JP3274011B2 - Cyclopentaquinoline derivative - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to a novel cyclopentaquinoline derivative. More specifically, the present invention has 9-amino-2,3,5,6,7,8-hexahydro which has an acetylcholinesterase inhibitory activity shown in JP-B-3-54922 and has a therapeutic effect on Alzheimer's disease. −
1H-cyclopenta (b) quinoline [compound (2)]

Embedded image The present invention relates to a cyclopentaquinoline derivative which is useful as a hapten when measuring the concentration in a biological fluid by an enzyme immunoassay.

[0002]

2. Description of the Related Art Conventionally, immunological methods have been generally used as a method for measuring the concentration of a drug or the like in a biological fluid. However, there is no example of measuring the compound (2) by an immunological method to date.

[0003]

Generally, compound (2)
Are called haptens and have antigenicity but no immunogenicity. Therefore, in order to produce an antibody for these haptens, an animal must be immunized after covalently bonding an amino group, a carboxyl group, etc. in the hapten to an immunogenic carrier such as a protein. However, when a protein is directly covalently bonded to the amino group of the compound (2), the amino group of the compound (2) is poor in reactivity, so that it is difficult to obtain a covalent bond with the protein, and the obtained covalent bond is The antibody was not used to immunize the animal, and a favorable antibody was not obtained. Therefore, development of a hapten for preparing an immunogen required for immunological measurement of compound (2) has been desired.

[0004]

Means for Solving the Problems The present inventors have carried out various studies on a method for immunologically measuring compound (2), particularly on a hapten required for producing an immunogen, and as a result, compound (2) It has been found that a compound in which a carboxypropyl group or the like is bound to the 5-position is a preferred hapten for preparing an immunogen used for measuring compound (2) by enzyme immunoassay (EIA). The present invention has been completed by further research based on such findings.

That is, the present invention provides the following formula (1)

[0006]

Embedded image

(Wherein, n represents an integer of 0 to 5). It is about.

Hereinafter, the present invention will be described in detail. Equation (1)
Is a novel compound and has the following reaction formula

[0009]

Embedded image

(Wherein n represents an integer of 0 to 5, R represents an alkyl group and X represents a halogen atom.) 1-Pyrrolidino-1-cyclohexene (6) and a compound Compound (8) obtained by reacting (7) in an organic solvent is reacted with water by adding water to produce cyclohexanone derivative (3), and then cyclohexanone derivative (3) and 2-amino-1-cyclopentene- 1-carbonitrile (4) is converted to ethyl polyphosphate (PP
It can be produced by reacting in the presence of E) and hydrolyzing the obtained compound (5). (JP-A-63-297
367) Among the compounds (1), compounds wherein n is 1 to 4 are particularly preferred in the present invention.

[0011]

Examples of the present invention will be described below. Example 1 9-amino-2,3,5,6,7,8-hex
Sahydro-1H-cyclopenta (b) quinoline-5-ca
Synthesis of ruboxylic acid i) Synthesis of 9-amino-2,3,5,6,7,8-hexahydro-1H-cyclopenta (b) quinoline-5-carboxylic acid ethyl ester 2-amino-1
-Cyclopentene-1-carbonitrile 18.36 g (0.17
M) and 27.2 g of 2-ethoxycarbonylcyclohexanone
(0.16 M) was reacted with ethyl polyphosphate in 350 ml of dry chloroform by heating and stirring at 37 ° C. for 18 hours. This was subjected to silica gel column chromatography (chloroform /
Purified with methanol / concentrated aqueous ammonia = 14/1 / 0.1)
1.5 g (3.4% yield) of crystals were obtained. Melting point: 133-135 ° C

Ii) 9-amino-2,3,5,6,7,8
-Hexahydro-1H-cyclopenta (b) quinoline-
Synthesis of 5-Carboxylic Acid 0.3 g (12 mM) of the above product was dissolved in 30 ml of methanol.
3 ml of N sodium hydroxide was added and stirred for 3 hours. After adding 1.5 ml of 2N hydrochloric acid, the solvent was distilled off, and the obtained residue was recrystallized from methanol and ether to give 0.25 g (yield 93.3 g).
%) Of the compound was obtained. IR: νKBrmaxcm ^-1 3230 (NH) 1720 (C = O) ¹ H-NMR: DMSO-d ₆ , PPM 1.46 to 2.55 (9H, m) 2.75 (2H, t) 2.98 (2H, m) ¹³ C-NMR : D ₂ O, DDS, PPM 178 (C = O)

Embodiment 2:2- [9-amino-2,3,
5,6,7,8-hexahydro-1H-cyclopenta
(B) quinolin-5-yl] acetic acid
Success i) 2- [9-amino-2,3,5,6,7,8-hex
Sahydro-1H-cyclopenta (b) quinolin-5-i
[A] Synthesis of Acetic Acid Ethyl Ester 40 g of ethyl polyphosphate dissolved in 100 ml of anhydrous chloroform
And ethyl-2-cyclohexenone-acetate
5.63 g (30.6 mM) and 2-amino-1-cyclopente
3.25 g (30.1 mM) of 1-carbonitrile was added and the mixture was heated at 40 ° C.
For 20 hours. After cooling, add 28% ammonia with ice
A.Drip into water to make it alkaline, and add
Extracted times. After washing the chloroform layer with water, anhydrous sodium sulfate
And dried under reduced pressure. The residue is ethyl acetate
After recrystallizing with ethyl acetate, recrystallize with ethyl acetate to give 6.212 g (yield
The yield was 76.5%). Melting point: 117-120 ° C

Ii) 2- [9-amino-2,3,5,6,
Synthesis of 7,8-Hexahydro-1H-cyclopenta (b) quinolin-5-yl] acetic acid 500 mg (1.82 mM) of the above product was treated with 1N sodium hydroxide
2.3 ml was added and the mixture was heated and reacted at 90 ° C. for 4 hours. After cooling,
The mixture was neutralized with 2.3 ml of 1N hydrochloric acid, and the precipitated crystals were collected by filtration and washed with water and ethyl acetate to obtain 407 mg of white powdery crystals. Yield: 407mg (91.1%) IR: νKBrmaxcm -1 3375,3200 (NH) 2650 (NH +) 1670,1625 (C = N, C = C) 1565,1390 (COO -) 1 H over NMR: DMSO- d ₆ , PPM 1.34 to 1.46 (1H, m) 1.55 to 1.69 (1H, m) 1.85 to 1.98 (2H, m) 2.02 to 2.12 (2H, dt) 2.22 to 2.35 (2H, m) 2.40 to 2.50 (1H, m m) 2.58~2.66 (1H, m) 2.69 (2H, t) 2.83 (2H, t) 2.96~3.09 (1H, m) 13 C over _{NMR: DMSO-d 6, PPM} 35.8 (CH) 43.4 (CH 2) 112.9, 117.7, 151.2, 153.5, 156.4 (pyridine ring) 174.3 (COOH) 20.8, 22.1, 23.0, 27.4, 30.3, 32.0 (CH ₂ )

Embodiment 3:4- [9-amino-2,3,
5,6,7,8-hexahydro-1H-cyclopenta
(B) Synthesis of quinolin-5-yl] butyric acid i) Synthesis of ethyl-2-cyclohexenone butyrate 1-pyrrolidino-1-cyclohexene was added to 70 ml of ethanol.
Non 25.0 g, 4-bromobutyric acid ethyl es
33.6 g of Ter was added, and the mixture was refluxed for 24 hours. Product
To dissolve, 60 ml of water was added and the mixture was refluxed for 5 hours. More water 60
ml was added and extracted twice with ether. Ether layer is saturated
After washing with brine, dry over anhydrous sodium sulfate and
Distilled off. The residue was purified by vacuum distillation. Yield: 5.1g (15%) Boiling point: 136-141 ° C / 3mmHg

Ii) 4- [9-amino-2,3,5,6,
Synthesis of 7,8-Hexahydro-1H-cyclopenta (b) quinolin-5-yl] butyric acid ethyl ester 57 g of ethyl polyphosphate was dissolved in 50 ml of anhydrous chloroform, and 5.07 g (23.9 mM) of the above product was added thereto. 2.58 g of amino-1-cyclopentene-1-carbonitrile (2
3.9 mM) and reacted at 55 ° C. for 10 hours. After cooling, the mixture was added dropwise to 28% aqueous ammonia with ice to make it alkaline, and extracted twice with chloroform. The chloroform layer was washed with water, dried over anhydrous sodium sulfate, and distilled off under reduced pressure. The residue was crystallized from ethyl acetate to give 3.28 g of crude crystals. 1.50 g of crude crystals were obtained from the mother liquor by silica gel column chromatography. These were combined to obtain 4.20 g (58.0%) of white powdery crystals from a mixed solution of ethyl acetate-ether. Melting point: 112-114 ° C

Iii) 4- [9-amino-2,3,5,
6,7,8-hexahydro-1H-cyclopenta (b)
Synthesis of quinolin-5-yl] butyric acid 0.1N sodium hydroxide was added to 0.337 g (1.10 mM) of the above product.
13 ml was added, and the mixture was heated and reacted at 90 ° C. for 2 hours. After cooling,
The mixture was acidified with 15 ml of 0.1N hydrochloric acid, a small amount of saturated saline was added, and the mixture was washed with chloroform. Crystals precipitated from the aqueous layer were collected by filtration and washed with a small amount of water. The crude crystals were recrystallized from water to obtain 0.21 g (68.6%) of white powdery crystals. Melting point: 242.0-243.5 ° C (decomposition) IR: νKBrmax cm ⁻¹ 3400, 3350 (NH ₂ ) 3000-2500 (OH) 1710 (C = O) ¹ H-NMR: CDCl ₃ , TMS, PPM 1.51-1.75 (6H, m) 2.05 to 2.12 (2H, m) 2.18 to 2.29 (3H, m) 2.41 to 2.49 (3H, m) 2.68 to 2.72 (2H, t) 2.85 (1H, br) 2.95 to 2.99 (2H, t) 7.43 ( 2H, br,) ¹³ C-NMR: CDCl ₃ , TMS, PPM 174.2 (C = O) 154.3, 153.0 (pyridine ring) 149.0 (pyridine ring) 118.8, 113.6 (pyridine ring) 35.1, 33.5, 33.2, 30.7, 27.6 , 23.8, 22.2, 22.0, 21.8, 16.6
(CH ₂ ) MS: m / Z 246 (M ⁺ )

[0018]

Industrial Applicability The cyclopentaquinoline derivative according to the present invention is covalently bound to a protein to form an immunogen, and then parenterally administered to an animal to thereby obtain a compound having a high affinity for compound (2). Efficiently produce antibodies with high titers. Therefore, the compounds of the present invention are useful for obtaining antigens, antibodies, and the like necessary for establishing an immunoassay method capable of measuring the concentration of compound (2) in a biological fluid with high sensitivity, accuracy, speed, and convenience. is there.

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Claims (2)

(57) [Claims] (1) Formula (1) (Wherein n represents an integer of 0 to 5). 2. The cyclopentaquinoline derivative according to claim 1, wherein n is an integer of 1 to 4.