Filed of the invention The present invention relates to a composition containing extract and/or bioactive fraction/isolate from the plant Zingiber officinale as a bioavailability enhancer. The present invention also relates to a composition containing extract and/or bioactive fraction/isolate from the plant Zingiber officinale with piperine as a bioavailability enhancer.

Background and prior art references Ginger of commerce or'Adrak'is the dried underground stem or rhizome of the zingiberous, herbaceous plant Zingiber officinale Linn, which constitutes one of the five most important major spices of India, standing 3rd or 4th, competing with chillies, depending upon fluctuations in world market prices and world demand and supply positions. Ginger ranks third in value among all the spices exported from India, being next to pepper and cardamom. Indian ginger is further classified as (i) Malabar ginger (Kerala), (ii) Cochin ginger or Wyanad ginger (iii) Himachal ginger, and (iv) Sikkim/N. E. region ginger.

Ginger is valued in medicine as a carminative and stimulant to the gastro- intestinal tract. It is much in vogue as a household remedy for flatulence and colic.

Externally, ginger is used as a local stimulant and rubefacient. It is included among anti- depressants and it forms an ingredient of some anti-narcotic preparations. Besides its stimulant and carminative properties, it is used in toothaches, gout and rheumatism. The essence of ginger is used to promote digestion. Ginger is reported to act powerfully on the mucous membrane. Beneficial results have been obtained when it has been administered in pulmonary and catarrhal affections. Externally, ginger has been used as curative for headaches, paralysis and rheumatism and internally with other ingredients in intermittent fevers (Wealth of India, Raw Material vol XI (1976), 89, PID, CSIR, New Delhi).

Based on our past experience with the development of piperine as bioavailability enhancer from plants which are part of human diet as well as being medicinal, we took up this plant to evaluate its bioavailability enhancing effect, if any, based on its attributes bearing some similarities to the plant sources of piperine.

Several approaches have been adopted in the past to maximize oral bioavailability, such as (a) particle size reduction (micronization, nanonization, etc. ,) (b) polymorphic or crystal size and form selection, (c) solubilization of lesser soluble drugs by way of chemical modifications, complexation and use of co-solvents/surfactants, (d) targeted delivery of drug at the site of action, (e) controlled drug delivery by film coating or use of polymeric matrices for sustained release of drugs, (f) prodrug approach, and (g) microencapsulation using liposomes.

However, based on information from Ayurvedic literatures, a new approach of increasing the bioavailability of drugs including poorly bioavailable drugs had been conceptualized at RRL, Jammu. One of the groups of herbals which has been documented very frequently as essential part of about 70 % of Ayurvedic prescriptions, was noted to be'Trilcatu', that comprises three acrids viz. long pepper, black pepper and dry ginger in equal proportions. A single major alkaloidal constituent from peppers (piperine) was found to be responsible for bioavailability enhancing effect. The role of ginger is to regulate intestinal function to facilitate absorption. Influence of piperine was extensively studied on anti-TB drugs. It was determined that in combination with piperine the dose of rifampicin can be reduced by about 50% while retaining the therapeutic efficacy of this anti-TB drug at par with the standard dose (450 mg). Based on these findings several other reputed plants were evaluated for bioavailability/ bioefficacy enhancing activity. Polar and non-polar extracts of parts of a few plants viz., Cuminu7n cyminum and Carum carvi increased significantly (25-435 %), the bioavailability of a number of classes of drugs, for example, but not limited to, antibiotics, antifungals, anti-virals, anticancer, cardiovascular, CNS, anti- inflammatory/anti-arthritic, anti-TB/antileprosy, anti-histaminic/respiratory disorders, corticosteroids, immunosuppressants, anti-ulcer. Such extracts either in presence or absence of piperine have been found to be highly selective in their bioavailability/ bioefficacy enhancing action.

United States Patent no 5,972, 382 by Majeed, et al titled as"Use of piperine as a bioavailability enhancer"discloses compositions and methods for the improvement of gastrointestinal absorption and systemic utilization of nutrients and nutritional supplements, wherein the compositions comprise a minimum of 98% of pure alkaloid piperine. The method comprises oral, topical, or parenteral administration of the

compositions of the invention. A new process for the extraction and purification of piperine is also disclosed.

US patent 5,536, 506 by Majeed, et al. titled as"Use of piperine to increase the bioavailability of nutritional compounds"discloses a new composition and method for the improvement of gastrointestinal absorption and systemic utilization of nutrients and nutritional supplements, wherein the composition comprises an extract from the fruit of Piper containing a minimum of 98% of pure alkaloid piperine. The method comprises oral, topical, or parenteral administration of the compositions of the invention. A new process for the extraction and purification of piperine is also disclosed.

Another United States Patent 5,744, 161 by Majeed, et al. titled as"Use of piperine as a bioavailability enhancer"discloses compositions and methods for the improvement of gastrointestinal absorption and systemic utilization of nutrients and nutritional supplements, wherein the compositions comprise a minimum of 98% of pure allcaloid piperine. The method comprises oral, topical, or parenteral administration of the compositions of the invention. A new process for the extraction and purification of piperine is also disclosed.

Chemistry of Zingiber officinale On steam distillation, dried, cracked and comminuted ginger yields 1-3% (average 2%) of pale yellow, viscid oil. The oil possesses the aromatic odor but not the pungent flavor (bite) of the spice. Of course, the odor of the oil is quite lasting. The physico-chemical characteristics of the oil vary depending upon the variety, period of storage and method of extraction etc (E. Guenther, The Essential Oils vol v V. D. Van Nostrand Co. , Inc, NY (1952), 116). It is more economical and convenient to recover oil or oleoresin from dried ginger than from fresh ones. However, fresh scrapings of ginger are quite rich in oil and should be immediately used for steam distillation, otherwise 60- 80% of volatile oil is lost. Ginger oleoresin is obtained by extraction of powdered dried ginger with suitable solvents like alcohol, acetone or any other efficient solvent. Unlike volatile oil, it contains both the volatile oil and non-volatile pungent principles for which ginger is so highly valued. Concentration of the solvent extracts under vacuum and on complete removal of even trace of solvent used yields the oleoresin of ginger. The quantitative composition of the oleoresin depends upon the solvent used. Ginger oleoresin (Gingerin) generally contains following types of compounds: Gingerols,

Zingerones, Shogaols, volatile oil, resins, phenols etc. Ginger oleoresin is manufactured in India and abroad and is in great demand by the various food industries.

Chemical composition of ginger oil and oleoresin The oil contains sesquiterpene hydrocarbons (50% or more), sesquiterpene alcohols, monoterpenoids and associated compounds, esters of acetic acid and caprylic acid and a trace of chavicol. The sesquiterpene hydrocarbons present are Zingiberene (oc and (3,-35. 6%), ar-curcumene (17.7%), farnesene (9. 8%) and relatively smaller amounts of P-bisabolene, y-selinene, (3-elemene and P-sesquiphellandrene. Oxygenated sesquiterpene constituents include zingiberol and two other isomeric alcohols.

Monoterpene hydrocarbons present in the oil include camphene, ex and (3-pinene, cumene, myrcene, limonene, p-cymene and ß-phellandrene. The oxygenated monoterpenes and associated compounds present are 2-heptanol, 2-nonanal, n-nonanal, n-decanal, methyl heptenone and 1,8-cineole. (M. C. Nigam, I. C. Nigam, L. Levi & K. L. Handa Can J. Chem 42 (1964), 2610) Studies on the oils derived from ginger from Jamaica, Nigeria, Sierraleone, China, India and Australia have shown that the composition of the oils have similar patterns, although quantitative differences do exist. The content of zingiberenes lies in the range of 20-30%; the ranges of values for a few other constituents are: p-bisabolene, 5-12%; ar-curcumene, 6-19% and P-sesquiphellandrene, 7-12%. The citral (geranial and neral) content is highly variable; the Australian oils are rich in citral (8-27% av. 19. 8% ; a sample prepared from fresh rhizomes by extraction at r. t. contained 8. 8% geranial and 1.5% neral).

The pungent principles of ginger are non-volatile. These can be extracted from coarsely ground-dried spice by using some suitable solvent. They mainly consist of oxymethyl phenols, the major components being gingerol, shogaol, zingerone and paradol. (D. W. Connell and M. D. Sutherland, Aust. J. Chem. Soc. 22 (1969), 1033; E. K. Nelson, J. Am. Chem. Soc. 39 (1917), 1466) The present invention is directed to preparation of active extracts/bioactive fraction/isolate from the plant Zinziber officinale which include their chemical characterization, fingerprint profiling and methods of using such products to enhance bioavailability and/or bioefficacy of drugs, natural products and essential

nutraceuticals. The present invention is directed to preparation of composite bioenhancers comprising polar and non-polar extracts of parts of Zingiber officinale andl or piperine (Ex: Piper nigrum and Piper longum) which increased significantly (25-435 %), the bioavailability of a number of classes of drugs, for example, but not limited to antibiotics, antifungals, anti-virals, anticancer, cardiovascular, CNS, anti- inflammatory/anti-arthritic, anti-TB/antileprosy, anti-histaminic/respiratory disorders, corticosteroids, immunosppressants, anti-ulcer. Such extracts/bioactive fractions of Zinziber officinale either in presence or absence of piperine (Ex : Piper nigrum and Piper longue) have been found to be highly selective in their bioavailability/bioefficacy enhancing action.

There is a great interest and medical need for the improvement of bioavailability of a large number of drugs which are (a) poorly bioavailable, (b) given for long periods, and are (c) toxic and expensive. Maximizing oral bioavailability is therapeutically important because the extent of bioavailability directly influences plasma concentrations and consequently therapeutic efficacy and dose related toxic effects resulting after oral drug administration. Poorly bioavailable drugs remain sub-therapeutic because a major portion of a dose never reaches the plasma or exerts its pharmacological effect unless and until very large doses are given which may lead to serious side effects. Any significant improvement in bioavailability will result in lowering the dose or the dose frequency of that particular drug. Besides, inter-subject variability is inversely correlated with the extent of bioavailability. Therefore, low oral bioavailability leads to high variability and poor control of plasma concentration and pharmacodynamic effects. Inter-subject variability is particularly of concern for a drug with a narrow safety margin.

Incomplete oral bioavailability has various causes. These include poor dissolution or low aqueous solubility, poor intestinal membrane permeation, degradation of the drug in gastric or intestinal fluids and pre-systemic intestinal or hepatic metabolism. The normal practice to offset some of these problems has been to increase the dosage as stated earlier which has the concern toxicity of patients'non-compliance.

Many therapeutic treatments are also accompanied by loss of essential nutraceuticals in the course of therapy. The present invention improves nutritional status by increasing bioavailability/bioefficacy of various nutraceuticals also, which include metals and vitamins. The bioenhancers of the invention also have the potential to enhance the

bioefficacy of a drug without influencing its plasma concentrations for various reasons, some of which, but not limited to, are described later in this invention under Section on 'Bioavailability/Bioenhancing activity' Objects of the invention The main object of the invention is to provide a active of extract and bioactive fraction obtained from Zingiber officinale.

Another object of the invention is to evaluate bioenhancing/bioavailability of Zingiber officinale extract or bioactive fraction in combination with drugs, nutrients, nutraceuticals, micronutrients and herbal drugs/products.

Still another object of the invention is to provide a bioenhancer composition comprising active principles of Zingiber officinale in combination with drugs, nutrients, nutraceuticals, micronutrients and herbal drugs/products.

Still another embodiment of the present invention is to provide a bioenhancer composition comprising extract/isolate and/or bioactive fractions obtained from Zingiber officinale, piperine and one or more selected from the group consisting of drugs, nutrients, nutraceuticals, micronutrients and herbal drugs/products.

Yet, another object of the invention is to provide a process for isolating bioactive faction from Zingiber officinale useful as a bioenhancer.

Yet another object of the invention is to provide a process for isolating bioactive faction from Zingiber officinale using aqueous and/or alcoholic solvent.

Summary of the invention Accordingly, the present invention provides a bioenhancing composition comprising an effective amount of an extract and/or one or more bioactive fractions/ isolates of Zingiber officinale ; one or more additive selected from drugs, nutrients, nutraceuticals, herbal drugs/products, micro nutrients, antioxidants and pharmaceutically acceptable additives/excipients; and optionally an effective amount of pipeline or extract of piper nigrum or piper longue. The invention also provides a process for the preparation of aqueous extract, aqueous alcoholic extract and bioactive fraction from plant Zingiber officinale useful as a bioenhancer/bioavailability facilitator.

Brief description of the accompanying drawings Figure 1 represents flow sheet for preparation of ginger juice, ar. extract and aq. alcoholic extract from plant Zingiber officifzale

Figure 2 represents flow sheet for fractionation of extracts of plant Zingiber officinale.

Figure 3 represents HPLC chromatogram of dry extract (juice) of Zingiber ogoinale Figure 4 represents HPLC chromatogram of aqueous extract of Zingiber officinale Figure 5 represents HPLC chromatogram of aqueous alcoholic extract of Zingiber officinale Figure 6 represents HPLC chromatogram of fraction 1 of aqueous alcoholic extract of Zingiber officinale Figure 7 represents HPLC chromatogram of fraction 2 of aqueous alcoholic extract of Zingiber o, gcmale Figure 8 represents HPLC chromatogram of fraction 3 aqueous alcoholic extract of Zingiber officinale Detailed Description of the Invention Accordingly, the present invention provides a bioenhancing composition comprising: i. an effective amount of an extract and/or one or more bioactive fractions/ isolates of Zingiber officinale ; ii. one or more additive selected from drugs, nutrients, nutraceuticals, herbal drugs/products, micro nutrients, antioxidants and pharmaceutically acceptable additives/excipients; and iii. optionally an effective amount of piperine or extract of piper nigrunZ or piper longue In an embodiment of the invention provides a composition, in which the amount of Zingiber officinale extract used is in the range of about 2.0 to 250 mg Another embodiment provides a composition, wherein the amount of Zingiber ogcinale fraction/pure isolates used is in the range of about 0.5 to 75 mg Still another embodiment provides a composition, wherein the amount of piperine used is in the range of about 5 to 50 mg.

Yet another embodiment, wherein the amount of the extract used is in the range of about 10 to 150 mg.

Yet another embodiment, wherein the amount of the fraction/pure isolates used is in the range of about 0.5 to 45 mg.

Yet another embodiment, wherein the amount of piperine used is in the range of about 3 to 15 mg.

Still another embodiment, the piperine is isolated from piper nigrum, piper longum or its oleoresin.

Another embodiment of the invention provides a composition in which the drugs used are selected from the group consisting of antibiotics, antifungal, antiviral, anticancer, cardiovascular, CNS drugs, anti-inflammatory/anti arthritic, anti- TB/antileprosy drugs, anti histamines/drugs for respiratory disorders, corticosteriods, immuno-suppressants, anti-ulcer drugs and herbal drugs.

Yet another embodiment, the antibiotic used is selected from the group consisting of quinolones, macrolides, cephalosproins, penicillins and aminoglycosides wherein quinolone is selected from the group consisting of Ciprofloxacin, Pefloxacin, Ofloxacin and Norfloxacin; the macrolide is selected from the group consisting of Erythromycin, Roxythromycin and Azithromycin; the cephalosproins is selected from the group consisting of Cefalexin, cefatrioxone, cefixime and Cefadroxil; the penicillin is selected from the group consisting of Amoxycillin and Cloxacillin; and aminoglycocide is selected from the group consisting of Amikacin and Kanamycin.

In yet another embodiment, the anti-fungal drug used is selected from the group consisting of Fluconazole, Amphotericin B and Ketoconazole.

In yet another embodiment, the antiviral drug used is selected from the group consisting of Acyclovir and Zidovudine.

In yet another embodiment, the anticancer drug is selected from the group consisting of Methotrexate, 5-Fluorouracil, Doxorubicin and Cisplatin.

In yet another embodiment, the cardiovascular drug is selected from the group consisting of Amlodipin, Lisinopril, propranolol and Atenolol.

In yet another embodiment, wherein CNS drugs is selected from the group consisting of Alprazolam and Haloperidol In yet another embodiment, wherein anti-inflammatory/anti-arthritic drug is selected from the group consists of Diclofenac, Piroxicam, Nimesulide and Rofecoxib.

In yet another embodiment, anti-TB/anti-leprosy drug is selected from the group consisting of Rifampicin, Ethionamide, Isoniazid, Cycloserine, Pyrazinamide, Ethambutol and Dapsone In yet another embodiment, antihistamine/drugs for respiratory disorders compound is selected from the group consisting of Salbutamol, Theophylline, Bromhexine and Loratidine

In yet another embodiment, corticosteriod is selected from the group consisting of Prednisolone, dexamethasone and Betamethasone In yet another embodiment, immuno-supressant is selected from the group consisting of Cyclosporin A, Tacrolimus and Mycophenolatemofetil.

In yet another embodiment, wherein the anti-ulcer compound is selected from the group consists of Rantidine, Cimetidine and Omerprazole.

In another embodiment, the herbal product/drug is selected from Echinacea, Tinospora cordifolia, Picrorrhiza kurroa, Aegles marmelos, Andrographis paniculata, Emblica ribes, Asparagus racemosus, Terminalia chebula Withania somnifera, Centella asiatica and/or their mixture thereof.

In yet another embodiment, wherein the nutrient is selected from group consists of sugar, carbohydrates, fats and proteins.

One more embodiment of the present invention provides a composition, wherein vitamin used is selected from the group consisting of Vitamin A, Vitamin E, Vitamin B 1, Vitamin B6, Vitamin B 12, Vitamin C and Folic acid.

The antioxidant used for preparing the bioenhancing composition is selected from the group consisting of ß-Carotene, Silymarin, Selenium, Lycopene and Ellagiogallotannins The natural herbal product used is selected from the group consisting of Curcumin, Boswellic acids and Ruti n and essential micro nutrients is selected from the group consisting of Methionine, Lysine, Leucine, Valine, Isoleucine, Zinc, Calcium, Glucose, Potassium, Copper and Iron In yet another embodiment of the invention, the plant extract of Zingiber officinale or its fraction/pure isolate used is extracted from any plant parts of Zingiber officinale One more embodiment of the invention related to administration of the bioenhancing composition. The composition is administered through oral, parenteral, nasal, inhalation including nebulisers, rectal, vaginal, transdermal and any others suitable routes.

In yet another embodiment, the bioenhancing effect of the extracts/fractions/pure isolates of Zingiber officinale alone or in combination with piperine is selective in enhancing the bioavailability/bioefficacy of a drug, nutraceutical, and herbal drug/ formulation.

In another embodiment of the invention, wherein bioavailability/bio-enhancing activity provided by Zingiber officiyaale alone is up to 75 % Yet another embodiment, the composition containing Zingiber offlci7lale alone provides bioavailability/bio-enhancing activity in the range of 30-75 % One more embodiment of the invention provides a composition, wherein the composition containing piperine and Zingiber officinale, further enhances the bioavailability of drugs in the range of 10 to 85% beyond Ziragiber officinale alone.

Another embodiment, the dosage of bioehancer from Zingiber officinale as extract is in the range of 10 to 30-mg/kg/body weight and piperine is in the range of 4 to 12 mg/kg/body weight.

Another embodiment, the dosage of bioehancer from Zingiber officinale as bioactive fraction is in the range of 5 to 15-mg/kg/body weight, preferably 30- mg/kg/body weight, and piperine is in the range of 6 to 10 mg/kg/body weight, preferably 8-mg/kg/body weight.

One more embodiment of the present invention provides a process for the preparation of an aqueous extract, aqueous alcoholic extract and bioactive fraction from the plant Zingiber officinale, said process comprises steps of: a) extracting crushed plant material with water or aqueous alcoholic solvent at a temperature range of 95-100°C ; b) cooling and filtering the extract of step (a) to obtain a clear aqueous extract or aqueous alcoholic extract; c) evaporating the aqueous extract of step (b) under reduced pressure at 60°C to obtain an concentrated aqueous extract; d) freeze drying the concentrated aqueous extract of step (c) to obtain a dried aqueous extract; e) evaporating the solvent from aqueous alcoholic extract of step (b); macerating dried aqueous alocholic extract obtained from step (e) of with chloroform, g) separating the chloroform soluble fraction from step (f) to obtain fraction 1 and an insoluble fraction; h) dissolving the insoluble fraction from step (g) in water and adding alcoholic solvent, stirring and filtering the mixture to separate aqueous alcohol soluble portion and a residue;

i) distilling the aqueous alcohol soluble fraction of step (h) under reduced pressure to obtain fraction 2; and j) dissolving the residue of step (i) in water and freeze drying it to get fraction 3.

Another embodiment of the invention, wherein the solvents used are selected from aqueous, aqueous-alcoholic, ketonic, etheral and halogenated solvents.

Still another embodiment of the invention, wherein the alcoholic solvent used is selected from the group consisting of methanol, ethanol, propanol and/or aqueous alcoholic solvent.

Still another embodiment of the invention, the ratio of plant material used in step (a) to the solvent used is in the range of 1: 1 to 1: 3, preferably 1: 2.5.

Bioavailability/bioefficacy enhancing activity The aqueous, aqueous-alcoholic, ketonic, ethereal, halogenated solvents extracts of the plant parts were evaluated with different therapeutic categories of drugs and nutrients (vital amino acids, metals, antioxidants, vitamins) and herbal drugs. The bioavailability/bioefficacy enhancing (BE) activity of the extracts was found to be consistent from 10 mg to 150 mg irrespective of the amount of the drug (s) present in the formulation. Sub-bioactive fractions of the active extracts were also evaluated, with the same categories of drugs. The BE activity of the bioactive fraction (s) increased corresponding to their proportions in the parent extract. The doses of the fraction (s) responsible for the BE activity ranged from 0.5 to 45 mg. The parent extract as well as the active fraction (s) were found to be active individually as well as in combination with each other with different categories of drugs. The bioenhancer activity of the fraction (s) was found to be consistent from 2.0 mg to 30.0 mg irrespective of the amount of the drug (s) present in the formulation. The BE activity of the fraction (s) was more enhanced as compared to that of the parent extracts.

The extracts or its bioactive fractions were found to be 25-80 % more active when used individually in combination with piperine (1-piperoyl piperidine). Besides both the parent extracts as wells as their bioactive fractions in different combinations showed pronounced activity ranging from 20-70 % in presence of piperine. The amount of piperine in these formulations ranging from 03-15 mg.

The extracts or its fractions either in presence or absence of piperine have been found to be highly selective in their bioavailability enhancing activity. This is apparent from the degree of bioavailibility enhancement caused by these extracts/bioactive fractions. It varies from nil to nearly significant (15 %) to highly significant (120 %).

The reasons for this rather selective pattern as applicable to formulations with or without piperine may be as follows: The extract or its bioactive fraction (s) have been found to be highly selective in their bioavailability enhancing activity. This is more than apparent from the degree of bioavailability enhancement caused by these extract/fraction (s). It varies from almost nearly significant (20%) to highly significant (200%). The reasons for this non-uniform or rather selective pattern as applicable to formulations with or without piperine may be as follows: 1. The extracts/fractions may be enhancing the absorption/transport of certain drugs/nutrients from the gastrointestinal tract.

2. They may be inhibiting partially the specific drug metabolising enzymes, responsible for selective biotransformation of molecules, thus prolonging the elimination or biological half-life of the drug.

3. An increased penetration of therapeutic drugs into their cellular/molecular targets could also be one of the reasons.

4. The formulations may also affect the protein/tissue binding of active drugs, which may be responsible for enhanced bioenhancing effect.

5. Direct potentiation of mechanism of action of a drug may be an important factor contributing to enhanced bioavailability.

6. An enhanced immune response of the host by the incorporation of bioenhancer may cause increase in therapeutic response of the active drugs.

7. A combination of either two or more than two factors as enumerated above [Serial No. 1-6] may be prevalent in the overall bioenhancing effect.

8. The reasons for this selective pattern may be attributable to one or more than one of the following factors: (a) promoting the absorption of drugs from GIT (b) inhibiting/ reducing the rate of biotransformation of drugs in the liver or intestines (c) modifying the immune system in a way that the overall requirement of the drug is reduced substantially (d) increasing the penetration or the entry into the pathogens even where they become persistors within the macrophages such as for Mycobacterium

tuberculosis and such others. This eventually ensures the enhanced killing of these organisms well secured within the places otherwise inaccessible to the active drug, (e) Inhibiting the capability of pathogens or abnormal tissue to reject the drug e. g., efflux mechanisms frequently encountered with anti-malarial, anti-cancer and anti- microbial drugs (f) modifying the signaling process between host and pathogens ensuring increased accessibility of the drugsto the pathogens (g) Enhancing the binding of the drug with the target sites such as receptors, proteins, DNA, RNA and the like in the pathogen, thus potentiating and prolonging its effect leading to enhanced antibiotic activity against pathogens (h) Besides above plausible modes of action the bioenhancer agents may also be useful for promoting the transport of nutrients and the drugs across the blood-brain barrier which could be of immense help in the control of diseases like cerebral infections, epilepsy and other CNS problems.

Primarily, but not exclusively, the invention enhances the carrier-mediated entry of the drug and also thepassive diffusion and the active transport pathways in the tissue which are responsible for transporting physiological substances such as nutraceutical to their target sites. As applicable to any mechanism of action the products of this invention contribute in a synergistic and/or additive manner so that most drugs and nutraceuticals in presence of the products described in the present art are more bioavailable or bioefficaceous as a result of one or more of the mechanisms. The bioavailability and the bioefficacy of drugs and nutraceuticals is also relevant to animal health besides being important for humans. The invention therefore is also intended to be used in veterinary preparations.

The invention further relates to the isolation of an extract and/or its fraction from the plant Zingiber officinale, its standardization with its intended use as drug bioavailability enhancer for the drugs belonging to therapeutic categories such as antimicrobial, antifungal, anti-viral, antitubercular, antileprosy, antiinflammatory, antiarthritic, cardiovascular, antihistaminics, respiratory distree relieving drugs, immunosuppressants, nutraceuticals in compositions to be administered orally/parenterally, topically, inhalations (including nebulizers), rectally, vaginally in human beings and/or veterinary conditions.

In addition, the invention relates to the preparation of a formulation containing extract and/or its fraction/isolate from the plant Zingiber officiiiale, and piperine, its standardization with its intended use as drug bioavailability enhancer for the drugs

belonging to therapeutic categories such as antimicrobial, antifungal, anti-viral, antitubercular, antileprosy, antiinflammatory, antiarthritic, cardiovascular, antihistaminics, respiratory distress relieving drugs, immunosuppressants, nutraceuticals in compositions to be administered orally/parenterally, topically, inhalations (including nebulizers), rectally, vaginally in human beings and/or veterinary conditions The bioavailability enhancer principle may be any extract, its bioactive fraction and/or a pure isolate from the plant.

The bioavailability enhancer principle may be any extract, its bioactive fraction and/or a pure isolate of the plant in combination with piperine A process for the preparation of extract (s) /bioactive fractions (s)/ pure isolate (s) which may involve the use of water, alcohol, combinations of water and alcohol, halogenated hydrocarbons, ketones, ethers as solvents.

A process for the preparation of extract (s) /bioactive fractions (s)/ pure isolate (s) having piperine which may involve the use of water, alcohol, combinations of water and alcohol, halogenated hydrocarbons, ketones, ethers as solvents A process for preparation of bioactive fractions/pure isolates with or without piperine making use of physical techniques like dialysis/molecular sieves/membranes, variety of chromatographic techniques and/or liquid-liquid or solid phase extractions, followed by their complete finger print profiles (HPLC/HPTLC/LC-MS-MS) The formulation of a drug selected from any of the therapeutic categories of the drugs, nutraceuticals, herbal drugs/formulations in combination with the bioenhancer may be intended for routes of administration viz. , oral, parenteral, nasal, inhalation including nebulisers, rectal, vaginal, transdermal and others.

The bioenhancing effect of the extracts/bioactive fractions/pure isolates of Zingiber officinale either alone or in combination with piperine is selective and does not enhance the bioavailability/bioefficacy of each and every drug, nutraceutical, herbal drug/formulation.

The amount of the extracts in the bioavailablity/bioefficacy enhanced formulation (s) may range from 05 to 75 mg irrespective of the amount of drugs in the formulation (s).

The amount of the fraction/pure isolate in the bioavailability/bioefficacy enhanced formulation (s) may range from 1.0 to 30.0 mg irrespective of the amount of drug (s) incorporated in the formulation (s).

That the extracts/fractions/pure isolates or piperine express no biological or toxicological effect of their own at the doses at which they are intended The following examples are intended to demonstrate some of the preferred embodiments and in no way should be construed to limit the scope of the invention. Any person skilled in the art can design more formulations, which may be considered as part of the present invention.

EXAMPLES Example 1 Preparation of colourless, non-pungent piperine by a novel process. Commercially available Piper nigrum or Piper longum or their oleoresins have been used as the source material. 20 kg long pepper oleoresin is extracted with chlorinated solvents like CHC13, CH2C12, C2H4C12 (25 litre) for six hours or 20 kg black pepper powder is soxhletted with toluene for 8 hours. The extracts are concentrated to dryness under reduced pressure and dissolved in ethanol at 78° C. The ethanol solution is adsorbed over neutral A1203 and packed in a glass column. Elution is carried out with CHC13 : EtOH (9: 1) and the eluate is concentrated to dryness and dissolved in minimum quantity of ethanol. The solution is treated with activated charcoal and filtered through a celite bed. The filtrate is concentrated to saturation point, cooled when colorless crystalline precipitate is obtained.

The precipitate is separated by suction filtration and dried.

The specifications of the preferred materials are as under: Colour: Colourless monoclinic prismatic crystals; Melting point: 131° C ; Assay: Minimum (99.1 % (by LC/MS) Example 2 The extract and the active fractions are prepared from the plant material Z. officinalis as per the flow chart accompanying the specification.

Preparation and fully finger printed (HPLC) profile of the products is appended separately.

Example 3 Doses, models/design of experiment and estimation methodology in a typical experiment

Note 1. Bioenhancers (BE) from Zingiber officiyaalis means either the aqueous, or 50% alcoholic extract or fraction No. l.

2. In case of Zingiber officinale, not withstanding the difference in dose of extract or its fraction, the enhancement caused in the bioavailability of the drug with which they are combined remains the same, because the dose of the fraction used is proportionate to its concentration in the extract.

3. The doses remained unchanged even when the bioenhancers (BEs) were used in combination with each other.

Example 4: Doses: (i) Bioenhancer (BE) from Zingiber officinale Extract: 30 mg/kg body weight (Rats) Fraction No 1: 15 mg/kg body weight (Rats) (ii) Piperine: 8 mg/kg body weight (Rats) (iii) Bioenhancer from Zingiber officinale : 35 mg/kg body weight (Rats) As an example of an experiment in Rat (fasted): Drug: Rifampicin, 40 mg/kg BE (Zingiber officinale & others): Doses as in Example No. 4 Experimental procedure: Drug alone/or in combination with BE was administered to rats as per the following design: Group 1: Control Group 2: Rifampicin alone Group 3: BE alone Group 4: Rifampicin + BE (Zingiber officinale) Blood from control/treated animals at predetermined intervals (0-24 hrs) (Total 14 timings) Rifampicin was extracted from the blood (plasma) using dichloromethane. The concentration of rifampicin in the samples was determined using HPLC (Model: Shimadzu 1080 BP); PDA detector; Mobile phase: phosphate buffer: acetonitile (40: 60); Flow rate 1.0 ml/min. Column RP 18.

Control and BE only groups were employed to study the interference of plasma component and the bioenhancer used.

Example 5

The above methodology was adapted for evaluating the bio-enhancing activity of other drugs, micro nutrients, nutracuticals, nutrients and other herbal products and the enhancing effects are tabulated under each heading.

List of drugs, nutraceuticals, herbal formulations cited below as some of the example for the purpose of present invention. The following tables illustrates the bio enhancing activity of Zingiber officinale extract and/or active fraction, Piperine and combination of Piperine + Zingiber officinale extract and/or active fraction.

A. Drugs Categories Drugs Antibiotics Fluoroquinolones Cipro-, Nor-, P-, and 0-floxacins Macrolides Erythro-, Roxythro-, and Azithromycin Cephalosporins Cefalexin, Cefadroxil, cefatrioxone and Cefixime Penicillins Amoxycillin, Cloxacillin Aminoglycosides Amikacin, Kanamycin II. Antifungal Fluconazole, Amphotericin B, Ketoconazole in. Anti-viral Acyclovir, Zidovudine IV. Anti-cancer Methotrexate 5-Fluorouracil Doxorubicin Cisplatin V. Cardiovascular Amlodipin drug Lisinopril Atenolol VI. CNS drugs Alprazolam Haloperidol VI. Anti-Diclofenac inflammatory/Piroxicam, antiarthritic Nimesulide (NSAID) Rofecoxib VII. Anti-TB/Rifampicin Ethionamide Antileprosy drugs Isoniazid Cycloserine Pyrazinamide Ethambutol Dapsone VIII. Anti histamines/Salbutamol respiratory Theophylline disorders Bromhexine, Loratidine IX Corticosteroids Prednisolone, dexamethasone, Betamethasone X. Immuno-Cyclosporin A, Tacrolimus suppressants Mycophenolate mofetil Drugs Category: I. Antibiotics: (a) Fluroquinolones

Drug % Enhancement in bioavailability BE from Piperine as BE Piperine + BE from Zingiber officinale Zingiber officinale Ciprofloxacin 68 55 70 P-floxacin 53 61 69 O-floxacin 49 52 67 Norfloxacin Negligible Negligible Negligible (b) Macrolides Drug % Enhancement in bioavailability BE from Piperine as BE Piperine + BE from Zingiber officinale Zingiber officinale Erythromycin 68 95 105 Roxythromycin 72 110 93 Azithromycin 78 89 85 (c) Cephalosporins Drug % Enhancement in bioavailability BE from Piperine as BE Piperine + BE from Zingiber officinale Zingiber officinale Cefalexin 75 90 85 Cefadroxil 68 70 65 Cefatrioxone Nil Nil Nil Cefixime Nil Nil Nil

(d) Penicillins % Enhancement in bioavailability Drug BE from Piperine as BE Piperine + BE from Zingiber officinale Zingiber officinale Amoxycillin 80 120 90 Cloxacillin 76 87 90 (e) Aminoglycosides: Drug % Enhancement in bioavailability BE from Piperine as BE Pipeline + BE from Zingiber officinale Zingiber officinale Amikacin Nil Negligible Nil Kanamycin 65 72 92

II. Antifungal Drug % Enhancement in bioavailability BE from Piperine as BE Piperine + BE Zingiber officinale from Zingiber officinale Fluconazole 120 87 110 Amphotericin B Nil Negligible negligible Ketoconazole 125 105 100 III. Anti-viral Drug % Enhancement in bioavailability BE from Piperine as BE Piperine + BE from Zingiber officinale Zingiber officinale Acyclovir 8 96 85 Zidovudine 105 140 126

IV. CNS drugs: Drug % Enhancement in bioavailability BE from Piperine as BE Piperine + BE from Zingiber officinale Zin iber officinale Alprazolam 766570 Haloperidol Nil Nil Nil V. Anti-cancer Drug % Enhancement in bioavailability BE from Pipeline as BE Piperine + BE from Zingiber officinale Zingiber officinale . Methotrexate 87 65 49 5-Fluorouracil 110 87 93 Doxorubicin 72 68 75 Cisplatin 56 Negligible 45 VI. Cardiovascular: Drug % Enhancement in bioavailability BE from Piperine as BE Piperine + BE from Zingiber officinale Zingiber officinale Amlodipine 68 43 95 Lisinopril 76 85 100 Atenolol Nil Ne li ible Ne ligible Propranolol 76 84 104 VII. Anti-inflammatory/antiarthritic : Drug % Enhancement in bioavailability BE from Piperine as BE Pipeline + BE from Zingiber officinale Zingiber officinale. Diclofenac 90 120 140 Piroxicam 86 110 134 Nimesulide 144 132 165 Rofecoxib Negligible Negligible Negligible VIII. Anti-TB/Antileprosy drugs:

Drug % Enhancement in bioavailabilit BE from Piperine as BE Piperine + BE from Zingiber officinale Zingiber officinale. Rifampicin 65 45 98 Isoniazid Nil nil Neg ible Pyrazinamide Nil nil Nil Ethambutol Nil Nil Nil Dapsone 46 34 55 Ethionamide 56 45 48 Cycloserine 71 67 70 IX. Anti-histamines/respiratory disorders: Drug % Enhancement in bioavailability BE from Piperine as BE Piperine + BE from Zingiber officinale. Zingiber officinale Salbutamol 78 60 92 Theophylline 76 65 80 Bromhexine 46 67 75 Loratidine Nil Nil Nil X. Corticosteroids: Drug % Enhancement in bioavailability BE from Piperine as BE Piperine + BE from Zingiber officinale Zingiber officinale Prednisolone Nil Nil Nil Dexamethasone 76 66 80 Betamethasone 75 72 70 XI. Immunosuppressants: Drug % Enhancement in bioavailability BE from Piperine as BE Piperine + BE from Zingiber officinale Zingiber officinale Cyclosporin A 116 86 120 Tacrolimus 75 105 117 Mycophenolate Nil Nil Nil Mofeit XII. Anti-ulcer

Drug % Enhancement in bioavailability. BE from Piperine as BE Piperine + BE Zingiber officinal. Ziniber officinale. Ranitidine 147 21 165 Cimetidine 98 Nil 76 Ome razole Nil Nil Nil D. Herbal formulations: Drug % Enhancement in bioavailability/bioefficacy BE fromPiperine as BE Piperine + BE from Zingiber officinale. Zingiber officinale. Echinacea 66 75 82 Tinospora cordifolia 67 85 112 Picrorrhiza kurroa 56 78 87 Aegles marmelos Nil Nil Nil Andrographis paniculata 55 63 70 Emblica ribes 65 Nil 72 Asparagus racemosus 44 58 60 Terminalia chebula Nil Nil Nil Withania somnifera 64 55 48 Centella asiatica Nil Nil Nil Nutraceutical % Enhancement in bioavailability Category Dose BE from Piperine Piperine + BE (mg/kg) Zingiber from Z. officinalis officinalis I Vitamins Vit A 1 30 14 40 Vit E 40 27 nil 25 VitBl 10 nil 16 15 Vit B6 0.5 nil nil nil Vit B 12 0.1 ug nil nil nil Vit C 50 25 nil 24 Folic acid 50 ug 34 nil 35 II Antioxidants B-carotene 15 36 34 55 Silymarin 5 28 13 43 Selenium 2 nil nil nil III Natural herbal Products Curcumin 50 43 33 70 Boswellic acid 50 nil nil nil Rutin 40 21 66 78 IV Essential nutritional components Methionine 20 25 23 45 Lysine 40 15 31 40 Leucine 50 17 25 34 Valine 25 25 26 46 Isoleucine 25 31 18 35 Zinc 0. 1 19 nil 20 Calcium 30 nil negligible nil Glucose 50 nil 29 25 Potassium 25 21 nil 19 Copper 30 nil nil nil Iron 0. 5 nil nil nil