CN100425236C - Use of cuminum cyminum extract and piperine for potentiaion of bioeffectacy of anti infectives - Google Patents

Abstract

The present invention relates to the application of an organism reinforcing agent for reducing the medicine resistance of microorganism strains to anti-infection agents of antibiotic and antifungal agents by strengthening anti-infection activity. The organism reinforcing agent can be used for reducing the medicine resistance of bacteria and yeast, and consequently, the organism reinforcing agent can assist the treatment of certain infection and diseases. Moreover, the organism reinforcing agent reduces and inhibits the concentration of anti-infection agents needed by the growth of microorganism strains.

Description

Use of cumin extract and piperine in the preparation of medicines that strengthen the biological effectiveness of anti-infective agents

The present invention relates to the field of chemotherapeutics, and in particular to formulations as oral pharmaceutical compositions containing bioenhancers that increase the bioefficacy of anti-infective agents, whereby the administration of such anti-infective agents is required The dose and/or frequency of administration is reduced while maintaining the therapeutic effect of standard doses of this drug.

Background of the invention

Many human diseases have their causative origins from pathogenic microorganisms, including bacteria, viruses and fungi. The presence of these pathogenic organisms can lead to sepsis, severe upper and lower respiratory tract infections, CNS infections, meningitis, intraperitoneal infections including the peritoneum, genitourinary tract infections, skin infections, and soft tissue infections and a variety of other infections such as systemic mycoses , Candidiasis, including infections caused by skin fungi. During the past 100 years, remarkable progress has been made in the fight against diseases caused by numerous microbial families with the help of numerous therapeutic agents with different chemical and biological properties as short-term and long-term treatments. These antibiotics include aminoglycosides, penicillins, cephalosporins, macrolides, glycopeptides, fluoroquinolones, tetracyclines, first- and second-line anti-TB drugs, antileprosy agents, antiviral agents, polyenes, Triazole and imidazole antifungals, such as pyrimidine derivatives and combinations of trimethoprim and sulphamethoxazole.

Although these agents are effective against pathogenic microorganisms and fungi and thus can be used in the treatment of disease states associated with the presence of these pathogens, increasing evidence suggests that the use of such agents has certain limitations and raises clinical concerns. Some of the factors that have given rise to these concerns are: (a) the increasing resistance of certain strains of bacteria and fungi to one or more known anti-infective agents and resulting in a reduction in the beneficial effects of usual or standard therapeutic doses, ( b) higher doses required to combat this result in undesired toxic side effects, and (c) high cost of treatment and patient-non-compliance. The emergence of such drug-resistant pathogenic microorganisms is also attributed to the uncontrolled overuse of antibiotics and insufficient use or even insufficient doses and irregular frequency of administration. Prolonged and high-dose therapy is also a serious concern, especially in pregnant women, the elderly and children.

While methods that crystallize the rational use of antibiotics can help slow the problem of microbial resistance, new antibiotics must be discovered to combat those strains that are now resistant to most, if not all, existing antibiotics. Thus, there is an increasing need to identify new antimicrobials that can be used to further complement the physician's arsenal against pathogenic microorganisms.

In another approach, two anti-infective agents are combined in a synergistic manner, ie, one anti-infective acts as a synergistic agent of the other. An example of such a combination is trimethoprim-sulfamethoxazole also known as trimethoprim-sulfamethoxazole or TMP-SMX, which was introduced in 1968 as a broad-spectrum antibiotic. Trimethoprim was specifically developed as a synergist of sulfamethoxazole to have a synergistic effect on the antibacterial and delay the development of bacterial resistance. A 1:5 ratio of trimethoprim to sulfamethoxazole achieves a peak plasma concentration of about 1:20, which is an ideal synergistic serum concentration ratio for most susceptible bacteria (Gutman LT , Pediatr Infect Dis 1984; 3:349-57, Olin BR, Facts and Comparisons, Inc. 1998; 408b-409d, Cockerill FR, Edson RS, Mayo Clin Proc 1991; 66: 1260-9).

The combination may also be an anti-infective agent with another chemical agent which is not itself an anti-infective agent but which in combination with said anti-infective agent enhances the effect of the anti-infective agent. An example of such a combination is amoxicillin + clavulanic acid, more commonly known as Augmentin. Amoxicillin is a penicillin-type antibiotic. It is effective against certain strains of various bacteria such as H. influenzae, N. gonorrhea, Escherichia coli, pneumococcus, streptococcus and staphylococcus. Chemically, the agent is closely related to penicillin and ampicillin. The addition of clavulanic acid to amoxicillin in Libertin enhances the antimicrobial effect of this antibiotic against a variety of other bacteria that are often resistant to amoxicillin. Clavulanic acid is produced by fermentation of Streptomyces clavuligerus. It is a β-lactam structurally related to penicillin and has the effect of inactivating numerous β-lactamases by blocking the active site of the enzyme. Clavulanic acid confers specific resistance to the clinically important plasmid-mediated β-lactamases often responsible for the transfer of resistance to penicillins and cephalosporins.

One of the most notable features associated with traditional Indian medicine and well described in Ayurveda is the use of composition, which provides additive, synergistic and potentiating effects to a medicine when used in combination with other medicines . In Ayurveda there are some natural products which have been used as essential ingredients of many preparations used in the treatment of various diseases. The most prominent of these is "Trikatu", which contains black pepper, long pepper and dried ginger. Detailed and systematic studies have shown that one of the active constituents of pepper, piperine, is a potent bioavailability and/or biopotency enhancer of various pharmaceuticals and nutraceuticals. In earlier patents (Indian Patent No. 172684; Indian Patent No. 172690; Indian Patent No. 176433; and U.S. Patent No. 5439891) it has been disclosed to obtain piperine and piperine-containing preparations including anti-TB antibiotics approach, these anti-TB antibiotics have enhanced bioavailability/bioefficacy at lower active drug doses.

purpose of invention

The main object of the present invention is to provide an oral pharmaceutical composition containing a bioenhancer (bioenhancer) for increasing the biological effectiveness of an anti-infective agent, and thus requires a lower dose and/or reduces the administered dose of this anti-infective agent times while maintaining the therapeutic effect of the standard dose of this drug.

Summary of the invention

The present invention relates to a combination in which piperine and other biopotentiators are used as potentiators when combined with different anti-infective agents using bacteria, viruses and yeasts ex vivo, In vivo infection models were used in mice and guinea pigs. The purpose of the present invention is to overcome and avoid the problems existing in the prior art. The use of the products of the present invention provides a low dosage regimen resulting in an enhanced therapeutic effect compared to standard dosages of the drug alone.

Accordingly, the present invention provides a composition for enhancing the therapeutic effect in the case of a reduced dose of an anti-infective agent against a microbial infection, the composition comprising a mixture of an anti-infective agent and a biological enhancer selected from the group consisting of formula 1 Piperine and 3' of formula 2, 5-dihydroxyflavone 7-O-β-D-galacturin-4'-O-β-D-glucopyranose (3', 5-Dihydroxy flavone7-O-β -D-galacturonide-4'-O-β-D-glucopyranoside) or a mixture thereof.

Formula 1

Formula 2

In one embodiment of the present invention, the anti-infective agent is selected from penicillins including semi-synthetics, cephalosporins, aminoglycosides, glycopeptides, fluoroquinolones, macrolides, tetracyclines , first-line and second-line anti-TB drugs, anti-leprosy drugs, oxazolidelones (oxazolidelones), anti-fungal drugs, anti-viral agents and pyrimidine derivatives-sulfonamide compositions.

In a further embodiment of the invention said antifungal agent is selected from polyenes, imidazoles and triazoles.

In yet another embodiment of the present invention, the antiviral agent is selected from Zidovudines, idouridine, acyclovir and ribavirin.

In another embodiment of the present invention, the 3', 5-dihydroxyflavone 7-O-β-D-galacturon-4'-O-β-D-glucopyranose derived from cumin ( Cuminum cyminum) or subfraction of 3',5-dihydroxyflavone 7-O-β-D-galacturon-4'-O-β-D-glucopyranose in pure form or HPLC fingerprint It is used in the form of fraction.

In another embodiment of the present invention, the concentration of the anti-infective agent is 1/2 to 1/8 of the concentration of the anti-infective agent without the biological enhancer.

In yet another embodiment of the present invention, the composition includes one or more pharmaceutically acceptable additives and excipients.

In another embodiment of the invention, said additive/excipient is selected from nutritional agents comprising protein, carbohydrates, sugar, talc, magnesium stearate, cellulose, calcium carbonate, starch-gel paste, and /or pharmaceutically acceptable carriers, diluents and solvents.

In another embodiment of the present invention, said composition is in the form of oral administration.

In yet another embodiment of the present invention, the ratio of the anti-infective agent to the biological enhancer is 1:1 to 1:5.

In yet another embodiment of the invention, said additive does not contribute to the anti-infective properties of said composition.

The present invention also provides a method for preparing a composition for enhancing therapeutic effect in the case of a reduced dose of the anti-infective agent against microbial infection, said composition comprising a mixture of an anti-infective agent and a biological enhancer, the biological enhancer Piperine selected from formula 1 and 3', 5-dihydroxyflavone 7-O-β-D-galacturon-4'-O-β-D-glucopyranose and mixtures thereof of formula 2, the Methods include physical blending techniques.

Formula 1

Formula 2

In one embodiment of the invention, said physical incorporation technique is selected from dialysis, molecular sieves and membrane methods.

In another embodiment of the present invention, the method of preparation of the bioenhancer comprises the use of water, alcohols, combinations of water and alcohols, hydrocarbons, ketones and ethers.

In one embodiment of the present invention, the anti-infective agent is selected from penicillins including semi-synthetics, cephalosporins, aminoglycosides, glycopeptides, fluoroquinolones, macrolides, tetracyclines , first-line and second-line anti-TB drugs, anti-leprosy drugs, oxazolidinones, anti-fungal drugs, anti-viral agents and pyrimidine derivatives-sulfonamide compositions.

In another embodiment of the present invention, the antifungal agent is selected from polyenes, imidazoles and triazoles.

In yet another embodiment of the present invention, the antiviral agent is selected from the group consisting of zidovudine, iodinidine, acyclovir and ribavirin.

In another embodiment of the present invention, the 3', 5-dihydroxyflavone 7-O-β-D-galacturon-4'-O-β-D-glucopyranose is derived from cumin or The subsite 3',5-dihydroxyflavone 7-O-β-D-galacturon-4'-O-β-D-glucopyranose was used in pure form or in the form of the HPLC fingerprint site.

In another embodiment of the present invention, the concentration of the anti-infective agent is 1/2 to 1/8 of the concentration of the anti-infective agent without the biological enhancer.

In yet another embodiment of the present invention, the composition includes one or more pharmaceutically acceptable additives and excipients.

In another embodiment of the invention, said additive/excipient is selected from nutritional agents comprising protein, carbohydrates, sugar, talc, magnesium stearate, cellulose, calcium carbonate, starch-gel paste, and /or pharmaceutically acceptable carriers, diluents and solvents.

In another embodiment of the present invention, said composition is in the form of oral administration.

In yet another embodiment of the present invention, the ratio of the anti-infective agent to the biological enhancer is 1:1 to 1:5.

In yet another embodiment of the invention, said additive does not contribute to the anti-infective properties of said composition.

Brief description of the drawings

Figure 1 shows the antimicrobial composition of the present invention according to the checker board method.

Figure 2 shows the effect of rifampicin alone and in combination with piperine in an in vivo infection model in mice.

Figure 3 shows the use of rifampicin alone and in combination with 3', 5-dihydroxyflavone 7-O-β-D-galacturon-4'-O-β-D- Effects of combined use of glucopyranose.

Detailed description of the invention

Biopotency/Bioavailability

Research from the inventor's laboratory led to the conceptualization of a "bioenhancer", an agent that is not itself a therapeutic entity, but that, in combination with an active drug, results in an enhancement of the pharmacological action of said drug. Such formulations have been found to increase the bioavailability/bioefficacy of some drugs even when the dose of drug in these formulations is low. Evidence has been obtained for a class of drugs that are (a) poorly bioavailable and/and efficacious, (b) require long-term treatment, and (c) are highly toxic and expensive. For example, Indian Patent No. 172690, No. 176433 and US Patent No. 5744161 disclose such techniques. Further research in the inventor's laboratory proves that this bioenhancer can not only increase the bioavailability of various therapeutic agents through various mechanisms of the following serial numbers (a) to (g), but also enhance the bioefficacy. This leads to a new understanding of the factors and strategies involved in reducing the cellular concentrations of drugs that usually do not reach therapeutic levels, and that the therapeutic strategy enables these drugs to be administered even at lower concentrations compared to standard high doses. The bioavailability and/or biopotency of the active drug is increased. Some of these factors are:

(a) In addition to inhibiting the drug resistance of pathogens and abnormal tissues, increasing the permeability or entry of said active drug to said pathogens, even when the pathogens become persisters.

(b) Chemoresistance is a major problem in drug therapy. Mechanisms for the clinical manifestations of de novo and acquired resistance can arise from changes in any step in the cell-killing pathway. These include drug delivery, drug metabolism, drug targets, cellular repair mechanisms and the ability of cells to recognize harmful toxins or pathogens. The usual mechanism for decreased cellular accumulation of drugs is increased expression of P-glycoprotein (P-gp), a membrane transport agent that effectively removes drugs from these cells. Another limiting factor is the high activity of cytochrome P450 (CYP 450)-dependent proteins. These P-gp and CYP 450 proteins have been shown to regulate the oral bioavailability of most drugs. P-gp is thought to be involved in MDR, which is caused by the expression level of this protein in tumors and after drug treatment.

(c) Altering signal processing to ensure increased accessibility of the drug to the pathogen. A growing body of evidence suggests that calcium signaling pathways play a major role in the therapeutic activity of some drugs, with their efflux through P-gp-independent pathways (Vilpo et al, Haematologica 2000:85:806-813). On the other hand, the cAMP-mediated signaling pathway is associated with changes in membrane fluidity (Friedlander G et al, Biochimica et Biophysica Acta 1990; 1022: 1-7).

(d) Immune intervention through NO production, CMI and/or enhancement of humoral immunity, favorably affects Th1/Th2 balance.

(e) Sensitization of specific receptors such as proteins, DNA, RNA, etc., thereby intensifying and prolonging the effects leading to enhanced antibody activity against pathogens and diseases. Regarding these mechanisms, sufficient experimental evidence has been obtained. For example, piperine has been shown to be able to intercalate deeply into the phospholipids of cell membranes (Ray et al., Ind. J Biochem. Biophys 1999; 36:248-251), regulating the fluidity of said membranes, which can alter the Binding transporter activity. Alterations in total permeability can affect (i) specific ion transport channels, and (ii) also cause substantial movement of lipophilic solutes along paracellular pathways. This membrane alteration has also been observed in the action of some polyene antibiotics (Milhaud J et al, Biochimica et Biophysica Acta, 1988; 943:315-325). However, this change in membrane fluidity induced by piperine, as already stated, is short term, fully reversible but greater than that of any selective substance. If not, severe side effects and toxicity manifested themselves in phase II and III clinical trials of reduced doses of anti-TB pharmaceutical preparations, in which 10 mg of piperine was given daily for 6 months, corresponding to standard piperine-free Dosage of anti-TB drugs. Black pepper containing piperine is almost a part of food all over the world. The average amount of pepper consumed per person translates into a much higher piperine content than that used in the formulation of the present invention.

(f) Mechanisms for enhancing drug action and thereby increasing its potency at low doses, eg inhibition of RNA polymerase transcription leads to enhanced action of rifampicin at less than half the standard dose.

(g) increasing the absorption of the drug and/or inhibiting the biotransformation of the drug, thereby increasing the bioavailability of the drug.

The products of the present invention are drug entities acting through synergistic and additive effects based on novel mechanisms, as a result of one or more of the mechanisms disclosed above, making the drug in the formulation more biologically effective and thus Increases the sensitivity of target cells to anti-infective agents.

Description of formulations containing bioenhancers

"Drug" in the present invention refers to a chemical entity (Chemical entity) that has the ability to affect the pathophysiology of an organism and is used for the prevention and treatment of diseases. Drugs include but are not limited to aminoglycosides, penicillins, cephalosporins and other β-lactam agents, macrolides, glycopeptides, fluoroquinolones, tetracyclines, first-line and second-line antibiotics TB drugs, anti-leprosy drugs, antiviral agents, polyenes, triazoles and imidazoles, and combinations such as pyrimidines, sulphamethoxazole. Drugs can be in native, activated and metabolized forms, including charged, uncharged, hydrophilic, hydrophobic, or zwitterionic species, which are transported by simple diffusion, carrier-mediated transport dependent and independent of energy requirements, Entry via ion- and/or voltage-gated channels.

The "bioenhancer" refers to piperine (Formula 1) or other such molecules, as characterized fractions and/or extracts of chemical entities. Methods to obtain piperine in a chemically characterized form that is more than 98% pure are disclosed in Indian Patent Nos. 172689, 172690, 176433, US Patent No. 5439891 and co-pending US Patent Application No. 60/306917/2001. The method for preparing characterized fractions (including HPLC profiles) and pure chemically characterized molecules (Figure 2) from cumin has been disclosed in co-pending patent application No. NF 515/2001. The ratio of these two bioenhancers to drug can vary from 1% to 50% for the site and from 0.1% to 30% for the purified molecule to obtain the desired reduction in MIC values of the anti-infective agent. The ratio of the drug and the bioenhancer and/or the ratio of the combined bioenhancer is controlled by a dose sufficient to produce an enhanced therapeutic effect as measured by the MIC of the formulation, and the dose is low Dosage when the drug is used alone. Pharmaceutically acceptable carriers are usually inert bulky agents added to enable good incorporation of the ingredients and can be solid or liquid. Inert parts of standard pharmaceutical compositions used in this method are also part of the invention.

Research design

Chessboard method:

This is a common method for evaluating antibiotic compositions in vitro. The "checkerboard" refers to the pattern (of vials or wells of a microtiter plate) formed by the multiple dilutions of the two drugs being tested (Eliopoulos GM, Moellering RC. Antimicrobila Combinations). See: In Laboratory Medicine Antibiotics in Laboratory Medicine (Antibiotics in Laboratory Medicine): United States: Williams&Wilkins). In the research of the present invention, the checkerboard is composed of columns and rows, and each small tube (or small hole) in each column contains the same dose of standard drugs (antibiotics/antifungal/anti-TB/antiviral drugs). The dose of drug is diluted along the x-axis, and each vial (or well) in each row contains the same dose of bioenhancer diluted along the y-axis (Figure 3). As a result, each grid of the checkerboard (which represents a vial/well or plate) contained a unique combination of the standard drug and bioenhancer. The concentration range of the standard drug in the present study was 64 μg/ml-0.03 μg/ml, while the range of the bioenhancer tested was 500 μg/ml-0.2 μg/ml. This checkerboard technique can be performed in liquid or semi-solid (agar) media.

Agar method:

In this method, the agar (Mueller Hinton agar, Middlebrook 7H10 agar) was pressure treated and cooled to 50°C-55°C. The combination of the standard drug and the bioenhancer was added to the agar. Serial 2-fold dilutions of each standard drug and the bioenhancer were prepared in the appropriate solvent. In order to maintain the desired concentration of agar and drug and avoid the effect of solvent, the amount of solvent (containing standard drug and bioenhancer) added to the agar is controlled in a small amount (ie ≤ 5% of the total volume). After the agar plates are primed and allowed to dry, the bacteria to be tested are spread on the surface of the agar using a replicating device designed to deliver a standard inoculum (approximately 10 ⁴ CFU/spot). The plates were incubated at 37°C for 24 hours (3 weeks for M. tuberculosis).

Liquid culture method:

The checkerboard assay described above can also be performed in microtiter plates using liquid culture medium. This method can also be used to study the composition of antibacterial/antifungal/antiviral drugs and bioenhancers.

Inhibition by bioenhancers

All bioenhancers were evaluated for their own inhibitory effect, if present, over a concentration range of 500 μg/ml-0.2 μg/ml (Tables 1, 2 and 3).

Table 1: Effects of piperine on microorganisms

Methicillin-resistant Staphylococcus aureus

++++ No suppression, +++20% suppression, ++50% suppression

Table 2: Effects of cumin fractions on microorganisms

Methicillin-resistant Staphylococcus aureus

++++ No suppression, +++20% suppression, ++50% suppression

Table 3: Effects of pure molecules of cumin on microorganisms

Methicillin-resistant Staphylococcus aureus

++++ No suppression, +++20% suppression, ++50% suppression

Example:

The following examples are intended to illustrate some preferred embodiments and are not to be construed as limiting the scope of the invention. Those skilled in the art can devise more forms, which are also part of the present invention.

The preparation of embodiment 1 colorless, non-irritating 99% pure piperine

It was performed according to the methods disclosed in Indian Patent Nos. 1726891 and 172690 and US Patent No. 5439891 and US Application No. 60/306917/2001, which are incorporated herein by reference.

Example 2 Reduction of the MIC of Rifampicin Against Mycobacterium tuberculosis, Mycobacterium avium and Mycobacterium intracellulare when combined with piperine and cumin fractions

The minimum inhibitory concentration (MIC) of rifampicin alone and in combination with piperine was analyzed for mycobacteria using the methods described in the study design.

A 1/2 reduction in the MIC of rifampicin was observed in combination with piperine and cumin fractions (Table 4a, 4b).

Example 3 Reduced Rifampicin Dose Requirements in Combination with Piperine and Cumin Fraction in a Mouse Model of Systemic Infection

This study was carried out to observe the in vivo response of the combination of rifampicin and piperine. Swiss albino mice were infected intravenously with M. tuberculosis H ₃₇ Rv (10 ⁶ CFU/mouse). Infected mice were divided into groups, each group consisted of 6 mice.

Treatment started 24 hours after infection and continued for 4 weeks, once daily for 5 days in a weekly dosing schedule. After 4 weeks, mice were sacrificed and CFU were counted from lung and kidney. Rifampicin at 20 mg/kg alone was able to reduce log 10 CFU by 2-log order. The same effect was observed for rifampin at 10 mg/kg when combined with 20 mg/kg piperine. The cumin fraction was more effective as rifampicin at a dose of 5 mg/kg produced the same reduction in log 10 CFU (Fig. 4a, 4b).

Table 4a MICs of rifampicin alone and in combination with piperine

Table 4b The MIC of rifampicin used alone and in combination with cumin parts

Example 4 Reduction of the MIC of Ciprofloxacin against Staphylococcus aureus, MRSA and Staphylococcus hemolyticus when combined with piperine

Using the method described in the study design, the minimum inhibitory concentration (MIC) of ciprofloxacin (Ciprofloxacin) alone and in combination with piperine (Piperine) against bacterial species was analyzed. A 1/2 to less than 1/8 decrease in the MIC of ciprofloxacin was observed in combination with piperine (Table 5).

Table 5 MIC of ciprofloxacin used alone and in combination with piperine

Example 5 Reduction of the MIC of fluconazole against Candida albicans, Candida parapsilosis and Candida glabrata when combined with piperine

The minimum inhibitory concentration (MIC) of fluconazole alone and in combination with piperine against fungal species was analyzed using the method described in the study design. A 1/2 to 1/8 reduction in the MIC of fluconazole was observed in combination with piperine (Table 6).

Table 6 The MIC of fluconazole used alone and in combination with piperine (P)

Drugs as some examples of the present invention are listed in the attached table 7 of Example 6

references

1. Gutman LT. The use of TMP-SMX in children: a review of side effects and indications (The use of TMP-SMX in children: a review of adverse reactions and indications). Pediatr Infect Dis 1984; 3 : 349-57.

2. Bushby SRM. Synergy of trimethoprim and sulfonamides: History and current status. See: Antibiotics and Antibiosis in Agriculture, London: Butterworths. 1977;64-81.

3. Olin BR, Ed. Drug Facts and Comparisons. St.Louis, Facts and Comparisons, Inc.; 1998: 408b-409d.

4. Cockerill FR, Edson RS. TMP-SMX. Mayo Clin Proc 1991; 66: 1260-9.

Claims (24)

1. The composition that is used for enhancing therapeutic effect under the situation that the dosage of anti-infective agent of antimicrobial infection reduces, described composition comprises the mixture of anti-infective agent and biological enhancer, and described biological enhancer is selected from formula 2 3 ', 5-dihydroxyflavone 7-O-β-D-galacturon-4'-O-β-D-glucopyranone, or 3', 5-dihydroxyflavone 7-O-β- A mixture of D-galacturin-4'-O-β-D-glucopyranose and piperine of formula 1,

2. The composition as claimed in claim 1, wherein said anti-infective agent is selected from penicillins comprising semi-synthetics, cephalosporins, aminoglycosides, glycopeptides, fluoroquinolones, macrolides Classes, tetracyclines, first-line and second-line anti-TB drugs, anti-leprosy drugs, oxazolidinones, antifungal agents, antiviral agents and pyrimidine derivatives-sulfonamides. 3. The composition of claim 2, wherein the antifungal agent is selected from the group consisting of polyenes, imidazoles and triazoles. 4. The composition of claim 2, wherein the antiviral agent is selected from the group consisting of zidovudine, iodine, acyclovir and ribavirin. 5. The composition of claim 1, wherein the 3', 5-dihydroxyflavone 7-O-β-D-galacturon-4'-O-β-D-glucopyranose is derived from However, the 3',5-dihydroxyflavone 7-O-β-D-galacturon-4'-O-β-D-glucopyranone sub-site is used in pure form or in the form of the HPLC fingerprint site . 6. The composition of claim 1, wherein the concentration of the anti-infective agent is 1/21/8 of the concentration of the anti-infective agent in the absence of the bioenhancer. 7. The composition of claim 1, wherein the composition comprises one or more pharmaceutically acceptable additives and excipients. 8. The composition of claim 7, wherein the additive/excipient is selected from the group consisting of nutritional substances including protein, carbohydrates, sugar, talc, magnesium stearate, cellulose, calcium carbonate, starch-gel paste agents, and/or pharmaceutically acceptable carriers, diluents and solvents. 9. The composition of claim 1, wherein said composition is in a form for oral administration. 10. The composition of claim 1, wherein the ratio of the anti-infective agent to the bioenhancer is 1:1 to 1:5. 11. The composition of claim 7, wherein the additive does not contribute to the anti-infective properties of the composition. 12. A method for the preparation of a composition for enhancing the therapeutic effect of said anti-infective agent against a microbial infection with a reduced dose, said composition comprising a mixture of an anti-infective agent and a biological enhancer selected from 3', 5-dihydroxyflavone 7-O-β-D-galacturin-4'-O-β-D-glucopyranone from formula 2, or 3', 5-dihydroxyflavone 7 from formula 2 - a mixture of O-β-D-galacturin-4'-O-β-D-glucopyranose and piperine of formula 1, said method comprising physical blending techniques, 13. The method of claim 12, wherein the physical incorporation technique is selected from the group consisting of dialysis, molecular sieves and membranes. 14. The method of claim 12, wherein the method of preparation of the bioenhancer comprises the use of water, alcohols, combinations of water and alcohols, hydrocarbons, ketones and ethers. 15. The method as claimed in claim 12, wherein said anti-infective agent is selected from penicillins including semi-synthetics, cephalosporins, aminoglycosides, glycopeptides, fluoroquinolones, macrolides , tetracyclines, first-line and second-line anti-TB drugs, anti-leprosy drugs, oxazolidinones, antifungal drugs, antiviral agents and pyrimidine derivatives-sulfonamides compositions. 16. The method of claim 15, wherein the antifungal agent is selected from the group consisting of polyenes, imidazoles and triazoles. 17. The method of claim 15, wherein the antiviral agent is selected from the group consisting of zidovudine, iodine, acyclovir and ribavirin. 18. The method of claim 12, wherein the 3', 5-dihydroxyflavone 7-O-β-D-galacturon-4'-O-β-D-glucopyranose is derived from cumin The 3',5-dihydroxyflavone 7-O-β-D-galacturon-4'-O-β-D-glucopyranose of the celery or subsite was used in pure form or in the form of the HPLC fingerprint site. 19. The method of claim 12, wherein the concentration of the anti-infective agent is 1/2 to 1/8 of the concentration of the anti-infective agent without the bioenhancer. 20. The method of claim 12, wherein the composition includes one or more pharmaceutically acceptable additives and excipients. 21. The method of claim 20, wherein the additive/excipient is selected from nutritional agents comprising protein, carbohydrates, sugar, talc, magnesium stearate, cellulose, calcium carbonate, starch-gel paste , and/or pharmaceutically acceptable carriers, diluents and solvents. 22. The method of claim 12, wherein the composition is in a form for oral administration. 23. The method of claim 12, wherein the ratio of the anti-infective agent to the bioenhancer is 1:1 to 1:5. 24. The method of claim 20, wherein the additive has no effect on the anti-infective properties of the composition.

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