JP6356492B2 - Anti-inflammatory agent - Google Patents

Description

  The present invention relates to an anti-inflammatory agent.

  Piceatannol is a stilbene compound, for example, contained in the seeds of passion fruit, a fruit of the genus Passiflora (Passiflora), and causes melanin that causes pigmentation due to spots, buckwheat, sunburn, etc. It has been reported that there is an effect of suppressing the generation of (see Patent Document 1).

  This piceatannol has a strong anti-inflammatory action and suppresses secretion of TNF-alpha and IL-8 in response to LPS stimulation in human peripheral blood mononuclear cells and the like (see Non-Patent Document 1). ).

JP 2009-102298 A

Richard N et al. Mol Nutr Food Res. 2005 vol.49, pp.431-442.

  An object of the present invention is to provide a novel anti-inflammatory agent.

  As a result of diligent efforts to develop a method for using passion fruit seed extract, the present inventors have found that passion fruit seed extract has a strong anti-inflammatory effect. And it discovered that isolapontigenin, which is one of the analogues of piceatannol, also has a strong anti-inflammatory action, leading to the completion of the present invention.

  One embodiment of the present invention is an anti-inflammatory agent, an IL-8 production inhibitor, or a TNF-α production inhibitor containing passion fruit seed extract or isolapontigenin as an active ingredient.

  According to the present invention, a novel anti-inflammatory agent can be provided.

In one Example of this invention, it is a graph showing the result of having measured the quantity of IL-8 and TNF- (alpha) which the human peripheral blood mononuclear cell secreted in response to LPS in the presence of passion fruit seed extract. In one Example of this invention, it is a graph showing the result of having measured the quantity of IL-8 and TNF- (alpha) which the human peripheral blood mononuclear cell secreted in response to LPS in the presence of isolapontigenin.

  The objects, features, advantages, and ideas of the present invention will be apparent to those skilled in the art from the description of the present specification, and those skilled in the art can easily reproduce the present invention from the description of the present specification. The embodiments and specific examples of the invention described below show preferred embodiments of the present invention, and are shown for illustration or explanation. It is not limited. It will be apparent to those skilled in the art that various modifications and variations can be made based on the description of the present specification within the spirit and scope of the present invention disclosed herein.

(1) Isolapontigenin isolapontigenin (3,4 ', 5-Trihydroxy-3'-methoxy-trans-stilbene;3,4',5-trihydroxy-3'-methoxy-trans-stilbene) It is a compound which has the following structural formula.

Isolapontigenin is a commercially available product that is chemically synthesized and can be easily produced and obtained, but it is contained in a large amount in the Indonesian native meremon (Gnetum gnemon), and natural isolapontigenin is isolated from these plants. May be.

(2) Production Method of Passion Fruit Seed Extract As a specific production method of the passion fruit seed extract, a known method can be used. For example, after the passion fruit seed is dried, it is crushed, pulverized, or cut. A seed decomposition product is obtained, extracted using a solvent, and an extract is obtained by removing the residue. Further, an extract can be obtained by removing the solvent from the extract. Any of these stages can be used as the passion fruit seed extract of the present invention.

The type of the solvent used for extraction can be appropriately selected by those skilled in the art. For example, water, methanol, ethanol, acetone, ethyl acetate, glycerin, propylene glycol, dipropylene glycol, 1,3-butylene glycol , Dimethyl sulfoxide, acetonitrile, N, N-dimethylformamide, 2-propanol, 1,4-dioxane, hexane, chloroform, dichloromethane, or a mixed solvent of two or more solvents selected from these, water , Ethanol, 1,3-butylene glycol, or a mixed solvent of two or more solvents selected from these, more preferably water, ethanol, or a mixed solvent of water and ethanol. When using a mixed solvent, the mixing ratio of each solvent is not particularly limited. For example, when using a mixed solvent of water and ethanol, the volume ratio of water to ethanol is 1:99 to 99: 1. The ratio is preferably 3:97 to 80:20, more preferably 5:95 to 50:50, and particularly preferably 10:90 to 40:60.

When water or a mixed solvent with water is used as the solvent, it is preferably hot water or a mixed solvent with hot water. Water or a mixed solvent with water may contain a salt, and may be a buffer (buffer) as an example of a solvent containing a salt. The pH of the buffer is not particularly limited, and may be acidic, neutral, or alkaline, but is preferably acidic, more preferably acidic at pH 6 or lower, and acidic from pH 1 to pH 5. More preferably. The kind of salt used for the buffer is not particularly limited, and examples thereof include citrate, malate, phosphate, acetate, and carbonate.

The method for removing the solvent from the extract is not particularly limited, and a known method can be used. For example, vacuum distillation, freeze drying, or spray drying (spray drying) may be used, but freeze drying or spray drying is preferable, and spray drying is more preferable.

The shape of the extract is not particularly limited, and may be, for example, a solid such as powder, an amorphous form, or an oil form.

(3) Anti-inflammatory agent and IL-8 production inhibitor, or TNF-α production inhibitor Passion fruit seed extract or isolapontigenin has an inhibitory effect on IL-8 or TNF-α production. Alternatively, it can be effectively used as a drug such as a TNF-α production inhibitor or an anti-inflammatory agent. The formulation of passion fruit seed extract or isolapontigenin is possible by a known method.

The drug according to the present invention may further contain various components generally used in addition to the active ingredient, if necessary, for example, one or more pharmaceutically acceptable excipients, disintegrants, Diluent, Lubricant, Flavor, Colorant, Sweetener, Flavoring Agent, Suspending Agent, Wetting Agent, Emulsifier, Dispersant, Auxiliary Agent, Preservative, Buffering Agent, Binder, Stabilizer, Coating Agent And so on.

As the administration route, either systemic administration or local administration can be selected. In this case, an appropriate administration route is selected according to the disease, symptoms and the like. The drug according to the present invention can be administered by either oral route or parenteral route. Examples of parenteral routes include normal intravenous administration and intraarterial administration, as well as subcutaneous, intradermal and intramuscular administration. Furthermore, transmucosal administration or transdermal administration can be performed.

When preparing an oral solid preparation, after adding excipients, binders, disintegrating agents, lubricants, coloring agents, corrigents, flavoring agents, etc. to the active ingredient, tablets by a conventional method, Coated tablets, granules, powders, capsules and the like can be produced. Such additives may be those commonly used in the art. For example, excipients include lactose, sucrose, sodium chloride, glucose, starch, calcium carbonate, kaolin, microcrystalline cellulose, silicic acid As a binder, water, ethanol, propanol, simple syrup, glucose solution, starch solution, gelatin solution, carboxymethylcellulose, hydroxypropylcellulose, hydroxypropyl starch, methylcellulose, ethylcellulose, shellac, calcium phosphate, polyvinylpyrrolidone, etc. Disintegrants include dry starch, sodium alginate, agar powder, sodium bicarbonate, calcium carbonate, sodium lauryl sulfate, stearic acid monoglyceride, lactose, and lubricants such as purified talc, stearin Salts, borax, polyethylene glycol and the like, as the corrigent sucrose, orange peel, citric acid, can be exemplified tartaric acid.

When an oral liquid preparation is prepared, a liquid preparation, a syrup, an elixir or the like can be produced by a conventional method by adding a corrigent, a buffer, a stabilizer, a corrigent and the like to the active ingredient. In this case, the flavoring agent may be those listed above, examples of the buffer include sodium citrate, and examples of the stabilizer include tragacanth, gum arabic, and gelatin.

When preparing injections, add pH regulators, buffers, stabilizers, tonicity agents, local anesthetics, etc. to active ingredients, and manufacture injections for subcutaneous, intramuscular and intravenous injections using conventional methods. can do. In this case, examples of the pH adjuster and buffer include sodium citrate, sodium acetate, and sodium phosphate. Examples of the stabilizer include sodium pyrosulfite, ethylenediaminetetraacetic acid (EDTA), thioglycolic acid, and thiolactic acid. Examples of local anesthetics include procaine hydrochloride and lidocaine hydrochloride. Examples of isotonic agents include sodium chloride and glucose.

When preparing a suppository, a formulation carrier known in the art, such as polyethylene glycol, lanolin, cocoa butter, fatty acid triglyceride, or the like, and a surfactant such as Tween (registered trademark) as necessary are used as active ingredients. After adding an agent etc., it can manufacture by a conventional method.

When preparing an ointment, bases, stabilizers, wetting agents, preservatives and the like that are usually used as active ingredients are blended as necessary, and mixed and formulated by a conventional method. Examples of the base include liquid paraffin, white petrolatum, white beeswax, octyldodecyl alcohol, and paraffin. Examples of the preservative include methyl paraoxybenzoate, ethyl paraoxybenzoate, propyl paraoxybenzoate, and the like.

When producing a patch, the ointment, cream, gel, paste and the like may be applied to a normal support by a conventional method. As the support, a woven fabric, nonwoven fabric, soft vinyl chloride, polyethylene, polyurethane film or foam sheet made of cotton, suf, chemical fiber is suitable.

The amount of the active ingredient contained in the drug can be appropriately determined depending on the dose of the active ingredient, the number of doses, and the like. The dose is not particularly limited, and the effectiveness of the contained components, administration form, administration route, type of disease, nature of the subject (such as weight, age, medical condition and use of other drugs), and judgment of the doctor in charge It is appropriately selected according to the above. The daily dose can be administered once to several times a day.

Human peripheral blood mononuclear cells (PBMC) were isolated from humans by Ficoll ^R gradient and cultured for 1 hour in RPMI medium containing 2 mM L-glutamine, Penicillin / Streptomycin, 10% inactivated fetal calf serum Each sample substance (piceatannol, passion fruit seed extract, isolapontigenin) (piceatannol, isolapontigenin obtained from Tokyo Kasei) is added at a predetermined concentration, further cultured for 1 hour, and then passion fruit seeds. In the experiment in which the extract was added, stimulation was performed with LPS (100 ng / ml), and in the experiment in which isolapontigenin was added, stimulation was performed with LPS (100 ng / ml) + IFNγ (20 U / ml = 1 ng / ml), followed by culture after 24 hours. The production amount of cytokine (IL-8, TNF-α) in the liquid was measured by ELISA. The results are shown in FIG. 1 (% inhibition is expressed as 100- ( cytokine production 24 hours after stimulation-control during non-stimulation) / (control during sample treatment- control during non-stimulation) x 100). The passion fruit seed extract was extracted as follows. Passion fruit seeds were roasted and pulverized, and 80% water-containing ethanol (80% (v / v) ethanol + 20% (v / v) water) was added thereto, mixed and stirred, and then 83 mesh filter (mesh size 180 μm) ) To separate solid and liquid. The obtained extract was concentrated by an evaporator and powdered by spray drying.

In addition, the aqueous ethanol extract of passion fruit seeds contained 9.49% (w / v) piceatannol, and the passion fruit seed extract corresponding to the added amount of purified piceatannol was used. .

As shown in FIG. 1, the passion fruit seed extract and isolapontigenin had an IL-8 production inhibitory activity and a TNF-α production inhibitory activity. Thus, passion fruit seed extract and isolapontigenin are remarkably effective as an anti-inflammatory agent, IL-8 production inhibitor and TNF-α production inhibitor.

Claims (4)

Anti-inflammatory agent containing passion fruit seed extract as an active ingredient (except for those caused by inflammation caused by reduced lymphatic function) .   An IL-8 production inhibitor comprising passion fruit seed extract as an active ingredient.   A TNF-α production inhibitor comprising passion fruit seed extract as an active ingredient.   Anti-inflammatory agent containing passion fruit seed extract as an active ingredient (excluding those for chronic inflammation caused by decreased lymphatic function).