JP2006327959A - Remedy for endocrine disease and metabolic disease - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a remedy for endocrine disease, and to provide a remedy for metabolic disease. <P>SOLUTION: The remedies each contain a hyaluronan as an active ingredient. The hyaluronan preferably comprises a tetrasaccharide (HA4) composed of two disaccharide units of which the each comprises a -D-glucuronic acid-β-1,3-D-N-acetylglucosamine-β-1,4- unit. <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

  The present invention relates to an endocrine disease therapeutic agent and a metabolic disease therapeutic agent containing hyaluronan as an active ingredient.

  Hyaluronan is a long-chain polysaccharide composed of disaccharide repeating units of D-glucuronic acid and N-acetyl-D-glucosamine, while oligosaccharides are also known. Hyaluronan is produced by extraction from biological tissues such as chicken crowns, corpses, skin, joint fluids, or fermentation methods using bacteria of the genus Streptococcus, and has no toxicological or immunological effects. For example, treatment of arthritis by intraarticular injection of hyaluronan is well known. In the following description, the tetrasaccharide hyaluronan is referred to as HA4.

  HA4 has been reported to have therapeutic / suppressive effects on organ preservation, liver damage, and gastric ulcers (see Patent Document 1). Moreover, it is known that HA4 has a stress protein expression enhancing action and a cell death suppressing action (see Non-Patent Document 1).

  On the other hand, diabetes is one of lifestyle-related diseases, and is known as a disease in which blood sugar level is high (hyperglycemia) and glucose is also detected in urine. If diabetes is left unattended because it has no subjective symptoms, hyperglycemia can cause damage to various organs throughout the body. In particular, organs such as nerves and blood vessels are easily affected, and when they progress, complications such as neuropathy, nephropathy, and retinopathy occur.

WO2002 / 004471 Xu H, Ito T, Tawada A, Maeda H, Yamanokuchi H, Isahara K, Yoshida K, Uchiyama Y, Asari A. Effect of hyaluronan oligosaccharides on the expression of heat shock protein 72. J Biol Chem. 2002 10; 277 (19 ): 17308-14.

  An object of the present invention is to provide an endocrine disease therapeutic agent and a metabolic disease therapeutic agent.

  The therapeutic agent for endocrine disease and the therapeutic agent for metabolic disease according to the present invention that have achieved the above-mentioned object contain hyaluronan as an active ingredient.

  Since the drug according to the present invention contains hyaluronan as an active ingredient, it has an advantage that it can be easily mass-produced at a relatively low cost. Hyaluronan is expected to be a therapeutic agent with extremely few side effects because it has little toxicity and antigenicity, and enhances the therapeutic action and disease prevention action inherent in the living body. Thus, according to the present invention, a novel drug effective for endocrine diseases and metabolic diseases can be provided.

  Hereinafter, the present invention will be described in detail. In the following description, the therapeutic agent according to the present invention is simply referred to as a drug.

  The hyaluronan contained in the drug according to the present invention basically includes at least one disaccharide unit in which the 1-position of β-D-glucuronic acid and the 3-position of β-D-N-acetylglucosamine are bonded. As long as it is more than sugar and is basically composed of β-D-glucuronic acid and β-DN-acetylglucosamine, those elements are combined with one or more disaccharide units combined. May be used, and derivatives thereof such as those having a hydrolyzable protecting group such as an acyl group can also be used. The sugar may be an unsaturated sugar, and examples of the unsaturated sugar include non-reducing terminal sugars, usually those in which the 4- and 5-position carbons of glucuronic acid are unsaturated. As the hyaluronan used in the present invention, specifically, those extracted from natural products such as animals, those obtained by culturing microorganisms, those synthesized chemically or enzymatically, etc. should be used. Can do. For example, it can be obtained by known extraction and purification methods from biological tissues such as chicken crown, corpse, skin, and joint fluid. It can also be produced by a fermentation method using Streptococcus bacteria or the like.

  In the present invention, hyaluronan oligosaccharides are also included in hyaluronan, and use from low molecular weight hyaluronan such as the above-mentioned disaccharide consisting of one disaccharide unit and its derivatives to high molecular weight hyaluronan having a weight average molecular weight of about 4 million. be able to. Preferably, hyaluronan having a weight average molecular weight of about 380 to 900,000, which is excellent in terms of permeability in tissues, and the like, more preferably about 2 to 20 sugars.

  Specifically, hyaluronan having a low molecular weight can be obtained by known methods such as enzymatic decomposition, alkaline decomposition, heat treatment, and ultrasonic treatment (Biochem., 33 (1994) p6503-6507). It is preferably produced by a method of reducing the molecular weight, a method of chemically or enzymatically synthesizing (Glycoconjugate J., (1993) p435-439, WO93 / 20827). For example, hyaluronan degrading enzymes (hyaluronidase (derived from testicles), hyaluronidase (derived from Streptomyces), hyaluronidase SD, etc.), chondroitinase AC, chondroitinase ACII, chondroitinase ACIII, chondroitinase ABC, etc. A method to produce hyaluronan oligosaccharides by causing an enzyme that degrades hyaluronan to produce hyaluronan oligosaccharides (see Shinsei Chemistry Laboratory "Carbohydrate II-Proteoglycans and Glycosaminoglycans" p244-248, published in 1991, Tokyo Chemical Dojin) Can be mentioned.

  As an alkali decomposition method, for example, a base such as sodium hydroxide of about 1N is added to a hyaluronan solution, heated for several hours to lower the molecular weight, and then neutralized by adding an acid such as hydrochloric acid. And a method for obtaining a low molecular weight hyaluronan. Hyaluronan used in the present invention includes a salt form, and a pharmaceutically acceptable salt thereof can be used as necessary in the preparation. For example, alkali metal salts such as sodium salt and potassium salt, alkaline earth metal salts such as calcium salt and magnesium salt, amine salts such as tri (n-butyl) amine salt, triethylamine salt, pyridine salt and amino acid salt Can do.

  The drug of the present invention can be used without particular limitation, such as hyaluronan having a certain molecular weight alone or a combination of hyaluronan having various molecular weights. The drug of the present invention comprises hyaluronan as an active ingredient, and improves endocrine disease and / or metabolic disease without adversely affecting the living body by administering an effective amount thereof to mammals including humans. Can do. Examples of endocrine diseases and metabolic diseases include diabetes. However, endocrine diseases are not limited to diabetes, but congenital goiter, hypothyroidism, hyperthyroidism, pituitary short stature, congenital adrenal hyperplasia, 21-hydroxylase deficiency, 11β -Hydroxylase deficiency, 3β-hydroxysteroid dehydrogenase deficiency, cholesterol-desmolase deficiency, 17α-hydroxylase deficiency, corticosterone-18-methyloxidase type 2 deficiency, male hypogonadism and secondary Examples include hypogonadism. The metabolic diseases are not limited to diabetes, but include galactosemia, congenital amino acid metabolism disorders, phenylketonuria, hyperphenylalaninemia, homocystinuria, maple syrup syndrome, maple syrup urine disease, cystic Fibrosis, nucleic acid metabolism disorder, lipid metabolism disorder, mucopolysaccharide metabolism disorder, Hurler syndrome, Hunter syndrome, Morquio syndrome, galactokinase deficiency UDP galactose 4-epimerase deficiency, fructose metabolism disorder, fructose-1,6-bisphosphatase Deficiency, pentoseuria, pyruvate metabolic disease and the like. That is, a drug containing hyaluronan as an active ingredient has a therapeutic effect on the various endocrine diseases and / or metabolic diseases described above.

  The agent of the present invention is administered hyaluronan or a salt thereof, as it is or as necessary, with a carrier, excipient, or other additive, orally or parenterally (intra-articular administration, intravenous, intramuscular, subcutaneous). As a pharmaceutical for administration (injection), enteral administration, transdermal administration, etc.), it can be formulated into any dosage form and administered to a patient by any administration method.

  Examples of oral preparations include solid preparations such as powders, granules, capsules and tablets; liquid preparations such as syrups, elixirs and emulsions. The powder can be obtained by mixing with excipients such as lactose, starch, crystalline cellulose, calcium lactate, calcium hydrogen phosphate, magnesium aluminate metasilicate, and silicic anhydride. In addition to the above excipients, the granule is optionally combined with a binder such as sucrose, hydroxypropylcellulose, polyvinylpyrrolidone, a binder such as carboxymethylcellulose, carboxymethylcellulose calcium, carboxymethylrose, carboxymethylcellulose calcium, etc. Further, a disintegrant may be added and granulated by wet or dry method. Tablets can be obtained by compressing the powder or granules as described above or by adding a lubricant such as magnesium stearate or talc. The tablet or granule is coated with an enteric base such as hydroxypropyl methylcellulose phthalate or methyl methacrylate copolymer, or coated with ethyl cellulose, carnauba wax, hardened oil, etc. can do. The hard capsule can be obtained by filling the above powder or granule into a hard capsule. Soft capsules can be obtained by mixing hyaluronan or a salt thereof with glycerin, polyethylene glycol, sesame oil, olive oil or the like and coating it with a gelatin film. A syrup can be obtained by dissolving a sweetener such as sucrose, sorbitol, and glycerin and hyaluronan or a salt thereof in water. In addition to sweeteners and water, essential oils, ethanol and the like can be added to make elixirs, or gum arabic, tragacanth, polysorbate 80, sodium carboxymethylcellulose and the like can be added to make emulsions or suspensions. Moreover, a corrigent, a coloring agent, a preservative, etc. can be added to these liquid preparations as needed.

  Examples of parenteral preparations include injections, rectal administration agents, pessaries, external preparations for skin, inhalants, aerosols, eye drops and the like. Injections include hyaluronan or salts thereof, pH regulators such as hydrochloric acid, sodium hydroxide, lactic acid, sodium lactate, sodium monohydrogen phosphate, sodium dihydrogen phosphate; isotonic agents such as sodium chloride and glucose; and It can be obtained by adding distilled water for injection, sterilizing and filtering and then filling ampoules. Further, mannitol, dextrin, cyclodextrin, gelatin and the like can be added and lyophilized in a vacuum to obtain an injection that can be dissolved at the time of use. Moreover, after adding an emulsifier, such as lecithin, polysorbate 80, polyoxyethylene hydrogenated castor oil, to hyaluronan or its salt, it can also be set as the emulsion for injection emulsified in water.

  For rectal administration, add a suppository base such as mono-, di- or triglyceride of cacao fatty acid or polyethylene glycol to hyaluronan or a salt thereof, dissolve by heating, and pour it into a mold to cool, Alternatively, hyaluronan or a salt thereof can be mixed with polyethylene glycol, soybean oil or the like and then coated with a gelatin film. The external preparation for skin can be obtained by adding white petrolatum, beeswax, liquid paraffin, polyethylene glycol or the like to hyaluronan or a salt thereof, and heating and kneading as necessary. The tape agent can be obtained by kneading hyaluronan or a salt thereof with a pressure-sensitive adhesive such as rosin or an alkyl acrylate polymer and spreading it on a nonwoven fabric or the like. The inhalant can be obtained by, for example, dissolving or dispersing hyaluronan or a salt thereof in a propellant such as a pharmaceutically acceptable inert gas and filling the pressure-resistant container.

  (Administration method) The administration method of the drug comprising the hyaluronan of the present invention as an active ingredient is not particularly limited, and examples thereof include intramuscular administration, intravenous administration and oral administration.

  (Dosage) The dosage is appropriately determined according to the target disease, the patient's age, health condition, body weight, etc. Generally, 0.05 to 50 mg / kg is divided once or more once a day. To administer.

  (Toxicity) The hyaluronan used in the present invention showed little or no cytotoxicity at doses showing biological activity as a drug.

  Hereinafter, although the chemical | medical agent which concerns on this invention is demonstrated in detail using an Example, the technical scope of this invention is not limited to a following example.

[Example 1]
In this example, HA4 was administered to KK-Ay mice, which are diabetes models, and the effects were examined.

Diabetes model animal 7-week-old male KK-Ay mice were purchased from Charles River Japan, and after 1 week of breeding, blood glucose level, urine glucose level, average body weight and SD were 8 The mice were grouped so that they did not differ. The group composition was a negative control (saline) group (intramuscular administration), HA4 (10 mg / kg) intramuscular administration group, and HA4 (10 mg / kg) intraperitoneal administration group.

Administration of test substance In this example, 2 mg / ml and 10 mg / ml HA4 were prepared. Specifically, HA4 is produced by the method of Tawada et al. (Tawada A, Masa T, Oonuki Y, Watanabe A, Matsuzaki Y, Asari A. Large-scale preparation, purification, and characterization of hyaluronan oligosaccharides from 4-mers to 52-mers. Glycobiology. 2002; 12 (7): 421-6.) And the concentration was adjusted with physiological saline. Moreover, physiological saline (saline) was adjusted as a control.

  In this example, intramuscular or intraperitoneal administration was performed once a day from the next day after diabetic model mice were grouped.

Blood glucose measurement and urine glucose measurement Blood glucose levels are collected from the tail at the time of grouping, 1 week and 2 weeks after administration, using a blood glucose meter (Exatech 2A / Medisense Region Plus <Medisense>) It was measured. The urine sugar value was measured using a urine test paper (Uriese, Terumo) at the time of grouping, 1 week and 2 weeks after administration. The blood glucose level and urine sugar level were statistically processed by Student's t-test or Tukey multiple comparison test.

Result 1) Blood glucose level (Figure 1)
One week and two weeks after administration, a significant increase in blood glucose level was observed in the negative control group. In the HA4-administered group, the increase in blood glucose level was slow in both the intramuscular administration group and the intraperitoneal administration group, and was not statistically significant compared to the grouping. From the results of FIG. 1, it was demonstrated that a drug containing hyaluronan as an active ingredient suppresses an increase in blood glucose level.

2) Urine sugar level (Figure 2)
One week and two weeks after administration, a significant increase in urinary glucose level was observed in the negative control group. In the HA4-administered group, the increase in urinary glucose level was slow in both the intramuscular administration group and the intraperitoneal administration group, and was not statistically significant compared to the grouping. Further, one week after administration, the urinary glucose level in the HA4 group was significantly lower than that in the negative control group. From the results of FIG. 1, it was demonstrated that a drug containing hyaluronan as an active ingredient suppresses an increase in urine sugar level and reduces the urine sugar value.

It is a graph which shows the result of having measured the blood glucose level (blood glucose level) when HA4 or a physiological saline is administered with respect to a diabetes model mouse. It is a graph which shows the result of having measured the urinary glucose level (urine sugar value) when HA4 or a physiological saline is administered with respect to a diabetes model mouse.

Claims (4)

  A therapeutic agent for endocrine diseases comprising hyaluronan as an active ingredient.   The hyaluronan is a tetrasaccharide containing two units of -D-glucuronic acid-β-1,3-DN-acetylglucosamine-β-1,4-. The therapeutic agent for endocrine disease according to 1.   A therapeutic agent for metabolic diseases comprising hyaluronan as an active ingredient.   The hyaluronan is a tetrasaccharide containing two units of -D-glucuronic acid-β-1,3-DN-acetylglucosamine-β-1,4-. 3. The metabolic disease therapeutic agent according to 3.

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