JP2006213609A - Activating agent of immunocompetent cell, method for preventing feline immunodeficiency virus infection by using the same, method for eliminating feline immunodeficiency virus, method for preventing cancer growth and method for eliminating cancer - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a new activating agent of immunocompetent cells, capable of activating animal immunocompetent cells. <P>SOLUTION: This activating agent of the immunocompetent cells contains propolis having a function of activating the immunocompetent cells both quantitatively and qualitatively as an active ingredient. <P>COPYRIGHT: (C)2006,JPO&NCIPI

Description

  The present invention relates to an activator for immunocompetent cells. More specifically, the present invention relates to a method for preventing feline immunodeficiency virus infection, a method for eliminating feline immunodeficiency virus, a method for preventing cancer occurrence, and a method for destroying cancer.

  Propolis is a dark brown and sticky resinous material made by mixing bees' own saliva and secretions containing enzymes into a complex of gum, sap, perfume, etc. collected by bees from trees. It is. Originally, honeybees are applied to the nest walls to strengthen the nest walls and prevent bacteria from growing in the nests. Propolis is said to have antibacterial action, antiviral action, antipyretic action, etc. for a long time, and has been used for a long time as a folk remedy.

  The main components of this propolis are flavonoids, phenols such as aromatic acids, cinnamic acid, aromatic oils, etc., and other composites that contain various components. Different. For example, Brazilian propolis is mainly composed of artepillin C, which is a cinnamic acid derivative, and Chinese propolis is mainly composed of flavonoids.

Recent pharmacological studies have shown that the effects of propolis are not only antibacterial and antiviral, but also antitumor (anticancer), immunoregulatory, and blood pressure lowering. It has been proposed to be used for food (for example, Patent Document 1 and Patent Document 2). Propolis has been reported to promote the expression of IFN-γ in mouse spleen lymphocytes and the expression of other cytokines such as IL-2 and IL-4. It is considered.
JP 2003-128561 JP JP 2001-333731 A

  However, the only animal whose immunomodulatory action of propolis has been confirmed so far is only a mouse. For example, in the inventions of Patent Documents 1 and 2, the target animal is only a mouse, and the action of other animals is as follows. The actual situation is that no concrete data on the effect is shown. That is, it can be said that the detailed mechanism of the immunomodulatory action of propolis in animals other than mice has not been elucidated.

  As a result, in the field of veterinary medicine, it is also used in research on the prevention and treatment of diseases of many animals other than mice (domestic animals such as pigs and horses, pets such as cats and dogs), especially immunodeficiencies and cancer. Because of its influence, there is a demand for the development of new drugs and methods for the treatment of immunodeficiency animals, especially cat immunodeficiency virus (FIV) infections, and cancer. Nevertheless, no successful development has been reported yet.

  Therefore, the present invention was made in view of the circumstances as described above, solves the conventional problems, can quantitatively and qualitatively activate animal immunocompetent cells, The object is to provide a new activator of immunocompetent cells. In addition, a novel feline immunodeficiency virus infection prevention method, feline immunodeficiency virus infection method, and cancer prevention method and cancer, which can activate the immune function of animals to prevent or treat FIV infection and cancer. It is an object to provide a method for destroying cells.

  The invention of this application is as a means for solving the above-mentioned problems. First, the present invention contains a propolis having a function of quantitatively and qualitatively activating immunocompetent cells as an active ingredient. The second feature is that the main component of propolis is a cinnamic acid derivative, the third feature is that the cinnamic acid derivative is artepilin C, and the fourth is Furthermore, it is characterized by containing a pharmacological component, and fifth, the immunocompetent cell is a cat.

  Furthermore, the present invention sixthly relates to the feline immunodeficiency virus (FIV) infection by activating and improving feline immunity by administering any of the first to fifth inventions to the feline. And, seventhly, the immunity of the cat is activated by administering any of the first to fifth inventions to a cat suffering from a feline immunodeficiency virus (FIV) infection. It is characterized in that it is improved by removing the FIV.

  Furthermore, according to the present invention, eighthly, by administering any of the first to fourth inventions to an animal, the immunity of the animal is activated and improved, and the occurrence of cancer is prevented. Ninth, the animal is a cat.

  Tenth, the invention is characterized in that the first to fourth inventions are administered to an animal in which cancer has occurred, thereby activating and improving the immunity of the animal and destroying cancer cells. And eleventh, the animal is a cat.

  According to the first to third inventions, animal immunocompetent cells can be activated quantitatively and qualitatively.

  According to the fourth invention, in addition to the effects of the first to third inventions, animal immunocompetent cells can be activated more efficiently and quantitatively.

  According to the fifth invention, in addition to the effects of the first to fourth inventions, cat immunocompetent cells can be activated quantitatively and qualitatively.

  According to the sixth invention, feline immunodeficiency virus (FIV) infection can be prevented by the effects of the first to fourth inventions.

  According to the seventh invention, FIV can be driven out by the effects of the first to fourth inventions.

  According to the eighth invention, the occurrence of cancer can be prevented by the effects of the first to third inventions.

  According to the ninth aspect, the effect of the eighth aspect can be applied to a cat.

  According to the tenth invention, cancer cells can be driven out by the effects of the first to third inventions.

  According to the eleventh aspect, the effect of the tenth aspect can be applied to a cat.

  The present invention has the features as described above, and the embodiments thereof will be described in detail below.

  The present invention has been made based on the finding that propolis activates immunocompetent cells quantitatively and qualitatively as a result of intensive studies by the present inventors. To explain further, propolis promotes lymphocyte blastogenesis and IFN-γ expression. That is, the immunocompetent cell activator of the present invention comprises propolis having such characteristics as an active ingredient. In the activator of the present invention, the propolis content is not particularly limited, but is, for example, 0.001 to 20.0% by weight, preferably 0.01 to 10.0% by weight of the activator.

  Propolis usually contains various components such as various flavonoids, various diterpenes, various minerals, various vitamins, various cinnamic acid derivatives, etc., but the propolis in the present invention is particularly p-coumaric acid or artepilin. It is preferable that a cinnamic acid derivative such as C is contained as a main component, and it is more preferable that the cinnamic acid derivative is artepilin C because it activates immunocompetent cells more efficiently.

  Artepilin C is usually represented by the following formula (1)

It is represented by

  The activator of the present invention may further contain a pharmacological component to activate the immunocompetent cells more efficiently and to facilitate administration (or ingestion). Moreover, it can be set as functional food-drinks (immunostimulation supplementary food) which are easy to administer or ingest by mixing and processing with food-drinks components (or food-drinks itself). The food and drink component can be mixed and processed without particular limitation as long as it can maintain the effects of the present invention. Examples thereof include seasonings such as soy sauce and miso, and beverages such as fruit juice juice. Furthermore, it can be mixed with animal feeds such as livestock and pets, and the mixing amount (blending amount) at that time is, for example, 1 to 1000 mg / kg as propolis, and the effects of the present invention are sufficiently obtained. Can be demonstrated.

  These pharmacological components are various carriers that are generally used for drug production. The carrier is appropriately selected from a solid carrier or a liquid carrier depending on the type of drug and the administration form such as injection or oral. It means what can be done. For example, oral liquid drugs such as suspensions and syrups are water, sugars such as sucrose, glycols such as polyethylene glycol, oils such as sesame oil and soybean oil, etc., antiseptics, peppermint, etc. These can be produced using various flavors, gelatin, starch and the like. Powders, pills, capsules and tablets include excipients such as lactose, glucose and sucrose, disintegrants such as starch, lubricants such as magnesium stearate, various binders and surfactants, plasticizers It can be formulated using a stabilizer or the like.

  With the activator of the present invention having such characteristics, it is possible to efficiently and quantitatively activate the immunocompetent cells of animals including humans. As a result, various cells such as bacteria and viruses can be activated. It exerts preventive and destroying effects against infectious diseases and various cancers. This immunocompetent cell refers to a cell involved in a series of immunity reactions, and includes, for example, macrophages, T cells, B cells, NK cells, Tc cells, and the like. Quantitative and qualitative activation of immunocompetent cells means increasing the amount of immunocompetent cells (quantitative activation) and increasing the function of immunocompetent cells (qualitative activation). ) Means.

  As an action mechanism in which the activator of the present invention activates immunocompetent cells, the macrophage is IL-1α, IL-1β, IL-2, after the propolis in the activator of the present invention is phagocytosed by the macrophage. It promotes the expression of various cytokines such as IL-4 and IFN-γ, thereby activating immune functions such as T cells, B cells, NK cells, and Tc cells, which are immunocompetent cells.

  The animal to be used for the activator of the present invention (that is, the immunocompetent cell to be activated) is not particularly limited, regardless of whether it is livestock, pets, wild animals or the like. For example, birds such as chickens, reptiles such as lizards, amphibians such as frogs, fish such as zebrafish, and mammals such as humans, cats, monkeys, pigs, cows, sheep, mice (mouse), horses, etc. Particularly preferred is a mammal. Furthermore, since the activator of the present invention activates feline immunocompetent cells and activates immunocompetence, the animal to be used is preferably a cat among mammals.

  The activator of the present invention can also be used in humans, can activate human immunocompetent cells and improve immunity, and has a preventive effect on various infectious diseases and cancers. Destroying effect.

  The propolis, which is an active ingredient of the activator of the present invention, is a preparation produced by honey bees such as honey bees and bee honey bees, but may be secretions of other kinds of bees. Further, the production area is not limited. Examples of production areas include Japan, China, Southeast Asia, Europe, the Americas, Africa, etc. For example, the base plants from China and Europe are poplar and pine, and the base plants from Brazil are eucalyptus and acrelen trees. It is. Among them, Brazil is preferably produced from Brazil because the propolis in the activator of the present invention is based on an arlenic tree containing abundant Artepilin C as a base plant. The propolis may be a sample collected from a bee captured in nature or a sample collected from an artificially raised bee. Furthermore, in order to remove the impurities and increase the purity of the sample, a processed sample subjected to a treatment such as separation or extraction may be used.

  By using such an activator, infectious diseases caused by bacteria or viruses can be prevented or eliminated (treated). In particular, the activator of the present invention can activate feline immunocompetent cells and efficiently improve immunity by administering to cats, thereby preventing feline immunodeficiency virus (FIV) infection. can do. In addition, administration to a cat suffering from FIV infection can also efficiently activate and improve the immunity of the cat, and as a result, drive out FIV.

  In addition, since the activator of the present invention can be administered to an animal to activate the immunocompetent cells of the animal and improve the immunity, the activated immunocompetent cells can be efficiently transformed into early cancerous cells. Can be eliminated immunologically to prevent the development of cancer. In addition, this principle of improving immunity is sufficiently exerted even in animals in which cancer has occurred. That is, the activator of the present invention is administered to an animal suffering from cancer, and the immunocompetent cells of the animal are activated to improve immunity. As a result, activated immunocompetent cells can efficiently destroy cancer cells. At this time, as described above, the animal species to which the activator of the present invention is used is not particularly limited. However, the activator of the present invention activates feline immunocompetent cells in particular to improve immunity. When activated for the purpose of preventing or destroying cancer, the animal to be used is preferably a cat. The target cancers include lung cancer, liver cancer, stomach cancer, colon cancer, skin cancer, and the like, as well as those in which a plurality of cancers occur in addition to these single cancers.

  The dose (use amount) varies depending on the animal species, age, administration route, number of administrations, etc. of the administration target, and can be appropriately changed, and is not particularly limited. For example, an effective amount administered as a combination of an effective amount of propolis and a suitable diluent and pharmacological component (a pharmacologically usable carrier) is 100-5000 mg / kg body weight per day, 6 The dose is divided into 24 hours.

  In consideration of facilitating absorption into the body, the propolis in the activator of the present invention is preferably extracted by known hot water extraction or alcohol extraction, but in addition, extraction by steam distillation or pressurized hot water (It is also referred to as “subcritical water”, generally hot water in a liquid state at 100 ° C. or higher, pressurized to a saturation vapor pressure or higher) and may be extracted by extraction separation or the like.

  The outline of the extraction can be explained by, for example, water (including hot water) or organic solvent (for example, petroleum ether, cyclohexane, toluene, carbon tetrachloride, dichloromethane, chloroform, diethyl ether, diisopropyl ether, acetate ether, butanol, n -Propanol, ethanol, methanol, polyethylene glycol, propylene glycol, pyridine, etc.), or a mixed solvent of two or more of these, and usually obtained by extraction at 3 to 70 ° C. These extracts can be used as they are, or appropriately diluted or concentrated, or dried.

  Regardless of animal species, immunocompetent cells of many animals can be activated and immunity can be improved by the activator of the present invention having the above characteristics and the method of using the activator. . As a result, it is effective in preventing and destroying (treating) various infectious diseases caused by bacteria, viruses, etc., particularly FIV infectious diseases, and can prevent the occurrence of cancer and destroy (treat) cancer cells.

  EXAMPLES Hereinafter, although an Example demonstrates this invention further more concretely, this invention is not limited to these Examples.

Example 1: Preparation of propolis The propolis in the activator of the present invention is a product of Brazil extracted by alcohol extraction, which is a known extraction method.
Example 2: Examination of action effect on cats As test animals, 1 to 10 year old healthy cats (4 heads: 2 males, 2 females) were used. (1) in vitro
Concanavalin A (ConA) was added as a propolis and stimulant to mononuclear cells isolated from feline peripheral blood and cultured, and lymphocyte blastogenicity was examined by the MTT method, which is a cell proliferation test.

  The results were as shown in FIG. 1 and FIG.

  FIG. 1 is a diagram showing the results of examination of lymphocyte blastogenesis of propolis in the activator of the present invention, (A) shows the relationship between the concentration of propolis and the lymphocyte blastogenesis ability, and (B) It shows the degree of lymphocyte rejuvenation by propolis.

  As shown in FIG. 1, when the propolis concentration was 300 μg / ml, the absorbance increased, indicating that the lymphocytes became immature. Further, in the propolis alone addition group (propolis in the figure) and the ConA + propolis addition group (propolis + ConA in the figure), a significant increase in absorbance was observed compared to the control group (PBS in the figure). In particular, a significant increase was observed in the group supplemented with propolis alone.

  Table 1 shows the absorbance values corresponding to FIG.

  FIG. 2 shows the results of comparative examination of stimulation index values (SI values) in the control group (ConA in the figure), the propolis alone addition group (propolis in the figure), and the ConA + propolis addition group (propolis + ConA in the figure). FIG.

  SI value is the following formula (2)

Calculated and compared.

  As shown in FIG. 2, in all groups, an increasing tendency was observed as compared with the symmetric group, and in particular, when propolis was added alone, a marked increase was observed.

  Table 2 shows a list of calculated SI values.

(2) in vivo
Propolis 0.05 mg / kg was orally administered to 3 healthy cats for 7 days, and the lymphocyte rejuvenation ability and the expression of cytokine IFN-γ were examined before and after administration. The presence or absence of the stimulant Concanavalin A (ConA) was also examined.

  The lymphocyte blastogenicity was examined by the MTT method, and the expression of IFN-γ was examined by the RT-PCR method.

  The results are as shown in FIG. 3 and FIG.

  FIG. 3 is a diagram showing the examination results of lymphocyte blastogenicity before and after administration of propolis, (A) shows the results of examination by absorbance before and after administration, and (B) shows the results of examination by SI value. ing. FIG. 4 is a diagram showing the results of studying the expression of IFN-γ before and after administration of propolis. (A) shows the case without ConA mixing, and (B) shows the case with ConA mixing.

  As shown in FIG. 3, both the absorbance and the SI value showed an increasing tendency after administration as compared to before administration of propolis. Table 3 shows the absorbance values in FIG. 3 (A), and Table 4 shows the SI values calculated in FIG. 3 (B).

  Moreover, as shown in FIG. 4, it was confirmed that the expression of IFN-γ was remarkably promoted only by adding propolis regardless of the presence or absence of addition of ConA. In particular, when ConA was added, a significant increase in IFN-γ expression was observed after administration compared to before administration of propolis.

  Table 5 shows the measured values of IFN-γ in FIG.

Example 3: Examination of effects on dogs The activator (active ingredient: propolis) of the present invention was added to 0.05 mg / kg of dogs (dog breed: golden retriever, age: 7 years, sex: female). Was orally administered for 1 week, and the lymphocyte rejuvenation ability before and after administration was examined. The examination method was performed by measuring the absorbance by the MTT method.

  The result was as shown in FIG.

  FIG. 5 is a diagram showing the results of examination of canine lymphocyte blastogenesis in the presence or absence of ConA mixing before and after administration of propolis in the activator of the present invention. As shown in FIG. 5, regardless of the presence or absence of ConA mixing, administration of propolis confirmed an increase in absorbance, that is, an increase in lymphocyte blastogenicity.

  Table 6 shows the absorbance values in FIG.

  From these results, propolis, which is an active ingredient of the activator of the present invention, improves immunity by promoting lymphocyte blastogenesis and IFN-γ expression regardless of animal species. I was able to confirm.

It is the figure which showed the examination result of the feline lymphocyte rejuvenation of the propolis in the activator of this invention, (A) is the relationship between a propolis addition density | concentration and the lymphocyte rejuvenation ability, (B) is the lymph by propolis. It shows the degree of ball juvenile ability. Comparison results of stimulation index values (SI values) in the control group (PBS in the figure), the propolis alone addition group (propolis in the figure), and the ConA + propolis addition group (propolis + ConA in the figure) in cats FIG. It is the figure which showed the examination result of the feline lymphocyte rejuvenation ability before and after propolis administration in the activator of this invention, (A) is the result examined by the absorbance before and after administration, (B) is examined by SI value. Shows the results. It is the figure which showed the examination result of the expression of feline IFN-gamma before and after the administration of propolis in the activator of the present invention, (A) without ConA mixing, (B) with ConA mixing. Show. It is the figure which showed the examination result of canine lymphocyte rejuvenation in the case of the presence or absence of ConA mixing before and after administration of propolis in the activator of the present invention.

Claims (11)

  An activator for immunocompetent cells, comprising a propolis having a function of activating the immunocompetent cells quantitatively and qualitatively as an active ingredient.   The activator according to claim 1, wherein the main component of propolis is a cinnamic acid derivative.   The activator according to claim 1 or 2, wherein the cinnamic acid derivative is artepilin C.   The activator according to any one of claims 1 to 3, further comprising a pharmacological component.   The activator according to any one of claims 1 to 4, wherein the immunocompetent cell is a cat.   A feline immunodeficiency virus (FIV) infection is prevented by administering the activator according to any one of claims 1 to 5 to a cat to activate and improve feline immunity. How to prevent FIV infection.   The activator according to any one of claims 1 to 5 is administered to a cat suffering from a feline immunodeficiency virus (FIV) infection, thereby activating and improving the feline's immunity and destroying FIV. A method for destroying FIV, characterized in that   A method for preventing the occurrence of cancer, comprising administering the activator according to any one of claims 1 to 4 to an animal to activate and improve the immunity of the animal to prevent the occurrence of cancer. .   The prevention method according to claim 8, wherein the animal is a cat.   A cancer cell characterized in that the activator according to any one of claims 1 to 4 is administered to an animal in which cancer has occurred, thereby activating and improving the immune ability of the animal and destroying the cancer cell. Destroying method.   The destroying method according to claim 10, wherein the animal is a cat.

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