WO2019146801A1 - Novel compound - Google Patents

Abstract

The present invention provides a compound represented by general formula (1) or a salt thereof. [In general formula (1), R¹ and R² represent an aldehyde group, a carboxyl group, or a C_1-6 alkyl group, and at least one of R1 and R2 is an aldehyde group or a carboxyl group. R³ represents a hydrogen atom, a C_1-6 alkyl group, a C_2-6 alkenyl group, a C_2-6 alkynyl group, a C_6-10 aryl group, a C_7-11 aralkyl group, a C_1-6 alkyl carbonyl group, or a C_7-11 aralkyl carbonyl group. R⁴ represents a hydrogen atom or a C_1-6 alkyl group.]

Description

New compound

The present invention relates to novel compounds.

Propolis is a wax-like composition obtained by solidifying resins and the like collected from various plants by honeybee insects, and has long been used as food and the like, and is known to have various physiological activities. For example, Patent Document 1 discloses a leukotriene production or release inhibitor containing propolis or an extract thereof as an active ingredient.

JP 2008-214235 A

Among the compounds derived from propolis, the present inventors have found novel compounds having physiological activities such as leukotriene release inhibition, NF-κB inhibition, and the like.

An object of the present invention is to provide a compound exhibiting leukotriene release inhibitory activity and NF--B inhibitory activity.

The present invention relates to a compound represented by the following general formula (1) or a salt thereof.

[In general formula (1),
R ¹ and R ^{2 each} represent an aldehyde group, a carboxyl group or a C _1-6 alkyl group, provided that at least one is an aldehyde group or a carboxyl group,
R ³ represents a hydrogen atom, a C _1-6 alkyl group, a C _2-6 alkenyl group, a C _2-6 alkynyl group, a C _6-10 aryl group, a C _7-11 aralkyl group, a C _1-6 alkylcarbonyl group, Or C _7-11 aralkylcarbonyl group,
R ⁴ represents a hydrogen atom or a C _1-6 alkyl group. ]

The compound represented by the above general formula (1) exhibits leukotriene release inhibitory action and NF-κB inhibitory activity.

In the general formula (1), R ¹ may be an aldehyde group or a carboxyl group, and R ² may be a C _1-6 alkyl group.

The present invention also relates to a compound represented by the formula (3) or a salt thereof.

The present invention also relates to a compound represented by the formula (4) or a salt thereof.

The present invention relates to a leukotriene release inhibitor containing, as an active ingredient, at least one selected from the group consisting of the aforementioned compounds and salts thereof.

The present invention also relates to an NF-κB inhibitor comprising, as an active ingredient, at least one selected from the group consisting of the aforementioned compounds and salts thereof.

According to the present invention, it is possible to provide a compound that exhibits leukotriene release inhibitory activity and NF-κB inhibitory activity.

It is a graph which shows the result of an Example.

Hereinafter, modes for carrying out the present invention will be described in detail. However, the present invention is not limited to the following embodiments. Various substituents defined or exemplified below can be arbitrarily selected and combined.

[The present compound]
One embodiment of the present invention is a compound represented by the general formula (1) (hereinafter, also referred to as "the present compound") or a salt thereof.

In the general formula (1), R ¹ and R ^{2 each} represent an aldehyde group, a carboxyl group or a C _1-6 alkyl group, provided that at least one is an aldehyde group or a carboxyl group,
R ³ represents a hydrogen atom, a C _1-6 alkyl group, a C _2-6 alkenyl group, a C _2-6 alkynyl group, a C _6-10 aryl group, a C _7-11 aralkyl group, a C _1-6 alkylcarbonyl group, Or C _7-11 aralkylcarbonyl group,
R ⁴ represents a hydrogen atom or a C _1-6 alkyl group.

In the present specification, “C _1-6 alkyl group” means a linear or branched alkyl group having 1 to 6 carbon atoms, and examples thereof include a methyl group, an ethyl group and an n-propyl group, Examples thereof include isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl and n-hexyl groups.

In the present specification, the “C _2-6 alkenyl group” means a linear or branched alkenyl group having 2 to 6 carbon atoms. As a C _2-6 alkenyl group, for example, vinyl group, propen-1-yl group, propen-2-yl group, 1-butenyl group, 2-butenyl group, 3-butenyl group, 1-methyl-1-propenyl group Group, 2-methyl-1-propenyl group, 1-pentenyl group, 2-pentenyl group, 3-pentenyl group, 4-pentenyl group, 5-pentenyl group, 1-methyl-1-butenyl group, 2-methyl-1 -Butenyl group, 3-methyl-1-butenyl group, 4-methyl-1-butenyl group, 1-methyl-2-butenyl group, 2-methyl-2-butenyl group, 3-methyl-2-butenyl group, 4 -Methyl-2-butenyl group, 1-methyl-3-butenyl group, 2-methyl-3-butenyl group, 3-methyl-3-butenyl group, 4-methyl-3-butenyl group, 1,2-dimethyl- 1-propenyl group, 1- And alkenyl groups such as hexenyl group, 2-hexenyl group, 3-hexenyl group, 4-hexenyl group, 5-hexenyl group, 6-hexenyl group and structural isomers thereof.

In the present specification, the “C _2-6 alkynyl group” means an alkynyl group having 2 to 6 carbon atoms. Examples of the C _2-6 alkynyl group include ethynyl group, propargyl group, butynyl group, pentynyl group and hexynyl group.

In the present specification, the “C _6-10 aryl group” means an aryl group having 6 to 10 carbon atoms. Examples of the C _6-10 aryl group include phenyl group and naphthyl group.

In the present specification, the “C _7-11 aralkyl group” means an alkyl group having a total carbon number of 7 to 11 and having an aryl group, and examples thereof include a benzyl group, a phenylethyl group and a naphthylmethyl group.

In the present specification, “C _1-6 alkylcarbonyl group” means a carbonyl group to which the above “C _1-6 alkyl group” is bonded, and examples thereof include an acetyl group, a propanoyl group, a pentanoyl group, a pivaloyl group and a heptanoyl group. And the like.

In the present specification, the “C _7-11 aralkylcarbonyl group” means a carbonyl group bonded to the above “C _7-11 aralkyl group”.

In General Formula (1), an aldehyde group, a carboxyl group or a C _1-6 alkyl group is shown, provided that at least one is an aldehyde group or a carboxyl group. One of R ¹ and R ² may be an aldehyde group or a carboxyl group, and the other may be a C _1-6 alkyl group. That is, the present compound may be a compound represented by the following general formula (2A), (2B), (2C) or (2D).

In formulas (2A), (2B), (2C) and (2D), R ³ and R ⁴ are as defined above.

The C _1-6 alkyl group in R ¹ and R ² is preferably a C _1-3 alkyl group which is an alkyl group having 1 to 3 carbon atoms, more preferably a methyl group.

In the general formula (1), preferably, R ¹ is an aldehyde group or a carboxyl group, R ² is a methyl group, more preferably, R ¹ is an aldehyde group, and R ² is a methyl group. The compound represented by the general formula (1) is preferably a compound represented by the above general formula (2A).

In the general formula (1), R ³ is a hydrogen atom, C _1-6 alkyl group, C _2-6 alkenyl group, C _2-6 alkynyl group, C _6-10 aryl group, C _7-11 aralkyl group, C _1-6 alkylcarbonyl group and C _7-11 aralkylcarbonyl group. R ³ is preferably a hydrogen atom.

In the general formula (1), R ⁴ is a hydrogen atom or a C _1-6 alkyl group. R ⁴ is preferably a hydrogen atom.

Preferably, at least one of R ³ and R ⁴ is a hydrogen atom, and more preferably both are a hydrogen atom.

As a specific example of a compound represented by General formula (1), the compound (Hereinafter, it is also mentioned "compound (3)") represented by following formula (3) is mentioned.

Moreover, the compound (Hereinafter, it is also mentioned "compound (4)") represented by following formula (4) is mentioned as another specific example of a compound represented by General formula (1).

The subject compounds may be salts acceptable for food or pharmaceutical use. Examples of salts of the present compound include salts with alkali metals, alkaline earth metals, other metals, ammonium and the like. More specific examples of the salt of the present compound include potassium salt, sodium salt, calcium salt and magnesium salt.

The present compound and its salt may be obtained by purification, for example, from natural products such as propolis. Specifically, for example, compound (3) can be obtained from a natural product by the method described in the examples described later. The present compound can also be obtained by chemical synthesis using a commercially available compound or an extract of a natural product (e.g., compound (3)) as a raw material. Specifically, for example, a functional group defined by R ³ and R ⁴ in the general formula (1) is introduced to the phenolic hydroxyl group and / or carboxyl group of the compound (3) by a known method. The extract may be obtained by introducing an aldehyde group by a known method to a natural product extract having no aldehyde group.

Among the compounds represented by the general formula (1), the compound (3) can be synthesized, for example, as follows. That is, first, the compound (C) is synthesized from the compound (A-1) or the compound (A-2) according to the following schemes 1 to 3. Subsequently, compound (3) can be obtained by deprotecting the protecting group of compound (C). The compound (A-1) or the compound (A-2) can be synthesized according to scheme 4 described later.

Scheme 1

In the formula, R ^a represents a methyl group (Me), R ^b represents a protecting group for a hydroxyl group such as an acyl group, an alkyl group or a silyl group, or a hydrogen atom, and R ^c represents an alkyl group etc. . In addition, a plurality of R ^a represents the same group. Specific examples of the silyl group include, for example, tert-butyldimethylsilyl group.

In the compound (A-1), one of the methyl groups (one of R ^a ) is oxidized with selenium dioxide to obtain a compound B (alcohol B) having a hydroxyl group. Subsequently, the compound (B) is oxidized to give a compound (C) having an aldehyde group. Mild conditions are desirable for oxidizing the compound (B) to obtain the compound (C), and in addition to metal oxidizing agents such as chromium and manganese, oxidizing agents such as organic sulfur reagent and halogen reagent are used. In addition, for example, compound (C) can also be obtained from compound (A-1) by oxidation with selenium dioxide.

Scheme 2

In the formula, R ^a represents a methyl group or a hydrogen atom, R ^b represents a protecting group for a hydroxyl group such as an acyl group, an alkyl group or a silyl group, or a hydrogen atom, and R ^c represents an alkyl group etc. . In addition, a plurality of R ^a represents the same group.

A compound (D) having an aldehyde group is obtained by selectively oxidatively cleaving the prenyl group of the compound (A-1) or the allyl group of the compound (A-2), 2- (triphenylphosphoranylidene) propion The compound (C) can be obtained by a carbon-addition reaction by Wittig reaction with an aldehyde.

Scheme 3

In the formula, R ^a represents a methyl group or a hydrogen atom, R ^b represents a protecting group for a hydroxyl group such as an acyl group, an alkyl group or a silyl group, or a hydrogen atom, and R ^c represents an alkyl group etc. . In addition, a plurality of R ^a represents the same group.

In addition, compound (C) can be synthesized by subjecting compound (A-1) or compound (A-2) to a cross metathesis reaction with methacrolein in the presence of a metal catalyst. As a catalyst used for metathesis reaction, Grubbs catalyst, Hoveyda-Grubbs catalyst, etc. are suitable.

The compound (A-1) or the compound (A-2) can be synthesized according to the following scheme 4.

Scheme 4

In the formula, X represents a halogen atom, R ^a represents a methyl group or a hydrogen atom, R ^b represents a protecting group for a hydroxyl group such as an acyl group, an alkyl group or a silyl group, or a hydrogen atom; ^c represents an alkyl group or the like. In addition, a plurality of R ^a represents the same group.

A commercially available 4-halophenol (compound (E)) is prenylated to give a compound (F-1). The halogen atom in the compound (E) is preferably a bromine atom or an iodine atom. At this time, a compound (F-2) which is an allyl derivative can be synthesized from 4-halophenol (compound (E)) using an allyl halide. The compound (F-1) or (F-2) is a compound (A-1) or a carbon chain extended by the Mizoroki-Heck reaction with an acrylic ester in the presence of a palladium catalyst, optionally after protecting the hydroxyl group. Let (A-2). The hydroxyl group in compound (F-1) or (F-2) may or may not be protected. When protecting a hydroxyl group, an acyl group, an alkyl group, a silyl group etc. are suitable for a hydroxyl-group protective group. Also, the ester portion of acrylic acid is preferably an ester with a fatty alcohol. Commercially available catalysts such as palladium acetate can be used as the palladium catalyst.

In the compound represented by the general formula (1), a compound in which one of R ^{1 and} R ² is a carboxyl group can be synthesized, for example, according to the following scheme. Specifically, compound (4) can be synthesized by, for example, Pinnick oxidation of compound (3) (Dorpanal).

Scheme 5

Compound (4) can also be synthesized from dolpanin (raw material) as shown in Scheme 6. Dorpanin (Drupanin) can be obtained by the method disclosed in Japanese Patent Application Laid-Open No. 2008-214235.

Scheme 6

In the formulae, P means a functional protecting group. Plural Ps in one molecule may be identical to or different from each other.

[Leukotriene Release Inhibitor]
The present compounds and salts thereof exhibit a leukotriene release inhibitory action, and thus are useful as leukotriene release inhibitors. That is, as an embodiment of the present invention, there is provided a leukotriene release inhibitor comprising, as an active ingredient, at least one selected from the group consisting of a compound represented by the above formula (1) and a salt thereof.

The leukotriene release inhibitor exhibits anti-allergic action, anti-inflammatory action, anti-pollinating action, anti-allergic rhinitis action, anti-atopic dermatitis action, anti-bronchial asthma action and the like through the leukotriene release inhibiting action. Therefore, the leukotriene release inhibitor can also be used as an anti-allergic agent, an anti-inflammatory agent, an anti-pollinating agent, an anti-allergic rhinitis agent, an anti-atopic dermatitis agent, an anti-bronchi asthma agent and the like.

The leukotriene release inhibitor of this embodiment may contain only the above-mentioned active ingredient, and may further contain other ingredients. As other ingredients, for example, pharmaceutically acceptable ingredients (eg, excipients, binders, lubricants, disintegrants, emulsifiers, surfactants, bases, solubilizers, suspending agents) Food-acceptable ingredients (eg, minerals, vitamins, flavonoids, quinones, polyphenols, amino acids, nucleic acids, essential fatty acids, refreshing agents, binders, sweeteners, disintegrants, lubricants, coloring agents Perfumes, stabilizers, preservatives, sustained release modifiers, surfactants, solubilizers, wetting agents).

The leukotriene release inhibitor of this embodiment can be used at a dose of 0.01 mg or more and 10 g or less per day for adults weighing 60 kg, preferably in a dose of 0.05 mg or more and 8 g or less in terms of active ingredient amount More preferably 0.10 mg to 3 g, still more preferably 0.15 mg to 1.5 g, still more preferably 0.20 mg to 1 g, It is even more preferable to use a dose of 22 mg to 500 mg, and it is particularly preferable to use a dose of 0.24 mg to 250 mg. The said dose can be suitably set within the said range according to factors, such as a person's health condition ingested, a method of administration, and a combination with other agents.

The leukotriene release inhibitor of this embodiment may be orally administered (ingested) or parenterally administered (eg, intranasal administration). The leukotriene release inhibitor of this embodiment may be administered once a day, or twice a day, three times a day, etc., as long as the amount of active ingredient per day is within the above-mentioned range. It may be administered separately. In addition, the leukotriene release inhibitor of the present embodiment is preferably administered continuously. By continuously administering, the leukotriene release inhibitory effect is more significantly exhibited.

The leukotriene release inhibitor of the present embodiment may be in any form of solid, liquid, paste and the like. The form of the leukotriene release inhibitor of the present embodiment may be, for example, uncoated tablets, sugar-coated tablets, granules, powders, tablets, capsules (hard capsules, soft capsules, seamless capsules). The leukotriene release inhibitor of this embodiment is formed into, for example, the above-mentioned dosage form by mixing at least one selected from the group consisting of the above-mentioned compounds as active ingredients and salts thereof and other components as necessary. It can be prepared by

The leukotriene release inhibitor of the present embodiment can be used as a drug, quasi drug and food composition itself as well as being added to a drug, quasi drug and food composition. The food composition is preferably a food in which the tertiary function (physical condition control function) of the food is emphasized. Examples of foods in which the tertiary functions of foods are emphasized include health foods, functional display foods, nutraceuticals, supplements, and foods for specific health.

A pharmaceutical comprising the leukotriene release inhibitor according to the present embodiment, a quasi-drug or food composition, or a pharmaceutical containing the leukotriene release inhibitor according to the present embodiment, the quasi-drug or food composition comprises leukotriene release suppression, It may be for anti-allergy, anti-pollinosis, anti-allergic rhinitis, anti-atopic dermatitis, anti-bronchial asthma. The above-mentioned product comprising a leukotriene release inhibitor or containing a leukotriene release inhibitor is, for example, to suppress allergies, to suppress anaphylactic symptoms, to suppress pollinosis, to suppress perennial rhinitis, itchy eyes or Congestion, suppression of discomfort, suppression of sneezing and cough, suppression of rhinitis, suppression of runny nose and congestion, suppression of atopic dermatitis, suppression of bronchial asthma, suppression of dyspnea, suppression of hives To suppress erythema and redness, to suppress swelling of the oral cavity, to suppress swelling of the eyelids, to suppress burning sensation, to suppress shortness of breath, to suppress nausea, to suppress discomfort of the digestive tract, vomiting and abdominal pain Indications of suppressing diarrhea and suppressing headache may be attached.

The content of the leukotriene release inhibitor of the present embodiment in the pharmaceuticals, quasi-drugs and food compositions is such that the amount of the active ingredient ingested per day is within the above-mentioned range, the pharmaceuticals, quasi-drugs and food compositions It may be set as appropriate according to the type of object and the like.

When the leukotriene release inhibitor of the present embodiment is used as a food composition itself or added to a food composition, the form of the food composition is not particularly limited. For example, beverages (coffee, juice, tea beverage, etc.) Soft drinks, milk drinks, lactic acid bacteria drinks, yogurt drinks, carbonated drinks etc .; spreads (custard cream etc.); pastes (fruit paste etc); Western confectionery (chocolate, donut, pie, shoe cream, gum, jelly, candy Cookies, cakes, puddings, etc .; Japanese confectionery (Daifuku, rice cakes, buns, castellas, antels, sheep, etc.); Frozen foods (ice cream, ice candy, sherbet etc.); , Soup, meat sauce, pasta, pickles, jam etc); seasonings (dressing, sprinkle, umami Seasonings, may be a arsenide) of soup.

The leukotriene release inhibitor of the present embodiment is used as a food in which the tertiary function of food is emphasized (for example, health food, functional display food, nutritional supplement, supplement or food for specified health use) itself, or the tertiary function of food In addition to the food forms mentioned above, for example, uncoated tablets, sugar-coated tablets, granules, powders, tablets, capsules, in addition to the food forms mentioned above, the food forms in which the tertiary function of the food is emphasized are used when added to the food emphasized. (Hard capsule, soft capsule, seamless capsule) may be used.

When the leukotriene release inhibitor according to the present embodiment is used as a drug or quasi drug itself or added to a drug or quasi drug, the form of the drug or quasi drug is not particularly limited. Tablets, dragees, granules, powders, tablets, capsules (hard capsules, soft capsules, seamless capsules) may be used.

The preparation method of the pharmaceutical, quasi-drug or food composition to which the leukotriene release inhibitor of this embodiment is added is not particularly limited, and any known method can be appropriately used. For example, a pharmaceutical, quasi-drug or food composition used for the above-mentioned application by mixing a leukotriene release inhibitor or the like into an intermediate product or final product in the manufacturing process of a drug, quasi-drug or food composition. You can get it.

[NF-κB inhibitor]
The compounds and salts thereof are useful as NF-κB inhibitors because they exhibit NF-κB inhibitory activity. That is, as one embodiment of the present invention, there is provided an NF-κB inhibitor comprising, as an active ingredient, at least one selected from the group consisting of the compound represented by the above formula (1) and a salt thereof.

The NF-κB inhibitor exhibits an anti-inflammatory action, an autoimmune disease ameliorating action, an arthritis ameliorating action, a rheumatoid arthritis ameliorating action, an osteoarthritis ameliorating action and the like through an NF-κB inhibitory action. Therefore, the NF-κB inhibitor can also be used as an anti-inflammatory agent, an autoimmune disease improving agent, an arthritis improving agent, a chronic rheumatoid arthritis improving agent, an osteoarthritis improving agent, and the like.

The NF-κB inhibitor of the present embodiment may contain only the above-mentioned active ingredient, and may further contain other ingredients. As other ingredients, for example, pharmaceutically acceptable ingredients (eg, excipients, binders, lubricants, disintegrants, emulsifiers, surfactants, bases, solubilizers, suspending agents) Food-acceptable ingredients (eg, minerals, vitamins, flavonoids, quinones, polyphenols, amino acids, nucleic acids, essential fatty acids, refreshing agents, binders, sweeteners, disintegrants, lubricants, coloring agents Perfumes, stabilizers, preservatives, sustained release modifiers, surfactants, solubilizers, wetting agents).

The NF-κB inhibitor of the present embodiment can be used at a dose of 0.01 mg or more and 10 g or less per day for adults weighing 60 kg, in a dose of 0.05 mg or more and 8 g or less in terms of active ingredient amount It is preferable to use a dose of 0.10 mg or more and 3 g or less, more preferably a dose of 0.15 mg or more and 1.5 g or less, and still more preferably a dose of 0.20 mg or more and 1 g or less, It is even more preferred to use doses of 0.22 mg to 500 mg, and particularly preferred to use doses of 0.24 mg to 250 mg. The said dose can be suitably set within the said range according to factors, such as a person's health condition ingested, a method of administration, and a combination with other agents.

The NF-κB inhibitor of the present embodiment may be orally administered (ingested) or parenterally administered (eg, intranasal administration). The NF-κB inhibitor of the present embodiment may be administered once a day, as long as the amount of active ingredient per day is within the above-mentioned range, or twice or three times a day, etc. multiple times May be divided and administered. In addition, the NF-κB inhibitor of the present embodiment is preferably administered continuously. By continuously administering, the NF-κB inhibitory effect is more significantly exerted.

The NF-κB inhibitor of the present embodiment may be in any form of solid, liquid, paste and the like. The form of the NF-κB inhibitor of the present embodiment may be, for example, uncoated tablets, sugar-coated tablets, granules, powders, tablets, capsules (hard capsules, soft capsules, seamless capsules). The NF-κB inhibitor of the present embodiment is, for example, a mixture of at least one selected from the group consisting of the above-mentioned active ingredient and the salt thereof and other components as necessary, in the above dosage form It can be prepared by molding.

The NF-κB inhibitor of this embodiment can be used as a medicine, quasi-drug and food composition itself as well as being added to a medicine, quasi-drug and food composition. The food composition is preferably a food in which the tertiary function (physical condition control function) of the food is emphasized. Examples of foods in which the tertiary functions of foods are emphasized include health foods, functional display foods, nutraceuticals, supplements, and foods for specific health.

A pharmaceutical, quasi-drug or food composition comprising the NF-κB inhibitor of the present embodiment, or a pharmaceutical, quasi-drug or food composition containing the NF-κ B inhibitor of the present embodiment is NF-κB inhibition For anti-inflammatory, for improving autoimmune diseases, for improving arthritis, for improving rheumatoid arthritis, for improving osteoarthritis, and for anti-cancer. The above-mentioned product consisting of an NF-κB inhibitor or comprising an NF-κB inhibitor has, for example, an effect of suppressing inflammation, an effect of modulating immune function, an effect of preventing an autoimmune disease, and an inhibition of a viral disease Effect to relieve joint pain, to prevent rheumatoid arthritis, to prevent deterioration of atopic dermatitis, to prevent Crohn's disease and inflammatory bowel disease, to suppress the onset and progression of tumor, to cause cancer Purportedly suppress proliferation, progression (proliferation, invasion, metastasis), prevent septicemia, suppress proliferation of cytomegalovirus (CMV) or human immunodeficiency virus (HIV), prevent metabolic syndrome, To prevent locomotive syndrome, to suppress muscle atrophy, to prevent sarcopenia, to suppress walking decline, to suppress disc herniation, to suppress back pain, to prevent osteoporosis In addition, indications to suppress bone aging, to suppress epidermal thickening, to suppress pigmentation, to suppress a decrease in skin elasticity, and to suppress photoaging of a skin may be attached.

The content of the NF-κB inhibitor of the present embodiment in the pharmaceuticals, quasi-drugs and food compositions is such that the amount of active ingredient to be ingested per day is within the above-mentioned range. It may be appropriately set according to the type of composition and the like.

When the NF-κB inhibitor of the present embodiment is used as a food composition itself or added to a food composition, the form of the food composition is not particularly limited. For example, those exemplified for the above-mentioned leukotriene release inhibitor And may be similar to

The NF-κB inhibitor of the present embodiment may be used as a food in which the tertiary function of food is emphasized (for example, health food, functional display food, nutritional supplement, supplement or food for specific health use) itself or as a food tertiary In addition to the above-described food forms, for example, uncoated tablets, sugar-coated tablets, granules, powders, tablets, in addition to the food forms mentioned above, food forms in which the tertiary function of food is emphasized are used when added to food in which function is emphasized. It may be a capsule (hard capsule, soft capsule, seamless capsule).

When the NF-κB inhibitor of the present embodiment is used as a drug or quasi drug itself or added to a drug or quasi drug, the form of the drug or quasi drug is not particularly limited. Uncoated tablets, dragees, granules, powders, tablets, capsules (hard capsules, soft capsules, seamless capsules) may be used.

The preparation method of the pharmaceutical, quasi-drug or food composition to which the NF-κB inhibitor of the present embodiment is added is not particularly limited, and any known method can be appropriately used. For example, pharmaceuticals, quasi-drugs or food compositions used for the above-mentioned applications by mixing an NF-κB inhibitor or the like with an intermediate product or final product in the manufacturing process of pharmaceuticals, quasi-drugs or food compositions You can get

Hereinafter, the present invention will be more specifically described based on examples. However, the present invention is not limited to the following examples.

[Test Example 1: Purification and Structure Determination of Test Compound]
[Purification of test compound]
71 g of an 80% ethanol extract (Lot. 451A) of an alexin propolis bulk (manufactured by Minas Gerais) was dissolved in 560 mL of ethyl acetate, and transferred to a separatory funnel. 700 mL of ultrapure water was added, and 0.1 N hydrochloric acid was added while confirming with pH paper to adjust the pH to 2. Liquid-liquid extraction was performed, and the ethyl acetate layer was recovered. The aqueous layer was extracted twice more with the same amount of ethyl acetate. The ethyl acetate layer was concentrated by an evaporator to obtain 38.6 g of ethyl acetate extract.

Column chromatography (Type of gel: Silica gel made by Osaka Soda Co., Ltd. (IR-60-40 / 63, particle diameter 50 μm, pore diameter 6 nm, surface area 450 m ² / g, pore volume 0.70 mL / column) g, water content 2.0% or less), gel amount: 1000 g, column size: φ 80 × 630 mm, elution conditions: 200 mL of 100% hexane, 10 L of 50% chloroform-hexane, 8 L of 70% chloroform-hexane, 100% 5 L of chloroform, 5 L of 10% ethyl acetate-chloroform, 5 L of 20% ethyl acetate-chloroform, and 10 L of methanol were flowed and fractionated into 13 fractions as follows.
(1) 100% hexane 200 mL fraction (9.8 mg)
(2) 50% hexane-chloroform 0 to 10 L fraction (883.3 mg)
(3) 70% hexane-chloroform 0 to 7 L fraction (1967.6 mg)
(4) 70% hexane-chloroform 7-8th fraction (6248.7 mg)
(5) 100% chloroform 0-1 L-th fraction (6480 mg)
(6) 100% chloroform 1-2 L eye fraction (631.6 mg)
(7) 100% chloroform 2-3 L fraction (123. 78 mg)
(8) 100% chloroform 3-5 L fraction (2007.1 mg)
(9) 10% ethyl acetate-chloroform 0-2 L fraction (643.1 mg)
(10) 10% ethyl acetate-chloroform 2-5 L-th fraction (3885.9 mg)
(11) 20% ethyl acetate-chloroform 0-3 L-th fraction (2197.4 mg)
(12) 20% ethyl acetate-chloroform 3-5 L eyes fraction (1082.58 mg)
(13) Methanol fraction (13823.58 mg)

(12) 20% ethyl acetate-chloroform 3-5 L fraction (1082.58 mg) was fractionated by medium pressure column chromatography under the following conditions.
(Medium pressure column chromatography conditions)
Column used: ODS-SM 50 μm (gel amount: 110 g, column size: φ 46 × 130 mm)
Flow rate: 60 mL / min
Fractional volume: 100 mL
Elution conditions: 40%, 50%, 60%, 70% methanol-0.1% TFA, 20 minutes each

The fractions obtained by medium pressure column chromatography are as follows.
-40% MeOH fraction: fraction number (Fr.) 1-12
-50% MeOH fraction: Fr. 1-12
60% MeOH fraction: Fr. 1-12
-70% MeOH fraction: Fr. 1-12

50% MeOH fraction Fr. 3-7 (54.31 mg) was separated by HPLC (preparation conditions: Cosmosil 5C18-AR-II (10 mm × 250 mm), 40% methanol-0.1% TFA).

The sample obtained by HPLC fractionation was dissolved in ethyl acetate and then recrystallized by adding hexane to obtain a compound of unknown structure.

[Determination of structure of test compound]
The obtained compound was analyzed by the following apparatus to determine the structure.
Mass spectrometer: ACQUITY UPLC-Quattro Premier XE manufactured by Waters
NMR: Bruker Biospin AVANCE 500

LC-MS measurement conditions (UPLC conditions)
Equipment: Waters Acquity
Column: Sunniest C18-HT, 2 μm, 2.1 x 100 mm
Column oven: 40 ° C
Amount of implantation: 2 μL
Flow rate: 0.3 mL / min
Solvent: A = 0.1% aqueous formic acid, B = methanol Elution conditions: B 5% (2 min hold), 5-40% (8 min linear gradient), 40-85% (20 min linear gradient), 100% (4 min hold), 100-5% (2 min linear gradient), 5% (4 min hold) 40 minutes in total (MS / MS conditions)
Equipment: Quattro Premier Mass Spectrometer (Waters)

As a result of MS measurement, the molecular weight of the structural unknown compound was estimated to be 246.

NMR measurements were performed on structurally unknown compounds. Table 1 shows the ¹ H-NMR and ¹³ C-NMR measurement data.

The obtained structural unknown compound is obtained from (2E) -3- {4-hydroxy-3-[(2E) -3-methyl-4-oxobut-2- nyl] phenyl} -2-2 based on the measurement results of NMR and MS. It was decided as propenoic acid and named as dolpanal.

[Structural determination of synthetic compounds]
(2E) -3- {4-hydroxy-3-[(2E) -3-methyl-4-oxobut-2-yl] phenyl} -2-propenoic acid (Dolpanal) described in Synthesis Example 1 below Synthesized according to The ¹ H-NMR, ¹³ C-NMR and MS measurements were carried out on the synthesized compounds. The measuring apparatus used the same apparatus as the apparatus used for the structure determination of dolpanar.

As a result of MS measurement, the molecular weight of the synthesized compound was estimated to be 246. Table 2 shows ¹ H-NMR and ¹³ C-NMR measurement data.

[Test Example 2: Leukotriene Release Inhibitory Activity]
[Preparation of materials]
First, the following materials were prepared.
(1) HL-60 cells HL-60 cells were cultured in RPMI 1640 medium containing 10% FBS, 100 units / ml penicillin, 100 μg / ml streptomycin (37 ° C., 5% CO ₂ ).
(2) Leukotriene assay reagent Product name: CAST (registered trademark) (cellular antigen stimulation test)-2000 ELISA (supplier: BUHLMANN)
(3) Other reagents Calcium ionophore A23187 (SIGMA C 7522), BSA (SIGMA A8806), DMSO (Nacalai GR13407-45), D-PBS (-) (Nacalai Tesque 14249-95), D-PBS (+) (Nacalai Tesque 14248-05), nitro blue tetrazolium (NBT) (Nacalai Tesque 24720-56) and Phorborl 12-myristate 13-acetate (PMA) (SIGMA P8139) were purchased from commercial sources and used.

[Sample preparation]
Dolpanal was dissolved in DMSO to a concentration of 3 mg / ml. After dilution with PBS (+), it was added to the test system according to the target test concentration. The final concentration of DMSO was adjusted to 1% (v / v).

[Leukotriene measurement]
(Differentiation of HL-60 cells)
HL-60 cells were cultured in RPMI 1640 (10% FBS, 100 units / ml penicillin, 100 μg / ml streptomycin) medium and used for the test (37 ° C., 5% CO ₂ ). After adjusting the cells to 5 × 10 ⁵ cells / ml, 1.25% DMSO was added and differentiation was induced by incubating at 37 ° C., 5% CO ₂ for 6 days. To determine differentiation to granulocyte-like cells, add 1 mg / ml NBT and 4 μMPMA equally to the cell suspension, incubate at 37 ° C for 30 minutes, and calculate the percentage of positive cells under a microscope (NBT reduction method).

[Leukotriene production induction]
Each sample was added to DMSO-differentiated HL-60 cells (suspended in PBS (+)-1% BSA) and preincubated at 37 ° C for 15 minutes, then 1 μM A23187 was added and incubated at 37 ° C for 15 minutes (Cell concentration 1 × 10 ⁶ cells / ml, containing DMSO 1% to 2%). The supernatant collected was used as a measurement sample after inducing the production of leukotriene.

[CysLT amount measurement]
The leukotriene concentration in the recovered supernatant was quantified by CAST ELISA kit (n = 2 for each condition).

[Calculation of IC ₅₀ ]
The log of the test concentration of each sample is plotted on the horizontal axis, and the leukotriene release inhibition rate is plotted on the vertical axis. IC ₅₀ was calculated based on the following formula using regression calculation software (Kaiki 6) from the concentration at two points surrounding 50% and the inhibition rate (%).
Calculation formula: IC ₅₀ = 10 ^{(log [A / B] * [50-C] / [DC] + log [B])}
A = 50% high concentration B = 50% low concentration C = B inhibition ratio inhibition ratio D = A inhibition ratio

〔Test results〕
The IC ₅₀ of leukotriene release inhibitory activity of dolpanal was 0.90 μg / mL. On the other hand, the IC ₅₀ of leukotriene release inhibitory activity of altepilin C and dulpannin which are cinnamic acid derivatives disclosed in JP 2008-214235 A was 3.0 μg / mL and 7.0 μg / mL, respectively.

Brazilian green propolis _{IC 50} of itself containing Dorupanaru was 1.46μg / mL.

[Test Example 3: NF-κB Inhibitory Activity]
〔material〕
The following materials were prepared.
Cell line: NF-κ Reporter, Luciferase, HEK293 Recombinant Cell Line (BPS Bioscience)
Media: Dulbecco's Modified Eagle's Medium-low glucose-(Thermo Fisher Scientific)
FBS: Serum, Fetal Bovine, BSE Tested, EC Approved (biowest)
Measurement plate: ViewPlate-384 TC (PerkinElmer)

〔Measuring method〕
The cultured cell lines (NF-κ Reporter, Luciferase, HEK293 Recombinant Cell Line) were seeded at a concentration of 10,000 cells / well on two 384-well plates for cell culture. The seeded cells were incubated at 37 ° C. overnight.

Dolpanal was adjusted to a final concentration of 50, 10, 2, 0.4 μg / mL, 1% DMSO with dimethyl sulfoxide (DMSO), and added to cells of two plates. Spin down was performed under the conditions of 220 × g and 10 seconds. Then, it incubated at 37 degreeC for 1 hour.

TNF-α (R & D Systems), adjusted to a final concentration of 10 ng / mL with DMEM medium, was added to the cells of two plates. Spin down was performed under the conditions of 220 × g and 10 seconds. Then, it incubated at 37 degreeC for 5 hours. Of the two identical plates, one was subjected to cell viability measurement and the other to luciferase assay. CellTiter-Glo (registered trademark) Luminescent Cell Viability Assay (Promega) was used for cell survival number measurement.

First, the medium and an equal volume of CellTiter-Glo reagent were added to the cells, and shaken for 2 minutes while shielding light. Then, it incubated at room temperature for 10 minutes, and measured the fluorescence intensity in EnVision (PerkinElmer). Luciferase assay using ^{ONE-Glo TM Luciferase Assay System (} Promega). The medium and an equal amount of ONE-Glo reagent were added to the cells, and shaken for 1 minute while blocking light. After incubation for 3 minutes at room temperature, the fluorescence intensity was measured with EnVision (PerkinElmer).

The value of the luciferase assay result is divided by the number of cell survival. Furthermore, the value was normalized to the value where the fluorescence intensity of the well to which only TNF-α and the solvent were added was 100% and the well to which no TNF-α was added was 0%. From the results obtained, 50% inhibitory concentration was calculated using statistical software GraphPad Prism 7.

〔Test results〕
FIG. 1 shows the results of NF-κB inhibitory activity of dolpanal. As a result, _{IC 50} of the NF-[kappa] B inhibitory activity of Dorupanaru was found to 13.4μg / mL. As a positive control, the _{IC 50} of Ro106-9920 commonly used is the average 5.3μM (= 1.3μg / mL) (measured value).

Synthesis Example 1: Chemical Synthesis 1 of Dolpanal
Chemical synthesis of dolpanal was carried out according to Scheme I below.

4-iodophenol (compound 2), sodium hydride (NaH), and 1-bromo-3-methyl-2-butene were mixed in toluene at 0 ° C. and then stirred at room temperature (rt). The reaction mixture was post-treated in a conventional manner to synthesize Compound 3 in a yield of 62%.

The compound 3, t-butyl acrylate, palladium (II) acetate, tri (o-tolyl) phosphine, tetrabutyl ammonium chloride, triethylamine and water obtained by the above method are reacted in dimethylformamide (DMF) at 35 ° C. Compound 4 was synthesized in 96% yield by post-treating the resulting reaction mixture according to a conventional method.

Compound 4 is reacted with selenium dioxide in an aqueous solution of 95% ethanol at 60 to 65 ° C., and the resulting reaction mixture is worked up by a conventional method to give compound 5 (yield 43%) and compound 6 (30%). %) Was synthesized. The yield of compound 5 and compound 6 is a yield based on the consumption of compound 4 which is a starting material.

Compound 1 was synthesized in a yield of 94% by reacting compound 6 in dichloromethane and water in the presence of trifluoroacetic acid, and post-treating the resulting reaction mixture according to a conventional method.

Synthesis Example 2: Chemical Synthesis of Dolpanal 2
Chemical synthesis of dolpanal was carried out according to Scheme II below.

Compound 4 was synthesized from compound 2 in the same manner as in Scheme I.

Compound 4, tert-butyldimethylsilyl chloride (TBSCl), and imidazole were added in dimethylformamide (DMF) at 0 ° C. and stirred at room temperature. Compound 7 was synthesized in a yield of 85% by post-treating the resulting reaction mixture according to a conventional method.

Compound 7 is reacted with selenium dioxide in an aqueous solution of 95% ethanol at 60 to 65 ° C., and the resulting reaction mixture is post-treated by a conventional method to give compound 8 (yield 42%) and compound 9 (22) %) Was synthesized. Compound 8 was reacted in dichloromethane in the presence of manganese dioxide, and the resulting mixture was worked up according to a conventional method to give compound 9 (yield 63%).

The obtained compound 9, tert-butyldimethylsilyl triflate and 2,6-lutidine were added to dichloromethane at 0 ° C. and reacted at room temperature. The reaction mixture obtained was worked up and then reacted in dichloromethane in the presence of 1 mol / l hydrochloric acid. The reaction mixture thus obtained is worked up in the usual manner and reacted in the presence of tetrabutylammonium fluoride in tetrahydrofuran at room temperature. The reaction mixture thus obtained was worked up by a conventional method to synthesize Compound 1.

Compound 2 synthesized according to Scheme II is (2E) -3- {4-hydroxy-3-[(2E) -3-methyl-4-oxobut-2-yl] phenyl}-isolated from a bulk mass of alexin propolis It was confirmed that the NMR and MS measurement data were in agreement with 2-propenoic acid (Dolpanal) and Compound 1 synthesized according to Scheme I.

Claims (6)

1. The compound or its salt represented by following General formula (1).
   

   

   [In general formula (1),
   R ¹ and R ^{2 each} represent an aldehyde group, a carboxyl group or a C _1-6 alkyl group, provided that at least one is an aldehyde group or a carboxyl group,
   R ³ represents a hydrogen atom, a C _1-6 alkyl group, a C _2-6 alkenyl group, a C _2-6 alkynyl group, a C _6-10 aryl group, a C _7-11 aralkyl group, a C _1-6 alkylcarbonyl group, Or C _7-11 aralkylcarbonyl group,
   R ⁴ represents a hydrogen atom or a C _1-6 alkyl group. ]
2. Wherein R ¹ is an aldehyde group or a carboxyl group, wherein R ² is a C _1-6 alkyl group, a compound or a salt thereof according to claim 1.
3. The compound or its salt represented by following formula (3).
   

   
4. The compound or its salt represented by following formula (4).
   

   
5. A leukotriene release inhibitor comprising, as an active ingredient, at least one selected from the group consisting of a compound according to any one of claims 1 to 4 and a salt thereof.
6. An NF-κB inhibitor comprising, as an active ingredient, at least one selected from the group consisting of a compound according to any one of claims 1 to 4 and a salt thereof.

Images