JP6981641B2 - PDE5 activity inhibitor - Google Patents

Description

The present invention relates to a PDE5 activity inhibitor using a specific plant as a raw material and its use. More specifically, the present invention relates to a PDE5 activity inhibitor comprising a plant belonging to the genus False goat's beetle such as red pepper and an oral composition using the same.

In recent years, the number of patients with penile erectile dysfunction (ED) has increased along with the increase in the middle-aged and elderly population. An epidemiological survey conducted in 1998 reported that the number of ED patients in Japan was estimated to be 11.3 million, and one in three men aged 40 and over was one (Non-Patent Document 1). Decreased corpus cavernosum blood inflow due to sympathetic nerve activation due to stress, decreased corpus cavernosum blood inflow due to vascular disorders due to aging or lifestyle diseases, decreased guanylate cyclase activity, increased phosphodiesterase 5 (PDE5) activity, etc. Or, since it is said that ED develops when these occur in combination, it is considered that the number of ED patients will continue to increase as the elderly population increases.

Penile erection occurs when the smooth muscle of the corpus cavernosum relaxes and blood accumulates (hyperemia) in the corpus cavernosum sinus. NO nerves using nitric oxide (NO) as a neurotransmitter are distributed in the smooth muscle of the corpus cavernosum, and NO is released by sexual stimulation. The released NO activates guanylate cyclase in smooth muscle cells and increases cGMP, resulting in relaxation of smooth muscle and erection.

In order to improve ED, a substance having an action of inhibiting PDE5 activity (hereinafter, may be simply referred to as PDE5 inhibition) is used. Eleven families of PDEs have been identified from 1 to 11, and are enzymes that regulate intracellular signal transduction by hydrolyzing cAMP and cGMP. Among them, PDE5 is an enzyme specific to cGMP, which is abundant in the corpus cavernosum penis. By suppressing this inactivating action of cGMP, ED is improved by preventing the decomposition of cGMP increased by NO and increasing the cGMP concentration, relaxing the corpus cavernosum smooth muscle and increasing the local blood flow. Is possible.

PDE5 inhibitors currently in use include sildenafil, vardenafil, and tadalafil, but their side effects include dizziness due to decreased blood pressure, headache and facial burning, and cyanopsia due to PDE6 inhibition, especially angina. It is contraindicated for people taking nitrates or NO donors for the treatment of severe cardiovascular disorders, etc., as they may die from a sudden drop in blood pressure. Regarding the incidence of side effects of these PDE5 inhibitors, Viagra (sildenafil) is 30-40% at single doses of 25 mg, 50 mg, and 100 mg, and Cialis (tadalafil) is about 30% at 2.5-20 mg. , And Levitra (Vardenafil) are reported to be about 30% at 5 to 20 mg (Non-Patent Documents 2 to 4).

Food ingredients having a PDE5 inhibitory effect have also been studied so far, and 5,7-dimethoxyflavone contained in black ginger (Non-Patent Document 5), icariin contained in Epimedium (Non-Patent Document 6), etc. have been reported. ing.

Oral ingestions in which food ingredients such as trehalose (Patent Document 1), vitamin C and vitamin P (Patent Document 2), and arginine (Patent Document 3) are combined with a PDE5 inhibitor have been proposed. There are no reports that it has a PDE5 inhibitory effect alone. Further, a male function improving agent by increasing NO in vascular endothelial cells such as a nourishing tonic beverage containing a high amount of arginine (Patent Document 4) and an egg white hydrolyzate (Patent Document 5) has been proposed. Since cell-derived NO is considered to be involved in the maintenance of penile erection (Non-Patent Document 7), and the above-mentioned nitrates and NO donors are not used as ED therapeutic agents, endothelial cell-derived NO is directly used. It is not considered to be involved in the prevention or improvement of ED. On the other hand, since PDE5 is involved in penile erection and its maintenance, inhibition of PDE5 is very important for prevention and improvement of ED.

Japanese Unexamined Patent Publication No. 2012-197274 Japanese Unexamined Patent Publication No. 2012-413337 Japanese Unexamined Patent Publication No. 2010-163426 Japanese Unexamined Patent Publication No. 2007-116939 Japanese Unexamined Patent Publication No. 2011-173817

Eiji Marui, Epidemiology and Risk Factors of ED in Japan, 2002, History of Medicine, Vol. 201, pp. 397-400 Viagra Interview Form, 14th Edition, Pfizer, 2016 Cialis Interview Form, 5th Edition, Nippon Shinyaku, 2015 Levitra Interview Form, 14th Edition, Bayer Yakuhin, 2016 Prapapan Temkitthawon et al., "Kaempferia parviflora, a plant used in traditional medicine to enhance sexual permanent permanent Ethnopharmacol. , Vol. 137, pp. 1437 to pp. 1441 Hongxiu Ning et al., "Effects of icariin on phosphodiesterase-5 activity in vitro and cyclic guanosine gnosine monophosphate levelin car Haruaki Sasaki et al., Male Dysfunction, 2016, Showa Bachelor's Journal, Vol. 76, pp. 133-139

Although the drug has a PDE5 inhibitory effect, it has side effects, and there is a limitation that the dosage should be adhered to under the guidance of a doctor. In addition, the above-mentioned ingredients separated from foods are practically ineffective, are based on experimental results under impractical intake conditions, or must be ingested in large amounts in a normal dietary form. None of them could exert a sufficiently satisfactory effect.
In view of this situation, it is an object of the present invention to develop a PDE5 activity inhibitor derived from a natural product that can be effectively used for the prevention and improvement of ED, and to provide it as a composition in a mode that can be effectively utilized industrially. did.

In order to solve the above-mentioned problems, the present inventors have diligently studied various materials having an action of suppressing PDE5 activity, and as a result, plants belonging to the genus False goat's beetle such as red pepper can suppress PDE5 activity extremely strongly. In addition, they have found that this can be effectively used as an oral composition for foods and drinks, and have completed the present invention.

That is, the feature of the present invention is a PDE5 activity inhibitor containing a plant belonging to the genus False goat's beetle as an active ingredient. In this PDE5 activity inhibitor, it is desirable that the plant belonging to the genus False goat's beetle is false goat's beetle.

It is more desirable that the PDE5 activity inhibitor contains an extract or a purified product thereof obtained by extracting a plant belonging to the genus False goat's beetle such as red pepper with water and / or a hydrophilic organic solvent.

It is more desirable that the PDE5 activity inhibitor is used as an extract obtained by extracting a plant belonging to the genus False goat's beetle such as red pepper with water and / or a hydrophilic organic solvent.

Another feature of the present invention is an oral composition comprising the PDE5 activity inhibitor. It is desirable that this oral composition be a food or drink.

Another feature of the present invention is that the PDE5 activity inhibitor or oral composition is for preventing ED.

A further other feature of the present invention lies in a method for producing a PDE5 activity inhibitor containing an extract obtained by extracting a plant belonging to the genus False goat's beetle, preferably red pepper, with water and / or a hydrophilic organic solvent as an active ingredient. ..

Yet another feature of the present invention is mediated by inhibition of PDE5 activity, characterized by oral administration of an extract obtained by extracting a plant belonging to the genus False goat's beetle, preferably red pepper, with water and / or a hydrophilic organic solvent. There is a way to prevent the occurrence of ED.

Yet another feature of the present invention is a method of orally administering or ingesting the PDE5 activity inhibitor or oral composition to prevent ED.

Plants belonging to the genus False goat's beetle such as Astilbe odontophyllum and Astilbe odontophyllum according to the present invention, and extracts thereof, particularly the extracts containing hydrophilic components, are produced by oral ingestion thereof with daily lifestyle and aging. It has the effect of significantly preventing ED.
Further, according to the present invention, there are no side effects that have been a problem with conventional ED therapeutic agents, no guidance from a doctor or the like is required, and an appropriate amount can be effectively used for prevention or improvement of ED, which brings about a certain effect. It has become possible to provide a PDE5 activity inhibitor derived from a natural product.

The present invention will be described in detail below. First, the PDE5 activity inhibitor of the present invention contains a plant belonging to the genus False goat's beetle (Astilbe) or an extract thereof as an active ingredient. Plants belonging to the genus False goat's beetle are classified into the family Saxifragaceae, and there are various plants according to the present invention, and as a representative example thereof, red pepper (scientific name: Astilbe sumbergii (SIEB. Et ZUCC.) MIQ.) Can be mentioned. Akashoma is a perennial plant that grows naturally in the mountains of Japan, and its rhizome is called red buttercup. It has been. In the present invention, what is called red or red hemp is also included.

As an example of a plant belonging to the genus False goat's beetle, Astilbe chinensis, A. et al. austrosinensis, A. et al. thunbergii, A.M. thumbergii (SIEB.et ZUCC.) Miq. : Akashouma, A. thumbergii (SIEB.et ZUCC.) MIQ. var. congesta BOISS. (= A. odontophylla MIQ.): Astilbe odontophylla, A. polyandra, A. grandis, A. rivularis, A. japonica (MORR. et DECNE.) A. GRAY: Buck's beetle, A. microphylla KNOLL: False goat's beetle, A. Myriantha and the like can be mentioned. In the present invention, Akashouma and Astilbe odontophyllum are suitable raw materials, and Akashouma is the most desirable.

In the present invention, as a plant belonging to the genus False goat's beetle such as red pepper used as a raw material, a raw plant body or a dried product thereof can be used, but the site is preferably root and / or rhizome, and the embodiment is root and /. Alternatively, it is desirable that the rhizome is appropriately processed by drying, shredding or crushing.

In the extraction treatment, it is preferable to use water, a hydrophilic organic solvent, or a mixed solvent thereof as the extraction solvent. Lower monohydric alcohols such as methanol, ethanol, propanol and isopropanol, polyhydric alcohols such as propylene glycol, 1,3-butanediol and glycerin, acetone, methyl ethyl ketone, ether, petroleum ether, ethyl acetate and These hydrous substances and mixtures can be exemplified. In order to efficiently obtain an extract for achieving the desired effect of the present invention, it is desirable to use ethanol, acetone, and a water-containing substance thereof as an extraction solvent. The water content of the water-containing material is, for example, 1 to 99% by mass, more preferably 10 to 50% by mass in the case of ethanol, and 1 to 50% by mass, more preferably 10 to 30 in the case of acetone. It is mass%. Outside these ranges, the desired effect of the present invention is reduced or the yield of the extract is reduced. Further, in order to easily and efficiently obtain the purified product according to the present invention, for example, it is preferable to extract with hydrous ethanol or hydrous acetone and collect the extract.

In the extraction treatment, an extraction solvent of about 1 to 100 times by mass is added to the raw material, and the mixture is appropriately stirred under normal pressure or pressure at room temperature or in a heated state for 10 minutes to several days. Then, the insoluble material can be filtered or centrifuged to obtain an extract solution according to the present invention, and the present invention is further removed by removing the solvent from the extract solution by known means such as distillation under reduced pressure, spray drying, and freeze drying. The extract according to the above can be obtained. Further, a purified product is obtained by subjecting this extract to a known purification treatment such as fractionation with an adsorbent (silica gel, magnesium silicate, ion exchange resin, activated alumina, cellulose, activated carbon, etc.), solvent separation, and a combination thereof. Any treated product can be used in the present invention, but the above-mentioned extract or a purified product thereof is desirable from the viewpoint of convenience of use described later and the effect of the present invention.

The product obtained by treating the raw material according to the present invention as described above is a complex composition containing a wide variety of components, although there are quantitative differences, and it is a complex composition containing bergenin, astilbin, taxifolin, polyphenols, and tannins. It contains various water-soluble components (polysaccharides, oligosaccharides, monosaccharides, proteins, peptides, amino acids, complexes thereof, etc.).

The PDE5 inhibitor of the present invention uses the treated product of a plant belonging to the genus Chidake as an active substance, and if necessary, known excipients, binders, disintegrants, abundant agents, diluents and the like, for example, saccharides. , Sugar alcohols, starches, celluloses, dicalcium phosphate, mica, talc, purified water, ethanol and the like can be mixed in appropriate amounts and processed into solid, powder, paste or liquid forms. Further, a known substance or extract having a PDE5 inhibitory action as described above may be appropriately used in combination. The content of the treated product of the false goat's beetle plant in the PDE5 inhibitor of the present invention is difficult to uniformly define due to the difference in its morphology and the like, but is about 0.1% by mass to 100% by mass, more preferably about 10. It is from% by mass to about 80% by mass. If it is less than about 0.1% by mass, the desired effect may not be exhibited in the PDE5 inhibitor or oral composition of the present invention.

Next, in the oral composition of the present invention, the PDE5 inhibitor may be used as it is as an oral composition, but it is known to be appropriately used for ordinary foods and drinks, pharmaceuticals, quasi-drugs, animal feeds and the like. It is desirable to use the above additives in combination and add them by a conventional method to form a composition. Here, the known additive may be any additive that can be blended in the above-mentioned various products and does not contradict the gist of the present invention, and is, for example, an excipient, a binder, a disintegrant, and a lubricant. , Wetting agents, fluidizing agents, preservatives, surfactants, stabilizers, diluents, solubilizers, tonicity agents, bactericides, preservatives, flavoring agents, deodorants, coloring agents, fragrances and other additives Can be used.

In the present invention, the PDE5 inhibitor can be used as it is in various products in foods and drinks, pharmaceuticals, quasi-drugs, feeds, and other industrial fields, or a part of the compounding raw materials of such products. It can also be used in the mode of using as. In particular, it is preferable to use it as an oral composition for preventing the above-mentioned ED, and the most preferable embodiment of this oral composition is food and drink. This example will be described below, but the present invention is not limited thereto.

Specific examples of foods and drinks include vegetable juices, fruit juice beverages, refreshing beverages, tea and other beverages, soups, jellies, puddings, yogurts, cake premix products, confectionery, sprinkles, miso, soy sauce, sauces, dressings, and mayonnaise. , Vegetable cream, seasonings such as grilled meat sauce and noodle soup, noodles, udon, soba, spaghetti, processed meat and fish meat such as ham and sausage, hamburger, croquette, sprinkle, boiled noodles, jam, milk, cream, butter, Powdered, solid or liquid dairy products such as spreads and cheese, various general processed foods such as margarine, bread, cakes, cookies, chocolates, candies, gummy and gum, as well as powdered, granular and rounded foods. Examples thereof include tablet-shaped, soft-capsule-shaped, hard-capsule-shaped, pasty-shaped or liquid-shaped nutritional supplements, foods for specified health use, functional foods, health foods, concentrated liquid foods, and therapeutic foods for swallowing disorders. Of these, in order to be able to blend the active ingredient according to the present invention in a high concentration and to realize the effect of the present invention more reliably, dietary supplements such as powder, granule, tablet, capsule, and liquid , Foods for specified health use, functional foods or health foods are desirable.

In order to produce these foods and drinks, the PDE5 inhibitor of the present invention and known raw materials may be used, or a part of the known raw materials may be replaced with the PDE5 inhibitor of the present invention and produced by a conventional method. For example, the PDE5 inhibitor of the present invention can be used as necessary for glucose (dextrose), textrin, lactose, starch or processed products thereof, excipients such as cellulose powder, vitamins, minerals, fats and oils of animals, plants and seafood, and proteins. (Proteins derived from animals and plants and yeast, their hydrolyzates, etc.), sugars, pigments, fragrances, antioxidants, surfactants, other edible additives, powders and extracts containing various nutritional functional ingredients, etc. It can be mixed with and processed into the shape of powder, granules, pellets, tablets, etc., or processed into the general processed food of the above example by a conventional method, and the liquid material mixed with these can be processed into gelatin, sodium alginate, carboxymethyl cellulose, etc. It is suitable to coat it with a coating agent and form it into a soft capsule, or to process it into a beverage (drinks) and use it as a dietary supplement or a health food.

The content of the PDE5 inhibitor of the present invention in such foods and drinks is uniform depending on the form of the food and drink, the form of the PDE5 inhibitor of the present invention, the content of the active ingredient, the type, the component, the blending amount and the like of the compounded raw material to be used in combination. Although it is difficult to specify in the above, it is about 0.01% by mass to about 90% by mass, more preferably about, in terms of the extract according to the present invention (extract extracted with water and / or a hydrophilic organic solvent of a plant belonging to the genus Chidakesashi). It is 1% by mass to about 50% by mass. For foods and drinks less than about 0.01% by mass, a large amount of the foods and drinks must be ingested in order to expect the desired effect of the extract, while when the content of the extract exceeds about 90% by mass. It may be difficult to produce practical food and drink.

In using the food and drink of the present invention, in the case of a human adult, the standard daily intake of the extract is about 10 mg to about 1,000 mg, preferably about 30 mg to about 500 mg, and more preferably about 50 mg to about 300 mg. It can be taken into the body by any method, for example, at the same time as or before or after ingesting a meal, by oral ingestion, tube administration, or the like.

Next, the present invention will be described in detail with reference to examples, but the present invention is not limited thereto. In each example,%, parts and ratios are all based on mass.

Production Example 1
The rhizome of False goat's beetle (SIEB. EtZUCC.) MIQ. Was washed with water, chopped into 1 to 2 cm in length and 3 to 5 mm in width, and then dried in the sun to prepare a dried false goat's beetle. 1 kg of this dried red pepper was charged in a stainless steel extraction kettle, 9 L of water was added, and extraction treatment was performed at 70 ° C. for 6 hours with appropriate stirring. Then, the insoluble residue was filtered off to collect an extract, and the extract was subjected to vacuum concentration, freeze-drying and pulverization to obtain a reddish brown extract powder (Sample A).

Manufacturing example 2
1 kg of the dried red pepper prepared in Production Example 1 was placed in a stainless steel extraction kettle, 9 L of hydrous ethanol having a water content of 45% was added, and extraction treatment was performed at 70 ° C. for 6 hours with appropriate stirring. Then, the insoluble residue was filtered off to collect an extract, and the extract was subjected to vacuum concentration, freeze-drying and pulverization to obtain a reddish brown extract powder (Sample B).

Production example 3
1 kg of the dried red pepper prepared in Production Example 1 was placed in a stainless steel extraction kettle, 9 L of ethanol was added, and extraction treatment was performed at 70 ° C. for 6 hours with appropriate stirring. Then, the insoluble residue was filtered off to collect an extract, and the extract was subjected to vacuum concentration, freeze-drying and pulverization to obtain a light reddish brown extract powder (Sample C).

Test Example The PDE5 activity inhibitory action of each sample obtained in the above-mentioned production example and the safety of the red pepper extract powder were investigated by the methods described below.

Test Example 1: PDE5 activity inhibitory action by red pepper extract powder (1)
The PDE5 activity inhibitory action was carried out using PDE5A (human), (Recombinant) (manufactured by Enzo Biochem) as enzymes, and according to the Cyclic Nucleotide Phosphodiesterase assay kit (manufactured by Enzo Biochem). Samples A, B and C were dissolved in Assay Buffer to a final concentration of 1,10 μg / mL. 10 μL of sample or 40 μL of standard solution was added to the 96-well half area plate included in the kit. 20 μL of cGMP substrate (0.5 mM), 5 μL of PDE5A and Assay Buffer were added to the wells to which the sample was added to make the total volume 40 μL. 10 μL of 5'-nuleotidase was added to all wells and incubated at 30 ° C. for 30 minutes. 100 μL of BIOMOL GREEN Reagent was added, and the mixture was allowed to stand at room temperature for 30 minutes, and the absorbance at 620 nm was measured. A calibration curve was prepared from the standard product, and the 5'-GMP concentration produced by PDE5 was determined. The PDE5 activity inhibition rate of each sample is calculated by the following formula, that is, [{(control 5'-GMP concentration)-(sample-added sample 5'-GMP concentration)} / (control 5'-) based on the absorbance. It was calculated from GMP concentration)] × 100, and was shown as a relative value when 0% was used when no sample was added (control), which was carried out at the same time (Tables 1 to 3).

The results are shown in Table 1. From the data in Table 1, it was confirmed that the water of the red pepper extract powder or the hydrous ethanol extract has a PDE5 inhibitory effect.

Test Example 2: PDE5 activity inhibitory action by red pepper extract powder (2)
It was investigated whether the red pepper extract powder inhibits PDE5 activity in a concentration-dependent manner. The PDE5 activity was measured using the same method as in Test Example 1. The sample B was dissolved in Assay Buffer so as to have a final concentration of 1, 2, 3, 4, 5, and 10 μg / mL.

The results are shown in Table 2. From the data in Table 2, it was confirmed that the red pepper extract powder according to the present invention inhibits PDE5 activity in a concentration-dependent manner. Furthermore, when the 50% inhibition rate was calculated based on this data, it was 1.78 μg / mL, and it was confirmed that the red pepper extract powder had a very strong PDE5 inhibitory effect.

Test Example 3: PDE5 activity inhibitory effect of red pepper extract powder and other ED improving agents In this test, as a male function improving agent compared with red pepper extract powder, maca extract powder (MACAXS (registered trademark) -HC, manufactured by TOWA CORPORATION). (Target ▲ 1 ▼), BHN black turmeric extract powder (manufactured by BHN, containing 2% 5,7-dimethoxyflavone) (target ▲ 2 ▼) and sildenafil (manufactured by Cayman Chemical) (target ▲ 3 ▼) was used. The PDE5 activity was measured using the same method as in Test Example 1. The sample B, maca extract powder and black turmeric extract powder were dissolved in Assay Buffer so that the final concentration was 1,10 μg / mL, and sildenafil was 0.01, 0.1 μg / mL. used.

The results are shown in Table 3. From the data in Table 3, the red pepper extract powder according to the present invention showed a very strong PDE5 inhibition rate as compared with the maca extract powder and the black turmeric extract powder known as male function improving agents. It was also confirmed that the red pepper extract powder has an inhibition rate of about 1/10 of sildenafil used as a PDE5 inhibitor.

Test Example 4 Safety of red pepper extract powder (blood pressure)
The subjects were healthy males (age: 32 to 59 years old), the number of subjects was 5 (2 obese, 1 diabetic, 2 standard weights), and the test meal was a hard capsule (red pepper extract powder 200 mg / tablet prescription). Three capsules a day (before each meal) were ingested for 12 weeks, and the blood pressure status of the subjects was examined. Table 4 shows the data of the subjects, and Table 5 shows the data showing the blood pressure status of the subjects.

Blood pressure was stable in all subjects during the study, and no dizziness, headache, or hot flashes characteristic of the side effects of currently used PDE5 inhibitors (sildenafil, vardenafil, tadalafil, etc.) were observed. .. No abnormal findings such as cyanopsia were observed.

Prototype example 1
The sample A or the sample B was subjected to a capsule filling machine, and a gelatin-coated hard capsule preparation having a content of 250 mg per capsule was prototyped by a conventional method. This capsule formulation can be used as an orally ingestible drug or dietary supplement.

Prototype example 2
Sample A or sample B 5 g was added to 1 L of commercially available vegetable juice and mixed to prepare a prototype vegetable juice for enhancing male function. This was comparable to the original vegetable juice.

Prototype example 3
Sample A or Sample B 5.0 kg, modified starch (manufactured by Matsutani Chemical Co., Ltd., trade name: Pineflow (registered trademark)) 3.5 kg, tertiary calcium phosphate 0.3 kg, vitamin B 10.3 kg, vitamin B 20.3 kg, vitamin It was charged into a compounding machine together with 60.2 kg of B and 0.4 kg of vitamin C, and stirred and mixed for 10 minutes. The mixture was supplied to a direct-type tableting machine to prepare a tablet having a diameter of 7 mm, a height of 4 mm, and a weight of 150 mg, and then a shellac thin film was coated with a coating machine to make a tablet-shaped food product as a prototype.

By orally ingesting plants belonging to the genus False goat's beetle, such as Akashouma and Astilbe odontophyllum, and the extracts thereof, especially those containing hydrophilic components, ED caused by daily lifestyle and aging is adversely affected. It can have a remarkable preventive effect without it.
Further, the present invention can be effectively used as foods and drinks containing the extract, pharmaceuticals, quasi-drugs, feeds, and various other products in the industrial field.

Claims (4)

A PDE5 activity inhibitor comprising water and / or a hydrophilic organic solvent extract or a purified product thereof of False goat's beetle, which is a plant belonging to the genus False goat's beetle. The PDE5 activity inhibitor according to claim 1, which is for preventing ED. A method for producing a PDE5 activity inhibitor, which comprises containing an extract obtained by extracting red pepper, which is a plant belonging to the genus False goat's beetle, with water and / or a hydrophilic organic solvent as an active ingredient. A method of orally administering or ingesting the PDE5 activity inhibitor according to claim 1 to prevent ED (excluding the method of treating humans) .