JP7582612B2 - Sodium-dependent glucose cotransporter inhibitors - Google Patents

Description

The present disclosure relates to sodium-dependent glucose cotransporter (SGLT) inhibitors.

In recent years, the number of diabetic patients has been steadily increasing due to the Westernization of dietary habits, high calorie intake, lack of exercise caused by the development of a car-oriented society, and the aging population. Diabetes is a systemic metabolic disorder that causes chronic hyperglycemia due to insufficient insulin action in the body caused by reduced insulin sensitivity in the liver, fat, and muscles, and reduced insulin secretion ability in the pancreas, and is known to promote the onset and progression of cardiovascular diseases such as stroke and ischemic heart disease. Diabetes not only reduces quality of life, but also places a huge burden on society in terms of medical and economic aspects, making it a social problem.

On the other hand, SGLT, which actively transports glucose in a sodium concentration-dependent manner, and glucose transporters (GLUTs), which passively transport glucose, have been identified as sugar transporters that take up glucose into the body. Of these sugar transporters, SGLT has three subtypes: SGLT1, SGLT2, and SGLT3. For example, SGLT1 is expressed in the small intestine and kidney, and is known to absorb glucose derived from food in the small intestine and reabsorb glucose from urine in the kidney. In recent years, attempts have been made to prevent or treat diabetes by using SGLT inhibitors to inhibit or delay glucose absorption, thereby suppressing the rise in blood glucose levels (see, for example, Patent Documents 1 to 3).

JP 2019-182852 A JP 2012-62308 A JP 2011-32174 A

Diabetic patients need to take SGLT inhibitors, including SGLT1, for the long term, so it would be desirable to be able to easily ingest SGLT inhibitors that use naturally occurring ingredients in the form of food or other products; however, most of the SGLT inhibitors reported so far are synthetic compounds that do not exist in nature. In addition, although chlorogenic acid, a naturally occurring ingredient, is known to have SGLT inhibitory activity, its SGLT inhibitory activity is not sufficient.

Therefore, the objective of this disclosure is to identify components with excellent SGLT inhibitory activity and provide new SGLT inhibitors.

The present inventors conducted intensive research to solve the above problems and found that pectolinarigenin and/or its glycosides have excellent SGLT inhibitory activity and can be used as active ingredients of SGLT inhibitors. The invention described in this disclosure was completed through further investigations based on this knowledge.

The present disclosure provides SGLT inhibitors according to the embodiments set out below.
Item 1. An SGLT inhibitor containing pectinarigenin and/or its glycoside as an active ingredient.
Item 2. The SGLT inhibitor according to Item 1, which is used for inhibiting SGLT.
Item 3. The SGLT inhibitor according to Item 1 or 2, which is used for the prevention or treatment of diabetes.
Item 4. The SGLT inhibitor according to any one of Items 1 to 3, which is a food or drink for inhibiting sodium-dependent glucose cotransporter.
Item 5. The SGLT inhibitor according to any one of Items 1 to 3, which is a pharmaceutical for inhibiting a sodium-dependent glucose cotransporter.

According to the SGLT inhibitor disclosed herein, by using pectinarigenin and/or its glycosides as active ingredients, SGLT can be effectively inhibited, making it possible to prevent or treat diseases and symptoms caused in part by hyperglycemia, such as diabetes. Furthermore, since pectinarigenin and its glycosides are naturally occurring ingredients, the SGLT inhibitor disclosed herein is highly safe and can be used in a variety of products, such as food and beverages and pharmaceuticals.

FIG. 1 shows the results of evaluating the effects of pectinarigenin and chlorogenic acid on SGLT1 activity in Test Example 1. FIG. 1 shows the results of evaluating the relationship between pectinarigenin concentration and SGLT1 inhibitory activity in Test Example 2. FIG. 1 shows the results of evaluating the effects of pectolinarigenin and pectolinarin on SGLT1 activity in Test Example 3.

One embodiment of the present disclosure is an SGLT inhibitor containing pectinarigenin and/or its glycoside as an active ingredient. The SGLT inhibitor that is one embodiment of the present disclosure is described in detail below.

[Active ingredient]
The SGLT inhibitors of the present disclosure use pectinarigenin and/or its glycosides as active ingredients.

Pectolinarigenin is a type of flavonoid also known as 5,7-dihydroxy-4',6-dimethoxyflavone (IUPAC name) and is the aglycone of pectolinarin.

A glycoside of pectinarigenin is a compound in which a sugar is glycosidically bonded to pectinarigenin. An example of a glycoside of pectinarigenin used in the SGLT inhibitor of the present disclosure is a compound in which a sugar is glycosidically bonded to the hydroxyl group of the A ring of pectinarigenin. In the glycoside of pectinarigenin, the type of sugar bound is not particularly limited, and examples thereof include monosaccharides such as glucose, mannose, galactose, rhamnose, xylose, arabinose, apiose, etc.; disaccharides consisting of one or two of these monosaccharides; trisaccharides consisting of one or more of these monosaccharides; tetrasaccharides consisting of one or more of these monosaccharides; and pentasaccharides consisting of one or more of these monosaccharides. In one embodiment of the SGLT inhibitor of the present disclosure, a suitable example of a glycoside of pectinarigenin is pectolinarin.

In the SGLT inhibitor of the present disclosure, one compound selected from pectinarigenin and its glycosides may be used as the active ingredient, or two or more compounds may be used in combination. In one embodiment of the SGLT inhibitor of the present disclosure, the active ingredient used is preferably pectinarigenin and/or pectolinarin, more preferably pectinarigenin.

The origin of pectolinarigenin and/or its glycosides used in the SGLT inhibitors disclosed herein is not particularly limited, and they may be obtained from plants or by chemical synthesis, enzymatic synthesis, etc.

From the viewpoints of safety, reduction of manufacturing costs, etc., one embodiment of the SGLT inhibitor of the present disclosure can use pectinarigenin and/or its glycosides derived from plants. Examples of plants from which pectinarigenin is derived include plants of the genus Cirsium in the family Asteraceae, specifically Cirsium brevicaule, Cirsium boninense, Cirsium spinosum, Cirsium maritimum, C. irimtiense, C. yakushimense, etc. Among these plants from which pectinarigenin and/or its glycosides are derived, Cirsium brevicaule is a preferred embodiment.

In addition, in one embodiment of the SGLT inhibitor of the present disclosure, pectinarigenin and/or its glycosides may be purified, or may be a crude product, or may be in the form of a plant extract containing pectinarigenin and/or its glycosides, or may be a product obtained by simply processing a plant containing pectinarigenin and/or its glycosides by steaming, drying, roasting, shredding, crushing, or the like.

A plant extract containing pectinarigenin and/or its glycosides can be obtained, for example, by extracting the above-mentioned Cirsium plant of the Asteraceae family.

The parts of the Cirsium genus plant of the Asteraceae family that can be extracted include, for example, leaves, stems, roots, rhizomes, fruits, seeds, seed coats, flowers, etc., but in a preferred embodiment, the leaves are used. The parts of the Cirsium genus plant of the Asteraceae family that can be extracted may be subjected to pretreatment such as steaming, drying, roasting, shredding, or crushing, as necessary.

The extraction process may be any common extraction method used in the production of plant extracts, such as solvent extraction, supercritical extraction, steam distillation, etc. Among these, a preferred embodiment is solvent extraction.

The extraction solvent used in the solvent extraction process is not particularly limited as long as it is a solvent capable of extracting pectolinarigenin and/or its glycosides, and examples thereof include water; alcohols such as methanol, ethanol, propanol, and butanol; polyhydric alcohols such as ethylene glycol, propylene glycol, butylene glycol, and glycerin; ketones such as acetone and methyl ethyl ketone, esters such as methyl acetate and ethyl acetate; ethers such as tetrahydrofuran and diethyl ether; halogenated hydrocarbons such as dichloromethane, dichloroethane, and chloroform; aliphatic hydrocarbons such as hexane and pentane; aromatic hydrocarbons such as toluene; polyethers such as polyethylene glycol; organic solvents such as pyridines, and mixtures of these organic solvents and water. These extraction solvents may be used alone or in combination of two or more. Among these extraction solvents, a preferred embodiment is ethanol, or a mixture of ethanol and water.

Solvent extraction can be carried out by immersing or refluxing the part to be extracted in the extraction solvent, and a person skilled in the art can appropriately set the amount of extraction solvent used, the temperature during the extraction process, the extraction process time, etc.

By removing the residue after the solvent extraction process, a plant extract containing pectolinarigenin and/or its glycosides can be obtained. The residue can be removed using, for example, suction filtration, a filter press, a cylinder press, a decanter, a centrifuge, a filter centrifuge, etc.

If necessary, the obtained extract may be subjected to further concentration, washing, and purification processes such as concentration, drying, washing with water, washing with organic solvents, desalting, activated carbon treatment, recrystallization, distribution purification, column chromatography, and vacuum distillation to remove impurities. These processes may be performed by repeating the same process multiple times, or by combining two or more different processes. Furthermore, the obtained extract may be further subjected to processes such as powdering, granulation, and emulsification to adjust it to the desired shape.

The content of pectolinarigenin and/or its glycoside in the SGLT inhibitor of the present disclosure is not particularly limited, as long as it is an effective amount capable of inhibiting SGLT in the body, and can be set appropriately depending on the application, dosage form, administration form, etc.

[Other added ingredients]
In one embodiment of the SGLT inhibitor of the present disclosure, in addition to pectinarigenin and/or its glycoside, other additive components may be contained depending on the dosage form, formulation, etc. The type and content of the additive components are appropriately set depending on the dosage form, formulation, etc.

[Dosage form/formulation]
The dosage form of the SGLT inhibitor of the present disclosure is not particularly limited and may be any of solid, semi-solid, or liquid, and may be appropriately selected depending on the formulation form, administration method, etc. of the SGLT inhibitor.

The method of application of the SGLT inhibitor of the present disclosure is not particularly limited, and may be any of oral ingestion, oral administration, intravascular (intra-arterial or intravenous) administration, transdermal administration, enteral administration, pulmonary administration, and intranasal administration, but preferred embodiments include oral ingestion or oral administration.

The formulation of the SGLT inhibitor disclosed herein may be in the form of a food or drink, or a pharmaceutical.

When the SGLT inhibitor of the present disclosure is formulated into a food or beverage, the active ingredient may be prepared as is or in combination with other food materials or additives into the desired form. Examples of such food or beverage include general food or beverage products, as well as foods for specified health uses, foods with nutritional functions, and foods with functional claims. The form of these food or beverage products is not particularly limited, but specific examples include supplements such as capsules (soft capsules, hard capsules), tablets, granules, powders, jellies, and liposome preparations; beverages such as nutritional drinks, fruit juice drinks, carbonated drinks, and lactic acid drinks; and luxury items such as dumplings, ice cream, sherbet, gummies, and candies. Of these food or beverage products, supplements and beverages are preferred.

When the SGLT inhibitor of the present disclosure is formulated into a pharmaceutical formulation, the active ingredient may be prepared into the desired form either as is, in combination with other additive ingredients, or in combination with a delivery technique appropriate for the dosage form. Specific examples of such pharmaceuticals include oral medicines such as drinks, capsules (soft capsules, hard capsules), tablets, granules, powders, jellies, and syrups; liposome preparations, injections, enteral preparations, drips, nasal drops, injections, infusions, suppositories, and the like. Among these pharmaceuticals, oral medicines are preferred.

When the SGLT inhibitor of the present disclosure is in the form of a food or beverage or an oral pharmaceutical preparation, the content of the active ingredient pectolinarigenin and/or its glycoside is not particularly limited, as long as it is an effective amount capable of inhibiting SGLT in the body, and may be set appropriately depending on the dosage form and formulation, and may be, for example, 0.01 to 100% by mass, preferably 0.1 to 50% by mass, and more preferably 1 to 30% by mass.

[Application]
The SGLT inhibitor of the present disclosure is used for inhibiting SGLT in vivo. The SGLT subtypes to be inhibited by the SGLT inhibitor of the present disclosure may be any of SGLT1, SGLT2, and SGLT3, preferably SGLT1 and SGLT2, more preferably SGLT1.

For example, since the inhibition of SGLT1 can inhibit or delay the absorption of glucose in the body, in one embodiment of the SGLT inhibitor disclosed herein, the inhibition of SGLT1 can be used to prevent or treat diseases or symptoms caused in part by hyperglycemia.

Diseases and symptoms caused in part by hyperglycemia include, for example, diabetes (including both type 1 and type 2), impaired glucose tolerance, impaired fasting glucose, diabetic complications (e.g., diabetic retinopathy, diabetic nephropathy, diabetic neuropathy), hyperinsulinemia, hyperlipidemia, dyslipidemia, atherosclerosis, hyperuricemia, gout, and obesity (e.g., obesity due to metabolic syndrome). Among these, the SGLT inhibitor of the present disclosure is suitable for preventing or treating diabetes and diabetic complications, particularly type 2 diabetes.

The dosage of the SGLT inhibitor of the present disclosure is not particularly limited, and an effective amount capable of inhibiting SGLT in the body may be appropriately set depending on the formulation form, use, recipient, expected effect, etc. For example, when the SGLT inhibitor of the present disclosure is orally ingested or administered, the dosage may be about 0.01 to 3,000 mg, and preferably about 0.1 to 1,000 mg, of pectolinarigenin per day for an adult. The SGLT inhibitor of the present disclosure may be ingested or administered once or in several divided doses so that the daily amount falls within the aforementioned range.

The configurations and combinations thereof in each embodiment are merely examples, and additions, omissions, substitutions, and other modifications of the configurations are possible as appropriate without departing from the spirit of this disclosure. Each aspect disclosed in this specification may be combined with any other feature disclosed in this specification.

The following examples are provided for more detailed explanation, but the present invention is not limited to the embodiments, but is limited only by the scope of the claims.

Test Example 1: Evaluation of SGLT1 inhibitory activity of pectolinarigenin
A test solution was prepared by adding 10 μM pectolinarigenin or 10 μM chlorogenic acid to a KRH (Krebs Ringer Hepes) buffer solution containing 2 mM 2DG (2-deoxyglucose).

Human SGLT1 highly expressing CHO cells (recombinant cells in which human SGLT1 expression vector was introduced into CHO cells) were seeded in each well of a 24-well plate at 2.0 × 10 ⁵ cells/well using Ham's/F-12 medium containing 2 mg/mL G418, and cultured overnight at 37 ° C. Then, the culture supernatant of each well was removed, and 0.5 ml of the test solution was added to each well and incubated at 37 ° C. for 10 minutes. Next, the cells in the well were collected and washed, and then 0.1 M aqueous sodium hydroxide solution was added to dissolve the cells, and the cells were neutralized with 0.1 M hydrochloric acid. The 2DG in the obtained cell lysate was measured using a Glucose Uptake-Glo Assay kit (Promega). In addition, a test solution containing no pectolinarigenin or chlorogenic acid was used as a control, and the test was performed in the same manner. The ratio of the amount of 2DG in the cell lysate in each test solution to the amount of 2DG in the cell lysate in the control was calculated and used as the 2DG uptake rate.

The results are shown in Figure 1. As a result, it was confirmed that pectolinarigenin was able to reduce the amount of 2DG taken up more than chlorogenic acid, which is known to have SGLT1 inhibitory activity, and has excellent SGLT1 inhibitory activity.

Test Example 2: Evaluation of the relationship between pectinarigenin concentration and SGLT1 inhibitory activity
The 2DG uptake rate was determined in the same manner as in Test Example 1 above, using test solutions prepared by adding pectolinarigenin to 1.0, 2.5, 5.0, 10.0, and 25.0 μM of KRH (Krebs Ringer Hepes) buffer containing 2 mM 2DG.

The results are shown in Figure 2. As a result, it was confirmed that pectolinarigenin was able to reduce the amount of 2DG uptake in a concentration-dependent manner and inhibit SGLT1 activity in a concentration-dependent manner.

Test Example 3: Evaluation of SGLT1 inhibitory activity of pectolinarigenin glycoside
The 2DG uptake rate was determined in the same manner as in Test Example 1 above, using a test solution prepared by adding 5 μM pectolinarigenin or 5 μM pectolinarin to a KRH (Krebs Ringer Hepes) buffer solution containing 2 mM 2DG.

The results are shown in Figure 3. As a result, a decrease in the amount of 2DG taken up was observed even with pectolinarin compared to the control, confirming that not only pectolinarigenin but also its glycosides have SGLT1 inhibitory activity.

Claims (2)

A sodium-dependent glucose cotransporter inhibitor containing pectolinarigenin and/or its glycoside as the active ingredient. The sodium-dependent glucose cotransporter inhibitor according to claim 1 , which is used to inhibit sodium-dependent glucose cotransporter 1.