JPH09169648A - Medicine composition - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain the subject composition excellent in inhibitory action on proliferation of mesangium cell, effective for an animal nephritis model, low in toxicity, useful for treating nephritis, containing a specific compound as an active ingredient. SOLUTION: This composition comprises a hederagenin derivative of the formula Me is CH3 ; R<1> is an OR<11> or an NR<11> R<12> [R<11> and R<12> are each H, a (substituted) alkyl, a (substituted) cycloalkyl, etc.]; R<2> and R<3> are each H, a (substituted) alkyl, a (substituted) cycloalkyl, etc.} or its pharmaceutically permissible salt or a solvate of any thereof such as hederagenin (3β,23- dihydroxyolean-12-en-28-oic acid as an active ingredient. The compound, for example, is obtained by extracting Spindus mukurossi in a raw state as it is in a hydrous alcohol solvent at 50-80 deg.C for 1-4 hours, concentrating the extracted solution under reduced pressure, hydrolyzing the concentrated solution with a solution of a mineral acid in an alcohol solvent, adjusting the solution to pH 6-7, adding activated carbon and purifying under heating.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD OF THE INVENTION

The present invention relates to a hederagenin derivative useful as a medicine, a pharmaceutically acceptable salt thereof and a solvate of any of them. The compound according to the present invention has a mesangial cell proliferation inhibitory action and is useful for the treatment of nephritis.

[0003]

BACKGROUND OF THE INVENTION Nephritis is caused by the location of the main lesion.
Classified into glomerulonephritis, interstitial nephritis, pyelonephritis, etc.
A typical one among them is glomerulonephritis in which the lesion is the glomerulus. Today, nephritis and glomerulonephritis are used interchangeably (Latest Medical Dictionary, 1st edition, 570, (198
7)). The most frequent histopathological findings in human glomerulonephritis and important for prognosis are the proliferation of mesangial cells and the proliferation of a substrate produced by mesangial cells (hereinafter referred to as mesangial substrate). The finding is Ig
It is common in most proliferative glomerulonephritis such as A nephropathy, membranous proliferative glomerulonephritis and lupus nephritis (Iida, Renal and Dialysis, 35 , 505-509, (1993)). Then, when the proliferation of mesangial cells and the production of mesangial matrix accompanying it progress, the glomeruli fall into the terminal glomerular sclerosis. Therefore, a compound that inhibits the proliferation of mesangial cells and the production of mesangial substrates is extremely useful as a therapeutic agent for glomerulonephritis.

Two processes are known for the proliferation of mesangial cells in glomerulonephritis. The first process involves complement, platelets and infiltrating cells. That is, the deposition of immune complexes on the glomerulus caused by an immunological mechanism occurs first, and then local activation of complement and platelets and infiltration of macrophages and neutrophils occur. These cells release various growth factors and cytokines, activate mesangial cells, and stimulate their growth. The second process involves the mesangial cells themselves. That is, it is a process in which activated mesangial cells themselves release various growth factors and cytokines, and the released cells themselves and adjacent mesangial cells are activated or proliferate. Thus, mesangial cells are activated and proliferate through multiple intricately entangled processes. Therefore, in consideration of the treatment of glomerulonephritis, it is possible to sufficiently suppress the proliferation of mesangial cells even if the certain stage of the above-mentioned first process, for example, the stage mediated by complement or platelet is suppressed. It is hard to think. In fact, it is said that the antiplatelet drug alone has a poor therapeutic effect on nephritis in active humans (Toi et al., Comprehensive Clinic, 38 , 865-870, (198
9)).

It is known that compounds having a therapeutic effect on nephritis exist in natural products derived from plants. For example, JP-A-61-37749 (Oleanene derivative), JP-A-61-85344 (Oleanene derivative), JP-A-2-7301
No. 2 (bryonolic acid derivative), JP-A-4-290846 (bryonolic acid derivative) and JP-A-61-43141 (lupine derivative) show that a pentacyclic triterpene derivative is useful for treating nephritis and the like. There is a statement to that effect. However, there is no disclosure of test examples showing that these compounds are useful for treating nephritis and the like, and no description of mesangial cell growth inhibitory action.

The hederagenin (3β, 23-
Dihydroxyolean-12-ene-28-oic acid) is a pentacyclic triterpene derivative, that is, a natural product obtained from mukurodi, ivy, ivy, yam and the like. Hederagenin has an antihypertensive effect, an antispasmodic effect (Ann. Pharm. Fr., 30 , 555 (1972)), promotion of hair growth, suppression of skin aging (FR 2669225), and an antifungal effect (Ann. Pharm. Fr. , 38 , 545 (1980)), anti-inflammatory effect (C
hem. Pharm. Bull., 28 , 1183 (1980)), anti-fluke action (Rastit. Resur., 28 , 103 (1992)), anti-ulcer, anti-allergic action (Proc. Asian. Symp. Med. Plants Spices,
4th, Meeting Date 1980, Volume 1, 59), sunburn prevention effect (JP-A-1-42411), body odor prevention, hyperhidrosis suppression effect (FR2541895 publication), etc. are known for the treatment of nephritis. Not known to be useful.

The hederagenin derivative is as follows:
The compounds (1) to (15) are known, but they are not known to be useful as a medicine for treating nephritis. (1) 3β-hydroxy-23 (4α) -acetoxyolean-12-ene-28-oic acid (Chem. Ph
arm. Bull., (1976), 24 (6), 1314)). (2) 3β-hydroxy-23 (4α) -methoxyolean-12-ene-28-oic acid (Chem. Phar
m. Bull., (1976), 24 (5), 1021). (3) 3β, 23 (4α) -diacetoxyolean-12
-En-28-Oic Acid (J. Chem. Soc., Che
m. Commun., (1983), (17), 939). (4) 3β, 23 (4α) -dibenzoyloxyolean-12-ene-28-oic acid (Bull. Soc. R
oy. Sci. Liege, (1973), 42 (5-6), 245). (5) 3β, 23 (4α) -diformyloxyolean-
12-En-28-Oic Acid (CAS, Registry
No. = 6055-17-0). (6) Methyl 3β, 23 (4α) -dihydroxyolean-12-en-28-oate (J. Chem. Soc., Che
M. Commun., (1981), (21), 1136). (7) Methyl 3β-hydroxy-23 (4α) -acetoxyolean-12-en-28-oate (Phytoche
mistry, (1984), 23 (3), 615).

(8) Methyl 3β-hydroxy-23 (4
α) -Methoxyolean-12-en-28-oate (Chem. Pharm. Bull., (1979), 27 (10), 2388). (9) Methyl 3β-acetoxy-23 (4α) -hydroxyolean-12-en-28-oate (Tetrahed
ron, (1993), 49 (33), 7193). (10) Methyl 3β-benzoyloxy-23 (4α)
-Hydroxyolean-12-en-28-oate (Tetrahedron, (1993), 49 (33), 7193). (11) Methyl 3β-methoxy-23 (4α) -hydroxyolean-12-en-28-oate (Chem. Ph
arm. Bull., (1982), 30 (9), 3340). (12) Methyl 3β, 23 (4α) -diacetoxyolean-12-en-28-oate (Chem. Pharm. Bul
l., (1982), 30 (9), 3340). (13) Methyl 3β-acetoxy-23 (4α) -methoxyolean-12-en-28-oate (Chem. Ph
arm. Bull., (1972), 20 (9), 1935). (14) Methyl 3β, 23 (4α) -dibenzoyloxyolean-12-en-28-oate (Bull. Soc.
Roy. Sci. Liege, (1973), 42 (5-6), 245). (15) Methyl 3β, 23 (4α) -dimethoxyolean-12-en-28-oate (Indian J. Chem., S
ect. B, (1990), 29B (5), 425).

[0009]

As described above, it is clear that the compound having the mesangial cell proliferation inhibitory action is an excellent therapeutic agent for nephritis. The inventors of the present invention have made repeated studies in search of such compounds.

[0010]

As a result, they have found that a specific hederagenin derivative has an excellent mesangial cell growth inhibitory action, and completed the present invention. The present invention is a pharmaceutical composition for treating nephritis, which comprises a hederagenin derivative represented by the following formula [1], a pharmaceutically acceptable salt thereof, or a solvate of any of them as an active ingredient.

[0011]

Embedded image

In the formula, Me represents CH ₃ . R ¹ is OR ¹¹
Alternatively, it represents NR ¹¹ R ¹² . R ¹¹ and R ¹² are the same or different and each is hydrogen, an alkyl which may have a substituent,
Cycloalkyl which may have a substituent, alkenyl which may have a substituent, alkynyl which may have a substituent, aryl which may have a substituent, or has a substituent. Represents an optionally substituted aromatic heterocyclic group. R ² and R ³ are the same or different and each represent hydrogen, alkyl optionally having substituent (s), cycloalkyl optionally having substituent (s), alkenyl optionally having substituent (s), substituent (s) Alkynyl which may have, acyl which may have a substituent, monoalkylcarbamoyl which may have a substituent, dialkylcarbamoyl which may have a substituent, or which has a substituent. Represents an optionally substituted alkoxycarbonyl, or R ²
And R ³ together represent carbonyl. R ¹¹ , R ¹² ,
The substituents in R ² and R ³ are the same or different and each is halogen, OR ¹³ , OCOR ¹³ , COOR ¹³ , cyano, NR ¹³ R ¹⁴ , cycloalkyl, halogen,
It is selected from aryl optionally substituted with alkyl, hydroxy or amino, and aromatic heterocyclic group optionally substituted with halogen, alkyl, hydroxy or amino. R ¹³ and R ¹⁴ are the same or different and each represent hydrogen, or alkyl optionally substituted with hydroxy, alkoxy, amino, monoalkylamino, or dialkylamino.

Among the hederagenin derivatives represented by the formula [1], the compound in which R ¹ is hydroxy, R ² is hydrogen and R ³ is hydrogen is a known natural product called hederagenin, It has a different chemical structure from the pentacyclic triterpene derivative described as being useful for the treatment of nephritis. For example, in olean-12-ene-3β, 22β, 23 (4β) -triol described in JP-A-61-37749, hydroxy is substituted at the 22nd position, and hydroxy at the 23rd position is β-configuration, In contrast to methyl at position 28, hederagenin has an unsubstituted structure at position 22, an α-configuration at hydroxy at position 23, and a carboxylic acid at position 28, which differ in chemical structure. Hederagenin has been known to have various physiological effects as described above, but it is not known to be useful for treating nephritis.

Among the hederagenin derivatives represented by the formula [1], the compounds except the following cases (1) to (5) are novel compounds which have not been described in the literature. (1) When R ¹ is hydroxy or methoxy, R ² is hydrogen, and R ³ is hydrogen, acetyl or methyl. (2) When R ¹ is hydroxy and R ² and R ³ are both formyl, acetyl or benzoyl. (3) When R ¹ is methoxy, R ² is acetyl, and R ³ is hydrogen, acetyl or methyl. (4) R ¹ is methoxy, R ² is benzoyl, R ³
Is hydrogen or benzoyl. (5) R ¹ is methoxy, R ² is methyl, and R ³ is hydrogen or methyl. A feature of the present invention is that the compound represented by the formula [1] has a mesangial cell proliferation inhibitory action and is useful for treating nephritis.

The terms used in the present invention are explained below.
“Alkyl” means a straight or branched chain having 1 to 7 carbon atoms, for example, methyl, ethyl, n-propyl,
Isopropyl, n-butyl, isobutyl, sec-butyl, te
rt-butyl, n-pentyl, isopentyl, n-hexyl,
Examples include isohexyl, n-heptyl, isoheptyl and the like. Particularly, straight-chain ones having 1 to 3 carbon atoms are preferable, and examples thereof include methyl, ethyl and n-propyl. "Cycloalkyl" has 3 to 7 carbon atoms.
And examples thereof include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and the like.

"Alkenyl" means a straight or branched chain having 2 to 7 carbon atoms, for example, vinyl, 1-propenyl, isopropenyl, allyl, 1-butenyl, 3
-Butenyl, 1-pentenyl, 4-pentenyl, 1-hexenyl, 5-hexenyl, 1-heptenyl, 6-heptenyl and the like can be mentioned. “Alkynyl” means a straight or branched chain having 2 to 7 carbon atoms,
For example, ethynyl, 1-propynyl, 2-propynyl,
1-butynyl, 3-butynyl, 1-pentynyl, 4-pentynyl, 1-hexynyl, 5-hexynyl, 1-heptynyl, 6-heptynyl and the like can be mentioned.

The term "aryl" means those having 6 to 10 carbon atoms, and examples thereof include phenyl, 1-naphthyl, 2-naphthyl and the like. The “aromatic heterocyclic group” means a 5- or 6-membered aromatic ring group having 1 to 4 heteroatoms selected from nitrogen, oxygen and sulfur, and includes, for example, 1-pyrrolyl, 2- Pyrrolyl, 3-pyrrolyl, 2-
Furanyl, 3-furanyl, 2-thienyl, 3-thienyl, 2-oxazolyl, 2-thiazolyl, 1H-1,
2,4-triazol-1-yl, 1H-tetrazol-5-yl, 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidinyl, 4-pyrimidinyl, 2-pyrazinyl, 1,3,5-triazine- 2-yl and the like can be mentioned.

"Halogen" means fluorine, chlorine, bromine,
And iodine. “Acyl” means “alkylcarbonyl” and “arylcarbonyl”. The alkyl part of "alkylcarbonyl" represents the above alkyl. The aryl part of "arylcarbonyl" represents the above aryl. Specific examples of the "acyl" include acetyl, propionyl, butyryl, isobutyryl, benzoyl, 1-naphthoyl, 2-naphthoyl and the like. The alkyl moieties of "alkoxy", "alkoxycarbonyl", "monoalkylcarbamoyl" and "dialkylcarbamoyl" are the same or different and represent the above alkyl.

Among the compounds [1] according to the present invention, preferred compounds as a therapeutic agent for nephritis are the following compound groups (1) to (5). (1) R ¹ is hydroxy, acyloxyalkoxy, phenylalkoxy, alkoxyalkoxy, hydroxyalkoxy, haloalkoxy, di (hydroxyalkyl) aminoalkoxy, (alkoxyalkyl) amino, halogen-substituted phenylamino, (dialkylaminoalkyl) amino, [ Di (hydroxyalkyl) aminoalkyl] amino, (carboxyalkyl) amino, (alkoxycarbonylalkyl) amino, (cyanoalkyl)
A compound which is amino or diamino substituted triazinylalkylamino, wherein R ² is hydrogen and R ³ is hydrogen. (2) R ¹ is hydroxy, (alkoxyalkyl) amino,
Halogen-substituted phenylamino, (dialkylaminoalkyl) amino, [di (hydroxyalkyl) aminoalkyl] amino, (alkoxycarbonylalkyl) amino or (cyanoalkyl) amino, R ² is acetoxy and R ³ is acetoxy. A compound. (3) A compound in which R ¹ is phenylalkoxy, R ² is hydroxy, (alkoxycarbonylalkyl) carbamoyl or (alkoxyalkyl) carbamoyl, and R ³ is phenylalkyl. (4) A compound in which R ¹ is hydroxy, R ² is (alkoxycarbonylalkyl) carbamoyl or (alkoxyalkyl) carbamoyl, and R ³ is hydrogen. (5) A compound in which R ¹ is hydroxy or acyloxyalkoxy and R ² and R ³ together are carbonyl.

Among the compounds [1] according to the present invention, more preferable compounds as therapeutic agents for nephritis are those in which R ¹ is hydroxy,
(Alkoxyalkyl) amino or acyloxyalkoxy, R ² is hydrogen, and R ³ is hydrogen. Among the compounds [1] according to the present invention, a particularly preferable compound as a therapeutic agent for nephritis is R ¹ is hydroxy, 2-methoxyethylamino or 2-acetoxyethoxy, R ² is hydrogen and R ³ is hydrogen. It is a compound.

Hederagenin can be obtained, for example, by the method shown below. Mukoji ( Sapindus mukorossi
Gaertn.) Raw or dried or dried and pulverized, and this is heated and extracted at 50 to 80 ° C. for 1 to 4 hours with water, alcohol (for example, methanol, ethanol, isopropyl alcohol, etc.) or hydrous alcohol as a solvent, and then The extract is concentrated under reduced pressure at 60 ° C or lower to obtain an extract. A 1-5% mineral acid (eg, hydrochloric acid, sulfuric acid, nitric acid, etc.) alcohol (methanol, ethanol, isopropyl alcohol, etc.) solution is added to the obtained extract, and the mixture is heated under reflux for 1 to 3 hours for hydrolysis. After hydrolysis, alkali (eg,
PH 6 to 7 with potassium hydroxide, sodium hydroxide, etc.
And Activated carbon may be added to this alcohol solution and heated for purification. Concentrate the alcohol solution under reduced pressure,
Hederagenin can be obtained. The obtained hederagenin can be purified by means of washing with a suitable solvent such as acetonitrile or ethanol, recrystallization, column chromatography, thin layer chromatography and the like.

Hederagenin has three functional groups, namely carboxy and two hydroxy. A desired hederagenin derivative [1] can be produced by using hederagenin as a starting material and utilizing the difference in reactivity of these functional groups. Hederagenin (ie R ¹ = OH, R ²
= H, R ³ = H) and the 28-position derivative (R ¹ ≠ OH, R ² =
H, R ³ = H) can be produced. 2 from the 28-position derivative using the following "2.23 position hydroxy reaction"
3- and 28-position derivatives (R ¹ ≠ OH, R ² = H, R ³ ≠ H)
Can be manufactured. From the 23- and 28-position derivatives, use the following "3.3 β-position hydroxy reaction" to obtain the 3β-position,
23- and 28-position derivatives (R ¹ ≠ OH, R ² ≠ H, R ³ ≠
H) can be produced. After producing the 23-position and 28-position derivatives from the 28-position derivative having a protecting group introduced at the 28-position using the following "reaction of 2.23-hydroxy",
The 28-position is deprotected to give the 23-position derivative (R ¹ = OH, R ² =
H, R ³ ≠ H) can be produced. 23rd and 28th
From the 23- and 28-position derivatives in which a protecting group is introduced at the 3-position, the 3β-position, 2-
After producing the 3- and 28-position derivatives, the 23- and 28-positions are deprotected to give 3β-position derivatives (R ¹ = OH, R ² ≠ H, R ³ =
H) can be produced.

23-position in which a protecting group is introduced only in 28-position,
After producing the 3β-position, 23-position, and 28-position derivative from the 28-position derivative using the following "3.3 β-hydroxy reaction", the 28-position is deprotected to give the 3β-position, 23-position derivative (R ¹
= OH, R ² ≠ H, R ³ ≠ H) can be produced.
From the 23-position and 28-position derivatives in which a protecting group is introduced only at the 23-position, use the following "3.3 β-position hydroxy reaction"
After the β-position, the 23-position and the 28-position derivative are produced, the 23-position is deprotected to give the 3β-position, the 28-position derivative (R ¹ ≠ OH, R ² ≠
H, R ³ = H) can be produced. From hederagenin (R ¹ = OH, R ² = H, R ³ = H), the following “4.3
β-position, 23-position dihydroxy reaction ”,
A 3-position derivative (R ¹ = OH, R ² ≠ H, R ³ ≠ H) can be produced. 28-position derivative (R ¹ ≠ OH, R ² = H,
From R ³ = H), the 3β position, 23 position and 28 position derivative (R
¹ ≠ OH, R ² ≠ H, R ³ ≠ H) can be produced.

1. Reaction of carboxy (1)

Embedded image [In the formula, R ² and R ³ are the same as above. R ¹¹⁰ represents R ¹¹ excluding hydrogen, X is a halogen such as chlorine, bromine or iodine,
It represents a leaving group such as alkylsulfoxy such as methanesulfoxy, arylsulfoxy such as toluenesulfoxy and the like. ]

The ester [5] can be produced by reacting the carboxylic acid [3] with the compound [4]. This reaction is usually carried out without a solvent or with an aprotic solvent (for example, acetonitrile, N, N-dimethylformamide (D
Polar solvent such as MF), tetrahydrofuran (TH
F), an ether solvent such as diethyl ether, a halogenated hydrocarbon solvent such as chloroform and methylene chloride, a hydrocarbon solvent such as benzene, toluene and n-hexane, or a mixed solvent thereof, and a base (for example, It can be carried out at -20 to 100 ° C in the presence of potassium carbonate, sodium carbonate, sodium hydrogen carbonate, potassium hydrogen carbonate, pyridine, 4-dimethylaminopyridine, triethylamine, sodium hydride and the like. The reaction time varies depending on the kinds of the compound [3] and the compound [4] and the reaction temperature, but is usually suitable for 30 minutes to 24 hours. The amount of the compound [4] used is preferably 1 to 1.2 times the molar amount of the compound [3].

(2)

Embedded image [In the formula, R ² and R ³ are the same as above. R ¹¹¹ represents unsubstituted alkyl. ]

The ester [5] can be prepared by reacting the carboxylic acid [3] with an O-alkylating agent. As the O-alkylating agent, diazoalkane (eg, diazomethane, diazoethane, etc.), trimethylsilyldiazomethane, orthoester (eg, ethyl orthoformate, ethyl orthoacetate, etc.) can be used. This reaction can usually be carried out without solvent or in the same aprotic solvent as described above. The reaction temperature is O-
It depends on the type of alkylating agent, and in the case of diazoalkane and trimethylsilyldiazomethane, it is -20 to 3
0 ° C is suitable, 100-200 ° C for orthoesters
Is appropriate. The reaction time varies depending on the kinds of the compound [3] and the O-alkylating agent and the reaction time, but is usually 1
Minutes to 24 hours is appropriate. The amount of the O-alkylating agent used is preferably 1 to 1.2 times the molar amount of the compound [3].

(3)

Embedded image [In formula, R < ² >, R ^{< 3 >} , R ^<110> is the same as the above. Y is hydroxy or halogen such as chlorine, bromine and iodine, alkoxy such as methoxy, aryloxy such as p-nitrophenoxy, alkylsulfoxy such as methanesulfoxy, arylsulfoxy such as toluenesulfoxy, imidazolyl and alkylcarboxy. Alternatively, it represents a leaving group such as arylcarboxy. ]

The ester [5] can be produced by reacting the compound [6] with the alcohol [7]. Specifically, the ester [5] is a compound [6] (Y
Is a leaving group other than hydroxy), for example, an acid halide (eg, acid chloride, acid bromide, etc.), an alkyl ester (eg, methyl ester, ethyl ester, etc.), an active ester (eg, p-nitro). Phenyl ester, p-chlorophenyl ester, etc.), imidazolide or mixed acid anhydride (eg, alkyl carbonic acid mixed acid anhydride, alkyl phosphoric acid mixed acid anhydride) and the like, or a method of appropriately reacting the alcohol [7], or a compound [6] ] (When Y is hydroxy) and a compound [7]
As a condensing agent (for example, 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide, dicyclohexylcarbodiimide, 2-chloro-N-methylpyridinium iodide, diphenylphosphoryl azide, diethylphosphoryl cyanide, triphenylphosphine-tetrachloride. Carbon etc.)
It can be produced by a method of directly binding using.

When an acid halide is used, the ester [5]
Can be produced by reacting in the same aprotic solvent as described above in the presence of the same base as described above at -20 to 100 ° C. The reaction time varies depending on the type of acid halide and the reaction temperature, but is usually 30 minutes to 24 hours.
The amount of alcohol [7] used is 1 to 1 with respect to the acid halide.
A double molar amount is preferred. The acid halide is a compound [6]
(When Y is hydroxy) and thionyl halide (thionyl chloride, thionyl bromide, etc.) without solvent or in the same aprotic solvent as described above, in the presence or absence of the same base as described above, at -20 to 100 ° C. It can be produced by reacting with. The reaction time varies depending on the type of acid halide and the reaction temperature, but is usually 30 minutes to 24 hours. The amount of thionyl halide used needs to be at least 1-fold mole with respect to compound [6] (when Y is hydroxy), and a large excess such as 10-fold mole or more can be used. When a condensing agent is used, the ester [5] can be produced by carrying out a reaction in the same aprotic solvent as described above at -20 to 100 ° C in the presence or absence of the same base as described above. The reaction time varies depending on the type of condensing agent and the reaction temperature, but is usually 30 minutes to 24 hours. The alcohol [7] and the condensing agent are preferably used in an amount of 1 to 1.2 times the molar amount of the compound [6] (when Y is hydroxy).

(4)

[Chemical 6] [In formula, R < ² >, R < ³ >, R <^11> , R < ¹² >, Y is the same as the above. ]

By reacting the compound [6] with the amine [8], an amide is obtained.

[9] can be manufactured. Specifically, amide

[9] is a compound [6] (when Y is the above-mentioned leaving group other than hydroxy), for example, an acid halide (eg, acid chloride, acid bromide, etc.), an alkyl ester (eg, methyl ester, ethyl ester). Etc.), active ester (eg p-nitrophenyl ester, p-chlorophenyl ester etc.), imidazolide or mixed acid anhydride (eg alkyl carbonic acid mixed acid anhydride, alkyl phosphoric acid mixed acid anhydride) and the like and amine [8 ]
Or a compound [6] (where Y is hydroxy) and a compound [8] as a condensing agent (for example, 1
-Ethyl-3- (3-dimethylaminopropyl) carbodiimide, dicyclohexylcarbodiimide, diisopropylcarbodiimide, benzotriazol-1-yl-tris (dimethylamino) phosphonium hexafluorophosphide salt, diphenylphosphoryl azide, propanephosphoric anhydride, etc.) Directly in the presence or absence of additives (for example, N-hydroxysuccinimide, 1-hydroxybenzotriazole, 3-hydroxy-4-oxo-3,4-dihydro-1,2,3-triazine, etc.). It can be manufactured by a bonding method.

When an acid halide is used, the amide

[9] can be produced by reacting in the same aprotic solvent as described above in the presence of the same base as described above at -20 to 100 ° C. The reaction time varies depending on the type of acid halide and the reaction temperature, but is usually 30 minutes to 24 hours. The amount of amine [8] used is preferably 1 to 1.2 times the molar amount of the acid halide. If a condensing agent is used, the amide

[9] can be produced by reacting in the same aprotic solvent as described above in the presence or absence of the same base as described above at -20 to 100 ° C. The reaction time varies depending on the type of condensing agent and the reaction temperature, but is usually 30 minutes to 24 hours. The amount of the amine [8] and the condensing agent used is the compound [6].
It is preferably 1 to 2 times the molar amount with respect to (when Y is hydroxy). It is also possible to use the amine [8] as a base in excess.

Reaction of hydroxy at the 23-position (1)

Embedded image [In the formula, R ² and X are the same as defined above. R ¹⁰ represents R ¹ excluding hydroxy. R ³⁰ represents R ³ excluding hydrogen. ]

The 23-position hydroxy compound [10] is converted to the compound [1
Compound [12] (for example, ether form, ester form, carbonic acid ester form, carbamic acid ester form) can be produced by reacting with 1]. This reaction can be carried out usually at -20 to 100 ° C in the absence of solvent or in the same aprotic solvent as described above and in the presence or absence of the same base as described above. The reaction time is compound [10] and compound [11]
Usually, 30 minutes to 24 hours is suitable, though it depends on the type and reaction temperature. The amount of compound [11] used is compound [10]
It is preferably 1 to 1.2 times the molar amount.

(2)

Embedded image

[In the formula, R ¹⁰ is the same as above. R ²⁰ represents R ² excluding hydrogen. R ⁴ and R ⁵ are the same or different and each represents an alkyl which may have a substituent, and each of the substituents is halogen, OR ⁶ , OCOR ⁶ , COO.
R ⁶ , cyano, NR ⁶ R ⁷ , cycloalkyl, halogen, alkyl, aryl optionally substituted by hydroxy or amino, and aromatic heterocyclic group optionally substituted by halogen, alkyl, hydroxy or amino To be elected. R ⁶ and R ⁷ are the same or different and are hydrogen;
Or, represents alkyl which may be substituted with hydroxy, alkoxy, amino, monoalkylamino, or dialkylamino. ]

The carbamic acid ester [14] can be produced by reacting the 23-position hydroxy derivative [13] with a carbonylating agent and subsequently with an amine [23]. As the carbonylating agent, carbonyldiimidazole, p-nitrochloroformate and the like can be used. The amount of the carbonylating agent and the amine [23] used is preferably 1 to 1.2 times the molar amount of the compound [13]. Other reaction conditions are the same as the reaction for producing the compound [12] from the compound [10] and the compound [11]. It is also possible to use the amine [23] as a base in excess.

3.3 Reaction of hydroxy at β-position (1)

Embedded image [In the formula, R ¹⁰ , R ²⁰ , R ³⁰ , and X are the same as defined above. ]

The 3β-hydroxy compound [15] is converted into the compound [1
Compound [17] (for example, ether body, ester body, carbonate ester body, carbamate ester body) can be produced by reacting with 6]. This reaction can be carried out in the same manner as the above-mentioned reaction for producing compound [12] from compound [10] and compound [11].

(2)

Embedded image [In the formula, R ⁴ , R ⁵ , R ¹⁰ and R ³⁰ are the same as defined above. ]

The carbamic acid ester [18] can be produced by reacting the hydroxy compound at the 23-position [15] with the same carbonylating agent as described above, and then reacting with the amine [23]. This reaction can be carried out in the same manner as the above-mentioned reaction for producing carbamic acid ester [14] from compound [13], carbonylating agent and amine [23].

4.3 Reaction of dihydroxy groups at β and 23 positions (1)

Embedded image Wherein R ¹ is the same as above. ]

The 3β-position and 23-position dihydroxy compound [19] is
3 by reacting with the same carbonylating agent as above
A β, 23-carbonyldioxy derivative [20] can be produced. This reaction is usually carried out without solvent or in the same aprotic solvent as described above in the presence or absence of the same base as described above.
It can be carried out at -20 to 100 ° C. The reaction time varies depending on the kinds of the compound [19] and the carbonylating agent and the reaction temperature, but is usually 30 minutes to 24 hours. The amount of the carbonylating agent used is preferably 1 to 1.2 times the molar amount of the compound [19].

(2)

Embedded image Wherein R ¹ is the same as above. R ²¹ represents acyl. ]

The 3β-position, 23-position dihydroxy compound [19] is
By reacting with carboxylic acid anhydride [21], 3β,
The 23-diacyloxy compound [22] can be produced. This reaction is generally carried out without a solvent or in the same aprotic solvent as described above in the presence or absence of the same base as described above.
It can be done at 100 ° C. The reaction time depends on the compound [19]
Although it depends on the kind of carboxylic acid anhydride [21] and the reaction temperature, 30 minutes to 24 hours is usually suitable. The amount of the carboxylic acid anhydride [21] used is 2 to 2.2 with respect to the compound [19].
A double molar amount is preferred.

In the above reactions (1) to (4), when the raw material has a substituent (for example, amino, hydroxy, carboxy, etc.) which is not desired to be reacted, the raw material is previously prepared by a known method such as benzyl, acetyl, It may be used in the reaction after protection with tert-butoxycarbonyl or the like. After the reaction, the protecting group can be removed by a known method such as catalytic reduction, alkali treatment, acid treatment and the like. R ¹¹ , R ¹² , R ² or R ^{3 of} the compound obtained in the above reaction
Is an alkyl substituted with a halogen, an alkenyl substituted with a halogen, an alkynyl substituted with a halogen, or a cycloalkyl substituted with a halogen, the halogen is OR ¹³ (R ¹³ is the same as described above by a known method. Org. Ch
em., 44, 2307 (1979)), OCOR ¹³ (R ¹³ is the same as the above. Tetrahedron Lett., 1972, 1853 etc.), NR
¹³ R ¹⁴ (R ¹³ and R ¹⁴ are the same as above. Org. Synth., V, 88
(1973)), cyano (J. Org. Chem., 25, 877 (1960)
Etc.) can be converted to. When the compound obtained in the above reaction has cyano, C
OOR ¹³ (R ¹³ is the same as above. Org. Synth., III, 557
(1955), Org. Synth., I, 27 (1941), etc.) or an aromatic heterocyclic group such as triazinyl (Org. Synth., IV, 78 (196
3) etc.) can be converted.

When the compound obtained in the above reaction has an alkoxycarbonyl, carboxy (Org. Synth., IV, 169 (1963), Org. Synth., IV,
608 (1963) etc.) can be converted. When the compound obtained in the above reaction has carboxy, alkoxycarbonyl (Org. Synth., II
I, 381 (1955), etc.). When the compound obtained in the above reaction has hydroxy, OR ¹³ (R ¹³ is the same as the above except hydrogen, by a known method.
J. Org. Chem., 44, 2307 (1979)), OCOR
¹³ (R ¹³ is the same as above. Org. Synth., IV, 263 (1963),
Org. Synth., VI, 560 (1988)). When the compound obtained in the above reaction has amino, NR ¹³ R ¹⁴ (R ¹³ and R ¹⁴ are the same as above. Org. Synth., V, 88 (1973)), pyrrole (Org. Synth., II, 219 (1943), etc.) can be converted into an aromatic heterocyclic group.

[0049]

BEST MODE FOR CARRYING OUT THE INVENTION The carboxy-containing hederagenin derivative [1] can be used for treatment as a free carboxylic acid, but can be used in the form of a pharmaceutically acceptable salt by a known method. Examples of the salt include alkali metal salts such as sodium salt and potassium salt, and alkaline earth metal salts such as calcium salt. For example, an alkali metal salt of a hederagenin derivative [1] is a hederagenin derivative [1] having a carboxy.
Can be obtained by adding 1 equivalent of sodium hydroxide or potassium hydroxide, preferably in an alcohol solvent. The alkaline earth metal salt of the hederagenin derivative [1] can be obtained by dissolving the alkali metal salt produced by the above method in water, methanol, ethanol or a mixed solvent thereof, and adding 1 equivalent of calcium chloride or the like. it can.

The hederagenin derivative [1] having an unsubstituted or substituted amino can be used for treatment as a free amine as it is, but can be used in the form of a pharmaceutically acceptable salt by a known method. Examples of salts include salts of mineral acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, acetic acid, citric acid,
Examples thereof include salts of organic acids such as tartaric acid, maleic acid, succinic acid, fumaric acid, p-toluenesulfonic acid, benzenesulfonic acid and methanesulfonic acid. For example,
The hydrochloride of the hederagenin derivative [1] can be obtained by dissolving the hederagenin derivative [1] having an unsubstituted or substituted amino group in an alcohol solution of hydrochloric acid.

The compound [1] according to the present invention can be obtained by subjecting the above reaction mixture to usual separation and purification means such as extraction, concentration,
It can be isolated and purified by using means such as neutralization, filtration, recrystallization, column chromatography and thin layer chromatography. Solvates (including hydrates) of the compound [1] or a salt thereof according to the present invention are also included in the present invention. The solvate can be usually obtained by recrystallizing the solvate from the corresponding solvent or an appropriate mixed solvent containing the corresponding solvent. For example, the hydrate of the compound [1] of the present invention can be obtained by recrystallizing the compound [1] of the present invention from hydrous alcohol.
The compound [1] according to the present invention may have a crystalline polymorphism. The crystal polymorph is also included in the present invention. The present invention also includes a method for treating nephritis, which comprises administering an effective amount of the compound represented by the formula [1] to humans and animals. The use of the compound represented by the formula [1] for the manufacture of a medicament for treating nephritis is also included in the present invention.

The compound according to the present invention has an excellent mesangial cell growth inhibitory action as shown in the test examples described below. It also has low toxicity. From this, the pharmaceutical composition of the present invention is useful as an excellent therapeutic agent for nephritis, and is effective for treating chronic glomerulonephritis, especially proliferative glomerulonephritis among nephritis. When the compound of the present invention is administered as a pharmaceutical, the compound of the present invention is used as it is or in a pharmaceutically acceptable non-toxic and inert carrier, for example, 0.1 to 99.5.
%, Preferably 0.5-90%, can be administered to animals, including humans, as a pharmaceutical composition.

As the carrier, one or more solid, semi-solid or liquid diluents, fillers and other auxiliary agents for formulation are used. Desirably, the pharmaceutical compositions are administered in dosage unit form. The pharmaceutical composition of the present invention can be administered intravenously, orally, intratissueally, topically (such as transdermally) or rectally. Needless to say, the composition is administered in a dosage form suitable for these administration methods. Oral administration is especially preferred. The dosage as a pharmaceutical composition for treating nephritis is preferably set in consideration of the patient's condition such as age and weight, administration route, nature and degree of disease, etc. The amount of the active ingredient of the compound of the present invention is generally in the range of 0.1 to 1000 mg / human, preferably in the range of 1 to 500 mg / human per day. In some cases, less may be sufficient, and conversely, higher doses may be required. It can also be administered in divided doses two to three times a day.

The present invention will be described in more detail with reference to production examples, test examples and examples relating to the compound of the present invention.

Reference Example 1 Hederagenin (3β, 23-dihydroxyolean-1
2-en-28-oic acid, Compound No. 1) 6.0 kg of dried and crushed pericarp of Sacrodus mukorossi Gaertn. Was heat-extracted twice with 60 L (liter) of methanol at 70 ° C. for 2 hours, and the extract solution was 50 times. The mixture was concentrated under reduced pressure at ℃ or below to obtain 3.88 kg of methanol extract. This methanol extract
To 300 g, add 1500 ml of 2.5% (v / v) sulfuric acid methanol solution and add 2
The mixture was heated under reflux for an hour and hydrolyzed. After hydrolysis, 5% (w / v)
The pH was adjusted to 6 to 7 with a methanol solution of potassium hydroxide, 90 g of activated carbon was added, and the mixture was refluxed for 0.5 hour. 2 g of this solution is Celite
The mixture was filtered through a homogenous layer of 1 and the filtrate was concentrated under reduced pressure to a volume of about 1/10. Distilled water (1 L) was added to the residue, which was heated and washed, then allowed to cool, the residual methanol was distilled off under reduced pressure, and the precipitate was suction filtered to obtain crude hederagenin. This crude hederagenin was washed twice with 1 L of acetonitrile and 3 times with 80 ml of ethanol to obtain 11.05 g of purified hederagenin as a white powder. Mp 317-320 ° C. Elemental analysis _{(C 30 H 48 O 4 ·} 1 / 4H 2 O ) Calculated value (%) C: 75.51 H: 10.24 Found (%) C: 75.49 H: 9.94 IR (KBr) cm ^-1 : 3455, 2946, 1698, 1464, 1389, 103
8

Reference Example 2 Sodium salt of hederagenin (sodium 3β, 23-
Dihydroxyolean-12-en-28-oate ) To the hederagenin (1.0 g) obtained in Reference Example 1 was added 1N-sodium hydroxide methanol solution (2.1 ml), and the mixture was stirred at 50 ° C for 2 hours. Insoluble matter is removed with filter paper and concentrated to obtain the desired product (934
mg). Mp 315 ° C. Elemental analysis _{(C 30 H 47 O 4 ·} Na · 3H 2 O ) Calculated value (%) C: 65.67, H : 9.74 Found (%) C: 65.71, H : 9.51

Production Example 1 2-acetoxyethyl 3β, 23 (4α) -dihydro
Xyolean-12-ene-28-oate (Compound No.
No. 2) To a solution of hederagenin (473 mg) in DMF (10 ml) was added 2-bromoethyl acetate (334 mg), and potassium hydrogen carbonate (20 mg) was added.
0 mg) was added and the mixture was stirred at 50 ° C. for 8 hours. The reaction solution was filtered and then concentrated under reduced pressure to remove DMF. The obtained oily substance was extracted with ethyl acetate and washed with saturated saline,
After drying over anhydrous magnesium sulfate, the mixture was concentrated under reduced pressure. Purification by silica gel column chromatography (ethyl acetate: n-hexane = 1: 8) gave colorless crystals of the desired product (490 mg).
Mp 158 ° C. Elemental analysis (as C ₃₄ H ₅₄ O ₆₎ Calculated (%) C: 73.08, H : 9.74 Found (%) C: 72.90, H : 9.64

Production Example 2 Benzyl 3β, 23 (4α) -dihydroxyolean
Using 12-en-28-oate (Compound No. 3) benzyl bromide, a target product was obtained in the same manner as in Production Example 1. Mp 159 ° C. Elemental analysis _{(C 37 H 54 O 4 ·} 1 / 5H 2 O ) Calculated value (%) C: 78.46, H : 9.68 Found (%) C: 78.50, H : 9.47

Production Example 3 2-Methoxyethyl 3β, 23 (4α) -dihydroxy
Siolean-12-ene-28-oate (Compound No.
4) Using 2-bromoethyl methyl ether, a target product was obtained in the same manner as in Production Example 1. Mp 161 ° C. Elemental analysis _{(C 33 H 54 O 5 ·} 1 / 4H 2 O ) Calculated value (%) C: 74.05, H : 10.26 Found (%) C: 74.01, H : 9.92

Production Example 4 2-Hydroxyethyl 3β, 23 (4α) -dihydro
Xyolean-12-ene-28-oate (Compound No.
No. 5) The compound (1.0 g) obtained in Preparation Example 1 was dissolved in methanol (100 ml), an aqueous sodium hydroxide solution (5 ml) was added, and the mixture was heated under reflux at 80 ° C., and then methanol was distilled off. The residue was extracted with ethyl acetate, dried over anhydrous magnesium sulfate,
Filtration and concentration gave colorless crystals of the desired product (880 mg). Melting point
237-238 ℃ Elemental analysis value (as C ₃₂ H ₅₂ O ₅ ) Calculated value (%) C: 74.38, H: 10.14 Measured value (%) C: 74.06, H: 9.90

Production Example 5 3-Bromopropyl 3β, 23 (4α) -dihydroxy
Siolean-12-ene-28-oate hydrochloride (compound
Product number 6) Using 1,3-dibromopropane, a target product was obtained in the same manner as in Production Example 1.

Production Example 6 3-N, N-di (2-hydroxyethyl) aminopropyl
3β, 23 (4α) -dihydroxyolean-12
-Ene-28-oate hydrochloride (Compound No. 7) The crystal (622 mg) obtained in Preparation Example 5 was dissolved in DMF (60 ml),
Diethanolamine (2 ml) was added, and the mixture was stirred at room temperature for 24 hours. The mixture was extracted with ethyl acetate, dried over anhydrous magnesium sulfate, concentrated, and purified by silica gel column chromatography (ethyl acetate: n-hexane = 3: 1). The obtained crystals (580 mg) were dissolved in 20% hydrochloric acid-methanol solution (20 ml), stirred at room temperature for 30 minutes and then concentrated to give colorless crystals of the desired product.
(572 mg) was obtained. Mp 267-268 ° C. Elemental analysis _{(C 37 H 63 NO 6 ·} HCl · 1 / 2H as ₂ O) Calculated (%) C: 66.99, H : 9.88, N: 2.11 Found (%) C: 67.22, H: 9.40, N: 2.33

Production Example 7- (1) N- (2-methoxyethyl) -3β, 23 (4α) -di
Acetoxyolean-12-ene-28-amide (compound
Item No. 8) Step 1 3β, 23 (4α) -diacetoxyolean-
12-Ene-28-oic acid hederagenin (500 mg) was dissolved in pyridine (20 ml), acetic anhydride (2 ml) was added, and the mixture was stirred at 80 ° C for 1 hr. 1 in the reaction solution
N hydrochloric acid (5 ml) was added dropwise and the mixture was stirred for 10 minutes. The reaction solution was extracted three times with chloroform (50 ml), the extract was dried over anhydrous magnesium sulfate, and the solvent was distilled off. The residue was subjected to silica gel column chromatography (n-hexane: ethyl acetate = 4:
Purification by 1) gave the desired product (580 mg) as a white solid.

Step 2 N- (2-methoxyethyl) -3
β, 23 (4α) -diacetoxyolean-12-ene
Thionyl chloride (20 ml) was added to the compound (570 mg) obtained in -28-amide step 1 and the mixture was stirred at 70 ° C for 1.5 hours, and unreacted thionyl chloride was evaporated under reduced pressure to give a yellow oil. Add this to methylene chloride (5
This was dissolved in 0 ml), 2-methoxyethylamine (2 ml) was added, and the mixture was stirred at room temperature for 12 hours. The reaction mixture was washed twice with 1N hydrochloric acid (50 ml), the organic layer was dried over anhydrous magnesium sulfate and the solvent was distilled off to give a yellow solid. This was purified by silica gel column chromatography (n-hexane: ethyl acetate = 4: 1 → 1: 1) to obtain the desired product (565 mg) as a white solid.

Production Example 7- (2) N- (2-methoxyethyl) -3β, 23 (4α) -di
Acetoxyolean-12-ene-28-amide (compound
Item No. 8) Step 1 3β, 23 (4α) -diacetoxyolean-
12-Ene-28-oic acid hederagenin (1.2 g) was dissolved in pyridine (15 ml), acetic anhydride (4 ml) was added, and the mixture was stirred at 70 ° C for 1 hr. 1 in the reaction solution
N hydrochloric acid (30 ml) was added dropwise and the mixture was stirred for 10 minutes. The reaction mixture was extracted with ether (20 ml) three times, the extract was dried over anhydrous magnesium sulfate, and the solvent was evaporated. The residue was subjected to silica gel column chromatography (n-hexane / ethyl acetate =
The product was purified by 5: 1) to obtain the desired product (1.35 g) as a white solid.

Step 2 N- (2-methoxyethyl) -3
β, 23 (4α) -diacetoxyolean-12-ene
Thionyl chloride (20 ml) was added to the compound (1.2 g) obtained in Step -28-amide step 1 and the mixture was stirred at 80 ° C for 1 hr, and unreacted thionyl chloride was distilled off under reduced pressure to obtain a yellow oily substance. 2-Methoxyethylamine (330 mg) was added thereto, and the mixture was stirred at room temperature for 1 hour. It was extracted with ether, washed with saturated brine, dried over anhydrous magnesium sulfate, and the solvent was distilled off to obtain the desired product (950 m
g) was obtained as a yellow solid.

Production Example 8- (1) N- (2-methoxyethyl) -3β, 23 (4α) -di
Hydroxyolean-12-ene-28-amide (compound
No. 9) The compound (155 mg) obtained in Production Example 7- (1) was treated with methanol (30 mg).
ml) and add 1N aqueous sodium hydroxide solution (10 ml),
The mixture was refluxed for 1 hour. Methanol was evaporated under reduced pressure, and the residue was extracted with ethyl acetate (30 ml) three times. The extract was dried over anhydrous magnesium sulfate, the solvent was evaporated, and the residue was crystallized from n-hexane / ethyl acetate to give the desired product (140 mg) as a white powder. Melting point 127-129 ℃ Elemental analysis value (as C ₃₃ H ₅₅ NO ₄・ 3 / 5H ₂ O) Calculated value (%) C: 73.32 H: 10.48 N: 2.59 Measured value (%) C: 73.21 H: 10.24 N: 2.85

Production Example 8- (2) N- (2-methoxyethyl) -3β, 23 (4α) -di
Hydroxyolean-12-ene-28-amide (compound
No. 9) The compound (950 mg) obtained in Production Example 7- (2) was treated with methanol (35 mg).
ml) and add 1N aqueous sodium hydroxide solution (15 ml),
The mixture was stirred at room temperature for 1 hour. Methanol was distilled off under reduced pressure,
The residue was extracted with chloroform. The extract was dried over anhydrous magnesium sulfate, the solvent was evaporated, and the residue was recrystallized from ethyl acetate to obtain the desired product (830 mg) as white crystals.
Mp 142-143 ° C. Elemental analysis _{(C 33 H 55 NO 4 ·} 3 / 5H 2 O ) Calculated value (%) C: 73.32 H: 10.48 N: 2.59 Found (%) C: 73.14 H: 10.25 N: 2.91

Production Example 9 N- (2-methoxyethyl) -3β, 23 (4α) -di
Hydroxyolean-12-en-28-amide separately
Production method (Compound No. 9) hederagenin (100 mg), 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (84 mg), 1-
Hydroxy-1H-benzotriazole (67 mg) and 2-methoxyethylamine (33 mg) were suspended in DMF (2 ml).
The mixture was stirred at ℃ for 2 hours, cooled to room temperature, and concentrated under reduced pressure. The residue was extracted with ethyl acetate, washed with saturated brine and 1N hydrochloric acid, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate: n-hexane = 1: 10 → 1: 5 → 1: 1) to obtain colorless crystals of the desired product (92 mg). The physical property values corresponded to the values in Production Example 8- (2).

Production Example 10 N- (p-fluorophenyl) -3β, 23 (4α)-
Diacetoxyolean-12-ene-28-amide
Compound No. 10) Using p-fluoroaniline, a target product was obtained in the same manner as in Production Example 7- (1).

Production Example 11 N- (p-fluorophenyl) -3β, 23 (4α)-
Dihydroxyolean-12-ene-28-amide
Compound No. 11) Using the compound obtained in Production Example 10, the desired product was obtained in the same manner as in Production Example 8- (1). Melting point 216-217 ℃

Production Example 12 N- [2- (N, N-diethylamino) ethyl] -3
β, 23 (4α) -diacetoxyolean-12-ene
-28-amide (Compound No. 12) 2- (N, N-diethylamino) ethylamine was used to obtain the desired product in the same manner as in Production Example 7- (1).

Production Example 13 N- [2- (N, N-diethylamino) ethyl] -3
β, 23 (4α) -dihydroxyolean-12-ene
-28-amide (Compound No. 13) Using the compound obtained in Production Example 12, the target compound was obtained in the same manner as in Production Example 8- (1). Melting point 228-230 ℃ Elemental analysis value (as C ₃₆ H ₆₂ N ₂ O ₃ ) Calculated value (%) C: 75.74, H: 10.95, N: 4.91 Measured value (%) C: 75.74, H: 10.82, N: 5.03

Production Example 14 N- [2- (N, N-diethylamino) ethyl] -3
β, 23 (4α) -dihydroxyolean-12-ene
-28-amide hydrochloride (Compound No. 14) The compound obtained in Preparative Example 13 was treated with 20% hydrochloric acid in methanol (20 ml).
And was stirred at room temperature for 30 minutes and then concentrated to obtain the desired product. Mp 263-265 ° C. Elemental analysis _{(C 36 H 62 N 2 O} 3 · HCl · 5 / 2H 2 O ) Calculated value (%) C: 66.28, H : 10.51, N: 4.29 Found value (%) C: 66.17, H: 10.30, N: 4.56

Production Example 15 N- [3- [N, N-di (2-hydroxyethyl) ami]
No] propyl] -3β, 23 (4α) -diacetoxyo
Rean-12-ene-28-amide (Compound No. 15) Using 3- [N, N-di (2-hydroxyethyl) amino] propylamine, the target compound was obtained in the same manner as in Production Example 7- (1). Obtained.

Production Example 16 N- [3- [N, N-di (2-hydroxyethyl) ami]
No] propyl] -3β, 23 (4α) -dihydroxyo
Lean-12-ene-28-amide (Compound No. 16) Using the compound obtained in Production Example 15, the target compound was obtained in the same manner as in Production Example 8- (1). Melting point 136-139 ℃ Elemental analysis value (as C ₃₇ H ₆₄ N ₂ O ₅ 1 / 2H ₂ O) Calculated value (%) C: 71.00, H: 10.47, N: 4.48 Measured value (%) C: 70.90, H: 10.26, N: 4.54

Production Example 17 N- [3- [N, N-di (2-hydroxyethyl) ami]
No] propyl] -3β, 23 (4α) -dihydroxyo
Lean-12-ene-28-amide hydrochloride (Compound No.
17) Using the compound obtained in Production Example 16 and in the same manner as in Production Example 14, an intended product was obtained. Mp 195 ° C. Elemental analysis _{(C 37 H 64 N 2 O} 5 · HCl · 5 / 3H 2 O ) Calculated value (%) C: 65.03, H : 10.08, N: 4.10 Found (%) C: 65.01, H: 9.96, N: 4.19

Production Example 18 N-ethoxycarbonylmethyl-3β, 23 (4α)-
Diacetoxyolean-12-ene-28-amide
Compound No. 18) Production Example 7-using glycine ethyl ester hydrochloride
The target product was obtained in the same manner as in (1).

Production Example 19 N-carboxymethyl-3β, 23 (4α) -dihydro
Xyolean-12-ene-28-amide (Compound number
19) Using the compound obtained in Production Example 18, a target product was obtained in the same manner as in Production Example 8- (1). Melting point 187-190 ℃ Elemental analysis value (as C ₃₂ H ₅₁ NO ₅・ 4 / 5H ₂ O) Calculated value (%) C: 70.63, H: 9.74, N: 2.57 Measured value (%) C: 70.51, H: 9.38, N: 2.60

Production Example 20 N-ethoxycarbonylmethyl-3β, 23 (4α)-
Dihydroxyolean-12-ene-28-amide
Compound No. 20) The compound (1.1 g) obtained in Production Example 19 was converted into 17% ethanol (40 m
After being dissolved in l) and heated at 80 ° C. for 2 hours, ethanol was distilled off under reduced pressure. The residue was extracted with chloroform, dried over anhydrous magnesium sulfate, concentrated, and purified by silica gel column chromatography (chloroform: methanol = 9: 1) to obtain 1.1 g of the desired product as colorless crystals. Melting point 129-132 ℃ Elemental analysis value (as C ₃₄ H ₅₅ NO ₅・ 2 / 5H ₂ O) Calculated value (%) C: 72.28, H: 9.95, N: 2.48 Measured value (%) C: 72.29, H: 9.87, N: 2.61

Production Example 21 N- (3-methoxycarbonylpropyl) -3β, 23
(4α) -diacetoxyolean-12-ene-28-
Amide (Compound No. 21) Using 4-amino-n-butyric acid methyl ester hydrochloride,
The target product was obtained in the same manner as in Production Example 7- (1).

Production Example 22 N- (3-carboxypropyl) -3β, 23 (4α)
-Dihydroxyolean-12-ene-28-amide
(Compound No. 22) Using the compound obtained in Production Example 21, a target product was obtained in the same manner as in Production Example 8- (1). Melting point 203 ° C Elemental analysis value (as C ₃₄ H ₅₅ NO ₅ · H ₂ O) Calculated value (%) C: 70.92, H: 9.98, N: 2.43 Measured value (%) C: 71.14, H: 10.01, N: 2.49

Production Example 23 N- (3-ethoxycarbonylpropyl) -3β, 23
(4α) -dihydroxyolean-12-ene-28-
Amide (Compound No. 23) Using the compound obtained in Production Example 22 and in the same manner as in Production Example 20, an intended product was obtained. Melting point 108-110 ℃ Elemental analysis value (as C ₃₆ H ₅₉ NO ₅・ 3 / 10H ₂ O) Calculated value (%) C: 73.13, H: 10.16, N: 2.37 Measured value (%) C: 72.91, H: 10.14, N: 2.35

Production Example 24 N- (2-cyanoethyl) -3β, 23 (4α) -dia
Cetoxyolean-12-ene-28-amide (compound
No. 24) Production Example 7-using 3-aminopropionitrile
The target product was obtained in the same manner as in (1). TLC (Merck K
ieselgel 60 F ₂₅₄ , ethyl acetate: n-hexane = 1: 1) Rf =
0.13

Production Example 25 N- (2-cyanoethyl) -3β, 23 (4α) -dihi
Droxyolean-12-ene-28-amide (compound
No. 25) Using the compound obtained in Production Example 24, an intended product was obtained in the same manner as in Production Example 8- (1).

Production Example 26 N- [2- (2,4-diamino-1,3,5-triazide]
N-6-yl) ethyl] -3β, 23 (4α) -dihydr
Roxyolean-12-ene-28-amide (Compound No.
No. 26) The compound (550 mg) obtained in Preparation Example 25 was replaced with dicyandiamide (210 m
g), potassium hydroxide (140 mg) in methyl cellosolve (50 ml)
The suspension was heated under reflux at 125 ° C. for 2 hours, the solvent was distilled off under reduced pressure, and the residue was purified by silica gel column chromatography (chloroform → chloroform: methanol = 9: 1).
After washing the obtained crystals with hot water, collect the crystals in a plate,
The desired colorless crystals were obtained. Mp 214 ° C. Elemental analysis _{(C 35 H 56 N 6 O} 3 · 3 / 2H 2 O ) Calculated value (%) C: 66.11, H : 9.35, N: 13.22 Found (%) C: 65.91, H : 9.16, N: 13.12

Production Example 27 N- (3-methoxypropyl) -3β, 23 (4α)-
Diacetoxyolean-12-ene-28-amide
Compound No. 27) Production Example 7 using 3-aminopropyl methyl ether
The target product was obtained in the same manner as in (1).

Production Example 28 N- (3-methoxypropyl) -3β, 23 (4α)-
Dihydroxyolean-12-ene-28-amide
Compound No. 28) Using the compound obtained in Production Example 27, the desired product was obtained in the same manner as in Production Example 8- (1). Mp 83 ° C. Elemental analysis _{(C 34 H 57 NO 4 ·} 1 / 4H 2 as O) Calculated (%) C: 74.48, H : 10.57, N: 2.55 Found (%) C: 74.49, H : 10.36
N: 2.90

Production Example 29 Benzyl 3β-hydroxy-23 (4α) -benzyl
Olean-12-ene-28-oate (Compound No. 2
9) The compound (3.0 g) obtained in Preparation Example 2 was dissolved in DMF (40 ml),
Sodium hydride (60%, 552 mg) was added, and the mixture was stirred at 0 ° C for 1 hr. Benzyl chloride (1.7 g) was added to the reaction solution, and the mixture was stirred at room temperature for 2 hours. Water was added to the reaction solution, which was then extracted with ethyl acetate, dried over anhydrous magnesium sulfate, filtered, and concentrated. The residue was purified by silica gel column chromatography (ethyl acetate: n-hexane = 1: 20) to obtain the desired product (3.4 g).

Production Example 30 Benzyl 3β-ethoxycarbonylmethylcarbamoy
Luoxy-23 (4α) -benzylolean-12-d
N-28-oate (Compound No. 30) The compound (3.4 g) obtained in Production Example 29, carbonyldiimidazole (3.0 g), and pyridine (50 ml) were stirred at 80 ° C. for 20 hours.
Glycine ethyl ester hydrochloride (4.3 g) and 4-dimethylaminopyridine (3.8 g) were added, and the mixture was heated with stirring at 80 ° C for 20 hours, extracted and concentrated to obtain the desired product.

Production Example 31 3β-Ethoxycarbonylmethylcarbamoyloxy-
23 (4α) -hydroxyolean-12-ene-28
-Oic acid (Compound No. 31) The compound obtained in Preparation Example 30 was dissolved in methanol (200 ml) to give 5
% Palladium carbon (500 mg) was added, and the mixture was stirred under a hydrogen atmosphere for 24 hours, filtered and concentrated to obtain the desired product. Mp 196-197 ° C. Elemental analysis _{(C 35 H 55 NO 7 ·} 1 / 4H 2 as O) Calculated (%) C: 69.33, H : 9.23, N: 2.31 Found (%) C: 69.16, H : 9.04, N: 2.33

Production Example 32 Benzyl 3β- (2-methoxyethyl) carbamoyl
Oxy-23 (4α) -benzylolean-12-ene
-28-oate (Compound No. 32) Using 2-methoxyethylamine, colorless crystals of the desired product were obtained in the same manner as in Production Example 30.

Production Example 33 3β- (2-methoxyethyl) carbamoyloxy-2
3 (4α) -hydroxyolean-12-ene-28-
Oic acid (Compound No. 33) Using the compound obtained in Production Example 32, colorless crystals of the target substance were obtained in the same manner as in Production Example 31. Melting point 183-184 ℃ (recrystallized from ethyl acetate) Elemental analysis value (as C ₃₄ H ₅₅ NO ₆ ) Calculated value (%) C: 71.17, H: 9.66, N: 2.44 Measured value (%) C: 71.02, H : 9.61, N: 2.56

Production Example 34 3β, 23 (4α) -Carbonyldioxyolean-1
2-Ene-28-oic acid (Compound No. 34) hederagenin (473 mg) was dissolved in pyridine (10 ml), carbonyldiimidazole (195 mg) was added, and the mixture was heated at 50 ° C. for 1 hour. The mixture was extracted with ethyl acetate, the organic layer was washed with 1N-hydrochloric acid, dried over anhydrous magnesium sulfate, and concentrated. The residue was purified by silica gel column chromatography (ethyl acetate: n-hexane = 1: 5) to obtain the desired product (423 mg). Melting point 214-215 ℃ Elemental analysis value (as C ₃₁ H ₄₆ O ₅ ) Calculated value (%) C: 74.61, H: 9.32 Measured value (%) C: 74.36, H: 9.05

Production Example 35 2-acetoxyethyl 3β, 23 (4α) -carbonyl
Ludioxyolean-12-ene-28-oate
Compound No. 35) Using the compound obtained in Production Example 1, a target product was obtained in the same manner as in Production Example 34. Mp 112 ° C. Elemental analysis (as C ₃₅ H ₅₂ O ₇₎ Calculated (%) C: 71.89, H : 8.96 Found (%) C: 71.68, H : 8.86

The results of the pharmacological tests showing the usefulness of the representative examples of the active ingredient according to the present invention are shown below.

Test Example 1 Anti-Thy-1 Antibody Nephritis Action on Rats Anti-Thy-1 antibody nephritis is a glomerulonephritis model that occurs when Thy-1 antigen present as a membrane protein of mesangial cells reacts with the antibody thereto. . This model shows mesangial cell injury and mesangial cell proliferative lesions. Most of human chronic glomerulonephritis is proliferative glomerulonephritis whose main lesions are proliferation of mesangial cells and increase of substrate, but anti-Thy-1 antibody nephritis is human proliferative glomerulonephritis. A model of mesangial cell proliferative nephritis (Ishizaki et al., Acta. Patho
l. Jpn., 36 , 1191 (1986)).

(1) Experimental Animal Rats were used. (2) Experimental materials-Preparation of anti-Thy-1 antibody Preparation of the antibody was carried out according to the method of Ishizaki et al. That is, an adjuvant suspension of rat thymocytes was prepared, and this was subcutaneously immunized into a rabbit. After booster immunization twice, blood was collected and the obtained serum was inactivated and absorbed to obtain antiserum (anti-Thy-1 antibody).

(3) Experimental method A fixed amount of antiserum was intravenously injected from the tail vein of rats to induce nephritis. From the day after injection, add suspension of test drug 1
Orally administered once daily for 7 days. On the 8th day after the start of administration, dissection was performed and the kidney was removed. After fixing the kidney with 10% phosphate buffered formalin, a paraffin section was prepared according to a conventional method, stained with periodate Schiff, and examined microscopically. As a histopathological evaluation method, the number of intraglomerular cells (mesangial cells) in each tissue sample was counted using an optical microscope. Statistical test is unpaired t-test
Method was used. Table 1 shows the results.

[0100]

[Table 1]

From the above results, the compound according to the present invention is
It is clear that anti-Thy-1 antibody has a mesangial cell proliferation inhibitory effect on rat nephritis.

Test Example 2 Effect of MRL / lpr Mouse on Nephritis MRL / lpr mouse is a lupus nephritis model in which proliferative glomerulonephritis spontaneously occurs secondarily due to autoimmune abnormality. Using this model nephritis, a 10-week long-term oral administration test of the test drug was conducted. (1) Laboratory animals Purchased from the Jackson Laboratory,
MRL / lpr mice subcultured and maintained at Nippon Shinyaku Co., Ltd. were used.

(2) Experimental Method A suspension of the test drug was orally administered once a day for 10 weeks from the age of 8 weeks and for 6 days per week. During the administration period, a semi-quantitative test for urinary protein (test strip method) was performed. After the end of the administration period, dissection was performed and the kidney was removed. After fixing the kidney with 10% phosphate buffered formalin, a paraffin section was prepared according to a conventional method, stained with periodate Schiff's stain, and examined microscopically. As a histopathological evaluation method, the glomeruli in each tissue specimen are examined using an optical microscope, and the method of Berden (J. Immunology, 130,
1699-1705 (1983)), the degree of lesion was classified into grades, and the appearance rate (%) of each grade was calculated for each group. Statistical significance was performed using the Mann Whitney rank sum test. The results are shown in Tables 2 and 3.

[0104]

[Table 2]

[0105]

[Table 3]

From the above results, the compound according to the present invention is
It is clear that it has an inhibitory effect on urinary protein and renal tissue lesion progression in MRL / lpr mice, which is a model of lupus nephritis.

Test Example 3 Inhibitory Effect on Cultured Mesangial Cell Growth (1) Experimental Materials and Experimental Methods Glomeruli were isolated from rat kidneys by the sieving method,
This was subcultured to obtain cultured mesangial cells. 24 hours after the mesangial cells were seeded on the plate, LPS (lipopolysaccharide) -stimulated macrophage culture supernatant (solution containing growth stimulating factor) and the test drug were added and cultured for 72 hours. After completion of the culture, the mesangial cells were fixed and stained (crystal violet), and the absorbance was measured.
The mesangial cell growth inhibitory activity of the test drug is the inhibition rate against the absorbance of control mesangial cells cultured under the same conditions.
It was evaluated by (%). Table 4 shows the results.

[0108]

[Table 4]

From the above results, the compound according to the present invention is
It is apparent that it has a growth inhibitory activity on cultured mesangial cells.

Test Example 4 Acute toxicity To a mouse, a suspension of the test drug was orally administered at a dose of 2 g / kg, and general symptoms were observed until one week later. The compound of Compound No. 1 and the compound of Compound No. 9 were used as test drugs. As a result, there were no deaths and no abnormal findings were observed in any of the administration groups.

Example 1 Tablet (oral tablet) Formulation 1 tablet 80 mg In compound No. 1 compound 5.0 mg Corn starch 46.6 mg Crystalline cellulose 24.0 mg Methylcellulose 4.0 mg Magnesium stearate 0.4 mg This mixed powder was tablet-molded to give an internal tablet. And

Example 2 Tablet (oral tablet) Formulation 1 tablet 80 mg Compound No. 2 5.0 mg Corn starch 46.6 mg Crystalline cellulose 24.0 mg Methylcellulose 4.0 mg Magnesium stearate 0.4 mg This mixed powder was tableted to form an internal tablet. To do.

Example 3 Tablet (oral tablet) Formulation 1 tablet 80 mg In compound No. 9 5.0 mg Corn starch 46.6 mg Crystalline cellulose 24.0 mg Methylcellulose 4.0 mg Magnesium stearate 0.4 mg This mixed powder was tableted to form an internal tablet. To do.

[0114]

INDUSTRIAL APPLICABILITY As described above, the active ingredient according to the present invention has an excellent mesangial cell proliferation inhibitory effect, is effective in a model of nephritis in animals, and has low toxicity, and is therefore useful in the treatment of nephritis. .

Claims (10)

[Claims] 1. A pharmaceutical composition for treating nephritis, comprising a hederagenin derivative represented by the following formula [1], a pharmaceutically acceptable salt thereof, or a solvate of any of them as an active ingredient. Embedded image In the formula, Me represents CH ₃ . R ¹ is OR ¹¹ or NR ¹¹ R
Represents ¹² . R ¹¹ and R ¹² are the same or different and are hydrogen,
Alkyl which may have a substituent, cycloalkyl which may have a substituent, alkenyl which may have a substituent, alkynyl which may have a substituent, and which has a substituent. Represents an optionally substituted aryl or an aromatic heterocyclic group optionally having a substituent. R ² , R ³
Are the same or different and are hydrogen, alkyl optionally having substituent (s), cycloalkyl optionally having substituent (s), alkenyl optionally having substituent (s), optionally substituent (s) Good alkynyl, optionally substituted acyl, optionally substituted monoalkylcarbamoyl, optionally substituted dialkylcarbamoyl, or optionally substituted alkoxy It represents carbonyl, or R ² and R ³ together represent carbonyl. 2. The substituents for R ¹¹ , R ¹² , R ² , and R ³ are halogen, OR ¹³ , OCOR ¹³ , and COO, respectively.
R ¹³ , cyano, NR ¹³ R ¹⁴ , cycloalkyl,
The hederagenin derivative or its pharmaceutical according to claim 1, which is selected from aryl optionally substituted with halogen, alkyl, hydroxy or amino, and aromatic heterocyclic group optionally substituted with halogen, alkyl, hydroxy or amino. A pharmaceutical composition for the treatment of nephritis, which comprises a salt or a solvate of any of them as an active ingredient. Here, R ¹³ and R ¹⁴ are the same or different and are hydrogen, or hydroxy, alkoxy, amino, monoalkylamino,
Alternatively, it represents alkyl optionally substituted with dialkylamino. 3. R ¹ is hydroxy, acyloxyalkoxy, phenylalkoxy, alkoxyalkoxy, hydroxyalkoxy, haloalkoxy, di (hydroxyalkyl) aminoalkoxy, (alkoxyalkyl).
Amino, halogen-substituted phenylamino, (dialkylaminoalkyl) amino, [di (hydroxyalkyl) aminoalkyl] amino, (carboxyalkyl) amino, (alkoxycarbonylalkyl) amino, (cyanoalkyl) amino or diamino-substituted triazinylalkyl A drug for treating nephritis, which comprises amino, R ² is hydrogen, and R ³ is hydrogen, and the hederagenin derivative or a pharmaceutically acceptable salt thereof or a solvate of any of them is used as an active ingredient. Composition. 4. R ¹ is hydroxy, (alkoxyalkyl) amino, halogen-substituted phenylamino, (dialkylaminoalkyl) amino, [di (hydroxyalkyl) aminoalkyl] amino, (alkoxycarbonylalkyl) amino or (cyanoalkyl). A medicament for treating nephritis, which comprises amino, R ² is acetoxy, and R ³ is acetoxy, and a hederagenin derivative or a pharmaceutically acceptable salt thereof or a solvate of any of them is used as an active ingredient. Composition. 5. The hederagenin derivative or its pharmaceutical composition according to claim 1, wherein R ¹ is phenylalkoxy, R ² is hydroxy, (alkoxycarbonylalkyl) carbamoyl or (alkoxyalkyl) carbamoyl, and R ³ is phenylalkyl. A pharmaceutical composition for treating nephritis, which comprises a pharmaceutically acceptable salt or a solvate thereof as an active ingredient. 6. The hederagenin derivative or a pharmaceutically acceptable derivative thereof according to claim 1, wherein R ¹ is hydroxy, R ² is (alkoxycarbonylalkyl) carbamoyl or (alkoxyalkyl) carbamoyl, and R ³ is hydrogen. A pharmaceutical composition for treating nephritis, which comprises a salt or a solvate of any of them as an active ingredient. 7. A hederagenin derivative or a pharmaceutically acceptable salt thereof or a solvent thereof according to claim 1, wherein R ¹ is hydroxy or acyloxyalkoxy, and R ² and R ³ together are carbonyl. A medicinal composition for treating nephritis, which comprises a Japanese product as an active ingredient. 8. A hederagenin derivative or a pharmaceutically acceptable derivative thereof according to claim 1, wherein R ¹ is hydroxy, 2-methoxyethylamino or 2-acetoxyethoxy, R ² is hydrogen and R ³ is hydrogen. A pharmaceutical composition for treating nephritis, which comprises a salt or a solvate of any of them as an active ingredient. 9. A pharmaceutical composition for treating chronic glomerulonephritis, which comprises the hederagenin derivative according to any one of claims 1 to 8, a pharmaceutically acceptable salt thereof, or a solvate of any of them as an active ingredient. 10. A pharmaceutical composition for treating proliferative glomerulonephritis, which comprises the hederagenin derivative according to any one of claims 1 to 8, a pharmaceutically acceptable salt thereof, or a solvate of any of them as an active ingredient.