JP2014148474A - Glp-1 secretion promoter - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a novel GLP-1 secretion promoter and use thereof.SOLUTION: There is provided a GLP-1 secretion promoter that comprises curcumins or curcumin derivatives (where, they have an ortho methoxyl phenolic group and a β-diketone structure), or salts thereof as an active ingredient.

Description

  The present invention relates to a GLP-1 secretion promoter and use thereof.

  GLP-1 (Glucagon-like peptide-1) is a peptide hormone (incretin) that is secreted from the gastrointestinal tract with food intake and acts on pancreatic β cells to promote insulin (blood sugar lowering hormone) secretion. Other actions of GPL-1 are known to promote pancreatic β-cell proliferation, suppress gastric excretion and gastric acid secretion, and suppress appetite and food intake. Therefore, promoting the secretion of GLP-1 and enhancing its effect are useful for the prevention and improvement of obesity and diabetes.

  Incretin includes GIP-1 and GIP (glucose-dependent insulinotropic polypeptide). For example, in type 2 diabetic patients, GIP-1 decreases the secretory effect of insulin secretion. It does not decline. Therefore, GLP-1 is important as a target for prevention and improvement of diabetes and the like. In fact, GLP-1-related drugs have been developed. Specifically, there are DPP-4 inhibitors and degradation-resistant GLP-1 derivative drugs that focus on the degradation of GLP-1 by the degradation enzyme (Dipeptidyl peptidase-4; DPP-4) present in the blood. It was developed (Non-Patent Document 1). However, the former has a problem of side effects due to inhibition of the degradation of the functional protein that should be originally degraded by DPP-4, and the latter has a problem that it cannot be administered orally because it is a peptide preparation.

  GLP-1 is not suitable for oral intake because it is a peptide hormone. Moreover, GLP-1 is very easily degraded, and the half-life in vivo is about several minutes. Therefore, it is desirable to promote endogenous GLP-1 secretion rather than external administration. In addition, as a substance which accelerates | stimulates the secretion of GLP-1, k-casein (patent document 1), soybean protein degradation product (patent document 2), Kiraya extract (patent document 3), and lysophosphatidylinositol (patent document 4). It has been reported.

International Publication No. 2007/037413 Pamphlet JP 2010-138-143 A JP 2012-131742 A JP 2012-144518 A

J. Clin. Endocrinol. Metab. 2006, 91: 4612-9, JAMA 2007, 50: 194-256

  Under the above background, an object of the present invention is to provide a novel GLP-1 secretion promoter and use thereof.

［３］糖尿病、肥満、食後高血糖又は神経変性疾患の予防又は改善、糖尿病に伴う神経障害の改善、グルカゴン分泌抑制、胃排泄や胃酸分泌の抑制、食欲や摂食の抑制、又は膵β細胞の増殖促進のために用いられる、［１］又は［２］に記載のGLP-1分泌促進剤。
［４］［１］又は［２］に記載のGLP-1分泌促進剤を含有する、糖尿病又は肥満の予防又は改善用組成物。
［５］医薬、医薬部外品又は食品である、［４］に記載の組成物。 In view of the above problems, the present inventor has repeatedly studied for the discovery of a GLP-1 secretion promoting substance. In particular, we focused on safety, and tried to identify substances with a high GLP-1 secretion-promoting action among naturally derived components. As a result, it was revealed that curcumin and its derivatives, which are components of turmeric (turmeric), exert a high GLP-1 secretion promoting action. In addition, structural features important for the action were found. Based on these results, the inventions listed below are provided.
[1] A GLP-1 secretion promoter comprising curcumin or a curcumin derivative (provided having an orthomethoxylphenol group and a β-diketone structure) or a salt thereof as an active ingredient.
[2] The GLP-1 secretion promoter according to [1], wherein the curcumin derivative is represented by the following chemical formula.

[3] Diabetes, obesity, postprandial hyperglycemia or neurodegenerative disease prevention or improvement, improvement of neuropathy associated with diabetes, suppression of glucagon secretion, suppression of gastric excretion and gastric acid secretion, suppression of appetite and feeding, or pancreatic β cells The GLP-1 secretion promoter according to [1] or [2], which is used for promoting the growth of.
[4] A composition for preventing or improving diabetes or obesity, comprising the GLP-1 secretion promoter according to [1] or [2].
[5] The composition according to [4], which is a medicine, quasi drug or food.

Structure of curcumin and curcumin derivatives. The graph which shows the result of an experiment. GLP-1 concentration in the culture supernatant 2 hours after administration of positive control (add glutamine to 10 mM as final concentration) or various curcumin derivatives at a final concentration of 25 μM to mouse gut-derived cells (Glutag) The result of having measured is shown. Different alphabetic characters on the bar graph indicate that they are significantly different from each other. The graph which shows the result of an experiment. In positive control (add glutamine to a final concentration of 10 mM) to mouse gut-derived cells (Glutag) or curcumin to a final concentration of 10 μM, 25 μM, 50 μM, and in the culture supernatant after 2 hours The result of having measured the GLP-1 density | concentration of is shown. Different alphabetic characters on the bar graph indicate that they are significantly different from each other.

  The first aspect of the present invention relates to a GLP-1 secretion promoter (hereinafter sometimes referred to as “the drug of the present invention” for convenience of explanation). As used herein, “GLP-1 secretion promotion” refers to promoting GLP-1 secretion by acting directly or indirectly on GLP-1 producing cells. Typically, when GLP-1 secretion is promoted, the amount of GLP-1 in the body increases, resulting in an increase in blood GLP-1 concentration or a decrease in blood GLP-1 concentration Is done.

In the drug of the present invention, curcumin or a curcumin derivative is used as an active ingredient. However, the curcumin derivative here has an orthomethoxylphenol group (o-methoxylphenol) and a β-diketone structure. The structure of demethoxycurcumin, which is a specific example of the corresponding curcumin derivative, is shown below.

This curcumin derivative (demethoxycurcumin) has a β-diketone structure like curcumin, but has an orthomethoxylphenol moiety only on one side. The structure of curcumin is shown below.

  As long as the above conditions are satisfied, various curcumin derivatives can be used as the active ingredient of the present invention. Examples of the curcumin derivative include a derivative in which a part (atom or atomic group) of the structure of curcumin is substituted with another atom or atomic group. Such curcumin derivatives can be prepared by any manufacturing process designed to yield the derivative as a final product. As other atoms or atomic groups here, hydroxyl group, halogen (fluorine, chlorine, bromine, iodine, etc.), alkyl group (methyl group, ethyl group, n-propyl group, isopropyl group, etc.), hydroxyalkyl group (hydroxymethyl) Group, hydroxyethyl group, etc.), alkoxy group (methoxy group, ethoxy group, etc.), acyl group (formyl group, acetyl group, malonyl group, benzoyl group, etc.) and the like.

    As shown in Examples described later, curcumin showed a particularly high GLP-1 secretion promoting action. Therefore, in a preferred embodiment, curcumin is employed as an active ingredient. On the other hand, as a result of comparative experiments with curcumin derivatives, it was found that the two orthomethoxylphenol sites and β-diketone structure of curcumin are important for GLP-1 secretion promoting action. A curcumin derivative having a diketone structure can be said to be a preferable active ingredient as well as curcumin.

  You may use the pharmacologically acceptable salt of curcumin or a curcumin derivative as an active ingredient of the chemical | medical agent of this invention. “Pharmaceutically acceptable salt” is to be interpreted in a broad sense and is a term that includes various salts such as acid addition salts, metal salts, ammonium salts, organic amine addition salts, amino acid addition salts, and the like. Examples of acid addition salts include inorganic acid salts such as trifluoroacetate, hydrochloride, sulfate, nitrate, phosphate, hydrobromide, acetate, maleate, fumarate, citrate, Organic acid salts such as benzene sulfonate, benzoate, malate, oxalate, methanesulfonate, and tartrate are listed. Examples of the metal salt include alkali metal salts such as sodium salt, potassium salt and lithium salt, alkaline earth metal salts such as magnesium salt and calcium salt, aluminum salt and zinc salt. Examples of ammonium salts include salts such as ammonium and tetramethylammonium. Examples of organic amine addition salts include morpholine addition salts and piperidine addition salts. Examples of amino acid addition salts include glycine addition salts, phenylalanine addition salts, lysine addition salts, aspartic acid addition salts, and glutamic acid addition salts.

  Curcumin and curcumin derivatives are components contained in turmeric (turmeric). Therefore, the active ingredient of the present invention can be extracted from turmeric. The active ingredient of the present invention may be prepared by chemical synthesis. Curcumin and curcumin derivatives extracted and purified from turmeric are commercially available and may be used.

  The agent of the present invention promotes the secretion of GLP-1 that exhibits various physiological actions. When the agent of the present invention is administered to a living body, the physiological actions of GLP-1 (promoting insulin secretion, suppressing glucagon secretion, suppressing gastric excretion and gastric acid secretion, appetite, Inhibition of feeding, promotion of pancreatic β-cell proliferation, improvement of learning / memory ability, etc. (Gastroenterology 2007, 132: 2131-57)) can be enhanced or improved. Therefore, the drug of the present invention can prevent or improve diabetes, obesity, postprandial hyperglycemia or neurodegenerative diseases such as dementia, improvement of neuropathy associated with diabetes, suppression of glucagon secretion, suppression of gastric excretion and gastric acid secretion, appetite, It is effective for suppressing feeding and promoting proliferation of pancreatic β cells. Here, “prevention” refers to preventing or delaying the onset / onset of a disease (disorder) or its symptoms, or reducing the risk of onset / onset. On the other hand, “improvement” means that a disease (disorder) or a symptom thereof is alleviated (lightened), improved, ameliorated, or cured (including partial healing).

  As described above, the agent of the present invention can be used in various applications, and a particularly important application is prevention or amelioration of diabetes and obesity. Then, the 2nd aspect of this invention provides the composition for prevention or improvement of diabetes or obesity containing the chemical | medical agent of this invention. Although the form of the composition of this invention is not specifically limited, Preferably it is a pharmaceutical, a quasi-drug, or a foodstuff. Two or more kinds of active ingredients (for example, a combination of curcumin and demethoxycurcumin) may be used in combination.

“Diabetes” is a disease characterized by a chronic increase in blood sugar (ie, hyperglycemia). It is one of the important risk factors for ischemic heart disease (angina, myocardial infarction), arteriosclerosis, cerebrovascular disorder (cerebral infarction, etc.), and is attracting attention as a representative disease of so-called “lifestyle-related diseases” . On the other hand, “obesity” generally refers to a condition in which adipose tissue is excessively accumulated in the body. In this specification, the term “obesity” is to be interpreted in a broad sense, and the concept includes obesity. “Obesity” refers to a medical condition that is or is predicted to have a health disorder (complication) due to or related to obesity, and that requires medical weight loss. As a method for determining obesity, for example, there is a method based on a BMI (body mass index) widely used internationally. BMI is a numerical value obtained by dividing body weight (kg) by the square of height (m) (BMI = weight (kg) / height (m) ² ). BMI <18.5 is underweight, 18.5 ≦ BMI <25 is normal range, 25 ≦ BMI <30 is preobese, 30 ≦ BMI <35 is obesity 2 obese class I), 35 ≦ BMI <40 is determined as obesity class II, and 40 ≦ BMI is determined as obesity class 4 (WHO). In addition, using BMI, the standard weight (ideal weight) of a Japanese adult is calculated from the following formula, standard weight (kg) = height (m) ² × 22, and the measured weight is the standard weight (calculated value). There is also a method for determining obesity in a state exceeding 120% of the above. However, since the standard weight (ideal weight) varies from individual to individual depending on sex, age, or lifestyle, etc., it is considered inappropriate to uniformly determine obesity by this method.

  The pharmaceutical composition and quasi-drug composition of the present invention can be formulated according to a conventional method. In the case of formulating, other pharmaceutically acceptable ingredients (for example, carriers, excipients, disintegrants, buffers, emulsifiers, suspension agents, soothing agents, stabilizers, preservatives, preservatives, physiological Saline solution and the like). As the excipient, lactose, starch, sorbitol, D-mannitol, sucrose and the like can be used. As the disintegrant, starch, carboxymethylcellulose, calcium carbonate and the like can be used. Phosphate, citrate, acetate, etc. can be used as the buffer. As the emulsifier, gum arabic, sodium alginate, tragacanth and the like can be used. As the suspending agent, glyceryl monostearate, aluminum monostearate, methyl cellulose, carboxymethyl cellulose, hydroxymethyl cellulose, sodium lauryl sulfate and the like can be used. As the soothing agent, benzyl alcohol, chlorobutanol, sorbitol and the like can be used. As the stabilizer, propylene glycol, ascorbic acid or the like can be used. As preservatives, phenol, benzalkonium chloride, benzyl alcohol, chlorobutanol, methylparaben, and the like can be used. As preservatives, benzalkonium chloride, paraoxybenzoic acid, chlorobutanol and the like can be used.

  The dosage form in the case of formulating is also not particularly limited, and the pharmaceutical composition or pharmaceutical of the present invention can be used as tablets, powders, fine granules, granules, capsules, syrups, injections, external preparations, suppositories, etc. An quasi-drug composition can be provided.

  The pharmaceutical composition of the present invention contains an active ingredient in an amount necessary for obtaining an expected therapeutic effect or preventive effect (that is, a therapeutically effective amount). Similarly, the quasi-drug composition of the present invention contains an active ingredient in an amount necessary for obtaining the expected improvement effect, prevention effect and the like. The amount of active ingredient contained in the pharmaceutical composition or quasi-drug composition of the present invention generally varies depending on the dosage form and form, but the amount of active ingredient is, for example, about 0.1% by weight to achieve a desired dose. Set within the range of about 95% by weight.

  The pharmaceutical composition and quasi-drug composition of the present invention are orally or parenterally (enteral, intravenous, intraarterial, subcutaneous, intramuscular or intraperitoneal injection, transdermal, nasal, Applied to the subject by transmucosal, application, etc.). The “subject” here is not particularly limited, and includes humans and non-human mammals (including pet animals, domestic animals, laboratory animals. Specifically, for example, mice, rats, guinea pigs, hamsters, monkeys, cows, pigs, goats. , Sheep, dogs, cats, chickens, quails, etc.). In a preferred embodiment, the application subject is a human.

  The dosage and usage of the pharmaceutical composition and quasi-drug composition of the present invention are set so as to obtain the expected effect. In setting an effective dose, the symptom, age, sex, weight, etc. of the subject of application are generally considered. A person skilled in the art can set an appropriate dose in consideration of these matters. As the administration schedule, for example, once to several times a day, once every two days, or once every three days can be adopted. In preparing the administration schedule, the symptom of the application target, the duration of effect of the active ingredient, and the like can be considered.

  As described above, one embodiment of the present invention is a food composition containing the drug of the present invention. Examples of the “food composition” in the present invention include general foods (cereals, vegetables, meat, various processed foods, confectionery, milk, soft drinks, alcoholic beverages, etc.), nutritional supplements (supplements, nutritional drinks, etc.), Mention may be made of food additives, foods for pets, nutritional supplements for pets. In the case of a dietary supplement or food additive, it can be provided in the form of powder, granule powder, tablet, paste, liquid or the like. By providing in the form of a food composition, it becomes easy to ingest the active ingredient of the present invention on a daily basis or continuously.

  The food composition of the present invention preferably contains an active ingredient in an amount that can be expected to have a therapeutic or prophylactic effect. The amount added can be determined in consideration of the medical condition, health status, age, sex, weight, etc. of the person to whom it is used.

<GLP-1 secretion test in cultured cells>
GLP-1-producing cell line derived from mouse colon was cultured in 10% FBS-containing DMEM (Dulbecco's modified eagle's medium, high glucose, SIGMA) in a CO ₂ incubator (37 ° C, CO ₂ concentration 5%) . It was passaged when it became 80% confluent. The cells were seeded in a 24-well plate and cultured until 80% confluent. Before adding the test sample, remove the medium from each well and replace with a buffer containing bovine serum albumin (Wako Pure Chemicals) containing 120 mM NaCl, 5 mM KCl, 2 mM CaCl ₂ , 1 mM MgCl ₂ , 22 mM NaHCO ₃ 0.5% fatty acid. And kept in a CO ₂ incubator (37 ° C., CO ₂ concentration 5%) for 1 hour. Thereafter, the buffer in each well was removed and washed with the same buffer. Thereafter, in the presence of a sample (10 mM glutamine; positive control, curcumin, curcumin derivative 2 (demethoxycurcumin), curcumin derivative 3 (bisdemethoxycurcumin), tetrahydrocurcumin) (FIG. 1), 2 in the same buffer. Incubate for hours. After 2 hours, the culture supernatant was collected in a separate tube, and the supernatant obtained by centrifugation at 800 × g and 4 ° C. for 5 minutes was stored frozen. The concentration of GLP-1 in the supernatant was measured using a commercially available Enzyme immuno assay kit (Millipore) according to the protocol attached to the kit. The statistical process used Turkey-Kramer test. P <0.05 was considered significant. Glutamine used as a positive control is known to have a GLP-1 secretion promoting action (Reimann F et al. Diabetologia (2004) 47: 1592-1601).

<Result>
The addition of curcumin (final concentration 25 μM) significantly promoted GLP-1 secretion compared to the control. The addition of curcumin derivative 2 (final concentration 25 μM) also significantly promoted GLP-1 secretion, but the level of increased secretion was lower than that of curcumin. On the other hand, addition of curcumin derivative 3 and tetrahydrocurcumin (both at a final concentration of 25 μM) did not significantly promote secretion of GLP-1 as compared to the control (FIG. 2). When the concentration of curcumin was changed (curcumin final concentrations of 10 μM, 25 μM, and 50 μM), all significantly promoted the secretion of GLP-1 compared to the control, and the amount of secretion increased as the concentration increased (FIG. 3). ).

<Discussion>
As a result of the above experiments, (1) having two o-methoxylphenol sites and having a β-diketone structure (curcumin) promotes secretion most, (2) out of two o-methoxylphenol sites , Disappearance of one methoxyl group (curcumin derivative 2 (demethoxycurcumin)) has GLP-1 secretion promoting action, but its action decreases, (3) Of the two o-methoxylphenol sites, When 2 methoxyl groups disappear (curcumin derivative 3 (bisdemethoxycurcumin)), GLP-1 secretion promoting activity disappears, (4) β-diketone structure even if it has 2 o-methoxylphenol sites (Tetrahydrocurcumin) showed that the GLP-1 secretion promoting action disappears, and the GLP-1 secretion promoting action is exhibited by the o-methoxylphenol site and β-dike It was found down structure is essential and important.

  Curcumin used as an active ingredient of the present invention is a component (naturally-derived component) contained in turmeric. The use of naturally-derived components is advantageous and important from the viewpoint of safety, and is also suitable for daily or continuous ingestion (administration). The drug of the present invention is useful for prevention and improvement of various diseases and symptoms in which promotion of GLP-1 secretion is effective. Specifically, prevention or improvement of diabetes, obesity, postprandial hyperglycemia, neurodegenerative diseases such as dementia, improvement of neuropathy associated with diabetes, suppression of glucagon secretion, suppression of gastric excretion and gastric acid secretion, appetite and eating The present invention can be used as pharmaceuticals, quasi drugs, foods, supplements, and the like.

  The present invention is not limited to the description of the embodiments and examples of the invention described above. Various modifications may be included in the present invention as long as those skilled in the art can easily conceive without departing from the description of the scope of claims. The contents of papers, published patent gazettes, patent gazettes, and the like specified in this specification are incorporated by reference in their entirety.

Claims (5)

  A GLP-1 secretion promoter comprising curcumin or a curcumin derivative (provided having an orthomethoxylphenol group and a β-diketone structure) or a salt thereof as an active ingredient. The GLP-1 secretion promoter according to claim 1, wherein the curcumin derivative is represented by the following chemical formula.

  Prevention or improvement of diabetes, obesity, postprandial hyperglycemia or neurodegenerative disease, improvement of neuropathy associated with diabetes, suppression of glucagon secretion, suppression of gastric excretion and gastric acid secretion, suppression of appetite and food intake, or promotion of pancreatic β-cell proliferation The GLP-1 secretion promoter of Claim 1 or 2 used for this.   A composition for preventing or improving diabetes or obesity, comprising the GLP-1 secretion promoter according to claim 1 or 2.   The composition according to claim 4, which is a medicine, quasi drug or food.

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