JPH0629184B2 - Osteoporosis treatment - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION [Field of Industrial Application] The object of the present invention is to provide a compound represented by the general formula (I): 2-phenyl-4H-1-benzopyran-4-one derivative represented by (wherein R ¹ and R ² are a hydrogen atom or an alkyl group having 1 to 3 carbon atoms) or a pharmacologically acceptable thereof The present invention provides a therapeutic agent for osteoporosis containing a salt capable of being treated.

[Conventional technology]

Osteoporosis refers to a pathological condition in which bone mass is reduced without affecting the chemical composition of bone, and its physiological characteristic is a decrease in bone proteins and calcium and phosphorus.

Osteoporosis increases with age and usually affects the spinal cord, causing low back pain and shortened height. In particularly advanced cases, long bones are also affected, and sometimes fractures occur.
It is said that most of the causes of femoral neck fractures in the elderly are due to senile osteoporosis.

There are various causes of this osteoporosis such as endocrine and nutritional disorders, and vitamin D preparations, calcium preparations, calcitonin preparations, phosphorus preparations, etc. are used as therapeutic agents, but their effects are not certain, so more effective There is a strong demand for the development of various formulations.

In recent years, a certain type of 3-
It has been reported that a phenyl-4H-1-benzopyran-4-one derivative (isoflavone derivative) has a bone resorption inhibitory effect and is useful as a therapeutic agent for osteoporosis (Japanese Patent Publication No. 54-1).
3391, JP 60-48924, JP 60-54379, JP
60-132917, JP-A-60-132976).

However, the 2-phenyl-4H-1-benzopyrans of the present invention
It has not been reported at all until now that a 4-one derivative (flavone derivative) exhibits a bone resorption inhibitory action and is useful as a therapeutic agent for osteoporosis.

[Problems to be Solved by the Invention] The 3-phenyl-4H-1-benzopyran-4-one derivative (isoflavone derivative) disclosed in the above-mentioned patent application has a weak bone resorption inhibitory action, and as a therapeutic agent for osteoporosis, I'm never satisfied. Therefore, we have found that benzopyran
As a result of diligent studies on the bone resorption inhibitory effect of the 4--4-one derivative, it was found that certain 2-phenyl-4H-1-benzopyran-4-
It has been found that an on derivative (flavone derivative) or a pharmacologically acceptable salt thereof has a strong bone resorption inhibitory action and an osteogenesis promoting action, and can be a more excellent therapeutic agent for osteoporosis.

The present invention is based on these findings.

[Means for solving problems]

The 2-phenyl-4H-1- represented by the general formula (I) of the present invention
The benzopyran-4-one derivative (flavone derivative) or a pharmacologically acceptable salt thereof exhibits a strong bone resorption inhibitory action and an osteogenesis promoting action, is highly safe, and is useful as a superior therapeutic agent for osteoporosis.

The 2-phenyl-4H-1- represented by the general formula (I) of the present invention
The benzopyran derivative is a known compound and can be produced according to the method described in the literature or a method similar thereto. For example, Organic Synthesis Collective Volume IV, pages 478-481 (Org.Syn.Coll.Vol.IV, 478
~ 481); Journal of the Chemical Society (JCS) 1926, 267 pages, Journal of the
It can be easily produced by the method of The Chemical Society (JCS), page 1936, 267 and Current Science, Volume 5, page 476, or the like, or a method similar thereto.

That is, 2-phenyl-4H- represented by the general formula (I)
The 1-benzopyran-4-one derivative has the formula (II) The acetophenone derivative represented by the formula (wherein R ¹ and R ² have the same meanings as described above) is reacted with benzoyl chloride to give a compound of formula (III) (In the formula, R ³ and R ⁴ are a benzoyl group or a carbon number of 1 to
After obtaining a benzooxyacetophenone derivative represented by (the alkyl group of 3), the formula (VI) is obtained by Baker VenKataraman rearrangement. (In the formula, R ³ and R ⁴ have the same meanings as described above), and a dibenzoylmethane derivative is prepared, and then dehydration ring closure is performed in acetic acid in the presence of sodium acetate.

In the present production method, when an acetophenone derivative in which R ¹ is a hydrogen atom is used as a raw material, a tribenzoylmethane derivative is produced in the Baker Bencatalaman rearrangement, so an acetophenone derivative in which R ^{1 is} an alkyl group, that is, 2-hydroxy It is preferable to use a -6-alkylacetophenone derivative as a raw material, and this alkoxy group can be easily converted into a hydroxy group by a conventional method after ring closure.

In the present production method, the acetophenone derivative represented by the general formula (II) used is a known compound and can be produced according to the method described in the literature.

The 2-phenyl-4H-1- represented by the general formula (I) of the present invention
A compound of a benzopyran-4-one derivative in which at least one of R ¹ and R ² is a hydrogen atom can be converted into a salt according to a conventional method. For example, the 2-phenyl-4H-1-benzopyran-4-one derivative represented by the general formula (I) of the present invention is added to an alcohol solution in which an equivalent amount of sodium hydroxide is dissolved, and after heating, The sodium salt can be obtained by concentrating under reduced pressure.

The 2-phenyl-4H-1- represented by the general formula (I) of the present invention
The benzopyran-4-one derivative can be made into a pharmaceutical preparation according to a conventional method. That is, pharmaceutical additives such as commonly used excipients, disintegrants, binders, lubricants and the like are mixed and prepared according to a conventional method to prepare various preparations such as tablets, powders and capsules. .

The 2-phenyl-4H-1- represented by the general formula (I) of the present invention
When a benzopyran-4-one derivative is used as a therapeutic agent for osteoporosis, about 10 to 1000 mg per day for an adult is made into an appropriate dosage form, such as tablets, powders, capsules, etc., and is orally administered in a divided dose or about 1 per adult per day. Approximately 100 mg is parenterally administered as an injection.

The 2-phenyl-4H-1- represented by the general formula (I) of the present invention
The benzopyran-4-one derivative showed strong bone resorption inhibitory action and osteogenesis promoting action in the in vitro experiment using chicken embryo femur, and the calcium and phosphorus contents in the bones of rats produced when a calcium-deficient diet was fed. It has the effect of remarkably improving the decrease in the content, and is useful as a therapeutic agent for osteoporosis.

〔Example〕

The following examples are given to further illustrate the present invention.
The melting points of the compounds in the examples are uncorrected.

Example 1 5,7-Dimethoxy-2-phenyl-4H-1-benzopyran-4-one (a) 2-benzoyloxy-4,6-dimethoxyacetophenone 2-hydroxy-4,6-dimethoxyacetophenone 2.5 g
Is dissolved in 12 m of dry pyridine, and 2.0 m of benzoyl chloride is added thereto while stirring under ice water. After stirring under cooling for 10 minutes, heat on a water bath for 20 minutes. After cooling, the reaction solution is added to ice-cooling and made weakly acidic with diluted hydrochloric acid. The result of precipitation is collected by filtration and recrystallized from methanol-water to obtain 3.3 g of 2-benzoyloxy-4,6-dimethoxyacetophenone.

Melting point: 76-80 ° C (b) 4,6-dimethoxy-2-hydroxybenzoylmethane 2-benzoyloxy-4,6-dimethoxyacetophenone 3.3
g is dissolved in 30 ml of dry N, N-dimethylformamide, and 60% sodium hydride (oil) is added with stirring under ice cooling. The mixture is stirred at room temperature for 3 hours. 200 reaction solution
m ice water and extract with benzene. The benzene layer is dried over anhydrous magnesium sulfate and concentrated under reduced pressure to obtain oily crude 4,6-dimethoxy-2-hydroxydibenzoylmethane.

(c) 5,7-Dimethoxy-2-phenyl-4H-1-benzopyran-4-
ON Crude 4,6-dimethoxy-2-hydroxybenzoylmethane obtained in (b) above, sodium acetate 15 g and acetic acid 60 m
The mixture is heated to reflux for 4 hours. 600m after reaction
The reaction solution is poured into ice water and the precipitated crystals are collected by filtration. After drying, dissolve in ether and dry over magnesium sulfate,
Concentrate to about half volume. The precipitated crystals are collected by filtration to obtain 800 mg of 5,7-dimethoxy-2-phenyl-4H-1-benzopyran-4-one.

Mp: 149-151 ° C. Elemental analysis: (as C ₁₇ H ₁₄ O ₄₎ Example 2 5,7-Dihydroxy-2-phenyl-4H-1-benzopyran-
4-one (a) 2,4,6-tribenzoyloxyacetophenone 2,4,6-
2.1 g of trihydroxyacetophenone was dissolved in 20 m of dry pyridine, and 6 m of benzoyl chloride was cooled in this solution under cooling.
And then heat for 30 minutes on a water bath. Ice water is added to the reaction solution, and diluted hydrochloric acid is added to make it weakly acidic. The acidic mixture is extracted with chloroform, dried over anhydrous magnesium sulfate and concentrated under reduced pressure. Methanol was added to the residue to dissolve it, and the precipitated crystals were collected by filtration under cooling to obtain 4.7 g of 2,4,6-tribenzoyloxyacetophenone.

Melting point: 114-116 ° C (b) 2,4-dibenzoyloxy-6-hydroxydibenzoylmethane 2,4,6-tribenzoyloxyacetophenone 4.7 g was dissolved in 30 m of dry N, N-dimethylformamide, To this is added 0.48 g of 60% sodium hydride (oil-based) while stirring with ice water. The mixture is stirred at room temperature for 3 hours. 300 reaction solution
m ice water, and then diluted hydrochloric acid to weakly acidify. . The precipitated crystals are collected by filtration to obtain 4.0 g of yellow crystals of 2,4-dibenzoyloxy-6-hydroxydibenzoylmethane.

(c) 5,7-Dihydroxy-2-phenyl-4H-1-benzopyran-4-one 2,4-dibenzoyloxy-6-hydroxydibenzoylmethane 4.7 g, sodium acetate 15 g and acetic acid 60 m
The mixture is heated to reflux for 4 hours. After completion of the reaction, the reaction solution is cooled, poured into 600 m of ice water, and the precipitated crystals are collected by filtration. The crystals obtained are washed with water and dissolved in 50 m of methanol. To this solution are added 5 g of potassium hydroxide and 10 m of water, and the mixture is heated on a boiling water bath for 1 hour. After completion of the reaction, the reaction solution is concentrated under reduced pressure, the residue is dissolved in 1.5 of water, and carbon dioxide gas is blown into this solution. The precipitated crystals were collected by filtration, dried and recrystallized from acetic acid-water to give 5,7-dihydroxy-2-phenyl-4H-1-.
0.1 g of benzopyran-4-one is obtained.

Melting point: 280-285 ° C Elemental analysis value: (as C ₁₅ H ₁₀ O ₄ ) Example 3 Bone resorption inhibitory effect Bone resorption inhibitory effect "composition culture application research method" page 111-1
14 (Osamu Yamane, Hiroyoshi Endo, Soft Science Publishing)
It measured according to the described method.

After removing femur bones of chicken embryos of 10 to 11 incubation and removing the soft tissues attached to the bone well, the phenol red containing the 2-phenyl-4H-1-benzopyran-4-one derivative of the present invention is not added. Pre-culture is carried out using 1 m of BGJb-HW2 culture solution (hereinafter referred to as culture solution) at 37 ° C. for 1 day by a rotary culture method. In addition, the compound of the present invention is prepared by directly diluting a solution of dimethyl sulfoxide dissolved in a culture medium by 1000 times to a concentration of 10 ^-4 molar.

The next day, ⁴⁵ CaCl ₂ was dissolved in a fresh culture solution to a concentration of 1 μCi / m, and the pre-cultured chicken embryo femur was immersed in 1 m of the solution.
Incubate with shaking at 37 ° C for 2 hours. As a result, bone mineral in the cultured bone is labeled with ⁴⁵ Ca. Immediately after completion of the culture
The cultured bone is washed with phosphate buffered saline that has been heated to 37 ° C to remove ⁴⁵ Ca attached to the bone.

The ⁴⁵ Ca-labeled cultured bone is again cultured in the culture medium by the rotary culture method (10 revolutions / hour). Accurately collect a fixed amount of the culture solution every 12, 24, 48, 72 hours, discard the remaining culture solution and add fresh culture solution at the same time. The separated ⁴⁵ Ca radioactivity in the culture broth was measured with a liquid scintillation counter, it converts the radioactivity ⁴⁵ Ca in the whole broth.
After completion of the culture, the bone tissue is allowed to stand in 1N hydrochloric acid for 1 day to elute total calcium, and its radioactivity is measured to obtain the final residual radioactivity in the cultured bone.

From the obtained measurement value, calculate the ratio of the radioactivity remaining in the cultured bone to the total radioactivity that was initially taken into the bone tissue, and calculate the radioactivity residual rate decay curve in the cultured bone after 24 hours. The elution of bone mineral by osteoclasts is directly returned, and the turnover rate of calcium in bone mineral deposited on the cultured bone is determined as the biological half-life T1 / 2 from the obtained linear gradient.

The compound group of the present invention and the control group each consisted of 5 cases.

T1 of the compound group of the present invention compared to the value of T1 / 2 of the control group
A large value of / 2 indicates that the compound of the present invention has a bone resorption inhibitory effect, and the efficacy of the compound of the present invention for a bone resorption inhibitory effect was determined by the following formula.

The results are shown below.

Example 4 Bone formation promoting action The bone formation promoting action is shown in "Tissue culture applied research method" pages 103-1.
11 (Osamu Yamane, edited by Hiroyoshi Endo, published by Soft Science,
(1985).

The femur of a chicken embryo on the 9th day of incubation was extracted, and the soft tissue attached to the bone was removed well. group,
Using the other as a control group, attach each to the inner surface of a rectangular test tube for culture, add 2 m of BGJb-HW2 culture solution (hereinafter referred to as culture solution) to it, seal with a silicon stopper, and spin culture at 37 ° C (10 Rotate / time). The compound of the present invention is dissolved in dimethyl sulfoxide and then directly diluted 1000-fold in a culture solution to a concentration of 10 ^-4 molar. Bone culture is continued for 6 days while measuring the bone length every day and replacing with fresh culture medium.

This experiment was carried out in 6 cases in each group.

At the end of culturing, the cultured bone was washed with phosphate buffered saline, 1
Calcium is eluted from the bone tissue by allowing it to stand in normal hydrochloric acid for 1 day, and the amount of eluted Ca is determined by the chelate method using orthocresolphthalein.

The potency of the osteogenesis promoting action of the compound of the present invention was determined by the following formula.

The results are shown below.

Example 5 Effect on Calcium / Phosphorus Content in Bone of Rats Fed with Ca-Deficient Diet Twenty 3-week-old male Wistar rats were divided into two groups, 10 rats per group, and a Ca-deficient diet was added to each group. The other group was fed with a Ca-deficient diet and 5,7-dimethoxy-2-phenyl-4H-1-benzopyran-4-one 300 mg / kg of the present invention and bred for 2 weeks to obtain calcium and phosphorus in the femur. The quantity was measured.

The results are shown below.

Example 6 Acute toxicity 5,7-dihydroxy-2-phenyl-4H-1-benzopyran-
4-on was applied to CMC, and 7 weeks old ICR mice 10 each
Using a rat, 1000, 2000, 3000 mg / kg was orally administered and observed for 7 days. There were no deaths in any group and no toxic symptoms were observed.

Example 7 Preparation of preparation (a) Tablet 5,7-dihydroxy-2-phenyl-4H-1-benzopyran-
4-Gone 100 g, lactose 95 g, corn starch 40 g are mixed, and then 5% hydroxypropyl cellulose aqueous solution is added and kneaded, followed by drying. Mold into 1000 tablets.

(b) Capsule 5,7-dihydroxy-2-phenyl-4H-1-benzopyran-
4-on 100g, lactose 39g and corn starch 35
6 g of talc is added to the mixture and mixed, and then 1000 hard capsules are filled.

〔The invention's effect〕

The 2-phenyl-4H-1-benzopyran-4H-1-one derivative represented by the general formula (I) of the present invention and the pharmacologically acceptable salts thereof are used in in vitro experiments using chicken embryo femurs. , Shows the effect of suppressing bone resorption and promoting bone formation, and
It markedly suppresses the decrease in the calcium and phosphorus contents in the bones of the rat, which occurs when a calcium-deficient diet is fed.

Therefore, 2-phenyl-4 represented by the general formula (I) of the present invention
The H-1-benzopyran-4-one derivative is useful as a therapeutic agent for osteoporosis.

Claims (3)

[Claims] 1. A general formula 2-phenyl-4H-1-benzopyran-4-one derivative represented by (wherein R ¹ and R ² are a hydrogen atom or an alkyl group having 1 to 3 carbon atoms) or a pharmacologically acceptable thereof A therapeutic agent for osteoporosis, which comprises the resulting salt as an active ingredient. 2. A formula The therapeutic agent for osteoporosis according to claim 1, which comprises a 2-phenyl-4H-1-benzopyran-4-one derivative represented by the following as an active ingredient. 3. A formula The therapeutic agent for osteoporosis according to claim 1, which comprises a 2-phenyl-4H-1-benzopyran-4-one derivative represented by or a pharmacologically acceptable salt thereof as an active ingredient.