JP2016204363A - NF-κB ACTIVATION INHIBITOR - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain preparation which inhibits expression of NF-κB gene, inhibits formation of a crease, prevents a reduction in springiness, prevents skin sagging, and is effective as therapeutic agents for chronic inflammatory diseases, autoimmune diseases, viral diseases, immune diseases, and cancer and as prophylactic or therapeutic agents for other diseases caused by NF-κB activation.SOLUTION: The inventors have found out that blending extract of mulberry branches as an active ingredient serves the purpose of the invention.SELECTED DRAWING: None

Description

  The present invention suppresses NF-κB activation, suppresses skin wrinkles, prevents a decrease in elasticity, and prevents skin talmi. Furthermore, the present invention relates to a preparation effective as a preventive or therapeutic agent for chronic inflammatory diseases, autoimmune diseases, viral diseases, immune diseases, novel cancer treatments and other diseases caused by NF-κB activation.

NF-κB exists as a dimer formed from various combinations of p50, p65 / RelA, c-Rel, Rel-B and p52 which are members of the NF-κB family. Of these, the best known dimer is a heterodimer consisting of a 50 kDa subunit (p50) and a 65 kDa subunit (p65).
The heterodimer normally exists in an inactive state by binding to inhibitor of NF-κB (IκB) in the cytoplasm. However, when the cell is stimulated by inflammatory cytokines or cell growth factors, AKT IκB kinase is activated through a signal transduction pathway or the like, which causes phosphorylation of IκB, and phosphorylated IκB undergoes ubiquitination and is degraded by the proteasome. As a result, NF-κB is separated from IκB. It translocates into the nucleus and binds to the NF-κB response element to activate transcription of various target genes.

Many of these target genes are involved in inflammation and immune response, and NF-κB activation is associated with diseases such as rheumatoid arthritis, osteoarthritis, inflammatory bowel disease, atopic dermatitis, asthma, etc. Known to be involved in.
Moreover, it is known that various viruses such as HIV activate NF-κB in host cells, and it is considered that NF-κB contributes to virus infection.

  Furthermore, in recent years, it is known that NF-κB is often constitutively activated in various tumors. It is thought that it may be involved in the induction of the expression of various genes related to the development of the drug and resistance to anticancer drug treatment.

It is also known that NF-κB is involved in diseases such as ischemic heart disease, Alzheimer's disease, sepsis and metabolic syndrome.
Furthermore, overexpression of NF-κB is known to significantly age the skin, such as promoting MMP-1 production in the skin and degrading collagen.
Therefore, compounds that inhibit NF-κB are useful as preventive or therapeutic agents for chronic inflammatory diseases, autoimmune diseases, viral diseases, immune diseases, novel cancer treatments, and other diseases caused by NF-κB activation. Since the degradation of collagen is suppressed, NF-κB inhibitors have been actively developed. (Non-patent documents 1 to 11)

NF-κB is also activated by the presence of advanced glycation end products (AGE).
AGE is a substance produced by aging, which deposits on the skin, causing dullness and darkening and loss of skin transparency.
AGE is thought to enhance intracellular oxidative stress via RAGE (Receptor for AGE) and activate transcription factor NF-κB via the Ras / MAP kinase pathway in an oxidative stress-dependent manner.
Thus, AGE also activates NF-κB, but it is known that AGE is generated and accumulated with aging in human blood vessels, metabolic syndrome, diabetes, hyperglycemia, impaired glucose tolerance Inhibition of NF-κB activation is also known to be effective in preventing or ameliorating vascular disorders caused by hyperlipidemia, hypertriglyceremia, hypercholesterolemia, hypertension, obesity, arteriosclerosis, etc. Yes. (Patent Document 1)

JP 2007-204368 A Am. J. Respir. Cell Mol. Biol. 1997, 17, 3-9 N. Engl. J. Med. 1997, 336, 1066-1071 Nature 1987, 326, 711-713 Semin. Cancer Biol. 1997, 8, 121-129 Oncogene 1999, 18, 6938-6947 Cell Death Differ. 2006, 13, 738-747 Nat Med. 1997, 3, 894-899 J. Neural Transm. Suppl. 1997, 49, 125-134 J Crit Care. 1995, 10, 198-212 Obes Res. 2004, 12, 180-186. Tanaka K, Hasegawa J, Asamitsu K, and Okamoto T: J Pharmacol Exp Ther315: 624-630,2005

  The object of the present invention is to suppress the activation of NF-κB, to suppress skin wrinkles, to prevent the decrease in elasticity and to prevent the skin talmi. Furthermore, it is to obtain a preparation effective as a preventive or therapeutic agent for chronic inflammatory diseases, autoimmune diseases, viral diseases, immune diseases, novel cancer treatments and other diseases caused by NF-κB activation.

As a result of intensive studies by the present inventors, it has been found that the extract of mulberry branch has an action of suppressing the activity of NF-κB.
In addition, the collagen which caused Maillard reaction (hereinafter referred to as AGEs collagen) was added, and the NF-κB activation inhibitory effect was confirmed using fibroblasts that promoted NF-κB activity.
Details are described below.
In consideration of extraction efficiency, mulberry branches are preferably extracted after being subjected to processing such as chopping, drying, and pulverization.
Drying may be performed in the sun or using a commonly used dryer.
As the solvent used for the extraction, water, a hydrophilic organic solvent, or a mixture thereof is used.
Examples of water that can be used as the extraction solvent include pure water, tap water, well water, mineral spring water, mineral water, hot spring water, spring water, fresh water, and the like, as well as water that has been subjected to various treatments. Examples of the treatment applied to water include purification, heating, sterilization, filtration, ion exchange, adjustment of osmotic pressure, buffering, and the like. Therefore, the water that can be used as the extraction solvent in the present invention includes purified water, hot water, ion-exchanged water, physiological saline, phosphate buffer, phosphate buffered saline, and the like.
Examples of the hydrophilic organic solvent include lower alcohols having 1 to 5 carbon atoms such as methanol, ethanol, propyl alcohol, and isopropyl alcohol; lower aliphatic ketones such as acetone and methyl ethyl ketone; 1,3-butylene glycol, propylene glycol, Examples thereof include polyhydric alcohols having 2 to 5 carbon atoms such as glycerin, and a mixed solvent of these hydrophilic organic solvents and water can be used.
In addition, when using the mixed solvent of the said water and a hydrophilic organic solvent, in the case of a lower alcohol, it is 1-20 mass parts with respect to 10 mass parts of water, and in the case of a lower aliphatic ketone, it is 10 mass parts of water. On the other hand, it is preferable to add 1 to 15 parts by mass. In the case of a polyhydric alcohol, it is preferable to add 1 to 20 parts by mass with respect to 10 parts by mass of water.
The amount of the organic solvent used for the extraction is preferably about 5 to 100 times, more preferably about 10 to 50 times the amount of the plant (seaweed) as the raw material. Furthermore, in order to raise extraction efficiency, you may stir or homogenize in an extraction solvent. The extraction temperature is suitably about 5 ° C. to the boiling point of the extraction solvent. The extraction time varies depending on the type of extraction solvent and the extraction temperature, but it is appropriate to set it to about 1 hour to 14 days.
The extraction operation is not limited to a single operation. A fresh solvent can be added again to the residue after extraction to perform an extraction operation, or the extraction solvent can be brought into contact with the extraction raw material a plurality of times.
If necessary, purification treatment such as deodorization and decolorization may be further added within a range that does not affect the effect, and the concentration can be achieved by a vacuum concentrator such as an evaporator or solvent removal by heating.
Further, this extract can be further purified using a synthetic adsorbent (Diaion HP20, Sephabies SP825, Amberlite XAD4, MCIgelCHP20P, etc.), dextran resin (Sephadex LH-20, etc.), ultrafiltration and the like. is there.

In addition to these extracts, it is possible to prepare a preparation together with raw materials that can be used for pharmaceuticals, foods, and cosmetics, or to add these to existing pharmaceuticals, foods, and cosmetics.
Examples of available raw materials
Avocado oil, almond oil, fennel oil, egoma oil, olive oil, orange oil, orange rafa oil, sesame oil, cacao butter, chamomile oil, carrot oil, cucumber oil, beef tallow fatty acid, kukui nut oil, safflower oil, shea butter, liquid shea Fat, soybean oil, camellia oil, corn oil, rapeseed oil, persic oil, castor oil, cottonseed oil, peanut oil, turtle oil, mink oil, egg yolk oil, palm oil, palm kernel oil, owl, coconut oil, beef tallow, lard , Squalene, squalane, pristane or various fats and oils such as hydrogenated products (hardened oil, etc.) of these fats and oils.
Waxes such as beeswax, carnauba wax, whale wax, lanolin, liquid lanolin, reduced lanolin, hard lanolin, candelilla wax, montan wax, shellac wax, rice wax;
Mineral oils such as liquid paraffin, petrolatum, paraffin, ozokelide, ceresin, and microcristan wax.

Fatty acids such as lauric acid, myristic acid, palmitic acid, stearic acid, behenic acid, oleic acid, linoleic acid, linolenic acid, docosahexaenoic acid, eicosapentaenoic acid, 12-hydroxystearic acid, undecylenic acid, tall oil, and lanolin fatty acid.
Alcohols such as ethanol, isopyropanol, lauryl alcohol, cetanol, stearyl alcohol, oleyl alcohol, lanolin alcohol, cholesterol, phytosterol, phenoxyethanol, 2-hexyldecanol, isostearyl alcohol, 2-octyldodecanol.
Ethylene glycol, diethylene glycol, triethylene glycol, ethylene glycol monoethyl ether, ethylene glycol monobutyl ether, diethylene glycol monomethyl ether, diethylene glycol monoethyl ether, polyethylene glycol, propylene glycol, polypropylene glycol, 1,3-butylene glycol, pentyl glycol, glycerin, Polyhydric alcohols such as pentaerythritol, threitol, arabitol, xylitol, galactitol, sorbitol, lactitol, and maltitol.

Isopropyl myristate, isopropyl palmitate, butyl stearate, hexyl laurate, myristyl myristate, oleyl oleate, decyl oleate, octyldodecyl myristate, hexyldecyl dimethyloctanoate, cetyl lactate, myristyl lactate, diethyl phthalate, phthalate Esters such as dibutyl acid, lanolin acetate, ethylene glycol monostearate, propylene glycol monostearate, propylene glycol dioleate.
Metal soaps such as aluminum stearate, magnesium stearate, zinc stearate, calcium stearate and zinc palmitate.
Gum arabic, guaiac fat, karaya gum, tragacanth gum, quince seed, agar, casein, lactose, fructose, sucrose or ester thereof, trehalose or derivative thereof, dextrin, gelatin, pectin, starch, carrageenan, carboxymethyl chitin or chitosan, ethylene oxide Hydroxyalkyl (C2 to C4) chitin or chitosan to which alkylene (C2 to C4) oxide is added, etc., low molecular chitin or chitosan, chitosan salt, sulfated chitin or chitosan, phosphorylated chitin or chitosan, alginic acid or a salt thereof, Hyaluronic acid or its salt, chondroitin sulfate or its salt, heparin, ethyl cellulose, methyl cellulose, carboxymethyl cellulose, carboxyethyl cellulose, carboxyethyl cellulose sodium, hyaluronan Droxyethyl cellulose, hydroxypropyl cellulose, nitrocellulose, crystalline cellulose, polyvinyl alcohol, polyvinyl methyl ether, polyvinyl pyrrolidone, polyvinyl methacrylate, polyacrylate, polyalkylene oxide such as polyethylene oxide and polypropylene oxide, or a crosslinked polymer thereof, carboxy Gum, saccharides or water-soluble polymer compounds such as vinyl polymer and polyethyleneimine.
Alkyl carboxylate, alkyl sulfonate, alkyl sulfate ester salt, alkyl phosphate ester salt, alkyl amine salt, alkyl quaternary ammonium salt, carboxylic acid type amphoteric surfactant, sulfate ester type amphoteric surfactant, sulfonic acid type Amphoteric surfactant, phosphate ester type amphoteric surfactant, ether type nonionic surfactant, ether ester type nonionic surfactant, ester type nonionic surfactant, block polymer type nonionic surfactant, nitrogen-containing Anionic surfactants such as type nonionic surfactants, cationic surfactants, amphoteric surfactants, nonionic surfactants.

Retinol, retinal, dehydroretinal, carotene, lycopene, thiamine hydrochloride, thiamine sulfate, riboflavin, pyridoxine, cyanocobalamin, folic acid, nicotinic acids, pantothenic acids, biotins, choline, inositols, vitamin C or derivatives thereof, ergocalci Ferrol, cholecalciferol, dihydrotaxosterol, vitamin E or its derivatives, various vitamins such as ubiquinones or their derivatives.
Valine, leucine, isoleucine, threonine, methionine, phenylalanine, tryptophan, lysine, glycine, alanine, asparagine, glutamine, serine, cysteine, cystine, tyrosine, proline, hydroxyproline, aspartic acid, glutamic acid, hydroxylysine, ornithine, histidine, etc. And various amino acids such as their sulfates, phosphates, nitrates, citrates, or amino acid derivatives such as pyrrolidone carboxylic acid.

  Red buds, ground yellow, woody, gallbladder, monomedicine, ginkgo, oolong tea, fennel, turmeric, summer hay, Chen bark, Ezokogi, Akemiko, auren, white birch, giant crape myrtle, okra, ginseng or tochibaninjin (carrot), olive, oregano , Orange, chamomile or roman chamomile, cam come, guarana, magnolia, gorye, gooseberry, clara, cranberry, grapefruit, clove, kobushi, magnolia, mausoleum, sakura, hemp, sanchinin carrot, yam, yam, sandscon, Shikunshi, shiso shiso, shiso, peony, honeysuckle, horseradish, celery, cucumber, yellowtail, daisou, chimpanzee, chikutsujinjin, tengusa, mocha tea, ladybird, toxa, dokudami, tochu, ash, cinnamon, cinnamon Double, Neubara, Novara, Barley, Rose, Rhizome, Loquat, Blackberry, Prunes, Hornbill, Bowfish (Windbreak), Honoki, Bodaiju, Button, Hop, Jojoba, Maow, Maca, Macadamia nut, Mulberry bark, Bitter , Garlic mushroom, berry, milobaran, mugwort, purple, beetle grass, evening primrose, melissa, melirot, mokko, yacon, eucalyptus, saxifrage, yucca, yuzu, lily, licorice, mugwort, lavender, lemon, lemongrass, rosemary, earthenware, Various extracts of plants, seaweeds, and algae such as chlorella, chlorella, kombu, wakame, giant kelp, hijiki, hibamata, giraffe, asakusanori, susbinori, spirulina, shigege, iroro, fukuronori.

Horse or pig placenta extract, silk protein and its degradation products or their derivatives, milk, casein and its degradation products or their derivatives, skim milk powder and its degradation products or their derivatives, lactoferrin or its degradation products, egg component Animal-derived materials such as fish meat degradation products and nucleic acid-related substances (ribonucleic acid, deoxyribonucleic acid).
Marine products such as deep sea water, sea salt, dried sea water, dead sea or Atlantic or Pacific ocean, marine products such as sea mud.
Yeast extract, bacterial metabolite, bacterial extract, mold or actinomycete metabolite, mold or actinomycete extract, natto metabolite, natto extract, rice fermented extract, rice bran (red koji, birch) fermented extract, Fungi-derived substances such as lactic acid fermentation products of raw milk or skim milk powder, lactic acid bacteria fermentation products of legumes, and lactic acid bacteria fermentation products of coconut palm plants.
Α-hydroxy acids such as glycolic acid, citric acid, malic acid, tartaric acid and lactic acid.
Silica, magnesium silicate, talc, kaolin, bentonite, mica, mica titanium, bismuth oxychloride, zirconium oxide, magnesium oxide, zinc oxide, titanium oxide, calcium carbonate, magnesium carbonate, yellow iron oxide, red iron oxide, black iron oxide , Inorganic pigments such as Gunjo, chromium oxide, chromium hydroxide, carbon black, and calamine.
UV absorbers or blocking agents such as benzophenone derivatives, paraaminobenzoic acid derivatives, methoxycinnamic acid derivatives, salicylic acid derivatives, urocanic acid derivatives.

Paraaminobenzoic acid derivative, salicylic acid derivative, anthranilic acid derivative, coumarin derivative, amino acid compound, benzotriazole derivative, hydroquinone or derivative thereof, nicotinic acid derivative, kojic acid or derivative thereof, oxybenzone, benzophenone, arbutin, guaiazulene, shikonin, baicalin , Whitening agents such as baicalein, berberine, placenta extract, ellagic acid, lucinol.
Ictamol, indomethacin, kaolin, salicylic acid, sodium salicylate, methyl salicylate, acetylsalicylic acid, diphenhydramine hydrochloride, d-camphor, dl-camphor, hydrocortisone, guaiazulene, camazulene, chlorpheniramine maleate, glycyrrhizic acid or its salt, glycyrrhetinic acid or its Salt anti-inflammatory agent.
Antibacterial, bactericidal and antiseptics such as acrinol, sulfur, calcium gluconate, chlorhexidine gluconate and triclosan.
Musk, Civet, Castorium, Ambergris, Ylang Ylang Essential Oil, Iris Essential Oil, Fennel Essential Oil, Orange Essential Oil, Cardamom Essential Oil, Cinnamon Essential Oil, Geranium Essential Oil, Copaiba Balsam Essential Oil, Cedarwood Essential Oil, Jasmine Essential Oil, Rose Essential Oil, Bergamot Essential Oil, Lavender Essential Oil, Animal and vegetable fragrances such as lemongrass essential oil, lemon essential oil, rosemary essential oil, and other synthetic fragrances.
Photosensitive element 101, photosensitive element 201, photosensitive element 401, photosensitive element 301, hinokitiol, pantothenic acid or derivatives thereof, allantoin, pentadecanoic acid glyceride, urea, guanidine and other various drugs.
Other hormones, sequestering agents, pH adjusters and the like can be mentioned.

From these, various selections are made and used in any form. Examples of forms include capsules, powders, granules, solids, liquids, gels, bubbles, emulsions, creams, ointments, sheets and the like.
The blending amount of the mulberry branch extract in the preparation is preferably 0.00001 to 10% by weight, more preferably 0.0001 to 1.0% by weight as the solid content.

Example 300 ml of 50% ethanol was added to 10 g of mulberry branch (dried product) and allowed to stand for 5 days with occasional stirring. This was filtered, evaporated, and lyophilized.

The following experiment was conducted on the examples.
Human normal fibroblasts (Kurabo) were seeded in a 6-well plate, and when confluent, AGE collagen was applied at 0.5 mg / ml and Example 0.5 mg / ml.
After acting for 5 hours, the cells were collected and the nuclei were extracted. Using the nuclear extract, the amount of NF-κB was measured using NF-κB (p65) Transcription factor assay kit (CayMan Chemical).
In addition, the test of the system which added AGE-ized collagen and a system which does not add any Example as a control | contrast, and the system which added only AGE-ized collagen was implemented.
In addition, the preparation method of AGEs collagen was performed as follows.
50 mg of collagen was dissolved in 1 mM HCl, and sodium cyanoborohydride (1.42 g) and glyoxylic acid (0.715 g) were added to PBS (25 ml).
After reacting at 37 ° C. for 24 hours, ultrafiltered (molecular weight 30000) and lyophilized were used.
The results are shown in FIG.

It is a result of an NF-κB activation inhibition test, and is a view showing that Examples inhibit NF-κB whose expression is increased by AGEs collagen depending on the concentration. In addition, the result was expressed as 1 indicating the expression level of NF-κB in an experiment in which AGEs collagen was added and no example was added.

Claims (1)

  NF-κB activation inhibitor containing mulberry extract