The present invention relates to a composition useful for hepatocurative effect against CYP 450 bio-activation hepatotoxicity induced by drugs, said composition comprising an extract from Emblica officinalis and optionally pharmaceutically acceptable additives and method of treating drug induced hepatotoxicity.

SUMMARY OF THE INVENTION

The present invention relates to a composition useful for hepatocurative effect against CYP 450 bio-activation hepatotoxicity induced by drugs, said composition comprising an extract from Emblica officinalis and optionally pharmaceutically acceptable additives and method of treating drug induced hepatotoxicity.

DETAILED DESCRIPTION OF THE PRESENT INVENTION

Accordingly, the present invention relates to a composition useful for hepatocurative effect against CYP 450 bio-activation hepatotoxicity induced by drugs, said composition comprising an extract from Emblica officinalis and optionally pharmaceutically acceptable additives and method of treating drug induced hepatotoxicity.

In one embodiment of the present invention, a composition useful for hepatocurative effect against CYP 450 bio-activation hepatotoxicity induced by drugs, said composition comprising an extract from Emblica officinalis and optionally pharmaceutically acceptable additives.

In another embodiment of the present invention, wherein said additives are selected from a group of nutrients comprising proteins, carbohydrates, sugar, talc, magnesium stearate, cellulose, calcium carbonate, starch-gelatin paste, and/or pharmaceutically acceptable carrier, excipient, diluent, or solvent.

In yet another embodiment of the present invention, wherein said composition is administered orally.

In still another embodiment of the present invention, wherein said extract and additives are in the ratio ranging between 1:1 to 1:10.

In still another embodiment of the present invention, wherein said additives have no effect on the hepatocurative effect of the said extract.

In still another embodiment of the present invention, wherein said extract is prepared in a solvent selected from a group comprising aqueous, aqueous-ethanolic, ethanolic, ketonic, ethereal, halogenated solvents.

In still another embodiment of the present invention, wherein said composition shows tanin content in the range of 8.5 to 15%.

In still another embodiment of the present invention, wherein, said composition for the oral route is in form of capsule, tablet, syrup, concentrate, powder, granule, aerosol, and/or beads.

In further another embodiment of the present invention, a method of preparing composition comprising an extract from Emblica officinalis and optionally pharmaceutically acceptable additives, said method comprising steps of adding polar solvent to the fruit Emblica Officinalis to obtain the extract and optionally adding pharmaceutically acceptable additives.

In another embodiment of the present invention, wherein said fruit is incubated with polar solvent at room temperature for about 15-25 hours.

In yet another embodiment of the present invention, wherein said extract is from fresh and/or semi dried fruits of Emblica Officinalis.

In still another embodiment of the present invention, wherein said extract and additives are in the ratio ranging between 1:1 to 1:10.

In still another embodiment of the present invention, wherein said extract is prepared in a solvent selected from a group comprising aqueous, aqueous-ethanolic, ethanolic, ketonic, ethereal, halogenated solvents.

In still another embodiment of the present invention, wherein, said composition for the oral route is in form of capsule, tablet, syrup, concentrate, powder, granule, aerosol, and/or beads.

In further embodiment of the present invention, a method of treating a subject for CYP 450 and free radical mediated hepatotoxicity caused by drugs using composition comprising an extract from Emblica officinalis and optionally pharmaceutically acceptable additives.

In another embodiment of the present invention, introducing drug toxicity in hepatocytes.

In yet another embodiment of the present invention, adding said composition to said hepatocytes exposed to drug hepatotoxicity.

In still another embodiment of the present invention, measuring changes in the level of liver/serum markers to estimate hepatocurative effect of the said composition.

In still another embodiment of the present invention, wherein said method is particularly effective against hepatotoxicity caused by anti-TB drugs.

In still another embodiment of the present invention, wherein said composition is not effective against hepatotoxicity which is independent of bio-activation by CYP 450.

In still another embodiment of the present invention, wherein said composition is administered orally.

In still another embodiment of the present invention, wherein, said composition for the oral route is in form of capsule, tablet, syrup, concentrate, powder, granule, aerosol, and/or beads.

In still another embodiment of the present invention, wherein said composition is useful for treating animals or human beings.

In still another embodiment of the present invention, wherein said composition has no adverse effect on health.

In still another embodiment of the present invention, wherein said drugs are selected from a group comprising Paracetamol, CCl₄, and anti-TB drugs.

In still another embodiment of the present invention, wherein said anti-TB drugs are selected from a group comprising Rifampicin, Pyrazinamide, and isoniazid.

In still another embodiment of the present invention, wherein said composition controls abnormal rise in the clinical pathological symptoms revealed by serum/liver markers serving as indices of hepatic damage besides control of high levels of Bilirubin.

In still another embodiment of the present invention, wherein said method uses hepatocyte culture for ideal insight.

In still another embodiment of the present invention, wherein said drugs is used at cytotoxic levels to produce valid and reproducible results in liver cells.

In still another embodiment of the present invention, wherein said composition is useful for treating animals or human beings.

In still another embodiment of the present invention, wherein said composition has no adverse effect on health.

In still another embodiment of the present invention, wherein said composition shows restoration of hepatocyte viability.

In still another embodiment of the present invention, wherein said method shows prevention of cell membrane leakage.

In still another embodiment of the present invention, wherein said composition shows about 96% hepatocurative effect against combined effect of anti-TB drugs of Rifampicin, and Isoniazid.

In still another embodiment of the present invention, wherein said composition reverses the leakage of glutamate pyruvate transaminase (GPT) from hepatocyte.

In still another embodiment of the present invention, wherein said composition shows hepatocurative effect of about 96% against combined effect of anti-TB drugs of Rifampicin, isoniazid, and pyrazinamide.

In still another embodiment of the present invention, wherein said composition shows about 94% hepatocurative effect against rise in lipid Peroxidation (LPO) induced by combination of anti-TB drugs Rifampicin, isoniazid, and pyrazinamide.

In still another embodiment of the present invention, wherein said composition shows about 96% decrease of serum Bilirubin as a hepatocurative effect against combination of anti-TB drugs Rifampicin, isoniazid, and pyrazinamide.

In still another embodiment of the present invention, wherein said method helps restore liver function to normal.

In still another embodiment of the present invention, wherein dosage of said composition is ranging between 50-250 mg/kg.

In still another embodiment of the present invention, wherein said method is used for hepatocurative effect against drugs causing liver dysfunction, including anti-TB drugs.

In further embodiment of the present invention, the applicants provide protection against hepatotoxicity produced by all drugs, which are bio-activated by multiple CYP isoforms as indicated by clinical parameters in serum/liver. Besides, we claim that the clinical parameters showing toxicity are reversed even if the symptoms of genotoxicity may not begin to appear. For example decrease in abnormal rise of serum Bilirubin, which may be attributed to a protective effect also due to other cellular factors such as membrane stabilization, as revealed in primary monolayer cultures of liver cells.

Further, applicants have made use of their expertise and years to research to establish that Emblica Officinalis (Alma) cures hepatotoxicity induced by drugs that is restricted to CYP 450 bio-activation hepatotoxicity.

Thus, applicants claim preparations from Emblica officinalis which are superior so far as their systemic effects are manifested in clinical profile (serun/liver parameters) which correlate to their hepatoprotective profile.

In further embodiment of the present invention, it relates to preparations and methods of preparation and use of such products which restores the normal liver function against drug induced toxicity as a result of bio-activation of drugs applicable with particular relevance to anti-TB drug(s) induced liver toxicity. The compositions and methods of the present invention increase biological defence mechanism of the tissue, improve recovery from dysfunctional states of the liver after prolonged challenge of anti-TB drugs.

In another embodiment of the present invention, the compositions and methods of the present invention contain one of the extracts/fractions of Emblica officinalis fruit as an essential ingredient. These extracts/fractions may be obtained from fresh or semi dried fruits of Emblica officinalis. The compositions are formulated with more than one extracts and combined in any weight ratios. The preferred weight ratios include 1:1,1:2,1:1:1:, 1:2:2.2:1:2:, 2:2:1.

In still another embodiment of the present invention, it is related to preparation and use of products from Emblica officinalis, which restores normal liver function against drug induced toxicity caused as a result of bio-activation of drugs applicable with particular relevance to anti-TB drug(s) induced liver toxicity. The products of the invention comprise aqueous, aqueous-ethanolic, ethanolic, ketonic, ethereal, halogenated solvents extracts/fractions from Emblica officinalis, obtained either from fresh or semi-dried fruits. These contain 8.5-15% of tannin content. It also relates to preparation of compositions of such products in different proportions of more than one ingredient. These products either alone or in combination are intended to be used against drug induced liver toxicity as represented by specific mechanism underlying liver disorders and usually manifested in clinical conditions of liver dysfunction.

In still another embodiment of the present invention, the preparations either alone or in composition are also intended to be used against anti-TB drug(s).The use of such products as developed in the present invention controls the abnormal rise in clinical pathological symptoms revealed by serum/liver markers serving as indices of hepatic damage besides control of high levels of Bilirubin.

In still another embodiment of the present invention, in the development of the present invention the ample information has been utilized which exists regarding the advances in our understanding of mechanisms responsible for hepatotoxic injury due to drugs. More importantly the choice of a suitable model for the evaluation of anti-toxic profile of any substance is also crucial.

In still another embodiment of the present invention, a large body of information has been gained in favor of the present invention by using liver cell (hepatocyte) cultures, which ideally provide an insight into the mechanism of a toxin-induced impairment of hepato-biliary dysfunction because this model allows use of a test compounds (such as anti-TB drugs) to be used at cytotoxic level so that a valid and reproducible toxicity is generated. The in vitro cell culture model is of significant interest in ascertaining the mechanisms of toxicity and its reversal by protective agents.

In still another embodiment of the present invention, liver cells are considered as system of choice which have found ample application in the evaluation of cyto- cum geno-toxicity of chemicals and drugs (Nakagawa and Tayama, Arch Toxicol, 1995:69:208) and as such have been used in the evaluation of hepatoprotective profiles of the present invention. The mechanisms are revealed in critical biochemical functions of liver cells which are sensitive indicators of drug (s) toxicity. (Tomasi et al, Toxicol/Vtf/zo/:15:178-183). Both cellular lysis (measured by leakage of transaminases enzymes and lactated dehydrogenase from the cells) and the metabolic competence of the tissue are modified as a function of both the duration and concentration of the drugs (Goethals et al Fundm Appl Toxicol 1984:4:441-450).

In still another embodiment of the present invention, the preparations (alone or in combination) act in a specific manner. These act against toxicity produced by drugs including anti-TB drugs which require bioactivation by hepatic cytochrome P 450 dependent mixed function oxidases. Cytochrome P450 have been shown to be involved in the liver toxicity (Anundi I, Lindros K O, Pharmacol Toxicol 1992; 70;453-458). Participation of CYP 450 dependent oxidation of drugs including anti-tubercular drugs rifampicin, isoniazid, pyrazinamide in liver is reported (Ono et al, Biol Pharm Bull 1998:21:421-425).

In still another embodiment of the present invention, that the preparations mentioned in the present invention act in a specific manner is further evidenced by the observation that these preparations have not been found effective against galactosamine induced toxicity which is not dependent upon the intervention of mixed function oxidases. Of serious concern is the toxicity produced by use of some drugs in combination such as anti-tubercular drugs. Preparations alone or in combination, prevent not only (a) rifampicin (b) isoniazid (c) pyrazinamide induced toxicity but also various combinations of these such as (a) rifampicin+isoniazid (b) rifampicin+pyrazinamide (c) isoniazid+pyrazinamide and (d) rifampicin+isoniazid+pyrazinamide toxicity. The metabolic activation of drugs including anti-TB drugs alone or in combination is also accompanied by reactive intermediates which may be free radical/active metabolites/free oxy radicals through a variety of cellular oxidative metabolic pathways. An altered oxidative/antioxidative profile is closely associated with production of drug (s) induced hepatic injury (Sodhi et al, Hum Exp Toxicol 1997; 16;315-321). The efficacy of the products of the present invention is further shown by their anti-lipoperoxidant (anti-oxidant) profile. By using the preparations of the present invention a decrease in the accumulation of excess levels of the product of oxygen metabolism has been revealed.

In still another embodiment of the present invention, also included are the preventing role of the products developed herein, against cell membrane leakage and restoration of cell viability in challenged liver tissues caused as a result of toxic menifestations. Preparations act in a specific manner in as much as they prevent toxicity produced by bioactivation of drug (s) and combination of drugs (s) as described in the above art. The products described have no cytotoxicity and on the other hand enhance overall biological defense systems per se.
 

Claim 1 of 12 Claims

1. A method of treating CYP 450 and free radical mediated hepatotoxicity caused by at least one anti-TB drug selected from the group consisting of rifampicin, pyrazinamide, and isoniazid so as to restore liver function to normal, comprising:

(a) providing an orally-administered composition comprising an extract from Emblica officinalis and pharmaceutically acceptable additives, said extract comprising from 8.5% to 15% by weight of tannin, said extract and said additives being in the ratio of 1:1-1:10, the additives being selected from a group of nutrients comprising proteins, carbohydrates, sugar, talc, magnesium stearate, cellulose, calcium carbonate, starch-gelatin paste, and/or pharmaceutically acceptable carrier, excipient, diluent, or solvent, wherein said composition treats CYP 450 bio-activated hepatotoxicity induced by at least one anti-TB drug selected from the group consisting of rifampicin, pyrazinamide, and isoniazid so as to restore liver function to normal, but is not effective against hepatotoxicity which is independent of bio-activation by CYP 450

(b) introducing drug toxicity in hepatocytes of said subject,

(c) orally administering a hepatocurative dosage of said composition to a subject having said hepatocytes exposed to drug hepatotoxicity, and

(d) measuring changes in the level of liver/serum markers to estimate hepatocurative effect of the said composition.

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