U.S. patent application number 12/004403 was filed with the patent office on 2008-06-12 for hepatocurative effect of emblica officinalis against cyp 450 bio-activation hepatotoxicity of drugs.
Invention is credited to Sheikh Tasaduq Abdullah, Kasturi Lal Bedi, Devinder Kumar Gupta, Satinder Mohan Jain, Rakesh Kamal Johri, Bal Krishan Kapahi, Dilip Manikrao Mondhe, Ghulam Nabi Qazi, Kuldeep Singh, Om Parkash Suri.
Application Number | 20080138452 12/004403 |
Document ID | / |
Family ID | 28452466 |
Filed Date | 2008-06-12 |
United States Patent
Application |
20080138452 |
Kind Code |
A1 |
Johri; Rakesh Kamal ; et
al. |
June 12, 2008 |
Hepatocurative effect of emblica officinalis against CYP 450
bio-activation hepatotoxicity of drugs
Abstract
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
Inventors: |
Johri; Rakesh Kamal; (Jammu,
IN) ; Abdullah; Sheikh Tasaduq; (Jammu, IN) ;
Singh; Kuldeep; (Jammu, IN) ; Gupta; Devinder
Kumar; (Jammu, IN) ; Kapahi; Bal Krishan;
(Jammu, IN) ; Mondhe; Dilip Manikrao; (Jammu,
IN) ; Jain; Satinder Mohan; (Jammu, IN) ;
Suri; Om Parkash; (Jammu, IN) ; Bedi; Kasturi
Lal; (Jammu, IN) ; Qazi; Ghulam Nabi; (Jammu,
IN) |
Correspondence
Address: |
SMITH, GAMBRELL & RUSSELL
1130 CONNECTICUT AVENUE, N.W., SUITE 1130
WASHINGTON
DC
20036
US
|
Family ID: |
28452466 |
Appl. No.: |
12/004403 |
Filed: |
December 21, 2007 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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11271747 |
Nov 14, 2005 |
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12004403 |
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10106119 |
Mar 27, 2002 |
7001619 |
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11271747 |
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Current U.S.
Class: |
424/769 |
Current CPC
Class: |
A61P 43/00 20180101;
A61K 2300/00 20130101; A61K 36/47 20130101; A61K 36/47
20130101 |
Class at
Publication: |
424/769 |
International
Class: |
A61K 36/47 20060101
A61K036/47; A61P 43/00 20060101 A61P043/00 |
Claims
1. An orally-administered composition useful for treatment of 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, said composition comprising an substantially pure extract
from Emblica officinalis and pharmaceutically acceptable additives,
wherein the extract comprises 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, and 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.
2-4. (canceled)
5. A composition as claimed in claim 1, wherein said additives have
no effect on hepatocurative effectiveness of the said extract.
6. A composition as claimed in claim 1, wherein said extract is
prepared by incubating said Emblica officinalis with a polar
solvent at room temperature for about 15 to 25 hours so as to
obtain said extract, said extract comprising from 8.5% to 15% by
weight of tannin, and optionally adding pharmaceutically acceptable
additives, wherein the polar solvent consists of (i) water, (ii)
water and ethanol, or (iii) ethanol.
7. (canceled)
8. A composition as claimed in claim 1, wherein said composition is
in form of capsule, tablet, syrup, concentrate, powder, granule,
aerosol, and/or beads.
9. A method of preparing an orally-administered composition useful
for treatment of 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, said composition comprising an substantially
pure extract from Emblica officinalis and pharmaceutically
acceptable additives, wherein the substantially pure extract
comprises from 8.5% to 15% by weight of tannin, said substantially
pure 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, and 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, said method comprising incubating said Emblica officinalis
with a polar solvent at room temperature for about 15 to 25 hours
so as to obtain the substantially pure extract, the substantially
pure extract comprising from 8.5% to 15% by weight of tannin, and
optionally adding pharmaceutically acceptable additives, wherein
the polar solvent consists of (i) water, (ii) water and ethanol or
(iii) ethanol.
10. (canceled)
11. A method as claimed in claim 9, wherein extract is from fresh
and/or semi dried fruits of Emblica Officinalis.
12-13. (canceled)
14. A method as claimed in claim 9, wherein said composition is in
form of capsule, tablet, syrup, concentrate, powder, granule,
aerosol, and/or beads.
15-38. (canceled)
39. A method for treating CYP 450 bio-activation hepatotoxicity
induced by drugs comprising treating a mammalian with a
hepatocurative effective amount of substantially pure extract from
Emblica officinalis and optionally pharmaceutically acceptable
additives.
Description
FIELD OF THE PRESENT INVENTION
[0001] 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.
BACKGROUND OF THE PRESENT INVENTION
[0002] Liver disorders are still the major health hazards both in
urban and rural areas of the world. Despite scientific advances in
our understanding of hepatotoxicity, and leads provided by
traditional system of medicine, we do not have yet any effective
entities to cure liver derangement more importantly those which are
caused by a variety of drugs. The Indian Council of Medical
Research, New Delhi in its revised research programme on
traditional medicines, has adopted liver diseases as one among six
thrust areas for multidisciplinary study.
[0003] The disorders of the liver may be classified into acute or
chronic hepatitis (inflammatory liver diseases), hepatosis
(non-inflammatory disorders). The acute condition is often followed
by liver cirrhosis as well as hepatic coma with grave prognosis.
Liver cirrhosis as such accounts amongst the ten top fatal diseases
in the world. Exposure of humans to a variety of agents such as
chemicals and drugs (xenobiotics), many natural compounds, viral
and bacterial pathogens with attendant predisposable conditions,
etc. are considered responsible for hepatic insufficiency.
[0004] There are large group of drugs which on repeated
administration produce liver toxicity. These are mediated primarily
by bioactivation so that the products of parent drug are toxic.
Another class of drugs induce toxicity by causing membrane rupture
or DNA damage and by interfering with protein synthesis. One of the
important categories of drugs are the anti-TB drugs which when
taken regularly cause hepatotoxicity. Tuberculosis is prevalent in
all countries of the world--tropical, subtropical and colder
regions. The chemotherapy of tuberculosis is important and
challenging, because the disease is often chronic and the toxicity
due to anti-TB drugs pose therapeutic problems. The disorders of
the liver caused during the treatment of tuberculosis, by known
antimicrobial agents range from jaundice to the fibrosis of the
liver. Several cases progress to the chronic form of disease or
have a fulminant course and prove fatal. Three drugs i.e.,
rifampicin, pyrazinamide and isoniazid comprise first choice
treatment of tuberculosis. These are to be administered for long
period of time and produce liver dysfunction leading to
toxicity.
[0005] Jaundice is amongst the most prominent incidence of their
adverse reactions. The characteristic pathology is the bridging and
multilobular necrosis. Hypersensitivity to these drugs leads to
hepatitis. Multidrug treatment also poses special problems.
Rifampicin causes liver damage. Disturbances in liver function is
more if it is combined with isoniazid. Pyrazinamide is the most
toxic of the three anti-TB drugs. Continuation of the drugs in
combination after symptoms of hepatic dysfunction have appeared
tends to increase further the severity of damage. Severe hepatic
injury leading to death has been reported in patients receiving
these drugs (Slivka, IL, Farmakol Toksikol-1989: 52; 82-85)
[0006] In our traditional system of medicines (Ayurveda), use of
several medicinal plants have been prescribed for alleviating liver
disorders. There are nearly forty indigenous polyherbal
formulations from more than 100 plants enjoying reputation of being
hepatoprotectives. However, none have been specified as a
therapeutic agent, which is able to protect the liver from injury
due to treatment of anti-TB drugs. This owes partly to the fact
that reports are scanty with regard to evaluation of plants/plant
products, which would seem focussed specially against hepatic
injury caused by drugs which produce toxicity as a result of
bioactivation.
[0007] There is thus a growing interest in the development of
herbal entities considered relatively safe for alleviating liver
disorders specifically caused by the anti-TB drugs.
[0008] Emblica officinalis Gaertn. (Hindi:Amla) (Euphorbiaceae) is
widespread in India, Ceylon, Malaya and China. The tree is common
in mixed deciduous forests of India ascending to 4500 ft on the
hills, cultivated in gardens and homeyards. It is a small or medium
sized deciduous tree, fruits depressed globose, 1/2 to 1 inch in
diameter, fleshy, and contains six trigonows seeds. The fruit is
sour and is occasionally eaten raw. The fruit pulp contains (%);
moisture 81.2, protein 0.5, fat 0.1, mineral matter 0.7, Ca 0.005,
Phosphorus 0.02 and Iron 1.2 mg/100 gm, nicotinic acid 0.2 mg (100
gm, vitamin C 600 mg/100 mg. (Medicinal plants of India, Satyavati
et al (ed.), ICMR, new Delhi, 1976, p 377). The potent vitamin
C-like activity has been located in the low molecular weight
hydrolysable tannins. Four such compounds emblicanin-A,
emblicanin-B, punigluconin and pedunculagin have been isolated from
the fresh pericarp. The first two compounds are naturally occurring
galloellagi-tannins (Ghosal, et al, IndJChem, 1996:353:941-948;
Bhattacharya et al, Phytomedicine, 2000: 7: 173-175)
[0009] The fruit is acrid, cooling, and diuretic. Dried fruit is
useful in haemorrhage, diarrhoea and dysentery. It has been
extensively use in anemia, liver diseases and dyspepsea. A
fermented liquor prepared from the fruits is used in jaundice.
Fruits are a reputed Ayurvedic rasayan (revitaliser, biological
response modifier) (Sharma P. V. Dravyaguna vijnana, Chaukhamba
Sanskrit Sansthan, Varanasi, 1978). Several pharmacological
properties are also reported. Leaf extracts have been found to be
anti-inflammatory (Summanen et al, Planta Medica, 1993:59: 666),
antioxidant (Jose and Kuttan, Clin. Biochem. Nutr, 1995:19:63-70),
hypolipidemic (Mathw:eta\, JEthnopharmacol, 1996:50:61-68), cell
growth inhibition (Psatima et al, ACS Symp. Ser, 1998, p 701).
Hepatoprotective activity of Emblica officinalis extracts against a
chemical viz., carbon tetrachoride induced liver toxicity has been
demonstrated (Jose J K & Kutten R, J. Ethnopharmacology
2000:72; 135-40; Bhattacharya et al, Phytomerdicine
2000:7:173-5).
[0010] The article by Sharma et al (Hum Exp. toxicol 2000:19;
337-84) suggests that Emblica Officinalis prevents genotoxicity
induced by benzopyrines. Benzopyrene is one of the prominent
environmental carcinogen which is a specific substrate for CYP 450
1A1. Both are clastogenic. However, the liver toxicity produced by
the drugs including anti-TB drugs is dependent on a great measure
to their bio-activation through multiple CYP isoforms, the most
prominent being CYP450 3A4. There are several agents, which reduce
CYP levels or reverse the micronuclei formation but are not
hepatoprotective. To cite an example, applicants have developed a
molecule, piperine which is a specific inhibitor of CYP 450 1A1,
but is not hepatoprotective. Similarly, there are several known
compounds, which reverse the genotoxicity but are not
hepatoprotective. Therefore the decrease in CYP 1A1 or reversal of
clastrogencity can not be construed as hepatoprotection. For
example, jaundice (hepato-biliary dysfunction) has not been
correlated with genotoxicity, rather it may be an early event in
the onset of liver toxicity. In the present article, a casual
relationship between CYP decrease or genotoxicity has not been
related to attenuation of clinical pathology usually seen in
symptoms of hepatotoxicity.
[0011] In the present invention, the Applicants provide protection
against hepatotoxicity produced by all such 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.
[0012] Thus, we claim preparations from Emblica officinalis which
are superior so far as their systemic effects are manifested in
clinical profile (serum/liver parameters) which correlate to their
hepatoprotective profile.
OBJECT OF THE PRESENT INVENTION
[0013] The main object of the present invention is to develop a
hepatocurative composition against CYP 450 bio-activation
hepatotoxicity induced by drugs.
[0014] Another main object of the present invention is to develop a
hepatocurative composition against CYP 450 bio-activation
hepatotoxicity induced by anti-TB drugs.
[0015] Yet another object of the present invention is to develop a
composition from fruit Emblica officinalis for hepatocurative
effect.
[0016] Still another object of the present invention is to develop
a composition from fruit Emblica officinalis for hepatocurative
effect against CYP 450 bio-activation hepatotoxicity.
[0017] Still another object of the present invention is to develop
a method of preparing an extract from fruit Emblica
officinalis.
[0018] Still another object of the present invention is to develop
a method for treating a subject for CYP 450 and free radical
mediated hepatotoxicity caused by drugs using composition
comprising an extract from Emblica officinalis.
[0019] Still another object of the present invention is to develop
a method of using hepatocytes to understand the effect of extract
from fruit Emblica officinalis.
SUMMARY OF THE INVENTION
[0020] 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
[0021] 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.
[0022] 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.
[0023] 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.
[0024] In yet another embodiment of the present invention, wherein
said composition is administered orally.
[0025] In still another embodiment of the present invention,
wherein said extract and additives are in the ratio ranging between
1:1 to 1:10.
[0026] In still another embodiment of the present invention,
wherein said additives have no effect on the hepatocurative effect
of the said extract.
[0027] 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.
[0028] In still another embodiment of the present invention,
wherein said composition shows tanin content in the range of 8.5 to
15%.
[0029] 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.
[0030] 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.
[0031] In another embodiment of the present invention, wherein said
fruit is incubated with polar solvent at room temperature for about
15-25 hours.
[0032] In yet another embodiment of the present invention, wherein
said extract is from fresh and/or semi dried fruits of Emblica
Officinalis.
[0033] In still another embodiment of the present invention,
wherein said extract and additives are in the ratio ranging between
1:1 to 1:10.
[0034] 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.
[0035] 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.
[0036] 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.
[0037] In another embodiment of the present invention, introducing
drug toxicity in hepatocytes.
[0038] In yet another embodiment of the present invention, adding
said composition to said hepatocytes exposed to drug
hepatotoxicity.
[0039] 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.
[0040] In still another embodiment of the present invention,
wherein said method is particularly effective against
hepatotoxicity caused by anti-TB drugs.
[0041] 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.
[0042] In still another embodiment of the present invention,
wherein said composition is administered orally.
[0043] 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.
[0044] In still another embodiment of the present invention,
wherein said composition is useful for treating animals or human
beings.
[0045] In still another embodiment of the present invention,
wherein said composition has no adverse effect on health.
[0046] In still another embodiment of the present invention,
wherein said drugs are selected from a group comprising
Paracetamol, CCl.sub.4, and anti-TB drugs.
[0047] In still another embodiment of the present invention,
wherein said anti-TB drugs are selected from a group comprising
Rifampicin, Pyrazinamide, and isoniazid.
[0048] 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.
[0049] In still another embodiment of the present invention,
wherein said method uses hepatocyte culture for ideal insight.
[0050] 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.
[0051] In still another embodiment of the present invention,
wherein said composition is useful for treating animals or human
beings.
[0052] In still another embodiment of the present invention,
wherein said composition has no adverse effect on health.
[0053] In still another embodiment of the present invention,
wherein said composition shows restoration of hepatocyte
viability.
[0054] In still another embodiment of the present invention,
wherein said method shows prevention of cell membrane leakage.
[0055] 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.
[0056] In still another embodiment of the present invention,
wherein said composition reverses the leakage of glutamate pyruvate
transaminase (GPT) from hepatocyte.
[0057] 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.
[0058] 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.
[0059] 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.
[0060] In still another embodiment of the present invention,
wherein said method helps restore liver function to normal.
[0061] In still another embodiment of the present invention,
wherein dosage of said composition is ranging between 50-250
mg/kg.
[0062] 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.
[0063] 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.
[0064] 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.
[0065] Thus, applicants claim preparations from Emblica officinalis
which are superior so far as their systemic effects are manifested
in clinical profile (serum/liver parameters) which correlate to
their hepatoprotective profile.
[0066] 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 defense mechanism of the tissue, improve recovery from
dysfunctional states of the liver after prolonged challenge of
anti-TB drugs.
[0067] 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 fish 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.
[0068] 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.
[0069] 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.
[0070] 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.
[0071] 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.
[0072] 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).
[0073] 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).
[0074] 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.
[0075] 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
manifestations. 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.
BRIEF DESCRIPTION OF THE ACCOMPANYING DRAWINGS
[0076] FIG. 1 shows liver toxicity wherein plant extract attenuates
Rifampicin induced hepatotoxicity by restoring liver function to
normal (95% effect). The cell toxicity indicators shown in the said
FIG. 1 is the leakage of lactate dehydrogenase (LDH) from intact
cells after toxin challenge and its reversal by extracts.
[0077] FIG. 2 shows leakage of glutamate pyruvate transaminase
wherein said plant extracts in combination attenuates
Rifampicin+isoniazid induced hepatotoxicity by restoring liver
function to normal (96%). The cell toxicity indicators shown in
FIG. 2 is the leakage of glutamate pyruvate transaminase (GPT from
intact cells after toxin challenge and its reversal by the extracts
in combination. (Model: primary monolayer cultures of liver
cells).
[0078] FIG. 3 shows leakage of serum glutamate pyruvate
transaminase (GPT) after toxin challenge and its reversal by
Emblica officinalis fraction wherein Emblica officinalis reverses
Rifampicin+isoniazid+pyrazinamide induced hepatotoxicity and
restores the liver function to normal (96%). The cell toxicity
indicators shown in FIG. 3 is the leakage of serum glutamate
pyruvate transaminase (GPT) after toxin challenge and its reversal
by Emblica officinalis fraction.
[0079] FIG. 4 shows protection against cell leakage wherein,
Emblica officinalis fractions in combination prevents
Rifampicin+isoniazid induced toxicity. FIG. 4 shows that fractions
provide 96% protection against cell leakage as measured by serum
GPT levels and increases cellular defense. 75% increase in
glutathione levels (liver) is observed.
[0080] FIG. 5 shows Emblica officinalis extract reverses
Rifampicin+isoniazid+pyrazinamide induced toxicity. FIG. 5 shows
94% protection against rise in lipid Peroxidation (LPO, liver) and
96% decrease of serum Bilirubin in response to treatment with
extract of the present invention.
[0081] FIG. 6 shows a flow chart for the preparation of extract
from fruit Emblica Officinalis.
EXAMPLES
[0082] The invention of instant Application is further illustrated
by the following examples, which should not, however be construed
to limit the scope of the invention. The following examples are not
intended to be limiting in any way, but demonstrate some of the
preferred embodiments of the present invention. Any person skilled
in the art can design more combinations useful against drug-induced
toxicity, which may be considered as part of the present
invention.
Example 1
[0083] Table 1 shows that the plant products are effective against
paracetamol hepatotoxicity which is mainly dependent against
bioactivation mechanisms mediated by CYP 450. % protection is shown
as combined effect release of LDH and GPT in serum.
TABLE-US-00001 TABLE 1 Effect of plant products against
hepatotoxicity produced by paracetamol Treatment paracetamol
Toxicity 93% % protection as measured by combined effect against
LDH and GPT leakage Extract 97% protection Fraction 96%
protection
Example 2
[0084] Table 2 shows that the preparations of the present invention
are not effective against liver toxicity produced by agents where
the toxicity is primarily not dependent on bioactivation by CYP
450.
TABLE-US-00002 TABLE 2 Effect of plant products against
hepatotoxicity produced by galactosamine. Treatment galactosamine
Toxicity 93% % protection as measured by combined effect against
LDH and GPT leakage Extract 3% Fraction 7%
Example 3
[0085] Plant extract attenuates Rifampicin induced hepatotoxicity
by restoring liver function to normal (95% effect). The cell
toxicity indicators shown in FIG. 1 is the leakage of lactate
dehydrogenase (LDH) from intact cells after toxin challenge and its
reversal by extracts. (Model; primary monolayer cultures of liver
cells)
Example 4
[0086] Plant extracts in combination attenuates
Rifampicin+isoniazid induced hepatotoxicity by restoring liver
function to normal (96%). The cell toxicity indicators shown in
FIG. 2 is the leakage of glutamate pyruvate transaminase (GPT) from
intact cells after toxin challenge and its reversal by the extracts
in combination. (Model: primary monolayer cultures of liver
cells).
Example 5
[0087] Emblica officinalis reverses
Rifampicin+isoniazid+pyrazinamide induced hepatotoxicity and
restores the liver function to normal (96%). The cell toxicity
indicators shown in FIG. 3 is the leakage of serum glutamate
pyruvate transaminase (GPT) after toxin challenge and its reversal
by Emblica officinalis fraction.
Example 6
[0088] Emblica officinalis fractions in combination prevents
Rifampicin+isoniazid induced toxicity. FIG. 4 shows that fractions
provide 96% protection against cell leakage as measured by serum
GPT levels and increases cellular defense. 75% increase in
glutathione levels (liver) is observed.
Example 7
[0089] Emblica officinalis extract reverses
Rifampicin+isoniazid+pyrazinamide induced toxicity. FIG. 5 shows
94% protection against rise in lipid Peroxidation (LPO, liver) and
96% decrease of serum Bilirubin in response to treatment with
extract of the present invention.
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