JP2001064176A - Fat accumulation inhibitor containing amide-based compound as active ingredient - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a fat accumulation inhibitor capable of suppressing the accumulation of fat in fat-cells, and thereby capable of suppressing the increasing of fat tissue, and useful for the prevention or treatment of various diseases associated with the increasing of fat tissue by formulating a specific compound, its prodrug or a pharmaceutically permissible salt of such a compound. SOLUTION: This inhibitor contains a compound expressed by formula I [R1 is H, a 1-4C alkyl or a 1-8C alkanoyl; R2 is H or a 1-4C cyano or the like; R3 is H or a 1-4C alkyl; R4 is H, a halogen or the like; R5 is H, a halogen, OH or the like; R6 is H, a halogen, a 3-6C alkyl or the like; m is 1 or 2; n is 1 or 4; A is formula II (R7 is H, a 1-4C alkyl or the like; R8 and R9 form a saturated 5- or 6-membered ring or the like when they are adjacent to each other) or the like; X is O or S; Y is O, S, SO or the like], its prodrug or a pharmaceutically permissible salt of such a compound. Further, the daily dose for an adult is, for example, usually 1 mg to 1 g. This may be adiministered in one or several divided doses.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a fat accumulation inhibitor.

[0002]

2. Description of the Related Art Fat accumulation in fat cells in the abdomen, thighs, buttocks, mesentery, epididymis, peri-kidney, breast, etc. is accompanied by, for example, an increase in visceral adipose tissue and body fat. Obesity and impaired glucose tolerance [Journal of Clinical Investigation,
72, 1150 (1983)], Diabetes [National Dia
betes Data Group: Diabetes in America. Bethesda, M
D, US Dept. of Health and Human Services, (19
85), Diabetes Care, 19, 613 (1996),
Diabete & Metabolisme, Vol. 20, p. 375 (199
4), Obesity: Advances in Understanding and Treat
ment, Published by IBC Biomedical Library. Chapter
3.1, (1996)], hyperglyceridemia, hyperlipidemia, hypertension [Journal of Clinical Investigation, 72]
Volume 1150 (1983)], Coronary artery disease [Diabete & M
etabolisme, 20, 375 (1994)], and is known to induce diseases such as obstructive arteriosclerosis [WHO Exper
t Committee on Diabetes Mellitus. Second report. W
HO Tech Rep 646 Geneva: WorldHealth Organization
(1980)].

[0003]

Therefore, fat accumulation in fat cells is suppressed, and thus increase in adipose tissue is suppressed.
There is a strong need for the development of drugs useful for the prevention and treatment of various diseases associated with adipose tissue enlargement.

[0004]

Means for Solving the Problems Under these circumstances, the present inventors have conducted intensive studies and have found that a compound represented by the following general formula (1), a prodrug thereof, or a pharmaceutically acceptable salt thereof is provided. (Hereinafter sometimes abbreviated as the compound of the present invention) has been found to have an excellent effect of suppressing fat accumulation in adipocytes, leading to the present invention. That is, the present invention
The present invention relates to the following invention. [1] General formula (1):

Embedded image [In the formula, R ¹ represents a hydrogen atom, an alkyl group having 1 to 4 carbon atoms, or an alkanoyl group having 1 to 8 carbon atoms. R ²
Represents a hydrogen atom, an alkyl group having 1 to 4 carbon atoms or a cyano group. R ³ represents a hydrogen atom or an alkyl group having 1 to 4 carbon atoms. R ⁴ independently represents a hydrogen atom, a halogen atom or an alkyl group having 1 to 4 carbon atoms when there are a plurality of them. R ⁵ is a hydrogen atom, a halogen atom, a hydroxyl group, an amino group, a cycloalkyl group having 3 to 8 carbon atoms, an aryl group having 6 to 10 carbon atoms, an alkoxy-substituted alkyl group having 2 to 8 total carbon atoms, and having a total number of carbon atoms. 2-8 alkylthio substituted alkyl group, a haloalkylthio-substituted alkyl group having the total carbon number of 2 to 8, wherein 1 to 8 carbon atoms: groups represented by -NHR ^51, the total number of carbon atoms is 2 to 16 of the formula: -NR ⁵² group represented by R ^53, wherein 1 to 8 carbon atoms: groups represented by -NHCOR ^54, wherein from 1 to 8 carbon atoms: -N ^{= CR} 54 ^{R 5 5,} a group represented by Formula having a total carbon number of 2 to 8: -N = CHNHR
A group represented by ⁵² , a formula having a total carbon number of 3 to 8: -N = C
(CH ₃ ) a group represented by NHR ⁵² , wherein the total number of carbon atoms is 3 to 1
Formula 6: -N = CHNR ⁵² R ⁵³ , a formula having a total of 4 to 16 carbon atoms: -N = C (CH ₃ ) NR ⁵² R
A group represented by ⁵³ or a formula having 1 to 8 carbon atoms: -NHS
A group represented by O ₂ R ⁵¹ , or a formula: —R ⁵⁶ , —OR
⁵⁶ , —SR ⁵⁶ , —SOR ⁵¹ , —SO ₂ R ⁵¹ or —OSO ₂ R ⁵¹ (R ⁵¹ has 1 carbon atom)
Represents an alkyl group having 1 to 8 carbon atoms or a haloalkyl group having 1 to 8 carbon atoms, R ⁵² and R ⁵³ each independently represent an alkyl group having 1 to 8 carbon atoms, and R ⁵⁴ and R ⁵⁵ are
Each independently represents a hydrogen atom or an alkyl group having 1 to 8 carbon atoms, and R ⁵⁶ represents an alkyl group having 1 to 8 carbon atoms,
C1-C8 haloalkyl group, C2-C8 alkenyl group, C2-C8 haloalkenyl group, C2-C2
8 alkynyl group or an alkynylalkenyl group having 4 to 8 carbon atoms). When there are a plurality of R ⁶ , each independently represents a hydrogen atom, a halogen atom, a cycloalkyl group having 3 to 6 carbon atoms or an alkoxy-substituted alkyl group having 2 to 3 carbon atoms, or a formula: —R
⁶¹ , -OR ⁶¹ , -SR ⁶¹ , -SOR ⁶² , -SO
₂ R ⁶² , a group represented by —OSO ₂ R ⁶² (in the formula, R
⁶¹ is an alkyl group having 1 to 3 carbon atoms, a haloalkyl group having 1 to 3 carbon atoms, an alkenyl group having 2 to 6 carbon atoms,
6 represents a haloalkenyl group or an alkynyl group having 2 to 6 carbon atoms, and R ⁶² represents an alkyl group having 1 to 3 carbon atoms or a haloalkyl group having 1 to 3 carbon atoms). When R ⁵ and R ⁶ are adjacent to each other, they are bonded at both ends to form 5
May form a six-membered ring or a six-membered ring, and these rings are saturated or unsaturated, and have an oxygen atom or a sulfur atom
2 to 2 halogen atoms or 1 to 4 carbon atoms
May be substituted with an alkyl group. m represents an integer of 1 or 2. n represents an integer of 1 to 4. A is the formula:

Embedded imageRepresents a group represented by R⁷, R¹⁰, R¹¹And R
¹²Are each independently a hydrogen atom, an alkyl group having 1 to 4 carbon atoms.
Represents a alkyl group or a haloalkyl group having 1 to 4 carbon atoms.
R⁸And R⁹Are each independently a hydrogen atom or
A halogen atom or a formula: —R⁸¹Or -OR⁸¹
(R⁸¹Is an alkyl group having 1 to 6 carbon atoms,
A haloalkyl group, a cycloalkyl group having 3 to 6 carbon atoms or
Represents an optionally substituted phenyl group)
Represents a group. Also, R⁸And R⁹When both are adjacent, both ends
To form a saturated 5-membered ring or a saturated 6-membered ring
May form a ring, and these rings have 1 to 4 carbon atoms.
It may be substituted with an alkyl group. X and Z are
Each independently represents an oxygen atom or a sulfur atom. Y is
An oxygen atom or a sulfur atom, or a formula: -SO-, -S
O₂-, -CH ₂-, -OCH₂-Or -NR¹³
− (R¹³Represents a hydrogen atom or an alkyl group).
Represents a group. Ring B is a benzene ring, a pyridine ring or
Represents a cyclohexane ring. Or a compound represented by
Prodrugs or their pharmaceutically acceptable salts
A fat accumulation inhibitor containing. [2] A is the formula:

Embedded image The fat accumulation inhibitor according to [1], which is a group represented by the formula: [3] A is the formula:

Embedded image The fat accumulation inhibitor according to [1], which is a group represented by the formula: [4] A is the formula:

Embedded image The fat accumulation inhibitor according to [1], which is a group represented by the formula: [5] The fat accumulation inhibitor according to any one of [1] to [4], which is an antiobesity agent. [6] The fat accumulation inhibitor according to any one of [1] to [4], which is a therapeutic agent for diabetes. [7] The fat accumulation inhibitor according to any of [1] to [4], which is a therapeutic agent for hyperlipidemia.

Various groups in the present invention are described below. Unless otherwise specified, the description of each group includes the case where it is part of another substituent. When the number of carbon atoms is specified in the above [1] for each group, the compound of the present invention is limited to those having the number of carbon atoms. Examples of the alkyl group include methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, neopentyl, and n-hexyl. And straight-chain or branched alkyl groups having 1 to 8 carbon atoms such as n-heptyl group and n-octyl group. Examples of the acyl group include an alkanoyl group and an aroyl group. Examples of the alkanoyl group include a group in which a hydrogen atom or an alkyl group is bonded to one of two bonding hands of a carbonyl group. Examples of the aroyl group include a group in which an aryl group is bonded to one of two bonding hands of a carbonyl group. Examples of the aryl group include groups having 10 or less carbon atoms. Specifically, for example, a phenyl group, a 1-naphthyl group,
-Naphthyl group and the like. As the alkoxy group,
Examples include a group in which an alkyl group is bonded to one of two bonding hands of an oxygen atom. Examples of the alkylthio group include a group in which an alkyl group is bonded to one of two bonding hands of a sulfur atom. Examples of the alkenyl group include a vinyl group, an allyl group, a 1-methylvinyl group, a 1-propenyl group, a 2-butenyl group, a 2-methyl-2-propenyl group, a 2-pentenyl group,
3-methyl-2-butenyl group, 3,3-dimethyl-1-butenyl group, 2-methyl-2-butenyl group, 1-hexenyl group, 2
-Hexenyl group, 5-hexenyl group, 1-heptenyl group,
Examples thereof include a linear or branched alkenyl group having 2 to 8 carbon atoms such as a 1-methyl-1-hexenyl group and a 1-octenyl group.

The alkynyl group includes, for example, ethynyl, propargyl, 1-methylpropargyl, 1-propynyl, 1-butynyl, 3,3-dimethyl-1-butynyl, 1-pentynyl, 1-hexynyl And a straight-chain or branched alkynyl group having 2 to 8 carbon atoms such as a group, 1-heptynyl group and 1-octynyl group. Examples of the alkynylalkenyl group include an alkenyl group substituted by an alkynyl group. For example, 1-butene-3
-Inyl group, 1-penten-3-ynyl group, 2-penten-4-ynyl group, 1-hexene-3-ynyl group, 1-hexene-4-ynyl group, 2-hexene-4-ynyl group, 2 -
Examples thereof include a straight-chain or branched alkynylalkenyl group having 4 to 8 carbon atoms, such as a hexene-5-ynyl group and a 3-hexene-5-ynyl group. As the cycloalkyl group,
For example, carbon atoms such as cyclopropyl group, cyclobutyl group, 2-methylcyclopropyl group, cyclopentyl group, cyclohexyl group, 2-methylcyclohexyl group, 1-methylcyclopropyl group, 4-methylcyclohexyl group, and 4-ethylcyclohexyl group There may be mentioned 3 to 8 cycloalkyl groups.

[0007] Examples of the halogen atom include a fluorine atom, a chlorine atom, a bromine atom and the like. Examples of the haloalkyl group include, for example, a group in which one or more, the same or different, halogen atoms are substituted for the above-mentioned alkyl group having 1 to 8 carbon atoms, and specifically, for example, a difluoromethyl group, a trifluoromethyl group, 2-fluoroethyl group, 2,2-difluoroethyl group, 2,2,2-trifluoroethyl group, 1-fluoroethyl group, 1,2-difluoroethyl group, 3-fluoropropyl group, 1-fluoropropyl group A 2-fluoropropyl group, a 4-fluorobutyl group, a 5-fluoropentyl group, a 6-fluorohexyl group,
7-fluoroheptyl group, 8-fluorooctyl group, 2-
Chloroethyl group, 3-chloropropyl group, 4-chlorobutyl group, 3-bromopropyl group and the like. Examples of the haloalkenyl group include groups in which one or more, the same or different, halogen atoms are substituted for the alkenyl group having 2 to 6 carbon atoms, and specifically, for example, a 1,2-dichlorovinyl group, 2,3-dichloroallyl group, 2
-Iodo-1-fluorovinyl group, 2-chloro-1-fluorovinyl group, 2,2-dichloro-1-fluorovinyl group, 2,
Examples of the haloalkoxy group include a 2-dichlorovinyl group and the like, and a group in which a haloalkyl group is bonded to one of two bonds of an oxygen atom. Examples of the haloalkylthio group include a group in which a haloalkyl group is bonded to one of two bonds of a sulfur atom.

When R ⁵ and R ⁶ are adjacent to each other, a 5-membered or 6-membered ring formed by bonding at both ends (these rings are saturated or unsaturated and have an oxygen atom or a sulfur atom Ring B which is substituted with a halogen atom or an alkyl group having 1 to 4 carbon atoms, which is included in the ring, may be, for example, when ring B is a phenyl group,

Embedded image And the group represented by When the ring B is a pyridine ring or a cyclohexane ring, those substituted with the same ring can be mentioned. When R ⁸ and R ⁹ are adjacent to each other, a saturated 5-membered ring or a saturated 6-membered ring formed by bonding at both ends (these rings are substituted with an alkyl group having 1 to 4 carbon atoms) A) substituted with, for example, when A is pyrazol-5-yl or a substituted group thereof,

Embedded image And the group represented by When the position of a bond is different, and when A is a ring other than a pyrazole ring, those substituted with the same ring can be mentioned. Examples of the substituent of the optionally substituted phenyl group include, for example, a halogen atom, an alkyl group, a haloalkyl group, an alkoxy group or a haloalkoxy group, and one or more, the same or different, may be substituted, Specific examples of such an optionally substituted phenyl group include a phenyl group, a 2-fluorophenyl group, a 3-fluorophenyl group, a 4-fluorophenyl group, a 3,5-difluorophenyl group, and a 4-methoxyphenyl. Group, 4-tolyl group, 4-ethylphenyl group, 3-chlorophenyl group, 4-chlorophenyl group, 4-n-propylphenyl group, 4-isopropylphenyl group, 4- (1,1,2,2-tetrafluoro Ethoxy) phenyl group, 4-trifluoromethylphenyl group,
Examples thereof include a 4-difluoromethoxyphenyl group and a 2,4-dichlorophenyl group.

The "prodrug" includes one that is easily hydrolyzed in a living body to regenerate the compound represented by the general formula (1). Examples of the compound include an alkoxycarbonyl group, an alkylthiocarbonyl group, and an alkylaminocarbonyl group. Further, for example, a compound having an amino group, a compound in which the amino group is substituted with an alkanoyl group to form an alkanoylamino group, a compound in which the amino group is substituted with an alkoxycarbonyl group to form an alkoxycarbonylamino group, and an acyloxymethylamino group. Or a compound that has become hydroxylamine. For example, if the compound has a hydroxyl group,
Examples of the compound include a compound in which the hydroxyl group is substituted with an acyl group to form an acyloxy group, a compound in which a phosphate ester is formed, and a compound in which the hydroxyl group is converted to an acyloxymethyloxy group. The alkyl portion of the group used for these prodrugs may be substituted, for example, by an alkoxy group having 1 to 6 carbon atoms. Preferred examples include, for example, a compound in which a carboxyl group is an alkoxycarbonyl group, for example, a lower (for example, 1 to 6 carbon atoms) alkoxycarbonyl group such as methoxycarbonyl or ethoxycarbonyl, or methoxymethoxycarbonyl or ethoxymethoxycarbonyl. And lower (e.g., 1 to 6 carbon atoms) alkoxycarbonyl groups substituted by an alkoxy group such as, 2-methoxyethoxycarbonyl, 2-methoxyethoxymethoxycarbonyl or pivaloyloxymethoxycarbonyl.

The compound represented by the general formula (1) is disclosed in, for example, JP-A-3-223256 and JP-A-3-81.
It can be manufactured by the method described in Japanese Patent Publication No. 266 or the like. Pharmaceutically acceptable salts of the compound represented by the general formula (1) or a prodrug thereof include pharmaceutically acceptable acid addition salts and base addition salts. Examples of the acid addition salts include inorganic acid addition salts and organic acid addition salts. Examples of the inorganic acid addition salts include hydrochloride, hydrogen bromide, sulfate, and phosphate, and examples of the organic acid addition salts include acetate. Methanesulfonate, benzenesulfonate, citrate, fumarate, maleate, succinate, tartrate, malate and the like. Base addition salts include inorganic base addition salts and organic base addition salts,
Examples of the inorganic base addition salt include alkali metal salts such as sodium salt and potassium salt, alkaline earth metal salts such as magnesium salt and calcium salt, and ammonium salt. Examples of the organic base addition salt include arginine salt and lysine salt And the like. Such salts can be easily produced by known means. The compounds of the present invention also include solvates such as hydrates thereof. The compound of the present invention may have optical isomers derived from, for example, R ² and R ³ , and the compound of the present invention includes any of these isomers and a mixture of these isomers. , Each of which can be used as a medicament.

The compound of the present invention has an action of suppressing fat accumulation in adipocytes, and can be used for prevention and treatment of adipose tissue enlargement and associated diseases. More specifically, it is useful as a prophylactic and / or therapeutic agent for obesity, glucose intolerance, diabetes, hyperglyceridemia, hyperlipidemia, etc. associated with an increase in visceral adipose tissue and body fat, and It is expected to be effective for the prevention and / or treatment of hypertension, coronary artery disease, obstructive arteriosclerosis and the like. The pharmaceutical composition containing the compound of the present invention can be administered by a usual administration route, for example, oral, intramuscular, intravenous, subcutaneous, intraperitoneal, intranasal or the like. Dosage and frequency vary depending on the route of administration, degree of symptoms, age, body weight, etc.
Although not particularly limited, usually about 1 mg to 1 g per day for an adult is taken once or more times a day (for example, 2 to 1 g).
４4 times). Examples of dosage forms include powders, fine granules, granules, tablets, capsules, suppositories, injections, nasal preparations and the like. At the time of formulation, it is manufactured by an ordinary method using a usual formulation carrier. That is, when preparing an oral preparation, a binder, a disintegrant, a lubricant, a colorant, and the like are added as necessary, and then a tablet, a granule, a powder, a capsule, and the like are made in a conventional manner. When preparing an injection, a pH adjuster, a buffer, a stabilizer, a solubilizing agent, and the like are added as necessary, and an injection is prepared by a conventional method.

Specific examples of the compound of the present invention include the compounds shown in Tables 1 to 40 below. However, these compounds are for illustration only, and the present invention is not limited to these.

Embedded image [Wherein the numbers 2 to 6 represent numbers for indicating the substitution positions. ]

[0013]

[Table 1]

[0014]

[Table 2]

[0015]

[Table 3]

[0016]

[Table 4]

[0017]

Embedded image [Wherein the numbers 2 to 6 represent numbers for indicating the substitution positions. ]

[0018]

[Table 5]

[0019]

Embedded image [Wherein the numbers 2 to 6 represent numbers for indicating the substitution positions. ]

[0020]

[Table 6]

[0021]

Embedded image [Wherein the numbers 2 to 6 represent numbers for indicating the substitution positions. ]

[0022]

[Table 7]

[0023]

Embedded image [Wherein the numbers 2 to 6 represent numbers for indicating the substitution positions. ]

[0024]

[Table 8]

[0025]

[Table 9]

[0026]

[Table 10]

[0027]

[Table 11]

[0028]

Embedded image [Wherein the numbers 2 to 6 represent numbers for indicating the substitution positions. ]

[0029]

[Table 12]

[0030]

Embedded image [Wherein the numbers 2 to 6 represent numbers for indicating the substitution positions. ]

[0031]

[Table 13]

[0032]

Embedded image [Wherein the numbers 2 to 6 represent numbers for indicating the substitution positions. ]

[0033]

[Table 14]

[0034]

Embedded image [Wherein the numbers 2 to 6 represent numbers for indicating the substitution positions. ]

[0035]

[Table 15]

[0036]

Embedded image [Wherein the numbers 2 to 6 represent numbers for indicating the substitution positions. ]

[0037]

[Table 16]

[0038]

Embedded image [Wherein the numbers 2 to 6 represent numbers for indicating the substitution positions. ]

[0039]

[Table 17]

[0040]

Embedded image [Wherein the numbers 2 to 6 represent numbers for indicating the substitution positions. ]

[0041]

[Table 18]

[0042]

Embedded image

[0043]

[Table 19]

[0044]

Embedded image

[0045]

[Table 20]

[0046]

Embedded image [Wherein the numbers 2 to 6 represent numbers for indicating the substitution positions. ]

[0047]

[Table 21]

[0048]

Embedded image [Wherein the numbers 2 to 6 represent numbers for indicating the substitution positions. ]

[0049]

[Table 22]

[0050]

[Table 23]

[0051]

[Table 24]

[0052]

Embedded image [Wherein the numbers 2 to 6 represent numbers for indicating the substitution positions. ]

[0053]

[Table 25]

[0054]

Embedded image [Wherein the numbers 2 to 6 represent numbers for indicating the substitution positions. ]

[0055]

[Table 26]

[0056]

Embedded image [Wherein the numbers 2 to 6 represent numbers for indicating the substitution positions. ]

[0057]

[Table 27]

[0058]

Embedded image [Wherein the numbers 2 to 6 represent numbers for indicating the substitution positions. ]

[0059]

[Table 28]

[0060]

Embedded image [Wherein the numbers 2 to 6 represent numbers for indicating the substitution positions. ]

[0061]

[Table 29]

[0062]

Embedded image [Wherein the numbers 2 to 6 represent numbers for indicating the substitution positions. ]

[0063]

[Table 30]

[0064]

Embedded image [Wherein the numbers 2 to 6 represent numbers for indicating the substitution positions. ]

[0065]

[Table 31]

[0066]

Embedded image [Wherein the numbers 2 to 6 represent numbers for indicating the substitution positions. ]

[0067]

[Table 32]

[0068]

Embedded image [Wherein the numbers 2 to 6 represent numbers for indicating the substitution positions. ]

[0069]

[Table 33]

[0070]

Embedded image [Wherein the numbers 2 to 6 represent numbers for indicating the substitution positions. ]

[0071]

[Table 34]

[0072]

Embedded image [Wherein the numbers 2 to 6 represent numbers for indicating the substitution positions. ]

[0073]

[Table 35]

[0074]

Embedded image [Wherein the numbers 2 to 6 represent numbers for indicating the substitution positions. ]

[0075]

[Table 36]

[0076]

[Table 37]

[0077]

Embedded image [Wherein the numbers 2 to 6 represent numbers for indicating the substitution positions. ]

[0078]

[Table 38]

[0079]

Embedded image [Wherein the numbers 2 to 6 represent numbers for indicating the substitution positions. ]

[0080]

[Table 39]

[0081]

Embedded image [Wherein the numbers 2 to 6 represent numbers for indicating the substitution positions. ]

[0082]

[Table 40]

[0083]

EXAMPLES Next, test examples show that the compounds of the present invention are useful as fat accumulation inhibitors, but the present invention is not limited thereto. In addition, the compound used is shown by the compound number of said Table 1-Table 40.

Test Example 1 (1) Removal of Adipose Tissue 32 14-week-old Wistar male rats (Japan SLC, Inc.) were sacrificed and laparotomized, and adipose tissue attached to mesentery (hereinafter referred to as mesentery) All adipose tissue) was removed. Further, subcutaneous adipose tissue from the abdomen to the thigh (hereinafter referred to as abdominal adipose tissue) was excised from only one side per animal. These excised tissues were each added to a phosphate buffer (0.20 g / L KCl, 0.20 g / L).
KH ₂ PO ₄ , 8.00 g / L NaCl, 2.16 g
_{/ L Na 2 HPO 4 · 7H} 2 O, 100 units / ml
Penicillin (GIBCO), 100 μg / ml streptomycin (GIBCO), 250 ng / ml
Amphotericin (GIBCO)) and washed at room temperature.

(2) Preparation and Culture of Cells from Test Tissue The following treatment was performed on each of the washed mesenteric adipose tissue and the abdominal adipose tissue. That is, first, these tissues were treated with collagenase (type II or VIII; manufactured by Sigma), penicillin (manufactured by GIBCO), streptomycin (manufactured by GIBCO) and amphotericin (manufactured by GIBCO) at a final concentration of 1 mg / ml, respectively. 1
00 units / ml, 100 μg / ml and 250 n
g / ml of Dulbecco's modified Eagle medium (4.5 g / L D-glucose and 584 mg).
/ LL Contains L-glutamine. GIBCO) 300
The mixture was cut into approximately 5 mm squares using scissors in the ml. Then, it is shaken at 37 ° C. for 60 minutes (about 170 rpm).
And a nylon mesh (80S [mesh size is 250μ
m]. Then, the filtrate (cell suspension) was recovered. After the filtrate was centrifuged at 1,800 rpm for 5 minutes at room temperature, the liquid layer was gently removed by decantation to obtain a sediment. This sedimentation was performed using fetal bovine serum (hereinafter referred to as FB).
Notated as S. ) (GIBCO), ascorbic acid (Wako Pure Chemical Industries, Ltd.), penicillin, streptomycin (GIBCO) and amphotericin (GIBC)
O company) at final concentrations of 10%, 200 μM, 100
Units / ml, 100 μg / ml and 250 ng /
ml of Dulbecco's modified Eagle's medium (4.5 g / L D-glucose and 584 mg / L).
Contains L-glutamine. The suspension was filtered with a nylon mesh (420S [mesh size 25 μm], manufactured by Sanshin Kogyo Co., Ltd.). The filtrate was collected, centrifuged at 1800 rpm at room temperature for 5 minutes, the liquid layer was gently removed by decantation, and the precipitate was suspended again in 50 ml of the above medium (hereinafter referred to as FBS-containing medium). The suspension is centrifuged, the liquid layer is removed, F
The operation of suspending in a BS-containing medium was performed twice more in the same manner as described above. However, at the end, settle 120 ml of FBS
The cells were suspended in the containing medium to prepare a cell suspension. Each 30 ml of the cell suspension was dispensed into a cell culture flask (T150 for adherent cells; manufactured by Iwaki Glass Co., Ltd.) and cultured at 37 ° C. in the presence of 5% CO ₂ . After 2-3 hours from the start of the culture, the medium is removed,
The wall of the flask was washed with 15 ml of the phosphate buffer. After the washing solution was removed and the washing operation was performed again, the phosphate buffer solution was removed, 30 ml of an FBS-containing medium was added to the flask, and the mixture was cultured at 37 ° C. in the presence of 5% CO ₂ . One day after the start of the culture, the medium was removed, and the wall of the flask was washed once with 15 ml of a phosphate buffer.
Ethylenedinitrotetraacetic acid (hereinafter referred to as EDTA) solution (0.05% trypsin, 0.53 mM EDTA.
4Na. GIBCO) to the extent that the cells are immersed,
It was left at 37 ° C. for 5 minutes. To this, an FBS-containing medium was added in an amount about 10 times that of the trypsin-EDTA solution to obtain a cell suspension.

(3) Test of Fat Accumulation Inhibiting Effect A Using cells derived from mesenteric adipose tissue, the compound was examined for its effect of inhibiting fat accumulation. The number of cells in the suspension prepared from the mesenteric adipose tissue in the above (2) was measured using a hemocytometer, and an FBS-containing medium was added so as to be 1.4 × 10 ⁵ cells / ml. Diluted. The diluted solution was dispensed into a 96-well plate (for culturing adherent cells, manufactured by Iwaki Glass Co., Ltd.) in an amount of 100 μl per well, and 37 ° C. in the presence of 5% CO _2.
And cultured. After 2-3 days, the medium is removed from each well of the 96-well plate, and 10 μg / ml insulin (manufactured by Sigma), 0.25 μM dexamethasone [Wako Pure Chemical Industries, Ltd.], 0.5 mM 3-isobutyl-1-methyl −
100 μl of an FBS-containing medium containing xanthine (manufactured by Sigma) and 5 μM 15-deoxy-Δ ^12,14 -prostaglandin J ₂ (manufactured by Cayman) was added to each well, and the mixture was incubated at 37 ° C. in the presence of 5% CO ₂ For 2 days. Then, the medium was removed from each well, 10 [mu] g / ml insulin and 5 [mu] M 15-deoxy - [delta ^{12, 14} - and the FBS-containing medium 100μl containing prostaglandin J ₂ were added to each well. For another 2 days in the presence of 5% CO ₂
After culturing at 37 ° C., the medium in each well was removed and 10 μl
g / ml insulin, 5 μM 15-deoxy-Δ
^12,14 -prostaglandin J ₂ , test compound 50 μM,
FBS-containing medium 1 containing 0.5% dimethyl sulfoxide (hereinafter referred to as DMSO) [Wako Pure Chemical Industries, Ltd.]
00 μl was added to each well and cultured similarly. In addition,
10 μg / ml insulin was added to the test compound-free group,
FBS-containing medium 100 containing 5 μM 15-deoxy-Δ ^12,14 -prostaglandin J ₂ and 0.5% DMSO
μl was added, and the cells were cultured in the same manner.

After culturing for 2 days, oil red O [S
udan II, Wako Pure Chemical Industries, Ltd.], and intracellular fat was stained, and the fat content was measured by a colorimetric method.
That is, first, directly into each well containing the cell culture solution,
30 μl of 0.075% oil red O staining solution / 60% triethyl phosphate solution was added. After leaving at room temperature for 30 minutes, the medium containing Oil Red O staining solution was removed, and 20%
100 μl of an aqueous triethyl phosphate solution was added to each well. After the addition, the 20% aqueous triethyl phosphate solution was removed from each well, and 100 μl of the same aqueous solution was newly added to each well. After repeating the above operation, the cell lysate (2
100 μl (% SDS, 0.2 N NaOH) was added to each well. After keeping the temperature at 37 ° C. for 3 hours or longer, 490 was measured using a plate reader (Vmax, manufactured by Beckman).
The absorbance at a wavelength of nm was measured (hereinafter referred to as measured value 1). In the test compound-free group, intracellular fat was stained in the same manner, and the absorbance was measured (hereinafter referred to as measured value 2). From these measured values, the fat accumulation inhibition rate by the test compound was calculated according to the following formula. Fat accumulation inhibition rate (%) = {(measured value 2−measured value 1) / measured value 2} × 100 The effect criterion is: A: fat accumulation inhibition rate is 50% or more B: fat accumulation inhibition rate is 10% or more 50 % C: Fat accumulation suppression rate was less than 10%. Table 41 shows the results.

[0088]

[Table 41]

(4) Test B on Fat Accumulation Suppressing Effect The fat accumulation suppressing effect of the compound was examined using cells derived from abdominal adipose tissue. The number of cells in the suspension prepared from the abdominal adipose tissue in the above (2) was measured using a hemocytometer, and an FBS-containing medium was added at 1.4 × 10 ⁵ cells / ml. Diluted. Using the diluent,
Following the same operation as in (3), cell culture is performed, the test compound is added, and the fat of the cells to which the test compound has been added and the fat of the cells to which the test compound has not been added are oil red O [Sudan II]. , Wako Pure Chemical Industries, Ltd.], and their absorbance was measured (hereinafter, the measured value of the cells to which the test compound was added was measured value 3, and the measurement of the cells to which the test compound was not added). The value is referred to as measured value 4.)
From these measured values, the fat accumulation inhibition rate by the test compound was calculated according to the following formula. Fat accumulation inhibition rate (%) = {(measured value−measured value 3) / measured value 4} × 100 The effect criterion is A: fat accumulation inhibition rate of 50% or more B: fat accumulation inhibition rate of 10% or more % C: Fat accumulation suppression rate was less than 10%. The results are shown in Table 42.

[0090]

[Table 42]

Test Example 2 (1) Preparation of Adipose Tissue Piece In the same manner as in Test Example 1, mesenteric adipose tissue and abdominal adipose tissue were excised from 14-week-old male Wistar rats.
The following processing was performed for each of them. That is, first, the extirpated tissue was mixed with 2 ml of Dulbecco's modified Eagle medium (4.5 g / LD-glucose and 584 mg).
/ LL Contains L-glutamine. After washing with GIBCO), the pieces were cut into about 1 mm squares using scissors in the same medium.

(2) Test of Fat Accumulation Inhibiting Effect C Using the mesenteric adipose tissue or the abdominal adipose tissue prepared in (1), the compound was examined for the effect of inhibiting fat accumulation. That is, first, Dulbecco's modified Eagle's medium-low glucose (containing 1.0 g / L D-glucose and 584 mg / L L-glutamine is contained in a 48-well plate (for adherent cell culture; manufactured by Sumitomo Bakelite Co., Ltd., GIBCO).
500 μl per well was dispensed.
The test compound dissolved in MSO was added so that the final concentration of the test compound was 50 μM and the final concentration of DMSO was 0.5%. In each well, the above-mentioned adipose tissue piece 50-100
mg, and cultured at 37 ° C. for 30 minutes in the presence of 5% CO ₂ . In the test compound-free group, DMSO alone was used instead of the above-mentioned DMSO solution of the test compound, and the final concentration of DMSO alone was 0.1.
5%, and a fat tissue piece was added in the same manner as described above, followed by culturing in the same manner. 30 minutes after the start of the culture, a radioisotope-labeled glucose solution (D- [U- ¹⁴ C]
glucose. 7.4 MBq / ml. Amersham) 1
5 μl was added to each well and cultured at 37 ° C. for 7 hours in the presence of 5% CO ₂ .

After the culture, the tissue pieces in each well were treated with heptane / isopropanol (heptane: isopropanol = 3:
2) Transferred to 750 μl and left at room temperature for about 15 hours.
Then, the tissue pieces were removed from the above solution, and heptane and isopropanol were volatilized.
was dissolved in 120 μl of chloroform / methanol (chloroform: methanol = 2: 1).
μl was transferred to a thin-layer chromatography plate (K5 silica gel 150 Å; manufactured by Whatman.
Indicated as TLC plate. ) Spotted on. Hexane: ethyl ether: acetic acid (75: 25: 1) in a closed container
After the developing solvent is added and the TLC plate is developed,
The TLC plate was dried at room temperature, and the plate was exposed to an imaging plate (BAS3-2040, manufactured by Fuji Film Co., Ltd.) for 4 to 5 hours. The exposed imaging plate was analyzed with an image analyzer (BAS2000 BioImage Analyzer, manufactured by Fuji Film Co., Ltd.), and the radioactivity of the portion corresponding to the developed position of triglyceride on the TLC plate was measured (hereinafter referred to as measured value 5). .). The test compound-free group was treated in the same manner and subjected to thin-layer chromatography to measure the radioactivity of the portion corresponding to the development position of triglyceride on the TLC plate (hereinafter referred to as measured value 6). From these measured values, the inhibition ratio of fat (triglyceride) accumulation per unit volume of adipose tissue by the test compound was calculated according to the following formula. Fat accumulation inhibition rate (%) = {(measured value−measured value 5) / measured value 6} × 100

In the test using mesenteric adipose tissue, the inhibitory rate of fat accumulation by the test compound was as follows.
The effect criteria were as follows: A: fat accumulation inhibition rate of 40% or more B: fat accumulation inhibition rate of 10% or more and less than 40% C: fat accumulation inhibition rate of less than 10%. The results are shown in Table 43.

[0095]

[Table 43]

In the test using the abdominal adipose tissue, the test compound exhibited the following fat accumulation inhibition rates.
The effect criteria were as follows: A: fat accumulation inhibition rate of 40% or more B: fat accumulation inhibition rate of 10% or more and less than 40% C: fat accumulation inhibition rate of less than 10%. The results are shown in Table 44.

[0097]

[Table 44]

[0098]

Industrial Applicability According to the present invention, it is possible to provide a fat accumulation inhibitor containing a compound represented by the general formula (1), a prodrug thereof, or a pharmaceutically acceptable salt thereof. Furthermore, by suppressing fat accumulation in adipose tissue using the fat accumulation inhibitor of the present invention, obesity, diabetes, hyperlipidemia, etc. associated with an increase in body fat and adipose tissue in the abdominal cavity. The disease can be prevented and treated.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat ゛ (Reference) A61K 31/433 A61K 31/433 A61P 3/04 A61P 3/04 3/06 3/06 3/10 3 / 10 // C07D 231/14 C07D 231/14 231/56 231/56 C 263/34 263/34 277/56 277/56 285/06 285/06 401/12 401/12 405/12 405/12 409 / 12 409/12 413/12 413/12 (72) Inventor Yuko Murakami 4-2-1 Takashi, Takarazuka-shi, Hyogo F-term in Sumitomo Kagaku Kogyo Co., Ltd. 4C033 AD03 AD20 4C036 AD04 AD23 AD30 4C056 AA01 AB01 AC02 AD01 AE03 BA03 BB14 BC01 4C063 AA01 BB07 CC22 CC76 CC81 CC82 CC94 DD12 DD22 DD52 EE01 4C086 AA01 AA02 BC36 BC37 BC69 BC82 BC85 GA02 GA03 GA07 GA08 MA01 MA04 NA14 ZA70 ZC33 ZC35

Claims (7)

[Claims] 1. General formula (1):[Wherein, R¹Is a hydrogen atom, an alkyl group having 1 to 4 carbon atoms
Alternatively, it represents an alkanoyl group having 1 to 8 carbon atoms. R²
Is a hydrogen atom, an alkyl group having 1 to 4 carbon atoms or a cyano group
Represents R³Is a hydrogen atom or an alkyl having 1 to 4 carbon atoms
Represents a group. R⁴Are independent of each other if there are multiple
A hydrogen atom, a halogen atom or an alkyl having 1 to 4 carbon atoms.
Represents a kill group. R⁵Is hydrogen atom, halogen atom, hydroxyl
Group, amino group, cycloalkyl group having 3 to 8 carbon atoms, carbon
Aralkyl having 6 to 10 aryl groups and 2 to 8 carbon atoms
Disubstituted alkyl groups, alkylthio groups having 2 to 8 carbon atoms
Substituted alkyl group, haloalkylthio having 2 to 8 carbon atoms
A substituted alkyl group having 1 to 8 carbon atoms: -NHR⁵¹Represented by
Group having the total carbon number of 2 to 16: -NR⁵²R⁵³so
A group represented by the formula having 1 to 8 carbon atoms: -NHCOR⁵⁴In table
Group represented by the formula having 1 to 8 carbon atoms: -N = CR⁵⁴R^{5 5}so
A group represented by the formula having a total of 2 to 8 carbon atoms: -N = CHNHR
⁵²A group having a total number of carbon atoms of 3 to 8: -N = C
(CH₃) NHR⁵²A group represented by the formula, wherein the total number of carbon atoms is 3 to 1
Formula of 6: -N = CHNR⁵²R⁵³Group represented by
Formula with a prime number of 4 to 16: -N = C (CH₃) NR⁵²R
⁵³Or a formula having 1 to 8 carbon atoms: —NHS
O₂R⁵¹Or a group represented by the formula: -R⁵⁶, -OR
⁵⁶, -SR⁵⁶, -SOR⁵¹, -SO₂R⁵¹if
Ku-OSO₂R⁵¹A group represented by the formula (R⁵¹Is 1 carbon
Alkyl group or haloalkyl group having 1 to 8 carbon atoms
And R⁵²And R⁵³Are each independently carbon
Represents an alkyl group of Formulas 1 to 8,⁵⁴And R⁵⁵Is
Each independently represents a hydrogen atom or an alkyl having 1 to 8 carbon atoms;
Represents a kill group;⁵⁶Is an alkyl group having 1 to 8 carbon atoms,
C1-8 haloalkyl group, C2-8 alk
Nil group, haloalkenyl group having 2 to 8 carbon atoms, 2 to 2 carbon atoms
An alkynyl group of 8 or an alkynyl group having 4 to 8 carbon atoms
Represents a phenyl group). R⁶Is more than one
Hydrogen atom, halogen atom
, Cycloalkyl group having 3 to 6 carbon atoms or total carbon number
Is an alkoxy-substituted alkyl group having 2 to 3 or a group represented by the formula: —R
⁶¹, -OR⁶¹, -SR⁶¹, -SOR⁶², -SO
₂R⁶², -OSO₂R⁶²A group represented by the formula:
⁶¹Is an alkyl group having 1 to 3 carbon atoms, and a halo having 1 to 3 carbon atoms.
Alkyl group, alkenyl group having 2 to 6 carbon atoms, 2 to 2 carbon atoms
6 haloalkenyl groups or alkynyl having 2 to 6 carbon atoms
A group represented by R⁶²Is an alkyl group having 1 to 3 carbon atoms or carbon number
Represents 1 to 3 haloalkyl groups). Also, R⁵When
R⁶Are adjacent to each other, 5
May form a six-membered ring or a six-membered ring, and these rings are
Saturated or unsaturated, oxygen or sulfur atoms
2 to 2 halogen atoms or 1 to 4 carbon atoms
May be substituted with an alkyl group. m is 1 or
Represents an integer of 2. n represents an integer of 1 to 4. A is the formula:Represents a group represented by R⁷, R¹⁰, R¹¹And R
¹²Are each independently a hydrogen atom, an alkyl group having 1 to 4 carbon atoms.
Represents a alkyl group or a haloalkyl group having 1 to 4 carbon atoms.
R⁸And R⁹Are each independently a hydrogen atom or
A halogen atom or a formula: —R⁸¹Or -OR⁸¹
(R⁸¹Is an alkyl group having 1 to 6 carbon atoms,
A haloalkyl group, a cycloalkyl group having 3 to 6 carbon atoms or
Represents an optionally substituted phenyl group)
Represents a group. Also, R⁸And R⁹When both are adjacent, both ends
To form a saturated 5-membered ring or a saturated 6-membered ring
May form a ring, and these rings have 1 to 4 carbon atoms.
It may be substituted with an alkyl group. X and Z are
Each independently represents an oxygen atom or a sulfur atom. Y is
An oxygen atom or a sulfur atom, or a formula: -SO-, -S
O₂-, -CH ₂-, -OCH₂-Or -NR¹³
− (R¹³Represents a hydrogen atom or an alkyl group).
Represents a group. Ring B is a benzene ring, a pyridine ring or
Represents a cyclohexane ring. Or a compound represented by
Prodrugs or their pharmaceutically acceptable salts
A fat accumulation inhibitor containing. 2. A is a compound of the formula: The fat accumulation inhibitor according to claim 1, which is a group represented by the formula: 3. A is a compound of the formula: The fat accumulation inhibitor according to claim 1, which is a group represented by the formula: 4. A is a compound of the formula: The fat accumulation inhibitor according to claim 1, which is a group represented by the formula: 5. The fat accumulation inhibitor according to any one of claims 1 to 4, which is an antiobesity agent. 6. The fat accumulation inhibitor according to claim 1, which is a therapeutic agent for diabetes. 7. The fat accumulation inhibitor according to claim 1, which is a therapeutic agent for hyperlipidemia.