JP2013053091A - Glp-1 secretion promoter - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a substance useful for promoting GLP-1 secretion, suppressing gastric emptying, etc.SOLUTION: A GLP-1 secretion suppressor comprises spirulina or an extract thereof as an active ingredient. A gastric emptying suppressor comprises spirulina or an extract thereof as an active ingredient.

Description

  The present invention relates to a GLP-1 (glucagon-like peptide-1) secretion promoter.

  Glucagon-like peptide-1 (GLP-1) is a hormone produced from a gene and a precursor common to glucagon, which is an endocrine hormone. While glucagon is produced by α cells in the splenic islets, GLP-1 is produced by intestinal endocrine cells (L cells), and is produced from the precursor through processing different from that of glucagon. GLP-1 is a hormone with an incretin (Intestine Secretion Insulin) action that plays an important role in glucose metabolism. It is secreted into the blood by ingestion of nutrients and acts on the pancreatic β-cells to promote insulin secretion. , Lower blood sugar levels. GLP-1 also acts on the pancreas to suppress glucagon secretion, thereby reducing glucose release from the liver and lowering blood glucose levels. Other actions of GLP-1 are known to promote pancreatic β-cell proliferation, suppress gastric excretion and gastric acid secretion, and suppress appetite and food intake (Non-Patent Documents 1 to 3). Therefore, enhancing the effect of GLP-1 is useful for improving lifestyle-related diseases such as obesity and diabetes. In recent years, development of diabetes treatment methods using GLP-1 replacement therapy and GLP-1 receptor (GLP-1R) agonists has also been promoted.

  GLP-1 is also known to have an effect on the central nervous system. GLP-1's appetite and feeding-suppressing effects are thought to be via GLP-1R present in the central nervous system (Non-patent Document 1). Moreover, while learning ability decreases in a GLP-1R-deficient mouse, memory learning ability improves in a mouse in which GLP-1R is overexpressed (Non-patent Document 4). In experiments in a culture system, GLP-1 has been observed to promote neurite outgrowth and to protect against apoptosis due to nerve injury (Non-patent Document 5). Therefore, GLP-1 is expected to be a useful substance for suppressing neurodegeneration and improving neurodegenerative diseases such as dementia and neuropathy associated with diabetes.

  GLP-1 is also considered useful for controlling blood glucose levels in diabetic patients. When GLP-1 suppresses pancreatic glucagon secretion, glucose release from the liver decreases and blood glucose decreases. It has been reported that this action does not antagonize glucagon secretion due to hypoglycemia and does not depend on insulin concentration. It has also been reported that administration of GLP-1 decreased the amount of insulin required to make blood glucose constant in both type 1 diabetic patients and type 2 diabetic patients. Furthermore, GLP-1 can exert an effect on blood glucose level control in the long term by acting on the central nervous system and suppressing appetite. Therefore, GLP-1 may improve the symptoms of patients with diabetes and insulin resistance through these actions. Exenatide and Liraglutide, which are actually GLP-1 analog preparations, have been used as antidiabetic drugs and have been reported to have an effect of improving insulin resistance (Non-patent Document 6).

  However, GLP-1 is usually very susceptible to degradation and has a very short half-life in vivo. Therefore, even if GLP-1 is administered from outside the body, it is very difficult to keep the blood concentration constant. Therefore, in order to increase the in vivo GLP-1 concentration over a long period of time, it is desirable to promote the secretion of endogenous GLP-1 rather than external administration.

  As a substance that promotes secretion of GLP-1 from research in a culture system, palmitic acid, oleic acid, meat hydrolyzate (MH), gastrin releasing peptide (GRP), carbachol, forskolin, ionomycin, phorbol myristate acetate (PMA), essential amino acids (EAA), leucine, isoleucine, skim milk, casein, leptin, and muscarinic receptors M1 and M2 have been reported so far (Non-Patent Documents 7 to 11).

  Spirulina is an algae belonging to the genus Cyanobacteria genus Ursulospira, and is known to have a glycohemoglobin lowering action and a serum lipid level improving action (Non-patent Document 12). However, there is no report on the action of Spirulina on GLP-1.

J Clin Endocrinol Metab, 2002, 87 (3): 1239-1246 Am J Physiol Endocrinol Metab, 1997, 273: E981-E988 Eur J Pharmacol, 2002, 440 (2-3): 269-279 Nat Med, 2003, 9: 1173-1179 J Pharmacol Exp Ther, 2002, 302 (3): 881-888 History of Medicine, 2009, 231 (7): 755-758 Endocrinology, 2001, 142 (10): 4522-4528 Journal of Endocrinology, 2006, 191,159-170 Nutrition, 2009, 25 (3): 340-349 Endocrinology, 2003, 144 (7): 3244-3250 Diabetes, 2006, 52 (2): 252-259 Journal of Medicinal Food, 2001, 4 (4): 193-199

  The present invention relates to the provision of a substance that promotes secretion of GLP-1, and the use of the substance in connection with promotion of GLP-1 secretion, suppression of gastric excretion, and the like.

  When the present inventors examined the material which can accelerate | stimulate the secretion of GLP-1, it discovered that Spirulina had the effect | action which accelerates | stimulates the secretion of GLP-1.

That is, the present invention provides the following.
(1) A GLP-1 secretion inhibitor comprising spirulina or an extract thereof as an active ingredient.
(2) A gastric excretion inhibitor comprising spirulina or an extract thereof as an active ingredient.

  The GLP-1 secretion promoter of the present invention promotes excellent GLP-1 secretion and is useful for suppressing gastric excretion.

Promotion of GLP-1 secretion in NCI-H716 cells by Spirulina extract. *: P <0.05.

  As used herein, “GLP-1 secretion promotion” means promoting GLP-1 secretion in vivo caused by ingesting a meal, particularly a meal rich in carbohydrates or lipids. Alternatively, “promoting GLP-1 secretion” refers to enhancing the increase in blood GLP-1 concentration accompanying GLP-1 secretion in vivo, mainly occurring after meals, or maintaining the increased GLP-1 concentration, Suppressing a decrease in the increased GLP-1 concentration.

  In the present specification, “non-therapeutic” is a concept that does not include a medical act, that is, a treatment act on the human body by therapy.

  As used herein, “improvement” refers to improvement of a disease, symptom or condition, prevention or delay of worsening of the disease, symptom or condition, or reversal, prevention or delay of progression of a disease or symptom.

  As used herein, “prevention” refers to preventing or delaying the onset of a disease or symptom in an individual, or reducing the risk of developing an individual's disease or symptom.

In the present specification, “spirilina” refers to Arthrospira platensis or Arthrospira maxima of the cyanobacterium Lepidoptera. The “Spirulina extract” may be an extract from Spirulina whole algae. Spirulina may be subjected to the extraction process as it is, or may be subjected to the extraction process after being pulverized, cut or dried.

  As said extract, what is marketed, for example, a commercially available spirulina pigment | dye, may be utilized, or various solvent extracts obtained by a conventional method, its dilution liquid, its concentrate, its dry powder, paste Or the activated carbon treatment may be used. As an example, the extract in the present invention is obtained by extracting Spirulina at room temperature (for example, 4 to 50 ° C.) or under heating (room temperature to solvent boiling point), or using an extraction device such as a Soxhlet extractor. Can be obtained.

  As the solvent for extraction, either a polar solvent or a nonpolar solvent can be used. Specific examples of the solvent include water; alcohols such as methanol, ethanol, propanol, and butanol; polyhydric alcohols such as propylene glycol and butylene glycol; ketones such as acetone and methyl ethyl ketone; methyl acetate, ethyl acetate, and the like. Esters; linear and cyclic ethers such as tetrahydrofuran and diethyl ether; polyethers such as polyethylene glycol; hydrocarbons such as squalane, hexane, cyclohexane and petroleum ether; aromatic hydrocarbons such as toluene; dichloromethane and chloroform And halogenated hydrocarbons such as dichloroethane; and supercritical carbon dioxide; pyridines; organic solvents such as fats and waxes and other oils; and mixtures thereof. Preferable examples include water, alcohols and aqueous solutions thereof, and ethanol is preferable as the alcohols. More preferred solvents are water and an aqueous ethanol solution.

In the case of using an aqueous alcohol solution as a solvent for extraction, the mixing ratio (volume ratio) of alcohols and water is preferably 0.001 to 100: 99.999-0, and 5 to 95:95 to 5 is more preferable, 20 to 80:80 to 20 is more preferable, 30 to 70:70 to 30 is still more preferable, and 40 to 60:60 to 40 is still more preferable. In the case of an ethanol aqueous solution, the ethanol concentration is preferably 40 to 60% by volume.
As a usage-amount of a solvent, 1-100 mL is preferable with respect to 1 g of Spirulina (dry mass conversion), As extraction time, 1 minute-100 days are preferable, and 1 minute-10 days are more preferable. The extraction temperature at this time is 0 ° C. to the boiling point of the solvent, more preferably 5 to 100 ° C., further preferably 10 to 50 ° C., and still more preferably 10 to 35 ° C.

Specifically, the extraction means for obtaining the Spirulina extract uses means such as solid-liquid extraction, liquid-liquid extraction, immersion, decoction, leaching, steam distillation, reflux extraction, ultrasonic extraction, microwave extraction, and stirring. be able to. For example, as a suitable example of immersion, immersion at 10 to 50 ° C. for 1 hour to 14 days can be mentioned. Moreover, when shortening extraction time, solid-liquid extraction with stirring is desirable. An example of suitable conditions for this solid-liquid extraction is stirring at 100 to 400 rpm under 10 to 100 ° C. (preferably 10 to 35 ° C.).
In order to prevent the oxidation of the extract, a means for extracting under a so-called non-oxidizing atmosphere while removing dissolved oxygen by bubbling degassing or inert gas such as nitrogen gas may be used in combination.

  As shown in Examples below, Spirulina extract has an effect of significantly promoting GLP-1 secretion from gastrointestinal cells. In addition, as an action of GLP-1, glucagon secretion suppression, insulin secretion promotion, suppression of blood glucose increase, gastric excretion suppression, and appetite suppression are well known (J Clin Endocrinol Metab, 2002, 87 ( 3): 1239-1246, Am J Physiol Endocrinol Metab, 1997, 273: E981-E988, Eur J Pharmacol, 2002, 440 (2-3): 269-279). Therefore, Spirulina or an extract thereof is useful as a GLP-1 secretion promoter and is useful for controlling various states related to GLP-1 secretion, for example, suppressing gastric excretion.

  That is, Spirulina or an extract thereof can be used for promoting GLP-1 secretion or for suppressing gastric excretion. The use can be in humans or non-human animals, or specimens derived therefrom, and can be therapeutic or non-therapeutic.

Accordingly, the present invention provides a GLP-1 secretion promoter containing spirulina or an extract thereof as an active ingredient.
The present invention also provides a gastric excretion inhibitor containing spirulina or an extract thereof as an active ingredient.
The agent of the present invention may consist essentially of Spirulina or an extract thereof.

  Spirulina or an extract thereof may be blended as a raw material in a composition for promoting GLP-1 secretion or suppressing gastric excretion, a medicine, a quasi-drug, a food or drink, or a raw material thereof, or a feed or a raw material thereof. Or they can be used for their manufacture.

  The composition, medicine, quasi-drug, food / beverage product, feed, or food / beverage product or feed material may be produced or used for human or non-human animals. Spirulina or its extract is blended as a raw material of the composition, medicine, quasi-drug, food and drink, feed, or food or drink or feed, and is an active ingredient for promoting GLP-1 secretion or suppressing gastric excretion It can be.

  The medicine or quasi-drug contains spirulina or an extract thereof as an active ingredient. The pharmaceutical or quasi drug can be administered in any dosage form. The dosage form may be oral or parenteral. For example, oral dosage forms include solid dosage forms such as tablets, coated tablets, granules, powders, capsules, and liquid dosage forms such as elixirol, syrups and suspensions, and parenteral dosage forms. Examples thereof include injection, infusion, transdermal, transmucosal, nasal, enteral, inhalation, suppository, bolus, and patch.

  The said medicine and quasi-drug may contain Spirulina or an extract thereof alone or in combination with a pharmaceutically acceptable carrier. Examples of such carriers include excipients, coating agents, binders, extenders, disintegrants, surfactants, lubricants, diluents, dispersants, buffers, osmotic pressure adjusting agents, pH adjusting agents, Examples include emulsifiers, preservatives, stabilizers, antioxidants, colorants, ultraviolet absorbers, humectants, thickeners, activity enhancers, anti-inflammatory agents, bactericides, taste-masking agents, and flavoring agents. Moreover, the said pharmaceutical and quasi-drug may contain another active ingredient and a pharmacological component, unless the GLP-1 secretion promotion effect of spirulina or its extract is lost.

  The pharmaceutical or quasi-drug can be produced from Spirulina or an extract thereof, or in combination with the above-mentioned carrier and / or other active ingredients and pharmacological ingredients as required by a conventional method. The content of spirulina in the medicine or quasi-drug (in terms of dry matter of the extract) is usually 0.0001 to 20% by mass, preferably 0.001 to 10% by mass, and 0.01 to More preferably, it is 5 mass%.

  The above-mentioned food and drink and feed have functions such as promotion of GLP-1 secretion and suppression of gastric excretion, and food, functional food, food for the sick, food for specified health use, pet food displaying such functions as necessary Etc.

  The kind of said food / beverage products is not specifically limited. Examples of the beverage include all beverages such as fruit juice beverages, carbonated beverages, tea beverages, coffee beverages, milk beverages, alcoholic beverages, and soft drinks. The form of the food may be any form such as solid, semi-solid, liquid, etc., and may be tablet form, pill form, tablet, capsule form, liquid form, syrup form, powder form, granule form, etc. Also good. For example, as food, breads, noodles, pasta, jelly-like foods, various snacks, cakes, confectionery, ice creams, soups, dairy products, frozen foods, instant foods, other processed foods, seasonings, And supplements. The kind of the feed is not particularly limited and may be a feed for any animal, and the form thereof may be any form as in the case of the food.

  The above-mentioned food and drink or feed, or their raw materials may contain spirulina or an extract thereof alone, or other foods, solvents, softeners, oils, emulsifiers, preservatives, aromatics, stabilizers. Further, additives such as a colorant, an antioxidant, a moisturizing agent, and a thickener may be contained in combination. The content of spirulina in the food or drink or feed (in terms of dry matter of the extract) is usually 0.0001 to 10% by mass, preferably 0.0001 to 5% by mass, and 0.001 to 1 It is more preferable to set it as the mass%.

  Alternatively, Spirulina or an extract thereof is administered in an effective amount to a subject in need thereof for promoting GLP-1 secretion or suppressing gastric excretion. Alternatively, Spirulina or an extract thereof is taken in an effective amount by a subject in need thereof for promoting GLP-1 secretion or suppressing gastric excretion.

  The subject to be administered or ingested may be an animal that requires GLP-1 secretion promotion or gastric excretion suppression. The animal is preferably a human or non-human mammal, more preferably a human.

  Preferred dosages and intakes may vary according to the subject's species, weight, sex, age, condition or other factors. The dose, route, interval, and amount and interval of intake can be determined appropriately by those skilled in the art. For example, in the case of oral administration or ingestion to humans, the administration or intake is preferably 20 μg to 2000 mg / day, and 200 μg to 1000 mg / day per adult as the amount of spirulina (in terms of the dried product of the extract). Is more preferable, 2 mg to 500 mg / day is more preferable, and 20 mg to 200 mg / day is still more preferable. Oral administration or ingestion is preferably performed at the time of each meal or before each meal, more preferably 4 hours to 5 minutes before each meal.

  Alternatively, the subject to be administered or ingested can be an animal-derived tissue, organ, cell, or fraction thereof. The tissue, organ, cell, or fraction thereof is preferably a naturally derived or biologically or biotechnologically modified tissue, organ, cell, or their ability to secrete GLP-1. It is a fraction.

  EXAMPLES Hereinafter, an Example is shown and this invention is demonstrated more concretely.

Example 1 GLP-1 Secretion Promoting Action in Spirulina Cells Commercial Spirulina extract (Lina Blue AE; Spirulina dye sold by Dainippon Ink, Inc.) is adjusted to a concentration of 1% (w / v) with 20% EtOH. To obtain a test sample. As a control, 20% EtOH was used.

NCI-H716 cells (human colon-derived cells; purchased from ATCC, developed by Dr. Herbert K. Oie et al. And supplied by agencies of the United States Public Health Service) were cultured at 37 ° C. in the presence of 5% CO ₂ . . As the culture solution, RPMI1640 (containing 10% fetal calf serum, high glucose; Invitrogen) was used. Cells were seeded in a 24-well plate coated with Matrigel (100 μL / well; BD) to 1 × 10 ⁶ cells / well (N = 4), and DMEM (containing 10% fetal bovine serum, high glucose; Invitrogen) Incubated in medium. Three days later, the medium was replaced with KRB (Krebs-Ringer bicarbonate, Sigma) /0.2% BSA containing the test sample at a concentration of 0.01% (v / v) and cultured for 2 hours, and then the medium was recovered. did. The medium was collected in a microcentrifuge tube to which diprotin-A (DPP4 inhibitor, Sigma) and PMSF (Phenylmethylsulfonyl fluoride, serine protease inhibitor, Sigma) were added. Stored at -80 ° C. GLP-1 in the medium was quantified by ELISA using GLUCAGON-LIKE PEPTIDE-1 (ACTIVE) ELISA KIT (LINCO Research) and determined as a relative value to the average value of the control.

  The results are shown in FIG. Spirulina significantly promoted GLP-1 secretion at a concentration of 0.01%.

Claims (2)

  A GLP-1 secretion inhibitor comprising spirulina or an extract thereof as an active ingredient.   A gastric excretion inhibitor comprising spirulina or an extract thereof as an active ingredient.

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