JP6188422B2 - Anti-inflammatory agent, hyaluronidase inhibitor, antiallergic agent - Google Patents

Description

    The present invention relates to an anti-inflammatory agent, a hyaluronidase inhibitor, and an antiallergic agent having a high hyaluronidase inhibitory action.

  Hyaluronic acid is a kind of mucopolysaccharide that is abundant in tissues such as skin and joint fluid.For example, in the skin, cell protection, nutrient transport, tissue moisture retention, flexibility maintenance, etc. As synovial fluid, it plays an important role in maintaining the tissue structure / function and maintaining lubricity.

  The amount of hyaluronic acid in living bodies such as skin and joints decreases due to aging or pathological conditions, which may cause dry skin, decreased elasticity, increased wrinkles, rough skin, joint pain due to deterioration of joint wettability, etc. Raise. On the other hand, by inhibiting hyaluronidase, which is an enzyme that hydrolyzes hyaluronic acid, it is possible to suppress the degradation of hyaluronic acid and contribute to the maintenance of the amount of biological hyaluronic acid.

  It is known that hyaluronidase is activated at the time of inflammation, thereby destroying the matrix of connective tissue and enhancing the infiltration of inflammatory tissues and the permeability of blood vessels. It is also known that hyaluronidase mediates the process of histamine release from mast cells in type I allergy. Therefore, the hyaluronidase inhibitor can be used as an anti-inflammatory / anti-allergic agent caused by hyaluronidase.

  Therefore, hyaluronidase inhibitors are increasing in number of patients in recent years, and are expected to prevent or improve allergic diseases such as atopic dermatitis, hay fever, bronchial asthma and food allergies, which are becoming social problems. For example, anti-allergic sodium cromoglycate and tranilast, and anti-inflammatory drugs aspirin and indomethacin have hyaluronidase activity inhibitory activity, suggesting that this inhibitory role plays a role in exerting therapeutic effects. (Non-patent Documents 1 and 2).

  Conventionally, as a hyaluronidase inhibitor, carrageenan, alginic acid, hydrolyzate of alginic acid (see Patent Document 1), manly extract (see Patent Document 2), low molecular weight psyllium seed gum (see Patent Document 3), and the like are known. However, the effect was not sufficient.

  As a derivative of chlorogenic acid, it is known that ethyl ester of chlorogenic acid exerts an anti-influenza virus effect (see Patent Document 4).

JP 2013-10700 A JP2013-6815A JP2012-193134A JP 2004-345971 A

Chem. Pharm. Bull. 33, 642 (1985) Kakegawa Toshio et al., Inflammation, 4,437 (1984)

  The present invention provides an anti-inflammatory agent, a hyaluronidase inhibitor, and an antiallergic agent having a high hyaluronidase inhibitory action.

  The present invention relates to a compound represented by the following chemical formula (1)

(In formula 1, R ¹ represents H or a saturated or unsaturated alkyl group having a linear or branched chain having 1 to 12 carbon atoms, and R ^{2 to} R ⁶ may be the same or different, and H or carbon number. An anti-inflammatory agent, a hyaluronidase inhibitor, an anti-inflammatory agent containing 2 to 8 straight-chain or branched-chain, saturated or unsaturated acyl groups, and R ^{1 to} R ⁶ are not all H) It is an allergic agent.

In the second invention of the present application, in the chemical formula (1), R ¹ is one or two selected from an ethyl group and a 2-ethylhexyl group, and a compound in which R ^{2 to} R ⁶ are H is an active ingredient. It is an anti-inflammatory agent.

The third aspect of the invention, in the chemical formula (1), ^{R 1} is ^H, is 1 or more acetyl groups R 2 to R ^6, the active ingredient a compound ^R 2 to R ⁶ other than acetyl group is H The anti-inflammatory agent according to claim 1.

Fourth aspect of the invention, in the chemical formula (1), R ¹ is an anti-inflammatory agent according to claim 1 H, all R ² to R ⁶ is the compound as an active ingredient is an acetyl group.

  5th invention of this application is a hyaluronidase inhibitor which uses the compound of 1 of Claims 1-4 as an active ingredient.

  6th invention of this application is an antiallergic agent which uses the compound of Claim 1-4 as an active ingredient.

  Since the compound described in the chemical formula (1) of the present invention has a high hyaluronidase inhibitory effect, it exhibits an antiallergic effect and an antiinflammatory effect.

  Hereinafter, modes for carrying out the present invention will be described.

  The present invention relates to a compound represented by the following chemical formula (1)

(In formula 1, R ¹ represents H or a saturated or unsaturated alkyl group having a linear or branched chain having 1 to 12 carbon atoms, and R ^{2 to} R ⁶ may be the same or different, and H or carbon number. An anti-inflammatory agent, a hyaluronidase inhibitor, an anti-inflammatory agent containing 2 to 8 straight-chain or branched-chain, saturated or unsaturated acyl groups, and R ^{1 to} R ⁶ are not all H) It is an allergic agent.

From the viewpoint of the effect of the present invention, in the chemical formula (1), a compound in which R ¹ is one or two selected from an ethyl group and a 2-ethylhexyl group, and R ^{2 to} R ⁶ are H is preferably used. .

From the viewpoint of the effect of the present invention, in the chemical formula (1), ^{R 1} is ^H, is 1 or more acetyl groups R 2 to R ^6, the compound ^R 2 to R ⁶ other than acetyl groups are H are preferably used It is done.

From the viewpoint of the effect of the present invention, in the chemical formula (1), a compound in which R ¹ is H and all of R ^{2 to} R ⁶ are acetyl groups is preferably used.

The compound of the present invention can be synthesized using chlorogenic acid, which is a compound represented by the following chemical formula (2), as a starting material.

When an alkyl group is bonded to R ¹ by ester bonding, the compound of formula (2) and an alcohol having 1 to 12 carbon atoms are mixed in a dilute aqueous acid solution and reacted at room temperature or under a temperature condition of 60 ° C. or less. Can be synthesized.

When an acyl group is bonded to R ^{2 to} R ⁶ , synthesis is performed by mixing the compound of the chemical formula (2) and a carboxylic anhydride-pyrimidine mixed solution and reacting them at room temperature or at a temperature of 60 ° C. or less in a timely manner. Can do. At this time, for the hydroxyl group to which no acyl group is bonded, the masking can be removed after the reaction after masking with another compound in advance.

  The compound represented by the chemical formula (1) has a high hyaluronidase inhibitory effect and is useful as an anti-inflammatory agent, a hyaluronidase inhibitor, and an antiallergic agent.

  The anti-inflammatory agent, hyaluronidase inhibitor, and antiallergic agent of the present invention may be composed only of the compound represented by the chemical formula (1) or may be formulated. The type of allergy to be targeted is not limited, and examples include atopic dermatitis, hay fever, bronchial asthma, food allergy, and the like.

  The anti-inflammatory agent, hyaluronidase inhibitor, and antiallergic agent of the present invention can be used as pharmaceuticals (including pharmaceuticals and quasi drugs). The above pharmaceutical products can be used for humans or for animals other than humans.

  The anti-inflammatory agent, hyaluronidase inhibitor, and antiallergic agent of the present invention can be appropriately selected depending on the dosage form. It can be produced by appropriately blending. Examples of additives that can be incorporated into the anti-inflammatory agent, hyaluronidase inhibitor, and antiallergic agent of the present invention include excipients (glucose, lactose, sucrose, sodium chloride, starch, calcium carbonate, kaolin, crystalline cellulose, cocoa butter, etc. , Hydrogenated vegetable oil, kaolin, talc, etc.), binder (distilled water, physiological saline, ethanol water, simple syrup, glucose solution, starch solution, gelatin solution, carboxymethylcellulose, potassium phosphate, polyvinylpyrrolidone, etc.), disintegrant ( Kantene, sodium bicarbonate, calcium carbonate, sodium lauryl sulfate, stearic acid monoglyceride, starch, lactose, gelatin, ethanol, etc.), disintegration inhibitors (sucrose, stearin, cocoa butter, hydrogenated oil, etc.), absorption enhancers (fourth) Secondary ammonium base, sodium lauryl sulfate, etc.) Agents (glycerin, starch, lactose, kaolin, bentonite, silicic acid, etc.), lubricants (purified talc, stearates, polyethylene glycol, and the like) and the like.

  The administration method of the anti-inflammatory agent, hyaluronidase inhibitor and antiallergic agent according to the present invention is generally administered orally in the form of tablets, pills, soft / hard capsules, fine granules, powders, granules, etc. be able to. The water-soluble preparation can be administered orally as a liquid. Furthermore, parenteral administration may be used. When administered as a parenteral agent, the anti-inflammatory agent, hyaluronidase inhibitor and antiallergic agent of the present invention are dispersed in a suitable solubilizing agent such as ethanol or water, and then a poultice, lotion, ointment, tincture It can be applied in a dosage form such as a cream. The water-soluble preparations of this anti-inflammatory agent, hyaluronidase inhibitor, and antiallergic agent are used as they are or with the addition of a dispersing agent, suspending agent, stabilizing agent, etc. It can be applied in a dosage form such as a cream.

  The compounding ratio when using the anti-inflammatory agent, hyaluronidase inhibitor and antiallergic agent of the present invention as a drug can be appropriately changed depending on the dosage form, but is usually administered orally or by mucosal absorption. In the case of parenteral administration, about 0.01 to 10 wt% is recommended. In addition, since the dose varies depending on various conditions, a dose smaller than the above dose may be sufficient, or it may be necessary to administer beyond the range. The pharmaceutical composition includes, in addition to the anti-inflammatory agent, hyaluronidase inhibitor, and antiallergic agent, other known compounds commonly used in the pharmaceutical field, and compounds necessary for molding into a form suitable for oral administration. It may be. Examples of such compounds include lactose, starch, hydroxypropyl cellulose, kaolin, talc, calcium carbonate and the like.

  The anti-inflammatory agent, hyaluronidase inhibitor and anti-allergic agent of the present invention are expected to have anti-inflammatory action, hyaluronidase inhibitory action and anti-allergic action even when used as an external preparation for skin (including cosmetics, pharmaceuticals and quasi drugs). can do. In addition, the said skin external preparation may be used for a human and may be used for mammals other than a human. Examples of the external preparation for skin to which the hyaluronidase inhibitor, antiallergic agent and anti-inflammatory agent of the present invention can be incorporated include, for example, emulsion, soap, facial cleanser, bath preparation, cream, emulsion, lotion, eau de cologne, shaving Cream, shave lotion, cosmetic oil, suntan / sunscreen lotion, funny powder, foundation, perfume, pack, nail cream, enamel, enamel remover, eyebrow, blusher, eye cream, eye shadow, mascara, eyeliner, lipstick Lip balm, shampoo, rinse, hair dye, dispersion, detergent and the like. Examples of the form of pharmaceuticals or quasi drugs that can contain the anti-inflammatory agent, hyaluronidase inhibitor, and antiallergic agent of the present invention include ointments, creams, and liquids for external use.

  In addition to the anti-inflammatory agent, hyaluronidase inhibitor, and antiallergic agent according to the present invention, the external preparation for skin of the above form includes cosmetics and quasi-drugs as long as the anti-inflammatory action, hyaluronidase inhibitory action, and antiallergic action are not impaired. Ingredients blended in skin external preparations such as oils, higher alcohols, fatty acids, UV absorbers, powders, pigments, surfactants, polyhydric alcohols / sugars, polymers, bioactive ingredients, solvents, antioxidants, A fragrance | flavor, antiseptic | preservative, etc. can be mix | blended.

  Since the compound represented by the chemical formula (1) of the present invention can be taken orally, it can also be used as a food or drink. In this case, the composition of the present invention can be eaten as it is or contained in an enteric capsule for administration. Or it can melt | dissolve in liquid (for example, water) so that it may become a suitable density | concentration, and it can add to food etc. by methods, such as mixing, immersion, application | coating, and spraying.

  In the food and drink of the present invention, the content ratio of the compound represented by the chemical formula (1) is not limited, and can be set as appropriate according to various uses. Moreover, also when obtaining the food / beverage products of this invention, in the well-known manufacturing method according to various uses, the compound shown to Chemical formula (1) can be contained by arbitrary methods or timing. The form of the compound represented by the chemical formula (1) at the time of inclusion is not limited, and may be a slurry or a powder, and may be appropriately selected according to various uses.

  The food / beverage products of the present invention are not particularly limited, and can include all foods that humans can eat. For example, edible oil (salad oil), confectionery (gum, candy, caramel, chocolate, cookie, snack, jelly, gummi, tablet confection etc.), noodles (soba, udon, ramen etc.), dairy products (milk, ice cream, Yogurt, etc.), seasonings (miso, soy sauce, etc.), soups, beverages (juice, coffee, tea, tea, carbonated beverages, sports drinks, etc.) and general foods, health foods (tablets, capsules, etc.), Nutritional supplements (such as energy drinks) are listed. A compound represented by the chemical formula (1) of the present invention may be appropriately blended with these foods and drinks.

  The addition amount of the composition of this invention to these food / beverage products can be selected according to the kind of food / beverage products used as the object of addition. However, if the amount added is too small, the expected anti-inflammatory effect, hyaluronidase inhibitory effect, and anti-allergic effect cannot be exhibited sufficiently. On the other hand, if the amount added is too large, the flavor inherent to the food to be added may be impaired. Therefore, it is preferable to determine the addition amount in consideration of these circumstances. From this viewpoint, the amount of the composition added is about 0.01% to 20%, preferably about 0.01% to 5% of the mass of the food or drink to be added.

  EXAMPLES Hereinafter, although an Example is shown and this invention is demonstrated concretely, this invention is not restrict | limited to each following example at all.

Example 1 Synthesis of Chlorogenic Acid Ethyl Ester 50 mL of ethanol, 0.1 g of reagent grade chlorogenic acid, and 1 mL of dilute sulfuric acid were mixed and heated and reacted at 40 ° C. for 3 hours under dark conditions. 50 mL of ethyl acetate and 50 mL of purified water were added and stirred, and the ethyl acetate layer was collected. The ethyl acetate layer was washed three times with purified water, the solvent of the obtained ethyl acetate layer was distilled off, and the concentrate was freeze-dried to introduce an ethyl group at the R ¹ position in the chemical formula (1). Chlorogenic acid ethyl ester was synthesized. The yield was 48.6%.
Confirmation of chlorogenic acid ethyl ester LC / MS Direct injection into MS confirms ESI + hydrogen addition ion 383.1 ESI- confirms hydrogen desorption ion 381.1 NMR (100 MHz, CD3OD)
13.0, 36.4, 36.6, 36.6, 61.3, 69.0, 70.8, 71.2, 74.4, 113.7, 113.7, 115.2, 121.7, 126.3, 145.5, 145.9, 148.4, 167.0, 173.6

[Example 2] Synthesis of 2-ethylhexyl chlorogenic acid 125 mL of 2-ethylhexanol, 0.1 g of reagent grade chlorogenic acid and 1 mL of dilute sulfuric acid were mixed, and heated and reacted at 40 ° C for 3 hours under dark conditions. 50 mL of ethyl acetate and 50 mL of purified water were added and stirred, and the ethyl acetate layer was collected. The ethyl acetate layer was washed 3 times with purified water, the solvent of the obtained ethyl acetate layer was distilled off, and the concentrate was lyophilized to give a 2-ethylhexyl group at the R ¹ position in chemical formula (1). The introduced chlorogenic acid 2-ethylhexyl ester was synthesized. The yield was 42.8%.
Confirmation of ethylhexyl chlorogenic acid LC / MS Direct injection into MS confirms 467.2 which is ESI + hydrogen addition ion ESI- Confirms 465.2 which is hydrogen desorption ion NMR (100 MHz, C5D5N)
10.8, 10.9, 14.0, 23.0, 23.7, 28.9, 28.9, 30.3, 30.4, 38.5, 38.8, 67.4, 67.4, 72.0, 114.9, 115.6, 116.5, 121.9, 126.6, 145.9, 147.5, 150.4, 166.8, 174.5

"Example 3" Synthesis of chlorogenic acid acetyl ester 0.1 g of reagent grade chlorogenic acid and 1 mL of acetic anhydride-pyrimidine mixed liquid (1: 2) were mixed and reacted at room temperature for 12 hours under dark conditions. After stirring 10 mL of purified water and 10 mL of ethyl acetate, the ethyl acetate layer was collected. The ethyl acetate layer was washed three times with purified water, the solvent of the obtained ethyl acetate layer was distilled off, and the concentrate was freeze-dried to synthesize chlorogenic acid acetyl ester. The yield was 68.6%.
Confirmation of acetylchlorogenic acid LC / MS Direct injection into MS confirms ESI + hydrogen addition ion 565.0 ESI- confirms hydrogen elimination ion 563.0 NMR (100 MHz, CD3OD)
19.1, 19.1, 19.3, 19.6, 19.9, 31.3, 36.4, 67.0, 68.1, 71.6, 79.1, 118.1, 122.9, 123.9, 126.4, 133.1, 142.8, 143.9, 144.1, 165.7, 168.3, 168.5, 170.2, 170.3, 170.4, 172.2

"Hyaluronidase inhibitory action"
Commercially available hyaluronic acid potassium salt (derived from human umbilical cord) was dissolved in 0.1 M phosphate buffer (pH 7.0) to a concentration of 0.9 mg / mL to obtain a substrate solution. A commercially available hyaluronidase (derived from bovine testis) was dissolved in 0.1 M phosphate buffer (pH 7.0) so as to be 5,3000 units / mL to obtain an enzyme solution. The enzyme solution was prepared at the time of use. In a test tube, 0.1 mL of a sample solution adjusted to each concentration with a buffer solution and 0.03 mL of an enzyme solution were taken and reacted at 37 ° C. for 20 minutes. Next, 0.06 mL of an activator was added and reacted at 37 ° C. for 20 minutes. Further, 0.15 mL of the substrate solution was added and reacted at 37 ° C. for 1 hour. 0.06 mL of 0.4N NaOH was added to stop the reaction, and the mixture was immediately cooled with ice. Then, 0.06 mL of borate buffer (pH 9.1) was added, and the mixture was boiled for 3 minutes and further cooled with ice. After 2.0 mL of p-dimethylbenzaldehyde (p-DABA) solution was added and reacted at 37 ° C. for 20 minutes, the solution was transferred from each test tube to a 96-well microplate, and the absorbance at 585 nm was measured using a microplate reader. It was measured. As a control, a solution to which only the buffer solution used for dissolving the sample was added was used. When the activity of hyaluronidase is inhibited, the degradation product N-acetylglucosamine decreases. Absorbance by p-DABA is lowered. Utilizing this fact, the inhibitory activity was obtained from the following equation. The results are summarized in Table 2.
Inhibition rate (%) = (control absorbance−sample absorbance) / control absorbance × 100

  As shown in Table 1, Examples 1 to 3 of the present invention showed high hyaluronidase inhibitory action. In contrast, chlorogenic acid did not have a significant hyaluronidase inhibitory effect. Therefore, the compound represented by the chemical formula (1) of the present invention exhibits a high hyaluronidase inhibitory action and is useful as an anti-inflammatory agent, a hyaluronidase inhibitor, and an antiallergic agent.

Claims (3)

In the compound represented by the following chemical formula (1),

R ¹ is an ethyl group or a 2-ethylhexyl group, an anti-inflammatory agent with one or the active ingredient R ² to R ⁶ are selected compound or we are H. A hyaluronidase inhibitor comprising the compound according to claim 1 as an active ingredient. An antiallergic agent comprising the compound according to claim 1 as an active ingredient.