficacy.

Summary of the invention

The present invention provides a renoprotective and lipid lowering herbal oral composition comprising granules of Hemidesmus indicus along with pharmaceutically acceptable excipients. The granules of Hemidesmus indicus comprise standardized root powder, diluent, dry starch and disintegrant. The granules comprise standardized root powder, dry starch powder, diluent such as Maize starch; disintegrant such as Colloidal Anhydrous Silica. The pharmaceutically acceptable excipients are selected from lubricant such as Magnesium stearate and moisture protective excipients selected from hydroxypropyl methyl cellulose in combination with microcrystalline cellulose.

The granules of Hemidesmus indicus are prepared by blending standardized root powder, dry starch, diluent, disintegrant and the blend obtained is either used as such in powder form or granulated by dry granulation. The granules are coated with moisture protective excipients and lubricated with lubricants such as Magnesium Stearate and filled into hard gelatin capsules.

The granules may be further compressed with pharmaceutically acceptable excipients into tablets and optionally coated with protective coating.

The herbal composition maybe in the form of powder filled into hard gelatin capsules or compressed into tablet by conventional technique.

The root powder is standardized in a way to comprise alkaloids 6.5% to 7.5%, fatty acids 3.7 % to 4.2% , tannins 0.2% to 0.3%, phenolics 1.1 % to 1.2 % ,acidic flavonoids 5.2% to 5.4 % and basic flavonoids 0.7 % to 0.8%; elements such as calcium 23.3% to 25.5%; magnesium 7.9% to 9.16%;chloride 0.008% to 0.016%;sulphate trace amount and iron 1.1% to 1.3% and total ash content 4.3% to 5.3%, acid insoluble ash content 13.3% to 15.1%, moisture content 8.7% to 9.1% and crude fiber content 15.1% to 19.5%.

The said composition is in the fonn of powder or granules. The powder or granules are filled into hard gelatin capsule.

The herbal composition is used to treat primary glomerulonephritis (biopsy proven) and/or aminoglycoside induced nephrotoxicity.

The herbal composition is useful in glomerular as well as tubular dysfunction.

Method of treatment comprising administering a therapeutically effective amount of said herbal composition to a subject in need suffering from primary glomerulonephritis

(biopsy proven) and/or aminoglycoside induced nephrotoxicity.

The administration of said herbal composition in (veterinary use) mammals increases

Hb level due to the presence of iron content.

The administration of said herbal composition in (veterinary use) mammals decreases serum triglyceride levels, serum creatinine, serum cholesterol, systolic and diastolic blood pressure.

The administration of said herbal composition in (veterinary use) mammals decreases in organ to body weight ratio particularly in case of kidney.

The administration of said herbal composition in (veterinary use) mammals increases in body weight of animals and an intake of food and water.

Detailed description:

The present invention provides a herbal composition useful for the protection of kidney and Heart and methods of using these compositions.

The herbal composition according to the present invention is particularly useful for the treatment of subjects suffering from kidney dysfunction from mild to moderate level due to aminoglycoside toxicity and kidney dysfunction due to primary glomerulonephritis. The present renoprotective and lipid lowering herbal oral composition comprises granules of Hemidesmus indicus alongwith pharmaceutically acceptable excipients. The granules of Hemidesmus indicus comprise root powder, binder, diluent and disintegrant.

The granules comprise root powder, binder such as dry starch powder, diluent such as Maize starch; disintegrant such as Colloidal Anhydrous Silica. The granules comprises 250mg root powder, binder such as dry starch powder, 50mg diluent such as Maize starch; and 3mg disintegrant such as Colloidal Anhydrous Silica. The pharmaceutically acceptable excipients are selected from lubricants such as Magnesium Stearate and moisture protective excipients such as hydroxy propyl methyl cellulose in combination with microcrystalline cellulose.

The granules of Hemidesmus indicus are prepared by blending root powder, dry starch, diluent, disintegrant and the blend obtained is dry granulated. The granules are coated with moisture protective excipients and lubricated with lubricants such as Magnesium Stearate and filled into hard gelatin capsules.

The granules may be further compressed with pharmaceutically acceptable excipients into tablets by conventional techniques and optionally coated with protective coating.

The herbal composition maybe in the form of powder filled into hard gelatin capsules or compressed into tablet.

Roots of Hemidesmus indicus and Hemidesmus Indicus var pubescens commonly known as "anantamool" by the locals were collected during three seasons i. e. summer, monsoon and winter from Mumbai, Melghat (Maharashtra state) and Trichur (Kerala state). Both the plants were studied for its physical investigation but only Hemidesmus indicus was studied in animals and in humans. Both the plants were authenticated from National Institute of Science Communication (NISCOM), New Delhi, India. They were assigned voucher number PID - 1821 and PID-1826 respectively. The plant material was thoroughly washed with water to remove soil particles, dust and other extraneous matter drained to remove excess of water by spreading over filter-papers for 6 hours in shade away from sunlight, was then placed in a preset oven at 45 ± 5 °C and was allowed to dry for 4 days. Immediately after drying, it was powdered using an electric mixer-grinder and sieved through a BSS mesh No. 85 sieve and was stored in commercially available airtight polyethylene containers labeled with details such as date, time and region of collection, weight of powder, which was used for research.

Quality control methods like proximate analysis, which included moisture content, total ash content were carried out, also phytochemical qualitative, quantitative and elemental analysis were carried out. Methanolic extract of plant, whose stability was checked was found more appropriate than other extracts like water etc. and was used to develop fingerprinting on High Performance Thin Layer Chromatographic instrument.

This extract may be converted into a pharmaceutically acceptable form and used in composition along with pharmaceutically acceptable excipients.

The plant material was subjected to preliminary phytochemical screening for detection of various plant constituents.

10 grams of air-dried powdered plant material was extracted successively with solvents like Petroleum ether (60-80), Benzene, Chloroform, Acetone, Ethanol (95%) in a soxhlet extractor.

Each time before extracting with next solvent, the powdered material was dried in oven below 50° C. Finally the marc was macerated with chloroform water for 24 hours to obtain the aqueous extract. Each extract was concentrated by distilling off the solvent and evaporating it to dryness on water bath. Extract thus obtained from each solvent was weighed; its percentage in terms of air-dried weight of plant material was calculated. The color and consistency of extract was also noted. The extracts obtained as above were then subjected to qualitative tests for the identification of various plant constituents such as alkaloids, carbohydrates, glycosides, phytosterols, oils, fats, saponins, phenolic compounds, tannins, proteins, free amino acids, gums, mucilages and volatile oils. The data of preliminary phytochemical constituents is presented in table 1. TABLE I PRELIMINARY PHYTOCHEMICAL ANALYSIS OF HEMIDESMUS INDICUS (H.INDICUS) AND HEMIDESMUS PUBESCENS (H.PUBESCENS)

Note: Present: -"+" and Absent: - "-"

All the experiment was repeated three times. Table II

Percentage of ethanol and water extractable matter in Hemidesmus indicus and pubescens

ASH CONTENT-The ash remaining following ignition of medicinal plant materials

(root) was determined by different methods, which measure

^■φ- Total ash

-Φ- Acid-insoluble ash

The total ash method was designed to measure 'physiological ash', and 'non- physiological ash'. Acid-insoluble ash is the residue obtained after boiling the total ash with dilute hydrochloric acid and igniting the remaining insoluble matter. Moisture content was determined by Karl Fischer titrimetric method. Crude fiber content was also estimated.

Following results of total ash, acid insoluble ash, moisture content and crude fiber content is given in table III:- Table III : Physical parameters of H Indicus

* Each observation is mean of three readings.

Elemental analysis was carried out with the known amount of root powder, and the percentage values of the elements are given in table IV. Table IV : Percentage of elements in Hemidesmus indicus and pubescens.

Also some quality control methods were developed for quick identification of both the plants. Plant powder was mixed with reagents like CaCl_2> NaOH etc and the visible change in the color were noted.

TABLE V : Reactions of reagents with Hemidesmus indicus and pubescens

These quality control parameters are depending on cultivation, handling of the raw material and storage conditions. The present invention also involves basic extraction procedures for crude root powder to obtain the therapeutically desirable portion and eliminate the unwanted material by treatment with a selective solvent, known as the menstruum. The principal methods of extraction are maceration, percolation, digestion, infusion and decoction. The quality of the finished product can be enhanced by optimization of primary extracts. The United State Pharmacopoeia (USP) provides general guidelines for maceration and percolation under the heading of tinctures. The solvents used for extraction were methanol, chloroform, petroleum ether (40 - 60°C) and water.

The root powder is standardized in a way to have the following phytochemical composition: alkaloids 6.5% to 7.5%, fatty acids 3.7 % to 4.2% , tannins 0.2% to 0.3%, phenolics 1.1 % to 1.2 % ,acidic flavonoids 5.2% to 5.4 % and basic flavonoids 0.7 % to 0.8%; elements such as calcium 23.3% to 25.5%; magnesium 7.9% to 9.16%;chloride 0.008% to 0.016%;sulphate trace amount and iron 1.1% to 1.3%; and total ash content 4.3% to 5.3%, acid insoluble ash content 13.3% to 15.1%, moisture content 8.7% to 9.1% and crude fiber content 15.1% to 19.5%. In the present work different solvents, from non-polar to polar, were used to optimize the extractive values of Hemidesmus indicus. The percentage of phytochemicals such as alkaloids, fatty acids, tannins and phenolics from the root powder extraction in all three seasons are given in table VI. Table VI : Percentage of extraction of phytochemicals of H.Indicus

Each observation is mean of three readings.

The percentage of flavonoids in the extract was estimated in all three seasons and is shown in table VII.

TABLE VII : Percentage of flavenoids in Hemidesmus indicus and pubescens

Optimization of amount of solvent and time of extraction for various solvents was carried out and the observations are shown in table VIII.

Table VIII Optimization of amount of solvent and time for extraction of Hemidesmus indicus in different solvents

• Each observation is mean of three readings. Water is the best solvent for extraction of Hemidesmus indicus as maximum extraction is achieved (28%) followed by methanol (15.4%). Also the time required in both cases is 2 hrs.

Optimization of number of extractions was also carried out and the observations are shown in table IX.

Table IX : Optimization of number of extractions for extraction of Hemidesmus indicus

These optimized parameters can be used for bulk production of whole plant powder extracts of Hemidesmus indicus.

The herbal composition comprises Hemidesmus indicus, diluents such as maize starch, disintegrant such as colloidal anhydrous silica, lubricant such as magnesium stearate and dry starch. The dosage form is capsule; particularly hard gelatin capsules. Further the invention is exemplified by non-limiting example. EXAMPLE Herbal Composition

The granules of Hemidesmus indicus are prepared by blending root powder, dry starch, Maize starch, Colloidal Anhydrous Silica and the blend obtained is dry granulated. The granules are coated with moisture protective excipients such as Hydroxy propyl methyl cellulose and microcrystalline cellulose and lubricated with lubricants such as Magnesium Stearate and filled into hard gelatin capsules. The above-mentioned formula was used for trials, and for animal study the powder was given as crude material. Accelerated stability studies as per US-FDA guidelines for stability were carried out. The samples were stored at room temperature and at 40°C ± 2°C; 75 % ± 5 % RH (relative humidity) for period of six months. Granules as well as capsules were tested for stability. Three months stability data is provided in the following table XL Storage conditions were below 30°C, 60% ± 5% RH Packing: Blister strip of 10 capsules Table XI Stability data of granules as well as capsules of Hemidesmus indicus

The herbal composition is administered orally. The subjects to be treated are preferably subjects in need of treatment (suffering from a kidney dysfunction). Subjects to be treated are preferably mammals, particularly humans. Kidney dysfunction to be treated includes primary glomerulonephritis (biopsy proven) or aminoglycoside induced nephrotoxicity, hence the herbal composition is useful in glomerular as well as tubular dysfunction.

Advantageously, the herbal compositions and methods of the present invention provide an alternative treatment option for subjects suffering from kidney dysfunction with high lipid level. In addition, the herbal composition and method of the present invention provide an alternative treatment option that exhibits no side effects. These and other advantages of the present invention will become readily apparent from the toxicity and efficacy studies set forth below.

Toxicity and' efficacy studies were carried out and the data is presented.

Acute toxicity studies on Hemidesmus indicus Linn is carried out on male and female Albino Swiss Mice weighing 35 to 55gms using Hemidesmus indicus sieved root powder. Three groups were made each comprising 3 males and 3 females. 2.0 ml of dose per animal was administered through Intragastric administration with help of Gavage No. 16 using distilled water as vehicle. A single administration of following dose concentrations was made 3.0, 5.0. and 7.0 g/kg body weight. Acclimatization was carried out for 14 days, one day drug administration and 14 days observation period. During the study the mice feed was supplied by AMRUT FEED.

Cage side observations - Usual cage side observations as expected in general toxicological studies were carried out and were found normal. No mortality was found in any of the animals even at the highest doses in both the sexes. In toxicity studies which were carried out in mice proved that test drug is not toxic even at dose of 7.0-g/kg-body weight., as no mortality was observed.

Efficacy studies on Hemidesmus indicus Linn: -

Efficacy studies on Hemidesmus indicus Linn is carried out as a potential nephroprotectant on a gentamicin induced kidney damaged rat model.

Animal models used for the efficacy study were male and female Albino Wister Rats weighing 110 to 150gms. The animals were randomly divided into five groups of ten animals (six males + six females),Group I normal Control, Group II gentamicin treated and control, Group III gentamicin treated and natural recovery and Group IV gentamicin + herbal composition treated.

Reversible kidney damage was induced by an intra-muscular (i.m) injection of gentamicin sulphate 90 mg kg^"1 in 1 cm³ distilled water to each animal from Group II - IV. A dose of 0.2334g/-kg root plant powder suspended in 2 cm³ distilled water was administered to each rat of Group IV. The normal control, gentamicin control and gentamicin natural recovery group animals were also administered 2 cm³ distilled water each. The mode of dosing was oral via a gavage. The animals were first given gentamicin injection intramuscularly then the oral dose of the herbal composition. The animals from Group I, III and IV were sacrificed at 6 days after gentamicin damage, and the ones from Group II were sacrificed on seventh day of the study. The dosage regime is tabulated in Table XII

TABLE -XII : Efficacy study of Hemidesmus indicus, daily dosage regimen

Note : 1. The above dosage is for an individual animal of the group. 2. The number of animals in each group = 12 (six male + six female). 3. i.m. = intra muscular.

Cage side observations included general behavioral changes, daily food and water consumption and daily body weight changes. Along with blood biochemistry parameters, urine examination, histopathological evaluations of the kidney under light were carried out.

Food was discontinued 18 hours prior to sacrifice for each group but water was provided ad libitum. Each animal's urine was collected and stored in marked glass vials and was sent immediately for analysis. Blood was withdrawn by cardiac puncture under ether anesthesia. The animals were sacrificed by overdose of ether. Weights of the animals were recorded, before sacrifice. Blood samples were collected in clean and pre- heparinised vials. Each vial was labeled properly with the identification number corresponding to the animal and the vials with blood were immediately stored at 4° C. The blood samples were later analyzed (within 8 hour after collection) for blood biochemical parameters like levels of creatinine, blood urea nitrogen, electrolytes, total proteins, albumin, cholesterol, triglycerides, AST, ALT, alkaline phosphatase using STAT FAX 2000 Autoanalyser. All enzyme assays were completed first, to avoid possible variations in enzyme levels due to storage.

Kidney was excised during autopsy, rinsed in 0.9 % saline and blotted dry of saline and excess of blood and weighed. A small piece (3mm x 3mm) of the central portion of the largest lobe of the kidney was cut and fixed in Bouin's fixative (Picric acid: Formalin: Acetic acid in the proportion 75: 25: 5) for histopathological examination.

No behavioral changes were observed. The food consumption of the animals from gentamicin control, gentamicin + herbal composition treated decreased but not significantly. The gentamicin recovery group animals showed decrease up to the sixth day of the treatment, which did not come to its predose level, but the decrease was not significant. The body weight showed decrease in body weights on the fifth day of the treatment. Gentamicin + herbal composition treated group animals also showed decrease in body weights. The natural recovery group animals showed increase in body weights after the tenth day of the treatment. Plant control group animals showed no significant variations in their body weight changes. The decrease in body weights which though not significant indicates toxicity due to gentamicin.

Liver to body weight ratio showed a decrease in the animals of Group - II, III and IV as compared to Group - 1 animals. Kidney to body weight ratio showed an increase in all the groups as compared to Group-I animals. Reduction in this ratio was evaluated in Group-IV, when compared with Group-Ill. Organ to body weight ratio of the rats from plant treated groups showed better recovery as compared to gentamicin natural recovery group.

Gentamicin treated rats in Group-II, III, IV developed hypercholesterolaemia and hyper-triglyceridaemia, with increase in serum creatinine, BUN level, γGT, bilirubin, alanine transamine, alkaline phosphatase. The progressive rise in BUN with gentamicin administration indicated a decrease in the functional capacity of kidney due to acute renal failure. Raised levels of creatinine substantiated these findings. After oral administration of root slurry the plasma levels of urea, creatinine, γGT, bilirubin, alanine transamine, alkaline phosphatase were found to be decreased in gentamicin treated plant Group. Levels of serum potassium, chloride, sodium, total protein, albumin were not different when the comparison was made between gentamicin groups and normal control group.

Decrease in hemoglobin in peripheral blood was seen in the animals from Group- II. No significant differences in red blood cell, white blood cell, neutrophil, monocyte or lymphocyte counts were seen between normal and gentamicin treated groups.

Controls remained free of Proteinuria, Occult blood, pus cells and red blood cells. Heavy proteinuria was observed in Group-II, III when compared with Group-IV (2.25±0.96, 2.33±0.58 vs l±O, gentamicin control, gentamicin natural recovery group vs gentamicin plant treated group of male rats at termination of study p<0.001, Table-X a and b). Rats from Group-II, III showed presence of Occult blood, pus cells and red blood cells in both male as well as female rats, with absence of above in Group- IV.

Effect of gentamicin and root powder of H. mdicus on organ to body weight ratio and blood biochemical parameters in male rats

Gentamycin Normal Gentamycin Gentamycin Test + Root Control Control Recovery slurry

Liver/ bd wt 0.03 0.026 0.029 0.029 ratio ±0.0054 +0.0023 ±0.0032 ±0.0026

Kidney/ 0.003 0.003 0.003 0.003 Rt bd wt ±0.0006 ±0.0002 ±0.0003 ±0.0002 ratio 0.003 0.003 0.0035 0.0034 Lt ±0.0006 ±0.0004 ±0.000 ±0.000

Haemoglobin 16.3 ±0.46 13.3 ±1.90 15.8 ±1.55 17.0 ±1.60

S. Creatinine 0.6 ± 0.13 0.73±0.10 1.0+0.16 0.9±0.10*

BUN 16.4±2.30 21.1+2.84 18.1+2.40 18+1.6*

117.1 TC 86.8±5.06 98.8±4.58 110±20.2 ±11.63

S.G.O.T 39.8±5.23 81+4.65 64.5±4.09 52.5±3.39

S.G.P.T 56.9±4.31 78.8±8.68 67.1+5.52 54.1±5.22 28.17 7.83 γGT 7.83±1.94 20±6.26 ±11.43 ±1.94** 73.33 147.83 A P 220±44.11 159+21.59 ±16.84 ±24.57

116.6 Triglyceride 86.8±5.06 191.5+6.59 138.8±4.83 ±9.02**

S. Billirubin 0.8±0.14 0.68+0.15 0.77±0.14 0.73±0.12

All the values are expressed as mean ± standard deviation, where n=6

Differences, which are significant, are marked as: * p< 0.01, ** p<0.002 and *** pθ.0001

TC-Total Cholesterol, AP-Alkaline phosphate

Effect of gentamicin and root powder of H. indicus on organ to body weight ratio and blood biochemical parameters in female rats

Gentamycin Normal Gentamycin Gentamycin Test + Root Control Control Recovery slurry

Liver/ bd wt 0.03 0.025 0.031 0.031 ratio ±0.0044 ±0.0021 ±0.0029 ±0.0029

Kidney/ 0.003 0.002 0.0042 0.003 Rt bd wt ±0.0004 ±0.0003 ±0.000 ±0.0001 ratio 0.003 0.002 0.004 0.003 Lt ±0.0004 ±0.003 ±0.0006 ±0.0002 Haemoglobin 14.2±0.70 12.8+1.21 16.6±0.82 17.1±0.74

S. Creatinine 0.7±0.16 0.7±0.26 0.97+0.11 0.8±0.2*

BUN 18.5+1.48 21.5+4.81 17.1+0.70 17+1.17*

TC 135.8+11.4 135+12.82 134.5±20.8 108.1+7.30

S.G.O.T 38.91±4.56 63±5.96 56±2.89 38.7±4.27

S.G.P.T 44.8±4.12 68±8.39 57.5+3.61 47.93±3.79

22.17 γGT 8.0±3.32 40.83+4.44 15+4.86** ±15.25

207.33 191.17 AP 70±34.88 180 ±38.07 ±11.34 ±30.15

110.6 203.3 110.6+ Triglyceride 204±7.24 ±12.13 ±15.32 12.13**

S.Billirubin 0.7±0.13 0.78±0.15 0.87±0.20 0.5±0.10

All the values are expressed as mean ± standard deviation, where n=6

Differences, which are significant, are marked as: * p< 0.01, ** p<0.002 and *** pO.OOOl

TC-Total Cholesterol, AP- Alkaline phosphate

Effect of gentamicin and root powder of H. indicus on urine composition

Gentamyci Gentamyci Gentamyci

Parameter Normal n n n Control Control Recovery Root slurry

Male 7.35±0.56 7.67 + 0.60 8.17 + 0.29 8 + 0.00 pH Female 7.23±0.45 6.33±0.82 8.17±0.29 4.37±3.28

Male Absent 2.25±0.96 2.33±0.58 l±O

Albumin Female Absent 2±1 1.67+1.15 1.5±0.70

Occult Male Absent 3.2±0.84 3.67±0.58 Absent

blood Female Absent 3±0.70 l±O 2±0

Male Absent 3±0 2.25±0.5 Absent

Pus cells Female Absent 2.25±0.5 Absent Absent

Male Absent 3±0 2.67±0.58 Absent

RBC's Female Absent 2.25±0.5 Trace 2±0

Casts/cyst Male Absent Absent Absent Absent s Female Absent Absent Absent Absent

All the above values are mean ± SD of 6 values.

SD- standard deviation

A dose of 90-mg/ kg body weight was effective to treat gentamicin induced nephrotoxicity( i.m for 6 days). Hemidesmus indicus prevented the fall in body weight. All the histopathological, biochemical changes showed Hemidesmus indicus preserves the functional capacity of kidney. The above picture of attenuation of damage was confirmed on histopathology.

In the present animal study, treatment with the herbal composition showed an increase in haemoglobin level. Reduction in the serum biochemical and enzyme levels, proteinuria and other parameters studied in herbal composition treated rats as compared to the gentamicin control and recovery rats suggests the potential in the use of the herbal composition in the treatment of gentamicin induced nephrotoxicity. Hence the treatment with herbal composition helps in management of renal impairment induced by gentamicin in rats.

The animals showed increase in body weight, also the food and water consumption was good. It was also found that urine samples were free of proteinuria, occult blood, pus cells and red blood cells. There was also reduction in organ/ body weight ratio (kidney/ body weight, liver/body weight ratio) in these animals. Histopathological investigations done on the kidney tissue of the animals in drug induced group revealed that the patterns of lesions in the tubules and glomeruli showed reduction, also at certain foci the changes were mild with less cellularity and less of swelling in the epithelium, when compared with other groups like gentamicin control, gentamicin natural recovery. Hence reduction in serum biochemical and enzymes levels, proteinuria and other parameters in the rats that were administered with herbal composition suggest the potential of the herbal composition as renoprotectant as well as lipid lowering agent. Clinical trial was carried out with Hemidesmus indicus .

A study to evaluate the efficacy and safety of Hemidesmus indicus in patients with chronic renal failure due to primary glomerulonephritis was conducted on 50 patients diagnosed with chronic renal failure [mild to moderate] secondary to glomerulonephritis. To acquire a true perspective, the trial was designed as a double blind randomized placebo-controlled trial. Patients of both sexes were included in the study. They were guided to take therapeutic doses of vitamins and minerals and antihypertensive medications or other medications if they were on for their disease and test drugs were administered as per dosage schedule mentioned in the Performa. The patients were advised to exclude certain food products like: curd, cold beverages, ice cream, and fermented food products till about 12 weeks during the treatment period.

Scoring was given to different symptoms like:

Polyuria Grade — 1, 2 (1 = day, 2 = nocturia)

Edema Grade — 1, 2, 3, 4 (1 = localized to one leg, 2 =generalized form (in periorbital areas), 3 = puffiness of face, 4 = generalized + localized)

Vomiting Grade — 1, 2 (1 = 1-3 daily, 2 = more than thrice)

Pallor (yes/no)

Breathlessness Grade — 1, 2, (1 = breathlessness on exertion, 2= breathlessness without exertion)

Hypertension > Grade — 1, 2,3 (1= mild, diastolic>90- 100mm hg), 2= moderate, diastolic> 100-110mm hg, 3=severe, diastoliOl 10mm hg)

Lethargy (yes/no)

Mental disturbances Grade — 1, 2, 3 (1 = lack of power of concentration, 2 = forgetfulness, 3 = irritability)

Anorexia (yes/no)

Investigations were carried out at baseline (0- Week), fourth (4^th week), eight (8th -

Week), and twelfth (12th - Week):

Follow up was carried on all the following weeks Total blood count, Reticulocyte count

(if necessary), LFT (Liver Function Test like SGOT, SGPT, GGT), S.calcium, S. triglycerides, S. cholesterol, BUN, S. creatine, S. sodium, Bilirubin, S. potassium, S. chloride, Urinalysis (protein quantitation), ECG, Chest x-ray all these investigations Tϊ-T TIJ were carried out on O week, 4 ,8 , 12 Week respectively.

The following chart gives the details of the tests, which were carried out:

Selected patients were randomly allocated to 2 groups A and B, to receive herbal composition dosage according to the batch corresponding to their codes, of one capsule four times daily for 12 weeks. The test herbal composition and the placebo were supplied in blister packs of 120 capsules which was sufficient dosage for 1 month of therapy. Selected patients were examined thoroughly clinically and investigated for Biochemical and metabolic parameters as per Performa. Inclusion Criteria was :-Age 18-60 yrs, Patients of either Sex, Cases of Chronic renal failure due to primary glomerulonephrites proved by kidney biopsy,GFR<50 ml/min (Calculated according to Cockroft and Gault)

Exclusion Criteria was:- Pregnant or lactating women, etiology of CRF other than primary Glomerulonephritis, like pyelonephritis, cystic disease, multisystemic disease (DM also SLE and Amyloid vascular disease), drug toxicity, hereditary, malignancy, Cardiac failure and cardiomegaly, peripheral neuropathy (initial sensory, drowsiness, convulsions, finally coma), those receiving steroids.

Adverse Drug reaction: Special look out for symptom's like nausea, vomiting, drug allergy, diarrhea, headache, hyperacidity, anorexia, dysuria, oligouria, constipation, sleep disturbances, breathlessness, cough, rhinitis, tachycardia, disturbances in blood pressure was kept..

All the data was analyzed statistically.

Calcium lactate was used as Placebo. Both the herbal composition and the placebo were filled in Capsules no-0 using hand filling capsule machine. All the capsules were kept in airtight plastic containers away from moisture and excessive heat (specification of capsules is given earlier).

All 10 patients completed 12 weeks of treatment. On decoding it was found that 5 patients (4 male and 1 female) received drug and 5 patients (4 male and 1 female) received placebo. Their average weight was 30.4 ± 10.4 and 37.4 ± 11.8 respectively. The mean age group in herbal composition treated group was 37 years (range, 21 to 52 years) while the mean age of patients receiving Calcium lactate (placebo) was 30 years (range, 20 to 42 years).

The presenting features in two groups included the following parameters as given in Table- XIII. Table- XIII : Investigations of general health parameters

Other parameters like ECG and chest X-ray did not show any difference after completion of the study. In addition to effects of herbal composition on various body parameters, tolerance of the herbal composition was also assessed. Along with biochemical parameters, clinical estimation was also done. Statistical analysis for non- parametric data e.g. Wilcoxon Matched-Pair test was also carried out to see if there was any significant change.

There was a decrease in S. creatinine levels from baseline(2.3 ±0.7) to (2.2±1.1), in BUN levels from baseline(40.4 ±24.5) to (33.2±16.1) , in S. triglycerides levels from baseline(119.2 ±61.2) to (115.4+28.4), in S. cholesterol levels from baseline( 193.8 ±61.24) to (163±51.2), in diastolic blood pressure from baseline (110 ± 5.4) to (94 ± 5.4) and in systolic blood pressure from baseline(134 ±9.6) to (128+13.0), after treatment for 12 weeks when compared to placebo which showed increase. The herbal composition did not change any vital body parameter like systolic or diastolic blood pressure, weight etc. In the patients on test herbal composition there was reduction in biochemical parameters like S. creatinine, BUN, S. triglycerides, S. cholesterol and also in Blood pressure with increase in hemoglobin levels with good tolerance level. There was no adverse effect on any investigative parameter. These investigations showed that the herbal composition did not produce any deleterious effect on kidney function. Hence it appears to be safe for use in cases of moderate renal dysfunctions.

Claims

I Claim

1. A Renoprotective and lipid lowering herbal oral composition comprising granules of Hemidesmus indicus along with pharmaceutically acceptable excipients.

2. The herbal composition as claimed in claim 1, wherein said granules of Hemidesmus indicus comprise standardized root powder, dry starch powder, diluent and disintegrant.

3. The herbal composition as claimed in claim 1, wherein said diluent used is Maize starch and said disintegrant used is Colloidal Anhydrous Silica.

4. The herbal composition as claimed in claim 1, wherein said granules comprises 250mg standardized root powder, 50mg Maize starch; and 3mg Colloidal Anhydrous Silica.

5. The herbal composition as claimed in claim 1, wherein said pharmaceutically acceptable excipients are selected from lubricants such as Magnesium Stearate and a moisture protective excipients such as Hydroxy porpyl methyl cellulose in combination with microcrystalline cellulose.

6. The herbal composition as claimed in claim 1, wherein, said standardized root powder comprises alkaloids 6.5% to 7.5%, fatty acids 3.7 % to 4.2% , tannins 0.2% to 0.3%, phenolics 1.1 % to 1.2 %, acidic flavonoids 5.2% to 5.4 % and basic flavonoids 0.7 % to 0.8%.

7. The herbal composition as claimed in claim 1, wherein elements of said standardized root powder comprises calcium 23.3% to 25.5%; magnesium 7.9% to 9.16%;chloride 0.008% to 0.016%;sulρhate trace amount and iron 1.1% to 1.3%.

8. The herbal composition as claimed in claim 1, wherein said standardized root powder comprises total ash content 4.3% to 5.3%, acid insoluble ash content 13.3% to 15.1%, moisture content 8.7% to 9.1% and crude fiber content 15.1% to 19.5%.

9. The herbal composition as claimed in any of the preceding claims, wherein said composition is in the form of powder or granules.

10. The herbal composition as claimed in any of the preceding claims, wherein said powder or granules are filled into hard gelatin capsule.

11. The herbal composition as claimed in any of the preceding claims, wherein said powder or granules may be compressed into tablets.

12. The herbal composition as claimed in any of the preceding claims, wherein said tablets may be coated or uncoated.

13. The herbal composition as claimed in any of the preceding claims, is used in the treatment of primary glomerulonephritis (biopsy proven) and/or aminoglycoside induced nephrotoxicity.

14. The herbal composition as claimed in any of the preceding claims, is used in the treatment of glomerular as well as tubular dysfunction.

15. Method of treatment comprising administering a therapeutically effective amount of said herbal composition to a subject in need suffering from primary glomerulonephritis (biopsy proven) and/or aminoglycoside induced nephrotoxicity.

16. The herbal composition as claimed in any of the preceding claims, wherein the administration of said herbal composition in warm blooded animals increases Hb level due to the presence of iron content.

17. The herbal composition as claimed in any of the preceding claims, wherein the administration of said herbal composition in warm blooded animals decreases serum triglyceride levels, serum creatinine, serum cholesterol, systolic and diastolic blood pressure.

18. The herbal composition as claimed in any of the preceding claims, wherein the administration of said herbal composition in warm blooded animals decreases in organ to body weight ratio particularly in case of kidney.

19. The herbal composition as claimed in any of the preceding claims, wherein the administration of said herbal composition in warm blooded animals increases in body weight of said animals and an intake of food and water.