JP4464964B2 - Treatment for polycystic ovary syndrome (PCOS) - Google Patents

Description

  The present invention relates to an effective and safe therapeutic agent for polycystic ovary syndrome (PCOS), which causes infertility, menstrual abnormalities, acne, hirsutism, obesity in women, and the like.

  Recent studies have reported that 6% of women of reproductive age have polycystic ovary syndrome (PCOS). And the number of patients with potential polycystic ovary syndrome (PCOS) is believed to be much higher. Polycystic ovary syndrome (PCOS) refers to a condition in which the follicle does not steadily mature large until ovulation, and the ovary is filled with a large number of small immature eggs, causing ovulation disorders.

  Among the gonadotropic hormones (gonadotropins) secreted from the pituitary gland and required for ovulation, FSH (Follicle-stimulating hormone) and LH (luteinizing hormone) are the causes of this. It is thought that this is because the balance with is disturbed, but there are still many unexplained parts.

  Polycystic ovary syndrome (PCOS) causes symptoms such as irregular menstruation, amenorrhea, menorrhagia, increased male hormone (Testosterone), obesity, and causes infertility. The following pharmacotherapy and surgical treatment are known as conventional treatment methods for polycystic ovary syndrome (PCOS).

(I) Ovulation inducer (clomiphene, clomid) therapy Clomifen works on the hypothalamus to cause ovulation, but ovulation may not occur.

(II) Combination therapy with clomiphene and other drugs When there is no effect when only clomiphene is administered, it is a treatment method in which clomiphene is combined with a steroid hormone or a drug such as HMG (human menopausal gonadotropins).

(III) Gonadotropin therapy (HMG or FSH administration method)
Gonadotropin (gonadotropin) is injected into a patient and stimulates the ovary to induce ovulation. This treatment method is relatively effective and has a good pregnancy rate, but has a problem in that it tends to cause side effects such as multiple pregnancy and ovarian hyperstimulation syndrome (OHSS).

(IV) Method of administering Shakuyakukanzoto This is a method of administering Shakuyakukanzoto, which is a kind of Chinese herbal medicine, and has few side effects, but does not have a satisfactory effect.

(V) Laparoscopic treatment method This is a treatment method that uses a laparoscope to irradiate the surface of the ovary with a laser beam to open a small hole. It is not a radical treatment method, and there is a problem that invasiveness to the living body cannot be avoided.

(VI) Oral contraceptive regimen For unmarried and young women with polycystic ovary syndrome (PCOS) who do not wish to have a baby (pregnancy), use menstrual agents such as oral contraceptives to cycle menses The treatment that makes you wake up.

  For women with infertile polycystic ovary syndrome (PCOS) who want to have a baby, the treatment mainly aimed at ovulation is performed as described above. Among them, the treatment methods using ovulation inducers such as (I) to (III) are considered trumps for infertility treatment, but may cause serious side effects such as multiple pregnancy and ovarian hyperstimulation syndrome (OHSS). It has sex and poses difficult problems.

  There is another problem in the treatment of unmarried women and young women. Many of these patients do not want to see the obstetrics and gynecologists and may be left without appropriate treatment. If the disease is left as it is, there is a problem that a vicious circle occurs in which symptoms such as ovarian hypertrophy progress and the treatment becomes more difficult.

  In addition, even when consulting the obstetrics and gynecology department, doctors and patients should use strong drugs such as ovulation inducers such as (I) to (III) and gonadotropin for unmarried women and young women. Tend to avoid as much as possible. The administration of the above oral contraceptive (IV) is also resistant to the patient. Therefore, at present, there is a problem that only inadequate symptomatic treatment is performed under a diagnosis name such as “unscheduled menstruation”.

  Thus, the conventional treatment method for polycystic ovary syndrome (PCOS) has various problems as described above, and does not satisfy both the effectiveness and the absence of side effects. Furthermore, recent studies have reported that patients with polycystic ovary syndrome (PCOS) are seven times more likely to develop diabetes compared to unaffected individuals. As is well known, diabetes is a serious disease that causes myocardial infarction, stroke, renal failure, blindness due to retinopathy, and the like. Therefore, it is necessary to treat this at the stage of polycystic ovary syndrome (PCOS) before complications of diabetes.

  The present invention has been made in view of the above problems, has almost no side effects, is effective and improves anovulation, and can be used safely not only for women who want to have a baby, but also for unmarried and young women An object is to provide a possible therapeutic agent for polycystic ovary syndrome (PCOS).

  In order to solve the above-mentioned problems, the present inventors have conducted research on safe and effective therapeutic agents for polycystic ovary syndrome (PCOS). As a result, mushroom extracts have been developed into polycystic ovary syndrome (PCOS). The present invention was completed by finding effective activity.

That is, the present invention is a therapeutic agent for polycystic ovary syndrome (PCOS) comprising a maitake extract as an active ingredient.

In addition, the maitake extract (1) is obtained by treating maitake raw materials ( maitake fruit bodies and / or mycelium collectively as “ maitake raw materials”) with ethanol having a concentration of 90% or more to remove ethanol-soluble components. (2) Thereafter, hot water extraction is performed, ethanol is added to the obtained hot water extract until the final volume concentration is 50 to 75%, and the insoluble components produced are removed to obtain a supernatant. It is preferable to be produced by a process including a process and (3) a process of collecting a fraction having a molecular weight of 14,000 or more from the obtained supernatant.

  The therapeutic agent for polycystic ovary syndrome (PCOS) of the present invention is safe, has few side effects, and has an excellent effect of improving anovulation, by using an extract of mushrooms as an active ingredient. It can be used safely not only for women who want children, but also for unmarried and young women.

FIG. 1 is a chart showing the results of high performance liquid chromatography (HPLC) analysis of the maitake extract obtained in Production Example 1. (Analysis example)
FIG. 2 is a chart showing the results of high performance liquid chromatography (HPLC) analysis of shiitake extract obtained in Production Example 3. (Production Example 3)
[FIG. 3] FIG. 3 is a chart showing the results of high performance liquid chromatography (HPLC) analysis of the Agaricus extract obtained in Production Example 4. (Production Example 4)
[FIG. 4] FIG. 4 is a chart showing the results of high performance liquid chromatography (HPLC) analysis of the litchi extract obtained in Production Example 5. (Production Example 5)
FIG. 5 is a chart showing the results of high performance liquid chromatography (HPLC) analysis of the oyster mushroom extract obtained in Production Example 6. (Production Example 6)

  The therapeutic agent for polycystic ovary syndrome (PCOS) of the present invention comprises an extract of mushrooms as an active ingredient. This mushroom extract can be used as a treatment method for polycystic ovary syndrome (PCOS). In addition, this mushroom extract can be used to produce a therapeutic agent for polycystic ovary syndrome (PCOS).

  The mushrooms are used for food and medicine, and have been prized for their health since ancient times. albicans), shiitake mushrooms (Lentinus edodes), Agaricus (Agaricus blazei Murill), mushroom (Agaricus bispirus), Coffs key Polyporaceae (Ganoderma applanatum), hemlock Polyporaceae (Fomitopsis pinicola), Coriolus versicolor (Coriolus versicolor), litchi (Ganoderma lucidum , Oyster mushrooms (Pleurotus ostreatus), king oyster mushroom (Pleurotus eryngii), Hericium erinaceus (Hericium erinaceus), Cordyceps sinensis (Cordyceps sinensis), Semitake (Cordyceps sobolifera), fungus (Auricularia auricula), from the white fungus (Tremella fuciformis), and Phellinus (Phellinus linteus) It is preferable that it is 1 or more types chosen from the group which consists of.

  Among these, at least one selected from the group consisting of maitake, shiitake, agaricus, litchi and oyster mushrooms is more preferable, and maitake is most preferable.

  These mushrooms contain glycoproteins, which are sugar-bound proteins (complex proteins), and the extracts from these mushrooms according to the present invention contain this glycoprotein as the main component. It is thought that.

  Glycoprotein, which is the main component of the mushroom extract according to the present invention, is brown, exhibits water solubility and heat stability, and has a mass ratio of simple protein portion to carbohydrate portion of 75:25 to 90: 10. It is preferable that the molecular weight distribution is 18,000 to 22,000 and positive for the burette reaction and the Fering reaction. Here, the “simple protein portion” refers to a portion consisting only of a polypeptide chain in glycoprotein which is a complex protein.

  The molecular weight distribution of glycoprotein is preferably 18,000 to 22,000, and the average molecular weight is more preferably 20,000. Moreover, heat stability means that biological activity is not lost even if it is left at a temperature of 80 to 130 ° C. for 1 to 5 hours.

  The mushroom extract according to the present invention includes (1) a step of treating the mushroom raw material (mushroom fruit body and / or mycelium) with ethanol having a concentration of 90% or more to remove ethanol-soluble components; (2) Thereafter, hot water extraction is performed, ethanol is added to the obtained hot water extract until the final volume concentration is 50 to 75%, the insoluble components produced are removed, and a supernatant is obtained. 3) It is preferable to be produced by a process including a process of collecting a fraction having a molecular weight of 14,000 or more from the obtained supernatant.

  The method for producing an extract of mushrooms according to the present invention includes the steps (1) to (3). First, as the first step, (1) mushroom raw materials used in the present invention are treated with ethanol having a concentration of 90% or more to remove ethanol-soluble components.

  As the mushroom raw material used in the present invention, a fresh product of the mushroom raw material or a dried product thereof can be used without particular limitation. Among these, it is preferable to use a dry powder from the viewpoint of efficient treatment. Further, the ethanol having a concentration of 90% or more may be water (ethanol water) containing 90% or more of ethanol. Among them, water that contains 95% or more of ethanol is preferable, and ethanol is most preferably 100% from the viewpoint of efficiently removing ethanol-soluble components dissolved from mushroom raw materials.

  The treatment in the step (1) means adding ethanol or ethanol water to a mushroom material and stirring. The content of the mushroom raw material at this time is preferably 10 to 25% by mass, and more preferably 10 to 12.5% by mass. Moreover, although processing temperature shall be 20-70 degreeC, room temperature may be sufficient. The treatment time depends on the state of the mushroom raw material used, the treatment temperature, and the like, but is preferably 1 to 10 hours, and more preferably 2 to 3 hours.

  By this treatment, components soluble in ethanol from the mushroom raw material are dissolved in ethanol or ethanol water. Since the dissolved ethanol-soluble component is unnecessary, it is removed by filtration or centrifugation. Among them, it is preferable to use centrifugation from the viewpoint of mass processing.

  Next, as the step (2), hot water extraction is performed from the ethanol extraction residue from which the ethanol-soluble components have been removed, and ethanol is added to the obtained hot water extract until the final volume concentration is 50 to 75%. The components are removed and a supernatant is obtained.

  The hot water extraction is to add water to the ethanol extraction residue obtained in the step (1) and heat it. The concentration of the ethanol extraction residue at this time is preferably 10 to 25% by mass, and more preferably 10 to 12.5% by mass. Moreover, it is preferable that heating temperature is 80-130 degreeC, and it is more preferable to carry out at 100-120 degreeC. Moreover, it is preferable that extraction time is 1 to 5 hours, and it is more preferable that it is 2-3 hours.

  Next, components insoluble in this solution are filtered off, and ethanol is added to the hot water extract, which is the filtrate after filtration, until the final volume concentration is 50 to 75%. It may be efficient to concentrate the hot water extract to 1/3 to 1/2 of its volume before adding ethanol.

  After adding ethanol, it is allowed to stand at a low temperature, preferably 0 ° C. to room temperature, more preferably 4 to 10 ° C., for 5 to 24 hours, preferably 8 to 12 hours. It is preferable to promote the generation of. Thereafter, the generated insoluble component is removed by centrifugation or filtration, and a supernatant containing a filtrate or the like is obtained. By removing insoluble components produced by the addition of ethanol, substances having immunosuppressive action and the like can be substantially removed from the final product.

  Next, as a step (3), a fraction having a molecular weight of 14,000 or more is collected from the obtained supernatant. The collection of fractions having a molecular weight of 14,000 or more is preferably performed using dialysis or ultrafiltration. The dialysis membrane used for dialysis or ultrafiltration is not particularly limited as long as it has a molecular weight cut-off of 14,000, and general membranes such as cellophane membranes and collodion membranes can be used.

  Thereafter, the obtained fraction having a molecular weight of 14,000 or more may be purified for the purpose of further increasing the purity. As such a purification method, for example, a method often used for purification of glycoprotein such as gel filtration chromatography can be used without particular limitation.

  The solvent used in the manufacture of the above mushroom extract is approved by the Japanese Ministry of Health, Labor and Welfare for use in the production of health food materials, etc., and this extract can also be used as a health food material. is there.

  The therapeutic agent for polycystic ovary syndrome (PCOS) of the present invention can contain a pharmaceutically acceptable carrier and / or diluent in addition to the above-mentioned extract of mushrooms as an active ingredient. As such a carrier, for example, cellulose, calcium monohydrogen phosphate, sucrose fatty acid ester, silicon dioxide, methylcellulose, lactose and the like can be used. Moreover, as a diluent, water, glycerol, etc. can be used, for example. In addition, general additives such as preservatives, stabilizers, excipients, binders, disintegrants, and sweeteners may be added.

  The therapeutic agent for polycystic ovary syndrome (PCOS) of the present invention can be administered orally, parenterally or transdermally. In general, the amount of the active ingredient to be administered is desirably determined appropriately according to the weight of the patient, the nature of the disease, the condition, the administration route, and the like. Among them, it is preferably 50 to 800 mg / person orally per day, more preferably 100 to 500 mg / person, and most preferably 200 to 350 mg / person. Divide and administer.

  The extract of mushrooms according to the present invention can be used on its own or with other pharmacologically active substances. Forms suitable for administration include, for example, tablets or coated tablets, capsules, suppositories, solutions, syrups, emulsions, or dispersible powders. The tablet or the like preferably contains 3 to 80% by mass, and more preferably 5 to 50% by mass, of the mushroom extract according to the present invention.

  Tablets include, for example, one or more active substances known excipients, eg, inactive ingredients such as calcium carbonate, calcium phosphate, lactose; disintegrants such as starch, alginic acid; binders such as starch, gelatin; stearic acid Lubricants such as magnesium and talc; and / or agents that give delayed release such as carboxymethyl cellulose, cellulose acetate phthalate, and polyvinyl acetate, and extracts of the mushrooms according to the present invention, or other pharmacologically active agents It can be produced by mixing the substance. A tablet may consist of several layers.

  The coated tablet can be produced by coating a core produced in the same manner as a tablet with a substance usually used for tablet coating, such as gum arabic, talc, titanium dioxide, methylcellulose, sucrose and the like. The core to obtain delayed release and to avoid incompatibility may consist of several layers. Similarly, the tablet coating may consist of several layers to give a delayed release, and the above excipients for tablets can be used.

  A syrup containing an extract of the mushrooms according to the present invention or a combination thereof with other pharmacologically active substances is a sweetener such as saccharin, cyclamate, glycerol, sucrose, and a flavor improving agent, For example, flavoring agents such as vanillin and orange extract may be contained. Further, it may contain a suspending aid or thickener such as sodium carboxymethylcellulose; a wetting agent such as a condensate of an aliphatic alcohol and ethylene oxide; or a preservative such as p-hydroxybenzoate.

  In addition, the capsule containing the extract of mushrooms according to the present invention or a combination thereof with other pharmacologically active substances is prepared by mixing these substances with an inert carrier such as lactose or sorbitol. The mixture can be produced by enclosing the mixture in a gelatin capsule or the like.

  Further, suppositories are, for example, extracts of the mushrooms according to the present invention, or combinations of this with other pharmacologically active substances, such as conventional carriers, specifically neutral fat, polyethylene glycol or derivatives thereof, etc. It can manufacture by mixing with.

  In addition, the mushroom extract according to the present invention or a combination of this with another pharmacologically active substance can be a food such as health food, functional food or general food. Since the glycoprotein in the present invention is an extract of mushrooms, it is excellent in safety, and the food can be ingested constantly. These foods may contain vitamins, minerals, herbs and other nutritional materials in addition to the above-mentioned additive components.

  The following examples are illustrative without limiting the scope of the invention.

[Production Example 1] Production of mushroom (maitake) extract To 1000 g of dried maitake fruit body powder, add 5 L of 95% ethanol water, treat at room temperature for 2 to 3 hours, and remove ethanol-soluble components by filtration. did.

  Subsequently, 5 L of deionized water was added to the obtained residue, and extraction was performed for 2 hours while stirring at 100 to 120 ° C. The hot water extract was concentrated to a volume of about 1/2, and ethanol was added until the final volume concentration was 50-75%. This was allowed to stand for 8 to 12 hours in a low temperature room at 4 to 10 ° C., and then solid substances such as precipitates and suspended matters as insoluble components were removed by centrifugation to obtain a supernatant.

  From this supernatant, fractions having a molecular weight of 14,000 or more were collected by dialysis to obtain a maitake extract.

  In order to analyze this maitake extract, it was purified by gel filtration chromatography to obtain about 21 g of brown material. The purified maitake extract was subjected to a burette reaction and a failing reaction. As a result, it was confirmed that the purified maitake extract contained a protein portion and a carbohydrate portion.

[Analysis Example] Analysis of purified maitake extract The simple protein portion of the purified maitake extract obtained as described above was quantified by the Bradford method. The analysis of the constituent amino acids of the simple protein portion was performed by an automatic amino acid analyzer (Hitachi L-8500A Amino Acid Analyzer). The saccharide portion was quantified by the phenol-sulfuric acid method, and the saccharide portion composition was determined by high performance liquid chromatography (HPLC). The molecular weight was measured by SDS-PAGE electrophoresis. ¹ H-NMR measurement was also performed.

  Table 1 shows the analysis results of the mass ratio between the simple protein portion and the carbohydrate portion in the purified maitake extract. The substance was found to be a glycoprotein.

  In addition, FIG. 1 shows a chart of analysis results of high-performance liquid chromatography (HPLC) of the maitake extract obtained in Production Example 1. In FIG. 1, the peak at 7.189 was identified as glycoprotein.

The glycoprotein which is a main component in the obtained maitake extract had the following properties.
Appearance: Brown, hygroscopic powder Solubility: Soluble in water and alkaline solution Stability: Stable to high temperature, stable to ethanol Chemical composition: Simple protein part: Carbohydrate part = 75:25 to 90:10 (Table 1 reference)
Amino acid composition of simple protein part: Asparagine, glutamine, serine, threonine, glycine, alanine, valine, cysteine, methionine, isoleucine, leucine, tyrosine, phenylalanine, lysine, histidine, arginine, proline Composition of carbohydrate part: galactose, mannose , Glucose, N-acetylglucosamine, fucose
¹ H-NMR spectrum: 3.0473, 3.8893, 6.8500, 7.3150 Average molecular weight: 20,000.

[Production Example 2] Tablet Production Method Using the maitake extract obtained in Production Example 1, tablets were produced with the following composition.
Tablet per tablet Main ingredient:
Maitake extract (powder) 36mg
250mg dry maitake powder
Auxiliary:
Microcrystalline cellulose 264mg
Phosphate-calcium hydrogen 20mg
Sucrose fatty acid ester 20mg
Atomized silicon dioxide 10mg
Coating material:
Methylcellulose 1-1.5% of tablet mass

  Here, the dried maitake powder is a powder obtained by drying and pulverizing maitake, and is not subjected to ethanol extraction. The above-mentioned maitake extract (powder) and dried maitake raw material powder are mixed with a finely pulverized auxiliary, granulated, dried, crushed, tableted, made into tablets of appropriate shape and size, and then coated The material was applied.

The analysis result of the obtained tablet and the result of the hygiene inspection are shown below. This tablet was confirmed to satisfy the standards of the Pharmaceutical Affairs Law.
Analysis items:
Disintegration time 45 minutes or less Mass difference 600mg ± 5%
Hygiene inspection:
Number of general viable bacteria 3000 or less Yeast and mold 300 or less Salmonella negative Escherichia coli group negative

[Production Examples 3 to 6] Manufacture and analysis of extracts from shiitake mushrooms, agaricus, litchi, oyster mushrooms, mushrooms instead of maitake, shiitake (manufacturing example 3), agaricus (manufacturing example 4), litchi (manufacturing example 5) A mushroom extract was produced in the same manner as in Production Example 1, except that oyster mushroom (Production Example 6) was used. These extracts were subjected to HPLC measurement in the same manner as in the above analysis example. FIG. 2 is a chart of HPLC analysis results of shiitake extract, FIG. 3 is a chart of HPLC analysis results of agaricus extract, FIG. 4 is a chart of HPLC analysis results of litchi extract, and FIG. The charts of the HPLC analysis results of the oyster mushroom extract are shown respectively. 2-5, it is in the substantially same position as 7.189 of the maitake extract of manufacture example 1, ie, 7.087 (FIG. 2), 7.082 (FIG. 3), 7.096 (FIG. 4), 7 respectively. A peak was observed at 0.086 (FIG. 5), and it was confirmed that the same components as the maitake extract of Production Example 1 were contained.

[Test Example 1] Administration to patients with polycystic ovary syndrome (PCOS) Outpatients diagnosed with polycystic ovary syndrome (PCOS), 18 anovulatory patients over 3 months randomly divided into 2 groups divided. In the test group, the tablet produced in Production Example 2 (the tablet of the present invention) was administered in 9 tablets a day in 3 divided doses, and in the control group, 7.5 g of Shakuyakukanzoto was administered 3 times a day. Divided into two doses. The administration was continued for 12 weeks (3 cycles), and the basal body temperature table was recorded for each group of patients to confirm the presence or absence of ovulation.

  Table 2 shows the result of comparison of the ovulation rate according to the number of ovulated patients. Table 3 shows the result of comparison based on the number of cycles ovulated.

  According to Table 2, when comparing the ovulation rate in the case where each of the tablets produced in Production Example 2 (the tablet of the present invention) and Shakuyakukanzoto is administered in terms of the number of patients, the tablet of the present invention: Shakuyakukanzoto = 75%: 10 %Met. Moreover, according to Table 3, when compared with the number of cycles of ovulation, the present invention tablet: Shakuyakukanzoto = 46%: 10%. According to Fisher's exact method, P <0.05 and a significant difference was observed. Statistically, the treatment agent for polycystic ovary syndrome (PCOS) of the present invention is significantly more significant than the treatment with Shakuyakukanzoto. It was confirmed to be excellent. Further, no side effects were observed in both groups, and it was confirmed that the therapeutic agent for polycystic ovary syndrome (PCOS) of the present invention is highly safe.

[Test Example 2] Safety test of mushroom extract A safety test was conducted using 10 healthy 4-week-old ICR mice, each male and female. After measuring the body weight of these mice 4 hours after fasting, the test group was prepared by dissolving the maitake extract obtained in Production Example 1 in distilled water for both males and females at a dose of 2,000 mg / kg body weight. Forced single administration using a sonde. In the control group, 0.7 mL of male water and 0.6 mL of female pure water were similarly administered. The observation period was 14 days, and all were necropsied after the observation period.

As a result, neither male nor female died during the observation period, and there was no abnormality in general condition and body weight. At necropsy, no abnormalities were found in the major organs of all test mice. Therefore, the LD ₅₀ value after single oral administration to the mouse of the mushroom extract according to the present invention is considered to be 2,000 mg / kg body weight or more for both males and females, and the mushroom extract according to the present invention is safe. Was confirmed to be high.

  The therapeutic agent for polycystic ovary syndrome (PCOS) according to the present invention is safe, has few side effects, and has an excellent effect of improving anovulation, by using an extract of mushrooms as an active ingredient. It can be applied safely to unmarried and young women as well as women who want to.

Claims (2)

A therapeutic agent for polycystic ovary syndrome (PCOS) comprising an extract of maitake as an active ingredient. The maitake extract is
(1) treating the maitake raw material with ethanol having a concentration of 90% or more to remove ethanol-soluble components;
(2) Thereafter, hot water extraction is performed, ethanol is added to the obtained hot water extract until the final volume concentration is 50 to 75%, the insoluble components produced are removed, and a supernatant is obtained,
(3) The therapeutic agent for polycystic ovary syndrome (PCOS) according to claim 1 , which is produced by a process comprising a step of collecting a fraction having a molecular weight of 14,000 or more from the obtained supernatant. .

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