JP7320897B1 - Pharmaceutical composition and food composition for enhancing uncoupling protein 2 gene expression - Google Patents

Abstract

[PROBLEMS] To provide a pharmaceutical composition for enhancing uncoupling protein 2 gene expression and a food composition for enhancing uncoupling protein 2 gene expression. SOLUTION: The present invention relates to a pharmaceutical composition for enhancing uncoupling protein 2 gene expression and a food composition for enhancing uncoupling protein 2 gene expression, containing 5-acetyl-1-benzylpyrrolidin-2-one as an active ingredient. . [Selection figure] None

Description

TECHNICAL FIELD The present invention relates to pharmaceutical compositions and food compositions for enhancing the expression of the uncoupling protein 2 (hereinafter simply referred to as "UCP2") gene.

5-Acetyl-1-benzylpyrrolidin-2-one, which is an active ingredient of the pharmaceutical composition and food composition according to the present invention, is the carbon at the 5-position of pyrrolidine. is attached to the acetyl group, the nitrogen at the 1-position is attached to the benzyl group, and the carbon at the 2-position is attached to the double bond of oxygen.

This 5-acetyl-1-benzylpyrrolidin-2-one is suggested as an example of a synthetic compound in Non-Patent Document 1 below, but its existence is only suggested, and no specific attribute is given. is neither disclosed nor suggested.

In addition, as shown in Patent Documents 1 to 5 below, the present inventors have already found that 5-acetyl-1-benzylpyrrolidin-2-one has PDE5 inhibitory action, estradiol production promoting action, muscle hypertrophy promoting action, and xanthine oxidase inhibitory action. and 5-aminolevulinic acid production-promoting action, but no action to enhance the expression of the UCP2 gene.

DESKUS, 2 others, "SYNTHESIS OF 5S-(1-OXOALKYL AND ARYL)-2-PYRROLIDINONE DERIVATIVES", SYNTHETIC COMMUNICATIONS, USA, Marcel Dekker, Inc., May 1998, Vol.28, No.9, p .1649-1659

Japanese Patent No. 6537689 Japanese Patent No. 6578046 Japanese Patent No. 6962630 Japanese Patent No. 7134541 Japanese Patent No. 7253301

As described above, there is no document disclosing or suggesting that 5-acetyl-1-benzylpyrrolidin-2-one has an effect of enhancing the expression of the UCP2 gene. It is known that the promotion of UCP2 production is caused by the UCP2 gene.

As a result of intensive research, the present inventors have found that 5-acetyl-1-benzylpyrrolidin-2-one has an action to enhance the expression of the UCP2 gene and further has an action to promote the production of UCP2. , have completed the present invention.

That is, the present invention relates to a pharmaceutical composition for enhancing UCP2 gene expression or a food composition for enhancing UCP2 gene expression, containing 5-acetyl-1-benzylpyrrolidin-2-one as an active ingredient.

The pharmaceutical composition and food composition for enhancing UCP2 gene expression of the present invention promote the production of UCP2 by enhancing the expression of the UCP2 gene. As a result, effects such as anti-inflammation by suppressing nitric oxide (NO) production, prevention and improvement of metabolic syndrome, prevention of heat stroke, and improvement of male infertility by suppressing decreased sperm motility can be expected.

The present invention relates to a pharmaceutical composition for enhancing UCP2 gene expression containing 5-acetyl-1-benzylpyrrolidin-2-one as an active ingredient. The pharmaceutical composition according to the present invention suppresses the increase in active oxygen (antioxidant) by promoting UCP2 production based on the enhanced expression of the UCP2 gene, and furthermore, anti-inflammatory by suppressing NO production, prevention and improvement of metabolic syndrome, and prevention of heat stroke. , Contributes to improving male infertility by suppressing decreased sperm motility.

5-acetyl-1-benzylpyrrolidin-2-one has an acetyl group attached to the 5-position carbon of pyrrolidine, a benzyl group attached to the 1-position nitrogen, and an oxygen to the 2-position carbon. is a compound having a double bond, and its structural formula is as shown in Formula 1 below. Also, _{the molecular} formula is _C13H15NO2 .

UCP is a protein that exists in the inner membrane of mitochondria and acts as a proton transporter, and uncouples the electron transport chain and ATP (adenosine triphosphate) synthesis by eliminating this potential gradient. Five isoforms (UCP1 to 5) are known to exist at present, and UCP2 is one of them, and is present in almost all tissues including skeletal muscle.

The pharmaceutical composition according to the present invention promotes the production of UCP2 by enhancing the expression of the UCP2 gene, which is the gene encoding UCP2, and suppresses the increase in active oxygen (antioxidant), and in turn, anti-inflammatory by suppressing NO production. , prevention and improvement of metabolic syndrome, prevention of heat stroke, and improvement of male infertility by suppressing decreased sperm motility.

The pharmaceutical composition according to the present invention can be used as a drug or quasi-drug, and can be in various forms. These pharmaceuticals or quasi-drugs are, for example, powders, pills, tablets (e.g., coated tablets, sugar-coated tablets, chewable tablets, etc.), capsules (e.g., hard or soft gelatin capsules, etc.), granules, oral liquids ( For example, it can be a dosage form suitable for oral administration such as an emulsion, suspension, syrup, etc.).

In addition, for example, dosage forms suitable for rectal administration such as suppositories, dosage forms suitable for intravascular administration such as injections and infusions, intramuscular administration, subcutaneous or intracutaneous administration, such as ointments, creams, Formulations suitable for topical or transdermal administration, such as gels or liquids (including eye drops, eye washes, etc.), that is, formulations suitable for parenteral administration, are preferred. Oral administration.

When the pharmaceutical composition of the present invention is used in the form of tablets, granules, capsules or chewable tablets, known carriers such as tableting aids, granule processing aids, capsule processing aids and the like are used. obtain.

Tableting aids include granulated sugar, refined sugar, powdered sugar, reduced maltose starch syrup powder, lactose, glucose, pullulan, erythritol, starch, dextrin, and other sugars, and dietary fibers such as crystalline cellulose, gum arabic, and bean curd refuse. , corn protein, food calcium such as calcium phosphate, food extracts, dried food powders, natural fruit juice powder, surfactants such as sucrose fatty acid esters, powdered oils and fats, glycerin, oils and fats such as fatty acid esters, or chewable tablets Flavoring materials such as various sweeteners, various acidulants, various flavors, shellac as coating materials, corn protein, yeast cell wall, starch, reduced maltose starch syrup, sugarless coating, maltitol, glycerin, sorbitol, HPMC, HPC etc. are exemplified. However, there is no particular limitation as long as the effects of the present invention are not impaired.

Also, the granule processing aid is not particularly limited as long as it does not impair the effects of the present invention, but granulated sugar, refined sugar, powdered sugar, reduced maltose starch syrup powder, lactose, glucose, pullulan, erythritol, starch, dextrin, etc. Examples include all sugars, dietary fibers such as crystalline cellulose and gum arabic, corn protein, food calcium such as calcium phosphate, food extracts, food dry powders, and natural fruit juice powders.

In addition, the capsule processing aid is not particularly limited as long as it does not impair the effects of the present invention. Glucose, pullulan, erythritol, starch, dextrin and other sugars, crystalline cellulose, gum arabic and other dietary fibers, corn protein, calcium phosphate and other food calcium, food extracts, food dry powders, natural fruit juice powders, etc. are contained in soft capsules. Content viscosity modifiers such as food oils and fats, beeswax, glycerin fatty acid esters, etc., for preparing capsules of this type are exemplified, respectively.

Tablets can usually be prepared using a tablet press, but tablets can also be blended with flavorings to make chewable tablets, or the surface of tablets can be coated using an automatic coating machine, a spray granulator, or a hand pan. You can Various granulators, such as spray granulator type, kneading (extrusion) type, or high speed agitation granulator type, can be used to form granules. Capsule filling machines (hard type and soft type) can be used to prepare capsules by mixing capsule aids.

The present invention also relates to a food composition for enhancing UCP2 gene expression, containing 5-acetyl-1-benzylpyrrolidin-2-one as an active ingredient. The food composition according to the present invention suppresses the increase in active oxygen (antioxidant) by promoting UCP2 production based on UCP2 gene expression enhancement, and furthermore, anti-inflammatory by suppressing NO production, prevention and improvement of metabolic syndrome, prevention of heat stroke, Effects such as improving male infertility by suppressing decreased sperm motility can be expected.

The food composition for enhancing UCP2 gene expression of the present invention can be used as food, beverage or animal feed, for example, health food, functional food, food for specified health use, beauty food or nutritional supplement (supplement). can. These foods, beverages and animal feeds may be, for example, in the form of known food such as ice cream, jelly, candy, chocolate and chewing gum, or in the form of drinking water such as tea and juice. It may also be in the form of liquids, powders, granules, capsules, tablets, or the like.

The intake of the pharmaceutical composition for enhancing UCP2 gene expression and the food composition for enhancing UCP2 gene expression according to the present invention is not particularly limited, but the age, body weight, symptoms, etc. It can be appropriately selected according to the dosage form. In addition, the period of ingestion can be arbitrarily determined according to the age and symptoms of the ingester or ingested animal.

EXAMPLES Next, the present invention will be described in more detail with reference to examples, but the present invention is not limited to the following examples as long as the gist thereof is not exceeded.

<Measurement of UCP2 gene expression level by DNA microarray>
In this test, after culturing human liver-derived cells Hep G2 in the presence of 5-acetyl-1-benzylpyrrolidin-2-one, DNA microarray analysis was performed to confirm whether there is an effect of enhancing UCP2 gene expression. .

<Liver-derived cells>
Human liver cancer line Hep G2
Obtained from the National Institute of Biomedical Innovation, Health and Nutrition.

<Preparation of specimen>
1) 100 μL of DMSO was added to 10 mg of 5-acetyl-1-benzylpyrrolidin-2-one to dissolve, and serially diluted with DMSO so that the concentration of the test substance was 50 mM.
2) Further, the test substance was diluted with a cell culture medium to a concentration of 50 μM.

<Method>
<Preparation of Hep G2 cells>
1) Hep G2 cells were seeded at a cell concentration of 2×10 ⁵ cells/mL in 5 mL each on a 6 cm dish.
2) Adhesion was confirmed 24 hours after seeding, and after removing the cell culture medium, 5 mL each of new cell culture medium containing the test substance (final concentration: 50 μM) was added.
3) After culturing for 24 hours after the addition, the cell culture medium was removed, the adherent cells were washed with PBS, and the obtained cells were used for the following RNA extraction→DNA microarray analysis.

<RNA extraction>
1) RNA was extracted from the cells obtained by the cell preparation described above using RNAiso plus and purified using the RNeasy MinElute Cleanup Kit.

<DNA microarray analysis>
1) RNA samples were synthesized using Low Input Quick Amp Labeling Kit (Agilent) to synthesize cDNA, and to synthesize and purify Cy3-labeled cRNA.
2) Using Gene Expression Hybridization Kit (Agilent), each labeled cRNA was fragmented, applied to Whole Human Genome Microarray Ver2.0 (Agilent), and hybridized at 65°C for 17 hours.
3) Array slides were washed using Gene Expression Wash Buffers 1 and 2 (Agilent).
4) Array images scanned with a microarray scanner (GenePix 4000B, Molecular Devices) were digitized with array analysis software GenePix Pro (Molecular Devices). The fluorescence intensity values were normalized, and the ratio of the test substance-added group to the control was calculated.

<result>
The expression of the UCP2 gene was enhanced 1.54-fold in the human liver-derived cell line (HepG2) in the presence of 5-acetyl-1-benzylpyrrolidin-2-one. Therefore, 5-acetyl-1-benzylpyrrolidin-2-one is considered to promote UCP2 production by enhancing UCP2 gene expression.

Claims (2)

A pharmaceutical composition for enhancing uncoupling protein 2 gene expression, comprising 5-acetyl-1-benzylpyrrolidin-2-one as an active ingredient. A food composition for enhancing uncoupling protein 2 gene expression, comprising 5-acetyl-1-benzylpyrrolidin-2-one as an active ingredient.