WO2010119674A1 - レプチン抵抗性を改善および/または予防するための薬学的組成物、並びにその使用 - Google Patents
レプチン抵抗性を改善および/または予防するための薬学的組成物、並びにその使用 Download PDFInfo
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- WO2010119674A1 WO2010119674A1 PCT/JP2010/002691 JP2010002691W WO2010119674A1 WO 2010119674 A1 WO2010119674 A1 WO 2010119674A1 JP 2010002691 W JP2010002691 W JP 2010002691W WO 2010119674 A1 WO2010119674 A1 WO 2010119674A1
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
- A61K31/19—Carboxylic acids, e.g. valproic acid
- A61K31/192—Carboxylic acids, e.g. valproic acid having aromatic groups, e.g. sulindac, 2-aryl-propionic acids, ethacrynic acid
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/04—Anorexiants; Antiobesity agents
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/06—Antihyperlipidemics
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
- A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61P9/00—Drugs for disorders of the cardiovascular system
- A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61P9/00—Drugs for disorders of the cardiovascular system
- A61P9/12—Antihypertensives
Definitions
- the present invention relates to a pharmaceutical composition for improving and / or preventing leptin resistance and use thereof.
- Leptin is a hormone that is secreted mainly by adipose tissue. Leptin secreted into the circulating blood reduces body weight by causing an antifeedant action and increased energy consumption via leptin receptors present in the hypothalamus and brainstem.
- Non-Patent Documents 2, 6 and 7 show that protein tyrosine phosphatase 1B is involved in leptin resistance.
- Non-patent documents 3 and 4 show that leptin resistance is induced by increased expression of suppressor ⁇ cytokine signaling 3 (SOCS3) in the hypothalamus. Furthermore, in Non-Patent Document 5, rather than the reactivity of leptin itself, there is some cause in the system that transports leptin in circulating blood to the central system through the blood-brain barrier and induces leptin resistance. It has been suggested.
- SOCS3 suppressor ⁇ cytokine signaling 3
- leptin resistance is considered to be a cause of onset of diabetes, hypertension, hyperlipidemia, arteriosclerosis, etc., and improving and / or preventing leptin resistance It is also important for ameliorating and / or preventing diseases caused by sex.
- Patent Document 1 discloses a pharmaceutical composition for improving endoplasmic reticulum stress in the central nervous system. Such pharmaceutical compositions are characterized by containing non-steroidal anti-inflammatory compounds. It has been suggested that flurbiprofen can be used as the non-steroidal anti-inflammatory compound.
- Japanese Patent Publication Japanese Patent Laid-Open No. 2008-208091 (published on September 11, 2008)”
- Patent Document 1 is intended to improve endoplasmic reticulum stress in the central nervous system, and has been developed in consideration of improving and / or preventing leptin resistance. There is no.
- leptin resistance is induced by various causes, and the fundamental mechanism of leptin resistance is still unclear. Therefore, a definitive therapeutic drug for improving and / or preventing leptin resistance has not been obtained.
- the present invention has been made in view of the above problems, and an object thereof is to provide a pharmaceutical composition for improving and / or preventing leptin resistance, and use of the pharmaceutical composition. It is in.
- flurbiprofen which has been conventionally applied to diseases such as rheumatoid arthritis and low back pain as a nonsteroidal anti-inflammatory drug, is leptin resistant. Has been found to have an effect of improving and / or preventing the present invention, and the present invention has been completed.
- flurbiprofen has an effect of improving and / or preventing leptin resistance. Therefore, the present inventors have discovered for the first time that flurbiprofen has an effect of improving and / or preventing leptin resistance, and are disclosed herein.
- the pharmaceutical composition for improving and / or preventing leptin resistance according to the present invention is characterized by containing flurbiprofen.
- the present invention encompasses the use of flurbiprofen for the manufacture of a pharmaceutical composition for improving and / or preventing leptin resistance.
- Patent Document 1 shows that the expression of GRP78 and CHOP induced by endoplasmic reticulum stress is suppressed by flurbiprofen. This only suggests that flurbiprofen can be used to improve endoplasmic reticulum stress. As described above, leptin resistance is also induced by causes other than endoplasmic reticulum stress. Therefore, it is suggested in Patent Document 1 that flurbiprofen has an effect of improving endoplasmic reticulum stress that contributes to the acquisition of leptin resistance, and thus leptin resistance to flurbiprofen is improved. It cannot be expected that there will be an effect. Therefore, the present invention cannot be easily conceived based on the description in Patent Document 1 and the like.
- the pharmaceutical composition according to the present invention exhibits a very excellent effect that leptin resistance can be improved and / or prevented.
- flurbiprofen contained in the pharmaceutical composition according to the present invention has already been marketed as a nonsteroidal anti-inflammatory drug. Therefore, side effects (stomach discomfort, loss of appetite, nausea, etc.) due to administration can be predicted, and it has also been confirmed that dependency on flurbiprofen and development of tolerance are not observed. Therefore, it is excellent in that it can be a safe drug that can be administered for a long period of time for improving and / or preventing leptin resistance.
- FIG. 6A shows the leptin concentration in plasma before fasting
- FIG. 6B shows the leptin concentration in plasma after fasting.
- FIG. 7A shows the correlation between leptin concentration in plasma before fasting and body weight
- FIG. 7B shows the correlation between leptin concentration in plasma after fasting and body weight.
- FIG. 10 is a diagram showing an experimental procedure of Example 7. It is a graph which shows the feed rate change rate of the mouse
- the pharmaceutical composition according to the present invention is a pharmaceutical composition for improving and / or preventing leptin resistance, and is characterized by containing flurbiprofen.
- flurbiprofen includes compound (I) represented by the following general formula and enantiomers thereof, and pharmaceutically acceptable salts thereof.
- flurbiprofen includes a derivative having the skeleton of the compound (I).
- the “derivative” may have the same activity as that of the compound (I) in the form of the derivative and is inactive in the form of the derivative. It may be in the form of a so-called “prodrug” that is converted to the above compound (I) and reacquires the activity by a reaction such as above. Examples of such “derivatives” include derivatives in which the carboxylic acid in the compound (I) is esterified; derivatives in which the carboxylic acid in the compound (I) is acetylated.
- Examples of the derivative in which the carboxylic acid in the compound (I) is esterified include a derivative in which the carboxylic acid in the compound (I) is methyl esterified, ethyl esterified, or phenyl esterified. .
- the pharmaceutical composition according to the present invention can be produced by a known method in the pharmaceutical field.
- the content of flurbiprofen in the pharmaceutical composition according to the present invention the formulation form, administration method, purpose of use of the pharmaceutical composition and the age, weight, symptom, etc. of the patient to whom the pharmaceutical is administered are considered. And if it is a quantity which can administer flurbiprofen within the dosage range mentioned later using the said pharmaceutical composition, it will not specifically limit.
- the lower limit of the dose of flurbiprofen is preferably 0.013 to 401 mg / kg, more preferably 0.13 to 40 mg / kg, still more preferably 0 per day for an adult (body weight 60 kg). .44-12 mg / kg.
- the preferable dose range varies depending on various conditions, an amount smaller than the above-mentioned dose may be sufficient, or an amount exceeding the above-mentioned range may be necessary.
- “leptin resistance” means a state in which leptin is present in blood but leptin signal in the hypothalamus is suppressed.
- Leptin signal acts on a leptin receptor belonging to the family of cytokine receptors to activate Jak2.
- the transcription factor STAT3 is phosphorylated.
- Phosphorylated STAT3 translocates into the nucleus and activates transcription such as pro-opiomelanocortin (POMC) (H. Munzberg, M. G. Myers, Jr., Molecular and anatomical determinants of central leptin resistance. Nature Neuroscience, 2005, 8: 566-570). Therefore, it is possible to confirm whether or not the leptin signal is suppressed using the STAT3 phosphorylation state in the tissue expressing the leptin receptor after leptin administration as an index.
- POMC pro-opiomelanocortin
- preventing leptin resistance means, for example, leptin in the hypothalamus even under conditions that induce leptin resistance (for example, a high-fat diet described below is given for a long period of time). It is intended to maintain the phosphorylated state of STAT3.
- “improvement of leptin resistance” is intended to mean that STAT3 is phosphorylated again from a state in which leptin resistance is induced and phosphorylation of STAT3 by leptin is suppressed in the hypothalamus, for example. is doing.
- prevention of leptin resistance or “improvement of leptin resistance” is not limited to the phosphorylated state of STAT3 but also includes body weight, blood glucose level and blood leptin concentration as indices. Can be evaluated. Specifically, for example, even if conditions under which leptin resistance is induced (for example, giving a high-fat diet described below for a long period of time), a state where the blood leptin concentration does not significantly increase is maintained. It can be evaluated that leptin resistance was prevented.
- the leptin resistance was induced (for example, the state in which the leptin resistance was induced by giving a high fat diet described later and the blood leptin concentration was increased), the blood leptin concentration was significantly decreased. If it becomes a state, it can be evaluated that leptin resistance has improved.
- the pharmaceutical composition according to the present invention may further contain other components (such as a pharmaceutically acceptable carrier) other than flurbiprofen that do not inhibit the effect of flurbiprofen.
- other components such as a pharmaceutically acceptable carrier
- the term “pharmaceutically acceptable carrier” (hereinafter, also simply referred to as “carrier”) is used for the purpose of assisting prescription when producing a pharmaceutical or an agricultural chemical such as animal medicine. A substance that does not have a harmful effect on active ingredients. Furthermore, it is intended that there is no toxicity in an individual who has received the pharmaceutical composition according to the present invention and that the carrier itself does not induce the production of harmful antibodies.
- various organic or inorganic carrier substances that can be used as a preparation material can be used, and can be appropriately selected according to the administration form and dosage form of a pharmaceutical composition described later.
- excipient examples include lactose, sucrose, D-mannitol, xylitol, sorbitol, erythritol, starch, crystalline cellulose and the like, but are particularly limited as long as they are usually used in the pharmaceutical field. It is not something.
- lubricant examples include magnesium stearate, calcium stearate, wax, talc, colloidal silica and the like, but are not particularly limited as long as they are usually used in the pharmaceutical field.
- binder examples include pregelatinized starch, methylcellulose, crystalline cellulose, sucrose, D-mannitol, trehalose, dextrin, hydroxypropylcellulose, hydroxypropylmethylcellulose, polyvinylpyrrolidone, etc. If it is normally used, it will not specifically limit.
- disintegrant examples include starch, carboxymethylcellulose, low-substituted hydroxypropylcellulose, carboxymethylcellulose calcium, croscarmellose sodium, carboxymethylstarch sodium, etc., which are usually used in the pharmaceutical field. If it is, it will not specifically limit.
- solvent examples include water for injection, alcohol, propylene glycol, macrogol, sesame oil, corn oil, tricaprylin and the like, but are particularly limited as long as they are usually used in the pharmaceutical field. is not.
- dissolution aid examples include polyethylene glycol, propylene glycol, D-mannitol, trehalose, benzyl benzoate, ethanol, trisaminomethane, cholesterol, triethanolamine, sodium carbonate, sodium citrate and the like. However, it is not particularly limited as long as it is usually used in the pharmaceutical field.
- surfactants such as stearyltriethanolamine, sodium lauryl sulfate, laurylaminopropionic acid, lecithin, benzalkonium chloride, benzethonium chloride, glyceryl monostearate, polyvinyl alcohol
- hydrophilic polymers such as polyvinylpyrrolidone, sodium carboxymethylcellulose, methylcellulose, hydroxymethylcellulose, hydroxyethylcellulose, and hydroxypropylcellulose, but are not particularly limited as long as they are usually used in the pharmaceutical field.
- Examples of the “isotonic agent” include sodium chloride, glycerin, D-mannitol and the like, but are not particularly limited as long as they are usually used in the pharmaceutical field.
- buffering agent examples include buffers such as phosphates, acetates, carbonates, citrates and the like, but are particularly limited as long as they are usually used in the pharmaceutical field. is not.
- analgesic agent examples include benzyl alcohol and the like, but are not particularly limited as long as they are usually used in the pharmaceutical field.
- preservative examples include p-hydroxybenzoates, chlorobutanol, benzyl alcohol, phenethyl alcohol, dehydroacetic acid, sorbic acid, and the like, but are particularly limited as long as they are usually used in the pharmaceutical field. Is not to be done.
- antioxidant examples include sulfite and ascorbic acid, but are not particularly limited as long as they are usually used in the pharmaceutical field.
- the stabilizer and flavoring agent are not particularly limited as long as they are usually used in the pharmaceutical field.
- the pharmaceutical composition according to the present invention may be administered orally or parenterally, intravenously, rectally, intraperitoneally, intramuscularly or subcutaneously. Alternatively, it can be administered by an appropriate administration route depending on the preparation form. Especially, it is preferable that the composition concerning this invention is orally administered from the reason that administration is easy.
- parenteral refers to modes of administration including intraventricular, intravenous, intramuscular, intraperitoneal, intrasternal, subcutaneous, and intraarticular injection and infusion.
- oral preparation examples of the dosage form of such a pharmaceutical composition
- examples of the dosage form of such a pharmaceutical composition include powders, granules, tablets, liposomes, and capsules. (Including soft capsules and microcapsules), solid preparations such as powders, and liquid preparations such as syrups.
- liquid preparation is produced by a method usually used in the pharmaceutical field using, for example, water; an organic solvent such as glycerol, glycol, polyethylene glycol, etc .; a mixture of these organic solvents and water as the carrier. be able to.
- the liquid preparation may further contain a solubilizing agent, a buffer, a tonicity agent, a stabilizer and the like.
- solid preparation can be produced by a method usually used in the pharmaceutical field using, for example, an excipient, a lubricant, a binder, a disintegrant, a stabilizer, a flavoring agent and the like as the carrier. it can.
- lubricants When preparing such oral preparations, lubricants, fluidity promoters, colorants, fragrances and the like may be further blended depending on the purpose.
- parenteral agent examples of the dosage form of such a pharmaceutical composition
- examples of the dosage form of such a pharmaceutical composition include injections, suppositories, and pellets. , Drops and the like.
- a parenteral agent is prepared by converting a pharmaceutical composition according to the present invention into a diluent (eg, distilled water for injection, physiological saline, aqueous glucose solution, vegetable oil for injection, sesame oil, peanut oil, Soybean oil, corn oil, propylene glycol, polyethylene glycol, etc.) and can be prepared by further adding bactericides, stabilizers, tonicity agents, soothing agents, etc., depending on the purpose. .
- a diluent eg, distilled water for injection, physiological saline, aqueous glucose solution, vegetable oil for injection, sesame oil, peanut oil, Soybean oil, corn oil, propylene glycol, polyethylene glycol, etc.
- a sustained-release preparation can be obtained by a technique usually used in the pharmaceutical field.
- the pharmaceutical composition according to the present invention may be administered alone or in combination with other drugs.
- a method of administration in combination for example, it may be administered simultaneously as a mixture with other drugs, may be administered simultaneously or concurrently as a drug different from other drugs, or may be performed over time.
- the present invention is not limited to this.
- the number of times the pharmaceutical composition according to the present invention is administered per day is not particularly limited. As long as the dosage of flurbiprofen is within the required dosage range per day, it may be administered once per day, or may be administered in multiple doses.
- the pharmaceutical composition according to the present invention is applicable to mammals other than humans (for example, mice, rats, rabbits, dogs, cats, cows, horses, pigs, monkeys, etc.). Those skilled in the art can easily understand.
- mice 4 weeks old male mice (C57BL / 6BLCr Slc, Shimizu Experimental Materials Co., Ltd.) were used as experimental animals. All animal experiments in this example were conducted according to NIH guidelines for breeding and using laboratory animals (NIH Guide for Care and Use of Laboratory Animals).
- Example 1 ⁇ Effect of flurbiprofen on leptin resistance 1> (Method for inducing and confirming leptin resistance) First, the leptin resistance induction method and the leptin resistance confirmation method used in this example will be described below.
- FIG. 1 is a graph showing changes in body weight in mice fed with a high fat diet or a normal diet.
- Statistical analysis was performed using t-test.
- “**” described in FIG. 1 has a significant difference between the weight of a group fed with a high fat diet and the weight of a group fed with a normal diet at a risk rate of less than 1%.
- “***” represents that there is a significant difference between the weight of the group fed with the high fat diet and the weight of the group fed with the normal diet at a risk rate of less than 0.1%.
- mice fed a high fat diet 60 kcal% fat, D12492, manufactured by Research DIET
- mice fed a normal diet 10 kcal% fat, D12450B, manufactured by Research CHDIET
- mice were prepared by dissolving leptin derived from mice (450-31, manufactured by PeproTech or 498-OB, rmeptin, manufactured by R & D) in physiological saline.
- leptin derived from mice 450-31, manufactured by PeproTech or 498-OB, rmeptin, manufactured by R & D
- physiological saline was administered parenterally via the tail vein so that the dose was 5 ml / kg (1 mg / kg).
- an equal volume of physiological saline was administered into the tail vein.
- FIG. 2 shows the results of examining STAT3 phosphorylation in the hypothalamus of mice 30 minutes after administration of leptin by Western blotting using an antibody (9131S, manufactured by Cell Signaling) that recognizes phosphorylated STAT3.
- STAT3 phosphorylation in the hypothalamus was suppressed in mice at 16 weeks after feeding a high fat diet even after leptin administration. This is a result showing that the leptin signal in the hypothalamus is suppressed even after administration of leptin, that is, leptin resistance is induced.
- leptin resistance can be induced by giving a high fat diet for a long period of time. Furthermore, it was confirmed that the increase in body weight of the mouse was correlated with the suppression of STAT3 phosphorylation in the hypothalamus. This indicates that the presence or absence of leptin resistance can also be evaluated by increasing the body weight of the mouse. Therefore, in this example, leptin resistance was induced by feeding a high-fat diet over a long period of time, and that leptin resistance was improved and / or prevented by administration of flurbiprofen. As an indicator.
- mice were divided into 4 groups of 7 to 8 mice, and the following meals were given to each group of mice.
- Group 1 Normal diet (10 kcal% fat, D12450B, manufactured by Research DIET) only
- Group 2 High fat diet (60 kcal% fat, D12492, manufactured by Research DIET) only
- Group 3 Normal diet + flurbiprofen (F8514, SIGMA)
- Group 4 High fat diet + flurbiprofen (F8514, manufactured by SIGMA)
- flurbiprofen was diluted with drinking water so as to be a dose of about 10 mg / kg per day, and orally administered as drinking water. Thereafter, the weight of the mice was measured every 7 days.
- FIG. 3 is a graph showing changes in body weight of mice in each group for 8 weeks after administration of flurbiprofen.
- Statistical analysis was performed using t-test.
- “*” shown in the graph indicates that there is a significant difference between the weight of the group 2 and the weight of the group 4 when the risk rate is less than 5%.
- Example 2 ⁇ Effect of flurbiprofen on leptin resistance 2> (Method for inducing and confirming leptin resistance)
- a normal diet (10 kcal% fat, D12450B, Research DIET) or a high fat diet (60 kcal% fat, D12492, Research DIET).
- Flurbiprofen was diluted with drinking water so that the dose was about 10 mg / kg per day, and was orally administered as drinking water.
- mice were divided into 4 groups of 7 to 8 animals, and the following meals were given to the mice of each group.
- Group 1 Normal diet (10 kcal% fat) only
- Group 2 High fat diet (60 kcal% fat) only
- Group 3 Normal diet + flurbiprofen
- Group 4 High fat diet + flurbiprofen High fat diet or normal diet 8 weeks old mice were prepared by dissolving leptin derived from mouse (450-31, manufactured by PeproTech or 498-OB, rmeptin, R & D) in physiological saline, and administered at a dose of 1 mg. / Kg was administered parenterally via the tail vein. As a control for leptin administration, an equal volume of physiological saline was administered into the tail vein.
- FIG. 4 and FIG. 5 show the results of examining STAT3 phosphorylation in the hypothalamus of mice 30 minutes after administration of leptin by Western blotting using an antibody (9131S, manufactured by Cell Signaling) that recognizes phosphorylated STAT3.
- FIG. 4 is a graph showing the results of western blotting of STAT3 phosphorylation in the hypothalamus of mice of each group 30 minutes after administration of leptin.
- FIG. 5 is a graph in which the results of the Western blot shown in FIG. 4 are digitized.
- the vertical axis in FIG. 5 represents the intensity of STAT3 phosphorylation in the hypothalamus of each group of mice.
- the intensity of STAT3 phosphorylation in the hypothalamus of the group 4 mice fed with the high fat diet + flurbiprofen is similar to that of the group 2 mice fed with the high fat diet. Significantly stronger compared to the intensity of STAT3 phosphorylation at the bottom. From these results, by simultaneously administering a high-fat diet and flurbiprofen, suppression of STAT3 phosphorylation, that is, induction of leptin resistance caused by giving a high-fat diet over a long period of time is suppressed. It became clear that it was possible.
- Example 3 ⁇ Effect of flurbiprofen on the rise in blood leptin concentration> (Method for increasing blood leptin concentration and method for confirming blood leptin concentration)
- a normal diet (10 kcal% fat, D12450B, Research DIET) or a high fat diet (60 kcal% fat, D12492, Research DIET).
- Flurbiprofen (F8514, manufactured by SIGMA) was orally administered as drinking water by diluting with drinking water to a dose of about 10 mg / kg per day.
- mice were divided into 4 groups of 7 to 8 animals, and the following meals were given to the mice of each group.
- Group 1 Normal diet (10 kcal% fat) only
- Group 2 High fat diet (60 kcal% fat) only
- Group 3 Normal diet + flurbiprofen
- Group 4 High fat diet + flurbiprofen High fat diet or normal diet
- From the 8 week-old mice 20-25 ⁇ L of blood was collected from the tail before or after fasting for 28-29.5 hours. The collected blood was added with 1 ⁇ L of 0.5 M EDTA and centrifuged (3000 rpm, 15 minutes). Using the supernatant plasma obtained by centrifugation, the leptin concentration in plasma was measured by ELISA.
- FIG. 6 is a graph showing the results of measuring the leptin concentration in plasma before and after fasting.
- 6A shows the leptin concentration in plasma before fasting
- FIG. 6B shows the leptin concentration in plasma after fasting.
- the values shown in the graph of FIG. 6 (b) show the leptin concentration in the plasma after fasting in each group of mice as the leptin concentration in the plasma after fasting in each mouse ⁇ standard deviation.
- FIG. 7 is a graph showing the correlation between leptin concentration in plasma and body weight.
- FIG. 7A shows the correlation between leptin concentration in plasma before fasting and body weight
- FIG. 7B shows the correlation between leptin concentration in plasma after fasting and body weight.
- “R” shown in FIG. 7 represents a correlation coefficient.
- group 2 mice fed a high fat diet for 8 weeks had plasma leptin concentrations compared to group 1 mice fed a normal diet. Significantly higher.
- the leptin concentration in plasma was significantly lower in the mice of group 4 administered with the high-fat diet + flurbiprofen for 8 weeks compared with the mice of group 2 fed with the high-fat diet.
- FIG. 7 there was a correlation between the body weight of the mouse and the leptin concentration in plasma before and after fasting. From these results, it was clarified that an increase in plasma leptin concentration can be suppressed by simultaneously administering a high fat diet and flurbiprofen.
- leptin does not function in individuals in which leptin resistance is induced, a mechanism for maintaining homeostasis of the living body works and more leptin is secreted. As a result, the leptin level in blood rises in individuals in which leptin resistance is induced.
- leptin is functioning normally by preventing leptin resistance, so that it is not necessary to secrete more leptin. For this reason, it was considered that the blood leptin level was maintained at a normal level in the mice of Group 4 administered with the high fat diet + flurbiprofen for 8 weeks.
- Example 4 ⁇ Blood glucose reduction effect of flurbiprofen> It has been reported that leptin resistance is involved in the development of diabetes (eg, Gwo-Hwa Lee et al, Abnormal splicing of the leptin receptor in diabetic mice, Nature 1996, 379 (15): 632-635; Hong Chen et al, Evidence That the Diabetes Gene Encodes the Leptin Receptor: Identification of a Mutation in the Leptin Receptor Gene in db / db Mice, Cell 1996, 84: 491-495). Specifically, db / db mice that have mutations in the leptin high affinity receptor (OB-R) and are defective in leptin signaling develop diabetes. For this reason, it is considered that it is possible to confirm whether or not leptin resistance has been induced using a change in blood glucose level in the glucose tolerance test as one index.
- OB-R leptin high affinity receptor
- Example 4 it was examined whether the induction of leptin resistance could be evaluated using the change in blood glucose level in the glucose tolerance test as one index. At the same time, using the change in blood glucose level in the glucose tolerance test as one index, whether or not induction of leptin resistance can be prevented by administration of flurbiprofen was examined. Specifically, 14-week-old male mice (C57BL / 6 Cr Slc) were divided into four groups of 14 to 16, and the following meals were given to the mice of each group for 8 weeks, as in Example 3. It was.
- Group 1 Normal diet (10 kcal% fat) only
- Group 2 High fat diet (HFD) (60 kcal% fat) only
- Group 3 Normal diet + flurbiprofen
- Group 4 High fat diet + flurbipro Fen Note that flurbiprofen (F8514, manufactured by SIGMA) was orally administered as drinking water by diluting with drinking water to a dose of 10 mg / kg per day.
- a glucose tolerance test was performed on mice of each group 8 weeks after feeding a high fat diet or a normal diet.
- the glucose tolerance test is the most basic method for evaluating glucose tolerance and insulin secretion ability.
- the mice were fasted for 17 hours from the night before the glucose tolerance test, and the fasting blood glucose level (pre) was measured from 10:00 am on the next day using Freestyle freedom (Niplo Blood Glucose Self-Measurement System). Thereafter, glucose was administered intraperitoneally (ip) to a dose of 2 g / kg, and blood glucose levels after 30 minutes, 1 hour and 2 hours were measured with Freestyle freedom (Niplo Blood Glucose Self-Measurement System). did.
- FIG. 8 is a graph showing the results of the glucose tolerance test.
- Statistical analysis was performed using t-test. Note that “*” in FIG. 8 indicates a risk rate of 5% between the blood glucose level of the group 2 mice fed with a high fat diet for 8 weeks and the blood glucose level of the group 1 mice fed a normal diet. Indicates that there is a significant difference, and “***” indicates that there is a significant difference when the risk rate is less than 0.1%.
- “#” in FIG. 8 indicates between the blood glucose level in the group 4 mice administered with the high fat diet + flurbiprofen for 8 weeks and the blood glucose level in the group 2 mice fed with the high fat diet. Indicates that there is a significant difference when the risk rate is less than 5%, “##” indicates that there is a significant difference when the risk rate is less than 1%, and “##” indicates that the risk rate is less than 0.1%. It shows that there is a significant difference.
- the group 2 mice fed with a high fat diet had significantly higher blood glucose levels 2 hours after administration of glucose than the mice of group 1 fed with a normal diet.
- the results shown in FIG. It shows that it can also be evaluated by confirming a change in blood glucose level in the test.
- the blood glucose level of the mice of Group 4 rapidly decreased 1 hour after glucose administration.
- a significant difference was observed between the blood glucose level of group 2 and the blood glucose level of group 4 1 hour after glucose administration. From this, it was confirmed that the induction of leptin resistance was prevented by administration of flurbiprofen, using the change in blood glucose level in the glucose tolerance test as an index.
- Example 5 ⁇ Effects of high fat diet and flurbiprofen administration on mouse fat mass>
- mice C57BL / 6 Cr Slc
- Group 1 Normal diet (10 kcal% fat) only
- Group 2 High fat diet (60 kcal% fat) only
- Group 3 Normal diet + flurbiprofen
- Group 4 High fat diet + flurbiprofen Flurbiprofen (F8514, manufactured by SIGMA) was orally administered as drinking water by diluting with drinking water so as to have a dose of 10 mg / kg per day.
- mice fed with a high fat diet or a normal diet for 8 weeks were dissected along the midline, and the amount of adipose tissue (built-in fat) was measured. Built-in fat was extracted from the abdomen of the mouse and the weight was measured.
- FIG. 9 is a graph showing the amount of fat in each group of mice at 8 weeks after administration of flurbiprofen.
- Statistical analysis was performed using t-test. Note that “***” in FIG. 9 is a risk rate between the fat mass in the group 2 mice fed the high fat diet for 8 weeks and the fat mass in the group 1 mice fed the normal diet. It shows that there is a significant difference at less than 0.1%.
- “#” indicates a risk factor of 5 between the amount of fat in the mice of group 4 administered with the high fat diet + flurbiprofen for 8 weeks and the amount of fat in the mice of group 2 fed with the high fat diet. It shows that there is a significant difference in less than%.
- Example 6 ⁇ Effects of high-fat diet and flurbiprofen on mouse activity> In order to measure the amount of behavior of the mouse, an Open Field Test was performed. Specifically, 4 weeks old male mice (C57BL / 6 Cr Slc) were divided into 4 groups of 15 to 16 mice, and as in Example 5, the following meals were given to each group of mice for 8 weeks. Gave.
- Group 1 Normal diet (10 kcal% fat) only
- Group 2 High fat diet (60 kcal% fat) only
- Group 3 Normal diet + flurbiprofen
- Group 4 High fat diet + flurbiprofen Flurbiprofen (F8514, manufactured by SIGMA) was orally administered as drinking water by diluting with drinking water so as to have a dose of 10 mg / kg per day.
- the Open Field Test is a method in which a mouse is placed in the center of a square field (48 ⁇ 48 cm) and the amount of movement of the mouse in a new environment is digitized and measured. Specifically, using a spontaneous movement measurement device (SCANET MV-10MT, manufactured by Toyo Sangyo Co., Ltd.), between 16:30 and 17:30 before the feeding behavior of mice was activated, The amount of action for 5 minutes was measured.
- SCANET MV-10MT spontaneous movement measurement device
- FIG. 10 is a graph showing the amount of behavior of each group of mice at 8 weeks after administration of flurbiprofen.
- Statistical analysis was performed using t-test.
- Example 7 Prevention of leptin resistance by flurbiprofen-Examination of food intake-> When leptin resistance is induced, although the leptin concentration in the blood is increased, the feeding inhibitory action and the energy consumption enhancing action by leptin are reduced. From this, the presence or absence of leptin resistance can be confirmed using the presence or absence of an antifeeding effect of leptin as an index. Therefore, in Example 7, it was further confirmed that the induction of leptin resistance was prevented by flurbiprofen using the presence or absence of an antifeeding effect by leptin as an index.
- mice 4 weeks old male mice (C57BL / 6 Cr Slc) were divided into 4 groups of 6 to 8 animals, and the mice of each group were given the following meals for 8 weeks as in Example 5.
- FIG. 11 is a diagram showing the procedure of the experiment of Example 7.
- the day of isolation of mice that had been fed with a high-fat diet or a normal diet for 8 weeks into 1 mouse / cage was defined as day 0 (Day 0), and acclimated for 3 days ( habituation).
- the abdominal cavity of saline (saline) was adjusted to 5 mL / kg as a dummy on the third day (Day 3) and the fourth day (Day 4). Internal (ip) administration was performed.
- physiological saline was intraperitoneally (ip) administered at 5 mL / kg on the 5th day (Day 5), and the amount of food intake up to 24 hours after the physiological saline administration was controlled on the 6th day (Day 6). It was.
- leptin model number: 498-OB, rmeptin, manufactured by R & D
- ip intraperitoneally
- 7th day Day 7
- the amount of food intake up to 24 hours after leptin administration was measured.
- the effect of leptin on food intake using the ratio of food intake on Day 6 to Day 7 on Day 5 (Day 5) to Day 6 (Day 6) (Change in food intake) was examined.
- the rate of change in food intake is expressed by the following formula (1).
- Rate of change in food intake (food intake from the 6th to 7th days / food intake from the 5th to 6th days) ⁇ 100 (1) All administrations of leptin and physiological saline were performed twice a day at regular intervals (9:00 and 19:00).
- FIG. 12 is a graph showing changes in food intake of mice in each group after leptin administration.
- Statistical analysis was performed using t-test. Note that “*” in FIG. 12 is between the rate of change in the amount of intake in the mice of group 2 given the high fat diet for 8 weeks and the rate of change in the amount of intake in the mice of group 1 given the normal diet. It shows that there is a significant difference when the risk rate is less than 5%.
- mice of group 2 fed with the high fat diet for 8 weeks the feeding suppression by leptin did not occur and the amount of food intake was rather increased.
- group 4 which was administered a high fat diet + flurbiprofen for 8 weeks the administration of flurbiprofen suppressed feeding by leptin.
- group 4 mice administered high-fat diet plus flurbiprofen for 8 weeks There was a significant difference between the rate of change in food intake in group 4 mice administered high-fat diet plus flurbiprofen for 8 weeks and the rate of change in food intake in group 1 mice fed a normal diet.
- mice administered with a high fat diet plus flurbiprofen have no difference in the rate of change in food intake compared to mice administered with a normal diet. Therefore, it is considered that leptin resistance was prevented in mice administered with a high fat diet + flurbiprofen.
- flurbiprofen is useful for preventing leptin resistance.
- Example 8 ⁇ Weight loss effect of flurbiprofen (therapeutic effect)>
- flurbiprofen was also useful in improving leptin resistance, using changes in body weight of mice as an index. Specifically, 4-week-old male mice (C57BL / 6 Cr Slc) were fed with a high fat diet (High Fat Diet; HFD) for 6 months to induce leptin resistance. These mice were then divided into two groups (14 each), and each group of mice was fed the following diet for 3 weeks.
- a high fat diet High Fat Diet; HFD
- Control group normal diet (10 kcal% fat) only flurbiprofen administration group (Ful): normal diet + flurbiprofen Flurbiprofen (F8514, manufactured by SIGMA) is 3 mg / day It was diluted with drinking water to a dose of kg and orally administered as drinking water.
- the body weight change rate of each group of mice was measured based on the start date (week 0) of flurbiprofen administration. In addition, the amount of internal fat (g) at 3 weeks was measured. Built-in fat was extracted from the abdomen of the mouse and the weight was measured.
- FIG. 13 is a graph showing the weight change rate of mice in the control group and the flurbiprofen administration group.
- Statistical analysis was performed using t-test. Note that “*” in FIG. 13 indicates that there is a significant difference between the weight change rate in the control group mice and the body weight change rate in the flurbiprofen-administered group mice at a risk rate of less than 5%. ing.
- FIG. 14 is a graph showing the amount of internal fat (g) after 3 weeks of normal diet in mice in the control group and flurbiprofen administration group.
- Statistical analysis was performed using t-test. Note that “**” in FIG. 14 indicates that there is a significant difference between the built-in fat mass in the mice in the control group and the built-in fat mass in the mice in the flurbiprofen administration group at a risk rate of less than 1%. Show.
- the average body weight before administration of flurbiprofen was 53.879 ⁇ 0.6299 g in the control group and 53.479 ⁇ 0.7804 g in the flurbiprofen administration group.
- the mean body weight after 3 weeks of flurbiprofen administration was 48.371 ⁇ 0.6387 g in the control group and 45.543 ⁇ 1.1103 g in the flurbiprofen administration group. It was observed (P ⁇ 0.05). This result indicates that leptin resistance can be improved by administering a clinical level of dosage (3 mg / kg per day) of flurbiprofen. Further, it was confirmed that the effect of improving the leptin resistance by flurbiprofen appears one week after administration of flurbiprofen.
- Example 9 ⁇ Reduction of blood leptin concentration by administration of flurbiprofen>
- leptin resistance was induced by feeding male mice (C57BL / 6 Cr Slc) 4 weeks old with a high fat diet (HFD) for 6 months. These mice were then divided into two groups (8 mice each), and each group of mice was fed the following diet for 3 weeks.
- FIG. 15 shows the results of measurement of blood leptin concentration 3 weeks after administration of flurbiprofen.
- the blood leptin concentration of the mice in the control group was 106.51 ⁇ 3.7122 (ng / ml).
- the blood leptin concentration of the mice in the flurbiprofen administration group is 87.105 ⁇ 5.4415 (ng / ml), and the blood leptin concentration is significantly reduced by administration of flurbiprofen. (P ⁇ 0.05).
- Example 10 ⁇ Change in body length by administration of flurbipropene>
- leptin resistance was induced by feeding male mice (C57BL / 6 Cr Slc) 4 weeks old with a high fat diet (HFD) for 6 months. These mice were then divided into two groups (8 mice each), and each group of mice was fed the following diet for 3 weeks.
- FIG. 16 shows the results of measuring the length of mice 3 weeks after administration of flurbiprofen.
- Example 11 ⁇ Effect of flurbiprofen on blood glucose improvement>
- leptin resistance was induced by feeding male mice (C57BL / 6 Cr Slc) 4 weeks old with a high fat diet (HFD) for 6 months. These mice were then divided into two groups (8 mice each), and each group of mice was fed the following diet for 3 weeks.
- Control group normal diet (10 kcal% fat)
- flurbiprofen administration group normal diet + flurbiprofen Flurbiprofen (F8514, manufactured by SIGMA) has a dose of 3 mg / kg per day As described above, it was diluted with drinking water and orally administered as drinking water.
- a glucose tolerance test was performed on mice 3 weeks after administration of flurbiprofen.
- Example 11 (Glucose tolerance test) In Example 11, the mice were fasted for 17 hours from the night before (17:00 to 18:00) of the glucose tolerance test, and the fasting blood glucose level (pre) was measured from the next morning (10:00 to 11:00) to Freestyle freedom (Nipro Corporation). Company blood glucose self-measurement system). Thereafter, glucose was administered intraperitoneally (ip) to a dose of 2 g / kg, and blood glucose levels at 30 minutes, 1 hour, 2 hours, 3 hours and 4 hours were measured according to Freestyle freedom (Nipro Corporation). Company blood glucose self-measurement system).
- ip intraperitoneally
- FIG. 17 shows the result of measuring the blood glucose level.
- FIG. 17 is a graph showing the results of a glucose tolerance test.
- the blood glucose level at each time after glucose administration is expressed as a percentage (%) when the fasting blood glucose level is taken as 100%.
- Statistical analysis was performed using t-test.
- flurbiprofen is also useful in improving leptin resistance.
- Example 12 ⁇ Effects of high-fat diet and flurbiprofen on the amount of water consumed by mice>
- male mice C57BL / 6 Cr Slc
- 4 weeks old were divided into 4 groups of 7 to 8 mice, and the mice were isolated so as to have one cage. And acclimated for 2 days.
- the amount of drinking water was measured every 24 hours from the morning of the second day of acclimatization, and the total amount for 4 days was totaled.
- Group 1 Normal diet (10 kcal% fat) only
- Group 2 High fat diet (60 kcal% fat) only
- Group 3 Normal diet + flurbiprofen
- Group 4 High fat diet + flurbiprofen Flurbiprofen (F8514, manufactured by SIGMA) was orally administered as drinking water by diluting with drinking water so as to have a dose of 10 mg / kg per day.
- FIG. 18 is a graph showing the result of measuring the amount of drinking water.
- Statistical analysis was performed using t-test.
- the leptin resistance can be improved and / or prevented by using the pharmaceutical composition according to the present invention. Furthermore, when the pharmaceutical composition according to the present invention is used, it is effective in improving and / or preventing related diseases associated with leptin resistance, such as diabetes, hypertension, hyperlipidemia, arteriosclerosis and the like. Conceivable. Therefore, the invention according to the present invention can be used in the pharmaceutical industry.
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Abstract
Description
本発明にかかる薬学的組成物は、レプチン抵抗性を改善および/または予防するための薬学的組成物であって、フルルビプロフェンを含有することを特徴としている。本明細書において、「フルルビプロフェン」は、以下の一般式で表される化合物(I)およびその鏡像異性体、並びにこれらの薬学的に許容される塩が包含される。
なお、本明細書中において、上記「非経口」とは、脳室内、静脈内、筋肉内、腹腔内、胸骨内、皮下、および関節内の注射および注入を含む投与の様式をいう。
<レプチン抵抗性に対するフルルビプロフェンの効果1>
(レプチン抵抗性の誘発方法および確認方法)
まず、本実施例で用いたレプチン抵抗性の誘発方法およびレプチン抵抗性の確認方法について以下に説明する。
マウスを7~8匹ずつ計4つのグループに分け、各グループのマウスには以下の食事を与えた。
グループ1:普通食(10kcal%fat、D12450B、Reserch DIET社製)のみ
グループ2:高脂肪食(60kcal%fat、D12492、Reserch DIET社製)のみ
グループ3:普通食+フルルビプロフェン(F8514、SIGMA社製)
グループ4:高脂肪食+フルルビプロフェン(F8514、SIGMA社製)
なお、フルルビプロフェンは、1日当たりおよそ10mg/kgの投与量になるように飲用水を用いて希釈して、飲用水として経口投与した。その後、7日ごとにマウスの体重を測定した。
結果を図3に示す。図3は、フルルビプロフェン投与後8週間における各グループのマウスの体重の変化を示すグラフである。図3のグラフに示す値は、各グループにおけるマウスの体重を、それぞれのマウスの体重の平均値±標準偏差として示した(グループあたりn=7~8)。また、t検定を用いて統計解析を行った。なお、グラフ中に示す「*」は、危険率5%未満において、グループ2の体重とグループ4の体重との間に有意差があることを表す。
<レプチン抵抗性に対するフルルビプロフェンの効果2>
(レプチン抵抗性の誘発方法および確認方法)
4週齢の雄のマウス(C57BL/6 Cr Slc)に普通食(10kcal%fat、D12450B、Reserch DIET社製)または高脂肪食(60kcal%fat、D12492、Reserch DIET社製)を与えた。フルルビプロフェンは、1日当たりおよそ10mg/kgの投与量になるように飲用水を用いて希釈して、飲用水として経口投与した。
グループ1:普通食(10kcal%fat)のみ
グループ2:高脂肪食(60kcal%fat)のみ
グループ3:普通食+フルルビプロフェン
グループ4:高脂肪食+フルルビプロフェン
高脂肪食または普通食を与えて8週目のマウスに、マウス由来のレプチン(450-31、PeproTech社製または498-OB、rmLeptin、R&D社製)を生理食塩水に溶解して調製したものを、投与量が1mg/kgになるように、非経口的に尾静脈内投与した。レプチン投与のコントロールとしては、等量の生理食塩水を尾静脈内投与した。
<血中レプチン濃度の上昇に対するフルルビプロフェンの効果>
(血中レプチン濃度を上昇させる方法および血中レプチン濃度の確認方法)
4週齢の雄のマウス(C57BL/6 Cr Slc)に普通食(10kcal%fat、D12450B、Reserch DIET社製)または高脂肪食(60kcal%fat、D12492、Reserch DIET社製)を与えた。フルルビプロフェン(F8514、SIGMA社製)は、1日当たりおよそ10mg/kgの投与量になるように飲用水を用いて希釈して、飲用水として経口投与した。
グループ1:普通食(10kcal%fat)のみ
グループ2:高脂肪食(60kcal%fat)のみ
グループ3:普通食+フルルビプロフェン
グループ4:高脂肪食+フルルビプロフェン
高脂肪食または普通食を与えて8週目のマウスから、絶食前または28~29.5時間絶食後の血液をそれぞれ20~25μL、尾部から採取した。採取した血液は、0.5M EDTAを1μL加えて遠心分離(3000rpm、15分)を行った。遠心分離によって得られた上清の血漿を用いて、ELISA法により血漿中のレプチン濃度を測定した。
<フルルビプロフェンによる血糖値減少効果>
レプチン抵抗性が糖尿病の発症に関与することが報告されている(例えば、Gwo-Hwa Lee et al, Abnormal splicing of the leptin receptor in diabetic mice, Nature 1996, 379(15):632-635; Hong Chen et al, Evidence That the Diabetes Gene Encodes the Leptin Receptor: Identification of a Mutation in the Leptin Receptor Gene in db/db Mice, Cell 1996, 84: 491-495等を参照)。具体的には、レプチン高親和性受容体(OB-R)に変異を有し、レプチンシグナル伝達に欠陥があるdb/dbマウスは、糖尿病を発症する。このため、グルコース負荷試験における血糖値の変化を1つの指標として、レプチン抵抗性が誘発されているか否かを確認することができると考えられる。
グループ1:普通食(10kcal%fat)のみ
グループ2:高脂肪食(High Fat Diet:HFD)(60kcal%fat)のみ
グループ3:普通食+フルルビプロフェン
グループ4:高脂肪食+フルルビプロフェン
なお、フルルビプロフェン(F8514、SIGMA社製)は、1日当たり10mg/kgの投与量になるように飲用水を用いて希釈して、飲用水として経口投与した。
グルコース負荷試験は、耐糖能とインスリン分泌能を評価する最も基本的な方法である。実施例4では、グルコース負荷試験の前日の夜からマウスを17時間絶食させ、翌日午前10時から空腹時血糖値(pre)をFreestyle freedom(ニプロ株式会社 血糖自己測定システム)によって測定した。その後、2g/kgの投与量になるようにグルコースを腹腔内(i.p.)投与し、30分後、1時間後および2時間後の血糖値をFreestyle freedom(ニプロ株式会社 血糖自己測定システム)によって測定した。
<高脂肪食およびフルルビプロフェン投与によるマウス脂肪量への影響>
4週齢の雄マウス(C57BL/6 Cr Slc)を7~8匹ずつ計4つのグループに分け、実施例4と同様に、各グループのマウスには以下の食事を8週間与えた。
グループ1:普通食(10kcal%fat)のみ
グループ2:高脂肪食(60kcal%fat)のみ
グループ3:普通食+フルルビプロフェン
グループ4:高脂肪食+フルルビプロフェン
なお、フルルビプロフェン(F8514、SIGMA社製)は、1日当たり10mg/kgの投与量になるように飲用水を用いて希釈して、飲用水として経口投与した。
<マウス行動量に対する高脂肪食、フルルビプロフェンの影響>
マウスの行動量を測定するために、Open Field Testを行った。具体的には、4週齢の雄マウス(C57BL/6 Cr Slc)を15~16匹ずつ計4つのグループに分け、実施例5と同様に、各グループのマウスには以下の食事を8週間与えた。
グループ1:普通食(10kcal%fat)のみ
グループ2:高脂肪食(60kcal%fat)のみ
グループ3:普通食+フルルビプロフェン
グループ4:高脂肪食+フルルビプロフェン
なお、フルルビプロフェン(F8514、SIGMA社製)は、1日当たり10mg/kgの投与量になるように飲用水を用いて希釈して、飲用水として経口投与した。
Open Field Testは、マウスを正方形のフィールド(48×48cm)の中央に静置し、新規環境でのマウスの行動量を数値化して測定する方法である。具体的には、自発運動測定装置(SCANET MV-10MT,東洋産業株式会社製)を用いて、マウスの摂食行動が活発化する前の16:30~17:30の間に、一匹あたり5分間の行動量を測定した。
<フルルビプロフェンによるレプチン抵抗性の予防~摂食量の検討~>
レプチン抵抗性が誘発されると、血中レプチン濃度が上昇するにもかかわらず、レプチンによる摂食抑制作用やエネルギー消費亢進作用が低下する。このことから、レプチンによる摂食抑制作用の有無を指標として、レプチン抵抗性の有無を確認することができる。そこで、実施例7では、レプチンによる摂食抑制作用の有無を指標として、フルルビプロフェンによってレプチン抵抗性の誘発が予防されることをさらに確認した。
グループ1:普通食(10kcal%fat)のみ
グループ2:高脂肪食(60kcal%fat)のみ
グループ3:普通食+フルルビプロフェン
グループ4:高脂肪食+フルルビプロフェン
なお、フルルビプロフェン(F8514、SIGMA社製)は、1日当たり10mg/kgの投与量になるように飲用水を用いて希釈して、飲用水として経口投与した。
摂食量変化率(%)=(6日目~7日目の摂食量/5日目~6日目の摂食量)×100 … (1)
なお、レプチンおよび生理食塩水の投与は全て、1日2回、定刻(9:00および19:00)に行った。
<フルルビプロフェンの体重減少効果(治療効果)>
実施例8では、レプチン抵抗性の改善においてもフルルビプロフェンが有用であることを、マウスの体重の変化を指標として確認した。具体的には、4週齢の雄マウス(C57BL/6 Cr Slc)を高脂肪食(High Fat Diet;HFD)を6ヶ月間与えてレプチン抵抗性を誘発した。その後、これらのマウスを2つのグループ(各14匹ずつ)に分け、各グループのマウスには以下の食事を3週間与えた。
コントロール群(Cont):普通食(10kcal%fat)のみ
フルルビプロフェン投与群(Ful):普通食+フルルビプロフェン
なお、フルルビプロフェン(F8514、SIGMA社製)は、1日当たり3mg/kgの投与量になるように飲用水を用いて希釈して、飲用水として経口投与した。
<フルルビプロフェン投与による血中レプチン濃度の減少>
実施例8と同様に、4週齢の雄マウス(C57BL/6 Cr Slc)を高脂肪食(High Fat Diet;HFD)を6ヶ月間与えてレプチン抵抗性を誘発した。その後、これらのマウスを2つのグループ(各8匹ずつ)に分け、各グループのマウスには以下の食事を3週間与えた。
コントロール群(Cont):普通食(10kcal%fat)のみ
フルルビプロフェン投与群(Ful):普通食+フルルビプロフェン
なお、フルルビプロフェン(F8514、SIGMA社製)は、1日当たり3mg/kgの投与量になるように飲用水を用いて希釈して、飲用水として経口投与した。
<フルルビプロプェン投与による体長の変化>
実施例8と同様に、4週齢の雄マウス(C57BL/6 Cr Slc)を高脂肪食(High Fat Diet;HFD)を6ヶ月間与えてレプチン抵抗性を誘発した。その後、これらのマウスを2つのグループ(各8匹ずつ)に分け、各グループのマウスには以下の食事を3週間与えた。
コントロール群(Cont):普通食(10kcal%fat)のみ
フルルビプロフェン投与群(Ful):普通食+フルルビプロフェン
なお、フルルビプロフェン(F8514、SIGMA社製)は、1日当たり3mg/kgの投与量になるように飲用水を用いて希釈して、飲用水として経口投与した。
<フルルビプロフェンによる血糖値改善効果>
実施例8と同様に、4週齢の雄マウス(C57BL/6 Cr Slc)を高脂肪食(High Fat Diet;HFD)を6ヶ月間与えてレプチン抵抗性を誘発した。その後、これらのマウスを2つのグループ(各8匹ずつ)に分け、各グループのマウスには以下の食事を3週間与えた。
コントロール群:普通食(10kcal%fat)のみ
フルルビプロフェン投与群:普通食+フルルビプロフェン
なお、フルルビプロフェン(F8514、SIGMA社製)は、1日当たり3mg/kgの投与量になるように飲用水を用いて希釈して、飲用水として経口投与した。
実施例11では、グルコース負荷試験の前日の夜(17時から18時)からマウスを17時間絶食させ、翌日午前(10時から11時)から空腹時血糖値(pre)をFreestyle freedom(ニプロ株式会社 血糖自己測定システム)によって測定した。その後、2g/kgの投与量になるようにグルコースを腹腔内(i.p.)投与し、30分後、1時間後、2時間後、3時間後および4時間後の血糖値をFreestyle freedom(ニプロ株式会社 血糖自己測定システム)によって測定した。
<マウスの飲水量に対する高脂肪食、フルルビプロフェンの影響>
マウスの飲水量を測定するために、4週齢の雄マウス(C57BL/6 Cr Slc)を7~8匹ずつ計4つのグループに分け、マウスを1ケージ1匹となるように隔離(isolation)し、2日間馴化させた。馴化2日目の午前から24時間おきに4日間飲水量を測定し、4日間の合計量を集計した。
グループ1:普通食(10kcal%fat)のみ
グループ2:高脂肪食(60kcal%fat)のみ
グループ3:普通食+フルルビプロフェン
グループ4:高脂肪食+フルルビプロフェン
なお、フルルビプロフェン(F8514、SIGMA社製)は、1日当たり10mg/kgの投与量になるように飲用水を用いて希釈して、飲用水として経口投与した。
Claims (2)
- フルルビプロフェンを含有することを特徴とする、レプチン抵抗性を改善および/または予防するための薬学的組成物。
- レプチン抵抗性を改善および/または予防するための薬学的組成物を製造するための、フルルビプロフェンの使用。
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US13/263,966 US9132108B2 (en) | 2009-04-17 | 2010-04-14 | Pharmaceutical composition for ameliorating and/or preventing leptin resistance, and use thereof |
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JP2015078145A (ja) * | 2013-10-16 | 2015-04-23 | 国立大学法人広島大学 | アルデヒドデヒドロゲナーゼの活性を増強するための医薬組成物 |
JP2015531376A (ja) * | 2012-09-27 | 2015-11-02 | ザ チルドレンズ メディカル センター コーポレイション | 肥満の処置のための化合物およびそれの使用方法 |
JP2017513824A (ja) * | 2014-03-26 | 2017-06-01 | ザ チルドレンズ メディカル センター コーポレイション | 肥満の処置のためのセラストロールおよび誘導体 |
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JP2008208091A (ja) * | 2007-02-27 | 2008-09-11 | Hiroshima Univ | 小胞体ストレス関連疾患の治療薬および予防薬 |
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WO2004078113A2 (en) | 2003-03-04 | 2004-09-16 | Pharmacia Corporation | Treatment and prevention of obesity with cox-2 inhibitors alone or in combination with weight-loss agents |
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Non-Patent Citations (2)
Title |
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LALE OZCAN ET AL.: "Endoplasmic Reticulum Stress Plays a Central Role in Development of Leptin Resistance", CELL METABOLISM, vol. 9, 7 January 2009 (2009-01-07), pages 35 - 51 * |
TORU HOSOI ET AL.: "Endoplasmic Reticulum Stress Induces Leptin Resistance", MOLECULAR PHARMACOLOGY, vol. 74, no. 6, 2008, pages 1610 - 1619 * |
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JP2015531376A (ja) * | 2012-09-27 | 2015-11-02 | ザ チルドレンズ メディカル センター コーポレイション | 肥満の処置のための化合物およびそれの使用方法 |
JP2019070052A (ja) * | 2012-09-27 | 2019-05-09 | ザ チルドレンズ メディカル センター コーポレーション | 肥満の処置のための化合物およびそれの使用方法 |
JP2015078145A (ja) * | 2013-10-16 | 2015-04-23 | 国立大学法人広島大学 | アルデヒドデヒドロゲナーゼの活性を増強するための医薬組成物 |
JP2017513824A (ja) * | 2014-03-26 | 2017-06-01 | ザ チルドレンズ メディカル センター コーポレイション | 肥満の処置のためのセラストロールおよび誘導体 |
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US9132108B2 (en) | 2015-09-15 |
JPWO2010119674A1 (ja) | 2012-10-22 |
JP5780549B2 (ja) | 2015-09-16 |
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