JPH0148882B2 - - Google Patents

Description

[Detailed description of the invention]

TECHNICAL FIELD The present invention relates to a prophylactic and therapeutic agent for renal diseases containing a cyclohexanecarboxylic acid amide derivative as an active ingredient. The cyclohexanecarboxylic acid amide derivative which is an active ingredient in the renal disease preventive and therapeutic agent of the present invention has the general formula (In the formula, A is a lower alkyl group having 1 to 4 carbon atoms, a lower alkoxy group having 1 to 4 carbon atoms, chlorine,
Halogen atoms such as bromine, fluorine, and iodine, nitro groups,
Phenyl group and pyridyl group substituted with 1 to 3 hydroxyl group, carboxyl group or lower alkoxycarbonyl group at any position, R is an alkyl group, R ¹
is a hydrogen atom or a methyl group). To explain R and A in the general formula () more specifically, the alkyl group of R is methyl, ethyl, n-propyl, isopropyl, n-
Represents a lower alkyl group such as butyl, isobutyl, t-butyl, n-pentyl, isopentyl, n-hexyl, isohexyl, n-heptane, isoheptane, n-octane, isooctane, etc.
Further, A represents a substituted phenyl group or a substituted pyridyl group, and the substituent is, for example, methyl, ethyl, n-
Lower alkyl groups such as propyl, isopropyl, n-butyl and isobutyl groups, lower alkoxy groups such as methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy and isobutoxy groups,
These include halogen atoms such as chlorine, bromine, fluorine, and iodine, nitro groups, hydroxyl groups, carboxyl groups, and lower alkoxycarbonyl groups. Now, some of the compounds represented by the general formula () are already known in Japanese Patent Application Laid-open No. 55-20738, and their use is only known as an anti-windows agent having a remarkable anti-allergic effect. However, the present inventors conducted a more detailed study on the pharmacological action of the compound represented by the general formula (), and found that the compound has a completely different anti-acute action, and has excellent anti-complement action and platelet aggregation action. It has been found that it has suppressive effects, anti-inflammatory effects, cellular immunosuppressive effects, etc., and has extremely low toxicity.
That is, the present invention has found that the above-mentioned pharmacological action of the compound represented by the general formula () is useful for preventing and treating renal diseases including immunological and non-immunological nephritis. This is the completed version. Furthermore, the present invention relates to a prophylactic and therapeutic agent for renal diseases characterized by containing as an active ingredient a compound represented by the general formula () for which no medicinal use is described or suggested in known literature. Currently, there is no radical treatment for nephritis, and general symptomatic treatment is used.Currently, drug therapy includes α-methyldopa and β-blockade when hypertension is present. Diuretics such as furosemide are used when the swelling is severe, and anticholesterol drugs are also used because hypercholesterolemia occurs when nephrosis occurs. When nephritis occurs as a result of an antigen-antibody reaction, antibiotics are used to treat antigens such as β-hemolytic streptococci, and immunosuppressants such as steroids and cyclophosphamide are used to suppress antibody production. Steroids, non-steroidal anti-inflammatory drugs such as indomethacin and flufenamic acid are used to treat inflammation associated with antigen-antibody reactions and other nephritis-causing factors, as well as membranoproliferative glomerulonephritis and active lupus nephritis. For blood clotting that is said to be more likely to occur when there is active inflammation due to rapidly progressing nephritis, such as late preeclampsia and hemolytic uremic syndrome, anticoagulant therapy with heparin, warflin, etc., dipyridamole, indomethacin,
and platelet aggregation inhibitors such as aspirin, and
Urokinase and the like are also used for the purpose of dissolving preformed fibrin. Next, a method for producing the active ingredient in the present invention will be described. The compound represented by the above general formula () can be produced by the method described in JP-A No. 55-20738 or a method analogous thereto. For example, as shown in the reaction formula below, by reacting a compound represented by general formula () with a reactive derivative of a cyclohexanecarboxylic acid derivative represented by general formula (), the target compound, a compound represented by general formula (), can be obtained. can be obtained. To explain the above production method in more detail, a reactive derivative (for example, an acid halide, an ester derivative, an acid anhydride) of a cyclohexanecarboxylic acid derivative represented by the general formula () is added to the amine represented by the general formula () by tetrahydrofuran, The reaction may be carried out in an organic solvent such as acetone, chloroform, pyridine, benzene, or toluene. Further, when using an acid halide, the reaction proceeds quickly by using a deoxidizing agent (for example, trimethylamine, triethylamine, N-dimethylaniline, pyridine, sodium carbonate, potassium carbonate, etc.). or,
Among the cyclohexanecarboxylic acid amide derivatives obtained by this method, the ester form can be further hydrolyzed as desired. Hydrolysis involves water, acetic acid,
Methanol, ethanol and water and alcohols,
The reaction may be carried out in a mixed solvent such as water and acetic acid in the presence of an acid (eg, hydrochloric acid, sulfuric acid, etc.) or an alkali (eg, potassium hydroxide, sodium hydroxide, etc.) at room temperature or under heating. The reaction conditions are appropriately selected depending on the solvent used, the amount of acid or alkali, and the temperature. If desired, non-toxic salts conventionally known in the art can be used. Specifically, for example, alkali metal salts such as sodium and potassium;
Examples include salts of alkaline earth metals such as calcium, magnesium, and aluminum, and salts of various basic amino acids, such as lysine, arginine, ornithine, and the like. Table 1 specifically shows the compounds represented by the general formula ().

【table】

【table】

【table】

[Table] Next, the pharmacological action and toxicological properties of this compound will be described. First, experimental results regarding the anti-complement effects of various compounds in vitro and in vivo will be shown. Experimental example 1 Experimental method Mayer MMet al (Immunochemistry 7 ,
485, 1970). That is,
Guinea pig fresh pool serum ( _CH50 =
Complement was reacted with gelatin veronal buffer containing previously prepared sensitized sheep red blood cells (5 x 10 ⁸ ) at 37°C for 60 minutes, and the degree of hemolysis was measured using a spectrophotometer at 541 nm. Absorbance was measured. All test compounds were dissolved in physiological saline containing 10% dimethyl sulfoxide (hereinafter referred to as DMSO), and diluted with buffer as appropriate so that the final concentration of DMSO was 1%. The results are expressed as % inhibition with respect to OD ₅₄₁ when only the solvent is added as a control. The results are shown in Table 2.

[Table] Experimental Example 2 Experimental Method Male Hartley guinea pigs weighing 300 to 400 g were used in groups of 5, and Glovsky M. et al (J.
Immunol., 102 , 1, 1969). That is, inactivated anti-sheep red blood cell rabbit serum (Forssmann antibody, hemolysin titer 4096 times) 0.5
ml/animal was administered intravenously to induce shock.
The test compound was orally administered 1 hour before serum transfer. The results were expressed as survival rate and survival time. The results are shown in Table 3.

[Table] As is clear from the results in Tables 2 and 3, this compound has an excellent anti-complement effect, and furthermore, it has an excellent effect on prolonging life in response to in vivo formal shock. prevention of mortality and mortality was observed. Next, the anti-inflammatory effect of this compound will be demonstrated in carrageenan paw edema. Experimental Example 3 Experimental Method Tests were conducted using the paw volume method using male Wistar rats weighing approximately 150 g. That is, after orally adding 5 ml of water to rats that had fasted from the previous day, 1.5%
Edema was induced by subcutaneously injecting 0.1 ml of carrageenan into the sole of one hind leg, and the edema intensity was measured after 3 hours. The test compound was orally administered 1 hour before edema induction. The results were expressed as edema intensity and inhibition rate relative to the control group. The results are shown in Table 4.

[Table] As is clear from the above test results, this compound was found to have anti-inflammatory effects. Next, experimental results regarding the in vitro platelet aggregation inhibitory effect of this compound will be shown. Experimental example 4 Experimental method 1 Preparation of platelets According to a conventional method, fresh blood was collected from male Hartley guinea pigs by adding 1/10 volume of 3.8% sodium citrate, centrifuged at 800 rpm for 10 minutes, and the supernatant was collected. Platelet-rich plasma (PRP) was obtained and further centrifuged at 3000 rpm for 15 minutes, and the supernatant was obtained as platelet-poor plasma (PPP), which was stored on ice and used within 4 hours. 2 Aggregation measurement Aggregation measurement was performed using an aggregometer at 1100 rpm37
Changes in plasma absorbance associated with platelet aggregation were recorded at °C. The inhibitory effect is 1ml PRP (approximately 300,000/mm ³ )
Add 10μ of test compound dissolved in DMSO and PPP
After setting the recording range by
(final concentration 10 ⁻⁵ M) was added to cause aggregation. As a control, only DMSO was added to cause aggregation.
The inhibition rate at maximum aggregation was determined compared to the control. The results are shown in Table 5.

[Table] Next, test results regarding the cellular immunosuppressive effect of this compound are shown. Experimental example 5 Experimental method Using ddY male mice weighing around 24g,
Lagrange, PHet al (J.Exp.Med. 139 , 528,
The test was conducted according to the method of (1974). That is, sheep red blood cells (10 ⁷ /50μ) suspended in sterile physiological saline
) was administered subcutaneously to the right hind footpad for immunization.
On day 1, antigen (10 ⁸ /50μ) was inoculated into the other footpad, and the induced footpad swelling was measured at 24 hours and subtracted from the pre-induction value. The test compound was orally administered at the same time as the induction. The results are shown in Table 6.

[Table] As is clear from the above test results, this compound was found to have a suppressive effect on cell-mediated immunity. Next, the effect of this compound on experimental nephritis will be shown. Experimental Example 6 Experimental Method Male Wistar rats weighing around 150g were used. Heymann W. et al (Periadrics, 7 , 691,
Anti-rat renal rabbit serum prepared according to the method of (1951)
0.3 ml/100 g B.W. was injected intravenously. Urine protein and serum total cholesterol were measured 20 days after the induction of nephritis. The test compound was administered from the day before nephritis induction.
It was orally administered 20 times on consecutive days. The results are shown in Table 7.

[Table] As is clear from the above test results, this compound exhibited excellent urinary protein reduction and cholesterol-lowering effects, suggesting that this compound may be effective as a preventive and therapeutic agent for nephritis. Next, the acute toxicity value of this compound is shown. Experimental example 7 Experimental method ddY male mice weighing around 20g and 100g
Male Wistar rats were used before and after the experiment, and the observation period was one week. The results are shown in Table 8.

[Table] The acute toxicity values for the two compounds were shown in mice and rats, and the toxicity was extremely weak in both cases. Therefore, this compound has a strong anti-complementary effect in vitro and in vivo, and also has platelet aggregation inhibitory, anti-inflammatory and cellular immunosuppressive effects, and has also been shown in experiments. Excellent preventive and therapeutic effects on the onset of nephritis were observed. Based on these facts, this compound is effective against various renal diseases, including glomerulonephritis, nephrotic syndrome, nephrosclerosis, nephritis nephritis, or nephritis that accompanies other collagen diseases. It is also highly safe for long-term continuous use. Therefore, it can be said that this compound is an extremely effective preventive and therapeutic agent for nephritis when used alone or in combination with steroids or immunosuppressants for renal diseases requiring long-term treatment. Next, formulation of this substance for use as a preventive and therapeutic agent for renal diseases will be explained. The compound represented by the general formula () can be administered to animals and humans as it is or together with a conventional pharmaceutical carrier. The dosage unit form is not particularly limited and may be appropriately selected and used as required.
Examples of such dosage unit forms include oral preparations such as tablets, capsules, granules, and various oral liquid preparations, and parenteral preparations such as injections and suppositories. The amount of the active ingredient to be administered is not particularly limited and can be appropriately selected from a wide range, but in order to achieve the desired effect, it is preferably 0.5 to 200 mg per kg of body weight per day. In addition, the active ingredient may be contained in a dosage unit form of 25 to
It is better to contain 2000 mg. In the present invention, oral preparations such as tablets, capsules, and oral liquid preparations are manufactured according to conventional methods. That is, tablets are prepared by mixing the compound of the present invention with pharmaceutical excipients such as gelatin, starch, lactose, magnesium stearate, talcum, and gum acacia. Capsules are prepared by mixing the compound of the present invention with an inert pharmaceutical filler or diluent,
Fill into hard gelatin capsules, soft capsules, etc. Oral liquid syrups and elixirs are produced by mixing the compound of the present invention with sweeteners such as sucrose, preservatives such as methyl and propylparabens, colorants, seasonings, and the like. In addition, parenteral preparations are manufactured according to conventional methods, for example, by dissolving the compound of the present invention in a sterilized liquid carrier. The preferred carrier is water or saline. Solutions having the desired clarity, stability and suitability for parenteral use are prepared by dissolving 25 to 2000 mg of the active ingredient in water and organic solvents and then in polyethylene glycol having a molecular weight of 200 to 5000. Preferably, such a liquid agent contains a lubricant such as sodium carboxymethyl cellulose, methyl cellulose, polyvinylpyrrolidone, or polyvinyl alcohol. Furthermore, the liquid preparation may contain bactericidal agents and fungicides such as benzyl alcohol, phenol, and thimerosal, as well as isotonic agents such as sucrose and sodium chloride, local anesthetics, stabilizers, buffers, etc., as necessary. You can leave it there. In addition, from the viewpoint of stability of drugs for parenteral administration, it is also possible to fill capsules, etc., freeze them, remove water using normal freeze-drying techniques, and reconstitute liquid preparations from the freeze-dried powder immediately before use. . As mentioned above, the use of the compound represented by the general formula ( It has remarkable pharmacological effects and safety, and is an industrially very useful drug as a preventive and therapeutic agent for renal diseases. Next, the present invention will be explained in more detail with reference to production examples and formulation examples of the compound represented by the general formula (). Production example 1 4.2g of trans-4-n-hexylcarboxylic acid,
A mixture of 5 ml of thionyl chloride and 30 ml of benzene was refluxed for 3 hours, and then the solvent and excess thionyl chloride were distilled off under reduced pressure. It was added dropwise into the solution while cooling on ice. Thereafter, the mixture was allowed to react at room temperature for 7 hours. After the reaction is complete, the solvent is distilled off under reduced pressure, water and dilute hydrochloric acid are added to the residue to make it weakly acidic, and the precipitated crystals are collected and recrystallized from ethyl acetate to form new N in the form of colorless needle-like crystals.
-(2-carboxyphenyl)trans-4-n
-Hexylcyclohexanecarboxylic acid amide 5.9g
I got it. The melting point, infrared absorption spectrum, mass spectrum, and elemental analysis values of this substance were as follows. Melting point 181-183℃ Infrared absorption spectrum
υc=o(KBr) 1690, 1674 cm ^-1 Mass spectrum M ⁺ 331 Elemental analysis value C ₂₀ H ₂₉ NO ₃ Theoretical value C: 8.82 H: 72.47 N: 4.23 Actual value C: 8.86 H: 72.44 N: 4.42 Note that the above All compounds other than those shown in the above production examples shown in Table 1 are also produced according to the above method. Formulation example 1 Preparation of tablets Mixture amount (g) N-(2-carboxyphenyl)trans-4
-n-hexylcyclohexanecarboxylic acid amide 100 Amount (g) Lactose (Japanese Pharmacopoeia) 1000 Cornstarch (Japanese Pharmacopoeia) 500 Crystalline cellulose (Japanese Pharmacopoeia) 500 Methyl cellulose (Japanese Pharmacopoeia) 30 Stearin Magnesium acid salt (Japanese Pharmacopoeia product) 20 Mix the above active ingredient compound of the present invention, lactose, cornstarch and crystalline cellulose thoroughly,
The mixture was carefully dried through a mesh sieve, and then compressed into tablets containing 100 mg of the active ingredient using a conventional method. Formulation example 2 Preparation of capsules Compounding amount (g) N-(2-carboxyphenyl)trans-4
-n-hexylcyclohexanecarboxylic acid amide 100 Amount (g) Lactose (Japanese Pharmacopoeia) 800 Starch (Japanese Pharmacopoeia) 300 Talc (Japanese Pharmacopoeia) 50 Magnesium stearate (Japanese Pharmacopoeia) 20 Above Each component was finely powdered, sufficiently stirred to form a homogeneous mixture, then filled into gelatin capsules for oral administration having desired dimensions, and tableted to uniformly contain 100 mg of the active ingredient.

Claims (1)

[Claims] 1. General formula (In the formula, A is a lower alkyl group having 1 to 4 carbon atoms, a lower alkoxy group having 1 to 4 carbon atoms, chlorine,
Halogen atoms such as bromine, fluorine, and iodine, nitro groups,
Cyclohexanecarboxylic acid amide represented by a phenyl group or pyridyl group optionally substituted with 1 to 3 hydroxyl groups, carboxyl groups, or lower alkoxycarbonyl groups, R is an alkyl group, and R ¹ is a hydrogen atom or a methyl group) A novel renal disease prevention and treatment agent containing a derivative as an active ingredient.