JP7646338B2 - Aquaporin gene expression promoter - Google Patents

Description

The present invention relates to an aquaporin gene expression promoter.

Aquaporins (AQPs) are proteins with pores that exist in cell membranes, and are called water channels because they selectively allow water molecules to pass through but do not allow ions or other substances to pass through (Non-Patent Document 1). Aquaporins form tetramers in cell membranes, and each monomer functions as a water channel. Different types of aquaporins have different peptide sequences, and the size of the channel pores varies accordingly. Small pores allow only small molecules such as water molecules to pass through, while blocking charged factors, thereby maintaining the electrochemical potential of the membrane.

In humans, 13 types of aquaporins (0 to 12) are known to exist, and the main expression sites, main functions and characteristics of each aquaporin have been identified.

Various ingredients that can regulate aquaporin function have been reported with the aim of controlling the actions of aquaporins based on their functions and characteristics. For example, a kumazasa extract preparation has been reported as an ingredient that promotes AQP1 expression (Patent Document 1), hyaluronic acid as an AQP2 function enhancer (Patent Document 2), saffron extract, royal jelly extract, butcher's broom extract, etc. as AQP3 production enhancers (Patent Documents 3 to 5), 2-guanidino-1-ethanesulfonic acid as an AQP4 function enhancer (Patent Document 6), retinoic acid or a salt thereof as an AQP5 production enhancer (Patent Document 7), and alpha-linolenic acid as an AQP7 expression enhancer.

Q. Rev. Biophys. 39 (4): 361-396, 2006

JP 2020-2054 A JP 2015-96493 A JP 2005-343882 A JP 2009-184955 A JP 2009-235044 A International Publication No. 2017/150704 JP 2007-332048 A JP 2018-203680 A

As such, various ingredients that can regulate the function of aquaporins have been reported, but considering the wide range of actions of aquaporins, new materials that can regulate the function of aquaporins are still needed.

The object of the present invention is to provide a new active ingredient that regulates the function of aquaporins.

After extensive research, the inventors discovered that Daisaikoto extract and Bofutsushosan extract, which were not previously known to be related to aquaporins, have the effect of promoting the expression of aquaporin genes. The present invention was completed based on this knowledge and through further research.

That is, the present invention provides the following aspects.
Item 1. An aquaporin gene expression promoter comprising Daisaikoto extract and/or Bofutsushosan extract.
Item 2. The aquaporin gene expression promoter according to Item 1, which is used to promote expression of the aquaporin 7 gene.

The present invention provides a new active ingredient for promoting aquaporin gene expression.

1 shows the promoting effect of AQP7 gene expression by Bofu-tsusho-san and Daisaiko-to.

1. Aquaporin Gene Expression Promoter The aquaporin gene expression promoter of the present invention is characterized by containing Daisaikoto extract and/or Bofutsushosan extract. The aquaporin gene expression promoter of the present invention will be described in detail below.

The active ingredient Bofutsushosan is preferably a Kampo prescription described in "New Guide to General Kampo Prescriptions" (edited by Goda Yukihiro and Hakamzuka Takashi, edited by the Japan Kampo Herbal Medicine Preparation Association, published by Jiho Co., Ltd.), which includes mixed herbal medicines consisting of Angelica Root, Peony Root, Cnidium Root, Sanshishi, Forsythia Fruit, Mint, Ginger, Cabbage, Scutellaria Root, Ephedra, Rhizome, Boswellia, Byakujutsu, Platycodon, Scutellaria Root, Licorice, Scutellaria Root, and Kasseki. Some letters do not contain Byakujutsu among the above-mentioned herbal medicines (for example, "Kenken Kampo Prescription Quantity Collection," supervised by Otsuka Keisetsu and Yakazu Domyo, published by Ido no Nihonsha), and some do not contain Scutellaria Root (for example, "Zoku Kampo Arekore," edited by Osaka Yomiuri Shimbun, published by Naniwasha). Additionally, some Bofu-tsusho-san prescriptions contain Glehnia Root instead of Bofu-fu, and some contain Sou-atracto Root instead of Byaku-atracto Root.

The amounts of each herb that makes up Bofu-tsusho-san are as follows: Angelica Root 1.2 parts by weight, Peony Root 1.2 parts by weight, Cnidium Root 1.2 parts by weight, Sanshi Fruit 1.2 parts by weight, Forsythia Fruit 1.2 parts by weight, Mint Root 1.2 parts by weight, Zingiber Root 0.3-1.2 parts by weight, Caulerpa Root 1.2 parts by weight, Bougainvillea Root 1.2 parts by weight, Ephedra Root 1.2 parts by weight, Rhubarb Root 1.5 parts by weight, Bougainvillea Root 1.5-3.0 parts by weight, Atractylodes Rhizome 2.0 parts by weight, Platycodon Root 2.0 parts by weight, Scutellaria Root 2.0 parts by weight, Licorice Root 2.0 parts by weight, Glycyrrhiza Root 2-3 parts by weight, and Glycyrrhiza Root 3-5 parts by weight.

As for Daisaikoto, the Kampo prescription described in the "New Guide to General Kampo Prescriptions" (supervised by Goda Yukihiro and Hakamzuka Takashi, edited by the Japan Kampo Herbal Medicine Preparation Association, published by Jiho Co., Ltd.) is preferable, and includes a mixture of herbs consisting of Saiko, Pinellia Root, Scutellaria Root, Pheasant's Root, Peony Root, Shokyo, Taisou, and Daio. In addition, Daisaikoto includes the mixture of herbs (Kampo prescriptions) described in currently used Kampo-related letters, as stipulated in the "Basic Handling Policy for Kampo Preparations" established by the Kampo Herbal Medicine Investigation Committee.

The amounts of each herb that makes up Daisaikoto include: Bupleurum Root 3-12 parts by weight, preferably 4-9 parts by weight; Pinellia Root 2-8 parts by weight, preferably 2.5-6 parts by weight; Scutellaria Root 1.5-6 parts by weight, preferably 2-4.5 parts by weight; Pheasant's Root 1-4 parts by weight, preferably 1.5-3 parts by weight; Peony Root 1.5-6 parts by weight, preferably 2-4.5 parts by weight; Shokyo Root 0.5-2 parts by weight, preferably 1-1.5 parts by weight; Taisou Root 1.5-6 parts by weight, preferably 2-4.5 parts by weight; and Daio Root 0.5-2 parts by weight, preferably 1-1.5 parts by weight.

The bofutsushosan extract and daisaikoto extract of the present invention may be obtained as an extract liquid by extracting the above-mentioned mixed herbal medicines and concentrating the obtained extract liquid as necessary, or may be obtained as an extract powder by drying the extract liquid.

In the production of Bofutsushosan extract and Daisaikoto extract, the extraction solvent used in the extraction process is not particularly limited, but examples include water or aqueous ethanol. The drying process is also not particularly limited, and includes known methods such as the spray drying method and a method in which a suitable adsorbent (e.g., silicic anhydride, starch, etc.) is added to a soft extract with an increased concentration of the extract liquid to produce an adsorbed powder.

The Bofutsushosan extract and Daisaikoto extract used in the present invention may be either extracts prepared by the above-mentioned method or commercially available. For example, Bofutsushosan extract powders include "Bofutsushosan Dried Extract A", "Bofutsushosan Dried Extract AM", "Bofutsushosan Dried Extract E", and "Bofutsushosan Dried Extract EM" (all manufactured by Nippon Powder Co., Ltd.), and "Bofutsushosan Dried Extract-C" and "Bofutsushosan Dried Extract-F" (all manufactured by Alps Pharmaceutical Co., Ltd.), which are known as commercial products and can be obtained commercially. Daisaikoto extract powders include Daisaikoto Dried Extract AM, Daisaikoto Dried Extract SN, and Daisaikoto Dried Extract Powder (all manufactured by Nippon Powder Co., Ltd.), as well as Daisaikoto Dried Extract F and Daisaikoto Dried Extract-F (all manufactured by Alps Pharmaceutical Co., Ltd.), which are known as commercial products and can be obtained commercially.

In the aquaporin gene expression promoter of the present invention, the content of the active ingredient, the herbal extract, is not particularly limited as long as the effect of the present invention is exhibited, but is usually 5 to 100% by weight, preferably 10 to 90% by weight, more preferably 20 to 80% by weight, and even more preferably 30 to 60% by weight, calculated as the amount of the dried extract powder of the active ingredient, the herbal extract. In the present invention, the amount of the dried extract powder of the active ingredient, the herbal extract, refers to the amount itself when the dried extract powder of the herbal extract is used, and refers to the amount remaining after removing the solvent when the liquid extract of the herbal extract is used. In addition, when the dried extract powder of the herbal extract contains additives such as adsorbents added during production, the amount refers to the amount excluding the additives.

Other components The aquaporin gene expression promoter of the present invention may be composed of the herbal extract as an active ingredient alone, or may contain additives and bases according to the formulation.Such additives and bases are not particularly limited as long as they are pharmacologic acceptable, and include, for example, excipients, binders, disintegrants, lubricants, isotonicity agents, plasticizers, dispersants, emulsifiers, solubilizers, wetting agents, stabilizers, suspending agents, adhesives, coating agents, glossing agents, water, oils and fats, waxes, hydrocarbons, fatty acids, higher alcohols, esters, water-soluble polymers, surfactants, metal soaps, lower alcohols, polyhydric alcohols, pH adjusters, buffers, antioxidants, UV inhibitors, preservatives, flavorings, fragrances, powders, thickeners, pigments, chelating agents, etc.These additives may be used alone, or in combination of two or more.In addition, the content of these additives and bases is appropriately set according to the type of additives and bases used, the formulation form of the aquaporin gene expression promoter, etc.

In addition, the aquaporin gene expression promoter of the present invention may contain other nutritional components or pharmacological components as necessary, in addition to the above-mentioned active ingredients. Such nutritional components and pharmacological components are not particularly limited as long as they are pharmacologic acceptable, and examples thereof include antacids, stomachic agents, digestive agents, intestinal regulators, antispasmodics, mucosal repair agents, anti-inflammatory agents, astringents, antiemetics, antitussives, expectorants, anti-inflammatory enzymes, sedatives, hypnotics, antihistamines, caffeine, cardiac diuretics, antibacterial agents, vasoconstrictors, vasodilators, local anesthetics, herbal extracts, vitamins, and menthols. These nutritional components and pharmacological components may be used alone or in combination of two or more. The content of these components is appropriately set depending on the type of components to be used.

The formulation form of the aquaporin gene expression promoter of the present invention is not particularly limited as long as it can be administered orally, and examples of the formulation include solid formulations such as powders, fine granules, granules (including dry syrup), tablets, pills, capsules (soft capsules, hard capsules), etc.; semi-solid formulations such as jellies; and liquid formulations such as solutions, suspensions, syrups, etc. Among these formulation forms, solid formulations are preferred from the viewpoints of the stability and portability of the contained ingredients.

To prepare the aquaporin gene expression promoter of the present invention in the above-mentioned formulation form, the daisaikoto extract and/or bofutsushosan extract, and additives, bases, and pharmacological ingredients added as necessary, may be formulated according to the usual formulation methods used in the pharmaceutical field.

Uses: Daisaikoto extract and/or Bofutsushosan extract, which are the active ingredients of the aquaporin gene expression promoter of the present invention, promote the expression of aquaporin genes. Therefore, the aquaporin gene expression promoter of the present invention is used for the purpose of promoting the expression of aquaporin genes. Particularly preferably, the aquaporin gene expression promoter of the present invention is used for the purpose of promoting the expression of aquaporin 7 gene.

The aquaporin gene expression promoter of the present invention can be used by people of all ages and genders, but is preferably used in postmenopausal and/or menopausal women.

Dosage and Usage: The aquaporin gene expression promoter of the present invention is administered orally. The dosage of the aquaporin gene expression promoter of the present invention is appropriately set depending on the age, sex, constitution, etc. of the subject of administration, but for example, it may be administered in an amount equivalent to about 1 to 15 g of Daisaikoto extract in terms of the amount of dried extract powder per person per day, or in an amount equivalent to about 1 to 15 g of Bofutsushosan extract in terms of the amount of dried extract powder per person, 1 to 3 times a day, preferably 2 or 3 times a day. There is no particular restriction on the timing of administration, and it may be administered before, after, or between meals, but it is preferable to administer it before meals (30 minutes before a meal) or between meals (2 hours after a meal).

In order to further enhance the effect of promoting aquaporin gene expression, it is preferable to take the drug continuously for, for example, one week or more, preferably four weeks or more, and more preferably seven weeks or more.

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

Preparation of Daisaikoto Extract Powder The raw herbs used were 6.0 (weight parts, the same below), 4.0, 1.0, 3.0, 3.0, 3.0, 2.0, and 1.0 of Saiko, Pinellia Root, and then chopped, and extracted with 20 times the weight (460 weight parts) of water at about 100°C for 1 hour, centrifuged to obtain an extract, which was concentrated under reduced pressure and dried using a spray dryer to obtain Daisaikoto Extract Powder. The obtained Daisaikoto Extract Powder was 2.4g per 13.8g of raw herb mixture. The drying using the spray dryer was performed by dropping the extract into an atomizer rotating at 10,000 rpm and supplying hot air at 150°C.

The raw herbs used for preparing the Bofutsushosan extract powder were 1.2 parts by weight (parts by weight, hereinafter the same), 1.2 parts by weight of Peony Root, 1.2 parts by weight of Cnidium Root, 1.2 parts by weight of Sanshishi, 1.2 parts by weight of Forsythia Fruit, 1.2 parts by weight of Mentha Root, 1.2 parts by weight of Zingiber officinale, 1.2 parts by weight of Scutellaria Baicalensis, 1.2 parts by weight of Scutellaria Baicalensis, 2.0 parts by weight of Licorice Root, 2.0 parts by weight of Gypsum Root, 2.0 parts by weight of Gypsum Root, and 3.0 parts by weight of Glycyrrhiza ...

Test Example: Female C57BL/6J mice were used, and ovariectomized at 6 weeks of age. After recovery for 1 week, they were fed with food (High Fat Diet 32, manufactured by CLEA Japan) for 10 days to prepare a model mouse with suppressed aquaporin genes in menopause. The model mice were divided into a group that received food containing 2% by weight of Bofutsushosan or Daisaikoto, and a group that received only food, and were raised for 7 weeks. The feed was given ad libitum. The average intake of the feed containing the Kampo and the feed not containing the Kampo was 3.9 g/mouse/day. After 7 weeks of raising, white adipocytes were extracted and RT-qPCR was performed to measure the expression level of the AQP7 gene. The group that received only food was used as a control, and a comparison was performed using the Dunnett method with the Bofutsushosan group and the Daisaikoto group. The results are shown in Figure 1.

As shown in Figure 1, Bofutsushosan and Daisaikoto significantly and remarkably promoted the expression of the AQP7 gene.

Claims (1)

An aquaporin gene expression promoter containing Daisaikoto extract and/or Bofutsushosan extract for use in promoting the expression of aquaporin 7 gene in postmenopausal women and/or menopausal women.