Substituted ureas and methods of treating mental illness using the same

Abstract

A cyclohexane derivative as shown by formula IB or a stereoisomer or a salt thereof, has a high affinity for D.sub.3 receptors and 5-hydroxytryptamine, has a lower affinity for D.sub.2 receptors, shows a high selectivity for D.sub.3/D.sub.2 receptors, and can be used as a therapeutic drug against neuropsychiatric diseases. ##STR00001##
wherein; X is N or CH; R is ##STR00002##
R is optionally substituted with one or more substituents selected from the group consisting of F, Cl, Br, I and C.sub.1-C.sub.6 alkyl; and the C.sub.1-C.sub.6 alkyl is optionally substituted with one or more substituents selected from the group consisting of F, Cl, Br, and I.

Claims

1. A compound shown by the formula IB: ##STR00038## or a pharmaceutically acceptable salt or stereoisomer thereof, wherein: X is N or CH; and R is ##STR00039## where R is optionally substituted with one or more substituents selected from the group consisting of F, Cl, Br, I and C.sub.1-C.sub.6 alkyl; and further where the C.sub.1-C.sub.6 alkyl is optionally substituted with one or more substituents selected from the group consisting of F, Cl, Br and I.

2. The compound according to claim 1, or a pharmaceutically acceptable salt or stereoisomer thereof, wherein R is substituted with one C.sub.1-C.sub.4 alkyl, where the C.sub.1-C.sub.4 alkyl is substituted with one or more F.

3. The compound according to claim 1, or a pharmaceutically acceptable salt or stereoisomer thereof, wherein the pharmaceutically acceptable salt is an acid addition salt formed with an acid selected from the group consisting of hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, formic acid, acetic acid, oxalic acid, malonic acid, maleic acid, fumaric acid, succinic acid and benzoic acid.

4. The compound according to claim 1, or a pharmaceutically acceptable salt or stereoisomer thereof, wherein the stereoisomer is the cis-stereoisomer or the trans-stereoisomer.

5. The compound according to claim 1, or a pharmaceutically acceptable salt or stereoisomer thereof, wherein the compound is selected from the group consisting of: N-[trans-4-[2-[4-(benzo[b]thiophene)-7-piperazinyl]ethyl]cyclohexyl]-N,N-dimethylurea, compound 1; N-[trans-4-[2-[7-(benzo[b]thiophene)-7-piperazinyl]ethyl]cyclohexyl]-N,N-dimethylurea, compound 2; N-[trans-4-[2-[4-(benzo[c]thiophene)-7-piperazinyl]ethyl]cyclohexyl]-N,N-dimethylurea, compound 3; N-[trans-4-[2-[4-(6-fluorobenzo[d]isoxazol)-3-piperazinyl]ethyl]cyclohexyl]-N,N-dimethylurea, compound 5; N-[trans-4-[2-[4-(3-chlorobenzo[d]isoxazol)-6-piperazinyl]ethyl]cyclohexyl]-N,N-dimethylurea, compound 6; N-[trans-4-[2-[4-(6-fluorobenzo[d]isoxazol)-3-piperidinyl]ethyl]cyclohexyl]-N,N-dimethylurea, compound 7 N-[trans-4-[2-[4-(benzo[b]thiophene)-7-piperidinyl]ethyl]cyclohexyl]-N,N-dimethylurea, compound 8; N-[trans-4-[2-[7-(benzo[b]thiophene)-7-piperidinyl]ethyl]cyclohexyl]-N,N-dimethylurea, compound 9; N-[trans-4-[2-[4-(benzo[c]thiophene)-7-piperidinyl]ethyl]cyclohexyl]-N,N-dimethylurea, compound 10; N-[trans-4-[2-[4-(benzo[d]isothiazolyl)-3-piperidinyl]ethyl]cyclohexyl]-N,N-dimethylurea, compound 11; N-[trans-4-[2-[4-(3-chlorobenzo[d]isoxazol)-6-piperidinyl]ethyl]cyclohexyl]-N,N-dimethylurea, compound 12; N-[trans-4-[2-[4-(3-methylbenzo[d]isoxazol)-6-piperazinyl]ethyl]cyclohexyl]-N,N-dimethylurea, compound 13; and N-[trans-4-[2-[4-(6-methylbenzo[d]isoxazol)-4-piperazinyl]ethyl]cyclohexyl]-N,N-dimethylurea, compound 14.

6. A pharmaceutical composition comprising as an active ingredient a compound according to claim 1, or a pharmaceutically acceptable salt or stereoisomer thereof, and a pharmaceutically acceptable excipient.

7. The pharmaceutical composition according to claim 6, wherein the pharmaceutical composition is a solid or a liquid preparation for oral, gastrointestinal, buccal, sublingual, nasal, rectal or transdermal administration.

8. The pharmaceutical composition according to claim 7, wherein the pharmaceutical composition is a solid tablet.

9. The pharmaceutical composition according to claim 8, wherein the compound in the solid tablet is N-[trans-4-[2-[4-(benzo[b]thiophene)-7-piperazinyl]ethyl]cyclohexyl]-N,N-dimethylurea.

10. The pharmaceutical composition according to claim 7, wherein the pharmaceutical composition is a suspension.

11. The pharmaceutical composition according to claim 10, wherein the compound in the suspension is N-[trans-4-[2-[4-(benzo [b]thiophene)-7-piperazinyl]ethyl]cyclohexyl]-N,N-dimethylurea.

12. A method for inhibiting dopamine D2 receptor binding activity in a patient, comprising administering to a patient in need thereof a therapeutically effective amount of the pharmaceutical composition according to claim 6.

13. The method according to claim 12, wherein the patient suffers from one or more disorders selected from the group consisting of a phobia, a mental disorder, a mood disorder, a cognitive disorder and an obsessive-compulsive disorder.

14. The method according to claim 13, wherein the mental disorder, mood disorder or cognitive disorder is selected from the group consisting of mental confusion, schizophrenia, depression, anxiety, dysphrenia and a bipolar disorder.

15. A method for inhibiting 5-hydroxytryptamine 1A receptor binding activity in a patient, comprising administering to a patient in need thereof a therapeutically effective amount of the pharmaceutical composition according to claim 6.

16. The method according to claim 15, wherein the patient suffers from one or more disorders selected from the group consisting of a phobia, a mental disorder, a mood disorder, a cognitive disorder and an obsessive-compulsive disorder.

17. The method according to claim 16, wherein the mental disorder, mood disorder or cognitive disorder is selected from the group consisting of mental confusion, schizophrenia, depression, anxiety, dysphrenia and a bipolar disorder.

18. A process for preparing a compound according to claim 1 shown by the formula IB: ##STR00040## wherein the process comprises: reacting a compound of the formula II: ##STR00041## wherein: X is N or CH: and ##STR00042## R is where R is optionally substituted with one or more substituents selected from the group consisting of F, Cl, Br, I and C.sub.1-C.sub.6 alkyl; and further where the C.sub.1-C.sub.6 alkyl is optionally substituted with one or more substituents selected from the group consisting of F, Cl, Br and I; with a compound of the formula III: ##STR00043## in the presence of a base selected from the group consisting of triethylamine, diisopropylethylamine, pyridine, sodium carbonate, potassium carbonate, sodium bicarbonate and potassium bicarbonate, to provide the compound according to claim 1 shown by the formula IB above.

19. A process for preparing the pharmaceutical composition according to claim 6, wherein the process comprises admixing a pharmaceutically acceptable excipient with the compound according to claim 1, or a pharmaceutically acceptable salt or stereoisomer thereof.

20. A compound shown by the formula I: ##STR00044## or a pharmaceutically acceptable salt or stereoisomer thereof, wherein: R is ##STR00045## where R is optionally substituted with one or more substituents selected from the group consisting of F, Cl, Br, I and C.sub.1-C.sub.6 alkyl; and further where the C.sub.1-C.sub.6 alkyl is optionally substituted with one or more substituents selected from the group consisting of F, Cl, Br and I.

21. The compound according to claim 20, or a pharmaceutically acceptable salt or stereoisomer thereof, wherein R is substituted with one C.sub.1-C.sub.4 alkyl, where the C.sub.1-C.sub.4 alkyl is substituted with one or more F.

22. The compound according to claim 20, or a pharmaceutically acceptable salt or stereoisomer thereof, wherein the pharmaceutically acceptable salt is an acid addition salt formed with an acid selected from the group consisting of hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, formic acid, acetic acid, oxalic acid, malonic acid, maleic acid, fumaric acid, succinic acid and benzoic acid.

23. The compound according to claim 20, or a pharmaceutically acceptable salt or stereoisomer thereof, wherein the stereoisomer is the trans-stereoisomer.

24. The compound according to claim 20, or a pharmaceutically acceptable salt or stereoisomer thereof, wherein the compound is selected from the group consisting of: N-[trans-4-[2-[4-(benzo[b]thiophene)-7-piperazinyl]ethyl]cyclohexyl]-N,N-dimethylurea, compound 1; N-[trans-4-[2-[7-(benzo[b]thiophene)-7-piperazinyl]ethyl]cyclohexyl]-N,N-dimethylurea, compound 2; N-[trans-4-[2-[4-(benzo[c]thiophene)-7-piperazinyl]ethyl]cyclohexyl]-N,N-dimethylurea, compound 3; N-[trans-4-[2-[4-(6-fluorobenzo[d]isoxazol)-3-piperazinyl]ethyl]cyclohexyl]-N,N-dimethylurea, compound 5; N-[trans-4-[2-[4-(3-chlorobenzo[d]isoxazol)-6-piperazinyl]ethyl]cyclohexyl]-N,N-dimethylurea, compound 6; N-[trans-4-[2-[4-(3-methylbenzo[d]isoxazol)-6-piperazinyl]ethyl]cyclohexyl]-N,N-dimethylurea, compound 13; and N-[trans-4-[2-[4-(6-methylbenzo[d]isoxazol)-4-piperazinyl]ethyl]cyclohexyl]-N,N-dimethylurea, compound 14.

25. A pharmaceutical composition comprising as an active ingredient a compound according to claim 20, or a pharmaceutically acceptable salt or stereoisomer thereof, and a pharmaceutically acceptable excipient.

26. A process for preparing a compound according to claim 20 shown by the formula I: ##STR00046## wherein the process comprises: reacting a compound of the formula II: ##STR00047## wherein: R is ##STR00048## where R is optionally substituted with one or more substituents selected from the group consisting of F, Cl, Br, I and C.sub.1-C.sub.6 alkyl; and further where the C.sub.1-C.sub.6 alkyl is optionally substituted with one or more substituents selected from the group consisting of F, Cl, Br and I; with a compound of the formula III: ##STR00049## in the presence of a base selected from the group consisting of triethylamine, diisopropylethylamine, pyridine, sodium carbonate, potassium carbonate, sodium bicarbonate and potassium bicarbonate, to provide the compound according to claim 20 shown by the formula I above.

27. A process for preparing the pharmaceutical composition according to claim 25, wherein the process comprises admixing a pharmaceutically acceptable excipient with the compound according to claim 20, or a pharmaceutically acceptable salt or stereoisomer thereof.

Description

DETAILED DESCRIPTION OF THE INVENTION

(1) Materials and agents used in the following examples are commercially available, unless otherwise specified.

EXAMPLE 1

Preparation of 1-benzo[b]thiophene-4-piperazine hydrochloride

(2) ##STR00024##

(3) A mixture of 7.20 g of 4-bromobenzo[b]thiophene, 19.9 g of piperazine anhydride, 4.70 g of sodium tert-butoxide, 0.32 g of (R)-(+)-2,2-bis(diphenylphosphino)-1,1-dinaphthalene (BINAP), 0.63 g of tris(dibenzylideneacetone)dipalladium and 150 ml of toluene was refluxed at nitrogen atmosphere for 1 hour. 150 ml of water was added to the mixture, extracted with 100 ml3 of ethyl acetate, washed with water, dried with magnesium sulfate and then evaporated under reduced pressure to remove the solvent (0.01 MPa, 45 C.). The remainder was purified by silica gel column chromatography (methylene chloride:methanol:25% aqueous ammonia=100:10:1) to obtain 4.60 g of 1-benzo[b]thiophen-4-yl-piperazine as a yellow oil. 2 ml of concentrated hydrochloric acid was added to a methanol solution (25 ml) containing 4.6 g of 1-benzo[b]thiophen-4-yl-piperazine, and evaporated under reduced pressure (0.01 MPa, 45 C.) to remove the solvent. Ethyl acetate (50 ml) was added to the remainder to precipitate and crystallize. The resultant was filtered and then dissolved in 15 ml of methanol under reflux. After cooling to room temperature (25 C.), recrystallization was carried out to give the crystallized 1-benzo[b]thiophene-4-yl-piperazine hydrochloride as colorless needles.

EXAMPLE 2

Preparation of trans-4-[2-[4-(benzo[b]thiophen)-7-piperazinyl]ethyl]cyclohexyl-tert-butyl carbamate

(4) ##STR00025##

(5) 2.54 g (10 mmol) of 1-benzo[b]thiophene-4-piperazine hydrochloride prepared in Example 1 and 2.40 g (10 mmol) of trans-2-{1-[4-(N-tert-butoxycarbony)amino]cyclohexyl}-acetaldehyde were dissolved in 120 ml of dichloromethane. 1.40 ml (10 mmol) of triethylamine was added slowly at room temperature (25 C.2 C.), stirred for 10 minutes, and then 3.16 g (14.8 mmol) of sodium triacetoxyborohydride was added gradually. The mixture was stirred at room temperature for reaction for 24 hours. After the reaction was completed, 120 ml of 10% sodium bicarbonate solution was added. The reaction system was directly extracted and separated, the organic phase was dried with anhydrous sodium sulfate, and filtered and evaporated to dryness. The solid was refluxed to dissolve with 15 ml of ethyl acetate, and then cooled to room temperature (25 C.2 C.), crystallized to give 3.70 g of the desired product.

(6) .sup.1H-NMR (CDCl.sub.3) ppm: 7.81 (1H, brs), 7.78 (1H, d, J=5.5 Hz), 7.73 (1H, d, J=8.1 Hz), 7.41 (1H, m), 7.30 (1H, d, 7.6 Hz), 6.94 (1H, d, J=7.6 Hz), 3.54 (1H, m), 3.35-3.23 (8H, m), 2.46 (2H, m), 1.86-1.65 (8H, m), 1.51-1.49 (1H, m), 1.42 (9H, s), 1.37-1.35 (2H, m).

EXAMPLE 3

Preparation of trans-4-[2-[4-(benzo[b]thiophene)-7-piperazinyl]ethyl]cyclohexylamine

(7) ##STR00026##

(8) 4.43 g of trans-4-[2-[4-(benzo[b]thiophen)-7-piperazinyl]ethyl]cyclohexyl-tert-butyl carbamate prepared in Example 2 was placed in a reaction flask under an ice-water bath. 80 ml of a saturated solution of hydrogen chloride in ethyl acetate was added to the reaction flask. The reaction was stirred for 8 hours for de-protection and finally a white precipitate was formed to give 3.42 g of the hydrochloride of the desired product. The resulting solid was added to 50 ml of dichloromethane solution, and then 50 ml of a saturated solution of sodium bicarbonate was added, stirred for 0.5 hour. This mixture was extracted and separated and the organic phase was concentrated (0.01 MPa, 40 C.) to give 3.30 g of the desired product.

(9) .sup.1H-NMR (CDCl.sub.3) ppm: 7.78 (1H, d, J=5.5 Hz), 7.76 (1H, d, J=8.1 Hz), 7.37 (1H, m), 7.29 (1H, d, 7.6 Hz), 6.96 (1H, d, J=7.6 Hz), 3.48-3.38 (8H, m), 2.53 (1H, m), 2.46 (2H, m), 1.78-1.63 (8H, m), 1.51-1.49 (1H, m), 1.42 (2H, brs), 1.37-1.35 (2H, m).

EXAMPLE 4

Preparation of N-[trans-4-[2-[4-(benzo[b]thiophene)-7-piperazinyl]ethyl]cyclohexyl]-N,N-dimethylurea (Compound 1)

(10) ##STR00027##

(11) 1.73 g of trans-4-[2-[4-(benzo[b]thiophene)-7-piperazinyl]ethyl]cyclohexylamine prepared in Example 3 was dissolved in 50 ml of dichloromethane. 1.40 ml of triethylamine was added, and then 5.50 mmol of N,N-dimethylcarbamyl chloride was added. The mixture was stirring for 48 hours at room temperature (25 C.2 C.). After the reaction was completed, 50 ml of water was added to extract and separate the reactant. The organic phase was concentrated (0.01 MPa, 45 C.) and the desired component was collected by using column chromatography of methanol:dichloromethane=1:10 (400 mesh silica gel) and then concentrated to give 1.89 g of the amorphous desired product.

(12) .sup.1H-NMR (CDCl.sub.3) ppm: 7.79 (1H, d, J=5.5 Hz), 7.76 (1H, d, J=8.1 Hz), 7.33 (1H, m), 7.28 (1H, d, 7.6 Hz), 6.96 (1H, d, J=7.6 Hz), 6.48 (1H, brs), 3.44-3.36 (8H, m), 3.58 (1H, m), 3.01 (6H, s), 2.46 (2H, m), 1.68-1.42 (8H, m), 1.52-1.48 (1H, m), 1.38-1.36 (2H, m).

EXAMPLE 5

Preparation of N-[trans-4-[2-[7-(benzo[b]thiophene)-7-piperazinyl]ethyl]cyclohexyl]-N,N-dimethylurea (Compound 2)

(13) Compound 2 was prepared according to the procedures given in Examples 1-4 by using 7-bromobenzo[b]thiophene as a starting material.

(14) .sup.1H-NMR (CDCl.sub.3) ppm: 7.78 (1H, d, J=5.6 Hz), 7.76 (1H, d, J=8.0 Hz), 7.31 (1H, m), 7.27 (1H, d, 7.6 Hz), 6.98 (1H, d, J=7.2 Hz), 6.44 (1H, brs), 3.48-3.42 (8H, m), 3.54 (1H, m), 3.00 (6H, s), 2.46 (2H, m), 1.68-1.42 (8H, m), 1.52-1.48 (1H, m), 1.38-1.36 (2H, m).

EXAMPLE 6

Preparation of N-[trans-4-[2-[4-(benzo[c]thiophene)-7-piperazinyl]ethyl]cyclohexyl]-N,N-dimethylurea (Compound 3)

(15) Compound 3 was prepared according to the procedures given in Examples 1-4 by using 4-bromobenzo[c]thiophene as a starting material.

(16) .sup.1H-NMR (CDCl.sub.3) ppm: 7.33 (2H, s), 7.27-7.25 (1H, m), 7.27 (1H, d, 7.6 Hz), 6.77 (1H, d, J=7.2 Hz), 6.73 (1H, d, J=7.2 Hz), 6.44 (1H, brs), 3.48-3.42 (8H, m), 3.54 (1H, m), 2.99 (6H, s), 2.46 (2H, m), 1.68-1.42 (8H, m), 1.52-1.48 (1H, m), 1.46-1.42 (2H, m).

EXAMPLE 7

Preparation of N-[trans-4-[2-[4-(6-fluoro-benzo[d]isoxazol)-3-piperazinyl]ethyl]cyclohexyl]-N,N-dimethylurea (compound 5)

(17) Compound 5 was prepared according to the procedures given in Examples 1-4 by using 6-fluoro-3-bromo-1,2 benzo[d]isoxazole as a starting material.

(18) .sup.1H-NMR (CDCl.sub.3) ppm: 7.41 (1H, d), 7.12 (1H, d), 6.98 (1H, s), 6.52 (1H, brs), 3.55 (1H, m), 3.46-3.42 (8H, m), 2.99 (6H, s), 2.45 (2H, m), 1.68-1.40 (8H, m), 1.50-1.48 (1H, m), 1.37-1.35 (2H, m).

EXAMPLE 8

Preparation of N-[trans-4-[2-[4-(3-chloro-benzo[d]isoxazol)-6-piperazinyl]ethyl]cyclohexyl]-N,N-dimethylurea (Compound 6)

(19) Compound 6 was prepared according to the procedures given in Examples 1-4 by using 3-chloro-6-bromo-benzo[d]isoxazole as a starting material.

(20) .sup.1H-NMR (CDCl.sub.3) ppm: 7.25 (1H, d), 6.78 (1H, s), 6.72 (1H, d), 6.51 (1H, brs), 3.54 (1H, m), 3.46-3.42 (8H, m), 2.99 (6H, s), 2.45 (2H, m), 1.67-1.40 (8H, m), 1.50-1.48 (1H, m), 1.42-1.35 (2H, m).

EXAMPLE 9

Preparation of N-[trans-4-[2-[4-(6-fluoro-benzo[d]isoxazol)-3-piperidyl]ethyl]cyclohexyl]-N,N-dimethylurea (Compound 7)

(21) ##STR00028##

(22) Compound 7 was prepared according to the procedures given in Examples 2-4 by using 6-fluoro-3-piperidin-4-yl-1,2 benzo[d]isoxazole (available from Shanghai Excellent Chemical Co., Ltd.) as a starting material.

(23) .sup.1H-NMR (CDCl.sub.3) ppm: 7.41 (1H, d), 7.12 (1H, d), 6.97 (1H, s), 6.51 (1H, brs), 3.55-3.53 (1H, m), 2.99 (6H, s), 2.78-2.76 (1H, m), 2.66-2.37 (4H, m), 2.46-2.40 (2H, m), 1.78-1.68 (12H, m), 1.50-1.48 (1H, m), 1.37-1.34 (2H, m).

EXAMPLE 10

Preparation of solid tablets of N-[trans-4-[2-[4-(benzo[b]thiophene)-7-piperazinyl]ethyl]cyclohexyl]-N,N-dimethylurea (Compound 1)

(24) 1000 tablets each weighing 200 mg were prepared according to the formulation shown in the table below.

(25) TABLE-US-00001 Amount Composition (g) Compound 1(active ingredient; 20.0 prepared in Example 4) Lactose 126.0 Microcrystalline Cellulose 42.0 Hydroxypropylmethyl Cellulose 4.0 Sodium Hydroxyethyl Starch 6.0 Magnesium Stearate 2.0

(26) The preparation method comprising: the active ingredient, lactose, microcrystalline cellulose and sodium hydroxyethyl starch were mixed and added to a high-shear wet granulator, stirring well at a certain rotating speed. Afterwards, 50.0 g of an aqueous solution of hydroxypropylmethylcellulose was added to the mixture to make it into appropriate granules under high-speed shearing conditions. The wet granules then were dried over a fluidized bed, and the resulting dried granules were uniformly mixed with the magnesium stearate and then compressed into tablets.

EXAMPLE 11

Preparation of an oral suspension of N-[trans-4-[2-[4-(benzo[b]thiophene)-7-piperazinyl]ethyl]cyclohexyl]-N,N-dimethylurea (Compound 1)

(27) 1000 bottles each containing 5 ml (10 mg/ml as per specification) were prepared according to the formulation shown in the table below.

(28) TABLE-US-00002 Amount Composition (g) Compound 1(active ingredient; 50.0 prepared in Example 4) water 3500.0 Polyethylene Glycol 50.0 Sorbitol 500.0 Microcrystalline Cellulose 25.0 Xanthan Gum 5.0 Methyl p-Hydroxybenzoate 1.25 Ethyl p-Hydroxybenzoate 1.25 Citrate Adjusting pH to 4-8

(29) The preparation method comprising: methyl p-hydroxybenzoate and ethyl p-hydroxybenzoate were dissolved in hot water and then cooled to room temperature (25 C.2 C.). Then, sorbitol, polyethylene glycol, xanthan gum, citrate, the compound 1 with an average particle size of 30 m and microcrystalline cellulose were successively added, and stirred well to give an oral suspension.

EXAMPLE 12

(30) Pharmacological Test

(31) I. In Vitro Test on Biological Activity of the Cyclohexane Derivative of the Present Invention.

(32) This test was carried out according to the dopamine D.sub.2/D.sub.3 receptor binding assay and the 5-HT.sub.1A receptor binding assay described in Jiangsu Hengyi Pharmaceutical Co., Ltd.'s Patent CN 103130737 A. IC50 values were calculated from concentration-dependent reactions using a non-linear analysis program. Ki values were calculated from IC50 values by using Cheng-Prussoff equation, i.e. Ki=IC.sub.50/(1+[L]/KD), wherein Ki is the affinity of the drug to the receptor; L is the concentration of the compound to be tested; and KD is the affinity of radioligand to the receptor.

(33) (I) Dopamine D.sub.2 Receptor Binding Assay

(34) i. Materials

(35) a. Transfection of D.sub.2 Receptor Cells: by using calcium phosphate method, HEK293 cells were transfected with a plasmid vector containing the D.sub.2 receptor protein gene. The transfected cells were cultured in a culture solution containing G418, followed by a selection of monoclonal cell and radioligand binding assay, finally obtaining a stable cell line with stable expression of D.sub.2 receptor protein. b. Materials for receptor binding assay: isotope ligand [.sup.3H] Spiperone (113.0 Ci/mmol); available from Sigma-Aldrich Company; (+) spiperone, available from RBI company; GF/B glass fiber filter paper, available from Whatman Company; Tris imported and packaged; PPO, POPOP, available from Shanghai Reagent Chemical Company; lipid-soluble scintillation solution; and Beckman LS-6500 Multi-function Liquid Scintillation Counter.
ii. Test Method a. Treatment of cells: HEK-293 cells were infected with various recombinant viruses of the above genes, respectively, and the receptor proteins were highly expressed on the membranes after 48-72 hours. The cells were centrifuged at 1000 rpm for 5 minutes. The culture solution was discarded, and the cells were collected and stored in a refrigerator at 20 C. for standby. The cells were resuspended with Tris-HCl buffer (pH=7.5) when tested. b. Competitive binding assay for receptor

(36) The compound to be tested and the radio ligand, both 20 L, and 160 L of receptor proteins were added to the reaction test tubes to achieve a final concentration of 10 mol/L for each of the compound to be tested and positive control, incubated in a water bath of 30 C. for 50 minutes and immediately transferred to an ice bath to terminate the reaction; placed on a Millipore cell sample collector, filtered by suction rapidly through a GF/C glass fiber filter, and eluted with 3 mL of eluent (50 mM Tris-HCl, pH7.5) for three times and dried with microwave for 5-6 minutes. The filter paper was transferred into a 0.5 mL centrifuge tube to which 500 L of lipid-soluble scintillation solution was added, and settled away from light for more than 30 minutes. The radioactive intensity was determined by counting. The concentration of the compound was 10 mol/L, and inhibition rate (%) of each compound to the binding of isotope ligands was calculated according to the following equation:
Inhibition rate (I%)=(total binding tube cpmcompound cpm)/(total binding tube cpmnon-specific binding tube cpm)100%.
(II) Dopamine D.sub.3 Receptor Binding Assay

(37) The concentration of the compound was 10 mol/L, and the assay was performed according to the method described in Journal of Pharmacology and Experimental Therapeutics 2010, 333 (1): 328.

(38) (III) 5-HT.sub.1A Receptor Binding Assay

(39) i. Materials

(40) 5-HT.sub.1A receptor isotope ligand [.sup.3H] 8-OH-DPAT (available from PE Company); (+)5-hydroxytrptamine (available from Sigma Company); GF/B glass fiber filter paper (available from Whatman Company); lipid-soluble scintillation solution: PPO, POPOP (available from Shanghai Chemical Reagent Company); toluene (from Sinopharm Chemical Reagent Co., Ltd.); Tris imported and packaged.

(41) Treatment of cells: HEK-293 cells which stably express the 5-HT.sub.1A receptor by gene recombination were cultured in DMEM+10% serum solution for 3-5 days and then collected with PBS. The cells were centrifuged at 3000 rpm and 4 C. for 10 minutes. Afterwards, the supernate was discarded, and the cells were collected and stored in a refrigerator at 80 C. The cells were resuspended with D.sub.1 Binding Buffer (pH 7.4) when tested.

(42) ii. Test Method

(43) Inhibition rate of each compound of 10 umol/L to the binding of [.sup.3H]8-OH-DPAT and the 5-HT.sub.1A receptor was determined by a general selection.

(44) The experimental data is shown in Table 1.

(45) TABLE-US-00003 TABLE 1 Binding Assay of Compounds to D.sub.2 and D.sub.3 Receptors and Affinity to 5-HT.sub.1A Receptor (Ki: nmol) Compounds D.sub.2 D.sub.3 5-HT.sub.1A >1000 0.06 3.85 0 978.2 0.07 3.72 >1000 0.06 3.96 1.3 0.29 4.12 250.9 0.57 22.83 >1000 0.15 3.65

(46) Conclusion: it can be seen from the experimental results in Table 1 that the series of compounds of the present invention have a strong affinity to D.sub.3 and a very weak affinity to D.sub.2 with nearly ten thousand folds difference between them, demonstrating that the series of compounds have high selectivity to D.sub.2/D.sub.3 receptors and reduce side effects when selecting D.sub.2 receptor. Further, the compounds show relatively strong affinity to 5-HT.sub.1A receptor and act on a wide spectrum of neuropsychiatric diseases.

EXAMPLE 13

(47) In Vivo Anti-Schizophrenic Activity Assay of the Cyclohexane Derivatives of the Present Invention

(48) I. MK-801 Model

(49) (I) Modeling of MK-801-Induced Schizophrenia in Mice

(50) 100 Sprague-Dawley rats (provided by Shanghai Lake Experimental Animal Co., Ltd.), all males, were randomly divided into 10 groups according to body weights: blank control, MK-801 model control, the cyclohexane derivatives C.sub.1 to C.sub.8 (corresponding to the compound 1 to the compound 7 and compound 14; 0.3 mg/kg) and Cariprazine positive control (prepared according to the method described in Patent CN103130737A; 0.3 mg/kg). Each rat was placed in a soundproof box for 30 minutes on the day before the test to adapt. On the next day, each rat was administrated with the respective test compounds, and after 30 minutes, administrated intraperitoneally with a 0.3 mg/kg MK-801 solution at 5.0 mL/kg body weight of the rat to build a model of MK-801-induced schizophrenia in mice.

(51) Administration: the rats in the present cyclohexane derivatives and the cariprazine positive control groups were orally administered (i.g.), while the MK-801 model control group was administered intraperitoneally.

(52) (II) Observation of Open Field Running Behavior

(53) Mice were administered with MK-801 and then immediately placed in the soundproof box. The total distances of motion of mice within 2.5 hours were observed and recorded.
Improvement rate=(total distance of model controltotal distance of administration)/(total distance of model control)100%.
(III) Statistical Method

(54) All the data was expressed with xSD and processed by SPSS17.0 software statistical package. T-test and one-way analysis of variance were performed to compare the mean of two samples, with P<0.05 as significant difference.

(55) (IV) Results

(56) The results specifically are shown in Table 2 below.

(57) TABLE-US-00004 TABLE 2 Effect of Single Oral Administration of the Cyclohexane Derivatives on the Total Distance of Open Field Motion in MK-801-Induced Schizophrenia in Mice Model (x SD) Total Distance Improve- Num- within 61-150 ment ber of Dosage minutes Rate Groups Rats (mg/kg) (m) (%) Blank Control 10 MK-801Model 10 0.3 mg/kg 309.78 39.1 Control Cariprazine 10 0.3 mg/kg 168.25 26.9**** 45.69 Positive Control C1 (Compound 1) 10 0.3 mg/kg 73.09 11.7**** 76.41 C2 (Compound 2) 10 0.3 mg/kg 46.95 10.3**** 84.84 C3 (Compound 3) 10 0.3 mg/kg 63.79 10.8**** 79.41 C4 (Compound 4) 10 0.3 mg/kg 232.18 20.3* 25.05 CS (Compound 5) 10 0.3 mg/kg 5.43 1.7**** 98.25 C6 (Compound 6) 10 0.3 mg/kg 171.78 13.0* 12.27 C7 (Compound 7) 10 0.3 mg/kg 160.82 12.91* 48.09 C8 (Compound 14) 10 0.3 mg/kg 150.28 11.70* 51.49

(58) The results in Table 2 show that the cariprazine positive control and the cyclohexane derivatives (Compound 1 to Compound 7) of the present invention have decreased the total distances of the rats within 150 minutes as compared with the MK-801 model control. Wherein *P<0.05, ****P<0.0001.

(59) These results demonstrate that: i. compared with the blank control, the distance of the open-field motion of MK-801 model control has increased significantly, indicating that MK-801 may causes schizophrenia in mice; ii. compared with the MK-801 model control, and the cariprazine positive control has significantly inhibited the high spontaneous activity of MK-801-induced rats (P <0.0001) at a dose of 0.3 mg/kg; the cyclohexane derivatives, Compounds 1, 2, 3 and 5 of the present invention can significantly inhibit the high spontaneous activity of MK-801-induced rats (P<0.0001) at a dose of 0.3 mg/kg, and the cyclohexane derivatives, Compounds 6, 7 and 14 of the present invention also have significantly inhibited the high spontaneous activity of MK-801-induced rats (P<0.05) at a dose of 0.3 mg/kg , which are comparable with the cariprazine positive control. Since the MK-801-induced open-field motion model is closely related to the symptoms of schizophrenia, it is indicated that the cyclohexane derivative series of the present invention have a significant effect on schizophrenia.

EXAMPLE 14

(60) Acute Toxicity Assay of the Cyclohexane Derivatives of the Present Invention

(61) ICR mice orally administered with the compounds of the present invention were evaluated and their toxicity symptoms after oral administration were observed, as well as the mortality, which was calculated by Bliss method, thus the acute toxicities were compared.

(62) Experimental Program

(63) i. Preparation of solvent: Tween-80 of an appropriate amount was diluted with deionized water to a concentration of 5% (g/v) Tween-80. ii. Formulations for administration: test samples were weighed as required, respectively, and made into suspensions with 5% the Tween-80 solution to achieve concentrations of 0.94, 1.88, 3.75, 7.5, 15, 30 and 60 mg/mL, respectively (equivalent to 18.75, 37.5, 150, 300, 600 and 1200 mg/kg, respectively). iii. Administration: the test compound and solvent medium control groups (0.5% Tween-80) were administered orally.

(64) Observation of general symptoms: Day 1: rats were observed at about 10 minutes, 0.5, 2, 4 and 6 hours after the first administration, respectively; Days 2-6, mice were observed twice a day, once in the morning and once in the afternoon. Observations include but are not limited to: general condition, behavior, gait, eyes, mouth, nose, gastrointestinal tract, skin and hair and urogenital tract.

(65) The experimental results are shown in Table 4.

(66) TABLE-US-00005 TABLE 4 Acute Toxicity of Single Oral Administration of Compounds Compounds LD.sub.50 (mg/Kg) 1100 mg/KG 1050 mg/KG 1080 mg/KG

(67) Conclusion: the acute toxicity of the cyclohexane derivatives of the present invention is far lower than that of the cariprazine positive control (RGH-188, 75.3 mg/kg), showing good safety.