WO2020026570A1 - Glucose consumption promoter and glycolysis promoter - Google Patents

Abstract

Provided is a new drug capable of promoting the consumption of glucose. The glucose consumption promoter of the present invention is characterized by including at least one selected from the group consisting of 2-amino-1-cyclohexylethanol, 1-amino-2-propanol, 1,3-bis[tris(hydroxymethyl)methylamino]propane, N-cyclohexylethanolamine, diethanolamine, diethylamine, dipropylamine, morpholine, propylamine, triethanolamine, triethylamine, and trishydroxymethylaminomethane.

Description

Glucose consumption promoter and glycolytic promoter

The present invention relates to a glucose consumption promoter and a glycolysis promoter.

Diabetes is a disease in which the concentration of glucose (blood sugar level) in the blood continues to be high due to insufficient working of insulin. It is known that over a prolonged period of diabetes, high concentrations of glucose cause vascular damage, causing complications such as heart disease, blindness, renal failure, and amputation of the feet.

As a method of treating diabetes, for example, in the case of type 1 diabetes, supplementation of insulin from outside the body by insulin injection, and in the case of type 2 diabetes, in addition to diet therapy and exercise therapy, oral hypoglycemic drugs and insulin injection, etc. A method of controlling blood sugar level by drug therapy is known (Patent Document 1). However, there is a need for a new method capable of controlling the glucose concentration.

JP-A-2005-205925

Therefore, an object of the present invention is to provide a new drug capable of promoting consumption of glucose.

In order to achieve the above object, the glucose consumption promoter and the glycolysis promoter of the present invention include 2-amino-1-cyclohexylethanol, 1-amino-2-propanol, 1,3-bis [tris (hydroxymethyl ) Methylamino] propane, N-cyclohexylethanolamine, diethanolamine, diethylamine, dipropylamine, morpholine, propylamine, triethanolamine, triethylamine, trishydroxymethylaminomethane. Features.

医 薬 A pharmaceutical composition for use in the treatment of diabetes of the present invention is characterized by containing the glucose consumption promoter or glycolysis promoter of the present invention.

グ ル コ ー ス The glucose consumption promoting method of the present invention is characterized by including the step of administering the glucose consumption promoting agent of the present invention or the pharmaceutical composition of the present invention. Further, the glycolysis promoting method of the present invention includes a step of administering the glycolysis promoting agent of the present invention or the pharmaceutical composition of the present invention.

According to the present invention, a new drug capable of promoting glucose consumption can be provided.

FIG. 1 is a graph showing the relative value of glucose consumption of fibroblasts to which each drug was added in Example 1. FIG. 2 is a graph showing the relative values of glucose consumption of fibroblasts to which the respective drugs of the comparative example were added in Example 1. FIG. 3 is a graph showing relative values of glucose consumption of fibroblasts to which each drug was added in Example 2. FIG. 4 is a graph showing the relative values of glucose consumption of fibroblasts to which each drug was added in Example 3. FIG. 5 is a graph showing the relative value of glucose consumption of fibroblasts to which each drug was added in Example 4. FIG. 6 is a graph showing the relative value of glucose consumption of liver cancer cells to which each drug was added in Example 5. FIG. 7 is a graph showing the relative values of the lactate concentrations of fibroblasts and liver cancer cells to which each drug was added in Example 6. FIG. 8 is a graph showing the blood glucose level of the mice to which the drug was administered in Example 7. FIG. 9 is a graph showing the calculated value of AUC of the mouse to which the drug was administered in Example 7. FIG. 10 is a graph showing the lactic acid concentration in the culture solution of each cell to which the drug was administered in Example 8. FIG. 11 is a graph showing the lactic acid concentration in the culture solution of each cell to which the drug was administered in Example 9.

The glucose consumption promoter and glycolysis promoter of the present invention include, for example, 2-amino-1-cyclohexylethanol, 1-amino-2-propanol, 1,3-bis [tris (hydroxymethyl) methylamino] propane, It contains at least one selected from the group consisting of N-cyclohexylethanolamine, diethanolamine, diethylamine, dipropylamine, morpholine, propylamine, triethanolamine, triethylamine and trishydroxymethylaminomethane as a main component.

グ ル コ ー ス The glucose consumption promoter and glycolysis promoter of the present invention further include, for example, an oral additive.

用語 The terms used in the present specification can be used in the meaning usually used in the technical field unless otherwise specified.

Hereinafter, the present invention will be described in detail.

(Glucose consumption promoter and glycolytic promoter)
As described above, the glucose consumption promoter and the glycolysis promoter of the present invention include a compound represented by the following chemical formula (1), a tautomer and a stereoisomer thereof, and a salt thereof (hereinafter, referred to as “the present invention”). (Also referred to as “drug in the invention”). In the glucose consumption promoter and the glycolysis promoter of the present invention, other configurations and conditions are not particularly limited.

In the chemical formula (1), R ¹ is a hydrogen atom or a hydroxy group. Further, as described later, R ¹ and R ⁶ may be integrally formed to form a ring structure.

In the above chemical formula (1), R ² is a substituent containing a hydrogen atom, a linear or branched alkyl group, an aryl group, a cycloalkyl group or a hetero ring. The number of carbon atoms of the linear or branched alkyl group is not particularly limited, and may be, for example, 1 to 40, 1 to 32, 1 to 24, 1 to 18, 1 to 12, 1 to 6, or 1 to 2 (non- And 2 or more in the case of a saturated hydrocarbon group). The alkyl group is specifically, for example, a methyl group, an ethyl group, an n-propyl group, an isopropyl group, an n-butyl group, an isobutyl group, a sec-butyl group and a tert-butyl group, a pentyl group, a hexyl group, Heptyl, octyl, nonyl, decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, heptadecyl, octadecyl, nonadecyl, icosyl and the like. The same applies to a group derived from an alkyl group (hydroxyalkyl group). The aryl group includes, for example, a monocyclic aromatic hydrocarbon group and a polycyclic aromatic hydrocarbon group. Examples of the monocyclic aromatic hydrocarbon group include phenyl. Examples of the polycyclic aromatic hydrocarbon group include 1-naphthyl, 2-naphthyl, 1-anthryl, 2-anthryl, 9-anthryl, 1-phenanthryl, 2-phenanthryl, and 3-phenanthryl. Group, 4-phenanthryl group, 9-phenanthryl group and the like. Examples of the cycloalkyl group include a cyclopropyl group, a cyclobutyl group, a cyclopentyl group, a cyclohexyl group, a cycloheptyl group, and a cyclooctyl group. In the hetero ring in the substituent containing a hetero ring, the number of atoms constituting the ring is not particularly limited and is, for example, 3, 4, 5, 6, 7, 8, 9, and 10. The hetero atom in the substituent containing a hetero ring is, for example, at least one selected from the group consisting of O, S, N, and NH. Specific examples of the substituent containing a hetero ring include a group represented by the following chemical formula (R2). In the following chemical formula (R2), the atoms constituting the ring may be CH instead of N, O may be substituted for S, and S may be substituted for O.

場合 When there is an isomer in the substituent or the like, any isomer may be used unless otherwise specified.

One or more of the hydrogen atoms bonded to the carbon atoms of the substituents including the alkyl group, the aryl group, the cycloalkyl group and the hetero ring may be substituted with a further substituent. The further substituent is not particularly limited and includes, for example, carboxy, halogen, alkyl halide (eg, CF ₃ , CH ₂ CF ₃ , CH ₂ CCl ₃ ), nitro, nitroso, cyano, alkyl (eg, methyl, Ethyl, isopropyl, tert-butyl), alkenyl (eg, vinyl), alkynyl (eg, ethynyl), cycloalkyl (eg, cyclopropyl, adamantyl), cycloalkylalkyl (eg, cyclohexylmethyl, adamantylmethyl), cycloalkenyl ( Examples: cyclopropenyl), aryl (eg, phenyl, naphthyl), arylalkyl (eg, benzyl, phenethyl), heteroaryl (eg, pyridyl, furyl), heteroarylalkyl (eg, pyridylmethyl), heterocyclyl (eg, piperidyl) ), Heterosik Rylalkyl (eg, morpholylmethyl), alkoxy (eg, methoxy, ethoxy, propoxy, butoxy), perfluoroalkyl (eg, CF ₃ ), halogenated alkoxy (eg, OCF ₃ ), acyl, alkenyloxy (eg, vinyloxy, allyloxy) , Aryloxy (eg, phenyloxy), alkyloxycarbonyl (eg, methoxycarbonyl, ethoxycarbonyl, tert-butoxycarbonyl), arylalkyloxy (eg, benzyloxy), amino [alkylamino (eg, methylamino, ethylamino) , Dimethylamino), acylamino (eg, acetylamino, benzoylamino), arylalkylamino (eg, benzylamino, tritylamino), hydroxyamino], alkylaminoalkyl (eg, diethyl Aminomethyl), sulfamoyl, oxo and the like.

In the chemical formula (1), R ³ , R ⁴ , and R ⁵ are each a hydrogen atom, a linear or branched alkyl group, or a linear or branched hydroxyalkyl group. The linear or branched alkyl group and the linear or branched hydroxyalkyl group are, for example, as described above. R ³ , R ⁴ , and R ⁵ may each be the same or different. Further, as described later, R ⁵ and R ⁶ may be integrally formed to form a ring structure. One or more hydrogen atoms bonded to carbon atoms of the alkyl group and the hydroxyalkyl group may be substituted with a further substituent. The further substituent is, for example, as described above.

In the chemical formula (1), R ⁶ represents a hydrogen atom, a straight-chain or branched alkyl group, a straight-chain or branched hydroxyalkyl group, a cycloalkyl group, or a substituent containing a hetero atom, And may or may not include a cyclic structure. The linear or branched alkyl group, the linear or branched hydroxyalkyl group, and the cycloalkyl group are, for example, as described above. The carbon number of the substituent containing a hetero atom is not particularly limited, and may be, for example, 1 to 40, 1 to 32, 1 to 24, 1 to 18, 1 to 12, 1 to 6, or 1 to 2. Is also good. The hetero atom in the substituent containing a hetero atom is, for example, at least one selected from the group consisting of O, S, N, and NH. Specific examples of the substituent containing a heterocyclic ring include groups represented by the following chemical formulas (R6-1) and (R6-2). In the following chemical formulas (R6-1) and (R6-2), m, n, o, p, q, r, and s are each a positive integer and are not particularly limited, and for example, 1 to 10, 1 ５5, 1 ～ 3. Specific examples of the substituent containing a hetero ring include groups represented by the following chemical formulas (R6-1-2) and (R6-2-2). One or more of the hydrogen atoms bonded to the carbon atoms of the alkyl, hydroxyalkyl, cycloalkyl, and substituents including heteroatoms may be substituted with additional substituents. The further substituent is, for example, as described above.

In the chemical formula (1), R ¹ and R ⁶ may be combined to form a cyclic structure. The number of carbon atoms in the cyclic structure is not particularly limited, and may be, for example, 1 to 40, 1 to 32, 1 to 24, 1 to 18, 1 to 12, 1 to 6. The cyclic structure may or may not have a hetero atom, for example. The hetero atom is, for example, as described above. Specific examples of the cyclic structure include a structure represented by the following chemical formula (R1R6). In the following chemical formula (R1R6), R ² , R ³ , R ⁴ , and R ⁵ are, for example, as described above. One or more of the hydrogen atoms bonded to the carbon atoms of the cyclic structure may be substituted with a further substituent. The further substituent is, for example, as described above.

In the chemical formula (1), R ⁵ and R ⁶ may be combined to form a cyclic structure. The number of carbon atoms in the cyclic structure is not particularly limited, and may be, for example, 1 to 40, 1 to 32, 1 to 24, 1 to 18, 1 to 12, 1 to 6. The cyclic structure may or may not have a hetero atom, for example. The hetero atom is, for example, as described above. Specific examples of the cyclic structure include a structure represented by the following chemical formula (R5R6). In the following chemical formula (R5R6), R ¹ , R ² , R ³ , and R ⁴ are, for example, as described above. In the following chemical formula (R5R6), t is a positive integer and is not particularly limited, and is, for example, 1 to 10, 1 to 5, 1 to 3, or 2. Specific examples of the cyclic structure include a structure represented by the following chemical formula (R5R6-2). In the following chemical formula (R5R6-2), R ¹ , R ² , R ³ , and R ⁴ are, for example, as described above. One or more of the hydrogen atoms bonded to the carbon atoms of the cyclic structure may be substituted with a further substituent. The further substituent is, for example, as described above.

The compound represented by the chemical formula (1) is specifically, for example, 2-amino-1-cyclohexylethanol, 2-aminoethanol, 1-amino-2-propanol, 2-amino-1-phenylethanol, 1,3-bis [tris (hydroxymethyl) methylamino] propane, N-cyclohexylethanolamine, diethanolamine, diethylamine, dipropylamine, HEPES sodium salt, metoprolol tartrate, morpholine, octopamine, propylamine, triethanolamine, triethylamine , Timolol maleate, trishydroxymethylaminomethane and the like.

In Table 1 below, the structure of the drug of the present invention is shown by a combination of R ¹ , R ² , R ³ , R ⁴ , R ⁵ and R ^{6 in} the chemical formula (1). In Table 1, “chemical formula R6-1-2” and “chemical formula R6-2-2” are respectively represented by the chemical formula (R6-1-2) and the chemical formula (R6-2-2). Group. “(R5R6-2)” indicates that in the chemical formula (1), R ⁵ and R ⁶ together form a cyclic structure represented by the chemical formula (R5R6-2). . “(R1R6)” indicates that, in the chemical formula (1), R ¹ and R ⁶ are united to form a cyclic structure represented by the chemical formula (R1R6).

Each of the above-mentioned drugs is a known drug. In contrast, the present inventors have reported that these drugs, which are compounds represented by the chemical formula (1), promote the consumption of glucose and the glycolytic system, although the mechanism is unknown. The present inventors have obtained new findings and have found the present invention.

In the present invention, “consumption of glucose” may be, for example, promotion of a decrease in glucose concentration or suppression of an increase in glucose concentration. Glucose consumption can be measured, for example, by the method described in Examples described later. "Glucose consumption" may be, for example, the consumption of glucose by cells. In this case, “consumption of glucose” can also be referred to as, for example, “uptake of glucose into cells”. For this reason, the glucose consumption promoter of the present invention can also be referred to as, for example, a glucose uptake promoter into cells. In addition, it is known that the uptake of glucose is via, for example, a glucose transporter present in a cell membrane. From this, the glucose consumption promoter of the present invention can be said to be, for example, an activator of a signaling cascade via a glucose transporter. However, the present invention is not limited to this.

に お い て In the present invention, the “glycolysis system” is, for example, a metabolic system starting from glucose. Lactic acid is an example of the glycolytic metabolite. Therefore, for example, by measuring the lactic acid concentration as described later, it is possible to confirm the promotion of the glycolytic system. The glycolysis system is also referred to as, for example, an anaerobic glycolysis system.

According to the glucose consumption promoter of the present invention, glucose consumption can be promoted as described above. Further, according to the glycolytic accelerator of the present invention, as described above, the glycolytic accelerator can be promoted. Therefore, the glucose consumption promoter of the present invention and the glycolysis promoter of the present invention can be used, for example, as a pharmaceutical composition for use in treating a disease caused by glucose concentration in a living body. Examples of the disease include diabetes. In the present invention, “treatment” includes, for example, prevention of the disease, improvement of the disease, and improvement of prognosis of the disease, and may be any.

According to the glucose consumption promoting agent of the present invention, for example, it is possible to promote the consumption of glucose and the glycolysis system without suppressing the metabolism of lactic acid generated by the consumption of glucose.

グ ル コ ー ス The glucose consumption promoting agent of the present invention may contain, as an active ingredient, the above-mentioned agent of the present invention, for example, only one kind or two or more kinds in combination, and the number is not particularly limited. The glucose consumption promoting agent of the present invention contains, for example, the agent of the present invention as a main component.

The glucose consumption promoter of the present invention may be used, for example, in vivo , or may be used in vitro . The glucose consumption promoting agent of the present invention can be used, for example, as a research reagent, and as described above, can also be used as a pharmaceutical.

The administration target of the glucose consumption promoting agent of the present invention is not particularly limited. When the glucose consumption promoting agent of the present invention is used in vivo , the subject to which the glucose consumption promoting agent of the present invention is administered is not particularly limited, and examples thereof include humans and non-human animals other than humans. Examples of the non-human mammal include non-human animals such as mice, rats, rabbits, dogs, sheep, horses, cats, goats, monkeys, and guinea pigs. When the glucose consumption promoter of the present invention is used in vitro , the administration subject includes, for example, cells, tissues, organs and the like. The cells may be, for example, cells collected from a living body or cultured cells. The cells are not particularly limited, and include, for example, fibroblasts, hepatocytes, adipocytes and the like.

The use conditions (hereinafter, also referred to as “administration conditions”) of the glucose consumption enhancer of the present invention are not particularly limited, and for example, the administration form, administration timing, dosage amount, etc. are appropriately determined according to the type of administration subject and the like. Can be set. Examples of the administration form include oral administration, intraperitoneal administration, and subcutaneous administration when the glucose consumption promoter of the present invention is used in vivo .

剤 The dosage form of the glucose consumption promoter of the present invention is not particularly limited, and can be appropriately determined depending on, for example, the administration form. For example, in the case of oral administration, the dosage form is a capsule, an extract, an elixir, a granule, a pill, a suspension, a fine granule, a powder, an alcoholic beverage, a tablet, a syrup, an infusion / decoction, and a tincture. Agents, aromatic agents, limonade agents, liquid extract agents and the like.

The glucose consumption promoting agent of the present invention may, for example, if necessary, contain an additive.When the glucose consumption promoting agent of the present invention is used as a medicine, the additive is a pharmaceutically acceptable additive. preferable. Examples of the additives include excipients, stabilizers, preservatives, buffers, flavoring agents, suspending agents, emulsifiers, flavoring agents, solubilizing agents, coloring agents, thickeners, and the like. Examples of the additives include oral additives. Oral additives include, for example, caries preventives, intestinal medicines, corrigents and the like. In the present invention, the amount of the additive is not particularly limited as long as it does not hinder the function of the glucose consumption promoter.

The administration conditions of the glucose consumption enhancer of the present invention are not particularly limited, and for example, the administration timing, administration period, dosage, and the like can be appropriately set according to the type, gender, age, administration site, and the like of the administration subject. .

As a specific example, when the glucose consumption enhancer of the present invention is orally administered to a human, the total daily dose is, for example, 100 to 5000 mg and 500 to 2500 mg. The number of administrations per day is, for example, 1 to 5 times and 2 to 3 times.

(Pharmaceutical composition)
As described above, the pharmaceutical composition for use in the treatment of diabetes of the present invention is characterized by containing the glucose consumption promoter of the present invention or the glycolysis promoter of the present invention. The pharmaceutical composition of the present invention is characterized by containing the glucose consumption promoter of the present invention or the glycolysis promoter of the present invention, and other configurations and conditions are not particularly limited. The description of the glucose consumption promoter of the present invention and the glycolysis promoter of the present invention can be referred to for the pharmaceutical composition of the present invention. According to the pharmaceutical composition of the present invention, for example, diabetes can be treated.

(Method of promoting glucose consumption and method of promoting glycolysis)
The glucose consumption promoting method of the present invention includes a step of administering the glucose consumption promoting agent of the present invention to an administration subject. Further, the glycolysis promoting method of the present invention includes a step of administering the glycolysis promoting agent of the present invention to a subject to be administered. The present invention is characterized in that it comprises a step of administering the glucose consumption promoter or glycolysis promoter of the present invention, and other steps and conditions are not particularly limited. The glucose consumption promoter or glycolysis promoter of the present invention is as described above. The administration conditions and the like of the glucose consumption promoter and the glycolytic accelerator of the present invention are not particularly limited, and are, for example, the same as those described for the glucose consumption promoter of the present invention.

(Use of drugs)
The present invention is the use of the medicament of the present invention for use in promoting glucose consumption and glycolysis, and the use of the aforementioned medicament for use in treating diabetes. The present invention relates to the use of said medicament for the production of a glucose consumption promoter and a glycolytic accelerator, and the use of said medicament for the production of a pharmaceutical composition for use in the treatment of diabetes. is there. In the present invention, for example, the description of the glucose consumption promoting agent and the glycolytic accelerator, the pharmaceutical composition, and the glucose consumption promoting method and the glycolytic promoting method of the present invention can be cited.

Next, embodiments of the present invention will be described. However, the present invention is not limited by the following examples. Commercial reagents were used based on those protocols unless otherwise indicated.

[Example 1]
It was confirmed that the agent of the present invention exerts a glucose consumption promoting effect on fibroblasts.

As drugs, 2-Amino-1-cyclohexylethanol (manufactured by Matrix Biochemicals), 2-Aminoethanol (manufactured by Tokyo Chemical Industry), 1-amino-2- Propanol (Amino-2-propanol) (manufactured by Wako Pure Chemical Industries), 2-Amino-1-phenylethanol (manufactured by Tokyo Chemical Industry Co., Ltd.), 1,3-bis [tris (hydroxy Methyl) methylamino] propane (1,3-Bis [tris (hydroxymethyl) -methylamino] propane) (manufactured by Tokyo Chemical Industry Co., Ltd.), N-cyclohexylethanolamine (manufactured by Tokyo Chemical Industry Co., Ltd.), diethanolamine ( Diethanolamine (manufactured by Wako Pure Chemical), diethylamine (manufactured by Wako Pure Chemical), dipropylamine (manufactured by Tokyo Chemical Industry), sodium salt of HEPES (HEPES sodium salt) ) (Manufactured by MP Biomedicals), metoprolol tartrate (manufactured by Tokyo Chemical Industry), morpholine (Morpholine) (manufactured by Wako Pure Chemical Industries), octopamine (manufactured by MP Biomedicals), propylamine (Propylamine) ) (Manufactured by Tokyo Chemical Industry Co., Ltd.), triethylamine (manufactured by Wako Pure Chemical Industries, Ltd.), timolol maleate (Timolol Maleate) (manufactured by Wako Pure Chemical Industries, Ltd.), and trishydroxymethylaminomethane (Tris) (Wako Pure Chemical Industries, Ltd.) Was used. The drug was dissolved in distilled water to prepare a sample.

Then, rat-derived fibroblasts (Py-3Y1-S2, subcultured strain) are seeded on a 24-well microplate at a concentration of ～ 2 × 10 ⁵ cells / mL / well until a monolayer is formed. Cultured. In the culture, DMEM (manufactured by Nissui Pharmaceutical Co., Ltd.) was used as a medium. Each of the samples was added to the culture solution so that the final concentration of the drug was 0.5 mg / mL, and the cells were further cultured for 12 to 24 hours. As a drug control, culturing was performed in the same manner except that distilled water containing no drug was added to the culture solution instead of the sample.

(4) Immediately after the addition of the drug, immediately after the start of the culture and after the culture, the glucose concentration of each culture solution was measured. The glucose concentration was measured using a glucose assay kit (Wako). Then, the glucose concentration after the culture was divided by the glucose concentration (5.6 mmol / L) immediately after the start of the culture to calculate glucose consumption. Further, the glucose consumption in the control was set as a reference value of 100, and the relative value of the glucose consumption in each sample was calculated.

The results are shown in FIG. FIG. 1 (A) is a graph showing relative values of glucose consumption of fibroblasts when 0.5 mg / mL of each of the above-mentioned drugs was added. In FIG. 1 (A), the vertical axis indicates a relative value of glucose consumption, and the horizontal axis indicates a drug. As shown in FIG. 1 (A), in each of the samples, the relative value of the glucose consumption increased to 100 or more and the glucose consumption increased due to the addition of each drug. Among them, 2-amino-1-cyclohexylethanol, 2-aminoethanol, 1-amino-2-propanol, 2-amino-1-phenylethanol, N-cyclohexylethanolamine, diethanolamine, diethylamine, dipropylamine, sodium salt of HEPES , Metoprolol tartrate, morpholine, propylamine, triethylamine, timolol maleate, and trishydroxymethylaminomethane significantly increased glucose consumption compared to controls (t-test, p <0.05 , Or p <0.1, which is represented by “**” or “*” in FIG. 1 (A)).

Further, as drugs, 2-amino-1-cyclohexylethanol, 2-aminoethanol, 1-amino-2-propanol, 2-amino-1-phenylethanol, 1,3-bis [tris (hydroxymethyl) methylamino] Propane, N-cyclohexylethanolamine, diethanolamine, diethylamine, dipropylamine, sodium salt of HEPES, metoprolol tartrate, morpholine, propylamine, triethanolamine (Triethanolamine) (manufactured by Tokyo Chemical Industry Co., Ltd.), triethylamine, timolol maleate, The effect of promoting glucose consumption was confirmed in the same manner as described above, except that trishydroxymethylaminomethane was used and the final concentration of each of the above-mentioned drugs was 1 mg / mL.

The results are shown in FIG. FIG. 1 (B) is a graph showing the relative value of glucose consumption when 1 mg / mL of each of the above-mentioned drugs was added. In FIG. 1 (B), the vertical axis indicates the relative value of glucose consumption, and the horizontal axis indicates the drug. As shown in FIG. 1 (B), in each of the samples, the relative value of the glucose consumption increased to 100 or more and the glucose consumption increased due to the addition of each of the agents. Among them, 2-amino-1-cyclohexylethanol, 2-aminoethanol, 2-amino-1-phenylethanol, 1,3-bis [tris (hydroxymethyl) methylamino] propane, N-cyclohexylethanolamine, diethanolamine, diethylamine , Dipropylamine, sodium salt of HEPES, metoprolol tartrate, morpholine, propylamine, triethylamine, timolol maleate, trishydroxymethylaminomethane significantly increased glucose consumption compared to control ( t-test, p <0.05, or p <0.1, represented by “**” or “*” in FIG. 1 (B)), it was confirmed that a strong glucose consumption promoting effect was exhibited.

Next, as a comparative example, the same conditions as above were used except that Tricine and Trimethylolpropane, which are drugs having no structure represented by the chemical formula (1), were used. The effect of promoting glucose consumption was confirmed.

The results are shown in FIG. FIG. 2 is a graph showing relative values of glucose consumption when each drug of the comparative example was added. (A) shows a case where 0.5 mg / mL of each drug of the comparative example was added. (B) shows the results obtained when 1 mg / mL of each drug of the comparative example was added. 2A and 2B, the vertical axis indicates the relative value of glucose consumption, and the horizontal axis indicates the drug. As shown in FIGS. 2 (A) and (B), the relative value of the glucose consumption was less than 100 by adding 0.5 mg / mL or 1 mg / mL of each drug of the comparative example to each sample. And the glucose consumption did not increase.

か ら From the results of FIG. 1 and FIG. 2, it was confirmed that the drug of the present invention has a structure represented by the chemical formula (1) and thus has a glucose consumption promoting effect.

[Example 2]
It was confirmed that the agent of the present invention exerts a glucose consumption promoting effect on fibroblasts in a dose-dependent manner.

前 記 The above samples of 2-amino-1-cyclohexylethanol and 2-amino-1-phenylethanol, which were confirmed to have a strong effect of promoting glucose consumption in Example 1, were prepared. The fibroblasts were cultured under the same conditions as in Example 1. Each sample was added to this culture solution such that the final concentration of the drug was 0.1, 0.25, and 0.5 mg / mL, respectively. Then, the relative value of the glucose consumption in each sample was calculated in the same manner as in Example 1.

These results are shown in FIG. FIG. 3 is a graph showing relative values of glucose consumption of fibroblasts to which the above-mentioned drugs were added. (A) shows the results of adding 2-amino-1-cyclohexylethanol, and (B) shows the results. And the results of adding 2-amino-1-phenylethanol. 3A and 3B, the vertical axis indicates the relative value of glucose consumption, and the horizontal axis indicates the concentration of each drug. As shown in FIG. 3 (A), in fibroblasts, addition of 0.1, 0.25, and 0.5 mg / mL of 2-amino-1-cyclohexylethanol resulted in about 120, about 130, And up to about 150, the dose-dependent increase in glucose consumption. Then, at all concentrations, the glucose concentration was significantly increased as compared to the control (t test, p <0.05, represented by “**” in FIGS. 3 (A) and (B)), It was confirmed that a strong glucose consumption promoting effect was exhibited. As shown in FIG. 3B, addition of 0.1, 0.25, and 0.5 mg / mL of 2-amino-1-phenylethanol resulted in about 130, about 140, and about 160, respectively. Up to a dose dependent increase in glucose consumption. Then, the glucose concentration was significantly increased as compared with the control, and it was confirmed that a strong glucose consumption promoting effect was exhibited.

As shown in each graph of FIG. 3, all of the administered drugs showed a glucose consumption promoting effect in a dose-dependent manner. In addition, it was found that even at a low dose, the effect of promoting glucose consumption was effectively exerted.

[Example 3]
It was confirmed that the agent of the present invention exerted a glucose consumption promoting effect on fibroblasts in which glucose consumption was suppressed.

First, in the same manner as in Example 1, samples of streptozotocin (STZ), alloxan (Alloxan), and nicotinamide (Nicotinamide) were prepared. The fibroblasts were cultured. Each of the samples was added to this culture solution so as to be 1 mg / mL. Streptozotocin, alloxan, and nicotinamide are all drugs known to exhibit symptoms of diabetes when administered to a living body. Then, the relative value of the glucose consumption was calculated in the same manner as in Example 1.

サ ン プ ル Next, each sample to which 2-amino-1-phenylethanol (2-A-1-P) was added in addition to streptozotocin, alloxan, and nicotinamide was prepared in the same manner. The concentration of 2-A-1-P was 1 mg / mL. Then, the relative value of the glucose consumption was calculated in the same manner.

The results are shown in FIG. FIG. 4 is a graph showing relative values of glucose consumption of fibroblasts to which each of the above-mentioned drugs was added. In FIG. 4, the vertical axis indicates the relative value of glucose consumption, and the horizontal axis indicates the drug. As shown in FIG. 4, by adding 1 mg / mL of streptozotocin, alloxan, and nicotinamide, the relative values of the glucose consumption became about 90, about 70, and about 85, respectively, and the glucose consumption was reduced. I was On the other hand, as a result of adding 2-A-1-P in addition to streptozotocin, alloxan, and nicotinamide, the relative value of the glucose consumption became 100 or more in all cases, and the glucose consumption recovered. Was. Thus, 2-amino-1-phenylethanol was confirmed to have an effect of promoting glucose consumption even on fibroblasts in which glucose consumption was suppressed.

[Example 4]
It was confirmed that the drug of the present invention further exerted a glucose consumption promoting effect on fibroblasts that promoted glucose consumption.

First, in the same manner as in Example 1, the samples of vanadium, V ₂ O ₅ , and concanavalin A (ConA) were prepared. The fibroblasts were cultured. Each sample was added to this culture solution so that vanadium was 1.0 mg / mL and ConA was 100 μg / ml. Vanadium, V ₂ O ₅ , and concanavalin A are all drugs known to show an insulin-like effect when administered to fibroblasts. Then, the relative value of the glucose consumption in each sample was calculated in the same manner as in Example 1.

Next, in addition to vanadium, V ₂ O ₅ , and concanavalin A, each sample to which 2-amino-1-phenylethanol (2-A-1-P) was further added was prepared in the same manner. The concentration of 2-A-1-P was 0.5 mg / mL, respectively. Then, the relative value of the glucose consumption was calculated in the same manner.

The result is shown in FIG. FIG. 5 is a graph showing the relative values of glucose consumption of fibroblasts to which the above-mentioned drugs were added. In FIG. 5, the vertical axis indicates the relative value of glucose consumption, and the horizontal axis indicates the drug. As shown in FIG. 5, the addition of vanadium, V ₂ O ₅ , and concanavalin A resulted in a relative value of the glucose consumption of about 110, about 115, and about 120, indicating an increase in glucose consumption. In contrast, as a result of adding 2-A-1-P in addition to vanadium, V ₂ O ₅ , and concanavalin A, the relative values of the glucose consumption were about 145, about 140, and about 140, respectively. It was 150 and the glucose consumption was further increasing. Thus, it was confirmed that 2-amino-1-phenylethanol further exerts a glucose consumption promoting effect on fibroblasts whose glucose consumption has been promoted by vanadium, V ₂ O ₅ , and concanavalin A. Was.

[Example 5]
It was confirmed that the agent of the present invention exerts a glucose consumption promoting effect on liver cancer cells.

First, in the same manner as in Example 1, the samples of 2-amino-1-cyclohexylethanol and 2-amino-1-phenylethanol were prepared. A rat-derived hepatoma cell (Ry121B, subcultured) was used in place of the rat-derived fibroblast (Py-3Y1-S2, subcultured) under the same conditions as in Example 1 above. Cultured. Each sample was added to this culture solution so as to be 1 mg / mL. As a control, culturing was performed in the same manner except that distilled water containing no drug was added to the culture solution instead of the sample. Then, the relative value of the glucose consumption in each sample was calculated in the same manner as in Example 1.

These results are shown in FIG. FIG. 6 is a graph showing the relative value of glucose consumption of liver cancer cells to which the above-mentioned drugs were added. In FIG. 6, the vertical axis indicates the relative value of glucose consumption, and the horizontal axis indicates the drug.

As shown in FIG. 6, even in hepatoma cells, by adding 2-amino-1-cyclohexylethanol and 2-amino-1-phenylethanol, the relative value of the glucose consumption compared to the control was about 145 and about 140, indicating that glucose consumption was significantly increased as compared to the control.

か ら From the results in FIG. 6, it was confirmed that the agent of the present invention also exerts a glucose consumption promoting effect on liver cancer cells.

[Example 6]
It was confirmed that the glucose consumption promoting effect of the agent of the present invention promotes glycolysis by cells.

First, in the same manner as in Example 1, the samples of 2-amino-1-cyclohexylethanol and 2-amino-1-phenylethanol were prepared. The fibroblasts and hepatoma cells were cultured under the same conditions as in Examples 1 and 5. Each of the samples was added to each of the culture solutions so as to be 1 mg / mL. As a control, culturing was performed in the same manner except that distilled water containing no drug was added to the culture solution instead of the sample.

(4) After the addition of the drug, the culture solution was diluted so that the weight ratio of culture solution to distilled water was 1:19, and the lactic acid concentration of the diluted culture solution was measured. The lactic acid concentration was measured using a lactic acid assay kit (product name: Lactate @ Assay @ Kit-WST, manufactured by Dojin Chemical Co., Ltd.). The relative value of the lactic acid concentration in each sample was calculated using the lactic acid concentration in the control as the reference value 100.

The results are shown in FIGS. 7 (A) and (B). FIG. 7 is a graph showing the relative values of the lactate concentration of each cell to which each of the above-mentioned drugs was added, (A) showing the result in fibroblasts, and (B) showing the result in liver cancer cells. . 7A and 7B, the vertical axis indicates the relative value of the lactic acid concentration, and the horizontal axis indicates the drug.

As shown in FIG. 7 (A), in fibroblasts, the addition of 2-amino-1-cyclohexylethanol and 2-amino-1-phenylethanol increased the lactic acid concentration to about 145 and about 185, respectively. I was In addition, as shown in FIG. 7 (B), the lactic acid concentration of the hepatoma cells was increased to about 140 and about 135 by adding 2-amino-1-cyclohexylethanol and 2-amino-1-phenylethanol, respectively. Was rising.

か ら From the results of FIGS. 7 (A) and (B), it was confirmed that the agent of the present invention increased the lactate concentration in fibroblasts and hepatoma cells and exhibited a glycolytic promoting effect.

[Example 7]
It was confirmed that the agent of the present invention reduced the blood glucose level of mice.

２－ 2-amino-1-phenylethanol was used as a drug. First, glucose (manufactured by Kanto Chemical Co., Ltd.) was dissolved in water for injection to a concentration of 200 mg / mL to prepare a glucose solution. Next, after dissolving the drug in the water for injection, the glucose solution is added to the solution so that the final concentrations of the drug and glucose are 2.5 mg / mL and 100 mg / mL, respectively. A sample was prepared. As a control of the drug, a 100 mg / mL glucose solution containing no drug was used instead of the sample.

6-week-old ICR male mice were purchased from Japan SLC, Inc., and were reared for about one week to confirm that there was no abnormality in their general condition. The conditions for the preliminary breeding were as follows: four mice were each housed in a polycarbonate cage, the room temperature was 23 ° C. ± 3 ° C., and the lighting time was 12 hours / day. Feed (solid feed for mice and rats, manufactured by Nippon Nosan Kogyo Co., Ltd.) and drinking water (tap water) were freely taken.

After the preliminary breeding, the mice were fasted for about 21 hours, and the body weight and the blood glucose level were measured. The test group and the control group were divided into a total of two groups so that blood glucose levels did not vary among the groups. The number of animals in each group was eight. Table 2 shows the weight (g) of the mice at the time of the grouping.

(4) After the grouping, the sample and the control were orally administered to the mice of the test group and the control group once, respectively, using a gastric tube so as to have a dose volume of 20 mL / kg. That is, the mice in the test group were administered with the drug and glucose at doses of 50 mg / kg and 2000 mg / kg, respectively, and the mice in the control group were administered glucose at a dose of 2000 mg / kg. Was administered. The blood glucose level was measured at 30 minutes, 60 minutes, 90 minutes, and 120 minutes, with the time of the administration being 0 minutes. Blood glucose was measured by using Accu-Chek Aviva (Roche Diagnostics Co., Ltd.), puncturing the tip of the tail with a syringe needle, collecting blood, and measuring the blood glucose of the collected blood.

比較 The test group and the control group were compared by a t-test for blood sugar levels at 0, 30, 60, 90 and 120 minutes. The significance level was 5% and 1%.

The results are shown in Table 3 and FIG. Table 3 is a table showing measured values (mg / dL) of blood glucose levels at a predetermined time after the administration in each individual. FIG. 8 shows blood glucose levels at a predetermined time after the administration in the test group and the control group. It is a graph which shows the average value of a value. In FIG. 8, the vertical axis indicates the blood glucose level (mg / dL), and the horizontal axis indicates the time (minute) after administration. In FIG. 8A, the average value was calculated based on the blood glucose levels obtained for all the individuals (No. 1 to No. 8) in the control group and the test group. On the other hand, as shown in Table 3, the blood glucose level of one individual (No. 8) in the test group after glucose administration was calculated for the other seven individuals (No. 1 to No. 7) in the test group. Abnormal values were indicated as compared to the values. For this reason, in FIG. 8B, the data of one individual (No. 8) in the test group showing the abnormal value is excluded, and the other seven individuals (No. 1 to No. 7) in the test group are excluded. The average value was calculated based on the blood glucose level obtained for (2).

As shown in FIGS. 8 (A) and 8 (B), the test group tended to have a lower blood glucose level than the control group at 30, 60, 90 and 120 minutes after administration. It was observed. In particular, at the time 30 minutes after administration, the test group had significantly lower blood glucose levels compared to the control group (P <0.05 and P <0.01).

Further, AUC (area @ under @ the @ curve) was calculated for the measured blood sugar level. The AUC is calculated by calculating an area surrounded by a straight line parallel to the time axis passing through the measured value at the time of administration and a measured value curve in a graph in which the vertical axis represents measured values and the horizontal axis represents time. Was. Then, the AUC was compared with the control group by the t-test in the same manner as in the above-mentioned comparison with respect to the blood glucose level.

The results are shown in Table 4 and FIG. Table 4 is a table showing the calculated value of AUC (mg / dL · h) in each individual, and FIG. 9 is a graph showing the average value of AUC in the test group and the control group. In FIG. 9, the vertical axis indicates AUC (mg / dL · h), and the horizontal axis indicates the control group and the test group. In FIG. 9A, the average value was calculated based on the calculated values of AUC obtained for all the individuals (No. 1 to No. 8) in the control group and the test group. On the other hand, as shown in Table 4, the calculated value (258) of the AUC of one individual (No. 8) in the test group was the same as that of the other seven individuals (No. 1 to No. 7) in the test group. Abnormal values were shown as compared to the calculated values. For this reason, in FIG. 9B, the data of one individual (No. 8) in the test group showing the abnormal value is excluded, and the other seven individuals (No. 1 to No. 7) in the test group are excluded. The average value was calculated on the basis of the calculated value of AUC obtained for ()).

示 す As shown in FIG. 9 (A), the test group tended to have a lower AUC value than the control group. Also, as shown in FIG. 9 (B), the test group had a significantly lower AUC value than the control group (P <0.05).

か ら From the results of FIGS. 8 and 9, it was confirmed that the agent of the present invention reduced the blood glucose level of mice.

Example 8
It was confirmed that the drug of the present invention exhibited a lactate synthesis promoting effect even after long-term culture.

First, in the same manner as in Example 1, the sample of 2-amino-1-cyclohexylethanol (2-amino-1-cyclohexylEOH) was prepared. In addition to the rat-derived fibroblasts (Py-3Y1-S2, subcultured), human-derived esophageal cancer cells (TE-13, subcultured) and African green monkey-derived kidney epithelial cells (VERO) , A subculture) and a human-derived hepatoma cell (HepG2, subculture) were cultured under the same conditions as in Example 1 above. Each of the samples was added to this culture solution to a concentration of 0.5 mg / mL. Thereafter, the cells were further cultured for 24-48 hours. The culture time after the addition of the sample was the same for each cell. Control 1 was cultured in the same manner except that distilled water without drug was added to the culture solution instead of the sample. As control 2, a biguanide (Biguanide) sample was prepared in the same manner as in Example 1 and cultured in the same manner except that the sample was added instead of the sample. Then, in the same manner as in Example 6, the lactic acid concentration in each sample was measured. Using the above four types of cells, a total of five experiments (once for Py-3Y1-S2, TE-13 and HepG2, and twice for VERO) were performed.

The results are shown in FIG. FIG. 10 is a graph showing the lactic acid concentration in the culture solution of each cell to which each of the above-mentioned drugs was added. The values in each graph show the average value of the results of the above five experiments. In FIG. 10, the vertical axis indicates the lactic acid concentration (mmol / L) in the culture solution, and the horizontal axis indicates the drug. As shown in FIG. 10, after adding 0.5 mg / mL of 2-amino-1-cyclohexylethanol and culturing for 24-48 hours, the lactic acid concentration was 9.35 mmol / L, and control 1 (Control) and increased (P <0.05). In control 2 (Biguanide), the addition of biguanide also increased the lactic acid concentration (P <0.01). Thus, it was confirmed that 2-amino-1-cyclohexylethanol increased the lactic acid concentration even after long-term culture, and exhibited a glycolytic promoting effect.

グ ル コ ー ス It is considered that most of the glucose in the culture solution has been changed to lactic acid after culturing for 24 to 48 hours. For example, since two molecules of lactic acid are synthesized from one molecule of glucose, if all of the glucose (5.6 mmol / L) immediately after the start of the culture is changed to lactic acid, the concentration of lactic acid in the culture solution is 11.2 mmol. / L. Then, as shown in FIG. 10, the lactic acid concentration in the culture solution when the culture was performed for 24 to 48 hours after the addition of each sample was about 8 to 10 mmol / L. From this, it can be said that, after the addition of each sample, about 70 to 90% of glucose, for example, was consumed and changed to lactic acid by the culture for 24 to 48 hours, respectively. In addition, it can be said that 2-amino-1-cyclohexylethanol exhibits a glucose consumption promoting effect even in the presence of a low concentration of glucose, for example. However, the present invention is not limited to this.

[Example 9]
It was confirmed that the drug of the present invention did not suppress lactate metabolism.

First, samples of 2-amino-1-cyclohexylethanol (2-amino-1-cyclohexylEOH) and 2-amino-1-phenylethanol (2-amino-1-phenylEOH) were prepared in the same manner as in Example 1. Prepared. In the same manner as in Example 8, each of the four types of cells was used and cultured under the same conditions as in Example 1. Each of the samples was added to this culture solution to a concentration of 0.5 mg / mL. Thereafter, the culturing time was further extended than in Example 8, and culturing was performed for 2 to 3 days (48 to 72 hours). Control 1 was cultured in the same manner except that distilled water without drug was added to the culture solution instead of the sample. Control 2 was similarly cultured except that a biguanide sample prepared in the same manner as in Example 1 was added to the culture solution instead of the sample. Then, in the same manner as in Example 6, the lactic acid concentration in each sample was measured. Using the above four types of cells, a total of four experiments were performed.

The results are shown in FIG. FIG. 11 is a graph showing the lactate concentration of each cell to which each of the above-mentioned drugs was added. The values in each graph indicate the average of the results of the four experiments. In FIG. 11, the ordinate indicates the lactic acid concentration (mmol / L) in the culture solution, and the abscissa indicates the drug. As shown in FIG. 11, after adding 0.5 mg / mL of 2-amino-1-cyclohexylethanol and 2-amino-1-phenylethanol and culturing for 2 to 3 days, the lactic acid concentration was respectively increased. Was about the same value as Control 1 (Control). On the other hand, in control 2 (Biguanide), the lactic acid concentration was about 3.5 times that of control 1 when culture was performed for 2 to 3 days after the addition of biguanide (P <0.05). . Here, it is known that lactic acid in a culture solution produced as a glycolytic metabolite is subsequently metabolized. This indicated that 2-amino-1-cyclohexylethanol and 2-amino-1-phenylethanol metabolized lactic acid to the same extent as control 1, and did not inhibit the metabolism of lactic acid. On the other hand, it was confirmed that biguanide suppressed the metabolism of lactic acid.

Although the present invention has been described with reference to the exemplary embodiments and examples, the present invention is not limited to the exemplary embodiments and examples. Various changes that can be understood by those skilled in the art can be made to the configuration and details of the present invention within the scope of the present invention.

This application is based on Japanese Patent Application No. 2018-143056 filed on Jul. 31, 2018, Japanese Patent Application No. 2018-181302 filed on Sep. 27, 2018, and International Application PCT / JP2019 / 003915. And all of its disclosures are incorporated herein.

As described above, according to the present invention, by containing at least one selected from the group consisting of the compound represented by the chemical formula (1), tautomers and stereoisomers thereof, and salts thereof , Can promote glucose consumption. And, since the drug can promote glucose consumption in this way, it can be used, for example, as a therapeutic agent for diabetes. Therefore, the present invention can be said to be extremely useful in the field of medicine and the like.

Claims (10)

1. 2-amino-1-cyclohexylethanol, 1-amino-2-propanol, 1,3-bis [tris (hydroxymethyl) methylamino] propane, N-cyclohexylethanolamine, diethanolamine, diethylamine, dipropylamine, morpholine, propyl A glucose consumption promoter comprising at least one selected from the group consisting of amines, triethanolamine, triethylamine, and trishydroxymethylaminomethane.
2. 2-amino-1-cyclohexylethanol, 1-amino-2-propanol, 1,3-bis [tris (hydroxymethyl) methylamino] propane, N-cyclohexylethanolamine, diethanolamine, diethylamine, dipropylamine, morpholine, propyl The glucose consumption enhancer according to claim 1, comprising at least one selected from the group consisting of amine, triethanolamine, triethylamine, and trishydroxymethylaminomethane as a main component.
3. The glucose consumption enhancer according to claim 1 or 2, further comprising an oral additive.
4. A pharmaceutical composition for use in the treatment of diabetes, comprising the glucose consumption enhancer according to any one of claims 1 to 3.
5. A method for promoting glucose consumption, comprising a step of administering the glucose consumption promoting agent according to any one of claims 1 to 3 or the pharmaceutical composition according to claim 4.
6. 2-amino-1-cyclohexylethanol, 1-amino-2-propanol, 1,3-bis [tris (hydroxymethyl) methylamino] propane, N-cyclohexylethanolamine, diethanolamine, diethylamine, dipropylamine, morpholine, propyl A glycolytic accelerator comprising at least one selected from the group consisting of amine, triethanolamine, triethylamine, and trishydroxymethylaminomethane.
7. 2-amino-1-cyclohexylethanol, 1-amino-2-propanol, 1,3-bis [tris (hydroxymethyl) methylamino] propane, N-cyclohexylethanolamine, diethanolamine, diethylamine, dipropylamine, morpholine, propyl The glycolytic accelerator according to claim 6, comprising at least one selected from the group consisting of amine, triethanolamine, triethylamine, and trishydroxymethylaminomethane as a main component.
8. The glycolytic accelerator according to claim 6 or 7, further comprising an oral additive.
9. A pharmaceutical composition for use in the treatment of diabetes, comprising the glycolytic accelerator according to any one of claims 6 to 8.
10. A method for promoting glycolysis, comprising a step of administering the glycolytic accelerator according to any one of claims 6 to 8 or the pharmaceutical composition according to claim 9.

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