JPS63297325A - Remedy for osteoporosis - Google Patents

Abstract

PURPOSE:To obtain a remedy for osteoporosis, having inhibitory action on bone resorption, promoting action on bone formation and high safety, containing a benzofuro[3,2-c]quinoline derivative or a salt thereof as an active ingredient. CONSTITUTION:A benzofuro[3,2-c]quinoline derivative shown by the formula (R is H or OH) or a salt thereof is blended with a conventional drug carrier and pharmaceutically manufactured by a conventional procedure to give the aimed substance. The substance can be processed into a dosage form such as tablet, powder, capsule or injection. A dose is 10-1,000mg per adult daily in the case of oral administration and 1-100mg per adult daily in the case of parenteral administration. The compound shown by the formula is obtained by reacting m-anisidine with a phenylmalonic acid derivative, treating the reac tion product with pyridine hydrochloride and dealkylating simultaneously with ring closure. 3-Hydroxy-5H-benzofuro[3,2-c]quinolin-6-one may be cited as the compound shown by the formula.

Description

DETAILED DESCRIPTION OF THE INVENTION [Industrial Application Field] The object of the present invention is to provide benzofurol"3.2-c represented by the general formula (I) (R in the formula is a hydrogen atom or a hydroxyl group) ) A therapeutic agent for osteoporosis containing a quinoline derivative or a pharmacologically acceptable salt thereof is provided.

[Conventional technology]

Osteoporosis is a pathological condition in which bone mass is decreased without any change in the chemical composition of bones, and its physiological characteristic is a decrease in protein, calcium, and phosphorus in bones.

Osteoporosis increases with age and usually affects the spinal cord, causing back pain and shortened height. In particularly advanced cases, long bones are also affected, sometimes resulting in fractures.

It is said that most of the causes of femur fractures seen in the elderly are due to senile osteoporosis.

There are various causes of this osteoporosis, including endocrine and nutritional disorders, but vitamin D preparations, calcium preparations, calcitonin preparations, phosphorus preparations, etc. that have been used as therapeutic agents for osteoporosis have not been targeted. Since the effects are limited and the effects are uncertain, there is a strong desire to develop formulations with more reliable efficacy.

In recent years, a certain type of drug with a completely different chemical structure from the above-mentioned preparations has been developed.
It has been reported that -phenyl-4H-1-benzopyran-4-one derivatives have a bone resorption inhibitory effect and are useful as therapeutic agents for osteoporosis (Japanese Patent Publication No. 13391/1983).
JP-A-60-48924, JP-A No. 60-54379, JP-A No. 60-132917, JP-A No. 60-132976)
.

As the benzofuro(3,2-c)quinoline derivative of the present invention, the formula or the compound represented by the formula has already been synthesized and reported by Youse et al. [Pretin on the Chemical Society on Japan (Bull.

Chem, Soc, Jpn, > 53 volumes, 1057~
1060 pages.

1980; Journal on Heterocyclic Chemistry (J, Heterocyc
lic Chem,) Volume 21.

pp. 737-739, 1984].

However, all of these were synthesized for synthetic interest or to confirm chemical reactivity, and in terms of pharmacological activity, they may have mutagenic, carcinogenic, or anticancer effects. It is only shown that there is no explanation of its effect. Furthermore, as in the present invention, there has been no report whatsoever that benzofuro(3,2-C)quinoline derivatives exhibit a bone resorption inhibitory effect and are useful as therapeutic agents for osteoporosis.

[Problem that the invention seeks to solve]

3-phenyl-411- disclosed in said patent application
1-benzopyran-4-one derivatives have a weak bone resorption inhibitory effect and are by no means satisfactory as therapeutic agents for osteoporosis. Therefore, the present inventors conducted intensive studies to find compounds with stronger bone resorption inhibitory effects, and found that certain benzofuro(3,2-c)quinoline derivatives or their pharmacologically acceptable salts have a stronger effect. It has been found that it has an effect of inhibiting bone resorption and also an effect of promoting bone formation, and can be an excellent therapeutic agent for osteoporosis.

The present invention is based on these findings.

[Means for solving problems]

Benzofuro [3,
2-C) Quinoline derivatives or their pharmacologically acceptable salts exhibit strong bone resorption inhibitory and bone formation promoting effects and are useful as highly safe osteoporosis therapeutic agents.

Benzofuro (3,
2-C) Some of the quinoline derivatives include new compounds, but all of them can be prepared using methods described in the literature, such as pretin ion.
The Chemical Society Japan (Bul
l, Chem, Soc, Jpn,) Volume 53 1
Pages 057-1060 (1980); Journal on Heterocyclic Chemistry (J, 1et
Ero-cyclic Chem,) Volume 21,
737-739 (1984) or similar methods thereof.

For example, m-anisidine represented by the formula and -AQ formula (Ro in the formula
is a hydrogen atom or a lower alkoxy group) to produce a compound represented by the general formula (R" in the formula has the same meaning as above), and It can be produced by treating with pyridine hydrochloride to simultaneously dealkylate and ring close.

In this production method, the general formula (
The compound represented by III) can be prepared by methods described in the literature, for example,
Journal on Heterocyclic Chemistry (J, Heterocyclic Ch.
em,) Volume 21.

It can be produced according to the method described on pages 737 to 739 (1984) or a method similar thereto.

Benzofuro [3,
2-c) The quinoline derivative can be converted into a pharmacologically acceptable salt according to a conventional method. For example, the benzofuro[3゜2-〇]quinoline derivative represented by the general formula (I) of the present invention is added to an alcoholic solution containing an equivalent amount of sodium hydroxide, heated, and then concentrated under reduced pressure. By doing so, the sodium salt can be obtained.

Benzofuro [3,
2-c) The quinoline derivative can be made into various pharmaceutical preparations according to conventional methods. That is, by mixing with pharmaceutical additives such as excipients, disintegrants, binders, and lubricants as necessary, and preparing according to conventional methods, various preparations such as tablets, powders, capsules, etc. are prepared. be able to.

Benzofuro (3,
2-C) When using a quinoline derivative as a therapeutic agent for osteoporosis, approximately 10 to 1000 mg per day for adults is prepared into an appropriate dosage form, such as tablets, powders, capsules, etc., and administered orally, or approximately 1000 mg per day for adults is administered. 1 to 100 mg is administered parenterally in the form of an injection or the like.

Benzofuro [3,
2-C) Quinoline derivatives or their pharmacologically acceptable salts were tested in vitro using chicken embryo femurs.
It exhibits strong bone resorption inhibitory and bone formation promoting effects1, and has the effect of significantly suppressing the decrease in calcium and phosphorus content in the bones of rats that occur when fed a calcium-deficient diet, and has low toxicity. , is useful as a highly safe osteoporosis treatment agent.

〔Example〕

Examples are given below to further explain the present invention.

Note that all melting points of compounds in each example are uncorrected.

Example 1 2.7 g of m-anisidine and 5.32 g of diethyl 2-methoxyphenylmalonate were added to 20 ml of diphenyl ether.
2 and heated at 270 to 290°C for about 2.5 hours with an air cooling tube attached. After cooling, diethyl ether 807 was added and the precipitated crystals were collected by filtration and washed with diethyl ether to give 5.39 g (90,') of 4-hydroxy-7-medoxy-3-(2-methoxyphenyl)-2-quinolone.
7%).

Melting point: >300°C IR (KBr): νen 1620 cm-
'NMR (d6-DMSO) δ: 3.69 (s, 3H), 3.81 (s, 3
H), 6.75-6.79 (m, 2H), 6.9
3-7.13 (m, 3H), 7.29-7.35 (
m, IH), 7.76-7.80 (m, 1B).

9, '56 (br-s, IH), 11.18 (s,
Lfl) Elemental analysis value' (as C+J+sNL) 6% 8% N% Calculated value 68.68 5.09 4.71 Actual value 6g, 79 5.08 4.724-
A mixture of 5 g of hydroxy-7-medoxy-3-(2-methoxyphenyl)-2-quinolone and 50 g of pyridine hydrochloride was heated under reflux at 220-250° C. for 2.5-3 hours. Pour the reaction mixture onto 200-300 g of hot crushed ice,
The precipitated crystals were collected by filtration, washed with water, and recrystallized with alcohol to obtain 1.9 g of 3-hydroxy-58-bensif o[3,2-c)quinolin-6-one.

Melting point: >300°C IR (KBr): νCo 1640 Cm
-'NMR (d6-DMSO) δ: 6.92~7.04 (m, 2N), 7.52~
7.60 (m.

2H), 7.89-8.17 (m, 3H),
10.49 (s.

It (), 11.88 (s, IH) elemental analysis value: (C+5l (as sNO3) 0% 8%
N% Calculated value 71.71 3.61 5.58 Actual value 71.37 3.61 5.44 Example 2 Instead of using diethyl 2-methoxyphenylmalonate, diethyl 2.4-dimethoxyphenylmalonate or 2.4-dimethoxyphenylmalonate was used. The following compounds were synthesized in the same manner as in Example 1 except that diethyl 5-dimethoxyphenylmalonate was used.

Melting point: >300℃ IR (KBr): νco 1640 am
-'NMR (d, -DMSO) δ: 6.77-6.91 (m, 3H), 7
．． 11-7.12 (m.

IH), 7.78-8.30 (m, 2H),
9.83 (s.

IH), 10.22 (s, 1fl), 11
．． 67 (s, IH) elemental analysis value: (CISH9
(as NO4) 0% 8% N% Calculated value 67.42 3.39 5.24 Actual value 67.2m 3.41 5.35
Point: >300℃ IR (KBr): L”.o 1660.16
30 Cm-'NMR (d6-DMSO) δ: 6,77-6.90 (m, :E), 7.39
~7.85 (m.

3H), 9.46(s, 1N), 10.31(s
, 1tl).

11.64 (s, LH) Elemental analysis value: (as CISH9NO4) 0%
8% N% Calculated value 67.42 3.39 5.24 Actual value 67.71 3.45 5.54 Example 3 Bone resorption inhibitory effect Bone resorption inhibitory effect was evaluated in "Tissue Culture Applied Research Methods" 111-11
The measurement was performed according to the method described on page 4 (edited by Satoshi Yamane and Hiroyoshi Endo, published by Soft Science Publishing, 1985). After extracting the 10-11-day-old chicken embryo femur and thoroughly removing the soft tissue attached to the bone, the benzofuro of the present invention (3,
2-C) Preculture is performed using 1 mi2 of phenol red-free BGJb-)IW2 culture solution (hereinafter referred to as culture solution) to which a quinoline derivative has been added at 37° C. for 1 day by the rotation culture method. The compound of the present invention is first dissolved in dimethyl sulfoxide to prepare a 0.1 molar solution, which is diluted 1000 times with a culture medium to give a 10-4 molar concentration. In addition, a control group is cultured with the same volume of dimethyl sulfoxide alone added.

The next day, add 1 μCi/mA of 45CaCI2 to the fresh culture medium.
The chicken embryo femur, which had been dissolved and precultured at a concentration of
and culture with shaking at 37°C for 2 hours. As a result, the bone mineral in the cultured bone is labeled with 4SCa. Immediately after culturing, the cultured bone was washed with phosphate buffered saline that had been preheated to 37°C to remove the 4S that adhered to the bone.
(Remove a.) This 4SCa labeled cultured bone is again cultured in a culture medium using the rotation culture method (at the time of 10 rotations).

12. 24. 48. Every 72 hours, take an exact amount of culture fluid from the culture fluid, and at the same time discard the remaining culture fluid and add fresh culture fluid. 4S Ca in the collected culture solution
Radioactivity was measured using a liquid scintillation counter.
Calculate the radioactivity of 45 (a) in the total culture solution. After the completion of the culture, the bone tissue was left in 1N hydrochloric acid for 1 day to elute the total calcium, and its radioactivity was measured. Define as final residual radioactivity.

From the obtained measurement values, the ratio of the radioactivity remaining in the cultured bone to the total radioactivity initially incorporated into the bone tissue was calculated, and the decay curve of the residual radioactivity in the cultured bone after 24 hours was calculated. Bone mineral elution by osteocytes is linearly regressed, and from the slope of the obtained straight line, the turnover rate of calcium in the bone mineral deposited on the cultured bone is determined by the biological half-life T%.

The compound group of the present invention and the control group were each conducted with 5 patients per group.

Compared with the T'A value of the control group, the T'A value of the compound group of the present invention
'A large value of A indicates that the compound of the present invention has a bone resorption inhibitory effect. The efficacy of the bone resorption inhibitory effect of the compound of the present invention is determined by the following formula using the value of T'A.

The results are shown below.

[Compound] [Efficacy of bone resorption inhibitory action] Compound 1 1.24 Compound 2 1.97 Compound 32.52 Example 4 Osteogenesis-promoting effect Osteogenesis-promoting effect was observed in "Tissue Culture Applied Research Methods" 103-11
The measurement was performed according to the method described on page 1 (edited by Satoshi Yamane and Hiroyoshi Endo, published by Soft Science Publishing, 1985).

The femur bones of 9-day-old chicken embryos were removed, the soft tissues attached to the bones were thoroughly removed, and one of the left and right femurs of each individual was used as the compound group of the present invention and the other as the control group, and a rectangular culture plate was prepared. Attach it one by one to the inner surface of the test tube, and add BGJb-HW to it.
Add 2 ml of 2 culture medium solution (hereinafter referred to as culture solution), seal with a silicone stopper, and culture with rotation at 37°C (10 rotations).

The compound of the present invention is once dissolved in dimethyl sulfoxide to prepare a 0.01 molar solution, which is diluted 1000 times with a culture medium to a 10-5 molar concentration. In addition, a control group is cultured with the same volume of dimethyl sulfoxide alone added.

Preculture is continued for 6 days while measuring bone length every day and replacing with fresh culture medium.

At the end of the culture, wash the cultured bone with phosphate buffered saline,
The bone tissue is left in normal hydrochloric acid for one day to elute calcium from the bone tissue, and the amount of eluted Ca is determined by the chelation method using orthocresol phthalein.

This experiment was conducted with 6 patients in each group.

The efficacy of the osteogenesis promoting effect of the compound of the present invention was determined using the following formula.

The results are shown below.

Compound Compound with osteogenesis promoting effect 2
1.11 Example 5 Twenty 3-week-old Wistar male rats were divided into two groups, each group containing 10 rats, and one group was treated with the test compound 3,9-dihydroxy-5-day-benzofuro (3,2-C). Quinolin-6-one (compound 2) 300 kg/kg was administered orally once a day as a CMC suspension by force, and the other group received the same amount of CMC alone. The mice were fed a Ca-deficient diet and reared for two weeks, and the amounts of calcium and phosphorus in the femur were measured.

The results are shown below.

Example 6 Acute toxicity 3.9-dihydroxy-5-day-benzofuro[3,2-C
) Quinolin-6-one was suspended in CMC, and 7-week-old I
Using 10 male and 10 male and female cR mice, 1000.2000
．． 3000 mg/kg was orally administered and observed for 7 days. There were no deaths in either group, and no pre-symptomatic symptoms were observed.

Example 7 Manufacture of formulation (a) Tablet 3.9-dihydroxy-50-benzofuro (3,2-C
:]] Mix 100 g of quinolin-6-one, 95 g of lactose and 40 g of corn starch, then 5%
After adding and kneading an aqueous solution of hydroxybrobyl cellulose, drying and adding 8 g of calcium carboxymethylcellulose and 7 g of calcium stearate to the dried product.
After adding and mixing, form into 1000 tablets.

(b) Capsules 3,9-dihydroxy-5-day-benzofuro (3,2-C
) 100 g of quinolin-6-one, 59 g of lactose and 35 g of corn starch were mixed, and 6 g of talc was added to the mixture and mixed, followed by 1000 hard capsules.
Fill into capsules.

〔Effect of the invention〕

Benzofuro (3,2-
C] Quinoline derivatives and their pharmacologically acceptable salts were tested in an in vitro experiment using chicken embryo femur.
It exhibits significant bone resorption inhibiting and bone formation promoting effects at -4 to 10-5 molar concentrations. Also, 1000-3000 mg
/ kg was orally administered, there were no deaths and no serious poisoning symptoms were observed.

Therefore, benzofuro (
3,2-C:]quinoline derivatives are extremely useful compounds as therapeutic agents for osteoporosis.

patent applicant Kinosei Pharmaceutical Industry Co., Ltd.

Claims (1)

[Claims] Benzofuro[3,2-c]quinoline derivatives represented by the general formula ▲ Numerical formulas, chemical formulas, tables, etc. ▼ (R in the formula is a hydrogen atom or a hydroxyl group) or their pharmacology A therapeutic agent for osteoporosis containing a sexually acceptable salt as an active ingredient.