WO2017045599A1 - Cyclohexane derivative or stereoisomer or salt thereof, and preparation and use thereof - Google Patents

Abstract

Provided are a cyclohexane derivative as shown by formula IB or a stereoisomer or a salt thereof, and the preparation and use thereof. The cyclohexane derivative has a high affinity for D₃ receptors and 5-hydroxytryptamine, has a lower affinity for D₂ receptors, shows a high selectivity for D₃/D₂ receptors, and can be used as a therapeutic drug against neuropsychiatric diseases; and the preparation method thereof is simple and easy.

Description

Cyclohexane derivative or stereoisomer or salt thereof and preparation and application thereof Technical field

The present invention relates to medicinal chemistry, in particular to cyclohexane compounds or stereoisomers or salts thereof, and in particular to cyclohexane derivatives of formula IB and formula I or stereoisomers or salts thereof, and their preparation and application.

Background technique

Along with the rapid development of society, people's life rhythm and pressure are increasing. Mental diseases have become a serious disease affecting human health, which has serious consequences for patients and their families. The average life expectancy of patients is reduced due to suicide, lack of medical care, and high risk of complications such as malnutrition, lack of exercise, obesity and smoking. Numerous studies have shown that mental illness is associated with a variety of neurotransmitters and receptor dysfunction in the central, such as monoamine transmitters in the brain, especially dopamine (DA) systems and serotonin (5-HT) systems and normal mental activity in humans. closely related. When the DA and 5-HT systems are dysfunctional, they are likely to cause a variety of neuropsychiatric diseases such as schizophrenia, depression, neuropathic pain, mania, anxiety, and Parkinson's disease.

Patent WO 9967206 A1 describes the use of cyclohexane derivatives in the treatment of pain disorders, but does not publicly report their use in psychotic diseases, especially on the dopamine D ₂ /D ₃ receptor.

Patent CN 1829703A describes the use of a cyclohexane derivative having a (thio)carbamoyl side chain in the regulation of a dopamine receptor disorder, wherein the D ₂ /D ₃ antagonist and 5 jointly developed by Forest Laboratories and Gedeon Richter - The HT _1A partial agonist, Cariprazine (RGH-188), has been completed in clinical trials for the treatment of schizophrenia and mania and depression. The chemical structure of the cariprazine is shown in the following formula. The affinity (Ki value) of the drug for the D ₂ /D ₃ receptor and 5-HT _1A is 0.72 nmol, 0.08 nmol and 3.42 nmol, respectively, for D ₂ /D. Although the ₃ receptor has certain selectivity, it is still not ideal, so it may cause the drug to have a low probability of clinically (approximately 5% probability at 3 mg dose) of sedation and the cause of extracorporeal reaction. Because these side effects are associated with excessive blockade of the D ₂ receptor.

Based on the above problems, the patent CN 103130737A has further structural modification of the cariprazine to achieve higher D ₃ receptor selectivity.

However, there are many principles for the generation of mental diseases. Although the above compounds have good efficacy against schizophrenia, they cannot meet the needs of the treatment of mental diseases, and should be further researched and developed.

Summary of the invention

The technical problem to be solved by the present invention is to overcome the above-mentioned deficiencies, and to study and design to improve the structure of a cyclohexane derivative, and to provide a cyclohexane derivative or a stereoisomer thereof or a salt thereof as shown in Formula IB and Formula I, It has D ₂ /D ₃ antagonist action and serotonin absorption inhibition, as well as anti-schizophrenia, increases the broad spectrum of treatment for psychotic diseases, and reduces side effects.

The cyclohexane derivative or a stereoisomer or salt thereof provided by the present invention has the structure shown in the following formula IB:

Where X is N or C;

R is

And, the R group may be optionally substituted by one or more substituents selected from halogen, substituted or unsubstituted C ₁ -C ₆ alkyl; the halogen is selected from F, Cl, One or more of Br or I; the substituted or unsubstituted C ₁ -C ₆ alkyl group is selected from a substituted or unsubstituted C ₁ -C ₄ alkyl group, such as methyl, ethyl, propyl or Butyl, the substituent is a halogen, such as one or more of F, Cl, Br or I, and the substituted C ₁ -C ₄ alkyl group is preferably a trifluoromethyl group.

Preferably, the cyclohexane derivative provided by the present invention, or a stereoisomer or salt thereof, has the structure shown in the following formula I:

Where R is

And, the R group may be optionally substituted by one or more substituents selected from halogen, substituted or unsubstituted C ₁ -C ₆ alkyl; the halogen is selected from F, Cl, One or more of Br or I; the substituted or unsubstituted C ₁ -C ₆ alkyl group is selected from a substituted or unsubstituted C ₁ -C ₄ alkyl group, such as methyl, ethyl, propyl or Butyl, the substituent is a halogen, such as one or more of F, Cl, Br or I, and the substituted C ₁ -C ₄ alkyl group is preferably a trifluoromethyl group.

The stereoisomer of the cyclohexane derivative of the present invention is a cis stereoisomer or a trans stereoisomer, preferably a trans stereoisomer.

The salt of the cyclohexane derivative of the present invention is formed from a cyclohexane derivative which is an organic acid or an inorganic acid, and the inorganic acid is selected from hydrochloric acid, sulfuric acid, nitric acid or phosphoric acid; From formic acid, acetic acid, oxalic acid malonic acid, maleic acid, fumaric acid, succinic acid or benzoic acid; and other physiologically acceptable salts.

Preferably, the cyclohexane derivative of the present invention or a stereoisomer or salt thereof is selected from the following compounds or salts thereof:

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-(benzo[d]isothiazole)-3-piperazinyl]ethyl]cyclohexyl]-N,N-dimethylurea (compound 4)

N'-[trans-4-[2-[4-(6-fluoro-benzo[d]isoxazole)-3-piperazinyl]ethyl]cyclohexyl]-N,N-dimethyl Urea (compound 5)

N'-[trans-4-[2-[4-(3-chloro-benzo[d]isoxazole)-6-piperazinyl]ethyl]cyclohexyl]-N,N-dimethyl Urea (compound 6)

N'-[trans-4-[2-[4-(6-fluoro-benzo[d]isoxazole)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]isothiazole)-3-piperidinyl]ethyl]cyclohexyl]-N,N-dimethylurea (compound 11)

N'-[trans-4-[2-[4-(3-chloro-benzo[d]isoxazole)-6-piperidinyl]ethyl]cyclohexyl]-N,N-dimethyl Urea (compound 12)

N'-[trans-4-[2-[4-(3-methyl-benzo[d]isoxazole)-6-piperazinyl]ethyl]cyclohexyl]-N,N-dimethyl Base urea (compound 13)

N'-[trans-4-[2-[4-(6-methyl-benzo[d]isoxazole)-4-piperazinyl]ethyl]cyclohexyl]-N,N-dimethyl Base urea (compound 14)

Another object of the present invention is to provide a process for the preparation of the cyclohexane derivative, or a stereoisomer or salt thereof, which comprises the steps of:

4-ethylcyclohexylamine derivative II is reacted with N,N-diformylcarbonyl chloride III in the presence of an acid binding agent to give IB compound IB:

Where R and X are the same as defined above.

The molar ratio of the compound II to the compound III is 1-1.5:1, the reaction temperature is 0 ° C - 50 ° C, the acid binding agent is an organic basic substance or an inorganic basic substance; the organic basic substance is selected from the group consisting of three One or more of an amine, diisopropylethylamine or pyridine selected from one or more of sodium carbonate, potassium carbonate, sodium hydrogencarbonate or potassium hydrogencarbonate, the binding The molar ratio of acid agent (basic compound) to compound II is 1-1.5:1.

The invention also provides a preparation method of the compound II, the method comprising the following steps:

or

Wherein R and X are as defined above, and Pg is an amino protecting group selected from the group consisting of benzyl Bn, benzyl formate CBz or t-butoxycarbonyl Boc. The compound of formula II may itself be cis or trans, or the compound of formula IB may be finally chromatographed or crystallized to obtain its cis stereoisomer.

(1) When X is nitrogen, a piperazine and a bromine are subjected to a coupling reaction to obtain an intermediate IV;

The coupling reaction is carried out under the palladium catalysis, in the presence of a strong base potassium t-butoxide, sodium t-butoxide, potassium carbonate or cesium carbonate, the reaction temperature is 50-150 ° C, the molar ratio of piperazine to bromine 1-5:1; or,

When X is C, the intermediate IV is commercially available (Shanghai Key Chemicals).

(2) Intermediate IV and 4-amino protected cyclohexane acetaldehyde are subjected to condensation imine reduction reaction to obtain intermediate V;

The reducing agent is a boron compound such as sodium triacetoxyborohydride or sodium borohydride; the molar ratio of the intermediate IV to the 4-amino protected cyclohexaneacetaldehyde is 1-1.2:1; the boron compound and the intermediate The body IV molar ratio is 1-2:1;

(3) a deamination protecting group;

More specifically, when Pg is t-butoxycarbonyl Boc, an acid deamination protecting group is added; the acid is an organic solution of hydrogen chloride or trifluoroacetic acid; or, when Pg is benzyl Bn or benzyl formate CBz, A palladium carbon hydrodeamination protecting group is used, and the hydrogenation pressure is from 0.1 to 1 MPa.

A further object of the present invention is to provide use of the cyclohexane derivative or a stereoisomer or salt thereof for the preparation of a medicament for anti-neuropsychiatric diseases.

The cyclohexane derivative or the stereoisomer or salt thereof according to the present invention is a drug-effect structure having potential action on dopamine D ₃ receptor and has potential serotonin absorption according to the design principle of the compound pharmacodynamic structure fusion drug. Inhibition of the pharmacodynamic structure of the fusion, through the structural modification of the compound, preparation, in vitro activity test, in vivo anti-schizophrenia activity test, structure-activity relationship study and re-optimization of new compounds. The results of pharmacological studies indicate that the cyclohexane derivative of the present invention or a stereoisomer or salt thereof has a strong affinity for the D ₃ receptor and serotonin, and has a weak affinity for the D ₂ receptor, showing a pair D. The high selectivity of the ₃ /D ₂ receptor achieves unexpected results. The in vitro receptor binding assay showed that most of the compounds involved in the cyclohexane derivative or its stereoisomer or salt of the present invention have a strong affinity for the dopamine D ₃ receptor and the 5-HT _1A receptor (Ki<10 nmol). ), a weak affinity for the dopamine D ₂ receptor (Ki > 50 nmol), exhibiting good D ₃ /D ₂ receptor selectivity, and strong affinity for the 5-HT _1A receptor. The in vivo anti-schizophrenia activity test showed that the cyclohexane derivative of the present invention or a stereoisomer or salt thereof has a strong anti-schizophrenia symptom. Structure-activity relationship studies show strong affinity for dopamine D ₃ receptor, 5-HT _1A receptor and high selectivity to D ₃ /D ₂ receptor, as well as anti-schizophrenia symptoms, and the present invention The benzoheterocyclic fragment such as benzothiophene, benzisothiazole or benzisoxazole fragment in the structure of the cyclohexane derivative series compound is closely related (the corresponding fragment of the existing carilazide is 2, 3 chlorobenzene).

Therefore, the cyclohexane derivative of the present invention or a stereoisomer or salt thereof can be used for the preparation of a neuropsychiatric drug.

The medicament according to the present invention means a pharmaceutical composition comprising the cyclohexane derivative or a stereoisomer or salt thereof as an active ingredient and a pharmaceutically acceptable adjuvant.

The pharmaceutical composition of the present invention can be selected for any convenient mode of administration. For example, oral, gastrointestinal, buccal, sublingual, nasal, rectal or transdermal administration. It can be developed into different dosage forms such as solid dosage forms and liquid dosage forms such as suspensions, tablets, capsules.

The composition in the form of a solid tablet may contain a filler, a lubricant, a binder, a disintegrator conventionally used in the formulation. The liquid preparation is a liquid carrier such as a water-soluble solvent such as water, ethanol or glycerin, or a water-insoluble solvent such as polyethylene glycol or a suspension or solution in an oil.

More specifically, the pharmaceutical composition of the present invention is a solid tablet comprising a cyclohexane derivative of the formula IB or the formula I or a stereoisomer or salt thereof as an active ingredient and a pharmaceutically acceptable adjuvant.

The solid tablet consists of the following weight ratio components:

The diluent is selected from the group consisting of starch, lactose or microcrystalline cellulose; the binder is selected from the group consisting of hydroxybenzylcellulose, hydroxypropylmethylcellulose or polyvinylpyrrolidone; the disintegrant is selected from hydroxyB Based on sodium starch or crospovidone, the lubricant is magnesium stearate.

Preferably, the present invention provides N'-[trans-4-[2-[4-(benzo[b]thiophene)-7-piperazinyl]ethyl]cyclohexyl]-N,N-dimethyl A base urea compound is used as a solid tablet composed of an active ingredient and a pharmaceutically acceptable adjuvant.

The solid tablet consists of the following weight ratio components:

Wherein the diluent is selected from the group consisting of starch, lactose or microcrystalline cellulose, the binder is selected from the group consisting of hydroxybenzylcellulose, hydroxypropylmethylcellulose or polyvinylpyrrolidone, and the disintegrant is selected from sodium hydroxyethyl starch or Dividone, the lubricant is magnesium stearate.

Alternatively, the pharmaceutical composition of the present invention is a suspension comprising a cyclohexane derivative of the formula IB or the formula I or a stereoisomer or salt thereof as an active ingredient and a pharmaceutically acceptable adjuvant.

The suspension consists of the following weight ratio components:

The suspending agent is selected from the group consisting of xanthan gum or microcrystalline cellulose; the preservative is selected from sodium benzoate, methyl p-hydroxybenzoate or ethyl p-hydroxybenzoate, the diluent being selected from water or sorbitol; The buffering agent is a citrate; the cosolvent is selected from the group consisting of cyclodextrin, ethanol, propylene glycol or polyethylene glycol; the flavoring agent may be a sweetener (such as sugar, saccharin, etc.) well known to those skilled in the art; The agent may be a fat-soluble colorant or a water-soluble colorant such as carotene, Cocoa pigment, caramel color, etc.

Preferably, the present invention provides N'-[trans-4-[2-[4-(benzo[b]thiophene)-7-piperazinyl]ethyl]cyclohexyl]-N,N-dimethyl A base urea compound is used as a suspension of the active ingredient and a pharmaceutical excipient.

The suspension consists of the following weight ratio components:

The suspending agent is selected from the group consisting of xanthan gum or microcrystalline cellulose, and the preservative is selected from the group consisting of sodium benzoate, methyl p-hydroxybenzoate, ethyl p-hydroxybenzoate, the diluent is selected from the group consisting of water, sorbitol, and the buffer is lemon. The acid salt, the co-solvent is selected from the group consisting of cyclodextrin, ethanol, propylene glycol or polyethylene glycol, the flavoring agent may be a sweetener (such as sugar, saccharin, etc.) well known to those skilled in the art; the coloring agent is a fat-soluble coloring agent or water-soluble. Sexual coloring agents such as carotene, cocoa pigment or caramel coloring.

It is still another object of the present invention to provide the use of the cyclohexane derivative or a stereoisomer or salt thereof for the preparation of a medicament for anti-neuropsychiatric diseases, which application refers to preparation for improvement/treatment of neuropsychiatric diseases A drug that can be used to prepare drugs that improve/treat schizophrenia, mental disorders, mental disorders, mental disorders, mood disorders, bipolar disorder, depression, phobia, obsessive-compulsive disorder, anxiety disorder or cognitive disorder .

Acute toxicity experiments show that the cyclohexane derivative or its stereoisomer or salt of the present invention has only very low toxicity (LD ₅₀ >1000 mg/Kg), which is superior to Cariprazine (LD). ₅₀ = 75.3 mg / Kg). The drug of the invention has low toxicity and good safety.

Pharmacological tests have shown that the cyclohexane derivative of the present invention or a stereoisomer or salt thereof is a novel therapeutic drug for anti-neuropsychiatric diseases, has a good clinical application prospect, and brings good news to patients. Social benefits. Moreover, the preparation method of the compound of the invention is simple and easy, and is suitable for industrial production, and has great application value.

detailed description

The materials and reagents used in the following examples are commercially available unless otherwise specified.

Example 1

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

7.20 g of 4-bromobenzo[b]thiophene, 19.9 g of piperazine anhydride, 4.70 g of sodium t-butoxide, 0.32 g of (R)-(+)-2,2'-bis(diphenylphosphino)- A mixture of 1,1'-binaphthalene (BINAP), 0.63 g of dipalladium tris(dibenzylideneacetone) and 150 ml of toluene was refluxed for 1 hour under a nitrogen atmosphere. 150 ml of water was poured into the reaction solution, and the mixture was extracted with 100 ml of ethyl acetate. After washing with water, dried over anhydrous magnesium sulfate, and the solvent was evaporated under reduced pressure (0.01 MPa, 45 ° C). The residue was purified by silica gel column chromatography (dichlorobenzene:methanol:EtOAc:EtOAc: EtOAc) 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 the solvent was evaporated under reduced pressure (0.01 MPa, 45 ° C). Ethyl acetate (50 ml) was added to the residue, and the crystals were crystallized, filtered, dissolved in 15 ml of methanol, and then cooled to room temperature (25 ° C) to recrystallize to give 1-benzene[b]thiophene as colorless needle crystals. 4-yl-piperazine hydrochloride.

Example 2

Preparation of trans-4-[2-[4-(benzo[b]thiophene)-7-piperazinyl]ethyl]cyclohexyl-carbamic acid tert-butyl ester

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-un Butyroxycarbyl)amino]cyclohexyl}-acetaldehyde was dissolved in 120 ml of dichloromethane, and 1.40 ml (10 mmol) of triethylamine was added at room temperature (25 ° C ± 2 ° C) for 10 minutes, then gradually added 3.16 g. (14.8 mmol) sodium triacetoxyborohydride, the reaction was further stirred at room temperature for 24 hours, and after completion of the reaction, 120 ml of a 10% sodium hydrogencarbonate solution was added. The reaction system was directly subjected to extraction and liquid separation. The organic phase was dried over anhydrous sodium sulfate, and then filtered and evaporated to dryness. The solid was refluxed with 15 ml of ethyl acetate and cooled to room temperature (25 ° C ± 2 ° C) to give 3.70 g of the desired product.

¹ H-NMR (CDCl ₃ ) δ 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

4.43 g of trans-4-[2-[4-(benzo[b]thiophene)-7-piperazinyl]ethyl]cyclohexyl-carbamic acid tert-butyl ester (obtained in Example 2) under ice-water bath The reaction mixture was placed in a reaction flask, and 80 ml of a saturated aqueous solution of hydrogen chloride in ethyl acetate was added, and the reaction was stirred for 8 hours to carry out a deprotection reaction to give a white solid. The solid was added to 50 ml of a dichloromethane solution, and 50 ml of a saturated sodium hydrogencarbonate solution was stirred for half an hour, and then the mixture was separated and extracted, and the organic phase was concentrated (0.01 MPa, 40 ° C) to give the object product 3.30 g.

¹ H-NMR (CDCl ₃ ) δ 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

N'-[trans-4-[2-[4-(benzo[b]thiophene)-7-piperazinyl]ethyl]cyclohexyl]-N,N-dimethylurea (compound 1) preparation

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 followed by 5.50 mmol of N,N-diformylcarbonyl chloride. Stir at room temperature (25 ° C ± 2 ° C) for 48 hours. At the end of the reaction, 50 ml of water was added to extract the liquid, and the organic phase was concentrated (0.01 MPa, The target component was collected by a methanol:dichloromethane = 1:10 column chromatography (400 mesh silica gel model), and concentrated to give 1.89 g of the objective product.

¹ H-NMR (CDCl ₃ ) δ 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

N'-[trans-4-[2-[7-(benzo[b]thiophene)-7-piperazinyl]ethyl]cyclohexyl]-N,N-dimethylurea (compound 2) preparation

7-Bromobenzo[b]thiophene was prepared as a starting material, and Compound 2 was prepared by the same procedure as in Example 1-4.

¹ H-NMR (CDCl ₃ ) δ 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

N'-[trans-4-[2-[4-(benzo[C]thiophene)-7-piperazinyl]ethyl]cyclohexyl]-N,N-dimethylurea (compound 3) preparation

Starting from 4-bromobenzo[c]thiophene, compound 3 was prepared by following the procedures of Examples 1-4.

¹ H-NMR (CDCl ₃ ) δ 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

N'-[trans-4-[2-[4-(6-fluoro-benzo[d]isoxazole)-4-piperazinyl]ethyl]cyclohexyl]-N,N-dimethyl Preparation of urea (compound 5)

Starting from 6-fluoro-3-bromo-1,2benzo[d]isoxazole, compound 5 was prepared by the same procedure as in Example 1-4.

¹ H-NMR (CDCl ₃ ) δ 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

N'-[trans-4-[2-[4-(3-chloro-benzo[d]isoxazole)-6-piperazinyl]ethyl]cyclohexyl]-N,N-dimethyl Preparation of urea (compound 6)

Prepared by 3-chloro-6-bromo-benzo[d]isoxazole as starting material, compound 6 was prepared according to the procedure of Example 1-4.

¹ H-NMR (CDCl ₃ ) δ 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

N'-[trans-4-[2-[4-(6-fluoro-benzo[d]isoxazole)-4-piperidinyl]ethyl]cyclohexyl]-N,N-dimethyl Preparation of urea (compound 7)

Starting from 6-fluoro-3-piperidin-4-yl-1,2benzo[d]isoxazole (purchased from Shanghai Key Chemicals), compound 7 was prepared by the same procedure as in Example 2-4.

¹ H-NMR (CDCl ₃ ) δ 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

Solid tablet of N'-[trans-4-[2-[4-(benzo[b]thiophene)-7-piperazinyl]ethyl]cyclohexyl]-N,N-dimethylurea compound 1 Agent

1000 tablets were prepared according to the formulation in the table below, each weighing 200 mg.

Composition	Dosage (g)
Compound 1 (active ingredient) obtained in Example 4	20.0
lactose	126.0
Microcrystalline cellulose	42.0
Hydroxypropylmethylcellulose	4.0
Sodium hydroxyethyl starch	6.0
Magnesium stearate	2.0

Preparation method: the active ingredient, lactose, microcrystalline cellulose and sodium hydroxyethyl starch are mixed, added to a high shear wet granulator, and stirred uniformly at a certain rotation speed; then an aqueous solution of hydroxypropylmethylcellulose is added to the mixture. 50.0 g, suitable pellets were prepared under high shear conditions; then the wet granules were dried by a fluidized bed; the obtained dry granules were uniformly mixed with magnesium stearate and tableted.

Example 11

Oral mixing of N'-[trans-4-[2-[4-(benzo[b]thiophene)-7-piperazinyl]ethyl]cyclohexyl]-N,N-dimethylurea compound 1 Suspension

1000 bottles of 5 ml each were prepared according to the formulation of the oral suspension in the table below (according to the specification 10 mg/ml).

Composition	Dosage (g)
Compound 1 (active ingredient) obtained in Example 4	50.0
water	3500.0
Polyethylene glycol	50.0
Sorbitol	500.0
Microcrystalline cellulose	25.0
Xanthan gum	5.0
Methylparaben	1.25
Ethyl p-hydroxybenzoate	1.25
Citrate	Adjust pH 4~8

Preparation method: methyl p-hydroxybenzoate and ethyl p-hydroxybenzoate are dissolved in hot water, cooled to room temperature (25 ° C ± 2 ° C), and then added sorbitol, polyethylene glycol, xanthan gum, citric acid The salt, the compound 1 having an average particle size of 30 μm, and the microcrystalline cellulose were uniformly stirred to obtain an oral suspension.

Example 12 Pharmacological Experiment

I. In vitro biological activity of the cyclohexane derivative of the present invention

Pharmacological experiments were carried out according to the dopamine D ₂ /D ₃ receptor binding assay described in Jiangsu Hengyi Pharmaceutical Co., Ltd. patent CN 103130737A and the 5-HT _1A receptor binding assay. Non-linear analysis program from the concentration - dependent response ₅₀ calculated value IC. The Ki value was calculated from the IC ₅₀ value using the Cheng-Prussoff formula. That is: Ki = IC ₅₀ /(1+[L]/KD), Ki is the affinity of the drug to the receptor, L is the concentration of the test compound, and KD is the affinity of the radioligand to the receptor.

(a) Dopamine D ₂ receptor binding assay

1. Experimental materials:

1. D ₂ receptor cell transfection: HEK293 cells were transfected with a plasmid vector containing the D ₂ receptor protein gene, and subjected to calcium phosphate transfection, and cultured from the transfected cells through a medium containing G418. As well as the selection of cell monoclonal and radioligand binding experiments, a stable cell line stably expressing the D ₂ receptor protein was finally obtained.

2. Receptor binding experimental materials: isotopic ligand [ ³ H]Spiperone (113.0 Ci/mmol); purchased from Sigma; (+) spiperone, purchased from RBI; GF/B glass fiber filter paper, purchased from Whatman; Tris imported sub-package; PPO, POPOP purchased from Shanghai Reagent No. 1; fat-soluble scintillation liquid. Beckman LS-6500 multi-function liquid scintillation counter.

2. Experimental methods:

1. Cells: HEK-293 cells were infected with recombinant viruses containing various genes. After 48-72 hours, the receptor protein was expressed in a large amount on the membrane. After centrifugation at 1000 rpm for 5 min, the cells were discarded, and the cells were collected and stored in - Spare in the refrigerator at 20 °C. The experiment was resuspended in Tris-HCl reaction buffer (pH = 7.5).

2. Receptor competition binding experiment:

Add 20 μL of the test compound and the radioactive ligand to the reaction tube, and add the final concentration of the test compound and the positive drug to 10 μmol/L. After incubating in a 30 ° C water bath for 50 min, immediately transfer to an ice bath to terminate the reaction. On a Millipore cell sample collector, rapidly filtered through GF/C glass fiber filter paper, and eluate (50 mM Tris-HCl, pH 7.5) 3 mL × 3 times, dried in a microwave for 5-6 min, and the filter paper was transferred to 0.5 mL. Add 500 μL of fat-soluble scintillation fluid to the centrifuge tube. The light was allowed to stand for 30 minutes or more, and the radioactivity was measured by counting. The compound concentration was 10 umol/L, and the percentage inhibition rate of each compound for isotope ligand binding was calculated according to the following formula:

Inhibition rate (I%) = total binding tube cpm - compound cpm / total binding tube cpm - non-specific binding tube cpm x 100%.

(B) dopamine D ₃ receptor binding test

Compound 10 umol/L concentration, the experimental method was carried out in accordance with Journal of Pharmacology and Experimental Therapeutics 2010, 333(1):328.

(C) 5-HT _1A receptor binding assay

1. Experimental materials:

5-HT _1A receptor isotope [ ³ H].8-OH-DPAT (purchased from PE), (+) 5-hydroxytrptamine (purchased from Sigma), GF/B glass fiber filter (purchased from Whatman) ), fat-soluble scintillation fluid: PPO, POPOP (purchased from Shanghai Reagent No. 1), toluene (purchased from Sinopharm Chemical Reagent Co., Ltd.), and Tris imported and packaged.

Cells: HEK-293 cells stably expressing 5-HT _1A receptor by gene recombination were cultured in 3-5 cells cultured with DMEM + 10% serum, and cells were harvested with PBS, and the cells were centrifuged at -4 to 3000 rpm. After a minute, discard the supernatant, collect the cells, and store in a -80 degree refrigerator. Resuspend with D ₁ Binding Buffer (pH 7.4) during the experiment.

2. Experimental methods:

The competitive inhibition rate of [ ³ H]8-OH-DPAT binding to 5-HT _1A receptor was determined by coarse screening for each compound at a concentration of 10 umol/L.

The specific experimental data is shown in Table 1.

Table 1: Compounds for D ₂ and D ₃ receptor binding assays and nucleophilicity of the 5-HT _1A receptor (Ki: nmol)

Conclusion: It can be seen from the experimental results in Table 1 that the series of compounds of the present invention have strong nucleophilicity to D ₃ and very weak nucleophilicity to D ₂ , which is nearly 10,000 times different, indicating that the series of compounds of the present invention are D ₂ / The D ₃ receptor is highly selective, avoiding the side effects of D ₂ receptor selection; it also exhibits strong nucleophilicity to the 5-HT _1A receptor and increases the broad spectrum of anti-neuropsychiatric diseases. .

Example 13 In vivo anti-schizophrenia activity test of the cyclohexane derivative of the present invention

1, MK-801 model

(1), MK-801 induced mouse schizophrenia experimental model establishment

100 Sprague-Dawley rats (supplied by Shanghai Lake Experimental Animal Co., Ltd.) were male and were randomly divided into 10 groups according to body weight: blank control group, MK-801 model control group, and cyclohexane derivative C1- of the present invention. Group C8 (corresponding to Compound 1 to Compound 7, Compound 14 of the present invention, respectively) (0.3 mg/kg), and a positive control drug, Kallibazine (prepared according to the method of Patent CN 103130737 A) (0.3 mg/kg). Each animal was placed in a soundproof box for 30 minutes on the day before the experiment, and 30 minutes after the administration of the test substance on the next day, the MK-801 solution at a concentration of 0.3 mg/kg was used for intraperitoneal injection according to the body weight of 5.0 mL/kg of the mouse. An experimental model of mouse schizophrenia was established.

Mode of administration: the cyclohexane derivative group of the present invention and the positive control drug cariprazine group are orally administered, MK-801 The model control group was administered intraperitoneally.

(2), behavioral observation of the market

After the mice were administered MK-801, they were immediately placed in a soundproof box, and the total distance of the mouse's voluntary activities in 2.5 hours was observed.

Improvement rate = (total distance of model control group activity - total distance of administration group activity) / (total distance of model control group activity) × 100%.

(3), statistical methods

All data in

Representation, SPSS 17.0 software statistical package processing, two samples were compared t test and one-way ANOVA, P <0.05 was considered as significant difference.

(4), experimental results

See Table 2 below for the specific results.

Table 2: Effect of a single oral administration of a cyclohexane derivative of the invention on the total distance of the open field movement induced by MK-801 in a mouse model of schizophrenia

The results in Table 2 show that the positive control drug kalidazine group and the cyclohexane derivative compounds 1 to 7 of the present invention all reduced the total activity distance in the rat within 150 minutes as compared with the MK-801 model control group. Where *P<0.05, ****P<0.0001.

The results of this test indicate that:

1 Compared with the blank control group, the movement distance of the MK-801 model control group increased significantly, indicating that MK-801 can cause schizophrenia in mice.

2 Compared with the positive control drug carilazide group of MK-801 model, the positive control drug kalidazine group significantly inhibited MK-801-induced high spontaneous activity in rats at a dose of 0.3 kg/kg (P<0.0001); The cyclohexane derivative compounds 1, 2, 3 and 5 of the present invention significantly inhibited MK-801-induced high spontaneous activity in rats at a dose of 0.3 kg/kg (P < 0.0001), the cyclohexane derivative of the present invention. Compounds 6, 7 and 14 also significantly inhibited MK-801-induced high spontaneous activity in rats at a dose of 0.3 kg/kg (P<0.05), which was comparable to the positive control drug kalidazine group. Since the MK-801-induced open field exercise model is closely related to the symptoms of schizophrenia, it is indicated that the cyclohexane derivative series of the present invention has a strong anti-schizophrenia symptom.

Example 14 Study on Acute Toxicity of Cyclohexane Derivatives of the Invention

The ICR mice were evaluated for oral administration of the compounds of the present invention, and the signs of toxicity and mortality of the animals after oral administration were observed, and the acute toxicity was compared by the Bliss method.

Experimental program

(1) Preparation of solvent: Weigh the appropriate amount of Tween-80 and dilute with deionized water to a concentration of 5% (g/v) Tween-80.

(2), the preparation of the drug: separately weighed the required test article, with 5% Tween 80 solution formulated to a concentration of 0.94, 1.88, 3.75, 7.5, 15, 30, 60mg / mL (equivalent to 18.75, respectively 37.5, 75, 150, 300, 600, 1200 mg/kg) suspension.

(3) Route of administration: The administration routes of the test sample and the vehicle control group (0.5% Tween-80) were all orally administered.

General symptom observation: On the day of administration, the observation was performed once every 10 minutes, 0.5, 2, 4, and 6 hours after the first administration; the observation period was 2-6 days, observed twice a day, once in the morning and afternoon. . Observations include, but are not limited to, general conditions, behavioral activities, gait posture, eyes, mouth, nose, gastrointestinal tract, skin coat, genitourinary tract.

The experimental results are shown in Table 4.

Table 4: Results of acute toxicity test of single oral administration of compound

Conclusion: It shows that the acute toxicity of the cyclohexane derivative of the present invention is far lower than that of the positive control drug callithazine (RGH-188, 75.3 mg/kg), and the safety is good.

Claims (13)

1. A cyclohexane derivative or a stereoisomer or salt thereof, wherein the structure of the cyclohexane derivative is as shown in the following formula IB:

   

   Where X is N or C;

   R is

   

   And, the R group may be optionally substituted by one or more substituents selected from halogen, substituted or unsubstituted C ₁ -C ₆ alkyl; the halogen is selected from F, Cl, One or more of Br or I; the substituted or unsubstituted C ₁ -C ₆ alkyl group is selected from a substituted or unsubstituted C ₁ -C ₄ alkyl group, such as methyl, ethyl, propyl or Butyl, the substituent is a halogen, such as one or more of F, Cl, Br or I, and the substituted C ₁ -C ₄ alkyl group is preferably a trifluoromethyl group.
2. A cyclohexane derivative or a stereoisomer or salt thereof, wherein the cyclohexane derivative has the following structure:

   

   Where R is

   

   And, the R group may be optionally substituted by one or more substituents selected from halogen, substituted or unsubstituted C ₁ -C ₆ alkyl; the halogen is selected from F, Cl, One or more of Br or I; the substituted or unsubstituted C ₁ -C ₆ alkyl group is selected from a substituted or unsubstituted C ₁ -C ₄ alkyl group, such as methyl, ethyl, propyl or Butyl, the substituent is one or more of halogen: F, Cl, Br or I, and the substituted C ₁ -C ₄ alkyl group is preferably a trifluoromethyl group.
3. The cyclohexane derivative or a stereoisomer or salt thereof according to claim 1 or 2, wherein the stereoisomer is a cis stereoisomer or a trans stereoisomer, preferably Trans stereoisomers.
4. The cyclohexane derivative or a stereoisomer or salt thereof according to claim 1 or 2, wherein the salt of the cyclohexane derivative is formed of a cyclohexane derivative and an acid, and the acid is An organic or inorganic acid selected from the group consisting of hydrochloric acid, sulfuric acid, nitric acid or phosphoric acid; said organic acid being selected from the group consisting of formic acid, acetic acid, oxalic acid malonic acid, maleic acid, fumaric acid, succinic acid or benzoic acid; Other physiologically acceptable salts.
5. The cyclohexane derivative or a stereoisomer or salt thereof according to claim 1 or 2, wherein the cyclohexane derivative or a stereoisomer or salt thereof is selected from the following compounds or salts thereof:

   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-fluoro-benzo[d]isoxazole)-3-piperazinyl]ethyl]cyclohexyl]-N,N-dimethyl Urea, compound 5;

   N'-[trans-4-[2-[4-(3-chloro-benzo[d]isoxazole)-6-piperazinyl]ethyl]cyclohexyl]-N,N-dimethyl Urea, compound 6;

   N'-[trans-4-[2-[4-(6-fluoro-benzo[d]isoxazole)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]isothiazole)-3-piperidinyl]ethyl]cyclohexyl]-N,N-dimethylurea, compound 11;

   N'-[trans-4-[2-[4-(3-chloro-benzo[d]isoxazole)-6-piperidinyl]ethyl]cyclohexyl]-N,N-dimethyl Urea, compound 12;

   N'-[trans-4-[2-[4-(3-methyl-benzo[d]isoxazole)-6-piperazinyl]ethyl]cyclohexyl]-N,N-dimethyl Base urea, compound 13;

   N'-[trans-4-[2-[4-(6-methyl-benzo[d]isoxazole)-4-piperazinyl]ethyl]cyclohexyl]-N,N-dimethyl Base urea, compound 14.
6. A method for producing a cyclohexane derivative or a stereoisomer or salt thereof according to claim 1 or 2, wherein the method comprises the steps of:

   4-ethylcyclohexylamine derivative II is reacted with N,N-diformylcarbonyl chloride III in the presence of an acid binding agent to give compound IB:

   

   Wherein R, X are as defined in claim 1 or 2;

   The molar ratio of the compound II to the compound III is 1-1.5:1, the reaction temperature is 0 ° C -50 ° C, the acid binding agent is an organic basic substance or an inorganic basic substance; the organic basic substance is selected from the group consisting of triethylamine, One or more of diisopropylethylamine or pyridine selected from one or more of sodium carbonate, potassium carbonate, sodium hydrogencarbonate or potassium hydrogencarbonate, the acid binding agent The molar ratio to the compound II is 1-1.5:1;

   The preparation method of the compound II includes the following steps:

   

   Wherein R, X are as defined in claim 1 or 2, and Pg is an amino protecting group;

   (1) When X is nitrogen, a piperazine and a bromine are subjected to a coupling reaction to obtain an intermediate IV;

   Wherein the coupling reaction is carried out under the palladium catalysis, in the presence of a strong base potassium t-butoxide, sodium t-butoxide, potassium carbonate or cesium carbonate, the reaction temperature is 50-150 ° C, piperazine and bromine The molar ratio is 1-5:1; or

   When X is C, intermediate IV is commercially available;

   (2) Intermediate IV and 4-amino-protected cyclohexane acetaldehyde are subjected to a condensation imine reduction reaction to obtain an intermediate V;

   Wherein the reducing agent is a boron compound selected from sodium triacetoxyborohydride or sodium borohydride; the molar ratio of the intermediate IV to the 4-amino protected cyclohexane acetaldehyde is 1-1.2:1; The molar ratio of the compound to the intermediate IV is 1-2:1;

   (3) a deamination protecting group;

   When the amino protecting group is t-butoxycarbonyl Boc, an acid deamination protecting group is added; the acid is an organic solution of hydrogen chloride or trifluoroacetic acid;

   When the amino protecting group is benzyl Bn or benzyl formate CBz, a palladium-carbon hydrodeamination protecting group is used, and the hydrogenation pressure is from 0.1 to 1 MPa to obtain a compound of formula II.
7. The use of the cyclohexane derivative or the stereoisomer or salt thereof according to claim 1 or 2 for the preparation of a medicament for anti-neuropsychiatric diseases.
8. The use according to claim 7, wherein the application means that the cyclohexane derivative or a stereoisomer or salt thereof is prepared for improving/treating schizophrenia, mental disorder, mental disorder, confusion, mood disorder, Use in drugs for bipolar disorder, depression, phobia, obsessive-compulsive disorder, anxiety disorder or cognitive disorder.
9. The use according to claim 7 or 8, wherein the drug is composed of a cyclohexane derivative of the formula IB or the formula I or a stereoisomer or salt thereof as an active ingredient and a pharmaceutically acceptable auxiliary. A pharmaceutical composition which is a solid preparation or a liquid preparation for oral, gastrointestinal, buccal, sublingual, nasal, rectal or transdermal administration.
10. The use according to claim 9, wherein the pharmaceutical composition is composed of a cyclohexane derivative of the formula IB or the formula I or a stereoisomer or salt thereof as an active ingredient and a pharmaceutically acceptable auxiliary. a solid tablet; the solid tablet consists of the following weight ratio components:

    

    The diluent is selected from the group consisting of starch, lactose or microcrystalline cellulose; the binder is selected from the group consisting of hydroxybenzylcellulose, hydroxypropylmethylcellulose or polyvinylpyrrolidone; the disintegrant is selected from hydroxyB Based on sodium starch or crospovidone, the lubricant is magnesium stearate.
11. The use according to claim 10, wherein the pharmaceutical composition is N'-[trans-4-[2-[4-(benzo[b]thiophene)-7-piperazinyl] Ethyl]cyclohexyl]-N,N-dimethylurea compound as a solid tablet composed of an active ingredient and a pharmaceutical excipient; the solid tablet is composed of the following weight ratio components:

    

    The diluent is selected from the group consisting of starch, lactose or microcrystalline cellulose, the binder is selected from the group consisting of hydroxybenzylcellulose, hydroxypropylmethylcellulose or polyvinylpyrrolidone, and the disintegrant is selected from sodium hydroxyethyl starch or Cross-linked povidone, the lubricant is magnesium stearate.
12. The use according to claim 9, wherein the composition is composed of a cyclohexane derivative of the formula IB or the formula I or a stereoisomer or salt thereof as an active ingredient and a pharmaceutically acceptable auxiliary. a suspension; the suspension consists of the following weight ratio components:

    

    The suspending agent is selected from the group consisting of xanthan gum or microcrystalline cellulose; the preservative is selected from sodium benzoate, methyl p-hydroxybenzoate or ethyl p-hydroxybenzoate, the diluent being selected from water or sorbitol; The buffering agent is citrate; the co-solvent is selected from the group consisting of cyclodextrin, ethanol, propylene glycol or polyethylene glycol; the flavoring agent is a sweetener selected from sugar or saccharin; the coloring agent is a fat-soluble coloring agent or water-soluble A coloring agent selected from the group consisting of carotene, cocoa pigment, and caramel color.
13. The use according to claim 12, wherein said pharmaceutical composition is composed of N'-[trans -4-[2-[4-(Benzo[b]thiophene)-7-piperazinyl]ethyl]cyclohexyl]-N,N-dimethylurea compound as a mixture of active ingredients and pharmaceutical excipients a suspension; the suspension consists of the following weight ratio components:

    

    The suspending agent is selected from the group consisting of xanthan gum or microcrystalline cellulose, and the preservative is selected from the group consisting of sodium benzoate, methyl p-hydroxybenzoate or ethyl p-hydroxybenzoate, the diluent is selected from the group consisting of water, sorbitol, and the buffer is lemon. The acid salt, the co-solvent is selected from the group consisting of cyclodextrin, ethanol, propylene glycol or polyethylene glycol, the flavoring agent is a sweetener selected from sugar or saccharin; the coloring agent is a fat-soluble coloring agent or a water-soluble coloring agent selected from the group consisting of carotene , cocoa pigment or caramel color.