JPH09188665A - New substituted n-quinolylanthranyl acid redivative and production of the same - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain the subject new compound having a potent accelerating action on a bone formation by osteoblastic cells, and useful for the treatment of a metabolic bone disease. SOLUTION: This new substituted quinolylanthranylic acid is a compound of formula I (when R<1> is F, R<2> is H or a halogen; R<3> is H and when R<1> , R<2> are both H, R<3> is an acetyl or a 3-5C alkyl; R<4> is H, a lower alkyl or 2,3- dihydroxypropyl), e.g. N-(6-fluoro-4-quinolyl) anthranylic acid 2, 3- dihydroxypropyl ester. The compound of the formula I is obtained by reacting an anthranylic acid derivative of formula II with a quinoline derivative of formula III to obtain a corresponding ester body, and de-halogenating or hydrolyzing as necessary. The compound of the formula I has only the weak action of a worried cyclooxygenase inhibition and a high safety, and can increase a bone mass and a bone strength for reducing a pain such as a lumbago and the risk of a bone fracture in a metabolic bone disease such as an osteoporosis.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a novel N-quinolylanthranilic acid derivative or a pharmaceutically acceptable salt, and a therapeutic agent for metabolic bone disease containing these as active ingredients.

[0002]

2. Description of the Related Art In normal bone metabolism, homeostasis is maintained because the amount of bone resorption by osteoclasts and the amount of bone formation by osteoblasts are in equilibrium. It is thought that metabolic bone disease occurs when the balance between bone resorption and bone formation is disrupted.
This disease includes osteoporosis, fibro-osteitis (hyperparathyroidism), osteomalacia, and Paget's disease, which affects systemic bone metabolism parameters. In particular, osteoporosis is one of the geriatric diseases, and it is common in postmenopausal women. Symptoms include pain such as low back pain and fractures, and particularly, fractures of the femoral neck are serious because they cause debilitating and dementia of the whole body.

For the treatment and prevention of such bone diseases, calcium preparations, active vitamin D ₃ preparations, calcitonin preparations and estrogen preparations are used.

[0004]

Many of the above-mentioned conventional therapeutic agents have been reported to have an action of suppressing bone resorption, but none have clearly shown an action of promoting bone formation.
In addition, the subject to be administered may be limited or the effect may be uncertain, so that the sufficient effect is not obtained at present.

From the above, in order to reduce the risk of pain such as low back pain and the risk of fracture in metabolic bone diseases such as osteoporosis, it is necessary to increase the bone mass and bone strength, and the effect is more certain. There is a strong demand for the development of a drug having an action of promoting bone formation by possible osteoblasts.

Further, N- having different substituents from the compound of the present invention
As quinolylanthranilic acid derivatives, those having anti-inflammatory and antipyretic analgesic action are described in U.S. Pat.No. 3,376,195 (1968) and Chemical Abstracts, Volume 80, 51u (1973). Is not touched at all.

[0007]

Means for Solving the Problems The inventors of the present invention have conducted diligent research for the purpose of developing a therapeutic drug having an osteogenesis promoting action by promoting the function of osteoblasts. As a result, the following general formula (1) It has been confirmed that the N-quinolylanthranilic acid derivative represented by the formula or a pharmaceutically acceptable salt thereof has a strong osteogenesis promoting action by osteoblasts, and further has a weak cyclooxygenase inhibitory action, which is highly safe. The present inventors have completed the present invention by discovering that it can be an excellent therapeutic drug for metabolic bone disease. [In the formula, when R ¹ is a fluorine atom, R ² is hydrogen or a halogen atom, R ³ is hydrogen, and when R ¹ and R ² are both hydrogen, R ³ is an acetyl group and has 3 to 5 carbon atoms. Represents an alkyl group, R ⁴ represents hydrogen, a lower alkyl group or a 2,3-dihydroxypropyl group]

N- represented by the above general formula (1) of the present invention
The quinolyl anthranilic acid derivative can be produced as follows. For example, general formula (2) An anthranilic acid derivative represented by the formula [wherein R ⁴ represents hydrogen, a lower alkyl group or a 2,3-dihydroxypropyl group] and the general formula (3) [In the formula, when R ¹ is a fluorine atom, R ² is hydrogen or a halogen atom, R ³ is hydrogen, and when R ¹ and R ² are both hydrogen, R ³ is an acetyl group and has 3 to 5 carbon atoms. An alkyl group and R ⁵ represents a halogen atom] in a solvent-free or inert solvent, preferably 0.5 to 1
It can be produced by heating and stirring in N diluted hydrochloric acid, and if necessary, dehalogenation by catalytic reduction using a metal catalyst, preferably palladium carbon, or hydrolysis in the presence of an acid or a base.

Here, "lower alkyl" refers to an alkyl group having 1 to 3 carbon atoms such as methyl, ethyl, propyl and isopropyl, and "halogen atom" refers to a fluorine, chlorine, bromine or iodine atom. Alkyl having 3 to 5 carbon atoms "
Is a linear, branched or unsaturated alkyl group such as isopropyl, butyl, tertiary butyl, pentyl and the like.

In the present production method, the compounds represented by the above general formula (2) used as raw materials are all known compounds, and are available as commercial products, or produced by a method described in literature or a method similar thereto. be able to. Further, the compound represented by the general formula (3) can be produced by the method disclosed in Chemical Abstracts, Volume 80, 51u (1973).

Further, the compound represented by the general formula (1) of the present invention can be converted into a pharmaceutically acceptable salt according to a conventional method. Examples of such salts include inorganic bases such as sodium, potassium, magnesium, calcium and aluminum, organic bases such as methylamine, ethylamine and ethanolamine.

[0012]

EXAMPLES The following examples are given to further illustrate the present invention. All melting points are measured by the hot plate method and are uncorrected.

Example 1 N- (6-fluoro-4-quinolyl) anthranilic acid
2,3-dihydroxypropyl ester

2.93 g glycerol, sodium hydroxide
A mixture of 0.157 g and 0.510 g of isatoic anhydride was slowly heated to 60 ° C. with stirring, further heated to 80 ° C. and stirred with heating for 1 hour. 4-chloro-6-fluoroquinoline 0.201
g, 6.7 ml of 0.5N hydrochloric acid was added, and the mixture was further stirred with heating at 80 ° C. for 2 hours. The reaction solution was added to ice water, made alkaline with potassium carbonate, and the precipitated crystals were collected by filtration. The crystals obtained were dried by heating and then recrystallized from chloroform to give white powder crystals.
0.203 g of the desired product was obtained. Melting point 173-174 ° C

[0015]

Example 2 Ethyl N- (7-chloro-6-fluoro-4-quinolyl) anthranilate

4,7-dichloro-6-fluoroquinoline
1.13g, ethyl anthranilate 0.956g, 0.5N hydrochloric acid 15m
The mixture of 1 was heated and stirred at 80 ° C. for 4 hours. After cooling, it was neutralized with aqueous sodium hydrogen carbonate and extracted with methylene chloride. The organic layer was washed successively with water and saturated brine, dried over sodium sulfate, and the solvent was evaporated under reduced pressure. The obtained crystals were recrystallized from ethanol to give 0.910 g of the desired product as pale yellow needle crystals. Mass spectrum: m / z
344 (M ⁺ ), melting point 167.5-169 ° C

[Examples 3 and 4] In the same manner as in Example 2, the compounds shown in Table 1 were obtained.

[0019]

[Table 1]

Example 5 Ethyl N- (6-fluoro-4-quinolyl) anthranilate

N- (7-chloro-6 obtained in Example 2
Ethyl (fluoro-4-quinolyl) anthranilate 0.5
A mixture of 02 g, 10% palladium on carbon (0.100 g) and ethanol (70 ml) was reduced under hydrogen pressure of 3.0 kg / cm ² at room temperature for about 5 hours. The insoluble material was filtered off, and the filtrate was concentrated under reduced pressure to obtain 0.530 g of the desired product as a yellow oily substance. Mass spectrum: m / z 311
[M + H] ⁺

Example 6 N- (6-fluoro-4-quinolyl) anthranilic acid

N- (6-fluoro-obtained in Example 5
Ethyl 4-quinolyl) anthranilate 0.447 g, 2.53M
A mixture of 5 ml of an aqueous sodium hydroxide solution and 10 ml of methanol was heated and stirred at 80 ° C. for about 3 hours. After cooling, the mixture was made weakly acidic with acetic acid, the precipitated crystals were collected by filtration, and the obtained crystals were washed with water, ethanol and methylene chloride successively and dried to give 0.402 g of the desired product as yellow prism crystals. Melting point: 307 ~ 308 ℃

[0024]

[Examples 7 to 9] The compounds shown in Table 2 were obtained in the same manner as in Example 6.

[0026]

[Table 2]

[Experimental Example 1] Bone formation promoting action As phenotypes of osteoblasts, alkaline phosphatase (ALP-ase) activity, production of bone matrix proteins (collagen, osteocalcin, osteonectin, osteopontin, etc.), active vitamin The presence of D ₃ receptor, parathyroid hormone receptor, estrogen receptor, androgen receptor can be mentioned (Molecular
Medicine, Vol. 30, No. 10, 1232 (1993)). ALP-ase
Is higher than the initial stage of functional expression of osteoblasts (Journal
Of Cellular Physiology, Volume 143, 420 (199
0)). It is said that the role of ALP-ase on osteogenesis by osteoblasts is to increase the concentration of phosphate ion in the bone formation site and to decompose pyrophosphate which is a mineralization inhibitor (cell engineering, 13 Volume, Issue 12, 1062 (199
Four)). Moreover, it has been shown that calcification occurs when subcutaneously transplanting a substance in which ALP-ase and a collagen sheet are covalently bound (Journal of Clinical
Investigation, Vol. 89, 1974 (1992)). Therefore, it is possible to use the increase in ALP-ase activity of osteoblasts as an evaluation system for promoting osteogenesis. Therefore, the osteogenesis promoting action of the compound of the present invention was evaluated using the increase in ALP-ase activity of osteoblasts as an index. The test methods and test results are listed below.

The mouse osteoblast cell line was seeded in a 6-well plate at 5 × 10 ⁴ / well, and α containing 10% fetal bovine serum was used.
-Cultivated in MEM medium. After 6 days, the test compound was added and the cells were cultured for 4 days in an α-MEM medium containing 1% fetal bovine serum. The cell layer was washed with PBS (-) (phosphate buffered saline), 0.2% Nonidet P-40 containing 1 mM magnesium chloride was added, and the cells were sonicated to solubilize them. After centrifuging this at 3000 rpm at 4 ° C for 10 minutes, the ALP-ase activity and the protein amount in the supernatant were measured. ALP-
The ase activity was determined by measuring the amount of p-nitrophenol released by enzymatic reaction at 37 ° C. for 30 minutes using p-nitrophenyl phosphate as a substrate by measuring the absorbance at OD _{410 nm} . The protein mass is determined by the protein assay (Bio-Ra
d company).

The ALP-ase activity value was calculated according to the following formula.

Table 3 shows the measurement results of the example compounds.

[0031]

[Table 3]

[Experimental Example 2] Measurement of cyclooxygenase inhibitory action The action on cyclooxygenase activity was evaluated using the production of malondialdehyde (MDA) in rabbit platelets as an index. Platelet-rich plasma was prepared from blood containing 0.38% sodium citrate of Japanese white male rabbits. 15 mM Tris-
1.1 × 10 ⁹ platelets / in hydrochloric acid buffered saline (pH 7.4)
It was suspended so that it would be ml. 0.1 ml of the compound solution was added to 0.9 ml of the platelet suspension, and the mixture was incubated at 37 ° C for 5 minutes. 20 μl of 2.5 mM sodium arachidonate was added, and the mixture was incubated at 37 ° C. for 3 minutes. The reaction was stopped by adding 1 ml of 0.8% 2-thiobarbituric acid aqueous solution. After boiling at 100 ° C for 10 minutes, it was quenched with running water. After centrifugation at 3000 rpm for 10 minutes, the absorbance of the supernatant at 532 nm was measured. The amount of MDA produced was determined as nmol / 10 ⁹ platelets from the calibration curve. The cyclooxygenase activity inhibition rate was calculated from the following formula.

[0033]

[0034]

[Table 4]

Claims (4)

[Claims] 1. General formula (1) [In the formula, when R ¹ is a fluorine atom, R ² is hydrogen or a halogen atom, R ³ is hydrogen, and when R ¹ and R ² are both hydrogen, R ³ is an acetyl group and has 3 to 5 carbon atoms. An alkyl group and R ⁴ represents hydrogen, a lower alkyl group or a 2,3-dihydroxypropyl group], or an N-quinolylanthranilic acid derivative or a pharmaceutically acceptable salt thereof. 2. General formula (1) [In the formula, when R ¹ is a fluorine atom, R ² is hydrogen or a halogen atom, R ³ is hydrogen, and when R ¹ and R ² are both hydrogen, R ³ is an acetyl group and has 3 to 5 carbon atoms. An alkyl group and R ⁴ represents hydrogen, a lower alkyl group or a 2,3-dihydroxypropyl group], or an pharmaceutically acceptable salt as an active ingredient. A therapeutic agent for metabolic bone disease, which is characterized in that 3. An N-quinolylanthranilic acid derivative and a pharmaceutically acceptable salt according to claim 1, wherein the compound is N- (6-fluoro-4-quinolyl) anthranilic acid 2,3-dihydroxypropyl ester. . 4. The general formula (2) An anthranilic acid derivative represented by the formula [wherein R ⁴ represents hydrogen, a lower alkyl group or a 2,3-dihydroxypropyl group] and the general formula (3) [In the formula, when R ¹ is a fluorine atom, R ² is hydrogen or a halogen atom, R ³ is hydrogen, and when R ¹ and R ² are both hydrogen, R ³ is an acetyl group and has 3 to 5 carbon atoms. An alkyl group and R ⁵ represents a halogen atom] is reacted to produce a corresponding ester, and if necessary dehalogenation or hydrolysis is carried out by the general formula (1 ) A method for producing an N-quinolylanthranilic acid derivative or a pharmaceutically acceptable salt represented by the formula [wherein R ¹ , R ² , R ³ and R ⁴ are as described above].