U.S. patent application number 11/269594 was filed with the patent office on 2006-03-16 for composition comprising pharmaceutical/nutraceutical agent and a bio-enhancer obtained from glycyrrhiza glabra.
This patent application is currently assigned to COUNCIL OF SCIENTIFIC AND INDUSTRIAL RESEARCH, COUNCIL OF SCIENTIFIC AND INDUSTRIAL RESEARCH. Invention is credited to Jai Shankar Arya, Soumya Awasthi, Mahendra Pandurang Darokar, Subhas Chandra Gupta, Suman Preet Singh Khanuja, Sushil Kumar, Laiq-Ur Rahman, Ajit Kumar Shasany, Monika Singh.
Application Number | 20060057234 11/269594 |
Document ID | / |
Family ID | 35482513 |
Filed Date | 2006-03-16 |
United States Patent
Application |
20060057234 |
Kind Code |
A1 |
Khanuja; Suman Preet Singh ;
et al. |
March 16, 2006 |
Composition comprising pharmaceutical/nutraceutical agent and a
bio-enhancer obtained from Glycyrrhiza glabra
Abstract
The invention relates to a new use of a non-alkaloid compound
that is plant derived glycoside `glycyrrhizin` as a highly potent
bio-enhancer of activity and availability of antibiotics and other
drugs including anti-infective and anticancer agents. The molecule
of invention facilitates the absorption/uptake of antibiotics and
other molecules across the cell membrane in plant and animal cells
as well as Gram-positive and Gram-negative bacteria and therefore
can be used as a drug facilitator or bioenhancer molecule to
increase the affectivity of drugs and nutraceutical agents. The
compound having no antimicrobial or cytotoxic activity of its own,
is a safe candidate to reduce the drug dosage towards circumventing
the problem of drug resistance and the other side effects in
anti-infective and anti-cancer therapies.
Inventors: |
Khanuja; Suman Preet Singh;
(Lucknow, IN) ; Kumar; Sushil; (Lucknow, IN)
; Arya; Jai Shankar; (Lucknow, IN) ; Shasany; Ajit
Kumar; (Lucknow, IN) ; Singh; Monika;
(Lucknow, IN) ; Awasthi; Soumya; (Lucknow, IN)
; Gupta; Subhas Chandra; (Lucknow, IN) ; Darokar;
Mahendra Pandurang; (Lucknow, IN) ; Rahman;
Laiq-Ur; (Lucknow, IN) |
Correspondence
Address: |
FOLEY AND LARDNER LLP;SUITE 500
3000 K STREET NW
WASHINGTON
DC
20007
US
|
Assignee: |
COUNCIL OF SCIENTIFIC AND
INDUSTRIAL RESEARCH
|
Family ID: |
35482513 |
Appl. No.: |
11/269594 |
Filed: |
November 9, 2005 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
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09655710 |
Sep 5, 2000 |
6979471 |
|
|
11269594 |
Nov 9, 2005 |
|
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Current U.S.
Class: |
424/757 ;
514/154; 514/192; 514/200; 514/253.08; 514/291; 514/3.1;
514/312 |
Current CPC
Class: |
A61K 36/484 20130101;
A61K 45/06 20130101; A61K 36/484 20130101; A61K 2300/00
20130101 |
Class at
Publication: |
424/757 ;
514/253.08; 514/312; 514/008; 514/192; 514/200; 514/154;
514/291 |
International
Class: |
A61K 36/484 20060101
A61K036/484; A61K 31/496 20060101 A61K031/496; A61K 31/4709
20060101 A61K031/4709 |
Claims
1. A composition comprising an effective amount of an extract or
compound obtained from the plant Glycyrrhiza glabra and useful as a
bio-enhancer and bioavailability facilitator together with a
therapeutically effective amount of one or more nutraceuticals,
antibiotics, anti-infective agents and anti-cancer agents.
2-4. (canceled)
5. A composition as claimed in claim 1 wherein the anti-infective
agents are selected from anti-bacterial, anti-fungal and
anti-tuberculosis agents.
6. (canceled)
7. A composition as claimed in claim 1 wherein the concentration of
glycyrrhizin ranges from 0.10 to 10% of the weight of the
nutraceutical compounds.
8. A composition as claimed in claim 1 wherein the concentration of
glycyrrhizin ranges from 0.25 to 20% of the weight of the
anti-infective agent.
9. A composition as claimed in claim 5 wherein the anti-bacterial
agents are selected from the group comprising P-lactams,
macrolides, quinolones, fluoro-quinolones, aminoglycosides,
glycopeptides, rifamycins, folate inhibitors, tetracyclines,
anti-tuberculosis agents and microbicidal compounds.
10. A composition as claimed in claim 9 wherein the quinolones and
fluoro-quinolones are selected from the group consisting of
nalidixic acid, norfloxacin, ciprofloxacin, sparfloxacin and
similar other compounds.
11. A composition as claimed in claim 9 wherein the .beta.-Lactams
are selected from the group comprising penicillin, flucloxacillin,
cloxacukkub, methicillin, cephalosporins and carbapenenms.
12. A composition as claimed in claim 9 wherein the macrolides are
selected from erythromycin, clarithromycin, azithromycin and other
similar compounds.
13. A composition as claimed in claim 9 wherein the aminoglycosides
are selected from the group consisting of streptomycin, gentamicin,
amikacin and similar compounds.
14. A composition as claimed in claim 9 wherein the glycopeptides
are selected from vancomycin, teicoplanin and other similar
compounds.
15. A composition as claimed in claim 9 wherein the rifamycins are
rifunpicin and similar compounds.
16. A composition as claimed in claim 9 wherein the folate
inhibitors are selected from trimethoprim, sulphonamides and other
similar compounds.
17. A composition as claimed in claim 9 wherein the microbiocidal
compounds are selected from streptogramins and oxazolidinones.
18. A composition as claimed in claim 5 wherein the
anti-tuberculosis agents are selected from the group comprising
isoniazid, pyrazinamide, ethambutol and other similar
compounds.
19. A composition as claimed in claim 5 wherein the anti-fungal
agents are selected from the group consisting of polyene, triazole,
imidazole, clotrimazole and other fungicidal compounds.
20. A composition as claimed in claim 19 wherein the polyene is
selected from amphoterecin B, nystatin and other similar
compounds.
21. A composition as claimed in claim 19 wherein the triazole is
selected from fluconazole, itraconazole and other similar
compounds.
22. A composition as claimed in claim 19 wherein the imidazole is
selected from ketoconazole and other similar compounds.
23. A composition as claimed in claim 19 wherein the fungicidal
compound is selected from griseofulvin and terbinafine.
24. A composition as claimed in claim 1 wherein the anti-cancer
agent is selected from the group consisting of paclitaxel,
docetaxel, vinblastine, vincristine, and vinorelbine.
25. A composition as claimed in claim 1 wherein the concentration
of giycyrrhizin ranges from 10 to 10,000 fold folds of the weight
of the anti-cancer compound.
26. A composition as claimed in claim 1 wherein the nutraceutical
compounds are selected from the group consisting of vitamins, amino
acids, hormones and other nutritional supplements.
27. (canceled)
28. A method for enhancing the bio-availability of a herbal
extract, ant-infective agent, anti-cancer agent or nutraceutical
compound across biological membranes in a patient in need thereof
comprising administering to the patient composition comprising an
effective amount of a bioenhancer obtained from the plant
Glycyrrhiza glabra, and any one of an anti-infective agent,
anti-cancer agent or nutraceutical compound.
29. A method as claimed in claim 28 wherein the bio-enhancer is
selected from the extract of Glycyrrhiza glabra, glycyrrhizin,
glycyrrhizic acid obtained from Glycyrrhiza glabra or combinations
thereof.
30. A method as claimed in claim 28 wherein the bioenhancer is
obtained from the roots, stem, stolon or leaves of the plant
Glycyrrhiza glabra.
31. A method as claimed in claim 28 wherein the anti-infective
agent is an anti-bacterial agent anti-fungal agent or anti-cancer
agent.
32. A method as claimed in claim 31 wherein the anti-bacterial
agent is selected from the group consisting of .beta.-lactams,
macrolides, quinolones, fluoro-quinolones, aminoglycosides,
glycopeptides, rifamycins, folate inhibitors, tetracyclines,
anti-tuberculosis agents and microbicidal compounds.
33. A method as claimed in claim 31 wherein concentration of
glycyrrhizin ranges from 0.05 to 50% the weight of the
anti-bacterial agent.
34. A method as claimed in claim 32 wherein the quinolones and
fluoroquinclones are selected from the group consisting of
nalidixic acid, norfloxacin, ciprofloxacin, and sparfloxacin.
35. A method as claimed in claim 32 wherein the .beta.-Lactams are
selected from the group comprising penicillin, flucloxacillin,
cloxacukkub, methicillin, cephalosporins and carbapenenms.
36. A method as claimed in claim 32 wherein the macrolides are
selected from the group consisting of erythromycin, clarithromycin,
and azithromycin.
37. A method as claimed in claim 32 wherein the aminoglycosides are
selected from the group consisting of streptomycin, gentamicin, and
amikacin.
38. A method as claimed in claim 32 wherein the glycopeptides are
selected from vancomycin, and teicoplanin.
39. A method as claimed in claim 32 wherein the rifamycins are
rifampicin.
40. A method as claimed in claim 32 wherein the anti-tuberculosis
agents selected from the group consisting of comprising isoniazid,
pyrazinamide, and ethambutol.
41. A method as claimed in claim 31 wherein concentration of
glycyrrhizin ranges from 0.25 to 20% the weight of the anti-fimgal
agent.
42. A method as claimed in claim 31 wherein the anti-fungal agent
is selected from the group consisting of polyene, triazole,
imidazole, and clotrimazole.
43. A method as claimed in claim 28 wherein the anti-cancer agent
is selected from the group consisting of paclitaxel, docetaxel,
vinblastine vincristine, and vinorelbine.
44. A method as claimed in claim 31 wherein the concentration of
glycyrrhizin ranges from 10 to 10,000 fold of the weight of the
anti-cancer agent.
45. A method as claimed in claim 28 wherein the nutraceutical
compound is selected from the group consisting of vitamins, amino
acids, hormones and other nutritional supplements.
46. A method as claimed in claim 28 wherein administration of the
composition circumvents the side effects of chemotherapy by
substantially reducing the dosage of the anti-infective or
anti-cancer agent.
47. A method as claimed in claim 28 wherein the bioenhancer
improves the uptake of anti-infectives, anticancer agents and other
molecules when glycyrrhizin is provided prior to the treatment with
these molecules.
48. (canceled)
49. A method as claimed in claim 46, wherein the side effect is
bone marrow suppression, liver toxicity, kidney toxicity, pulmonary
scarring, high fever, skin reaction, nausea, vomiting, hair loss,
skin rash, mouth sores, diarrhea, loss of appetite, confusion,
lethargy, ambulation difficulty, ataxia, an anaphylactic reaction,
fast heart rate, wheezing, lowered blood pressure, facial edema,
cerebellar dysfunction, slurring of speech, walking problems, eye
motion problem, loss of periods, menses, ovarian failure, soft
tissue ulcers, cardiotoxicity, a hypersensitivity reaction,
abnormal blood pressure, sweating, bleeding, or shortness of
breath.
Description
FIELD OF THE INVENTION
[0001] The invention provides novel compositions containing
extracts and compounds obtained from the plant Glycyrrhiza glabra,
useful as bio-enhancers and bioavailability facilitators for a
variety of molecules including anti-infective, anti-cancer agents
and nutritional compositions. This invention also relates to a
method for enhancing the biological availability, proficiency and
effectivity of drugs, pharmaceuticals, nutraceuticals and other
related compounds including amino acids, vitamins and other
nutritional elements and ions. The present invention has direct
implication in drastically reducing the dosage of antibiotics,
drugs and anti-cancer agents while increasing the efficiency of
absorption of nutritional elements.
[0002] 1. Background of the Invention
[0003] Since the invention of penicillin in 1928 by Alexander
Flemming and their wider availability in the 1940s, further
variants of existing and new antibiotics have been on the trial.
Compared to the situation in 1954 when two million pounds of
antibiotics were produced in the US, the present figure exceeds 50
million pounds. Antibiotics work either by killing bacteria
(bacteriocidal) or by inhibiting growth (bacteriostatic) and their
bioactivities have transformed human ability to treat many
infectious diseases that previously were considered certain
killers. It is estimated that globally humans consume above 250
million doses of antibiotics annually and 20%-50% of that use is
unnecessary depending on the class of antibiotic. Further,
widespread use of antibiotics promotes spread of antibiotic
resistance many a times leading to multiple drug resistance. The
total amount of antibiotic in the given dosage used for treatment
of an infection generally is much higher than what is actually
required to control a given population of parasite in the infected
individual. This is so because all antibiotic given to the patient
in a therapy does not reach the target site. This may be due to (i)
lower absorption in the gut membrane when taken orally (ii)
restrictive uptake by the target microbe or (iii) operation of
efflux pump leading to indiscriminate extrusion of the antibiotics
or therapeutic molecules. Thus, large portions of the drugs we
apply are wasted and only a miniscule percentage is being targeted
to the infective microbes. But even worse part is that the
unutilized drug/antibiotic amount remains as a load in the body and
environment. This then acts as a selection pressure, facilitating
emergence of drug resistance in parasites leading to their
predominance in the niche and ultimately resulting into failure of
antibiotics against resistant infections. Additionally, such a
situation leads to side effects, illness and reduction in life
expectancy being more acute in the older population. One of the
ways that has been feasible to reduce drug dosage is the occurrence
of synergism between different therapeutic agents. However, even in
such a situation if both the molecules have the antibiotic
property, the problem of continued selection pressure on microbes
is still likely to continue. Therefore, the need is for molecules,
which by themselves are not microbicidal but when present with a
drug or active molecule, enhance its activity and availability
(bioenhancers). These molecules by their presence will not exert
any selection pressure for the mutants to emerge resistant against
them and on the other hand could reduce the dosage of antibiotics
or drugs so that their ill effects are minimized. This way
resistance development process will be substantially delayed
ultimately leading to enhanced life-span of the novel and existing
antibiotics. Such drug/molecule facilitators should have novel
properties like non-toxic to humans, animals or plants, should be
effective at a very low concentration in a combination, should be
easy to formulate and most importantly enhance uptake/absorption
and activity of the drug molecules. This can lead to development of
judicious and strategic concentrations of antibiotics with specific
bioenhancers to improve availability of the drug right up to the
target for effectively controlling the infectious organisms.
[0004] The present invention was the result of planned experiments
to provide a novel method for improving activity and
bioavailability of antibiotics, drugs and other molecules using a
plant glycoside "Glycyrrhizin" in different formulations.
[0005] The bioavailability of nutrients and enhancement of
antibiotics/drugs effectivity is relevant to human, plant as well
as animal health and thus the compositions and methods of the
invention are also intended to be used in agriculture and
veterinary practice.
[0006] 2. Description of Related Art
[0007] Use of ayurvedic preparation "trikatu" dates back to the
period between the seventh century B.C. and the sixth century A.D.,
which is a Sanskrit word meaning three acrids. It refers to a
combination of three elements namely, black pepper (Piper nigrum
Linn.), long pepper (Piper longum Linn.) and ginger (Zingiber
officinale Rosc.). It is believed that the use of "trikatu", and
its constituents individually as well as collectively, enhances the
bioavailability of a number of drugs. In specific studies carried
out on animals as well as human volunteers, it was noted that the
active component responsible for the increase in bioavailability of
various drugs was piperine (U.S. Pat. Nos. 5,616,593 and
5,972,382). Till today, thus, the known documented bio-availability
enhancer is piperine and a series of inventions related to this
compound have been described by several prior art investigators.
Though the compound piperine has been reported to be enhancing the
bioavailability of drugs, nutrients and vitamins, still a proper
formulation for the combination is yet to come to the market.
Beside this, the compound piperine being an alkaloid would find
restricted use for the reasons of its typical action.
[0008] The Applicants sought to provide a non-alkaloid molecule
capable of bio-enhancing the availability of anti-infective,
nutritional and anti-cancer compounds like `taxol` produced by the
yew tree in microscopic amounts.
[0009] The Applicants screened a large number of the available
extracts and known compounds in the laboratory at the Central
Institute of Medicinal & Aromatic Plants, Lucknow, U.P., India
(CIMAP), particularly those, which by themselves did not exhibit
any antibacterial property. After extensive experimentation, it was
found that out of 70 pure compounds tested, only one plant compound
`glycyrrhizin`, a glycoside enhanced the killing activities of
different antibiotics on three diverse kinds of bacteria including
Escherichia coli, Bacillus subtilis and Mycobacterium smegmatis.
The compound is generally isolated from the roots and stolon of the
plant Glycyrrhiza glabra and is a known sweetener and
hepatoprotective drug.
[0010] Licorice (the extract from the root of the plant) is used to
flavor foods, tobacco, drinks, medicines and sweets. It has
expectorant action to treat bronchitis and can reduce inflammation,
allergies, asthma, gastritis, peptic ulcers, rheumatism, and sore
throat. It helps the liver to detoxify drugs and is used for
treatment of liver disease. Licorice strengthens the immune system,
stimulates the adrenal gland, and is diuretic and laxative.
Licorice from roots of Glycyrrhiza glabra, contains glycyrrhizin,
which is 50 times sweeter than sugar. Primary uses for licorice
include treatment for peptic ulcers and stomach ailments,
respiratory and intestinal passages. Licorice makes a wonderful
expectorant expelling mucus from the chest area.
[0011] The compound when studied by the Applicants also showed
enhancement in the activity of known azole class antifungal agents
like clotrimazole against infectious fungal strain of Candida
albicans. More important was the obvious enhancement in the cell
division inhibitory activity against the breast cancer cell line
MCF-7.
[0012] Accordingly, the Applicants have sought to increase the
absorption of anti-infective, nutritional and anti-cancer compounds
by the body and increase their utilization by the body by providing
novel compositions containing extracts from the plant glycyrrhiza
glabra.
OBJECTS OF THE INVENTION
[0013] The main object of the invention is to provide novel
compositions comprising extracts and compounds from the plant
Glycyrrhiza glabra useful as a bioenhancer and capable of improving
or enhancing the bio-availability of drugs such as antibiotics,
anti-infective agents and of nutritional compounds.
[0014] Another object of the invention is to provide compositions
comprising extracts and compounds from the plant Glycyrrhiza glabra
useful as bio-enhancers and bioavailability facilitators, thus
enabling effective transport of drugs and molecules across
biological membranes to reach target sites.
[0015] Still another object is to provide methods for enhancing the
bio-availability of drugs molecules and nutritional to the target
site in the body.
SUMMARY OF THE INVENTION
[0016] Accordingly, the invention provides novel compositions
containing extracts and compounds from the plant Glycyrrhiza glabra
useful as bio-enhancers and bio-availability facilitators for drugs
and molecules such as antibiotics, anti-infective agents,
anti-cancer agents and for nutritional compounds. The invention
also provides methods for enhancing the availability of drugs,
molecules and nutritional compounds to effectively reach the target
site in the body.
DETAILED DESCRIPTION
[0017] Accordingly, the invention provides a composition comprising
an effective amount of an extract or compound obtained from the
plant Glycyrrhiza glabra and useful as a bio-enhancer and
bioavailability facilitator together with a therapeutically
effective amount of one or more nutraceuticals, antibiotics,
anti-infective agents and anti-cancer agents.
[0018] In an embodiment, the composition contains an extract from
the leaves, stem, roots or stolon of Glycyrrhiza glabra,
glycyrrhizic acid or glycyrrhizin, or a combination thereof
[0019] In still another embodiment, glycyrrhizin and glycyrrhizic
acid are obtained from the plant Glycyrrhiza glabra.
[0020] In yet another embodiment, the anti-infective agents are
selected from anti-bacterial, anti-fungal and anti-tuberculosis
agents.
[0021] In yet another embodiment, the extract from the roots of
Glycyrrhiza glabra is licorice. In an embodiment, the compound as
bioavailability enhancer facilitates the transport of the
compound(s) across the biological membranes to reach the target
site for better effectivity/activity.
[0022] In an embodiment, the concentration of glycyrrhizin ranges
from 0.05 to 50% of the weight of the anti-bacterial compounds.
[0023] In still another embodiment, the concentration of
glycyrrhizin ranges from 0.10 to 10% of the weight of the
nutraceutical compounds.
[0024] In yet another embodiment, the concentration of glycyrrhizin
ranges from 0.25 to 20% of the weight of the anti-fungal
agents.
[0025] In another embodiment, the anti-bacterial agent is selected
from the group consisting of .beta.-lactams, macrolides,
quinolones, fluoro-quinolones, aminoglycosides, glycopeptides,
rifamycins, folate inhibitors, tetracyclines, anti-tuberculosis
agents and other similar microbicidal compounds.
[0026] In still another embodiment, the anti-fungal agent used is
selected from the group comprising of polyene, triazole, imidazole,
clotrimazole and other fungicidal compounds. In still another
embodiment of the present invention bioenhancer improves the uptake
of anti-infectives, anticancer agents and other molecules when
glycyrrhizin is provided prior to the treatment with these
molecules.
[0027] In yet another embodiment of the present invention the
anticancer agents include paclitxel (taxol) or other cell division
inhibitors controlling the growth and multiplication of
tumor/cancer cells.
[0028] In yet another embodiment of the present invention living
system used is selected from bacteria, fungi or any living
cells.
[0029] In yet another embodiment of the present invention the
bioenhancer element is used in pure form as glycyrrhizin or as
precursor/derivatives like glycyrrhizic acid.
[0030] In yet another embodiment of the present invention the
bioenhancer element is administered provided as licorice in form of
aquous/alcoholic extracts, decoction or direct root
flakes/chips/sticks through wetting by any means.
[0031] In yet another embodiment of the present invention a
pharmaceutical composition to increase the bioactivity and
bioavailability comprising of the plant compound glycyrrhizin and a
bio-active chemical compound selected from the group consisting of
anti-bacterial, anti-fungal, anti-cancerous and other similar
compounds including nutraceuticals wherein, the glycyrrhizin
concentration ranges from 0.10 to 10% of the weight of the
nutraceuticals compound used.
[0032] In yet another embodiment, the anti-bacterial agent is
selected from the group consisting of .beta.-lactams, macrolides,
quinolones, fluoro-quinolones, aminoglycosides, glycopeptides,
rifamycins, folate inhibitors, tetracyclines, anti-tuberculosis
agents and other similar microbicidal compounds wherein, the
glycyrrhizin concentration ranges from 0.05 to 50% of the weight of
the anti-bacterial compound used.
[0033] In yet another embodiment, the macrolides comprise
erythromycin, clarithromycin, azithromycin and other similar
compounds.
[0034] In yet another embodiment, the quinolones and
fluoroquinolones used are selected from the group consisting of
nalidixic acid, norfloxacin, ciprofloxacin, sparfloxacin and
similar other compounds.
[0035] In yet another embodiment of the present invention the
aminoglycosides used are selected from the group consisting of
streptomycin, gentamicin, amikacin and other similar compounds.
[0036] In yet another embodiment of the present invention the
glycopeptides used are vancomycin, teicoplanin and other similar
compounds.
[0037] In yet another embodiment of the present invention the
rifamycins used are rifampicin and other similar compounds.
[0038] In yet another embodiment of the present invention the
anti-tuberculosis agents used are isoniazid, pyrazinamide,
ethambutol and other similar compounds.
[0039] In yet another embodiment of the present invention the
folate inhibitors are trimethoprim; sulphonamides and other similar
compounds.
[0040] In yet another embodiment, the .beta.-Lactams comprise
penicillin, flucloxacillin, cloxacukkub, methicillin,
cephalosporins, carbapenenms and similar compounds.
[0041] In yet another embodiment, other similar microbicidal
compounds are streptogramins and oxazolidinones.
[0042] In another embodiment the anti-fungal agent is selected from
the group consisting of polyene, triazole, imidazole, clotrimazole
and other fungicidal compounds wherein, the glycyrrhizin
concentration ranges from 0.25 to 20% of the weight of the
anti-fungal compound used.
[0043] In yet another embodiment the polyenes used are amphoterecin
B, nystatin and other similar compounds.
[0044] In yet another embodiment the triazoles used are
fluconazole, itraconazole and other similar compounds.
[0045] In yet another embodiment the imidazoles used are
ketoconazole and other similar compounds.
[0046] In yet another embodiment other fungicidal compounds are
griseofulvin and terbinafine.
[0047] In yet another embodiment, the anti-cancer agent is selected
from the group consisting of Paclitaxel (Taxol), Docetaxel
(Taxotere), Vinblastine (Velban), Vincristine (Oncovin),
Vinorelbine (Navelbine) and other similar compounds used in cancer
therapy.
[0048] In another embodiment, the levels of glycyrrhizin ranges
from 10 to 10,000 folds of the weight of the anti-cancer compound
used.
[0049] In another embodiment, the nutraceuticals used to increase
the bio-activity and bio-availability are selected from the group
consisting of vitamins, amino acids, hormones and other nutritional
supplements.
[0050] In yet another embodiment, the pharmaceutical compositions
comprising of the plant compound glycyrrhizin in form of
liquorice/dried root with the bioactive chemical compound(s) to
increase their bioactivity and bioavailability wherein, the
glycyrrhizin in form of liquorice preparation or dried root is
given as one gram equivalent to 40 mg of glycyrrhizin.
[0051] The invention also provides a method for enhancing the
bio-availability of herbal extracts, anti-infective agents,
anti-cancer and nutraceutical compounds/compositions across
biological membranes in living systems; said method comprising the
steps of administering to a subject in need thereof a composition
comprising an effective amount of the bioenhancer obtained from the
plant Glycyrrhiza glabra, and any one of anti-infective agents,
anti-cancer agents and nutraceutical compounds.
[0052] In an embodiment, the bio-enhancer is selected from the
extract of Glycyrrhiza glabra, glycyrrhizin, glycyrrhizic acid
obtained from Glycyrrhiza glabra or combinations thereof
[0053] In yet another embodiment, administration of the composition
circumvents the side effects of chemotherapy by substantially
reducing the dosages of therapeutic agents comprising of
anti-bacterial, anti-fungal and anti-cancerous agents wherein the
side effects of those agents can be bone marrow suppression, liver
and kidney toxicity, pulmonary scarring, high fevers, skin
reactions, Nausea and vomiting, hair loss, skin rash, mouth sores,
diarrhoea, loss of appetite and confusion, lethargy and ambulation
problems (ataxia), anaphylactic reactions (consisting of fast heart
rate, wheezing, lowered blood pressure and facial edema, cerebellar
dysfunction (consists of slurring of speech, walking problems, as
well as eye motion problems), loss of periods (menses), ovarian
failure can occur, painful soft tissue ulcers, cardiotoxicity,
hypersensitivity reactions, abnormal blood pressure, sweating,
bleeding, shortness of breath and many related problems.
[0054] In still another embodiment, the bioenhancer improves the
uptake of anti-infectives, anticancer agents and other molecules
when glycyrrhizin is provided prior to the treatment with these
molecules.
[0055] The invention also provides use of the extract or the
compound obtained from the plant Glycyrrhiza glabra, as a
bioenhancer and bioavailability and facilitator of nutritional
compounds and drugs and molecules selected from anti-infective and
anti-cancer agents.
[0056] The invention is described in detail herein below with
reference to the following drawings wherein:
[0057] FIG. 1: represents the structural formula of Glycyrrhizic
acid; and
[0058] FIG. 2: represents the structural formula of
Glycyrrhizin
[0059] Some of the embodiments of the invention are illustrated
herein below by the following bioassays and examples which are
provided merely to illustrate the invention and should not be
construed as limitations on the inventive concept embodied
herein.
[0060] The methodology followed for screening of this compound
(Glycyrrhizic acid and Glycyrrhizin) included specifically designed
bioassays as follows:
1. Assay for Bio-Enhancement of Anti-Infective Agents
[0061] a) The minimum inhibitory concentration (MIC) of any
antibiotic or drug molecule is determined against Escherichia coli
(ATCC 10536), Bacillus subtilis (ATCC 6015 ) and Mycobacterium
smegmatis (ATCC 14468) and Candida albicans (ATCC 10231) in broth
and disc diffusion assays. [0062] b) The antibiotics or antifungal
agents at concentrations 1/10, 1/8, 1/6, 1/4, 1/3, 1/2 and equal to
MIC are added alone and in combination with the test compound at
varying concentrations in broth assays to evaluate the comparative
level of inhibition and similarly at varying concentrations in disc
diffusion assay. [0063] c) The combinations showing enhancement in
terms of higher activity than antibiotic alone for the inhibition
of bacterial growth (large inhibition zone in disc diffusion and
higher killing at lower concentration in broth assay) are picked up
for future testing.
[0064] d) In broth assays the activity is quantified by counting
number of viable cells in a given treatment and converted in fold
enhancement by the combination treatment compared to
antibiotic/drug alone in the killing percentage of cells.
[0065] e) The pretreatment assay is followed to determine whether
the bioenhancer which may be either an extract obtained from the
plant Glycyrrhiza glabra, glycyrrhizin or glycyrrhizic acid, is
required along with antibiotic to enhance its activity or even its
withdrawal after treatment or prior to antibiotic treatment would
benefit. For this, the cells are treated with the
bioenhancer/compound for 4 to 8 hours and then washed free of it by
centrifugation and washing in sterile water. This is followed by
treatment with antibiotic/drug as in steps (b) to (d) above.
[0066] f) The results of the bio-assays carried out are represented
in the tables provided hereinbelow wherein tables 1-4 represent the
activity of glycyrrhizin. Tables 7-9 depict the activity of
glycyrrhizic acid and tables 10-11 depict the activity of
licorice.
2. Assay for Bio-Enhancement of Anti-Cancer Agents
[0067] a) MCF-7 (Breast cancer cell line is inoculated at a density
of about 0.1.times.10.sup.6 cells in MEM medium in the wells of 24
well plate. [0068] b) The used up medium in these cultures is
replaced with fresh medium after 18 hours in each well. [0069] c)
The test compound (s) is added at desired concentrations in
different wells just after the medium replacement.
[0070] d) Observations are recorded on the cell count after 36
hours for which the following steps are required. [0071] i. The
medium is removed from the wells. [0072] ii. The wells are rinsed
with 1 ml PBS (Phosphate buffer saline). [0073] iii. To each well
500 .mu.l of freshly prepared trypsin solution (0.1% in PBS) is
added. [0074] iv. Trypsin solution is removed after 30 seconds and
the plate is gently tapped till the cells are released from the
plate surface. [0075] v. Fresh 1 ml of MEM growth medium is added
and agitated with a pipette to obtain a cell suspension. [0076] vi.
10 .mu.l of cell suspension is taken on the haemocytometer and a
cover glass is placed over the counting chamber. [0077] vii. The
number of viable cells is counted in 5 big squares and the reading
is taken from 5 microscopic fields to determine the average.
[0078] viii. The cell count (titre per ml) in the original sample
is then calculated as average count.times.10.sup.3. TABLE-US-00001
Composition of Minimum Essential Medium (MEM): 100 ml MEM powder
(Sigma - Aldrich, USA) = 0.96 g HEPES Buffer (Sigma - Aldrich, USA)
= 0.26 g Sodium Bicarbonate = 0.22 g Penicillin G = 10 mg
Streptomycin = 20 mg Gentamycin = 5 mg Foetal Calf Serum = 15 ml
Foetal Calf Serum = 15 ml Distilled water = 85 ml
3. Bioavailability Tests Through Biological Membrane [0079] a) A
specially designed U-tubes of glass consisting of two component
(opposite-L type) were used in which one open end of a L-shaped was
tapered to fit within the untapered end of the other L-tube. [0080]
b) The membrane of goat gut (initial part) was stretched and fixed
to act as the barrier between the two ends such that by joining the
two L-tubes, a U-tube was made. [0081] c) Sterile distilled water
was then filled in both the sides to equal height/level. [0082] d)
The antibiotic/compound was added to the donor tube (tapered) and
through spectro-photometer the transfer of molecule was observed
using UV and visible absorption maxima of the respective molecules
by taking the OD at defined wavelengths.
EXAMPLES
[0083] In the next step of elucidation of the enhancer action the
applicants experimented with the killing activities of different
antibiotics against the bacteria singly and in combination with the
test compound glycyrrhizin following the method described above.
These experiments are to be considered with assay for
bioenhancement of the compounds glycyrrhizin/glycyrrhizic acid and
licorice, as described earlier. When the bacteria were grown in
presence of the compound as such no significant killing was
observed. In all the experiments the glycyrrhizin concentration was
kept at 1 .mu.g/ml, unless it is specifically mentioned.
Example 1
Glycyrrhizin Mediated Enhancement in the Killing Action of
Antibiotics Against Gram-Negative Bacterium Escherichia coli
[0084] TABLE-US-00002 TABLE 1 Survival Survival fraction *Folds
fraction of viable cells enhancement of viable cells upon treatment
in Concen- upon treatment with antibiotic- antibiotic tration with
antibiotic glycyrrhizin activity Antibiotics .mu.g/ml alone (a)
combination (b) (a/b) Rifampicin 10 0.42 0.11 3.8 Rifampicin 20
0.28 0.02 14.0 Nalidixic 8 0.19 0.02 9.5 acid Tetracycline 2 0.42
0.22 1.9 Tetracycline 4 0.27 0.03 9.0 Ampicillin 6 0.38 0.03 12.6
Ampicillin 8 0.013 0.006 2.1
Example 2
Glycyrrhizin mediated enhancement in the killing action of
antibiotics against Gram-positive bacterium Bacillus subtilis
[0085] TABLE-US-00003 TABLE 2 Survival Survival fraction fraction
of of viable cells *Folds viable cells upon treatment enhancement
Concen- upon treatment with antibiotic- in antibiotic tration with
antibiotic glycyrrhizin activity Antibiotics .mu.g/ml alone (a)
combination (b) (a/b) Nalidixic 6 0.50 0.11 4.50 acid Nalidixic 8
0.27 0.04 6.75 acid Tetracycline 1 0.10 0.02 5.00
Example 3
Glycyrrhizin Mediated Enhancement in the Killing Action of
Antibiotics Against Bacterium Mycobacterium smegmatis
[0086] TABLE-US-00004 TABLE 3 Survival Survival fraction fraction
of of viable cells *Folds viable cells upon treatment enhancement
Concen- upon treatment with antibiotic- in antibiotic tration with
antibiotic glycyrrhizin activity Antibiotics .mu.g/ml alone (a)
combination (b) (a/b) Rifampicin 0.5 0.56 .times. 10.sup.-4 0.086
.times. 10.sup.-4 6.5 Nalidixic 4 0.70 0.12 7.7 acid Tetracycline
0.5 0.3 0.2 1.5
[0087] From the above experiments it was deduced that the potency
of the antibiotic is increased when applied along with the compound
glycyrrhizin. It was interesting to note that the same results were
observed when the bacterial cells were pretreated with glycyrrhizin
for 4-8 hours and then the cells were washed free of the
glycyrrhizin by centrifugation and resuspending the cells in fresh
medium followed by treatment with respective antibiotics. This
implicated some changes induced by glycyrrhizin on the bacterial
surface/membrane making it more receptive to the antibiotics and
hence enhancing bioavailability and ultimately
activity/bactericidal effect of the given antibiotic.
Example 4
Glycyrrhizin Mediated Enhancement Observed in Disc Diffusion Assays
for the Killing Action of Antifungal Agent Against Candida
albicans
[0088] TABLE-US-00005 TABLE 4 Clotrimazole Glycyrrhizin Zone of
inhibition (.mu.g/disc) (.mu.g/disc) (mm) 0 0 0 0 1 0 4 0 7 4 1
11
Example 5
Glycyrrhizin Mediated Enhancement in the Activity of Anticancerous
Agent "Taxol"
[0089] TABLE-US-00006 TABLE 5 Final titre of Final titre of viable
cells Taxol Initial titre viable cells upon treatment with
Concentration of viable upon treatment taxol-glycyrrhizin
(.mu.g/ml) cells with taxol alone combination 0.01 0.1 .times.
10.sup.-6 0.05 .times. 10.sup.6 0.025 .times. 10.sup.6 0.02 0.1
.times. 10.sup.-6 0.04 .times. 10.sup.6 0.024 .times. 10.sup.6 0.05
0.1 .times. 10.sup.-6 0.03 .times. 10.sup.6 0.021 .times. 10.sup.6
Without taxol Final titre Control 0.1 .times. 10.sup.-6 0.16
.times. 10.sup.-6 Control 0.1 .times. 10.sup.-6 0.12 .times.
10.sup.-6 (glycyrrhizin)
Example 6
Glycyrrhizin Mediated Enhancement in the bioavailability Across the
Biological Membrane
[0090] TABLE-US-00007 TABLE 6 OD measured as Absorbance Wave
(specific to the compound maxima) Compound(s) in length across the
membrane in receiving tube after the donor tube (nm) 1 hr 2 hr 3 hr
4 hr 5 hr 6 hr Rifampicin A.sub.340 0.0039 0.0275 0.0371 0.0440
0.0906 0.1165 A.sub.475 0.0015 0.0148 0.0210 0.0297 0.0334 0.0394
Rifampicin + Glycyrrhizin A.sub.340 0.0070 0.0638 0.1465 0.2090
0.3424 0.3930 A.sub.475 0.0100 0.0395 0.0575 0.1362 0.2365 0.2853
Vitamin B-12 A.sub.360 0.0250 0.0556 0.0714 0.1283 0.1370 0.1595
A.sub.530 Vitamin B-12 + Glycyrrhizin A.sub.360 0.0582 0.0627
0.1058 0.2051 0.2616 0.2984 A.sub.530
[0091] Here, the applicants observed glycyrrhizin-mediated
enhancement in the cell division inhibitory action of anti-cancer
agent taxol in the animal cell culture experiments using cancerous
cell line MCF-7. As evident in Table 5 above, the anticancerous
activity of `Taxol` in terms of inhibiting the growth and
multiplication of MCF-7 cancer cells was markedly enhanced by at
least 5 folds. In these experiments, the cancerous cells growth
inhibition by Taxol (@ 0.01 .mu.g/ml) in presence of glycyrrhizin
(@ 1 .mu.g/ml) was higher than even the treatment with taxol (@
0.05 .mu.g/ml) alone.
Example 7
Glycyrrhizic Acid Mediated Enhancement in the Killing Action of
Antibiotics Against Escherichia coli in Broth Assay
[0092] TABLE-US-00008 TABLE 7 Survival fraction Survival of viable
cells *Folds fraction upon treatment en- of viable cells with
antibiotic hancement Concen- upon treatment glycyrrhizic acid in
tration with antibiotic combination antibiotic Antibiotics .mu.g/ml
alone (a) (b) activity (a/b) Rifampicin 20 0.57 0.281 2.03
[0093] TABLE-US-00009 TABLE 8 Glycyrrhizic acid mediated
enhancement in the killing action of antibiotics against
Escherichia coli in disc diffusion assay. Glycyrrhizic Net zone of
Rifampicin (.mu.g/disc) acid (.mu.g/disc) inhibition (mm) 0 0 0 0 4
0 500 0 2 500 4 3
[0094] TABLE-US-00010 TABLE 9 Glycyrrhizic acid mediated
enhancement in the killing action of antibiotics against
Mycobacterium smegmatis in disc diffusion assay. Rifampicin
Glycyrrhizic Net zone of (.mu.g/disc) acid (.mu.g/disc) inhibition
(mm) 0 0 0 0 4 0 0.4 0 3 0.4 4 4
[0095] In the above experiments, the Applicants used Glycyrrhizic
acid to test the potency of this acid in enhancing the activity of
antibiotics. The Applicants observed that the killing action the
antibiotics (such as Rifampicin) is enhanced when used with
Glycyrrhizic acid. Table 7 shows that when the cells were treated
with Rifampicin alone, the survival fraction was about 0.57.
However, the survival fraction of cells when treated with a
combination of rifampicin and glycyrrhizic acid was much less i.e.
about 0.281. This shows that the combination of glycyrrhizic acid
with rifampicin enhances the availability of rifampicin to the
cells. Similarly, the results of the disc diffusion assay also show
that the combination of glycyrrhizic acid with rifampicin is more
effective against mycobacterium smegmatis than glycyrrhizic acid or
rifampicin used singly.
Example 8
Licorice (25 (.mu.g/ml) Mediated Enhancement in the Killing Action
of Rifampicin Against Gram-Negative Bacterium Escherichia coli
[0096] TABLE-US-00011 TABLE 10 Survival Survival fraction *Folds
fraction of of viable cells en- viable cells upon treatment
hancement Concen upon treatment with antibiotic in antibiotic
tration with antibiotic glycyrrhizic acid activity Antibiotics
.mu.g/ml alone (a) combination (b) (a/b) Rifampicin 10 0.420 0.135
3.10
[0097] TABLE-US-00012 TABLE 11 Licorice (25 .mu.g/ml) mediated
enhancement in the killing action of antibiotics against
Gram-positive bacterium Bacillus subtilus. Survival Survival
fraction fraction of of viable cells viable cells upon treatment
*Folds upon treatment with antibiotic enhancement in Concentration
with antibiotic glycyrrhizic acid antibiotic activity Antibiotics
.mu.g/ml alone (a) combination (b) (a/b) Nalidixic acid 6 0.500
0.121 4.10
[0098] Tables 10 and 11 show the bio-enhancing effects observed in
the use of licorice, the extract from the roots of Glycyrrhiza
glabra. As can be seen from Tables 10, the survival rate of
bacteria treated with the antibiotic Rifampicin is more than when
treated with a combination of Rifampicin and licorice. Similarly,
Table 11 shows that the survival rate of the bacteria treated with
nalidixic acid is less than when treated with a combination of
nalidixic acid and licorice. The results in Tables 10 and 11
demonstrate, the use of licorice as an effective bio-enhancer and
bioavailability facilitator.
[0099] In the experiments related to transport across the mammalian
gut membrane, it was observed that the compound glycyrrhizin
enhances the transport of antibiotics e.g. Rifampicin,
Tetracycline, Nalidixic acid, Ampicillin and vitamins like B-1 and
B-12 across the gut as well as artificial membrane. The enhancement
in transport is approximately 2 to 6 folds.
[0100] Several medicinal plants are cited for use in Indian system
of medicine for various activities such as immunostimulant,
adaptogenic, anticancer, antimalarial, antimicrobial, angiogenetic,
hepatoprotective, hypolinidemic, hypoglycemic, memory enhancing
etc. The genus Glycyrrhiza (Mulethi) has attracted sizable
importance for compound(s) with hepatoprotective ability. In
Glycyrrhiza glabra (family, Leguminosae) 12 different varieties
(Biovars) are known. Among these only three namely typica,
glandilifera and violacea are commonly used for the pharmaceutical
and confectionary requirements. Glycyrrhiza glabra var. typica is
the source of Spanish Liquorice and is sweeter than other types.
Glycyrrhiza glabra var. glandilifera is the source of Russian
liquorice, which is moderately sweet and some what bitter, also
more or less acrid. Glycyrrhiza glabra var. violacea is known as
the Persian or Iranian liquorice of commerce and has thick roots.
The name Glycyrrhiza (popularly known as mulethi in India) is of
Greek origin meaning sweet root. Cultivation of this species is
mostly limited to US, Eastern Europe and several CIS of former USSR
and scattered in western and northern India. The roots of the plant
Glycyrrhiza glabra are highly useful as these contain the important
glycoside Glycyrrhizin for its hepatoprotective ability. The root
extracts also contain flavonoids, saponins, sterols, amino acids,
gums essential oils, sugars and starches. The plant takes at least
4-5 years to achieve substantial production of glycyrrhizin in the
roots. Generating sufficient planting material for commercial
cultivation is thus desirable for harvesting required quantity of
Glycyrrhizin on sustainable basis.
[0101] This molecule (Glycyrrhizin) is a known sweetener and
hepatoprotective drug. It helps the liver to detoxify drugs and is
given for liver disease. The molecule is known to be strengthening
the immune system, stimulates the adrenal gland, and is diuretic
and laxative. This is also used to treat peptic ulcers and stomach
ailments, respiratory and intestinal passages. Considering the wide
use of the molecule in traditional system of medicine and the level
of toxicity well tested, this can be used along with the
antibiotics and other molecule of importance to enhance the
efficacy on the target. This molecule (glycyrrhizin) in the
Applicant's study facilitates the transport/uptake of various
compounds including antibiotics, antifungal and anticancer agents,
vitamins and amino acids etc. across the membrane thereby
increasing the absorption or bioavailability various biological
systems including bacteria, animal and plant cells. This thus has
immense importance for absorption of the drugs, pharmaceuticals,
nutraceutical and other related compounds and ions by the cells and
hence huge potential of being used as drug/molecule
facilitators.
[0102] Great emphasis now is being laid towards quality assurance
of crude drugs from plants sources widely used in the Indian system
of medicine. The scientific study of traditional medicines,
derivation of drugs through bioprospection and systematic
conservation, domestication and cultivation of the concerned
medicinal plants has assumed great importance in the present day
context when more and more people prefer safe and effective
medicines at affordable price for curing their ailments. The
present invention enlarges the scope and use of the natural
glycoside glycyrrhizin in therapeutical and nutraceutical
application. These plants synthesizing multi-use molecules are
Glycyrrhiza sps and Abarus sps. The preferred plant is Glycyrrhiza
glabra which is propagated through underground rhizome. This
demands large amount of planting material which may be possible by
micro propagation in vitro for its commercial cultivation.
Agrotechnology and suitable cultivars for its cultivation in India
are therefore required through screening of germplasm and genetic
selection. At CIMAP a genetically superior variety of Glycyrrhiza
glabra var. typica named as `Misree` is available for cultivation
with better yield of glycyrrhizin (Kukreja. A K, Tyagi B R, Muni
Ram and Sushil Kumar (1997) Liquorice cultivar--Mishree. Journal of
Medicinal and Aromatic Plants Sciences. 19: 732-733).
[0103] Procedure for isolation of quality glycyrrhizin the main
compound from the root extract. Of the plant Glycyrrhiza glabra
var. typica was also standardized. For this purpose we choose the
plant Glycyrrhiza glabra var. typica `Misree` (the released variety
of CIMAP) and commercially available plant material. Fresh roots
were harvested from the plant and shade dried. Commercial dried
roots were purchased from the market. The protocol followed for the
isolation was as follows:
[0104] Protocol: [0105] 1. Plant material (250 gram) was dipped in
900 ml of Butanol: glacial acetic acid ethyl alcohol (1: 1:1) for
48 hours. [0106] 2. This was filtered through filter paper and
concentrated in vacuums rota-vapour. [0107] 3. The solid material
thus obtained was dissolved in acetic acid (180 ml) and treated
with diethyl ether 2 litre (10 times, each with 200 ml) to remove
impurity as precipitate which was filtered out. [0108] 4. Then the
liquor in diethyl ether was evaporated to 250 ml and fractionated
through liquid--liquid fractionation twice in water, followed by
discarding the water fraction. [0109] 5. This diethyl ether
fraction was further concentrated in vacuum rotavapour to obtain
the compound. [0110] 6. The compound was hydrolyzed with 80 ml of
NaOH (NaOH 5% in Methaol) at 80.degree.-90.degree. C for 1 hour.
[0111] 7. The hydrolyzed reaction mixture was acidified (100 ml of
glacial acetic acid acetic acid and 5 ml of concentrated HCl) to pH
3.6 to form a ppt which was filtered and washed with ethanol
[0112] After isolation, the compound was checked for the purity
compared to the standard glycyrrhizin obtained from Sigma chemical
Inc.
[0113] Preparation of the test solution: One gram of the isolated
compound was extracted in 20 ml chloroform for 15 min. After
filtration the filtrate was discarded. The marc was refluxed for 1
hour with 30 ml of 0.5 M H.sub.2SO.sub.4. The unfiltered mixture
was then cooled and extracted with chloroform (2.times.20ml) and
the combined CHCl.sub.3 extract was concentrated. The residue was
dissolved in 1.0 ml CHCl.sub.3: Methanol (1:1) mixture.
[0114] Reference solution: Five milligram of glycyrrhizin was
refluxed with 20 ml 0.5 M H.sub.2SO.sub.4. This was cooled and
extracted with chloroform (2.times.10 ml). The combined CHCl.sub.3
extract was evaporated and the residue was dissolved in 1.0 ml
CHCl.sub.3: Methanol (1:1) mixture.
[0115] Solvent system: Toluene: ethyl acetate: glacial acetic acid
(12.5: 7.5:0.5)
[0116] Procedure: The test and reference solutions were applied in
two different tracks on a precoated silica gel GF.sub.254 plate
(5.times.15 cm) of uniform thickness (0.2 mm). The plate was
developed in the solvent system to a distance of 12 cm.
[0117] Scanning: The plate was scanned densitometrically at 254 nm
in both the tracks and the fingerprint profiles were recorded.
Visualization of Spots:
[0118] (i) Under UV 254 nm. [0119] (ii) The plate was sprayed with
anisaldehyde-sulfuric acid reagent and heated at 110.degree. C. for
5-10 min. Evaluation: [0120] (i) Under UV 254 nm light (before
spraying):one spot (0.41) was visible in the sample solution track,
corresponding to (Rf 0.41) glycyrrhetic acid of reference track.
[0121] (ii) In daylight (after spraying): Gylcyrrhetic acid was
visible as a dark violet spot in both reference and test solution
tracks.
[0122] The above experiments indicate a sound procedure of
isolating fairly pure glycyrrhizin from the root of the plant
variety `Misree` of CIMAP as well as market available dry root
samples. By this procedure about 0.1 to 0.25% of glycyrrhizin could
be isolated. The protocol can be scaled up or scaled down depending
upon the initial starting material.
Advantages
[0123] 1. The plant extract of Glycyrrhiza glabra and glycyrrhizin
in the Applicant's study facilitates the transport/uptake of
various compounds including antibiotics, antifungal and anticancer
agents, vitamins and amino acids etc. across the membrane thereby
increasing the absorption or bioavailability various biological
systems including bacteria, animal and plant cells. This thus has
immense importance for absorption of the drugs, pharmaceuticals,
nutraceutical and other related compounds and ions by the cells and
hence huge potential of being used as drug/molecule facilitators.
[0124] 2. The present invention enlarges the scope and use of the
natural glycoside glycyrrhizin in therapeutical and nutraceutical
application. [0125] 3. Glycyrrhizin is nontoxic to humans,animals
or plants, is effective at very low concentrations in a
combination,easy to formulate and enhances the uptake/absorption
and activity of drug molecules. Thus the novel use of this compound
as described in the present invention is safe and effective for
bioenhancement of activity of drugs. [0126] 4. Glycyrrhizin by its
presence will not exert any selection pressure for the mutants to
emerge resistant against them and reduce the dosage of antibiotics
or drugs so that their ill effects are reduced.
* * * * *