JPH09268125A - Therapeutic medicine for nephritis - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain the subject preparation containing a hydroxamic acid derivative, having a mesangium cell proliferation-inhibiting action and capable of being effectively used as a therapeutic medicine for nephritis. SOLUTION: This preparation contains a hydroxamic acid derivative of the formula [R<1> is an aryl, etc.,; L is an alkylene, etc., (n) is an integer of 0 or 1; R<2> is H, an alkyl, etc.,; M is H, a medically acceptable cation, etc.], e.g. [N-hydroxy-N-(1-phenylethyl)-4-(3-(4-methoxyphenoxy) styryl)benzamide]. The hydroxamic acid derivative can be synthesized by a method described in a Japanese unexamined patent: Hei 7-278086, etc., and can remarkably depress the proliferation of cultured rat masangium cells.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a pharmaceutical preparation containing a hydroxamic acid derivative which has a mesangial cell growth inhibitory action and is effective as a therapeutic drug for nephritis.

[0002]

2. Description of the Related Art Proliferative glomerulonephritis is a primary glomerulonephritis whose main lesions are proliferation of mesangial cells and increase of mesangial matrix. In particular, proliferation of mesangial cells and increase of substrate are common findings in many glomerular diseases such as IgA nephropathy, membranous proliferative nephritis, focal glomerulosclerosis, and lupus nephritis. It is assumed that various cytokine / growth factor production enhancement, active oxygen production, and the like are involved in the proliferation of mesangial cells and the increase of substrates. As a treatment for these glomerulonephritis, various drug treatments such as antiplatelet drugs, steroids, and anticoagulants are performed. However, it has not been possible to subside the proliferative lesions of the glomerulus and prevent the progression of the glomerular loop to irreversible damage, and an effective treatment method for progressive proliferative glomerulonephritis has been established. The current situation is that there are none. Therefore, the emergence of active substances that more strongly suppress the proliferation of mesangial cells is strongly desired.

[0003]

Therefore, an object of the present invention is to provide a therapeutic agent for nephritis containing a compound useful as a therapeutic agent for nephritis as an active ingredient.

[0004]

Means for Solving the Problems The present inventors disclosed in Japanese Patent Laid-Open No.
Regarding the hydroxamic acid derivative described in JP-A-278086, as a result of diligent examination of their pharmacological activities, it was found that the proliferative effect of serum-induced cultured mesangial cells was specifically suppressed, and the following inventions were completed. . The present invention is as follows. The present invention is a pharmaceutical preparation containing a hydroxamic acid derivative represented by the following general formula (1).

[0005]

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In the formula 1, R ¹ represents an aryl group or an aryloxyphenyl group or the following general formula (2), and L represents alkylene, alkenylene or-(CH ₂ ) m.
—O— (m is 0 or an integer of 1 to 4) and —CO—, n is an integer of 0 or 1, R ² is hydrogen, an alkyl group or an arylalkyl group, and M is hydrogen. , A pharmaceutically acceptable cation, and a group capable of being metabolically cleaved.

[0007]

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In formula 2, R ³ represents an aryl group or an arylalkyl group.

In the present invention, "alkylene" is a divalent group derived from a linear or branched alkane, such as --CH _2- , --CHCH _3- , --C (CH ₃ ) _2- ,
_{-CH (C 2 H 5) -} , - CH 2 CH 2 -, - CH 2 CHC
_{H 3 -, - C (CH} 3) 2 C (CH 3) 2 -, - CH 2 CH 2
CH ₂ — and the like. The term "alkenylene", divalent group derived from a straight-chain or branched alkenes, for example, -CH = CH -, - CH = CHCH 2 -, - CH =
_{CHCH (CH 3) -, -} C (CH 3) = CHCH 2 -,
_{-CH 2 CH = CHCH 2 -,} - C (CH 3) 2 CH = CH
C (CH ₃₎ shows 2 and the like.

The term "alkyl" refers to straight or branched chain groups and cyclic groups, including methyl group, ethyl group, propyl group, isopropyl group, butyl group, sec-butyl group, isobutyl group, tert-group. Butyl group, pentyl group,
Examples thereof include, but are not limited to, an isopentyl group, a hexyl group, a heptyl group, an octyl group, a nonyl group and a decyl group. "Alkoxy" means -OR ⁴
(R ⁴ represents an alkyl group), which includes a methoxy group, an ethoxy group, a propoxy group, an isopropoxy group,
Butoxy group, sec-butoxy group, isobutoxy group, t
The ert-butoxy group and the like are included, but the present invention is not limited thereto.

The term "aryl" means a substituted or unsubstituted carbocyclic or heterocyclic aromatic group (substituents are halogeno group, nitro group, cyano group, alkyl group, alkoxy group,
And a halogen-substituted alkyl group),
This includes phenyl groups, 1- or 2-naphthyl groups, 2
It includes, but is not limited to, a-, 3- or 4-pyridyl group, a 2- or 3-furyl group, and the like. “Aryloxy” refers to —OR ⁵ (R ⁵ represents an aryl group), and includes, but is not limited to, a phenoxy group, a 1-naphthoxy group, a 2-naphthoxy group and the like. Absent.

The term "arylalkyl" refers to an alkyl group to which an aryl group is bonded, which includes phenylmethyl group (benzyl group), 1-phenylethyl group, 2-phenylethyl group, 1-naphthylethyl group. , 2-pyridylmethyl group, benzhydryl group and the like, but are not limited thereto. “Halogen” refers to a group derived from a fluorine atom, a chlorine atom, a bromine atom, or an iodine atom.

"Halogen-substituted alkyl" refers to the above alkyl groups substituted with one or more halogens, including chloromethyl, trifluoromethyl,
Examples include, but are not limited to, 2,2-difluoroethyl groups. “Pharmaceutically acceptable cation” refers to non-toxic cations including, but not limited to, cations based on alkali and alkaline earth metals such as sodium, potassium, magnesium. .

"Metabolically cleavable group" refers to a residue to which the compound to which it is attached is easily cleaved in vivo, the compound remaining physiologically active after the cleavage. Or becomes physiologically active after cleavage. This includes
Examples include, but are not limited to, an alcoyl group, an alkoxycarbonyl group, and a peptidyl group composed of 1 to 5 amino acid residues selected from α-amino acids. The term "alcoyl" refers to -COR ⁶ (R ⁶ represents an alkyl group or hydrogen), which includes a formyl group, an acetyl group, a propionyl group, a butyryl group, an isobutyryl group, a pivaloyl group, It is not limited to these.

"Alkoxycarbonyl" means --COR.
⁷ (R ⁷ represents an alkoxy group), including methoxycarbonyl group, ethoxycarbonyl group, isopropoxycarbonyl group, butoxycarbonyl group, sec-butoxycarbonyl group, isobutoxycarbonyl group, te
Examples thereof include, but are not limited to, an rt-butoxycarbonyl group. "Peptidyl group" means C
-Shows a single amino acid linked to the parent compound at the -terminus, or two or more amino acids linked by a peptide bond bound to the parent compound at the C-terminus. These include, but are not limited to, glycyl, alanyl, valyl, lysyl, alanylalanyl, and the like.

[0016]

BEST MODE FOR CARRYING OUT THE INVENTION The hydroxamic acid derivative of the present invention can be administered orally or parenterally (eg, intravenously, intramuscularly, subcutaneously) as a therapeutic drug for nephritis. The dose of the compound of the present invention will vary depending on the age, weight and symptoms of the patient, but is usually about 0.1 to 1000 mg / kg per day,
Preferably, 1-100 mg / kg is administered in 1 to 3 divided doses.

The compound of the present invention is used as an active ingredient or one of the active ingredients, alone or in combination with a pharmaceutical carrier, according to known formulation techniques, such as tablets, powders, capsules, granules, syrups, solutions, suspensions, injections, Any formulation suitable for administration such as eye drops, liposomes, emulsions, lipid microspheres or suppositories can be used. Specific formulation carriers include starch, sucrose, lactose, methyl cellulose, carboxymethyl cellulose, crystalline cellulose, sodium alginate, calcium hydrogen phosphate, magnesium aluminometasilicate, anhydrous silicic acid,
And excipients such as synthetic aluminum silicate, binders such as hydroxypropylcellulose, hydroxypropylmethylcellulose, gelatin and polyvinylpyrrolidone, disintegrants such as carboxymethylcellulose calcium, crosslinked sodium carboxymethylcellulose and crosslinked polyvinylpyrrolidone, magnesium stearate and Lubricants such as talc, cellulose acetate phthalate, hydroxypropylmethyl cellulose acetate succinate, coating agents such as methacrylic acid and methyl methacrylate methyl copolymer, dissolution aids such as polyethylene glycol, sodium lauryl sulfate, lecithin, sorbitan monoole. Ate, polyoxyethylene cetyl ether, sucrose fatty acid ester, polyoxyethylene hydrogenated castor oil and guar Emulsifiers monostearate, etc., chelating agents such as EDTA, buffering agents, humectants, preservatives, it may be mentioned groups such as cocoa butter and Uitebuzoru W35.

The hydroxamic acid derivative of the present invention can be synthesized by the method described in JP-A-7-278086.

[0019]

The present invention will be described in more detail with reference to Reference Examples and Test Examples, but the present invention should not be limited to these Reference Examples and Test Examples.

Reference Example (a) Synthesis of N-benzyloxy-N- (1-phenylethyl) -4- (3- (4-methoxyphenoxy) styryl) benzamide 4- (3- (4-methoxyphenoxy) ) Styryl) benzoic acid (1.94 g, 5.6 mmol) was added to thionyl chloride (10
ml) was added and the mixture was stirred at room temperature for 1 hour. The mixed solution was concentrated under reduced pressure to obtain 4- (3- (4-methoxyphenoxy) styryl) benzoic acid chloride. Then, triethylamine (0.78 ml, 5.6 mmol) was added to a solution of O-benzyl-N- (1-phenylethyl) hydroxylamine (1.27 g, 5.6 mmol) in methylene chloride (20 ml), and the mixture was cooled with ice. Methylene chloride (8 ml) of the acid chloride obtained above
The solution was added dropwise and stirred at room temperature for 1 hour. Water was added to the reaction solution, which was extracted with methylene chloride. The methylene chloride layer was washed with water and a saturated aqueous solution of sodium chloride, dried over anhydrous magnesium sulfate, the solvent was distilled off under reduced pressure, and the resulting residue was subjected to silica gel chromatography to obtain a white solid from the fraction eluted with methylene chloride. The target compound (1.5 g, 48%) was obtained.

(B) N-hydroxy-N- (1-phenylethyl) 4- (3- (4-methoxyphenoxy))
Synthesis of styryl) benzamide N-benzyloxy-N- (1-phenylethyl) -4
-(3- (4-Methoxyphenoxy) styryl) benzamide (1.5 g, 2.7 mmol) in methylene chloride (14 m
l) To the solution under ice cooling, 1.0 M boron trichloride methylene chloride solution (3.24 ml, 3.24 mmol) was added dropwise, and the mixture was stirred at room temperature. After adding methanol to the reaction solution, the solvent was distilled off under reduced pressure, methylene chloride was added to the obtained residue, washed with water and a saturated aqueous sodium chloride solution, dried over anhydrous magnesium sulfate, and the solvent was distilled off under reduced pressure. The residue thus obtained was subjected to silica gel chromatography, and a 0.5% methanol-methylene chloride elution fraction showed the structure in the following formula (3), and the target compound (0.5) as a white solid having the following properties was obtained.
83 g, 66%) was obtained.

[0022]

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Mp: 161-163 ° C. ¹ H-NMR (60 MHz, CDCl ₃ ) δ (ppm): 1.
68 (3H, d, J = 7.0Hz), 3.81 (3H, s),
5.3 (1H, q, J = 7.0Hz), 6.7-7.6 (19
H, m) MS (FAB): 466 (M + 1)

(Test Example) Growth inhibitory effect on cultured rat mesangial cells Glomeruli were isolated from 6-week-old male SD rats (Charles River Japan, Inc.) by the sieving method. 20% fetal calf serum (FCS), 1M HEPE
90 mm tissue culture dish (FALCON) suspended in RPMI1640 medium containing S and ITS Premix
, And primary culture was performed in an incubator under the conditions of 37 ° C., 95% O ₂ + 5% CO ₂ . After about 1 month of primary culture, the cells were detached with EDTA trypsin and subcultured.
The test was performed using cells at passage number 2 to 3.

2 × 10 ^{4 of the} above mesangial cells were placed in a 24-well plate containing a cover glass (φ13 mm).
The seeds were seeded at a ratio of individual / hole. After culturing for 72 hours, serum-free culture was carried out for 48 hours. The medium for cell growth (RPMI1640 medium containing 20% FCS) was replaced with the hydroxamic acid derivative of the present invention obtained in the above Reference Example (18 hours). At that time, the upper limit of the final concentration of DMSO used as the solvent was 0.05%.

³ H-thymidine was added at 1 μCi per well and incubated for 6 hours. The cells were washed twice with PBS and then allowed to stand for 10 minutes with 5% perchloric acid. After replacing 5% perchloric acid with ethanol / ether (3: 1), the coverslips were removed and transferred to liquid scintillation vials. Liquid scintillation cocktail (Optiflow) was added and the radioactivity was measured with a liquid scintillation counter. The results are shown in Table 1.

[0027]

[Table 1]

As shown in Table 1, the hydroxamic acid derivative of the present invention markedly suppressed the growth of cultured mesangial cells.
The 50% inhibitory concentration in the table means that the proliferation ability of the cultured mesangial cells when not treated with the hydroxamic acid derivative of the present invention is 100%, and the proliferation ability of the cultured mesangial cells when treated with the hydroxamic acid derivative of the present invention. 50
It means the solution concentration of the hydroxamic acid derivative of the present invention required to be suppressed to%.

(Acute toxicity) An acute toxicity test was carried out by oral and intravenous administration using male ICR mice (5 weeks old). As a result, the LD ₅₀ of the hydroxamic acid derivative of the present invention was determined.
The values were all 1000 mg / kg or more, confirming higher safety than efficacy.

[0030]

The pharmaceutical preparation containing the hydroxamic acid derivative of the present invention can be effectively used as a drug for treating nephritis.

Continuation of front page (51) Int.Cl. ⁶ Identification number Office reference number FI Technical display area C07D 295/22 C07D 295/22 A

Claims (1)

[Claims] 1. A pharmaceutical preparation comprising a hydroxamic acid derivative represented by the following general formula (1). Embedded image In Formula 1, R ¹ represents an aryl group or an aryloxyphenyl group, or the following general formula (2), L is alkylene, alkenylene,-(CH ₂ ) m-O- (m is
0 or an integer of 1 to 4), —CO—, n is an integer of 0 or 1, R ² is hydrogen, an alkyl group or an arylalkyl group, M is hydrogen, a pharmaceutically acceptable cation. , A group capable of being metabolically cleaved is shown. Embedded image In Formula 2, R ³ represents an aryl group or an arylalkyl group.