JP2005263742A - Integrin appearance enhancer - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a medicine for enhancing the intrauterine appearance of integrin α5β1. <P>SOLUTION: There are provided an integrin α5β1 intrauterine appearance enhancer containing danazol as the effective component and a therapeutic agent containing danazol as the effective component and for curing infertility caused by the decrease in integrin α5β1 intrauterine appearance. <P>COPYRIGHT: (C)2005,JPO&NCIPI

Description

  The present invention relates to a pharmaceutical that enhances in utero expression of integrin α5β1, which is a cell adhesion molecule.

  Integrins have been identified as molecules that adhere to the extracellular matrix. Subsequent studies have shown that integrins not only adhere to the extracellular matrix on the cell surface, but also interact with many intracellular skeletal molecules in the cell, and are also involved in signal transduction into the cell. It was found that it plays an important role as a regulator of cell function.

  Integrins are involved in cell body development and differentiation, tissue support and construction, migration of cells such as leukocytes into blood vessels, represented by migration into the tissue, cell recognition and information transmission in the tissue, etc. It plays a central role in all the basic functions in the body, and also plays an important role in inflammation, blood coagulation, tumor metastasis, etc. caused by autoimmunity and bacterial infection (Non-patent literature) 1).

  Regarding integrin, its application to the treatment of leukocyte adhesion inhibition, platelet aggregation inhibition, cancer metastasis inhibition, myocardial infarction, arteriosclerosis, and osteolytic disease by suppressing its expression is being investigated. In addition, it is expected to be effective in suppressing cancer metastasis and treating arteriosclerosis by enhancing its expression (Patent Document 1).

It has also been reported that a physiologically active peptide having an integrin activating action is used for tissue repair such as wound healing and surgery, improvement of obesity, etc. (Patent Document 2).
Integrins are transmembrane proteins that form heterodimers composed of α and β chains. About 20 types of integrins have been reported so far by combining 15 types of α chains and 8 types of β chains. . The extracellular matrix to be bound differs depending on each combination. For example, it is known that integrin α5β1 binds to fibronectin, integrin α6β1 binds to laminin, and integrin αvβ3 binds to vitronectin. On the other hand, integrin α2β1 binds to a plurality of extracellular substrates such as type 1 and type 4 collagen, laminin and fibronectin. Of these, integrin α5β1 is known to be deeply involved in the implantation of fertilized eggs (Non-patent Document 2).

Danazol exhibits an anti-gonadotropin-releasing factor action (Non-patent Document 3) and is used for the treatment of endometriosis and mastopathy. It is known to administer danazol to infertile patients and danazol as a pretreatment for in vitro fertilization. In this case, danazol is used to treat endometriosis, which often occurs in infertile patients. To be administered. However, the specific relationship between danazol and integrin is not known, and it is not known that danazol improves the implantation rate of fertilized eggs and increases the success rate of pregnancy.
Tanaka Yoshiya, Clinical Immunity, Vol. 29, No. 7, pp. 823-831, 1997 JP-A-11-246428 JP 2001-213898 A Masayuki Ozawa, Clinical Immunity, Vol. 25, No. 9, 1159-1164, 1993 THE MERCK INDEX THIRTEENTH EDITION, 2838, P. 493, 2001

  An object of the present invention is to provide a pharmaceutical that can be used for treatment of infertility by enhancing in-uterine expression of integrin α5β1 or a reagent for analyzing the function of integrin α5β1.

  As a result of diligent efforts to solve the above-mentioned problems, the present inventors have found that danazol enhances the expression of integrin α5β1 in the uterus, thereby completing the present invention.

  That is, the gist of the present invention resides in an integrin α5β1 in-utero expression enhancer, an infertility therapeutic agent, and a pregnancy rate improver characterized by containing danazol as an active ingredient. A preferred embodiment of the pregnancy rate improving agent in the present invention is a pregnancy rate improving agent in natural pregnancy that does not undergo a series of artificial operations such as artificial insemination, in vitro fertilization and embryo transfer.

  Danazol has the effect of enhancing the in-uterine expression of integrin α5β1, the effect of treating infertility due to decreased integrin α5β1 in-utero expression, and the effect of improving the pregnancy rate.

  The administration form in the case of using the integrin α5β1 intrauterine expression enhancer according to the present invention for infertility treatment is arbitrary. For example, a preparation in a form well known to those skilled in the art prepared by adding pharmacologically and pharmaceutically acceptable additives to danazol, orally administered by tablets, capsules, etc., or injections, suppositories, etc. By parenteral administration.

  Examples of pharmacologically and pharmaceutically acceptable additives include excipients such as glucose, lactose, D-mannitol, starch or crystalline cellulose; disintegrants or disintegration such as carboxymethylcellulose, starch or carboxymethylcellulose calcium Adjuncts; binders such as hydroxypropylcellulose, hydroxypropylmethylcellulose, polyvinylpyrrolidone or gelatin; lubricants such as magnesium stearate or talc; coating agents such as hydroxypropylmethylcellulose, sucrose, polyethylene glycol or titanium oxide; petrolatum, fluid Bases such as paraffin, polyethylene glycol, gelatin, kaolin, glycerin, purified water or hard fat; aqueous or used such as distilled water for injection, physiological saline or propylene glycol Dissolving agent or solubilizing agent capable of preparing dissolution type injections; isotonic agents such as glucose, sodium chloride, D-mannitol, or glycerin; pH adjustment of inorganic acids, organic acids, inorganic bases, organic bases, etc. Agents; pigments; diluents; stabilizers; propellants and pressure-sensitive adhesives.

  What is necessary is just to select a dosage suitably according to a patient. For example, 10 to 1000 mg, preferably 50 to 600 mg, is administered per day, and 1 to 100 mg, preferably 5 to 60 mg, is administered by injection.

In addition, since ovulation may be suppressed during danazol administration, in order to improve the pregnancy rate of spontaneous pregnancy, it is necessary to suspend danazol administration for a period corresponding to at least one sexual cycle. In such a case, a cycle consisting of a period of continuous administration of danazol for 1 to 180 days, preferably 30 to 120 days, and a drug holiday of 30 to 180 days, preferably 90 to 180 days, may be repeated.

  Hereinafter, the present invention will be described more specifically with reference to examples. However, the scope of the present invention is not limited to these examples.

Danazol is a 7-week-old female Balb / c mouse (Japan SLC Co., Ltd.) acclimated to the environment for 1 week under normal conditions, and then dissolved in cottonseed oil containing 10% ethanol subcutaneously on the back of the mouse. Administration was once a day for 2 weeks. Two weeks is a period of continuous administration over three sexual cycles because the sexual cycle of mice is approximately 4 days (early estrus, estrus, late estrus, estrus resting period).
(Example 1)
One group of 10 Balb / c mice received 0 or 7.5 mg / kg (BW) of danazol, respectively. After completion of the administration, the animals were allowed to live with the male in the evening after a rest period of 4 days corresponding to one sexual cycle, and the presence or absence of mating was observed by looking at the vaginal plug the next morning. The day when the vaginal plug was observed was defined as day 0 of pregnancy, and Evans Blue 10 mg / mL was injected from the tail vein on the 5th day of pregnancy in the implantation period. The site was dissected 30 minutes later, and the site where the uterus was thickened and colored with Evans blue was used as the implantation site. The number of implantation sites was almost the same in the danazol administration group and the control group (no danazol administration). The pregnancy success rate calculated from the presence or absence of the implantation site was 84.6% in the danazol administration group and 46.2% in the control group. This indicates that danazol has the effect of improving spontaneous pregnancy in mice.
(Example 2)
One group of 15 Balb / c mice received 0 or 7.5 mg / kg (BW) of danazol, respectively. After administration of danazol, a section was prepared from a uterine fixed with formalin / paraffin embedded according to a conventional method, and the localization of integrin α5-positive cells in the uterus was examined by an immunohistochemical method. As the primary antibody, integrin α5 antibody (C-19, Santa Cruz, Biotech) was used. Vectastain Elite ABC kit (Vector Lab.) Was used for the process after the secondary antibody. DAB kit (Vector Lab.) Was used for color development. The results of immunohistochemistry are shown in FIG. FIG. 1 shows that danazol enhances in uterine expression of integrin α5β1.

  Danazol has the action of enhancing in utero expression of integrin α5β1, and therefore can be used alone or in combination with a compound having an integrin activation action for infertility treatment.

  Danazol can also be used for functional analysis of integrin α5β1.

It is a figure which shows the influence which danazol administration has on the in-utero expression of integrin (alpha) 5 (beta) 1.

Claims (4)

An integrin α5β1 uterine expression enhancer comprising danazol as an active ingredient. A therapeutic agent for infertility caused by decreased expression of integrin α5β1 in utero containing danazol as an active ingredient. A pregnancy rate improver containing danazol as an active ingredient. The pregnancy rate improving agent according to claim 3, wherein the pregnancy is natural pregnancy.