JPH0629185B2 - 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 or a pharmacologically acceptable salt thereof (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 disclosed in the above-mentioned patent application has a weak bone resorption inhibitory action and is not completely satisfactory as a therapeutic agent for osteoporosis. Therefore, the present inventors have conducted extensive studies on the bone resorption inhibitory effect of benzopyran-4-one derivatives, and found that certain 2-phenyl-4H-1-benzopyran-4-one derivatives or their pharmacologically It has been found that an acceptable salt has a strong bone resorption inhibitory action and a bone formation 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) exhibits a strong bone resorption inhibitory action and an osteogenesis promoting action, and is useful as a highly safe therapeutic agent for osteoporosis.

The 2-phenyl-4H-1- represented by the general formula (I) of the present invention
The benzopyran-4-one derivative is a known compound, and can be produced according to a method described in the literature or a method similar thereto. For example, Organic Synthesis Collective Volume IV, pages 478-481 (Org.Syn.Col.
l.IV, 478-481); Chemis Berhitte (Chim.Ber.) 43
Volume, page 1966; Kamical Abstract (Chem.Abst
r.) 26, 4816 (1932) or the like or a method similar thereto.

That is, 2-phenyl-4H-1-benzopyran-4-one represented by the general formula (I) of the present invention, 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 the 2-benzoyloxyacetophenone derivative represented by the formula (VI) is obtained by Baker Venlataramanrearrangement. A dibenzoylmethane derivative represented by the formula (wherein R ³ and R ⁴ have the same meanings as described above), and then heated in acetic acid in the presence of sodium acetate, and then hydrolyzed if necessary. be able to.

The 2-phenyl-4H-1- represented by the general formula (I) of the present invention
Benzopyran-4-one derivative in which R ¹ and R ² have 1 carbon atoms
The compound having an alkyl group of 3 to 3 is 2-phenyl-4H- in which at least one of R ¹ and R ² is a hydrogen atom.
A 1-benzopyran-4-one derivative having 1 to 1 carbon atoms according to a conventional method
It can also be produced by reacting with the alkyl halide of 3.

Further, the 2-phenyl-4H-1-benzopyran-4-one derivative represented by the general formula (I) of the present invention in which R ¹ and R ² are hydrogen atoms is at least R ¹ and R ² . It can also be produced by treating a 2-phenyl-4H-1-benzopyran-4-one derivative, one of which is an alkyl group having 1 to 3 carbon atoms, under acidic conditions.

In the present production method, the above-mentioned general formula (I
The acetophenone derivative represented by I) is a known compound and can be produced by the method described in the literature.

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

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

The 2-phenyl-4H-1- represented by the general formula (I) of the present invention
When the 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 day per adult. Administer ~ 100mg parenterally as an injection, etc.

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 6-Hydroxy-7-methoxy-2-phenyl-4H-1-benzopyran-4-one (a) 2,5-dibenzoyloxy-4-methoxyacetophenone 2,5-dihydrooxy-4- Methoxy acetophenone 5.0 g
Is dissolved in 10 m of dry pyridine, and 8.5 g of benzoyl chloride is gradually added to this solution with stirring. The mixture is heated on a boiling water bath for 10 minutes. After completion of the reaction, the reaction solution is cooled, diluted hydrochloric acid is added to make it weakly acidic, and the precipitated crystals are collected by filtration. The obtained crystals were washed with water and recrystallized from methanol / chloroform,
9.3 g of white crystals of 2,5-dibenzoyloxy-4-methoxyacetophenone are obtained.

Melting point: 233 to 234.5 ° C (b) 5-benzoyloxy-2-hydroxy-4-methoxydibenzoylmethane 2,5-dibenzoyloxy-4-methoxyacetophenone 20
g is dissolved in a mixture of 30 m dry N, N-dimethylformamide and dry pyridine. While heating the solution to 50 ° C., 4.0 g of sodium amide are added with stirring. The mixture is reacted under stirring at 100 ° C. for 1 hour. After completion of the reaction, the reaction solution is cooled, poured into water of 1 and made weakly acidic with dilute hydrochloric acid. The precipitated crystals were collected by filtration and recrystallized from methanol-chloroform to give 5-benzoyloxy-2-hydroxy-4-
4.0 g of methoxydibenzoylmethane are obtained.

Melting point: 180-182 ° C (c) 6-benzoyloxy-7-methoxy-2-phenyl-4H-1-
Benzopyran-4-one 5-benzooxy-2-hydroxy-4-methoxydibezoylmethane 4.0 g, anhydrous sodium acetate 4.0 g and acetic acid 25
The mixture of m is heated to reflux for 3 hours. After completion of the reaction, the reaction solution is poured into 100 m of ice water and the precipitated crystals are collected by filtration.
The obtained crystals were recrystallized from methanol-chloroform,
3.6 g of 6-benzoyloxy-7-methoxy-2-phenyl-4H-1-benzopyran-4-one are obtained.

Melting point: 233-235 ° C (d) 6-hydroxy-7-methoxy-2-phenyl-4H-1-benzopyran-4-one 6-benzoyloxy-7-methoxy-2-phenyl-4H-1-benzopyran- 3.3 g of 4-one is suspended in 200 m of methanol, and 5 m of an aqueous solution containing 3.5 g of potassium hydroxide is added to the suspension. The mixture is heated at reflux for 2 hours. After completion of the reaction, methanol is distilled off under reduced pressure. The residue is added to water, carbon dioxide gas is blown in, and the precipitated crystals are collected by filtration. The obtained crystals were recrystallized from ethanol-methanol to give 6-hydroxy-7-methoxy-2-phenyl-4H-1-benzopyran-4.
2.5 g of H-1-one are obtained.

Melting point: 208-210 ° C Elemental analysis value: (as C ₁₆ H ₁₂ O ₄ ) Example 2 6,7-Dimethoxy-2-phenyl-4H-1-benzopyran-4-one 6-Hydroxy-7-methoxy-2-phenyl-4H-1-benzopyran-4-one 500 mg, anhydrous potassium carbonate 2.0 g, 2.0 g of methyl iodide and 30 m of dry acetone are heated under reflux for 15 hours (2 g of methyl iodide are added twice in the middle). The reaction solution is collected by filtration, washed thoroughly with potassium carbonate, and washed acetone is added to the filtrate. The filtrate was concentrated under reduced pressure and the residue was recrystallized from methanol to give 6,7-dimethoxy-2.
400 mg of -phenyl-4H-1-benzopyran-4-one are obtained.

Mp: 192-194 ° C. Elemental analysis: (as C ₁₇ H ₁₄ O ₄₎ Example 3 6,7-Dihydroxy-2-phenyl-4H-1-benzopyran-
800 mg of 4-one 6-hydroxy-7-methoxy-2-phenyl-4H-1-benzopyran-4-one was added to 8 m of hydroiodic acid and 8 mg of acetic anhydride.
Add to the mixture of m and heat to reflux for 3 hours. After the reaction,
Pour the reaction solution into 80 m of ice water. To this mixture is added saturated aqueous sodium hydrogen sulfite solution until the red color of the free iodine disappears. The precipitated crystals were collected by filtration and recrystallized from acetic acid-water,
6,7-Dihydroxy-2-phenyl-4H-1-benzopyran-
300 mg of 4-one monohydrate are obtained.

Melting point: 254-259 ° C Elemental analysis value: (as C ₁₅ H ₁₂ O ₅ ) Example 4 Bone resorption inhibitory effect Bone resorption inhibitory effect "composition culture application research method" page 111-1
14 (Osamu Yamane, Hiroyoshi Endo, Soft Science Publishing,
(1985).

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 ⁴⁵ Ca radioactivity in the collected culture broth is measured with a liquid scintillation counter to convert the ⁴⁵ Ca radioactivity in the whole culture 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 the bone mineral deposited on the cultured bone is determined as the physiological 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 5 Bone formation promoting action 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 one 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 cultivate at 37 ° C in a rotary culture (10 Rotate / time). The compound of the present invention is dissolved in dimethyl sulfoxide, and then directly diluted 1000 times in a culture solution to a concentration of 10 ⁻⁴ molar. While measuring the bone length every day,
Fresh medium is added and bone culture is continued for 6 days. At the end of culturing, the cultured bone is washed with phosphate buffered saline and left in 1N hydrochloric acid for 1 day to elute calcium from the bone tissue, and the amount of Ca eluted is quantified by the chelate method using orthocresolphthalein.

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

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 6 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 6,7-dimethoxy-2-phenyl-4H-1-benzopyran-4-one 300 mg / kg of the present invention and fed for 2 weeks to obtain calcium in the femur and The phosphorus content was measured.

The results are shown below.

Example 7 Acute toxicity 6,7-dimethoxy-2-phenyl-4H-1-benzopyran-4-one was applied to CMC, and 7 weeks old ICR mice each of 10 males and 10 females were used, 1000, 2000, 3000 mg / Oral administration of kg was performed, and observation was performed for 7 days. There were no deaths in any of the groups and no toxic symptoms were observed.

Example 8 Preparation of formulation (a) Tablet 6,7-dimethoxy-2-phenyl-4H-1-benzopyran-4-one
Lactose (95 g) and corn starch (40 g) were mixed with 100 g, and then 5% hydroxypropyl cellulose aqueous solution was added and kneaded, followed by drying. To the dried product, 8 g of carboxymethyl cellulose calcium and calcium stearate were added and mixed, and then 1000 tablets were prepared. Mold.

(b) Capsule 6,7-dimethoxy-2-phenyl-4H-1-benzopyran-4-one
100 g, lactose 39 g and corn starch 35 g are mixed, and 6 g of talc is further 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. , Has a strong bone resorption inhibitory action and a bone formation promoting action, and remarkably inhibits the decrease in the calcium and phosphorus contents in the bones of rats, 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-4H-1-one derivative is useful as a therapeutic agent for osteoporosis.

Claims (4)

[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 or a pharmacologically acceptable salt thereof 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 the following as an active ingredient. 4. 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.