WO2006008761A2

WO2006008761A2 - Anti retroviral herbal formulation

Info

Publication number: WO2006008761A2
Application number: PCT/IN2005/000246
Authority: WO
Inventors: Raja A.K.
Original assignee: A K Raja
Priority date: 2004-07-23
Filing date: 2005-07-21
Publication date: 2006-01-26
Also published as: WO2006008761A3

Abstract

Anti retroviral herbal formulation comprising of synergistic combination of medicinal herbs characterized in that the extraction process involves the steps of obtaining fresh mature roots of Gycyrrihza glabra, bulbs of Allium sativum, seed kernels of Azadirachta indica, and pulverizing the same; extracting the fine powder obtained from pulverizing, with cold water and collecting the filtrate and air dried; preparing acetic acid solution from the extract of Piper betel, Pichcorhiza kurroa, Annona Squamosa, Witania Somnifera and Tinospora Cordifolia; soaking of the above obtained air dried filtrate with extracted acetic acid solution at normal room temperature and filtered.

Description

Antiretro Viral Herbal Formulation

Field of the Invention: This invention pertains generally to herbal extracts, methods and treatments, and particularly relates to a herbal extract composition and method having therapeutic immunity boosting and antiviral effects for boosting or stimulating the natural immune response in humans.

Background of the invention: The spread of HIV in India has been diverse, with much of India having a low rate of infection and the epidemic being most extreme in the southern half of the country and in the far north-east. The highest HIV prevalence rates are found in Maharashtra in the west; Andhra Pradesh and Karnataka in the south; and Manipur and Nagaland in the north-east. As of May 2005, 92% of all nationally reported AIDS cases have been found in 10 of the 38 States Union Territories. The greatest numbers were in Maharashtra and Gujarat in the west; Tamil Nadu and Andhra Pradesh in the south; and Manipur in the north-east. India has the second highest number of people living with HIV/AIDS in the world after South Africa. India accounts for almost 10 per cent of the 40 million people living with HIV/AIDS globally and over 60% of the 7.4 million people living with HIV/AIDS (PLWHA) in the Asia and Pacific region. India is facing one the biggest public health challenges in its history. Several modern medications have been developed from ancient healing traditions associated with specific plants. The medicinal properties of many plants have been identified with specific chemical compounds, which have been isolated, purified and, in many cases, synthetically reproduced. Many well known drugs were originally derived from plants. Salicylic acid, the precursor for aspirin, was originally isolated from white willow bark and the meadowsweet plant. Quinine, which is used to treat malaria, was derived from Cinchona bark. Vincristine, which is used in cancer treatment, comes from periwinkle. The cancer drug Taxol was originally isolated from the bark and needles of the Pacific Yew Tree. Perhaps most famous are morphine and codeine, which are derived from the opium poppy. Morphine is still the standard against which new synthetic pain relief drugs are measured.

A number of compositions particularly nucleoside analog reverse Transcription inhibitors (NTRIs) and protease inhibitors are available, however the costly compositions prepared therefrom are out of reach of the common patients in the developing countries. These compounds also have severe toxic effects. Till date no effective treatment for these diseases has been found and attempts are being made by the mankind to reduce the hardships of the patients suffering from these disease. Various chemical and other allopathic medicines are available for treatment of these diseases. The human immune response is extremely complex and requires constant interaction by the cells involved in the immune system. Intracellular communication is facilitated by secretion of chemical messenger proteins known as cytokines, which act by enhancing cell growth, promoting cell activation, directing cellular traffic, stimulating macrophage function, destroying antigens, and other functions. Cytokines generally comprise lymphokines, which are secreted by T-lymphocytes, and monokines secreted by monocytes.

Persons who suffer from immuno-suppressive diseases or conditions such as human immunodeficiency virus (HIV) or experience decreased levels of lymphocytes and correspondingly decreased levels of cytokines. For example, where individuals have been infected by HIV, lymphocytes having a cell surface antigen known as CD4 are present in noticeably low numbers. While in healthy individuals these CD4 lymphocyte cells are present in concentrations of about 800 cells per milliliter of serum, HIV-infected individuals exhibit as few as 200 CD4 cells per milliliter of serum when opportunistic infections develop. Similarly in case of CD8 lymphocyte the concentration is found to be very less in case of HIV affected person.

Modern physicians, rely on treatments using synthetic or chemically manufactured drugs rather than plant derived or herbal extracts for treatment. Pharmacologists tend to identify, isolate, extract and synthesize the active compounds from plants for use in treatment. This approach, however, has drawbacks i.e. in addition to individual physiologically active compounds present in a plant, there are also minerals, vitamins, glycosides and other substances which are important in supporting the medicinal properties of a plant to provide a synergistic effect. Generally these additional substances is absent when purified. The toxicity of the purified physiologically active compounds is also generally high.

Various herbs are believed to have a beneficial effect on the human immune system. For example, Echinacea Purpurea and Echinacea Angustifolia are believed to stimulate T-cell activity. Mowery, D. B.; The Scientific Validation of Herbal Medicine, Keats Publishing, Inc., New Canaan Press, 1986. P. 118-119. Astragulas Membranaceous is believed to be able to stimulate production of interferon and Immunoglobulins A and G (IgA and IgG) in mice. Kaiser, J. D.; Immune Power. A Comprehensive Treatment Programfor HIV. St. Martin's Press, New York, 1993, p. 59-60.

The efficacy of various herbal remedies, extracts and treatments is well known, and therapeutic herb products are increasingly recognized as desirable alternatives to synthetic drugs. U. S. Patent No. 5,178,865 discloses an herbal extract mix which inhibits infection of human immunodeficiency virus or HIV in vitro. U. S. Patent No. 5,500,340 and U. S. Patent No. 5,294,443 disclose the use of herbal extracts for immunosuppression and treatment of autoimmune disorders. The international patent WO 02/32444 provides herbal compositions for treating patients with liver disease and HIV comprising various parts of different plants such as Herba Hedyotidis diffusae, Rhizoma Bistortae, Rhizoma Polygoni Cuspidati, and Fructus Schisandrae and so on and a method for treatment using the compositions mentioned therein.

However, a plant-derived or herbal extract which effectively provides a stimulative or boosting effect to the human immune system has heretofore been unknown.

Accordingly, there is a need for a synergistic herbal extract composition and method composition and method which provides a health supplement and for treatment generally persons having diseases or conditions which suppress the immune response.

Despite all of the available for the treatment of HIV, there is still no cure for the deadly disease. HIV viruses continue to mutate and become resistant to existing drugs such as the reverse transcriptase and protease inhibitors.

The present invention satisfies these needs, as well as others, and generally overcomes the deficiencies found in the state of art. Objectives: The main objective of this invention is to provide a formulation that effectively inactivates human retroviruses. Another objective of the invention is to provide a formulation that acts as a neutralizing antibody. Another objective of the invention is to boost immune system and block the action of reverse transcriptase. Yet another objective of the invention is to increase the count of CD 4 cells and CD 8 cells. Another object of the invention is to provide an herbal extract composition and method which provides a nutritious dietary supplement for humans and animals. Another object of the invention is to provide an herbal extract composition and method which stimulates cell mediated immunity in humans and animals.

Further objects and advantages of the invention will be brought out in the following portions of the specification, wherein the detailed description is for the purpose of fully disclosing the preferred embodiment of the invention without placing limitations thereon.

Summary of the invention: The present invention is a synergistic herbal extract composition and method of making and using the same for stimulating or boosting the immune response in humans and animals. The present invention involves a novel formulation of herbal extracts obtained from Glycyrrhiza Glabra, Picorhiza Kurroa, Azadirachta Indica, Annona Squamosa, Specific type of AMn Sativum, Tinospora Cordifolia, Withania Sominifera and Piper Betle. The specific parts of these herbal plants are collected, cleaned and shade dried. The dried herbs are powdered and extracted using predetermined process. The extract is then filtered. The active principles of the individual antiviral herbal medicines or medicinal plants or extracts thereof have been isolated through specific isolation techniques and have been characterized through the use of accepted chemical techniques. The active molecule in the filtrates is then processed in systematic process to obtain the desired novel formulation. The formulation obtained in this invention effectively inactivates human retroviruses and boosts immune system. The composition of the formulation is non-toxic and can be taken orally as a drug or vaccine, by both children and adults, to reinforce the immune system against virus, without interfering with most other medications. The novel formulation inactivates further growth and replication of the retrovirus thus increasing the count of CD 4 cells and CD 8 cells.

Further objects and advantages of the invention will be brought out in the following portions of the specification, wherein the detailed description is for the purpose of fully disclosing the preferred embodiment of the invention without placing limitations thereon. The names, classifications, and geographic origins of the plants or herbs used with the invention are provided below in

TABLE 1

Detailed description of the invention: The present invention pertains generally to the use of herbal extracts of eight major herbs and various combinations thereof for boosting or stimulating the immune response in humans and animals, for treatment of persons and animals suffering from immunosuppressive and auto-immune diseases or conditions, and for inhibiting HIV infection in humans.

Extract Preparation The following describes the preparation and combination of extracts of Glycyrrhiza Glabra, Allium Sativum, Azadirachta Indica, Piper Betle, Picohiza Kurroa, Annona squamoa, Withiania Somnifera and Tinsopora Cordifolia. The processes described below can be scaled up to produce larger quantities of extracts. The details provided for preparation of the following abstracts reflect the presently preferred method for extract preparations and should not be considered as limiting. The quantities and times described below can be varied substantially to provide suitable extracts of Glycyrrhiza Glabra, Allium Sativum, Azadirachta Indica, Piper Betle, Picohiza Kurroa, Annona squamoa, Withiania Somnifera and Tinsopora Cordifolia. in accordance with the invention.

The process extraction involves the following steps of: obtaining fresh mature roots of Gycyrrihza glabra and cleaning the obtained roots with one liter of distilled water. The cleaned roots are air dried at room temperature for about 4 - 5 hrs; obtaining Bulb of Allium sativum and the outer coat is removed. The cleaned bulbs are air dried at room temperature for about 4 - 6 hrs.; obtaining seed kernels of Azadirachta indica which are 3 to 8 months old. The seed kernels are air dried at room temperature for about 2 - 3 days and the outer coat is removed; obtaining the air dried roots of Glycyrrhiza glabra, Bulb of Allium sativum , seed kernels of Azadirachta indica and pulverizing the same for about 15 - 30 mins at a medium rpm; extracting the fine powder obtained from pulverizing, with cold water and collecting the filtrate in a glass beaker and air dried; preparing acetic acid solution from the extract of Piper betel, Pichcorhiza kurroa, Annona Squamosa, Withania Somnifera and Tinospora Cordifolia; soaking of the above obtained air dried filtrate with extracted acetic acid solution for about 12 - 20 hours at normal room temperature and filtered.

The extract comprises of extracts from herbs Gycyrrihza glabra, Allium sativum, Azadirachta indica, Piper betel, Pichcorhiza kurroa, Annona Squamosa, Withania Somnifera and Tinospora Cordifolia with active ingredients and mixed in proportion without using any external solvents. The said extracts are present to each other in relative ratio of: about 5% by weight to about 20% by weight of Glycyrrhiza glabra, about 5% by weight to about 20% by weight of Allium sativum, about 5% by weight to about 20% by weight of Azadirachta indica, about 3% by weight to about 10% by weight of Piper betle, about 3% by weight to about 10% by weight Picorhiza Kurroa, about 3% by weight to about 10% by weight of Withania Somnifera, about 3% by weight to about 10% by weight of Tinospora Cordifolia.

The extracts are in specific ratio of Glycyrrhiza glabra 10% - 18%, Allium sativum 10% - 18%, Azadirachta indica 10% - 18%, Piper betle 3% - 8%, Picorhiza Kurroa 3% -8%, Withania Somnifera 3% - 8%, Tinospora Cordifolia.3% -8% and active ingredients extracted. The air dried roots of Glycyrrhiza glabra, Bulb of Allium sativum , seed kernels of Azadirachta indica and pulverizing the same for about 15 - 25 mins at a medium rpm to obtain powder of size 10 mesh sieve to 30-mesh sieve. The soaking of the obtained air dried filtrate with extracted acetic acid for about 12 - 18 hours at normal room temperature and filtered. The anti retroviral herbal formulation as claimed the herbal composition is in non-encapsulated, powder form, capsule form. The formulation is a base of berries or fruit or a base of vegetable soup or a milk drink or a nutritive supplement. A method of treating HIV infected individual consists of administration of the anti retroviral synergistic herbal formulation to boost and stimulates the immune response in humans.

The following experimental results on acute oral toxicity indicate that the herbal extract composition of the invention is effective at inhibiting HIV infection in vitro. These experiments and the resulting data demonstrate that the herbal extract composition of the invention is non-toxic and is a useful source or candidate for potential immune disorder therapies.

Immunomodulatory test

The following experimental results on immunomodulatory activity indicate that the herbal extract composition of the invention is effective at inhibiting HIV infection in vitro. These experiments and the resulting data demonstrate that the herbal extract composition of the invention is a useful source or candidate for potential immune disorder therapies.

The experiment was conducted to test the immunomodulatory activity of LBT/gatp-1/04. Wistar albino rats. Either sex (3 male + 3 female), weighing (150-18Og) were used for the studies. The animals were housed in polypropylene cages in a temperature controlled room at 25±20C with 40 - 50% relative humidity. The animals were kept at 12:12 hrs dark : light schedule. They were pelleted laboratory animal feed and tap water ad libitum. Wistar rats of either sex ( 120 ± 15 g) were divided into four groups of six animals each and each group received the following regimen of treatment. Group I - Control rats ( Propylene Glycol) Group Il - Adjuvant arthritic rats, Group III - LBT/gatp-1/04 alone.

The immunomodulatory activity were verified based on Candida phagocytosis ( Wilkinson, 1977) test and Nitro blue tetrazolium reduction test (NBT). The Candida test results proved that the number of positive cells per 100 neutrophils is high. The count of neutrophils gives the phagocytic index. The NBT test performed by isolating granulocytes from blood by incubating 0.5ml of heparinized blood over a clean acetone dried glass slide at 370C for 30 minutes in a moist chamber. Due to the inherent adhering property the neutrophils adhere to the glass slide. The blood was then gently washed away with sterile cold normal saline delivered by a capillary pipette. The edge of the slide was gently tapped to remove excess saline, but not allowed to dry. Immediately the incubating test medium was poured on the slide, which consists of 0.2 ml of NBT, 0.2 ml f 0.34% sucrose in saline and 0.2 ml of pooled serum. A cover slip was gently placed over this to provide a thin layer and good contact and incubated at 370C for 30 minutes. After 30 minutes cover slip was gently removed, excess fluid drained and the slide dried. The slide was fixed with methanol for 3 minutes, washed in distilled water, dried completely and was stained with safranin for 7 minutes, washed in distilled water, dried and examined under oil immersion 100 cells were observed and the positive cells for the formazon were counted. Thus the NBT test proves that LBT/gatp-1/04 activities the immune regulation and is capable of increasing the phagocytosing of lymphocytes, which is beneficial in the treatment.

The above experiments and resulting data also demonstrate that the herbal extract composition of the invention is a useful potential source or candidate for anti-HIV and other antiviral therapies and treatments for humans and animals suffering from HIV, other viral infections, or complications or disorders associated with such viral infections.

The anti-HIV activity of the herbal extract composition of the invention may contain a single pharmacological active ingredient, component or agent acting alone, or a combination of such ingredients, components or agents, and/or biological metabolites or derivatives thereof. In vivo therapies and treatments using the herbal extract composition of the invention for anti- HIV and antiviral treatments will likely be based on orally ingested or dosages of the herbal extract composition in liquid or solid form. Rectal, parenteral, intravenous, aerosol inhalation or subcutaceous routes for in vivo administering of the herbal extract composition of the invention are also possible. As for treatment of immunosuppresive treatments, the herbal extract composition may also be administered in vivo for antiviral treatments in admixture or combination with appropriate excipients, carriers, antiviral agents, immune modulators, chemotherapeutic agents, antibodies, antiviral agents or combinations thereof. Pharmacological preparations of the invention may be dosage unit forms such as tablets, capsules, suppositories, lotions, ampoules or metered liquid or aerosol dosages.

The herbal extract composition of the invention provides a healthy and nutritious supplement to the human diet including various cytokine or cytokine-like proteins, and immunoglobulin or immunoglobulin-like proteins, which are believed to provide a stimulating or boosting effect to the immune system. As nutrition supplement the herbal extract composition of the invention is ingested orally, or may be applied topically.

The dosage of the compositions of the invention obtained from the invention can range from 0. 5 to 125 g per day for the mammal in need of therapy. One skilled in the art will appreciate that depending upon the weight of the individual and the progression of the viral infection, that higher doses of the compositions may be required. As the compositions according to the invention have demonstrated virtually no side effects, high doses may be initiated with reduction of dosage upon manifestation (i. e., reduction of viral load) of therapeutic effect. One skilled in the art can tailor each dosage rate for a given individual without undue experimentation. More specifically, the dosages for a given composition can range from 0. 4 to 100 g per day, more preferably 1.0 to 5Og per day. Preferably, the compositions are administered at least three (3) times per day, however, bolus administration will be effective.

Accordingly, it will be seen that this invention provides an herbal extract composition that stimulates the lymphocyte to inhibit HIV infection, and boosts or stimulates the immune response in humans. Although the description above contains much specificity, these should not be construed as limiting the scope of the invention but as merely providing an illustration of the presently preferred embodiment of the invention. Thus the scope of this invention should be determined by the appended claims and their legal equivalents.

While the invention has been described in connection with specific embodiments thereof, it will be understood that it is capable of further modifications and this application is intended to cover any variations, uses, or adaptations of the invention following, in general, the principles of the invention and including such departures from the present disclosure come within known or customary practice within the art to which the invention pertains and may be applied to the essential features hereinbefore set forth.

References

1. Atal CK, Sharma ML, Kaul, Khajuria A. Immunomodulating agents of plant origin. l:preliminary screening. J Ethnopharmacol 1986; 18:133-441

2. Mowery, D. B.; The Scientific Validation of Herbal Medicine, Keats Publishing, Inc., New Canaan Press, 1986. P. 118-119.

3. Kaiser, J. D.; Immune Power. A Comprehensive Treatment Program for HIV. St. Martin's Press, New York, 1993, p. 59-60.

Claims

I claim:

1. An Anti retroviral herbal formulation comprising of synergistic combination of medicinal herbs characterized in that the extraction process involves the steps of obtaining fresh mature roots of Gycyrrihza glabra, bulbs of Allium sativum, seed kernels of Azadirachta indica, and pulverizing the same; extracting the fine powder obtained from pulverizing, with cold water and collecting the filtrate and air dried; preparing acetic acid solution from the extract of Piper betel, Pichcorhiza kurroa, Annona Squamosa, Withania Somnifera and Tinospora Cordifolia; soaking of the above obtained air dried filtrate with extracted acetic acid solution at normal room temperature and filtered.

2. The anti retroviral herbal formulation as claimed in claim 1 wherein the extract comprises of extracts from herbs Gycyrrihza glabra, Allium sativum, Azadirachta indica, Piper betel, Pichcorhiza kurroa, Annona

Squamosa, Withania Somnifera and Tinospora Cordifolia with active ingredients and mixed in proportion without using any external solvents.

3. The anti retroviral herbal formulation as claimed in claim 1-2, wherein the said extracts are present to each other in relative ratio of: about 5% by weight to about 20% by weight of Glycyrrhiza glabra, about 5% by weight to about 20% by weight of Allium sativum, about 5% by weight to about 20% by weight of Azadirachta indica, about 3% by weight to about 10% by weight of Piper betle, about 3% by weight to about 10% by weight Picorhiza Kurroa, about 3% by weight to about 10% by weight of Withania Somnifera, about 3% by weight to about 10% by weight of Tinospora cordifolia.

4. The anti retroviral herbal formulation as claimed in claim 1-3, wherein extracts are in specific ratio of Glycyrrhiza glabra 10% - 18%, Allium sativum 10% - 18%, Azadirachta indica 10% - 18%, Piper betle 3% - 8%, Picorhiza Kurroa 3% -8%, Withania Somnifera 3% - 8%, Tinospora cordifolia.3% -8% and active ingredients extracted.

5. The anti retroviral herbal formulation as claimed in claim 1 , wherein the air dried roots of Glycyrrhiza glabra, Bulb of Allium sativum , milky white seed kernels of Azadirachta indica of 3 to 8 months old, plain absolute yellow fruit, and pulverizing the same for about 15 - 25 mins at a medium rpm to obtain powder of size 10 mesh sieve to 30-mesh sieve,

6. The anti retroviral herbal formulation as claimed in claim 1 , wherein soaking of the obtained air dried filtrate with extracted acetic acid for about 12 - 18 hours at normal room temperature and filtered.

7. The anti retroviral herbal formulation as claimed in claim 1 , wherein the herbal composition contains a newly identified protein that may inhibit HIV virus in vitro

8. The anti retroviral herbal formulation as claimed in claim 1 , wherein the herbal composition contains active principles that can inhibit the RNA virus.

9. The anti retroviral herbal formulation as claimed in claim 1 and claim 8, wherein the herbal composition contains active principles such as Chlorogenic Acid or Oic Acid or Glyxrrhizin that can inhibit the HIV virus.

10. The anti retroviral herbal formulation as claimed in claim 1 , wherein the herbal composition contains pytochemicals in the formulation that are capable of stimulating and strengthening the body's immune system, scavenging free radicals and detoxifying carcinogens.

11. The anti retroviral herbal formulation as claimed in claim 1 - 5, wherein the herbal composition is in non- encapsulated, powder form.

12. The anti retroviral herbal formulation as claimed in claim 1 - 5, wherein the herbal composition is in capsule form

13. The anti retroviral herbal formulation as claimed in claim 1 - 5, wherein said formulation is a base of berries or fruit or a base of vegetable soup or a milk drink or a nutritive supplement.

14. A method of treating HIV infected individual consists of administration of the anti retroviral synergistic herbal formulation of claim 1 - 5 to boost and stimulates the immune response in humans.