JP2003119142A - Agent for inhibiting formation of advanced glycation end product - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide an agent for inhibiting the formation of advanced glycation end product (AGE). SOLUTION: The AGE-formation inhibiting agent contains a 1,4- benzothiazine-2-acetic acid derivative of general formula (I) (R<1> and R<2> are each H, a halogen atom, a lower alkyl, a lower alkoxy, a lower alkylthio, trifluoromethyl or trifluoromethoxy; R<3> is an (esterified) carboxy; R<4> , R<5> , R<6> and R<7> are each H, a halogen atom, a lower alkyl, a lower alkoxy or trifluoromethyl; and X is O or S) or its pharmaceutically permissible salt.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a glycated protein denaturing substance (Advanced Glycati) containing a 1,4-benzothiazine-2-acetic acid derivative or a pharmaceutically acceptable salt thereof as an active ingredient.
on End Products, hereinafter referred to as "AGE") production inhibitor.

[0002]

BACKGROUND OF THE INVENTION Non-enzymatic glycation of proteins in vivo leads to direct modification of protein function and damage of cells through receptors on cell membranes, resulting in various diseases associated with diabetes and arteriosclerosis. It is known to cause various physiological disorders. That is, glucose in blood non-enzymatically forms a Schiff base with an amino group of a protein, and further produces a rearrangement product by Amadori rearrangement. This Amadori rearrangement product releases active oxygen and is converted into an active intermediate like 3-deoxyglucosone by the action of a trace amount of a transition metal, and further reacts with an amino group of a protein, followed by oxidation, condensation and rearrangement. AGE through complicated reactions such as
Is formed. AGE is considered to be a polymer in which a protein (or a degradation product thereof) is crosslinked with a structure derived from sugar. Glycated proteins are also produced in healthy people and increase with age, but in diabetics, high glucose concentration in blood causes rapid glycation of proteins, which has a direct effect on protein function and AGE. Receptor-mediated cellular responses lead to various disorders, which are considered to be one of the causes of various diabetic complications. In addition, active oxygen generated when glycated proteins are produced is also considered to cause cell damage.

More specifically, it has been recognized that diabetic cataract is caused by polymerization, insolubilization, fluorescence generation and coloring of crystallin accompanying glycation of crystallin in the eye lens (J. Biol. Chem., 256, 5176, 1981), and diabetic neuropathy is thought to be caused by non-enzymatic glycation of neuronal myelin protein (Clin. Endocrinol. Met.
ab., 11 , 431, 1982). It is also considered that the process of AGE production is a cause of aging. For example, senile cataract is involved in glycation of crystallin in the eye lens, and AGE production is also involved in atherosclerosis. . Furthermore, it is known that AGE is also involved in the thickening of the basement membrane of thin blood vessels associated with aging and the thickening of the renal glomerular basement membrane, which causes a decrease in renal function (Science, 232, 1629,
1986).

[0004]

Under such a background, a substance that inhibits AGE formation of proteins has been searched for, and aminoguanidine has been reported as such a compound (Science, 232). , 1629, 1986). Aminoguanidine blocks the active carbonyl group of the Amadori rearrangement product and inhibits the cross-linking polymerization of the Amadori rearrangement product, and is therefore considered to inhibit the transfer of the protein to AGE. However, the action of aminoguanidine cannot be said to be sufficient, and AGE that can achieve a practically satisfactory effect is obtained.
No production inhibitor has been found yet. The present inventors
Various diseases caused by AGE production by inhibiting GE production, such as neuropathy, retinopathy, nephropathy, cataract, coronary heart disease, peripheral circulatory disorder, cerebrovascular disorder, arteriosclerosis , A compound useful for preventing and treating arteriosclerosis, cancer, Alzheimer's disease, etc., and as a result, a 1,4-benzothiazine-2-acetic acid derivative represented by the following general formula (I) or its pharmaceutically It was found that acceptable salts have excellent AGE production inhibitory activity.
The present invention has been made based on such findings.

The present invention aims to provide an AGE production inhibitor. Another object of the present invention is to provide a prophylactic / therapeutic agent for various diseases as described above which develop due to the production of AGE.

[0006]

The present invention has the following general formula (I):

[0007]

[Chemical 3]

(In the formula, R ¹ and R ² may be the same or different and each is a hydrogen atom, a halogen atom, a lower alkyl group, a lower alkoxy group, a lower alkylthio group,
Represents a trifluoromethyl group or a trifluoromethoxy group, R ³ represents an optionally esterified carboxy group, R ⁴ , R ⁵ , R ⁶ and R ⁷ may be the same or different and each is a hydrogen atom, It represents a halogen atom, a lower alkyl group, a lower alkoxy group or a trifluoromethyl group, and X represents an oxygen atom or a sulfur atom. ), A 1,4-benzothiazine-2-acetic acid derivative or a pharmaceutically acceptable salt thereof as an active ingredient.

The present invention also occurs due to the formation of AGE containing a 1,4-benzothiazine-2-acetic acid derivative represented by the above general formula (I) or a pharmaceutically acceptable salt thereof as an active ingredient. The present invention relates to a preventive / therapeutic agent for diseases.

In the compound represented by the general formula (I),
The definition of each substituent is as follows. Examples of the halogen atom include a fluorine atom, a chlorine atom, a bromine atom and an iodine atom.

The lower alkyl group is preferably a linear or branched alkyl group having 1 to 6 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl,
Examples include isopentyl, hexyl and isohexyl.

The lower alkoxy group has 1 to 6 carbon atoms.
A straight chain or branched chain alkoxy group of, for example, methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, sec-butoxy, tert-
Examples thereof include butoxy, pentyloxy, isopentyloxy, hexyloxy, isohexyloxy and the like.

The lower alkylthio group has 1 to 1 carbon atoms.
6 is preferably a linear or branched alkylthio group, and examples thereof include methylthio, ethylthio, propylthio, isopropylthio, butylthio, isobutylthio, sec-butylthio, tert-butylthio, pentylthio, isopentylthio, hexylthio and isohexylthio. To be

As the esterified carboxy group,
For example, lower alkoxycarbonyl such as methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, isopropoxycarbonyl, butoxycarbonyl, isobutoxycarbonyl, tert-butoxycarbonyl; cycloalkyloxy moieties such as cyclohexyloxycarbonyl and cyclopentyloxycarbonyl having 5 to 10 carbon atoms. Or a halogen atom, a lower alkyl group, a lower alkoxy group on the benzene ring,
Examples thereof include aryloxycarbonyl or benzyloxycarbonyl which may have a substituent such as a nitro group.

The compound represented by the general formula (I) has an asymmetric carbon atom and therefore can exist as a stereoisomer, and can be resolved into a pure isomer if necessary.

The pharmaceutically acceptable salt of the compound represented by formula (I) is not particularly limited as long as it is a pharmaceutically acceptable non-toxic salt, and examples thereof include lithium salt, sodium salt and potassium salt. Alkali metal salts such as salts; alkaline earth metal salts such as calcium salts and magnesium salts; aluminum salts; salts with organic bases such as triethylamine salts and pyridine salts; salts with basic amino acids such as lysine salts and arginine salts Can be mentioned.

In the general formula (I), R ⁴ , R ⁵ and R ⁶
R ⁷ and R ⁷ may be the same or different and each is preferably a hydrogen atom, a fluorine atom or a chlorine atom as the compound of the present invention.

In the present invention, as the compound represented by the general formula (I), for example, the compounds described in JP-A-5-92961 can be used. Typical examples of the compound represented by the general formula (I) include 3,4-dihydro-3-oxo-4-[(4,5,5
7-trifluoro-2-benzothiazolyl) methyl]-
2H-1,4-benzothiazine-2-acetic acid may be mentioned.

The compound represented by the general formula (I) can be prepared, for example, by the method described in JP-A-5-92961.

The AGE production inhibitor of the present invention comprises a compound represented by the general formula (I) or a pharmaceutically acceptable salt thereof in an appropriate pharmaceutically acceptable additive (eg carrier, excipient). Agents, diluents, etc.), and then administered orally in the form of powder, granules, tablets, capsules, etc., and parenterally in the form of injections, eye drops, etc. can do. An effective amount of the compound represented by the general formula (I) or a pharmaceutically acceptable salt thereof is added to the above-mentioned preparation. The dose varies depending on the administration route, symptoms, weight of the patient, age and the like, but for example, when orally administered to an adult patient, 1 to 2000 mg / kg body weight / day, particularly 10 to 600 mg / kg body weight / day is recommended. It is desirable to administer from once to several times a day.

[0021]

INDUSTRIAL APPLICABILITY The AGE production inhibitor of the present invention exhibits an excellent AGE production inhibitory action on mammals (eg, mouse, rat, rabbit, dog, cat, human etc.) and has low toxicity. Therefore, various diseases caused by generation of AGE, such as neuropathy, retinopathy, nephropathy, cataract, coronary heart disease, peripheral circulatory disorder, cerebrovascular disorder, arteriosclerosis, arteriosclerosis, and cancer. , Prevention of Alzheimer's disease, etc.
Useful for treatment. In particular, the AGE production inhibitor of the present invention is
Since it has an excellent inhibitory effect on glycation of hemoglobin, it is useful for the above-mentioned diseases based on tissue hypoxia resulting from functional abnormality due to glycation of hemoglobin.

[0022]

EXAMPLES The present invention will be described in more detail based on the following test examples, but the present invention is not limited to these examples. Formulation Example 1 Tablet (1) Compound A 10 mg (2) Fine granules for direct compression No. 209 (manufactured by Fuji Chemical Co., Ltd.) 46.6 mg Magnesium metasilicate aluminate 20% Corn starch 30% Lactose 50% (3) Crystalline cellulose 24.0 mg (4) Carboxymethyl cellulose / calcium 4.0 mg (5) Magnesium stearate 0. 4 mg compound A: 3,4-dihydro-3-oxo-4-
[(4,5,7-Trifluoro-2-benzothiazolyl) methyl] -2H-1,4-benzothiazine-2-acetic acid (the same applies to the following formulation examples) (1), (3) and (4) are all Also pass through a 100 mesh screen beforehand. After drying (1), (3) and (4) and (2) respectively to reduce the water content to a constant value,
Mix using a mixer in the above weight proportions. (5) is added to the mixed powder which is homogenized and mixed for a short time (30 seconds), and the mixed powder is tableted (pestle: 6.3 mmφ, 6.0 mmR).
Then, one tablet of 85 mg was prepared. If necessary, the tablets may be coated with a commonly used gastric film-coating agent (for example, polyvinyl acetal diethylaminoacetate) or an edible coloring agent.

Formulation Example 2 Capsule (1) Compound A 50 g (2) Lactose 935 g (3) Magnesium stearate 15 g The above components were weighed and mixed uniformly, and 200 mg each of the mixed powder was filled into a hard gelatin capsule. did.

Formulation Example 3 Injection (1) Compound A 50 mg (2) Glucose 100 mg (3) Physiological saline 10 ml After filtering the above mixed solution with a membrane filter, sterilization filtration was performed again, and the filtrate was aseptically filtered. Dispense into vials,
It was filled with nitrogen gas to give an intravenous injection.

Pharmacological Test In order to show the usefulness of the AGE production inhibitor of the present invention, the results of the pharmacological test of representative compounds are shown below. Test Example 1 (AGE production inhibitory activity) Test compound 3,4-dihydro-3-oxo-4-[(4,5,7-
Trifluoro-2-benzothiazolyl) methyl] -2H
-1,4-Benzothiazine-2-acetic acid (Compound A)

[0026]

[Chemical 4]

Test Method Male Sprague-Dawley rat (8 weeks old)
Streptozotocin (STZ, 60 mg / kg body weight) dissolved in a citrate buffer (pH 4.5) was intravenously administered to the tail. Three to five weeks after administration of STZ, Compound A (30 m
(g / kg body weight) was suspended in 0.5% carboxymethylcellulose and orally administered once a day for 2 weeks.
The control group was orally administered with vehicle (0.5% carboxymethylcellulose) 5 ml / kg body weight. On the last day of drug administration, a blood sample was collected using a syringe containing heparinized saline, and the blood sample was collected at 2000 rpm at 4 ° C. for 10 days.
It was centrifuged for a minute and hemoglobin was extracted.

A competitive ELISA method (J. Biol. Chem., 267, 5133 (199
2)) causes blood AGE hemoglobin (Hb-AG
E) was measured. Briefly, 50 μL of the hemoglobin solution extracted above is added to an equal volume of polyclonal anti-
AGE modified ribonuclease (diluted 1: 400) with AGE antibody (10 μg / in phosphate buffered saline).
(mL) precoated microtiter plate. After incubation at room temperature for 2 hours, horseradish peroxidase-labeled secondary antibody was added, incubated again at room temperature for 1 hour, and bound anti-AGE antibody was detected by adding o-phenylenediamine. AGE immunoreactivity was measured in duplicate on serially diluted samples. The AGE unit was calculated as a relative value to a synthetic AGE-ribonuclease standard (1 AGE unit = AGE-ribonuclease 10 μg). All data are mean ± S.E.M. E. Expressed as Dunnett's method was used for the test of significant difference.

Results The results are shown in Table 1 and FIG. 1 (#p <0.05, for the normal group; ** p <0.01, for the vehicle-administered control group).

[0030]

[Table 1]

The Hb-AGE level of the control group was the same as that of the normal group.
Significantly higher compared to b-AGE levels (Δ% = 4
5, p <0.05, relative to the normal group). In contrast, administration of Compound A significantly reduced the Hb-AGE level (p <0.01, Δ% = 48 with respect to the vehicle-administered control group). As shown in Table 1 and FIG. 1, the test compound of the present invention exhibited a high AGE production inhibitory activity.

[Brief description of drawings]

FIG. 1 is a view showing the AGE production inhibitory effect of the inhibitor of the present invention.

─────────────────────────────────────────────────── ─── Continuation of front page (51) Int.Cl. ⁷ Identification code FI theme code (reference) A61K 31/54 ABX A61K 31/54 ABX ACV ACV ADU ADU // C07D 417/06 277 C07D 417/06 277 ( 72) Inventor Tomoyuki Maruyama 2-25-1 Otani Otani, Hirakata City, Osaka Prefecture Midori Cross Research Institute Ltd. (72) Toshiaki Akira 2-25-1 Otani Otani, Hirakata City, Osaka Prefecture Incorporated Company Midori Cross Central Research Laboratory (72) Inventor Takashi Awata 1-2-1-22 Tsukaguchi-cho Amagasaki-shi Hyogo Prefecture F-term (reference) 4C063 AA01 BB03 CC64 DD62 EE01 4C086 AA01 AA02 BC89 GA10 MA01 MA04 NA14 ZA02 ZA16 ZA33 ZA36 ZA40 ZA40 ZA40 ZA40 ZA40 ZA40 ZA45 ZB26 ZC21

Claims (6)

[Claims] 1. A compound represented by the general formula (I): (In the formula, R ¹ and R ² may be the same or different and each represents a hydrogen atom, a halogen atom, a lower alkyl group, a lower alkoxy group, a lower alkylthio group, a trifluoromethyl group or a trifluoromethoxy group,
³ represents an optionally esterified carboxy group,
R ⁴ , R ⁵ , R ⁶ and R ⁷ may be the same or different and each represents a hydrogen atom, a halogen atom, a lower alkyl group, a lower alkoxy group or a trifluoromethyl group, and X represents an oxygen atom or a sulfur atom. . ) 1,4-benzothiazine-2-acetic acid derivative or a pharmaceutically acceptable salt thereof as an active ingredient, a glycated protein denaturing substance formation inhibitor. 2. The active ingredient is 3,4-dihydro-3-oxo-4-[(4,5,7-trifluoro-2-benzothiazolyl) methyl] -2H-1,4-benzothiazine-2-acetic acid. The glycated protein denatured substance production inhibitor according to claim 1, which is a pharmaceutically acceptable salt thereof. 3. A compound represented by the general formula (I): (In the formula, R ¹ and R ² may be the same or different and each represents a hydrogen atom, a halogen atom, a lower alkyl group, a lower alkoxy group, a lower alkylthio group, a trifluoromethyl group or a trifluoromethoxy group,
³ represents an optionally esterified carboxy group,
R ⁴ , R ⁵ , R ⁶ and R ⁷ may be the same or different and each represents a hydrogen atom, a halogen atom, a lower alkyl group, a lower alkoxy group or a trifluoromethyl group, and X represents an oxygen atom or a sulfur atom. . ) A preventive / therapeutic agent for a disease caused by the production of a glycated protein-denaturing substance containing a 1,4-benzothiazine-2-acetic acid derivative represented by the formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient. 4. The preventive / therapeutic agent according to claim 3, wherein the disease caused by the production of a glycated protein denatured substance is a disease caused by a functional abnormality due to glycation of hemoglobin. 5. The preventive and therapeutic agent according to claim 3 or 4, wherein the disease caused by the production of the glycated protein denatured substance is not a diabetic complication. 6. The active ingredient is 3,4-dihydro-3-oxo-4-[(4,5,7-trifluoro-2-benzothiazolyl) methyl] -2H-1,4-benzothiazine-2-acetic acid. Alternatively, a prophylactic / therapeutic agent for a disease caused by the production of the glycated protein denatured substance according to any one of claims 3 to 5, which is a pharmaceutically acceptable salt thereof.