JP2007063193A - Inhibitor of hyaluronic acid synthesis - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an inhibitor of hyaluronic acid synthesis as a new medicinal application of a coumarin-based compound except MU (4-Methylumbelliferone). <P>SOLUTION: The inhibitor of the hyaluronic acid synthesis contains the coumarin-based compound represented by the general formula (wherein, R<SP>1</SP>and R<SP>2</SP>are the same or different, and each hydrogen or an alkyl group with the proviso that at least one thereof is an alkyl group; R<SP>3</SP>, R<SP>4</SP>, R<SP>5</SP>and R<SP>6</SP>are the same or different, and each hydrogen or a hydroxy group with the proviso that at least one thereof is a hydroxy group; with the proviso that the case wherein R<SP>1</SP>is hydrogen, R<SP>2</SP>is a methyl group, R<SP>3</SP>is hydrogen, R<SP>4</SP>is hydrogen, R<SP>5</SP>is a hydroxy group and R<SP>6</SP>is hydrogen is excluded) of the figure or a pharmacologically acceptable salt thereof as an active ingredient. <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

  The present invention relates to a hyaluronic acid synthesis inhibitor.

  4-Methylumbelliferone (MU), one of the coumarin compounds represented by the following chemical structural formula, has long been known for medicinal use as a bile agent, Have been reported to have a synthetic inhibitory effect on hyaluronic acid (Non-patent Document 1).

Hyaluronic acid is a linear polymer polysaccharide composed of a repeating structure of disaccharide units of glucuronic acid and N-acetylglucosamine, and is widely distributed in the living body as one of the main components of the extracellular matrix. In recent years, it has been known that this hyaluronic acid has a great influence not only as a tissue construction material but also on cell behavior. One example is the relationship between cancer cells and hyaluronic acid. Cancer tissue is known to form a hyaluronic acid-rich environment, and it has been suggested that there is a relationship between the proliferation and invasion of cancer cells and hyaluronic acid. In fact, it is known that in a culture system, when cancer cells and fibroblasts are mixed and cultured, hyaluronic acid synthesis in fibroblasts is remarkably increased. As described above, it is known that MU has an inhibitory effect on hyaluronic acid synthesis, but there has been no report on the action of coumarin compounds other than MU on the synthesis of hyaluronic acid.
Nakamura, T., Takagaki, K., Shibata, S., Tanaka, K., Higuchi, T., and Endo, M. (1995) Biochem. Biophys. Res. Commun., 208, 470-475

  Then, this invention aims at providing the hyaluronic acid synthesis inhibitor as a novel pharmaceutical use of coumarin type compounds other than MU.

  The hyaluronic acid synthesis inhibitor of the present invention, which has been obtained as a result of repeated studies by the present inventors in view of the above points, is, as described in claim 1, a coumarin compound represented by the following general formula or a pharmaceutical product thereof: It is characterized by using a salt that is acceptable as an active ingredient.

(Wherein R ¹ and R ² are the same or different and represent hydrogen or an alkyl group, but at least one is an alkyl group. R ³ , R ⁴ , R ⁵ and R ⁶ are the same or different and represent hydrogen or a hydroxyl group. Wherein at least one is a hydroxyl group, except that R ¹ is hydrogen, R ² is a methyl group, R ³ is hydrogen, R ⁴ is hydrogen, R ⁵ is a hydroxyl group, and R ⁶ is hydrogen)
The hyaluronic acid synthesis inhibitor according to claim 2 is the hyaluronic acid synthesis inhibitor according to claim 1, wherein at least two of the coumarin compounds R ³ , R ⁴ , R ⁵ and R ⁶ are hydroxyl groups. Features.
The hyaluronic acid synthesis inhibitor according to claim 3 is the hyaluronic acid synthesis inhibitor according to claim 2, wherein the coumarin compound is a 5,7-dihydroxy-4-alkylcoumarin or 6,7-dihydroxy-4-alkyl. It is a coumarin.
The hyaluronic acid synthesis inhibitor according to claim 4 is the hyaluronic acid synthesis inhibitor according to claim 3, wherein the coumarin compound is 5,7-dihydroxy-4-methylcoumarin or 6,7-dihydroxy-4-methyl. It is a coumarin.
Further, the hyaluronic acid synthesis inhibiting composition of the present invention is characterized in that, as described in claim 5, it contains a coumarin compound represented by the following general formula or a pharmaceutically acceptable salt thereof as an active ingredient. To do.

(Wherein R ¹ and R ² are the same or different and represent hydrogen or an alkyl group, but at least one is an alkyl group. R ³ , R ⁴ , R ⁵ and R ⁶ are the same or different and represent hydrogen or a hydroxyl group. Wherein at least one is a hydroxyl group, except that R ¹ is hydrogen, R ² is a methyl group, R ³ is hydrogen, R ⁴ is hydrogen, R ⁵ is a hydroxyl group, and R ⁶ is hydrogen)

  According to the present invention, a hyaluronic acid synthesis inhibitor is provided as a novel pharmaceutical use of coumarin compounds other than MU.

  The hyaluronic acid synthesis inhibitor of the present invention comprises a coumarin compound represented by the following general formula or a pharmaceutically acceptable salt thereof as an active ingredient. The hyaluronic acid synthesis inhibitor of the present invention has an effect of inhibiting the proliferation, invasion, and metastasis of cancer cells by inhibiting the synthesis of hyaluronic acid in cancer cells, and thus is useful for the prevention and treatment of cancer. Further, the hyaluronic acid synthesis inhibitor of the present invention is used for various fibrotic diseases caused by excessive production of hyaluronic acid, for example, chronic inflammatory diseases in various organs such as liver, lung, and kidney including chronic hepatitis. It is also useful for the prevention and treatment of accompanying fibrosis and pleural mesothelioma.

(Wherein R ¹ and R ² are the same or different and represent hydrogen or an alkyl group, but at least one is an alkyl group. R ³ , R ⁴ , R ⁵ and R ⁶ are the same or different and represent hydrogen or a hydroxyl group. Wherein at least one is a hydroxyl group, except that R ¹ is hydrogen, R ² is a methyl group, R ³ is hydrogen, R ⁴ is hydrogen, R ⁵ is a hydroxyl group, and R ⁶ is hydrogen)

  In the above general formula, the alkyl group is preferably linear or branched having 1 to 6 carbon atoms. Specifically, methyl group, ethyl group, n-propyl group, isopropyl group, n-butyl group, isobutyl group, sec-butyl group, tert-butyl group, n-pentyl group, isopentyl group, n-hexyl group, etc. Is exemplified.

  Examples of the pharmaceutically acceptable salt of the coumarin compound represented by the above general formula include alkali metal salts such as sodium salt and potassium salt.

Among the coumarin compounds represented by the above general formula, coumarin compounds in which at least two of R ³ , R ⁴ , R ⁵ and R ⁶ are hydroxyl groups are more effective in inhibiting hyaluronic acid synthesis and water-soluble than MU. Therefore, it is suitable as an active ingredient of the hyaluronic acid synthesis inhibitor of the present invention. Specific examples of such a compound include 5,7-dihydroxy-4-alkylcoumarin represented by 5,7-dihydroxy-4-methylcoumarin and 6,7-dihydroxy-4-methylcoumarin. Examples include 6,7-dihydroxy-4-alkylcoumarin. The coumarin compounds represented by the above general formulas are described in, for example, Kawase et al. , In vivo 17: 509-512 (2003), some of which are also commercially available.

  The hyaluronic acid synthesis inhibitor of the present invention can be administered orally or parenterally (for example, intravenous injection, subcutaneous administration, rectal administration, etc.). What is necessary is just to formulate in the dosage form suitable for each administration method in the case of administration. Examples of the dosage form include tablets, capsules, granules, powders, fine granules, pills, troches, sublingual tablets, suppositories, ointments, injections, emulsions, suspensions, syrups and the like. These preparations are non-toxic excipients, binders, lubricants, disintegrants, preservatives, isotonic agents, stabilizers, dispersants, antioxidants, colorants, flavoring agents, buffers It can carry out by a method known per se using an additive such as an agent. Non-toxic additives include, for example, starch, gelatin, glucose, lactose, fructose, maltose, magnesium carbonate, talc, magnesium stearate, methylcellulose, carboxymethylcellulose, gum arabic, polyethylene glycol, propylene glycol, petrolatum, glycerin, ethanol Syrup, sodium chloride, sodium sulfite, sodium phosphate, citric acid, polyvinylpyrrolidone, water and the like. In addition, in order to reinforce or enhance the usefulness of the present invention, other drugs may be contained in the preparation.

  The content of the active ingredient in the preparation varies depending on the dosage form, but generally a concentration of 0.1 to 100% by weight is desirable. The dosage of the preparation can be widely adjusted according to the gender, age and weight of the administration subject, the severity of symptoms, the diagnosis of a doctor, etc., but in general, it should be 0.01 to 300 mg / kg per day. it can. The above dose may be administered once or divided into several times a day.

  EXAMPLES Hereinafter, although an Example demonstrates this invention in detail, this invention is limited to the following description and is not interpreted at all.

Example 1: Evaluation of inhibitory effect on hyaluronic acid synthesis of human pancreatic cancer cells (experimental method)
1. Cultured cells Human pancreatic cancer cells KP-1NL cultured in RPMI-1640 containing 10% fetal calf serum (Biofluid, Australia) in 5% CO ₂ -95% air at 37 ° C. were used.

2. Evaluation sample The following compounds were used as evaluation samples.
Compound 1 (Comparative Example): Coumarin Compound 2 (Comparative Example): 7-hydroxycoumarin Compound 3 (Comparative Example): 7-methoxy-4-methylcoumarin Compound 4 (Comparative Example): 6,7-dihydroxy Coumarin Compound 5 (Example): 6-hydroxy-4-methylcoumarin Compound 6 (Example): 7-hydroxy-3-methylcoumarin Compound 7 (Example): 7-hydroxy-3,4-dimethyl Coumarin Compound 8 (Example): 5,7-dihydroxy-4-methylcoumarin Compound 9 (Example): 6,7-dihydroxy-4-methylcoumarin Compound 10 (Reference Example) 4-methylumbellite Ferron (MU)
Compounds 1, 2, and 10 were purchased from Wako Pure Chemical Industries. Compounds 3, 5, and 9 were purchased from Tokyo Chemical Industry. Compound 4 was purchased from Alfa Aesar. Compounds 6 and 7 were prepared according to Kawase et al. , In vivo 17: 509-512 (2003). Compound 8 was purchased from SSX.

3. Quantitative determination of hyaluronic acid in culture medium 2.0 × 10 ⁴ KP-1NL were plated on a 10 cm culture dish and incubated for 24 hours. After 24 hours, an evaluation sample dissolved in dimethyl sulfoxide in the medium was added so that its concentration was 1.0 × 10 ⁻² mM. After 72 hours, the culture medium and cells were collected, and hyaluronic acid per 10 ⁶ cells secreted into the culture medium was quantified using a hyaluronic acid measurement kit (Seikagaku Corporation).

(Experimental result)
The results are shown in Table 1 (the amount of hyaluronic acid in the medium is an average value of n = 6).

As is clear from Table 1, Compounds 5 to 9 effectively inhibited the hyaluronic acid synthesis of cancer cells as compared to the control to which no evaluation sample was added. Among them, compounds 8 and 9 show an inhibitory action superior to that of MU, and have higher water solubility than MU (according to a separate experiment), so that they are suitable as active ingredients of the hyaluronic acid synthesis inhibitor of the present invention. I found out. None of the evaluation samples affected the cell proliferation activity at a concentration of 1.0 × 10 ⁻² mM (by a separate experiment).

Example 2 Evaluation of Inhibitory Action on Human Hyaluronic Acid Synthesis of Human Colon Cancer Cells The same procedure as in Example 1 was conducted except that human colon cancer cell DLD-1 was used instead of human pancreatic cancer cell KP-1NL. The results are shown in Table 2 (the amount of hyaluronic acid in the medium is an average value of n = 6).

  As is clear from Table 2, even when the human colon cancer cell DLD-1 was used, the same results as when the human pancreatic cancer cell KP-1NL was used could be obtained.

Formulation Example 1: Tablet A 200 mg tablet having the following component composition containing 5 mg of 5,7-dihydroxy-4-methylcoumarin per tablet was prepared by mixing each component well and then tableting.
5,7-dihydroxy-4-methylcoumarin 5mg
Lactose 137〃
Starch 45〃
Carboxymethylcellulose 10〃
Talc 2〃
Magnesium stearate 1〃
200mg / tablet

Formulation Example 2: Capsule 100 mg capsules comprising the following component composition containing 20 mg of 6,7-dihydroxy-4-methylcoumarin per capsule were prepared by thoroughly mixing the components and filling the capsules. .
6,7-dihydroxy-4-methylcoumarin 20mg
Lactose 53〃
Starch 25〃
Magnesium stearate 2〃
Total 100mg / capsule

  The present invention has industrial applicability in that it can provide a hyaluronic acid synthesis inhibitor as a novel pharmaceutical use of a coumarin compound other than MU.

Claims (5)

A hyaluronic acid synthesis inhibitor comprising a coumarin compound represented by the following general formula or a pharmaceutically acceptable salt thereof as an active ingredient.

(Wherein R ¹ and R ² are the same or different and represent hydrogen or an alkyl group, but at least one is an alkyl group. R ³ , R ⁴ , R ⁵ and R ⁶ are the same or different and represent hydrogen or a hydroxyl group. Wherein at least one is a hydroxyl group, except that R ¹ is hydrogen, R ² is a methyl group, R ³ is hydrogen, R ⁴ is hydrogen, R ⁵ is a hydroxyl group, and R ⁶ is hydrogen) The hyaluronic acid synthesis inhibitor according to claim 1, wherein at least two of R ³ , R ⁴ , R ⁵ and R ^{6 in} the coumarin compound are hydroxyl groups.   The hyaluronic acid synthesis inhibitor according to claim 2, wherein the coumarin compound is 5,7-dihydroxy-4-alkylcoumarin or 6,7-dihydroxy-4-alkylcoumarin.   The hyaluronic acid synthesis inhibitor according to claim 3, wherein the coumarin compound is 5,7-dihydroxy-4-methylcoumarin or 6,7-dihydroxy-4-methylcoumarin. A hyaluronic acid synthesis inhibiting composition comprising a coumarin compound represented by the following general formula or a pharmaceutically acceptable salt thereof as an active ingredient.

(Wherein R ¹ and R ² are the same or different and represent hydrogen or an alkyl group, but at least one is an alkyl group. R ³ , R ⁴ , R ⁵ and R ⁶ are the same or different and represent hydrogen or a hydroxyl group. Wherein at least one is a hydroxyl group, except that R ¹ is hydrogen, R ² is a methyl group, R ³ is hydrogen, R ⁴ is hydrogen, R ⁵ is a hydroxyl group, and R ⁶ is hydrogen)