TWI389700B - A novel standardized composition, method of manufacture and use in the resolution of rna virus infection - Google Patents

Description

Novel standardized composition, manufacturing method and use for solving RNA virus infection

The present invention relates to antiviral agents. The present invention provides antiviral preparations obtained from plants and which have improved immune responses and have been found to be effective against HIV infection, AIDS and influenza viruses and infections.

background

The catechins are polyphenolic plant metabolites belonging to the flavonoid family. The molecular formula and molecular weight of catechins are C ₁₅ H ₁₄ O ₆ and 290 g/mol. The catechins and epicatechins are epimers, of which (-)-epicatechin and (+)-catechin are the most common optical isomers found in nature.

Proanthocyanidins or condensed tannins are flavonoid oligomers, and the building blocks are (+)-catechin and (-)-epicatechin. It is the oligomeric end product of the flavonoid biosynthetic pathway and has now been identified and recognized for its beneficial effects on humans. It is abundantly found in fruits, bark, leaves and seeds in the plant kingdom and provides protection against light, oxidation and predators. Proanthocyanidins found in many plants, mainly apple, pine bark, cinnamon bark, litchi peel, peanuts, grape seed, cocoa, grape skin, mulberry, cranberry, black currant, green tea and black tea .

Proanthocyanidins are classified as type A, type B or type C polyphenols based on linkages between consecutive monomer units.

Typically, the bond between successive monomer units of proanthocyanidins is between the fourth position of the "upper" unit and the eighth position of the "lower" unit, producing a type B proanthocyanidin. Alternatively, the linkage may be present between C "upper" _{C. 4} and the lower unit of the unit _6, to produce C type proanthocyanidins. Type B and C polyphenols are abundantly found in many plant sources. When an ether linkage between the successive monomeric units by the C2 "upper" and the cell C4 and the lower unit of the C7 and C ⁶ / C ⁸ bit (respectively) of the oxygen-linked, form A-type proanthocyanidins. Compared with type B and type C polyphenols, type A proanthocyanidins are extremely rare.

Immune response to antigen

The immune system is a collection of mechanisms by which the host protects the host against disease by recognizing and eliminating pathogens. The system's response to pathogens protects the host by identifying foreign proteins and ultimately destroying the source of the additional protein. Even the identification of simple proteins from unicellular organisms involves a series of complex steps that result in the ultimate elimination of organisms from the host. This entire process is an immune response to the presence of foreign proteins or antigens. The infection is resolved by the immune system as an immune response to the antigen, and it can be divided into three phases: activation and movement: white blood cells (WBC) are activated upon recognition of foreign molecules or antigens. Immune cells, such as macrophages and T cells, release substances that attract other immune cells to the location of the identified foreign molecules and thus move countless immune cells to destroy the pathogen.

Regulation: The immune response elicited must be controlled to avoid undue damage to the host. Regulator T lymphocytes assist in the control of immune responses by secreting cytokines that act as messengers of the immune system, thereby modulating excessive immune responses.

Solution: Infection resolution involves binding pathogens and eliminating them from the body. After elimination of pathogens, most of the WBC is destroyed, and the remaining WBCs are called "memory cells" and protect the host against future infections caused by the same pathogen by triggering an early immune response to the pathogen.

Pathogens successfully cause infection when the host is unable to provide protection that is strong enough to eliminate pathogens. In such cases, the antibody produced by the host is not sufficient to neutralize the existing number of antigens. Therefore, the free antigen successfully infects the host. In such cases, supplementation with, for example, antibiotics and antiviral agents is used to reduce the number of antigens. Once the number of antigens is reduced, the immune response is sufficient to eliminate the pathogen.

HIV infection and AIDS:

Human immunodeficiency virus (HIV) is a retrovirus that destroys the immune system. This infection can eventually lead to acquired immunodeficiency syndrome (AIDS), a serious and life-threatening condition that prevents the immune system from functioning properly. HIV primarily infects specific cells in the human immune system: "helper" T lymphocytes (specifically CD4 ⁺ T cells), macrophages, and dendritic cells. When the number of CD4 ⁺ T cells falls below a critical level, cell-mediated immunity is lost and the body is increasingly becoming more susceptible to opportunistic infections.

HIV life cycle: Once HIV enters the host, HIV needs specific host cells to aid in its replication and reproduction. The host cell in the case of HIV is a T cell or a CD4 cell.

1. Identification and binding of the host: HIV searches for CD4 cells and attaches to CD4 cells via a co-receptor on the cell surface by a "lock and key" system. Proteins on the surface of HIV attach to complementary proteins on CD4 cells.

2. Attachment and entry into the host: After attachment, HIV injects viral proteins into the cytosol (cytoplasm) of T cells. This allows the cell membrane to fuse with the outer membrane of HIV.

3. Decomposition of viral proteins: In order to replicate using their genetic material (RNA), it is necessary to dissolve the protective coating surrounding the RNA. Without this step, RNA cannot be converted to DNA (a structural unit of the new HIV replica) and replication ceases.

4. Reverse transcription: Once in the cell, the single RNA of HIV must be converted to double stranded DNA. This step is caused by the enzyme reverse transcriptase. Reverse transcriptases use structural units derived from T cells to help convert viral RNA into DNA. DNA contains the genetic information needed for HIV replication.

5. Replication and assembly into new virions: For replication, newly formed viral DNA must be integrated into the host cell nucleus. This process is not fully understood, but it is believed that this process is aided by viral transport proteins. Upon integration, the virus develops gradually, and the host cell prepares the protein it needs to complete replication. Once the material is available, it is lysed by the virus based on demand and structure and then assembled into new HIV. This process is aided by protease enzymes.

6. Budding from host cells: The final step in the viral replication cycle is called budding. With the collection of genetic material and new extracellular forms formed by the cell membrane of the host CD4 cells, the newly formed HIV breaks and enters the circulation, ready to begin the entire process again.

Current interventions: Current methods of disrupting HIV replication and reproduction include: viral entry inhibitors; membrane fusion inhibitors; and reverse transcriptase inhibitors; integrase inhibitors; protease inhibitors; The FDA has approved many drugs for the treatment of HIV infection. Most of these drugs work by their antiretroviral (ARV) mechanism of action.

Human immunodeficiency virus (HIV) infection poses political, economic, public health, social and scientific challenges to countries around the world. At the end of 2007, an estimated 33.2 million people worldwide lived with HIV/AIDS. Therefore, there is an urgent need to manage and/or treat this disease with safer and more effective drugs. Another challenge presented by this virus is its sensitivity to mutations. The viral proteins of HIV are prone to mutations and thus drug-resistant strains pose additional threats, requiring new classes of drugs.

Influenza virus:

Influenza is an infectious disease caused by the RNA virus (influenza virus) of Orthomyxoviridae, which affects birds and mammals. The infection caused by this virus mainly affects the nose, throat, bronchus and occasionally affects the lungs.

The structure of the influenza virus: influenza viruses are classified into three categories: influenza viruses A, B, and C. The three subtypes of the influenza virus have a very similar overall structure. The virus consists of a viral envelope containing two major types of glycoproteins that wrap around the central nucleus. The central nucleus contains the viral RNA genome and other viral proteins that package and protect this RNA. Hemagglutinin (HA) and neuraminidase (NA) are two large glycoproteins outside the virion.

Influenza A: Type A virus is the most pathogenic human pathogen of the three influenza types and causes the most serious diseases. Type A influenza viruses can be subdivided into different serotypes based on antibody responses to such viruses. The serotypes that have been confirmed in humans are (sorted by the number of known human mass deaths): H1N1, H2N2, H3N2, H5N1, H7N7, H1N2, H9N2, H7N2, H7N3, H10N7. Type A influenza virus has caused several epidemics in the last century and continues to cause annual epidemics. The emergence of new strains of influenza continues to challenge public health and scientific communities. The H1N1 virus is a serotype of influenza A virus and is one of the most virulent strains known to affect humans. The H1N1 serotype caused millions of deaths (Spanish Flu) in 1918 and has recently caused a global pandemic of swine flu.

Influenza virus life cycle: The influenza virus replication and reproduction process is outlined below:

1. Identification and binding of the host : The binding of the virus to the host cell uses the binding of the HA protein to the sialic acid bound to the sugar on the surface of the epithelial cell. Epithelial cells are typically found in the nose, throat and lungs of mammals and in the intestines of birds.

2. Attachment and entry into the host: After binding, the HA protein is cleaved and the virus enters the cell by pinocytosis.

3. Decomposition of viral proteins: Once the virus enters the cell, the pH and environmental conditions of the endosome cause a. A part of HA fuses the viral envelope with the tonoplast, b. M2 ion channel allows protons to enter the viral core, proton acidification The core of the virus causes viral core breakdown and subsequent release of viral RNA and core proteins into the host cytoplasm.

4. Reverse transcription: Viral RNA and core proteins are now transported into the nucleus where RNA is transcribed and further translated into viral proteins.

5. Germination from host cells: HA and NA proteins form clusters near the cell membrane, followed by viral RNA and core proteins, followed by "germination" and propagation of the virus for subsequent infection.

As can be seen from the infection and propagation steps detailed above, HA and NA play an important role in infection. The NA also cleaves sialic acid to prevent HA binding to sialic acid prior to release of the virions.

Current involvement in influenza A virus: There are two types of drugs approved by the US FDA for influenza A virus: ion channel inhibitors such as adamantane (amantadine and rimantadine); Aminase inhibitors such as Oseltamivir (TAMIFLU) and Zanamivir (RELENZA).

Type A influenza virus is susceptible to mutation. Such mutations are primarily viral proteins such as NA, HA and M2 ion channel proteins, and thus inhibitors of such proteins will be ineffective against mutant strains. The potential for mutations and the global global pandemic of influenza A in 2009 indicate the urgent need to provide treatments for the treatment and prevention options of this virus.

R Ichard Anderson et al., Isolation and characterization of polyphenols Type A polymers from cinnamon with insulin-like biolosical activity , in Journal of Agricultural and Food Chemistry, 2004, p. 52, pp. 65-70.

This document describes an aqueous extract of commercial cinnamon and has identified polyphenolic polymers that increase glucose metabolism in cell lines in vitro by about 20 fold. It was prepared using Cinnamomum cassia ; Korintji cassia. This variant has a high content of coumarin and cinnamaldehyde.

This document further describes preparative HPLC methods for the preparation and characterization of such aqueous extracts.

This publication describes the type A double bond of the catechin before the anthocyanin. This document has identified terpolymers (molecular weight 864), tetramers (molecular weight 1152) and oligomers of catechins isolated from cinnamon.

Kilkuskie et al., "HIV and reverse transcriptase inhibitio n by tannins ", in Bioorganic and Medicinal Chemistry Letters, 1992, Volume 2, pages 1529-1534.

This publication evaluates the anti-HIV activity of tannins and condensed tannins and their ability to inhibit reverse transcriptase enzymes. Although this study found some tannins with anti-HIV activity, it also has associated toxicity. This publication discusses three compounds which are condensed forms of catechins. Molecules 40, 44 and 45 are dimers, trimers and tetramers of catechins. This document considers that there is no correlation between the inhibition of RT enzymes and the anti-HIV effects of these tannins. In addition, molecules 44 and 45 exhibited 90% and 73% anti-HIV activity, respectively, but did not exhibit significant RT enzyme inhibition.

Michael Ovadia et al., Patent Application US 2006 275515A1

This patent entitled " Anti-viral preparations obtained from a natural cinnamon extract " describes a natural aqueous extract obtained from cinnamon having antiviral properties. This document describes an aqueous extract of cinnamon which is subjected to salting out precipitation with a salt. This precipitate was redissolved in water or buffer and purified using agarose gel chromatography followed by elution with another buffer and galactose.

A commonly used salting out process refers to the selection of high molecular weight molecules, usually peptides. Therefore, it is apparent from this process that the process described in this document aims to recover high molecular weight molecules (about 10 Kda).

The active ingredient of the composition according to the scope of the patent application has a molecular weight greater than 10 KDA and its response to absorbance at 280 nm is between about 15 and 20 OD. The compound was finally eluted from the sepharon column using phosphate buffer and galactose. Therefore, the final compound will have a high concentration of phosphate and galactose.

Pre-absorption into red blood cells and weight gain, and HIV syncytology studies in influenza A PR 8 virus, parainfluenza (Sendai) virus, in mice infected with influenza or Sendai virus The high molecular weight compound described in this application was tested.

Example 13 of this patent application describes testing with this extract in MT2 cells to examine the effect on syncytia formation. According to Figure 15 of this application, it inhibits syncytia formation at a concentration of 60 to 100 micrograms. Syncytial formation is not a confirmation test for antiviral activity. This is clearly stated in the following publication [Gueseppe pantaleo et al., Eur J immunology 1991, 21, 1771:1774 'dissociation between syncytia formation and HIV spreading. Suppressing Syncytia formation does not have reflect inhibition of HIV infection'].

Although the disclosed extracts demonstrate the ability to inhibit syncytia formation, it should be noted that only some HIV strains cause syncytia formation. Furthermore, syncytia formation cannot be correlated with the presence or progression of HIV infection or AIDS. Syncytial formation is only a phenotype that can be expressed by some strains. Lack of syncytia formation cannot be associated with the absence of HIV or management of infection.

The object of the invention

A first object of the present invention is to provide a composition comprising pentameric proanthocyanidin flavonoids, terpolymers and tetramers from plant sources.

A second object of the present invention is to provide a process for the preparation of a composition comprising pentameric proanthocyanidin flavonoids, terpolymers and tetramers from plant sources such as Cinnamomum , Litchi and Peanut.

A third object of the present invention is to provide a composition that improves the immune response in an individual and is also effective against HIV infection, AIDS and influenza virus and infection.

Statement of the invention

Accordingly, the present invention is directed to compositions comprising pentameric proanthocyanidin flavonoids ranging from about 0.5% w/w to about 99% w/w, each in the range of from about 0.5% w/w to about 35% w Terpolymers and tetramers in the range of /w, and optionally medically acceptable excipients; preparation of pentamers in the range of from about 55% w/w to about 99% w/w A method for anthocyanin flavonoids, a composition of terpolymers and tetramers each in a concentration range of from about 0.5% w/w to about 35% w/w, the method comprising the steps of: extracting and pulverizing using an organic solvent Plant body to remove toxic substances; drying the plant body to remove the organic solvent; extracting the dried plant body with an aqueous solvent to obtain an extract; and purifying the extract through a chromatography column, followed by concentration, purification, standardization and Drying to obtain a composition; a method of modifying an immune response in an individual in need thereof, the method comprising the step of administering to the individual a medically effective amount of a composition comprising from about 55% w/w to about 99% w/ a pentameric proanthocyanidin flavonoid in the concentration range of w, a terpolymer each in a concentration ranging from about 0.5% w/w to about 35% w/w a tetramer, and optionally a medically acceptable excipient; and a method of treating, preventing, and managing a retroviral infection in an individual in need thereof, the method comprising administering to the individual a medically effective amount of the composition In the step, the composition comprises a pentameric proanthocyanidin flavonoid ranging from about 55% w/w to about 99% w/w, each in a concentration range of from about 0.5% w/w to about 35% w/w. Terpolymers and tetramers, and optionally medically acceptable excipients.

The present invention relates to pentameric proanthocyanidin flavonoids ranging from about 55% w/w to about 99% w/w, each in a concentration range of from about 0.5% w/w to about 35% w/w. Compositions of trimers and tetramers, and optionally medically acceptable excipients.

In one embodiment of the invention, the composition is obtained from a plant source selected from the group consisting of Cinnamon, Litchi, and Peanut.

In another embodiment of the invention, the preferred concentration of the pentameric proanthocyanidin flavonoid is in the range of from about 80% w/w to about 99% w/w, and the trimer and tetramer are each about 0.5. % w/w to a concentration range of about 20% w/w.

In another embodiment of the invention, the pentameric proanthocyanidin flavonoid has a molecular weight of about 1440.

In another embodiment of the invention, the pentamer is a type A proanthocyanidin pentapolymer.

In another embodiment of the present invention, the excipients are selected from the group consisting of gums, granulating agents, binders, lubricants, disintegrating agents, sweeteners, coloring agents, flavoring agents, coating agents, and growth agents. A group of plasticizers, preservatives, suspending agents, emulsifiers, antistatic agents, and spheronization agents.

In another embodiment of the present invention, the composition is formulated to be selected from the group consisting of a tablet, a sugar-coated tablet, a buccal tablet, an aqueous or oily suspension, a dispersible powder or granule, a hard gel capsule or a soft gel. Various dosage forms of the group of emulsions, syrups and elixirs in capsules.

The present invention relates to the preparation of pentameric proanthocyanidin flavonoids ranging from about 55% w/w to about 99% w/w, each in the range of from about 0.5% w/w to about 35% w/w. a method of composition of a trimer and a tetramer, the method comprising the steps of: extracting a pulverized plant body using an organic solvent to remove toxic substances; drying the plant body to remove the organic solvent; and extracting the organic solvent using the aqueous solvent The dried plant body is subjected to an extract; and the extract is purified via a chromatography column, followed by concentration, purification, standardization, and drying to obtain a composition.

In one embodiment of the invention, the pulverized plant system is selected from the group consisting of plants of the genus Cinnamon, Litchi, and Peanut.

In another embodiment of the invention, the organic solvent is selected from the group consisting of ethyl acetate, butyl acetate, amyl acetate, 2-ethylhexyl acetate, and any combination thereof.

In another embodiment of the invention, the extraction is carried out for a period of from about 8 hours to about 12 hours, preferably about 10 hours.

In another embodiment of the invention, the toxic substance comprises coumarin and an aldehyde.

In another embodiment of the invention, the extract is filtered through a two-stage chromatography column.

In another embodiment of the invention, the chromatography column is selected from the group consisting of XAD-1180, XAD-7HP, and XAD-1140 resins.

In another embodiment of the invention, the re-extraction using an aqueous solvent is carried out at a pH in the range of from about 3.8 to about 5.8, preferably at a pH of about 4.0.

In another embodiment of the invention, the re-extraction is carried out at a temperature in the range of from about 30 ° C to 90 ° C, preferably in the range of from about 31 ° C to 40 ° C, for a period of from about 8 hours to about 12 hours, It is preferably about 10 hours.

In another embodiment of the invention, the aqueous solvent is acidified deionized water.

In another embodiment of the present invention, the composition further comprises an excipient selected from the group consisting of a gum, a granulating agent, a binder, a lubricant, a disintegrant, a sweetener, and a colorant. Groups of flavoring agents, coating agents, plasticizers, preservatives, suspending agents, emulsifiers, antistatic agents, and spheronizing agents.

The present invention relates to a method for improving an immune response in an individual in need thereof, the method comprising the step of administering to the individual a medically effective amount of a composition comprising from about 55% w/w to about 99% w/w concentration a range of pentameric proanthocyanidin flavonoids, trimers and tetramers each ranging from about 0.5% w/w to about 35% w/w, and optionally medically acceptable Shape agent.

In another embodiment of the invention, the immune response is improved in a disease selected from the group consisting of, but not limited to, influenza, HIV infection, and AIDS.

In another embodiment of the invention, the immune response is improved in an individual in need thereof.

In another embodiment of the invention, the medically effective amount of the composition ranges from about 1 mg/kg of the individual body weight to about 100 mg/kg of the individual body weight.

In another embodiment of the invention, the method is for treating, preventing, and managing an infection caused by a pathogen in an individual.

In another embodiment of the invention, the pathogen comprises influenza A virus and HIV virus.

In another embodiment of the invention, the viral types are H1N1, H3N2, X4 and R5 tropic viruses.

In another embodiment of the invention, the individual is an animal or a human.

The present invention relates to a method of treating, preventing and managing a viral infection in an individual in need thereof, wherein the method comprises the step of administering to the individual a medically effective amount of a composition as an antiviral agent, the composition comprising about 55% w /w to about 99% w/w concentration range of pentameric proanthocyanidin flavonoids, each of the trimer and tetramer in a concentration range of about 0.5% w/w to about 35% w/w, and Medically acceptable excipients, as appropriate.

In another embodiment of the invention, the composition inhibits influenza A virus, X4 tropism of HIV, and R5 tropic virus.

In another embodiment of the invention, the medically effective amount of the composition ranges from about 1 mg/kg of the individual body weight to about 100 mg/kg of the individual body weight.

In another embodiment of the invention, the individual is an animal or a human.

The present invention relates to a method of treating, preventing and managing a retroviral infection in an individual in need thereof, the method comprising the step of administering to the individual a medically effective amount of a composition comprising from about 55% w/w to about Pentameric proanthocyanidin flavonoids in a concentration range of 99% w/w, trimers and tetramers each in a concentration range of from about 0.5% w/w to about 35% w/w, and optionally Medically acceptable excipients.

In one embodiment of the invention, the retroviral infection comprises influenza A infection and HIV infection and AIDS.

In another embodiment of the invention, the medically effective amount of the composition ranges from about 1 mg/kg of the individual body weight to about 100 mg/kg of the individual body weight.

In another embodiment of the invention, the individual is an animal or a human.

The present invention relates to novel standardized compositions derived from vegetal sources which are standardized as 50% to 99% of flavanoid type A proanthocyanidin pentamers as shown in FIG. The invention is also directed to a method of obtaining a novel standardized composition derived from a botanical therapeutic drug source which is standardized to 50% to 99% of a flavanoid type A proanthocyanidin pentapolymer. The invention also relates to the use of a novel standardized composition derived from a vegetable source, which is standardized as 50% to 99% of a flavanoid type A proanthocyanidin pentapolymer for use in prevention and treatment. And managing HIV and influenza infections.

The invention also relates to the use of a novel standardized composition derived from a vegetable source, which is standardized to 50% to 99% of a flavanoid type A proanthocyanidin pentapolymer, which is used to cause a need Improved immune response to antigen in an individual.

In another embodiment of the invention, the immune response can be treated, managed or prevented intrinsically.

In one embodiment of the invention, the botanical source used to obtain the composition is Cinnamon, Litchi, and Peanut.

In one embodiment of the invention, the novel standardized composition derived from a vegetable source is standardized as a flavanoid type A proanthocyanidin pentamer.

In another embodiment of the invention, as shown in Figure 1, the pentamer has a molecular weight of 1440.

In another embodiment of the invention, the composition comprises a pentamer in the range of 50% to 99%.

In another embodiment of the invention, the composition comprises trimers and tetramers in the range of from 1% to 35%.

In another embodiment of the invention, the composition is characterized by the chromatogram of Figure 5.

In another embodiment of the invention, the monomeric unit of the novel composition is selected from the group of catechins, preferably catechin or epicatechin.

The invention is also directed to a method of making novel compositions by the processes described in this document.

In one embodiment of the invention, the standardized composition comprises a medically acceptable excipient as appropriate.

In another embodiment of the present invention, the excipient is selected from the group consisting of an additive, a gum, a sweetener, a coating agent, a binder, a disintegrant, a lubricant, a disintegrant, a suspending agent, a solvent, and a colorant. Groups of slip agents, anti-adhesives, antistatic agents, surfactants, plasticizers, emulsifiers, perfumes, viscosity enhancers and antioxidants.

In still another embodiment of the present invention, the composition is formulated into, for example, a liquid, a powder, a capsule, a tablet, an injection, a patch, an ointment, a gel, an emulsion, a cream, a lotion, a tooth powder, a spray, and a drop. Formulation. In still another embodiment of the invention, the composition is a powder or a liquid.

The present invention is also directed to a method of obtaining a novel standardized composition derived from a vegetable source, which is standardized to 50% to 99% of a flavanoid type A proanthocyanidin pentamer, wherein the method comprises the following steps :

1. Grinding the vegetable material to a predetermined size;

2. Extract with an organic solvent to remove undesired toxic substances;

3. Aqueous extraction of vegetable powder with deionized water;

4. Purification of the extract using a two-stage chromatography purification unit;

5. Drying, blending and screening to obtain a composition comprising a flavanoid pentamer as shown in Figure 1 in a purity of 50% to 99%.

The invention also relates to the use of a novel composition comprising an excipient as appropriate in the context of the invention for the manufacture of a medicament for the treatment and management of HIV and for the prevention, treatment and management of influenza virus infection. The invention also relates to the use of a composition comprising an excipient as appropriate in the context of the invention for the manufacture of a medicament for the treatment and management of HIV infection and for the prevention, treatment and management of influenza infection in an individual in need thereof Product.

The invention also relates to the use of a composition comprising an excipient as appropriate in the context of the invention for the manufacture of a medicament for the improvement of an immune response in an individual in need thereof. The invention also relates to the use of the compositions of the invention for inducing an improved immune response in an individual in need thereof.

In yet another embodiment of the invention, the individual is an animal and a human.

The present invention is also directed to a method of making a novel standardized composition derived from a vegetable source, which is standardized to 50% to 99% of a flavanoid type A proanthocyanidin pentapolymer, the method comprising the steps of:

1. Grinding plant cinnamon or litchi peel or crushed nut shell with red seed coat;

2. Using an organic (preferably ester) solvent extraction material consisting essentially of ethyl acetate, butyl acetate, amyl acetate or 2-ethylhexyl acetate as a single solvent or a mixture of the above solvents to remove fat And toxins and other aromatic compounds. This step is optional for cinnamon;

3. drying the extracted plant material to remove the solvent;

4. Extraction is carried out with deionized water at pH 4 or at a pH between 3.8 and 5.8, preferably at pH 4.0. Extract purification using two-stage chromatography, one stage for polar molecules and one stage for non-polar molecules;

5. eluting the adsorbed substance with an alcohol solvent;

6. Concentrate the eluted solvent to a fine powder;

7. Dilute the concentrated material with water and spray dry as appropriate to eliminate residual solvent.

The novel composition obtained by the above process comprises 50%-99% pentamer, 1%-35% trimer and 1%-35% tetramer and is characterized as shown in FIG.

The invention is further illustrated in detail by means of the following examples. However, the examples are not to be considered as limiting the scope of the invention.

Example 1

The powdered cinnamon powder having an average size of 1000 gm in the 16 mesh range was soaked in 3000 ml of ethyl acetate and poured into an extractor having a 200 mesh mesh bottom sieve. The bottom stripping agent was recirculated over a period of about 8 hours via a packed mass to achieve an effective extraction. The eluent was discarded and the material was removed from the extractor and dried in a forced air oven at 30 °C. After the solvent is removed by drying, the material is reloaded into the extractor. The packaged material was extracted with 5000 ml of acidified deionized water at pH 4.0 and the extract was recirculated through the bed at 35 ° C for about 8 hours to achieve an effective extraction.

The extract was filtered through a two-stage chromatography column to obtain a composition having 80% of a flavanoid type A proanthocyanidin pentamer (molecular weight: 1440), and the extract was passed through the first column to extract the composition. The relatively low polarity molecules and the second stage chromatographic separation are used for relatively high polarity molecules in the composition. The resins used were equivalents of XAD-1180 and XAD-7HP resins, respectively. The column was thoroughly washed with deionized water to make it free of adhesive material and the eluent was neutral. The column was further eluted with 175 ml of pure isopropanol and the collected eluent was concentrated under vacuum at 40 ° C and diluted with water and spray dried under the following conditions:

Spray dryer: cocurrent flow

Inlet temperature: 140 ° C

Outlet temperature 60 ° C

Nebulizer RPM 14000

The final weight is 5 gm.

Example 2

The powdered cinnamon powder having an average size of 1000 gm in the 16 mesh range was soaked in 3000 ml of ethyl acetate and poured into an extractor having a 200 mesh mesh bottom sieve. The bottom stripping agent was recirculated via the packaged material over a period of 10 hours to achieve an effective extraction. The eluent was discarded and the material was removed from the extractor and dried in a forced air oven at 30 °C. After the solvent is removed by drying, the material is reloaded into the extractor. The packaged material was extracted with 5 liters of acidified deionized water at pH 4.0 and the extract was recirculated through the bed at 35 °C for about 8 hours to achieve an effective extraction.

The extract was filtered through a two-stage chromatography column to obtain a composition of 75% flavanoid type A proanthocyanidin pentamer (molecular weight: 1440). The extract is first passed through a first column to extract relatively low polarity molecules in the composition and the second stage chromatographic separation is used in relatively high polarity molecules in the composition. The resins used were equivalents of XAD-1180 and XAD-7HP resins, respectively.

The column was thoroughly washed with deionized water to make it free of adhesive material and the eluent was neutral. The column was again stripped with 250 ml of pure methanol and the collected eluate was concentrated under vacuum at 40 ° C and diluted with water and spray dried under the following conditions:

Spray dryer: cocurrent flow

Inlet temperature: 145 ° C

Outlet temperature 60 ° C

Nebulizer RPM 14000

The final weight is 4.5 gm.

Example 3

The powdered cinnamon powder having an average size of 1000 gm in the 16 mesh range was soaked in 2500 ml of butyl acetate and poured into an extractor having a 200 mesh mesh bottom sieve. The bottom extract was recirculated via the packaged material over a period of 10 hours to achieve an effective extraction. The eluent was discarded and the material was removed from the extractor and dried in a forced air oven at 30 °C. After the solvent is removed by evaporation, the material is reloaded into the extractor. The encapsulating material was extracted with acidified deionized water and the extract was recirculated through the bed at 30 ° C for about 12 hours to achieve an effective extraction.

The extract was filtered through a two-stage chromatography column to obtain a composition of 89% of a flavanoid type A proanthocyanidin pentamer (molecular weight: 1440), first passing the extract through the first column to extract the composition. The relatively low polarity molecules and the second stage chromatographic separation are used for relatively high polarity molecules in the composition. The resins used were equivalents of XAD-1180 and XAD-7HP resins, respectively. The column was thoroughly washed with deionized water to make it free of adhesive material and the eluent was neutral. The column was further eluted with 200 ml of pure ethanol and the collected eluate was concentrated under vacuum at 40 ° C and diluted with water and spray dried under the following conditions:

Spray dryer: cocurrent flow

Inlet temperature: 145 ° C

Outlet temperature 60 ° C

Nebulizer RPM 14000

The final weight is 4.8 gm.

Example 4

The powdered cinnamon powder having an average size of 1000 gm in the 16 mesh range was soaked in 2500 ml of butyl acetate and poured into an extractor having a 200 mesh mesh bottom sieve. The bottom extract was recirculated via the packaged material over a period of 10 hours to achieve an effective extraction. The eluent was discarded and the material was removed from the extractor and dried in a forced air oven at 30 °C. After the solvent is removed by evaporation, the material is reloaded into the extractor. The packaged material was extracted with 5 liters of acidified deionized water and the extract was recirculated through the bed at 30 ° C for about 12 hours to achieve an effective extraction.

The extract was filtered through a two-stage chromatography column to obtain a composition of 99% flavanoid type A proanthocyanidin pentamer (molecular weight: 1440), first passing the extract through the first column to extract the composition. The relatively low polarity molecules and the second stage chromatographic separation are used for relatively high polarity molecules in the composition. The resins used were equivalents of XAD-1180 and XAD-7HP resins, respectively. The column was thoroughly washed with deionized water to make it free of adhesive material and the eluent was neutral. The column was washed with pure isopropanol and the collected eluate was concentrated under vacuum at 40 ° C and diluted with water and spray dried under the following conditions:

Spray dryer: cocurrent flow

Inlet temperature: 145 ° C

Outlet temperature 60 ° C

Nebulizer RPM 14000

The final weight is 5 gm.

Example 5

The powdered cinnamon (casnamon cassia) powder having an average size of 1000 gm in the 16 mesh range was soaked in 3000 ml of ethyl acetate and poured into an extractor having a 200 mesh mesh bottom sieve. The bottom extract was recirculated via the packaged material over a period of about 8 hours to achieve an effective extraction. The eluent was discarded and the material was removed from the extractor and dried in a forced air oven at 30 °C. After the solvent is removed by drying, the material is reloaded into the extractor. The packaged material was extracted with 5000 ml of acidified deionized water at pH 4.0 and the extract was recirculated through the bed at 35 ° C for about 8 hours to achieve an effective extraction.

The extract was filtered through a two-stage chromatography column to obtain a composition having 55% of a flavanoid type A proanthocyanidin pentamer (molecular weight: 1440), and the extract was passed through the first column to extract the composition. The relatively low polarity molecules and the second stage chromatographic separation are used for relatively high polarity molecules in the composition. The resins used were equivalents of XAD-1180 and XAD-7HP resins, respectively. The column was thoroughly washed with deionized water to make it free of adhesive material and the eluent was neutral. The column was eluted with 175 ml of pure isopropanol and the collected eluate was concentrated under vacuum at 40 ° C and diluted with water and spray dried under the following conditions:

Spray dryer: cocurrent flow

Inlet temperature: 140 ° C

Outlet temperature 60 ° C

Nebulizer RPM 14000

The final weight is 2.5 gm.

Example 6: Extraction from dry litchi of lychee:

The 1000 gm powdered dried litchi peel was soaked in a 5000 ml volume of acidified water for 12 hours and clarified by filtration. The clarified filtrate was passed through a column containing an adsorption resin equivalent to XAD-1140 and XAD-7HP to separate polar and relatively non-polar compounds. The non-polar first column was eluted with ethanol and the eluate was concentrated to obtain a free-flowing powder with a yield of 500 mgm. A second column containing all polar substances alone was eluted with ethanol and concentrated to obtain 1 gm of powder. When analyzed by HPLC, this fraction showed 85% of a flavanoid type A proanthocyanidin pentapolymer (molecular weight: 1440).

Example 7: Extraction of peanut shells from red skin with peanut seeds

1000 gm of dried peanut shell with red skin on the seeds was soaked in a 5000 ml volume of acidified water at pH 3.8 and held for 48 hours and then filtered to a clear liquid. The clarified filtrate was passed through a column containing an adsorption resin equivalent to XAD-1140 and XAD-7HP to separate polar and relatively non-polar compounds. The non-polar first column was eluted with ethanol and the eluate was concentrated to obtain a free-flowing powder with a yield of 20 gm. A second column containing all polar materials alone was eluted with ethanol and concentrated to obtain 500 mg of powder. When analyzed by HPLC, this fraction showed 82% of the flavanoid type A proanthocyanidin pentamer (molecular weight: 1440), as shown in FIG.

Example 8: Purification to obtain flavanoid pentamers

The powder separated by the procedure detailed in Examples 1-6 was dissolved in 200 volumes of water and clarified by filtration. The clear filtrate was treated with activated charcoal at 60 ° C to decolorize the solution and clarified by filtration through a filter paper to remove all insoluble particles. The thus obtained filtered solution was extracted twice with ethyl acetate to remove all soluble solvent and concentrated to obtain a powder. The powder was subjected to column chromatography on a reverse phase C-18 tannin using a 0.1% aqueous formic acid solution and 0.1% methanolic acid in a flash chromatograph using the following parameters.

Device: Combiflash Companion with Variable UV Detector

Column: Redisep 12 gm (reverse phase cerium oxide)

Detection wavelength: 254 nm and 280 nm

Flow rate: 18 ml/min

Peak tube volume: 18 ml

Peak width: 1 min

Threshold: 0.20 AU

Solvent A: 0.1% formic acid aqueous solution

Solvent B: acetonitrile with 0.1% formic acid

Discard the fractions numbered 1 to 19. Fractions numbered 20 to 22 were pooled and concentrated to obtain 256 gm of light brown powder. TLC Screening Solvent System 0.1 M Sodium Acetate: Acetonitrile = 7:3 ratio The resulting powder exhibited a UV absorption spot at 0.75 Rt after spraying with an orange spotted anise/sulfuric acid reagent, which was considered to be a property of proanthocyanidins.

The EI-MS (M-H) (shown in Figure 2) ion peak at m/z 1439.9 corresponds to a plurality of intervals (288 heavy peaks) of the catechin structure corresponding to a total of five units. The molecular weight of the isolated compound was 1440.9. Confirmed by the coupling pattern, the configuration of the catechin unit is consistent with the configuration of epicatechin. The two signals between the four signals of δ 4.84 to 4.91 of the single peak of 2,3 cis stereochemistry (widened by the high molecular weight oligomer) in the flavan-3-ol unit. Between the peak broadening of δ 2.6 to 2.9 m observed due to high molecular weight oligomeric properties, the F-ring signal at the 4-position of the terminal unit -CH2-methylene proton was observed. For both systems with rings B and E, the aromatic region signal is between δ 6.6 and 7.6. The 13C carbon signal visible at δ 100.9 confirmed the C 2 carbon, and the 13 C carbon signal visible at δ 27.9 confirmed the C 4 carbon, and the top ring C confirmed the formation of a double bond with the intermediate system ring, as shown in FIG. 3 .

Obtaining pentameric proanthocyanidin flavonoids ranging from about 55% w/w to about 99% w/w, each from about 0.5% w/w to about 35% w/w, was obtained according to Examples 1 through 8 above. Compositions of terpolymers and tetramers in the concentration range, and optionally medically acceptable excipients. Further, the in vitro and in vivo activities described in the following Examples 9 to 14 were tested using the composition.

Example 9: Effect of test composition on body fluid antibody price in individuals treated with cyclophosphamide (immune insufficiency)

Swiss albino mice of any sex were divided into 5 groups based on the treatments they received, with 6 animals per group.

On day 0, sheep red blood cells (SRBC) containing 1 × 10 ⁸ cells in physiological saline were sensitized to all 5 groups. Groups 2-5 were treated with standard drugs and test composition combinations for 8 days (Day 0 to Day 7). On day 7, blood samples were collected from each of the 5 groups by posterior orbital puncture for determination of antibody titer. Then on the 7th day, after the blood was drawn, the mice were challenged by injecting 0.1 ml of SRBC into the footpad. On day 14, blood samples were collected from each mouse by posterior orbital puncture for determination of secondary antibody titers.

The results of this test are as follows:

Humoral immunity acts as the first line of defense and protects the host from infection. Increased humoral immunity makes protection against infectious pathogens stronger. The individual's response to the antigen varies and depends on a number of factors, including genetic composition and individual medical history. Cyclophosphamide is a drug that suppresses the immune system. Using this drug, the response of all animals evaluated in this study to the antigen can be identified.

As shown in the above table, the test composition reflects an increase in the cost of primary antibodies and secondary antibodies in individuals with immunocompromised individuals. The rate of primary antibody and secondary antibody (humoral immunity) is increased by a multiple. This immune response occurs in response to the presence of antigen, SRBC and considers changes in the immune response resulting from pretreatment with cyclophosphamide.

Example 10: Effect of test composition on phagocytic activity of peritoneal macrophages and blood multinucleated cells

The immune response to the antigen, although effective, requires further evaluation to determine the effectiveness of this challenge. The effectiveness of the pathogen is assessed by the immune response. The ability to eliminate pathogens is determined by assessing the phagocytic capacity of the reaction.

Swiss albinism mice of any gender were divided into 3 groups of 6 animals each. A single dose of control and test composition was administered to each of the 3 groups.

Each group was treated for a period of 20 days and on day 21, it received 5 ml of cold phosphate buffered saline (PBS) intraperitoneally. Next, the peritoneal fluid was collected and the number of macrophages was counted using a WBC square in a hemocytometer. Cell counts per cubic millimeter are estimated. The remaining mice remained in continuous treatment until day 28 and on day 29, blood was drawn from the posterior orbital plexus. Two drops of blood drawn from each mouse were placed on a slide and allowed to coagulate, and then placed in a moisturizing chamber, held at 37 ° C for 25 minutes. These adherent cells were incubated with a suspension of Candida albicans spores and observed by staining. Next, the number of cells ingesting the Candida suspension was counted.

The phagocytic index and phagocytosis % were calculated using the following formula:

Phagocytosis% = number of PMN cells ingesting Candida per 100 observed cells

Take the above steps and list the results:

The above results show an increase in peritoneal macrophage count and phagocytic activity. The increased number of macrophages directly indicates an enhanced immune response to the antigen. In addition, the increased phagocytic activity of these macrophages indicates their effectiveness against the antigen.

Example 11: Effect of test composition on host resistance to E. coli-induced abdominal sepsis

Swiss albinism mice of any gender were divided into 3 groups and treated with controls and test compositions. A single dose was administered to the mice for 28 days. On day 29, mice were intraperitoneally injected with a suspension of E. coli containing 2.5 x 10 ⁹ cells in PBS. After the injection, the mortality of the mice was observed over 24 hours. The observed mortality is caused by E. coli infection (also known as sepsis). The mortality of surviving animals was further observed over 7 days.

The results of this experiment are listed in the table below:

As can be seen from the above results, the mortality rate in the case of the control and the lower dose test composition was 100%. Mortality was reduced by 63% at the 50 mg/kg dose of the test composition. Only 3 of the 8 mice died compared to the death of all 8 mice in the other two groups. No further mortality was observed in this third group.

This display test composition provides a preventive effect to the host. Pretreatment with test compositions reduced mortality in mice exposed to pathogens. This confirms the ability of the test composition to prevent infection by pathogens, including bacteria and viruses.

This infection prevention is caused by an improvement in the immune response caused by the composition in the presence of a pathogen. This reaction allows the host to control the number of pathogens and thus prevent infection.

Example 12: Inhibition of influenza A (H1N1 and H3N2) viruses

This example demonstrates the efficacy of the test composition to inhibit H1N1 and H3N2 viruses and thus demonstrates its effectiveness as a method of treating, preventing, and managing this viral infection.

Martin-Dalby canine kidney (MDCK) cells were seeded into 6-well plates (3 x 10 ⁵ cells per well) and infected with H1N1 and H3N2 viruses the next day. After 3 days, cells were infected with serially diluted H1N1 and H3N2 viruses for 1 hour, followed by addition of 3 mL of overlay medium to each well. After 40 hours, cells were fixed with 10% formalin for 1 hour and stained with 1% crystal violet for 15 minutes. The viral power price was determined based on the plaque count.

The sensitivity of the virus to the compound was determined by a plaque reduction assay. The procedure was similar to the plaque assay except that the indicated amount of compound was added to the overlay medium. The percent inhibition is calculated as [100-(V _D /V _C )] x 100%, where V _D and V _C refer to the viral power price in the presence and absence of the compound, respectively. Dose of test plaques produced by self - Regression Analysis calculated response curves of reducing the number of the desired compound plaques minimum concentration of 50% (EC _50).

As is apparent from the above table, infected cells treated with the test composition exhibited a significant decrease in plaque formation. In addition, the test composition demonstrated a highly potent anti-H1N1 Tamiflu resistant strain, thus demonstrating its effectiveness as a possible treatment for influenza (H1N1) infection. In addition, the second table shows that the test compound is also effective against the H3N2 strain of influenza A virus.

Example 13: Effect of test composition on PBMC-stimulated HIV-1 (X4 tropic virus and R5 tropic virus)

HIV-1 (HXB2-X4 tropic) virus was obtained 48 hours after transfection of the HXB2 molecule to the 293T cells. The viral power price is detected by real-time PCR. Next, peripheral blood mononuclear cells (PBMC) stimulated by phytohemagglutinin (PHA) in 24 wells were infected with the virus. After 16-18 hours, PBMC were washed with phosphate buffered saline (PBS) and fresh Roswell Park Memorial Institute (RPMI) medium with 2% fetal bovine serum (FBS) was added. Cells and virus soup were collected on days 3, 5, and 7 (dpi) after infection.

Test composition and 3 standard drugs (AZT, AMD3100 and Tak- in PBMC cells stimulated by HXB2 HIV-1 virus (X4 virus and R5 virus) phytohemagglutinin (PHA-2 μg/mL) 779) Tested. The infection multiplication rate (MOI) was found to be 0.26. On day 3, day 5 and day 7 post infection, viral load detection was performed by RT-PCR as described above.

AMD3100 presented only for the inhibition of CXCR4 nutritional virus activity (average EC ₅₀ = 2.05 nM), and Tak-779 presented only for the inhibition of CCR5 nutritional virus activity (average EC ₅₀ = 0.56 nM). AZT exhibits inhibitory activity against CCR5 nutritional virus and CXCR4 nutritional virus. Test composition shows 22.5 μg / mL in the case of X4 virus (15.625 nM) EC ₅₀ values of ₅₀ and shows the value of 15.5 μg / mL (10.77 nM) EC in the case where R5 virus. These values are comparable and in some cases more effective than the standard drugs used.

Examples 9-11 present the immunoreactive properties of the test compositions, while Examples 12-13 show the antiviral effects of the test compositions against HIV and influenza virus (H1N1). In summary, Examples 9-13 demonstrate that the test composition is used to prevent infection by a two-pronged mechanism: the enhanced immune response can serve together as a protective or prophylactic option to prevent infection. Second, in the case of infection, the antiviral properties of the test composition reduce the viral load (both HIV and influenza), allowing an enhanced immune response to eliminate the infection and prevent further damage. It is important to note that this immune response is only triggered in the presence of antigen. This is evidenced by the fact that no animal exhibits signs of inflammation and/or other symptoms associated with an over-activated immune system.

This property of the test composition makes it highly suitable for long-term use as a prophylactic to prevent infection.

The compositions of the present invention comprise not only pentameric proanthocyanidin flavonoids ranging from about 55% w/w to about 99% w/w in a wide concentration range, from about 0.5% w/w to about 35% w. The immune response is improved in the presence of a composition of terpolymers and tetramers in the /w concentration range. The efficacy of the composition to elicit an immune response comprises from about 80% w/w to about 99% w/w of a pentameric proanthocyanidin flavonoid, each in a range of from about 0.5% w/w to about 20%, in a particular concentration range. The composition of the terpolymer and the tetramer in the w/w concentration range is better/excellent.

Example 14: Conceptual validation study for demonstrating the efficacy and safety of novel compositions in HIV and AIDS patients:

Scheduled, double-blind, randomized, placebo-controlled studies were performed in 40 patients who did not undergo antiretroviral, asymptomatic HIV-1 infection. 40 not subjected to the anti-retroviral, asymptomatic HIV-1 infected patients (whose CD4 count between 250-500 / mm ³⁾ in research and testing the composition. The test composition was tested at 300 mg/day for 12 weeks and administered in a capsule dosage form. The test composition exhibited a 11.29% reduction in viral load compared to a placebo showing a 67.28% increase in viral load. There was a decrease in CD4 counts in both groups, but the percent reduction in CD4 counts in the test composition group was found to be one and a half of placebo (7.74% in the case of test composition compared to 13.88% in the case of placebo). Therefore, the test composition was found to be safe and effective relative to most vital organ functions and biochemical parameters. Example 14 details the ability of the test composition to inhibit viral load to confirm the antiviral effect of the test composition. Again, the improvement in WBC (CD4) counts is an important indicator of improved immune response compared to placebo.

Figure 1 shows the molecular structure of the flavonoid pentamer.

Figure 2 shows the EI-MS of pentameric pentamers of flavonoids.

Figure 3 shows the 13C NMR of the flavonoid pentamer.

Figure 4 shows flash chromatography of the composition to identify flavonoid pentamers.

Figure 5 shows the HPLC of the flavonoid pentamer.

Claims (26)

A composition comprising a pentameric proanthocyanidin flavonoid in a concentration range of 55% w/w to 99% w/w, a terpolymer each in a concentration range of 0.5% w/w to 35% w/w And tetramers, and optionally medically acceptable excipients. The composition of claim 1, wherein the composition is obtained from a plant source selected from the group consisting of Cinnamomum , Litchi , and Arachis . The composition of claim 1, wherein the concentration of the pentameric proanthocyanidin flavonoid is preferably in the range of 80% w/w to 98% w/w, and the trimer and the tetramer are each preferably. It is in the range of 0.5% w/w to 20% w/w. The composition of claim 1, wherein the pentameric proanthocyanidin flavonoid has a molecular weight of about 1440; and the pentamer is a type A proanthocyanidin pentamer. The composition of claim 1, wherein the excipient is selected from the group consisting of a gum, a granulating agent, a binder, a lubricant, a disintegrating agent, a sweetener, a coloring agent, a flavoring agent, a coating agent, A group of plasticizers, preservatives, suspending agents, emulsifiers, antistatic agents, and spheronizing agents. The composition of claim 1, wherein the composition is formulated from the group consisting of a tablet, a sugar-coated tablet, a buccal tablet, an aqueous or oily suspension, a dispersible powder or granule, a hard gel capsule or a soft gel. Various dosage forms of the group of emulsions, syrups and elixirs in gel capsules. For example, the composition of claim 1 is used to improve the immune response of a disease in an individual in need thereof, and the disease is selected from the group consisting of epidemic sex. Groups of predators, HIV infections and AIDS. The composition of claim 7, wherein the composition is in a medically effective amount within a weight range of 1 mg/kg of the individual to 100 mg/kg of the individual; and the composition is for treatment, Preventing and managing an infection caused by a pathogen in the individual, the pathogen comprising an influenza A virus selected from the group consisting of influenza virus H1N1 and H3N2; and selected from HIV X4 and R5 tropic virus ) The group of HIV viruses. The composition of claim 7, wherein the individual is an animal or a human. The composition of claim 1 is for the treatment, prevention and management of a viral infection in an individual in need thereof, the viral infection being selected from the group consisting of influenza A virus, HIV X4 tropism and R5 orientation. A group of sexual viruses. The composition of claim 10, wherein the composition is in a medically effective amount of from 1 mg/kg body weight to 100 mg/kg body weight of the individual; and the individual is an animal or a human. A method of preparing a composition comprising pentameric proanthocyanidin flavonoids in a concentration range of 55% w/w to 99% w/w, each in a concentration range of 0.5% w/w to 35% w/w a trimer and a tetramer, the method comprising the steps of: a) pulverizing a plant source to obtain a pulverized plant body and extracting the pulverized plant body using an organic solvent to remove toxic substances; b) drying the plant Body to remove the organic solvent; c) re-extracting the dried plant body with acidified deionized water to obtain an extract; and d) purifying the extract via a two-stage chromatography column, followed by concentration, purification, standardization and drying to obtain the composition . The method of claim 12, wherein the plant source and the powdered plant system are selected from the group consisting of plants of the genus Cinnamon, Litchi, and Peanut. The method of claim 12, wherein the organic solvent is selected from the group consisting of ethyl acetate, butyl acetate, amyl acetate, 2-ethylhexyl acetate, and any combination thereof. The method of claim 12, wherein the extracting is carried out for a period of from 8 hours to 12 hours. The method of claim 12, wherein the extracting is carried out for 10 hours. The method of claim 12, wherein the toxic substance comprises coumarin and an aldehyde. The method of claim 12, wherein the re-extraction is carried out at a pH in the range of 3.8 to 5.8; and the re-extraction is carried out in the range of from 30 ° C to 90 ° C for a period of from 8 hours to 12 hours. The method of claim 12, wherein the re-extraction is carried out at a pH of 4.0; and the re-extraction is carried out at a temperature ranging between 31 ° C and 40 ° C for 10 hours. The method of claim 12, wherein the chromatography column is selected from the group consisting of XAD-1180, XAD-7HP, and XAD-1140 resins. The method of claim 12, wherein the composition further comprises a gum, a granulating agent, a binder, a lubricant, a disintegrating agent, a sweetener, a coloring agent, a flavoring agent, a coating agent, and a coating agent. Excipients of the group of plasticizers, preservatives, suspending agents, emulsifiers, antistatic agents, and spheronizing agents. a pentameric proanthocyanidin flavonoid comprising a concentration ranging from 55% w/w to 99% w/w, and a trimer and tetramer in each concentration ranging from 0.5% w/w to 35% w/w The use of a composition of matter for the manufacture of a medicament for improving the immune response of a disease in an individual in need thereof, the disease being selected from the group consisting of influenza, HIV infection and AIDS. The use of claim 22, wherein the pharmaceutical product provides a medically effective amount of the composition in a weight range from 1 mg/kg of the individual to 100 mg/kg of the individual; and the composition is used for Treating, preventing, and managing an infection caused by a pathogen in the individual, the pathogen comprising an influenza A virus selected from the group consisting of influenza viruses H1N1 and H3N2; and selected from HIV X4 and R5 tropic viruses A group of HIV viruses. The use of claim 22, wherein the individual is an animal or a human. a pentameric proanthocyanidin flavonoid comprising a concentration ranging from 55% w/w to 99% w/w, and a trimer and tetramer in each concentration ranging from 0.5% w/w to 35% w/w The use of a composition of matter for the manufacture of an antiviral agent for the treatment, prevention and management of a viral infection in an individual in need thereof, the viral infection being selected from the group consisting of influenza A virus, X4 tropism of HIV and R5 A group of sexual viruses. The use of claim 25, wherein the antiviral preparation provides a medically effective amount of the composition in a weight range of from 1 mg/kg of the individual to 100 mg/kg of the individual; and the individual is an animal Or human.