JPH0629183B2 - 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): (In the formula, R ¹ and R ² are each a hydrogen atom or a carbon atom of 1 to
The present invention provides a therapeutic agent for osteoporosis, which comprises a 2-phenyl-4H-1-benzopyran-4-one derivative represented by 3) or a pharmacologically acceptable salt thereof.

[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 never been reported until now that a -4-one derivative (flavone derivative) or a pharmacologically acceptable salt thereof has 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 is by no means satisfactory as a therapeutic agent for osteoporosis. 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 was found that an on derivative (flavone derivative) or a pharmacologically acceptable salt thereof has a strong bone resorption inhibitory action and also 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 shows a strong bone resorption inhibitory action and a bone formation 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 the method described in the literature or a method similar thereto. For example, Organic Synthesis Collective Volume IV, pages 478-481 (Org.Syn.Coll.Vo
l.IV, 478-481); Journal of the Chemical Society (JCS) 1939, 956-960; Proceedings of the Indian Academy of Sciences (Proc. Indian Acad. Sci.) 23 Volume, 2
It can be easily produced by a method such as page 78, (1946) or the like.

That is, 2-phenyl-4H- represented by the general formula (I)
The 1-benzopyran-4-one derivative has the general formula (Wherein R ¹ and R ² have the same meanings as described above), the acetophenone derivative is reacted with benzoyl chloride to give a compound of the general formula (In the formula, R ³ and R ⁴ are a benzoyl group or a carbon number of 1 to
A benzoyloxyacetophenone derivative represented by (an alkyl group of 3)
Baker Venlataraman rearrangemen
t) (Wherein R ³ and R ⁴ have the same meanings as described above), and then dehydration ring closure in the presence of sodium acetate in acetic acid or concentrated sulfuric acid, and hydrolysis if necessary It can be manufactured.

The 2-phenyl-4H-1- represented by the general formula (I) of the present invention
A compound of benzopyran-4-one derivative in which R ¹ and R ² or R ¹ is a hydrogen atom is also prepared by dealkylating a compound in which R ¹ and R ² are lower alkyl groups in the presence of an acid. You can

In particular, in the case of producing a 2-phenyl-4H-1-benzopyran-4-one derivative represented by the general formula (I) in which R ¹ is a hydrogen atom, a 2,6-dihydrooxyacetophenone derivative is used as a raw material. As a tribenzoylmethane derivative is formed in the Baker Bencatalaman rearrangement, the 2-hydroxy-6-alkoxyacetophenone derivative is used as a raw material, and the product is dealkylated in a mixed solution of acetic acid and hydrochloric acid, for example. It is preferable to use a method in which it is produced.

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

In this production method, the acetophenone derivative represented by the general formula (II) used as a raw material 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
The benzopyran-4-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-4H- represented by the general formula (I) of the present invention
The 1-benzopyran-4-one derivative is added to an alcohol solution in which an equivalent amount of sodium hydroxide is dissolved, 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-4-one derivative can be made into a pharmaceutical preparation according to a conventional method. That is, commonly used excipients, disintegrants, binders, pharmaceutical additives such as lubricants are mixed and prepared according to a conventional method to prepare various preparations such as tablets, powders,
It can be a capsule or the like.

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. ~ 100mg is parenterally administered as an injection.

The 2-phenyl-4H-1- represented by the general formula (I) of the present invention
Benzopyran-4-one derivatives or their pharmacologically acceptable salts showed strong bone resorption inhibitory action and osteogenesis promoting action in in vitro experiments using chicken embryo femur, and when they were fed a calcium-deficient diet. It has an action of remarkably suppressing the decrease in the content of calcium and phosphorus in the bones of the resulting rat, and is useful as a highly safe 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-5-methoxy-2-phenyl-4H-1-benzopyran-4-one (a) 3,6-dibenzoyloxy-2-methoxyacetophenone 3,6-dihydroxy-2- Methoxy acetophenone 1.1g
Was dissolved in 3m of dry pyridine and benzoyl chloride 1.80g
Is added and the mixture is heated on a boiling water bath for 20 minutes, then ice is added to the reaction solution and the pH is adjusted to 1 with 10% hydrochloric acid. The precipitated crystals are collected by filtration, washed with water and recrystallized from ethanol to obtain 2.1 g of pale brown needle crystals 3,6-dibenzoyloxy-2-methoxyacetophenone.

Melting point: 158-159 ° C (b) 3-benzoyloxy-6-hydroxy-2-methoxydibenzoylmethane 3,6-dibenzoyloxy-2-methoxyacetophenone 7.7
g was dissolved in 35 m of dry N, N-dimethylformamide, 1.03 g of 60% sodium hydride (oil) was added to this solution over 5 minutes, and the mixture was left at room temperature for 15 minutes. The reaction solution is poured into 200 m of water and adjusted to pH 1 with 10% hydrochloric acid. The precipitated crystals are collected by filtration, washed with water and then recrystallized from ethanol to obtain 5.6 g of yellow crystals of 3-benzoyloxy-6-hydroxy-2-methoxydibenzoylmethane.

Melting point: 154-155 ° C (c) 6-Hydroxy-5-methoxy-2-phenyl-4H-1-benzopyran-4-one 3-benzoyloxy-6-hydroxy-2-methoxydibenzoylmethane 23.1 g Add 240 ml of concentrated sulfuric acid to completely dissolve it, and then stir at room temperature for 10 minutes. 2 reaction solutions
It is poured into ice water, and the precipitated crystals are collected by filtration and washed with water. The precipitated crystals are recrystallized from ethanol-water to obtain 12.8 g of pale yellow crystals of 6-hydroxy-5-methoxy-2-phenyl-4H-1-benzopyran-4-one.

Melting point: 189.5 to 190 ° C Elemental analysis value: (as C ₁₆ H ₁₂ O ₄ ) Example 2 5,6-dimethoxy-2-phenyl-4H-1-benzopyran-4-one 6-hydroxy-5-methoxy-2-phenyl-4H-1-benzopyran-4-one 4.55 g was dried with 300 m of acetone. Dissolves in
Add 20 g of methyl iodide and 25 g of anhydrous potassium carbonate.
The mixture is heated at reflux for 6.5 hours. The reaction solution is filtered,
Water is added to the filtrate, and acetone is distilled off under reduced pressure. The precipitated crystals are collected by filtration, washed with water and recrystallized from ethanol to give colorless crystals of 5,6-dimethoxy-2-phenyl-4H-1-benzopyran-4-one (3.95 g).

Melting point: 201-202.5 ° C Example 3 5,6-dihydroxy-2-phenyl-4H-1-benzopyran-
4-one 6-hydroxy-5-methoxy-4H-1-benzopyran-4-one 1.0 g, acetic acid 30 m and concentrated hydrochloric acid 30 m were mixed with 2.5
Heat to reflux for hours. The reaction solution was poured into ice water, and the precipitated crystals were collected by filtration, washed with water, and recrystallized from ethanol-water to give yellow crystals of 5,6-dihydroxy-2-phenyl-4H-1-benzopyran-4-one 0.63. g is obtained.

Melting point: 189.5 to 190.5 ° C Elemental analysis value: Elemental analysis value: (as C ₁₅ H ₁₀ O ₄ ) Example 4 5-Hydroxy-6-methoxy-2-phenyl-4H-1-benzopyran-4-one 5,6-dimethoxy-2-phenyl-4H-1-benzopyran-4-one
A mixture of 470 mg, 20 m acetic acid and 20 m concentrated hydrochloric acid is heated to reflux for 2 hours. Ice water was added to the reaction solution, and the precipitated crystals were collected by filtration and recrystallized from ethanol to give yellow crystals of 5-hydroxy-6-methoxy-2-phenyl-4H-1-benzopyran-4-one 3
You get 50 mg.

Melting point: 129.5 to 131 ° C Elemental analysis value: (as C ₁₆ H ₁₂ O ₄ ) Example 5 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 on 10 to 11 days of incubation and removing the soft tissues attached to the bones well, the phenol red containing the 2-phenyl-4H-1-benzopyran-4-one derivative of the present invention was added. Not BGJb-HW2 culture medium (hereinafter referred to as culture medium) 1 m
Preculture is carried out at 37 ° C. for 1 day by using the rotating 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 from the culture solution every 12, 24, 48, and 72 hours, discard the remaining culture solution, and add a new culture solution. ⁴⁵ in the collected culture
The Ca radioactivity is measured with a liquid scintillation counter to calculate the radioactivity of ⁴⁵ Ca in the whole culture solution. 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 linearly regressed, 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. The efficacy of the compound of the present invention for inhibiting bone resorption is determined by the following formula using the value of T1 / 2.

The results are shown below.

Example 6 Bone formation promoting action Bone formation promoting action is shown in "Tissue culture applied research method" pages 103 to 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-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.

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.

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 7 Effect on Calcium and Phosphorus Contents in Bone of Rats Fed a Ca-Deficient Diet 20 Wistar male rats aged 3 weeks were used in groups of 10 2
The group is divided into groups, a Ca-deficient diet in one group, a Ca-deficient diet in the other group, and 6-hydroxy-5-methoxy-2-phenyl-4H-1-of the present invention.
Benzopyran-4-one 300 mg / kg was given and raised for 2 weeks,
The amount of calcium and phosphorus in the femur was measured.

The results are shown below.

Example 8 Acute toxicity 6-Hydroxy-5-methoxy-2-phenyl-4H-1-benzopyran-4-one was applied to CMC, and 10-week-old ICR mice of 10 males and 10 females were used. , 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 9 Preparation of preparation (a) Tablets 6-Hydroxy-5-methoxy-2-phenyl-4H-1-benzopyran-4-one 100 g, lactose 95 g and corn starch 40 g are mixed and then 5% hydroxypropyl An aqueous cellulose solution is added and kneaded, followed by drying. 8 g of carboxymethylcellulose calcium and calcium stearate are added to the dried product and mixed, and then molded into 1000 tablets.

(b) Capsules 6-Hydroxy-5-methoxy-2-phenyl-4H-1-benzopyran-4-one (100 g), lactose (39 g) and corn starch (35 g) were mixed, and then 6 g of talc was added to the mixture and mixed. Fill 1000 hard capsules.

〔The invention's effect〕

The 2-phenyl-4H-1-benzopyran-4-one derivative represented by the general formula (I) and the pharmacologically acceptable salt thereof of the present invention can be used in in vitro experiments using chicken embryo femurs.
It has a strong bone resorption inhibitory action and a bone formation promoting action, and remarkably suppresses 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-4-one derivative is useful as a therapeutic agent for osteoporosis.

Claims (5)

[Claims] 1. A general formula (In the formula, R ¹ and R ² are each a hydrogen atom or a carbon atom of 1 to
A 2-phenyl-4H-1-benzopyran-4-one derivative represented by the formula (3 which is an alkyl group of 3) or a pharmacologically acceptable salt thereof 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 or a pharmacologically acceptable salt thereof 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 the following as an active ingredient. 5. 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.