PDE5 Inhibitors

Abstract

To provide a new substance having a PDE5 inhibitory effect.SOLUTION: The PDE5 inhibitor includes a powder of Paecilomyces tenuipes that parasitizes silkworms, or an extract thereof.SELECTED DRAWING: None

Description

[Technical field]

[0001] The technology relates to phosphodiesterase (PDE) 5 inhibitors. [Background technology]

[0002] Substances having PDE5 inhibitory activity are known to be effective in preventing or improving urinary tract disorders and pulmonary arterial hypertension (Non-Patent Document 1). Naturally derived substances having PDE5 inhibitory activity are also known (Patent Documents 1 and 2). [Prior art documents] [Patent documents]

[0003] [Patent Document 1] Patent Publication No. 2019-34920 [Patent Document 2] Patent Publication 2018-154610 [Non-patent literature]

[0004] [Non-Patent Document 1] Japanese Journal of Pharmacology, 154, 265-269, 2019 Summary of the Invention [Problem to be solved by the invention]

[0005] One objective is to provide a new substance that has PDE5 inhibitory activity. [Means for solving the problem]

[0006] As a result of intensive research aimed at solving the above problems, it was found that Bombyx mori has a PDE5 inhibitory effect. Based on this finding, further research was carried out, and the following representative inventions are provided. Item 1 A PDE5 inhibitor comprising a powder or extract of Silkworm Cordyceps gracilis. Section 2 Item 1. The PDE5 inhibitor according to item 1, which is in the form of a food or drink composition. Section 3 Item 3. The PDE5 inhibitor according to item 1 or 2, for use in preventing or ameliorating dysuria and / or pulmonary arterial hypertension. Section 4 Use of a powder or extract of Polyporus nerka for producing a PDE5 inhibitor. Section 5 Use of a powder or extract of Polyporus nervosa for producing a composition for preventing or improving dysuria and / or pulmonary arterial hypertension. Section 6 A method of administering a powder or extract of Silkworm Polyporus nigricans to a subject in need of PDE5 inhibition. Section 7 Item 7. The method according to item 6, wherein the subject is in need of prevention or amelioration of dysuria and / or pulmonary arterial hypertension. Section 8 A powder or extract of Polyporus nervosa for use in inhibiting PDE5. Section 9 A powder or extract of Polyporus nervosa for use in the prevention or improvement of dysuria and / or pulmonary arterial hypertension. Effect of the Invention

[0007] Provided is a substance having a PDE5 inhibitory effect. In one embodiment, a means for preventing or ameliorating dysuria and / or pulmonary arterial hypertension utilizing a PDE5 inhibitory effect is provided. [Brief description of the drawings]

[0008] [Figure 1] The results of measuring the PDE5 inhibitory activity of Bombyx mori. "Sample" indicates a hot water extract of Bombyx mori. "Substrate" indicates cGMP. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0009] The PDE5 inhibitor preferably contains Bombyx mori or an extract thereof. Bombyx mori is P. tenuipes, also called "Cordyceps sinensis", which grows parasitically on silkworms. Bombyx mori can be either commercially available or cultivated. Any method for cultivating Bombyx mori may be used, and for example, a method may be used in which silkworm larvae or silkworm pupae are heat sterilized, and then Cordyceps sinensis bacteria are inoculated and cultivated. In addition, a method disclosed in JP 2005-237201 A may be used. Specifically, silkworm larvae or silkworm pupae before they form cocoons are boiled and dried, and 50 to 90 weight percent of this dried silkworm powder is mixed with a dried food powder consisting of one or more types of dried powder of beans, grains, seaweed, or mushrooms as the remainder. Culture liquid is added to this and kneaded, and this is spread on the bottom of a cultivation box to create a medium. This medium is then sealed in an inoculation bag and heat sterilized, after which the medium is inoculated with Cordyceps sinensis bacteria for cultivation.

[0010] In one embodiment, the silkworm Polyporus is preferably in the form of a powder. The silkworm Polyporus powder can be obtained by freeze-drying and pulverizing the silkworm Polyporus. The silkworm Polyporus powder may be obtained by powdering only the fruiting body of the silkworm Polyporus, but is preferably obtained by powdering the fruiting body and the host (e.g., silkworm).

[0011] In one embodiment, the silkworm cordifolia is preferably in the form of an extract. The method and conditions used to obtain the extract are arbitrary. For example, the silkworm cordifolia may be cut, crushed and / or dried as is, and squeezed or extracted using a suitable solvent. The type of solvent is not particularly limited, and examples thereof include water, lower alcohols such as methanol, ethanol, and isopropanol, polyhydric alcohols such as propylene glycol and 1,3-butylene glycol, ketones such as acetone, esters such as diethyl ether, dioxane, acetonitrile, and ethyl acetate, dimethyl sulfoxide, xylene, benzene, and chloroform. In one embodiment, the preferred solvent is water, and preferably hot water (60° C. or higher, 70° C. or higher, 80° C. or higher, or 90° C. or higher).

[0012] In one embodiment, the extract of Bombyx mori is preferably obtained by the following method. That is, hot water is added to the dried powder of Bombyx mori, the mixture is stirred at a temperature of 60°C or more and 80°C or less for a predetermined time (for example, 1 hour or more and 5 hours or less), and the filtrate obtained by filtration is dried to obtain the extract. The mixing ratio of the dry powder and the hot water is arbitrary, but can be, for example, 5 to 20 parts by weight or 7 to 15 parts by weight per 1 part by weight of the dry powder.

[0013] The moisture content of the powder or extract of Bombyx mori is not particularly limited, but can be, for example, 20% by weight or less, 10% by weight or less, 8% by weight or less, or 5% by weight or less.

[0014] The PDE5 inhibitor may be composed of only the powder of the silkworm Cordyceps or its extract, or may further contain any other components.The other components may include, for example, excipients, binders, disintegrants, lubricants, wetting agents, flow agents, preservatives, surfactants, stabilizers, diluents, dissolving agents, isotonicity agents, bactericides, preservatives, flavoring agents, odorants, coloring agents, flavoring agents, and other fragrances.Other components may also be selected from other components having PDE5 inhibitory activity or other physiologically active components.The form of the PDE5 inhibitor is arbitrary, and may be, for example, powder, granules, tablets, capsules, or liquid.

[0015] The amount of the silkworm larvae or its extract contained in the PDE5 inhibitor is arbitrary, and can be set within the range of 0.1% or more and 100% or less in terms of dry weight.The lower limit of the content can be, for example, 1% or more, 2% or more, 5% or more, 10% or more, 15% or more, 20% or more, 30% or more, 40% or more, 50% or more, 60% or more or 70% or more, and the upper limit can be 99% or less, 95% or less, 90% or less, 80% or less, 70% or less, or 60% or less.These lower and upper limits can be combined arbitrarily.

[0016] In one embodiment, the PDE5 inhibitor is preferably in the form of a food or drink. Food or drink includes not only general foods, but also health functional foods (specifically, foods for specified health uses, foods with nutritional functions, and foods with functional claims), foods for the sick, and supplements.

[0017] In one embodiment, the PDE5 inhibitor may be in the form of a liquid such as a solution, emulsion, suspension, or syrup, or in the form of a semi-solid or solid such as a powder, granules, fine granules, tablet, pill, capsule (including hard capsules and soft capsules), troche, chewable, gel, cream, paste, mousse, or sheet.

[0018] General foods are not particularly limited, and examples thereof include beverages (e.g., fruit juice drinks, vegetable juices, soft drinks, alcoholic drinks, carbonated drinks, sports drinks, tonic drinks, nutritional drinks, etc.), sweets (e.g., snacks, rice crackers, chocolates, gummies, candies, jellies, puddings, Japanese sweets, Western sweets, etc.), seasonings (e.g., soy sauce, miso, mirin-style seasoning, mixed spices, sauces, soups, sauces, ketchup, etc.), dressings, noodles (e.g., udon, soba, spaghetti, etc.), processed meat and fish foods (e.g., sausages, wieners, ham, hamburger steaks, etc.), dairy products (yogurt, milk, cheese, cream, butter, etc.), etc.

[0019] In one embodiment, the above-mentioned PDE5 inhibitor can be used to prevent or improve dysuria and / or pulmonary arterial hypertension by inhibiting the activity of PDE5. The cause and type of urinary tract disorder can be any, but examples of such disorders include urinary tract disorders accompanied by lower urinary tract symptoms.

[0020] In one embodiment, the PDE5 inhibitor can be in the form of a medicine. A medicine containing a PDE5 inhibitor can be prepared by combining a powder or extract of Polyporus nigricans with a pharmacopoeias and a pharmacopoeias acceptable excipient as necessary.

[0021] The intake (administration) amount of the PDE5 inhibitor is not particularly limited as long as the desired effect is achieved, and can be appropriately set according to the physique, age, symptoms, application form, purpose of use, etc. of the subject (subject animal). For example, the intake (administration) amount per day can be set in the range of 0.001 to 8000 mg, 0.01 to 5000 mg, etc., calculated as the dry weight of the silkworm Cordyceps or its extract, based on an adult weighing 60 kg. The number of intakes (administrations) is arbitrary, and may be, for example, a single intake (administration) or multiple intakes (administrations) per day.

[0022] As mentioned above, since Bombyx mori or its extract has PDE5 inhibitory effect, Bombyx mori powder or extract can be used to manufacture PDE5 inhibitor. In addition, Bombyx mori powder or extract can be used to manufacture a composition for preventing or improving dysuria and / or pulmonary arterial hypertension.

[0023] As mentioned above, since Bombyx mori or its extract has PDE5 inhibitory effect, Bombyx mori powder or extract can be administered to a subject who needs PDE5 inhibition to provide PDE5 inhibitory effect.In addition, since PDE5 inhibition can prevent or improve dysuria and / or pulmonary arterial hypertension, Bombyx mori powder or extract can be administered to a subject who needs to prevent or improve dysuria and / or pulmonary arterial hypertension to prevent or improve said condition. EXAMPLES

[0024] The present invention will be described in more detail below with reference to examples, but the present invention is not limited to these examples.

[0025] 1. Silkworm Cordyceps nigricans hot water extract Silkworm Polyps (Bombyx mori) obtained from silkworm pupae was freeze-dried and pulverized in a mixer to obtain silkworm Polyps powder. The resulting powder (42 g) and 420 mL of ultrapure water were placed in a reaction vessel and stirred at 70°C for 3 hours, after which the residue was removed by filtration to obtain a solution. The solution was then concentrated to 30 mL, and spray-dried to obtain a silkworm Polyps hot water extract.

[0026] 2. Measurement of PDE5 inhibitory activity PDE5 inhibitory activity was measured using PDE-Glo™ Phosphodiesterase Assay (Promega). The samples used were prepared by diluting the hot water extract of Bombyx mori with ultrapure water to final concentrations of 33 μg / ml, 11 μg / g, 3.7 μg / ml, and 1.2 μg / ml. The enzyme used was phosphodiesterase 5A1 human (Sigma-Aldrich). 3100 U / μl of PDE1μ was mixed with 205.7 μl of reaction buffer to prepare a PDE solution. 15 μl of PDE solution was mixed with 10 μl of each sample, spun down for 2 minutes, and shaken at room temperature for 5 minutes. 25 μl of this mixture was mixed with 25 μl of 20 μM cGMP, and 8 μl was dispensed into PCR tubes. This was placed on a shaker and reacted at room temperature at 400 rpm. 4 μl of detection buffer was added to each tube, and after spinning down, the tubes were shaken at room temperature for 20 minutes. 10 μl of this and Kinase-Glo reagent were dispensed into each well of a 384-well plate (Thermo Fisher), and the plates were shaken at room temperature to react. The relative inhibition rate of the Bombyx mori hot water extract was measured using a test example in which a sample without the addition of the Bombyx mori hot water extract was used as the 0% control, and a test example in which no PDE was added was used as the 100% control. The measurement was repeated four times. The results are shown in Figure 1.

[0027] From the results in Figure 1, it was confirmed that the hot water extract of Bombyx mori has PDE5 inhibitory activity and that this activity increases in a concentration-dependent manner. Based on these results, Bombyx mori is considered to be effective in treating or preventing diseases such as pulmonary arterial hypertension and dysuria.

Claims

1. A PDE5 inhibitor comprising a powder or extract of Silkworm Polyporus arbutus as an active ingredient, said extract being an extract made from water or a lower alcohol.

2. The PDE5 inhibitor according to claim 1 , which is in the form of a food or beverage composition.

3. The PDE5 inhibitor according to claim 1 or 2, which is for preventing or improving dysuria and / or pulmonary arterial hypertension.

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