CN1227550A - Troponyl piperazines as selective dopamine D4 receptor ligands - Google Patents

Abstract

The present invention discloses compounds of general formula i, stereoisomers, solvates, or pharmaceutically acceptable salts thereof, wherein: r₁Is H, halo, aryl or substituted by one or two independently selected from halo, C_1－4Alkyl radical, C_1－4Aryl substituted with alkoxy, nitro, trifluoromethyl, trifluoromethoxy or cyano; r₂Is C_4－9An alkyl group; phenyl or substituted by one or two groups independently selected from OH, cyano, halo, C_1－4Alkoxy radical, C_1－4Phenyl substituted with alkyl, trifluoromethyl, trifluoromethoxy, nitro or phenyl groups; a naphthyl group; phenyl fused with a 5 or 6 membered heterocyclic ring, a coumarin moiety wherein R is₃And R₄Independently selected from H, C_1－4Alkyl and C_1－4An alkoxy group; or 1, 2-methylenedioxyphenyl; with the proviso that when R₁When is H, R₂Is not phenyl or 3, 4-dimethoxyphenyl. It is also disclosed for use as a medicament for the treatment of indications in which the dopamine D4 receptor is involved, such as schizophrenia and anxiety.

Description

Tropotrienylpiperazines as selective dopamine D4 receptor ligands

The present invention relates to a compound that binds to dopamine D4 receptor, its preparation method and its application in treatment and drug screening.

Background of the invention

Neuronal cell receptors that bind the neurotransmitter dopamine form a group of at least five structurally distinct proteins that can now be produced using recombinant DNA techniques. These techniques have been applied to construct cell lines that incorporate dopamine receptors in their membranes to provide a reproducible and uniform substrate with which chemical libraries can be screened to identify potential CNS-active drugs.

Recent evidence significantly demonstrates that dopamine receptors are classified as D4 in the etiology of schizophrenia. Compounds capable of interfering with the function of this receptor, which is normally present in six times greater quantities in schizophrenia, have been shown to be useful in the treatment of this disorder (Seeman et al., Nature, 1993, 365:441). Indeed, some drugs currently on the market show antagonism of the desired D4 receptor activity and bind this receptor with relatively strong affinity. However, due to their structure, these drugs also interact with related dopamine receptors, especially the D2 receptor type, which leads to significant side effects including altered motor function and tachycardia. There is therefore a need to provide compounds which not only exhibit a high affinity for the D4 receptor but also a relatively low affinity for the D2 receptor. In this specification, this combination of desirable receptor binding properties is referred to as D4 selectivity.

Currently marketed products for the treatment of diseases involving D4 receptor function include dibenzodiazepine, clozapine, and dibenzoxazepine, isoloxapine. Analysis of the dopamine receptor binding properties of both products showed approximately 10-fold preferential binding to D4 receptors over D2 receptors. Similarly, both products bind the D4 receptor with approximately the same affinity (Ki value ~20 nM). Other products recently published in the scientific literature showed similar D4-D2 selectivity patterns and D4 affinity values.

The use of certain tropotrienylpiperazines as dopamine receptor antagonists, for example in the treatment of Parkinson's syndrome and other related diseases, is disclosed in EP 034,894 and related scientific articles (see Bagli, J. et al., J. Med. Chem., 1984, 27:875 and Bagli, J. et al., (J. Med. Chem., 1986, 29:186).

The object of the present invention is to provide compounds that bind to the D4 receptor.

The object of the present invention is to provide D4 receptor binding compounds.

Another object of the present invention is to provide compounds which bind selectively to D4 receptors, in particular relative to D2 receptors.

A further object of the present invention is to provide pharmaceutical compositions comprising the compounds of the present invention as active ingredients.

Another object of the present invention is to provide methods of effectively treating medical conditions such as schizophrenia and anxiety for which administration of D4 receptor antagonists is appropriate.

Summary of the invention

One aspect of the present invention provides a compound of formula I, its stereoisomer, solvate, or pharmaceutically acceptable salt:

Wherein _R is H, halo, aryl or is independently selected from one or two halo, C _1-4 alkyl, C _1-4 alkoxy, nitro, trifluoromethyl, trifluoromethoxy or An aryl group substituted with a cyano group;

R ₂ is C _4-9 alkyl; phenyl or one or two independently selected from OH, cyano, halogen, C _1-4 alkoxy, C _1-4 alkyl, trifluoromethyl, trifluoro Phenyl substituted with radicals of methoxy, nitro or phenyl; naphthyl; phenyl fused to a 5- or 6-membered heterocycle, coumarin moiety

Wherein R ₃ and R ₄ are independently selected from H, C _1-4 alkyl and C _1-4 alkoxy; or 1,2-methylenedioxyphenyl;

with the proviso that when _R1 is H, _R2 is not phenyl or 3,4-dimethoxyphenyl.

Another aspect of the present invention provides a pharmaceutical composition comprising an effective amount of compounds of formulas I and II and a pharmaceutically acceptable carrier for antagonizing D4 receptor stimulation;

wherein _{R is} independently selected from H, halogen, aryl or is independently selected from H, halogen, C _1-4 alkyl, C _1-4 alkoxy, nitro, trifluoromethyl, tri Aryl substituted with fluoromethoxy or cyano groups;

R ₅ is independently selected from C _4-9 alkyl; phenyl is selected from one or two independently selected from OH, cyano, halogen, C _1-4 alkoxy, C _1-4 alkyl, trifluoromethyl , trifluoromethoxy, nitro or phenyl group substituted phenyl; naphthyl; phenyl fused to a 5- or 6-membered heterocycle, coumarin moiety

Wherein R ₃ and R ₄ are independently selected from H, C _1-4 alkyl and C _1-4 alkoxy; or 1,2-methylenedioxyphenyl.

This is essentially Formula I without conditions. Formula II is used as a pharmaceutical composition.

Another aspect of the present invention provides the compounds of formulas I and II as D4 receptor antagonists in the treatment of medical symptoms mediated by inhibition of D4 receptor antagonism, and in the treatment of schizophrenia and anxiety.

The invention further provides assays in which the compounds of the invention are used in a population of receptors to distinguish D4 receptors from other types of receptors, especially D2 receptors. These and other aspects of the invention are described in more detail below.

Detailed description and preferred embodiments

definition:

The term "C _4-9 alkyl" as used herein refers to straight-chain alkyl groups containing 4-9 carbon atoms and branched-chain alkyl groups containing 6-8 carbon atoms and includes pentyl, hexyl, 1-methylhexyl, etc. wait.

The term "C _1-4 alkyl" as used herein refers to a straight chain alkyl group containing 1-4 carbon atoms and a branched chain alkyl group containing 3-4 carbon atoms and includes methyl, ethyl, propyl, 1- Methylethyl, butyl, 1-methylpropyl, etc. 1-Methylethyl and 1-methylpropyl are also known as isopropyl and sec-butyl, respectively.

The term "C _1-4 alkoxy" as used herein refers to straight-chain alkoxy containing 1-4 carbon atoms and branched-chain alkoxy containing 3-4 carbon atoms and includes methoxy, ethoxy, 1-methylethoxy, propoxy, butoxy, etc.

The term "halo" as used herein refers to halides and includes fluorine, chlorine, bromine and iodine.

The term "heterocycle" as used herein refers to a 5- or 6-membered saturated or unsaturated ring, wherein the heterocycle contains 1-3 heteroatoms independently selected from nitrogen, oxygen and sulfur, and includes 1,3-dioxane Pentene, thiophene, furan, pyrrole, imidazole, pyrazole, pyrrolidine, pyrroline, imidazolidine, pyrazolidine, pyrazoline, pyran, pyridine, pyrazine, pyrimidine, pyridazine, piperidine, piperazine, Morpholine etc. Thus, some examples of "phenyl fused to a 5- or 6-membered heterocycle" are benzothiophene, isobenzofuran, benzopyran, indole, isoindole, indazole, isoquinoline, quinoline , Phthalazine, quinoxaline, quinazoline, o-phthalazine, isochroman, chroman, indoline, isoindoline, etc.

The term "aryl" as used herein refers to a 5- or 6-membered unsaturated carbocyclic ring system, which can be either an aromatic ring (such as phenyl) or a ring containing one or two nitrogen atoms, one or two oxygen atoms, one nitrogen atom and an oxygen atom, a nitrogen atom and a sulfur atom, or a heteroaromatic ring of a sulfur atom, including but not limited to thiophene, furan, pyrrole, pyran, pyridine, pyrazine, pyrimidine, pyridazine, isothiazole, isoxa azoles, pyrazolines, and pyrazines. These aryl groups can be selected by one or two independently selected from halogen, C _1-4 alkyl, C _1-4 alkoxy, nitro, trifluoromethyl, trifluoromethane Oxygen and cyano groups are substituted. A preferred aryl group is phenyl substituted with a substituent in the meta or para position.

The term "pharmaceutically acceptable salt" refers to an acid addition salt or a base addition salt that is compatible with the treatment of the patient for the intended use.

"Pharmaceutically acceptable acid addition salt" is any non-toxic organic or inorganic acid addition salt of a basic compound represented by formula I or any intermediate thereof. Examples of inorganic acids which form suitable salts include hydrochloric, hydrobromic, sulfuric and phosphoric acids and acid metal salts such as sodium monohydrogen orthophosphate and potassium hydrogensulfate. Examples of organic acids which form suitable salts include mono-, di- and tricarboxylic acids. Examples of such acids include, for example, acetic acid, glycolic acid, lactic acid, pyruvic acid, malonic acid, succinic acid, glutaric acid, fumaric acid, malic acid, tartaric acid, citric acid, ascorbic acid, maleic acid, hydroxymaleic acid, benzoic acid, hydroxybenzoic acid, phenylacetic acid, cinnamic acid, salicylic acid, 2-phenoxybenzoic acid, p-toluenesulfonic acid and sulfonic acids such as methanesulfonic acid and 2-hydroxyethanesulfonic acid. Mono- or di-acid salts may be formed and such salts may exist in hydrated, solvated or substantially anhydrous form. In general, the acid addition salts of these compounds are readily soluble in water and various hydrophilic organic solvents, and generally have higher melting points than their free base forms.

"Pharmaceutically acceptable base addition salt" means a non-toxic organic or inorganic base addition salt of a compound of formula (I) or any intermediate thereof. Examples are alkali metal or alkaline earth metal hydroxides such as sodium hydroxide, potassium hydroxide, calcium hydroxide, magnesium hydroxide or barium hydroxide; ammonia, and aliphatic, cycloaliphatic or aromatic organic amines such as methylamine, trimethylamine amines, and picolines. It is important to choose an appropriate salt so that its ester is not hydrolyzed. Such criteria for selecting appropriate salts are known to those skilled in the art.

"Solvate" refers to a compound of formula I or a pharmaceutically acceptable salt of formula I incorporating appropriate solvent molecules in the crystal lattice. Suitable solvents are those in which the solvate is administered in substantially nontoxic amounts to achieve the desired effect. Examples of suitable solvents are ethanol and the like.

The term "stereoisomers" refers to a general term for all isomers of a single molecule that differ only in the orientation of the atoms in space. It includes mirror image isomers (enantiomers), geometric (cis/trans) isomers, and isomers of compounds with more than one chiral center that are not mirror images of each other (diastereoisomers) .

The term "patient" refers to warm-blooded animals such as rats, mice, dogs, cats, guinea pigs, and primates such as humans.

The term "treating" refers to alleviating symptoms, temporarily or permanently eliminating the cause of symptoms, preventing or slowing the onset of symptoms of the disease or symptoms.

The term "therapeutically effective amount" refers to the amount of a compound effective to treat the disease or condition in question.

The term "pharmaceutically acceptable carrier" refers to a non-toxic solvent, dispersant, excipient, adjuvant or other substance with which the active ingredient is mixed in order to form a pharmaceutical composition, ie, a dosage form that can be administered to a patient. An example of such a carrier is a pharmaceutically acceptable oil commonly used for parenteral administration.

The term "schizophrenia" refers to schizophrenia, schizophrenia-like disorders, schizoaffective disorders and psychosis, wherein the term "psychotic" involves delusions, marked hallucinations, incoherent or disorganized or catatonic behavior. See Diagnostic and Statistical Manual of Mental Disorder, fourth edition, American Psychiatric Association, Washington D.C.

The present invention relates to compounds that bind to dopamine D4 receptors in a selective manner relative to dopamine D2 receptors.

In an embodiment of the invention, compounds of formulas I and II include those compounds wherein R ₁ is H and R ₂ is C _4-9 alkyl.

In a particular embodiment of the invention, in the compounds of formulas I and II, R ₁ is H and R ₂ is n-pentyl.

In another embodiment of the present invention, the compounds of formulas I and II include wherein _R1 is H and _R2 is selected from naphthyl, phenyl, one or two selected from methyl, methoxy, nitro, bromo, Those compounds of phenyl substituted with groups of chlorine, fluorine, trifluoromethoxy, trifluoromethyl, phenyl and cyano and phenyl fused to a 5- or 6-membered heterocycle.

In a particular embodiment of the invention, compounds of formulas I and II include those compounds wherein R _is H and R _is :

4-methylphenyl;

4-trifluoromethoxyphenyl;

4-trifluoromethylphenyl;

4-nitrophenyl;

3-cyanophenyl;

3-trifluoromethylphenyl;

4-bromophenyl;

3,5-difluorophenyl;

2-cyanophenyl;

2-chlorophenyl;

2-nitrophenyl;

3-methylphenyl;

2-phenylphenyl;

3-bromophenyl;

3,5-bis(trifluoromethyl)phenyl;

3-nitrophenyl;

4-cyanophenyl;

3-methoxyphenyl;

2-naphthyl;

1,2-methylenedioxyphenyl; (preferably, _R forms piperonyl with the methylene it is attached to)

4-Chlorophenyl; and

3-Chlorophenyl.

In another embodiment of the present invention, the compound of formula I and II is wherein R ₁ is H and R ₂ is a coumarin moiety as shown below, R ₃ and R ₄ are selected from H and C _1-4 alkoxy those compounds.

A particular embodiment of the invention includes compounds of formulas I and II wherein R _is H and _R is 1-(6,7-dimethoxycoumarin) and 1-(7-methoxycoumarin) .

In another embodiment of the present invention, the compounds of formula I and II include wherein R ₁ is selected from halogen, aryl, and one or two are independently selected from H, halogen, C _1-4 alkyl, C _1-4 Alkoxy, nitro, trifluoromethyl, trifluoromethoxy and cyano groups substituted aryl, _R2 is selected from C _4-9 alkyl, naphthyl, phenyl, by one or two Phenyl substituted with a group selected from methyl, methoxy, nitro, bromo, chloro, fluoro, trifluoromethoxy, trifluoromethyl, cyano and phenyl, fused to a 5- or 6-membered heterocyclic ring Combined phenyl and coumarin moieties as shown below, wherein _R3 and _R4 are selected from H and _C1-4 alkoxy.

In a more preferred embodiment of the present invention, compounds of formulas I and II include wherein R ^is selected from halogen and aryl attached at carbon 7 of a 7-membered ring and R is selected from one or _two of methyl, methoxy, nitro, bromo, chloro, fluoro, trifluoromethoxy, trifluoromethyl, cyano and phenyl substituted with groups of phenyl; or _R2 is naphthyl or hetero with 5 or 6 members Those compounds with ring-fused phenyl groups.

In particular embodiments of the present invention, compounds of formula I and II include wherein R ₁ is selected from bromine and thienyl attached to carbon 7 of the 7-membered ring and R ₂ is selected from 1,2-methylenedioxyphenyl and those compounds of phenyl substituted by chlorine.

In particular embodiments of the invention, compounds of formulas I and II include:

2-(4-Benzyl-1-piperazinyl)-2,4,6-cycloheptatrien-1-one;

2-(4-Piperonyl-1-piperazinyl)-2,4,6-cycloheptatrien-1-one;

2-(4-Hexyl-1-piperazinyl)-2,4,6-cycloheptatrien-1-one;

2-[4-(3-methoxybenzyl)-1-piperazinyl]-2,4,6-cycloheptatrien-1-one;

2-[4-(4-cyanobenzyl)-1-piperazinyl]-2,4,6-cycloheptatrien-1-one;

2-[4-(4-Methylbenzyl)-1-piperazinyl]-2,4,6-cycloheptatrien-1-one;

2-[4-(4-Trifluoromethoxybenzyl)-1-piperazinyl]-2,4,6-cycloheptatrien-1-one;

2-[4-(4-Trifluoromethylbenzyl)-1-piperazinyl]-2,4,6-cycloheptatrien-1-one;

2-[4-(4-nitrobenzyl)-1-piperazinyl]-2,4,6-cycloheptatrien-1-one;

2-[4-(3-cyanobenzyl)-1-piperazinyl]-2,4,6-cycloheptatrien-1-one;

2-[4-(3-Trifluoromethylbenzyl)-1-piperazinyl]-2,4,6-cycloheptatrien-1-one;

2-[4-(4-Bromobenzyl)-1-piperazinyl]-2,4,6-cycloheptatrien-1-one;

2-[4-(4-naphthyl-2-methyl)-1-piperazinyl]-2,4,6-cycloheptatrien-1-one;

2-[4-(3,5-difluorobenzyl)-1-piperazinyl]-2,4,6-cycloheptatrien-1-one;

2-[4-(2-Chlorobenzyl)-1-piperazinyl]-2,4,6-cycloheptatrien-1-one;

2-[4-(3-Methylbenzyl)-1-piperazinyl]-2,4,6-cycloheptatrien-1-one;

2-[4-(2-Phenylbenzyl)-1-piperazinyl]-2,4,6-cycloheptatrien-1-one;

2-[4-(3-Bromobenzyl)-1-piperazinyl]-2,4,6-cycloheptatrien-1-one;

2-{4-[3,5-bis(trifluoromethyl)benzyl]-1-piperazinyl}-2,4,6-cycloheptatrien-1-one;

2-[4-(3-nitrobenzyl)-1-piperazinyl]-2,4,6-cycloheptatrien-1-one;

2-[4-(3-Chlorobenzyl)-1-piperazinyl]-2,4,6-cycloheptatrien-1-one;

2-[4-(4-Chlorobenzyl)-1-piperazinyl]-2,4,6-cycloheptatrien-1-one;

7-Bromo-2-[4-(4-chlorobenzyl)-1-piperazinyl]-2,4,6-cycloheptatrien-1-one;

7-bromo-2-(4-piperonyl-1-piperazinyl)-2,4,6-cycloheptatrien-1-one;

2-(4-Piperonyl-1-piperazinyl)-7-thiophen-2-yl-2,4,6-cycloheptatrien-1-one;

2-{4-[1-(6,7-dimethoxycoumarin)methyl]-1-piperazinyl}-2,4,6-cycloheptatrien-1-one; and

2-{4-[1-(7-Methoxycoumarin)methyl]-1-piperazinyl}-2,4,6-cycloheptatrien-1-one.

Preferred compounds of formulas I and II which enhance the potency of D4 are as follows:

2-(4-Piperonyl-1-piperazinyl)-2,4,6-cycloheptatrien-1-one;

2-(4-Hexyl-1-piperazinyl)-2,4,6-cycloheptatrien-1-one;

2-[4-(3-methoxybenzyl)-1-piperazinyl]-2,4,6-cycloheptatrien-1-one;

2-[4-(4-cyanobenzyl)-1-piperazinyl]-2,4,6-cycloheptatrien-1-one;

2-[4-(4-Methylbenzyl)-1-piperazinyl]-2,4,6-cycloheptatrien-1-one;

2-[4-(4-Trifluoromethoxybenzyl)-1-piperazinyl]-2,4,6-cycloheptatrien-1-one;

2-[4-(4-Trifluoromethylbenzyl)-1-piperazinyl]-2,4,6-cycloheptatrien-1-one;

2-[4-(4-nitrobenzyl)-1-piperazinyl]-2,4,6-cycloheptatrien-1-one;

2-[4-(3-Trifluoromethylbenzyl)-1-piperazinyl]-2,4,6-cycloheptatrien-1-one;

2-[4-(4-Bromobenzyl)-1-piperazinyl]-2,4,6-cycloheptatrien-1-one;

2-[4-(4-naphthyl-2-methyl)-1-piperazinyl]-2,4,6-cycloheptatrien-1-one;

2-[4-(3-Bromobenzyl)-1-piperazinyl]-2,4,6-cycloheptatrien-1-one;

2-[4-(3-Chlorobenzyl)-1-piperazinyl]-2,4,6-cycloheptatrien-1-one;

2-[4-(4-Chlorobenzyl)-1-piperazinyl]-2,4,6-cycloheptatrien-1-one;

7-Bromo-2-(4-piperonyl-1-piperazinyl)-2,4,6-cycloheptatrien-1-one; and

2-(4-Piperonyl-1-piperazinyl)-7-thiophen-2-yl-2,4,6-cycloheptatrien-1-one.

Particularly preferred compounds of formulas I and II include:

2-(4-Piperonyl-1-piperazinyl)-2,4,6-cycloheptatrien-1-one;

2-[4-(3-methoxybenzyl)-1-piperazinyl]-2,4,6-cycloheptatrien-1-one;

2-[4-(4-cyanobenzyl)-1-piperazinyl]-2,4,6-cycloheptatrien-1-one;

2-[4-(4-Bromobenzyl)-1-piperazinyl]-2,4,6-cycloheptatrien-1-one;

2-[4-(4-Trifluoromethoxybenzyl)-1-piperazinyl]-2,4,6-cycloheptatrien-1-one;

2-[4-(4-Trifluoromethylbenzyl)-1-piperazinyl]-2,4,6-cycloheptatrien-1-one;

2-[4-(4-nitrobenzyl)-1-piperazinyl]-2,4,6-cycloheptatrien-1-one;

2-[4-(3-Chlorobenzyl)-1-piperazinyl]-2,4,6-cycloheptatrien-1-one;

2-[4-(4-Chlorobenzyl)-1-piperazinyl]-2,4,6-cycloheptatrien-1-one;

2-[4-(4-Methylbenzyl)-1-piperazinyl]-2,4,6-cycloheptatrien-1-one;

7-Bromo-2-(4-piperonyl-1-piperazinyl)-2,4,6-cycloheptatrien-1-one; and

2-[4-(4-Naphthyl-2-methyl)-1-piperazinyl]-2,4,6-cycloheptatrien-1-one.

Most preferred compounds of formulas I and II include:

2-(4-Piperonyl-1-piperazinyl)-2,4,6-cycloheptatrien-1-one;

2-[4-(4-Bromobenzyl)-1-piperazinyl]-2,4,6-cycloheptatrien-1-one;

2-[4-(4-Trifluoromethylbenzyl)-1-piperazinyl]-2,4,6-cycloheptatrien-1-one;

2-[4-(4-Trifluoromethoxybenzyl)-1-piperazinyl]-2,4,6-cycloheptatrien-1-one;

2-[4-(4-nitrobenzyl)-1-piperazinyl]-2,4,6-cycloheptatrien-1-one;

2-[4-(4-Chlorobenzyl)-1-piperazinyl]-2,4,6-cycloheptatrien-1-one;

7-Bromo-2-(4-piperonyl-1-piperazinyl)-2,4,6-cycloheptatrien-1-one; and

2-[4-(4-Naphthyl-2-methyl)-1-piperazinyl]-2,4,6-cycloheptatrien-1-one.

The acid addition salts of the compounds of formulas I and II are most suitably formed with pharmaceutically acceptable acids and include, for example, formation with inorganic acids such as hydrochloric acid, sulfuric acid or phosphoric acid and organic acids such as succinic acid, maleic acid, acetic acid or fumaric acid acid addition salts. Other non-pharmaceutically acceptable salts such as oxalates may be used eg in the isolation of compounds of formula I for laboratory use, or for subsequent conversion into pharmaceutically acceptable acid addition salts. Also included within the scope of the present invention are the solvates and hydrates of the present invention.

Salts of a given compound are converted to salts of the desired compound by using standard techniques, wherein an aqueous solution of the given salt is treated with a base, such as sodium carbonate or sodium hydroxide solution to liberate the free base, which is then treated with a suitable solvent, such as Ether extraction. The free base is separated from the aqueous phase, dried and treated with the desired acid to give the desired salt.

与式B的试剂：如EP 034,894所述在回流甲醇中偶合并且该方法可以在相应的试剂B通过合成或者商业方式得到时使用。The compounds of the present invention can be prepared by methods analogous to those known in the art. Accordingly, another aspect of the present invention provides a process for the preparation of compounds of formulas I and II, or salts thereof, solvates or hydrates thereof, wherein R ₁ is H or halo, which process comprises one of the following two possible processes. The first method involves a reagent of formula A

With a reagent of formula B: Coupling in refluxing methanol is described in EP 034,894 and this method can be used where the corresponding reagent B is available synthetically or commercially.

Reagents (A) in which R ^is H or halo can be synthesized by known techniques in which the appropriate tropotrienone is treated with a methylating reagent (when ^R is H, see Pietra, F. Chemical Reviews (Chem. Rev.), 1973, 73:293 and when R ¹ is halogen see Takeshita, H.; Mori, A. Synthesis, 1986, 578). For example, tropotrienone is treated with acetone of dimethyl sulfate and potassium carbonate.

The compounds of Reagent B are known, commercially available or can be prepared by conventional methods. For example, a useful method for preparing Reagent B compounds is described in EP 034,894.

与式D的试剂：

X=Cl,Br如本文所述在碱如碳酸钾的存在下在惰性有机溶剂如乙腈中偶合制备。其中R₁是H和卤的试剂C化合物用已知技术合成，例如根据Bagli所述方法(《药物化学杂志》(J.Med.Chem.),1984,27:875)用哌嗪处理试剂A。试剂D可以从商业上获得或者可以用已知技术合成，例如用卤化试剂如CBr₄和三苯膦(X=Br)或亚硫酰氯(X=Cl)处理相应的醇类。Compounds of formulas I and II can also be passed through reagents of formula C:

With a reagent of formula D:

X=Cl,Br is prepared by coupling as described herein in the presence of a base such as potassium carbonate in an inert organic solvent such as acetonitrile. Compounds of reagent C, wherein _R is H and halo, are synthesized by known techniques, such as treatment of reagent A with piperazine according to the method described by Bagli (J. Med. Chem., 1984, 27:875). . Reagent D can be obtained commercially or can be synthesized by known techniques, such as treatment of the corresponding alcohol with a halogenating reagent such as _CBr4 and triphenylphosphine (X=Br) or thionyl chloride (X=Cl).

wherein _R is aryl or is replaced by one or two independently selected from H, halo, C _1-4 alkyl, C _1-4 alkoxy, nitro, trifluoromethyl, trifluoromethoxy and cyano Compounds of formulas I and II with aryl substituted by radical groups can be obtained from compounds of formulas I and II wherein R is _halo using standard palladium-catalyzed cross-coupling techniques with metal-aryl compounds _R1 suitable for cross-coupling reactions. -M ₁ (wherein R ₁ is as defined above, and M is an optionally substituted metal substituent attached to any carbon junction of the aryl group) is prepared by reaction. Examples of such M groups include (alkyl) ₃ Sn-, (alkyl) ₂ B-, (HO) ₂ B-, (alkoxy) ₂ B-, Li, Cu, Zn chloride or Mg halide. The most preferred M group is Zn chloride. The reaction is carried out in an inert solvent, and optionally in the presence of a base, lithium chloride and a suitable catalyst. Suitable catalysts include palladium(II) and palladium(0) species such as palladium(II) acetate, palladium(II) chloride and tetrakis(triphenylphosphine)palladium(0). A preferred catalyst is tetrakis(triphenylphosphine)palladium(0). Suitable inert solvents include acetonitrile, N,N-dimethylformamide and tetrahydrofuran, tetrahydrofuran being preferred. The reaction is carried out at a temperature of 25-100°C, preferably 50-100°C. Compounds of formula _R1 -M can be prepared independently or in situ from reagents _R1 -Y wherein Y is a halo or triflate, using standard metallation reactions. For example, compound _R1 -M wherein M is chloro-Zn can be prepared in situ by treating _R1 -Br with n-butyllithium and zinc chloride in tetrahydrofuran at -78°C prior to the cross-coupling reaction.

The clozapine-like binding mode of the compound of the present invention indicates that it can be used as a nerve inhibitor to treat various diseases involving D4 receptor stimulation, such as the drug application for treating anxiety and schizophrenia. Therefore, another aspect of the present invention provides a pharmaceutical composition for the treatment of D4-related medical conditions, wherein the compound of formula II is present in an amount effective to antagonize D4 receptor stimulation, together with a pharmaceutically acceptable carrier. Compounds of formula II are those encompassed by formula I, but also include compounds wherein _R2 is phenyl or 3,4-dimethoxyphenyl. Another aspect of the present invention provides a method of treating a medical condition for which a D4 antagonist is required, the method comprising the step of administering to a patient a compound of formula II in an amount effective to antagonize D4 receptor stimulation together with a pharmaceutically acceptable carrier.

For pharmaceutical use, the compounds of the invention can be administered in the form of standard pharmaceutical compositions. Therefore, another aspect of the present invention provides a pharmaceutical composition comprising a pharmaceutically acceptable carrier and an effective amount of a compound of formula II or a pharmaceutically acceptable salt, solvate or hydrate thereof for antagonizing D4 receptor stimulation.

The compounds of the present invention can be administered by conventional routes, such as oral, parenteral, buccal, sublingual, intranasal, rectal or transdermal administration, and correspondingly prepared pharmaceutical compositions.

When taken orally, the compounds of formulas I and II and their stereoisomers, solvates, hydrates, or pharmaceutically acceptable salts as active ingredients may be formulated into formulations, such as liquid formulations, such as syrups, suspensions or emulsions, or Solid forms such as tablets, capsules, and lozenges. Liquid formulations usually consist of a suspension or solution of the compound or a pharmaceutically acceptable salt thereof in a suitable pharmaceutically acceptable carrier, such as ethanol, glycerol, non-aqueous solvents such as polyethylene glycol, oils, or water, and suspending agents, preservative agent, flavoring or coloring agent. Compositions in tablet form can be prepared with suitable pharmaceutical carriers conventionally used in the preparation of solid formulations. Examples of such carriers include magnesium stearate, starch, lactose, sucrose and cellulose. Compositions in the form of capsules can be prepared by conventional encapsulation methods. For example, pellets containing the active ingredient can be prepared using standard carriers and filled into hard gelatin capsules; or a dispersion or suspension can be prepared in any suitable pharmaceutical carrier, such as aqueous gels, celluloses, silicates or oils, and packed into The dispersion or suspension is filled into soft gelatin capsules.

Typical parenteral compositions consist of a solution or suspension of the compound or a pharmaceutically acceptable salt thereof in a sterile aqueous vehicle or in a parenterally acceptable oil such as polyethylene glycol, polyvinylpyrrolidone, lecithin, peanut oil or sesame oil liquid composition. Alternatively, the solutions can be lyophilized and then reconstituted with an appropriate solvent prior to administration.

Compositions for intranasal administration may conveniently be formulated as aerosols, drops, gels and powders. Aerosol formulations generally consist of a solution or fine suspension of the active substance in a physiologically acceptable aqueous or non-aqueous solvent and are usually presented in single-dose or multi-dose sterile form in a hermetically sealed container, which may be a cartridge or refill. Primed for use with nebulizers. Alternatively, the sealed container is a single dispensing device such as a single dose intranasal inhaler or a single use aerosol dispenser with a metered dose valve. If such dosage forms include an aerosol dispenser, it contains a propellant which may be a compressed gas, such as compressed air, or an organic propellant, such as chlorofluorocarbons. Such aerosol dosage forms may also be in the form of a pump-atomizer.

Compositions suitable for buccal or sublingual administration include tablets, lozenges, and pastilles, wherein the active ingredient is formulated with a carrier such as sugar, acacia, tragacanth, or gelatin and glycerin. Compositions for rectal administration are conveniently presented as suppositories containing conventional suppository bases such as cocoa butter.

Such compositions are preferably in unit dosage form such as tablets, capsules or ampoules. Appropriate unit dosages, ie, therapeutically effective amounts, can be determined during the course of clinical trials appropriately designed for each disease requiring the use of the selected compound, and such dosages will of course vary according to the desired clinical endpoints. Dosage sizes suitable for administration of the compounds of the examples are expected to be generally about the same, or slightly smaller, than those typically used for clozapine. Thus, each dosage unit for oral administration may contain from 1 to 500 mg of active compound and be administered at appropriate frequencies during the initiation and maintenance of treatment.

For use as ligands in the laboratory, the compounds of the invention can be stored in packaged form for reconstitution and use. The compounds of the present invention can be used to distinguish dopamine receptors from other receptor types within the receptor population, such as glutamate and opioid receptors, in particular D4 and D2 receptors. The latter can be accomplished by incubating D4 receptor and D2 receptor preparations with a D4 selective compound of the invention and then incubating the resulting preparation with a radiolabeled dopamine receptor ligand, such as 3H-spirulina. The D2 and D4 receptors were then distinguished by measuring the difference in membrane-bound radioactivity, the D4 receptor showing less radioactivity, ie weaker binding of ³ H-heliperidine.

In another embodiment of the invention, the compound is provided in labeled form, such as radiolabeled form, for example by incorporation ^{of3H or14C} in its structure or by incorporation ^{of125I or123I} . This radiolabeled form can be used directly to distinguish between dopamine D4 and dopamine D2 receptors. Furthermore, radiolabeled forms of the compounds of the invention can be explored to screen for more potent dopamine D4 ligands by determining the ability of the test ligand to displace the radiolabeled compound of the invention.

Example 1 (a) 2-(4-piperonyl-1-piperazinyl)-2,4,6-cycloheptatriene-1-one

2-Methoxytropone (1.36g, 10.0mmol) (Pietra, F. "Chem.Review", 1973,73:293) and 1-piperonylpiperazine (6.61g, 30.0 mmol) in methanol (50.0 mL) was refluxed for 16 hours. After cooling to room temperature, the solvent was removed in vacuo. The residue was subjected to column chromatography using ethyl acetate as eluent. The title compound (1.24 g, 38%) was obtained as a yellow oil. MS(FAB)325(M ⁺ 1).

In a similar manner, the following other compounds were prepared:

(b) Preparation of 7-bromo-2-(4-piperonyl from 7-bromo-2-methoxytropone (Takeshita, H. Mori, A. Synthesis, 1986, 578) -1-piperazinyl)-2,4,6-cycloheptatrien-1-one.

Example 2 (a) 2-(1-piperazinyl)-2,4,6-cycloheptatrien-1-one acetate

This compound was prepared from 2-methoxytropone and piperazine according to the method of Bagli, J et al., J. Med. Chem., 1984, 27:875.

In a similar manner, the following other compounds were prepared:

(b) 7-bromo-2-(1-piperazinyl)-2,4,6-cycloheptatrien-1-one acetate.

Example 3 (a) 2-[4-(3-methoxybenzyl)-1-piperazinyl]-2,4,6-cycloheptatriene-1-one

2-(1-piperazinyl)-2,4,6-cycloheptatrien-1-one acetate (0.171g, 0.680mmol, Example 2a), 3-methoxybenzyl chloride (0.118 g, 0.750 mmol) and potassium carbonate (0.276 g, 2.00 mmol) in acetonitrile (2.00 mL) was stirred at room temperature for 16 hours. Solvent was removed in vacuo. The residue was triturated with dichloromethane and filtered. The filtrate was concentrated to dryness in vacuo and the residue was subjected to preparative thin layer chromatography using ethyl acetate:methanol (9:1) as eluent. The title compound was obtained as a yellow oil (0.095 g, 45%); MS (FAB) 311 (M ⁺ +1).

In a similar manner, the following other compounds were prepared:

b) Preparation of 2-(4-hexyl-1-piperazinyl)-2,4,6-cycloheptatrien-1-one from 1-bromohexane, yellow oil (51%); MS(ES) 275 (M ⁺⁺ 1).

c) Preparation of 2-[4-(4-cyanobenzyl)-1-piperazinyl]-2,4,6-cycloheptatrien-1-one from 4-cyanobenzyl bromide, yellow solid ( 44%); mp 136-138°C; MS (FAB) 306 (M ⁺ +1).

d) Preparation of 2-[4-benzyl-1-piperazinyl]-2,4,6-cycloheptatrien-1-one from benzyl bromide.

e) Preparation of 2-[4-(4-methylbenzyl)-1-piperazinyl]-2,4,6-cycloheptatriene-1-one from α-bromo-p-xylene; MS( FAB) 295 (M ⁺⁺ 1).

f) Preparation of 2-[4-(4-trifluoromethoxybenzyl)-1-piperazinyl]-2,4,6-cycloheptatriene from 4-(trifluoromethoxy)benzyl bromide -1-one; MS (FAB) 365 (M ⁺ +1).

g) Preparation of 2-[4-(4-trifluoromethylbenzyl)-1-piperazinyl]-2,4,6-cycloheptyl from α'-bromo-α,α,α-trifluoroxylene Trien- ₁ - _one ; mp 111-113°C; HRMS ( _FAB ): MH ⁺ calcd for _C19H19F3N3O 349.1529, found 349.1528.

h) Preparation of 2-[4-(4-nitrobenzyl)-1-piperazinyl]-2,4,6-cycloheptatrien-1-one from 4-nitrobenzyl bromide; mp108-110 ° ^{C; HRMS (FAB): MH+} _{calcd for C18H19N3O3 326.1506 ,} found _326.1512 .

i) Preparation of 2-[4-(3-cyanobenzyl)-1-piperazinyl]-2,4,6-cycloheptatrien-1-one from α-bromo-m-toluonitrile.

j) Preparation of 2-[4-(3-trifluoromethylbenzyl)-1-piperazinyl]-2,4,6 from α'-bromo-α,α,α-trifluoro-m-xylene - Cycloheptatrien-1-one; MS (FAB) 349 (M ⁺ 1).

k) Preparation of 2-[4-(4-bromobenzyl)-1-piperazinyl]-2,4,6-cycloheptatrien-1-one from 4-bromobenzyl bromide; mp122-124°C; MS (FAB) 359 (M ⁺ +1).

l) Preparation of 2-(2-methyl-4-naphthyl-1-piperazinyl)-2,4,6-cycloheptatriene-1-one from 2-(bromomethyl)-naphthalene; MS( FAB) 331 (M ⁺⁺ 1).

m) Preparation of 2-[4-(3,5-difluorobenzyl)-1-piperazinyl]-2,4,6-cycloheptatriene-1 from α-bromo-3,5-difluorotoluene -ketone.

n) Preparation of 2-[4-(2-cyanobenzyl)-1-piperazinyl]-2,4,6-cycloheptatrien-1-one from α-bromo-o-toluonitrile.

o) Preparation of 2-[4-(2-chlorobenzyl)-1-piperazinyl]-2,4,6-cycloheptatrien-1-one from 2-chlorobenzyl bromide.

p) Preparation of 2-[4-(2-nitrobenzyl)-1-piperazinyl]-2,4,6-cycloheptatrien-1-one from 2-nitrobenzyl bromide.

q) Preparation of 2-[4-(3-methylbenzyl)-1-piperazinyl]-2,4,6-cycloheptatrien-1-one from α-bromo-m-xylene.

r) Preparation of 2-{4-[1-(6,7-dimethoxycoumarin)methyl]-1- from 4-(bromomethyl)-6,7-dimethoxycoumarin Piperazinyl}-2,4,6-cycloheptatrien-1-one.

s) Preparation of 2-{4-[1-(7-methoxycoumarin)methyl]-1-piperazinyl}-2 from 4-(bromomethyl)-7-methoxycoumarin ,4,6-Cycloheptatrien-1-one.

t) Preparation of 2-[4-(2-phenylbenzyl)-1-piperazinyl]-2,4,6-cycloheptatrien-1-one from 2-(bromomethyl)biphenyl.

u) Preparation of 2-[4-(3-bromobenzyl)-1-piperazinyl]-2,4,6-cycloheptatrien-1-one from 3-bromobenzyl bromide.

v) Preparation of 2-{4-[3,5-bis(trifluoromethyl)benzyl]-1-piperazinyl}-2,4 from 3,5-bis(trifluoromethyl)benzyl bromide, 6-Cycloheptatrien-1-one.

w) Preparation of 2-[4-(3-nitrobenzyl)-1-piperazinyl]-2,4,6-cycloheptatrien-1-one from 3-nitrobenzyl bromide.

x) Preparation of 2-[4-(4-chlorobenzyl)-1-piperazinyl]-2,4,6-cycloheptatrien-1-one from 4-chlorobenzyl bromide; HRMS(FAB): MH ⁺ calcd _{for C19H18ClN2O} 315.1264, found 315.1270 _.

y) Preparation of 2-[4-(3-chlorobenzyl)-1-piperazinyl]-2,4,6-cycloheptatrien-1-one from 3-chlorobenzyl bromide; HRMS(FAB): MH ₊ calcd _{for C19H18ClN2O} 315.1264, found 315.1258 ^.

z) Preparation of 7-bromo-2-[ 4-(4-Chlorobenzyl)-1-piperazinyl]-2,4,6-cycloheptatrien-1-one.

Example 4(a) 2-(4-piperonyl-1-piperazinyl)-2,4,6-cycloheptatrien-1-one, maleate

2-(4-Piperonyl-1-piperazinyl)-2,4,6-cycloheptatrien-1-one (0.949 g, 2.92 mmol, Example 1a) was dissolved in diethyl ether (15.0 mL) and added A solution of maleic acid (0.373 g, 3.22 mmol) in methanol (0.50 mL). The resulting mixture was stirred at room temperature for 16 hours. The mixture was filtered to give the maleate salt (1.09 g, 85%) as a yellow solid; mp 142-144°C; MS (ES) 325 (M ⁺ -C ₄ H ₄ O ₄ ).

In a similar manner, the maleate salts of the following other compounds were prepared:

b) 2-[4-(4-trifluoromethoxybenzyl)-1-piperazinyl]-2,4,6-cycloheptatrien-1-one (Example 3f); mp170-172°C ; HRMS (FAB): MH ⁺ calcd for _{C19H19F3N2O2 (} free base _{) 365.1523} , found 365.1477 _.

c) 2-(2-Methyl-4-naphthyl-1-piperazinyl)-2,4,6-cycloheptatrien-1-one (Example 31); m.p. 170-172°C.

Example 5 2-(4-piperonyl-1-piperazinyl)-7-thiophen-2-yl-2,4,6-cycloheptatrien-1-one

To a solution of 2-bromothiophene (1.0 g, 6.1 mmol) in tetrahydrofuran (50 mL) was added n-butyllithium (1.6M in cyclohexane, 4.0 mL) at -78°C and the resulting solution was stirred for 5 minutes. At this point, _ZnCl2 (0.5M in THF, 2.0 mL) was added and the solution was allowed to warm to room temperature over 2 hours. Tetrakis(triphenylphosphine)palladium (200 mg) and 7-bromo-2-(4-piperonyl-piperazinyl)-2,4,6-cycloheptatrien-1-one ( 300 mg, 0.74 mmol, of the acetate salt of Example 1b) in THF (5 mL) and the resulting mixture was refluxed for 18 hours. The reaction was quenched with saturated aqueous ammonium chloride and the product was extracted with dichloromethane (3 x 200 mL). The crude product was purified by silica gel chromatography to give the title compound (50 mg, 50%) as a dark yellow oil.

Example 6 Receptor Binding Assay

The compounds of Examples 1 and 2 were evaluated for their binding affinity to D2 and D4 receptors based on their reduced ability to bind to ³ H-spirophenazone compared to the reference compound clozapine. The potency of a test compound to reduce binding of ³ H-heliperidine is directly related to its affinity for binding to the receptor.

D4 receptor preparation

HEK 298 (human embryonic kidney) cells stably transfected with human D4 receptor (D4.2 subtype) were cultured for 5 days (75% confluent) in a NUNC cell culture incubator and removed with versene ( approximately 19 mg cells per cell culture plate). The pellet was then centrifuged at 5000 rpm (GS3 rotor) for 10 minutes in a Sorval centrifuge and the pellet was snap frozen in liquid nitrogen and stored at -80°C until use in the binding assay. When used in the assay, cells were thawed on ice for 20 minutes before addition of 10 ml incubation buffer (50 mM Tris, 1 mM EDTA, 4 mM _MgCl2 , 5 mM KCl, 1.5 mM _CaCl2 , 120 mM NaCl, pH 7.4). Cells were then vortexed to resuspend the pellet and homogenized with a Kinematica CH-6010 Kriens-LU homogenizer at setting 7 for 15 seconds. The concentration of receptor protein was determined by Pierce BCA assay.

D2 receptor preparations

_GH4C1 (rat pituitary gland) cells stably transfected with human D2 receptor (short isoform) _were cultured in HAM'S F10 medium in a NUNC cell culture incubator for 5 days. Add 100 μM ZnSO ₄ (D2 promoter is zinc inducible) to the cells. After 16 hours, fresh medium was added to allow the cells to recover for 24 hours. Cells were harvested with versine and centrifuged at 5000 rpm (GS3 rotor) for 10 minutes in a Sorval centrifuge. Pellets were snap frozen in liquid nitrogen and stored at -80 °C until used in binding assays. When used in the assay, cells were thawed on ice for 20 minutes. Each cell culture incubator produced approximately 72 mg protein. 10 ml incubation buffer was added to the pellet, which was vortexed, resuspended and homogenized with a Kinematica CH-6010 Kriens-LU homogenizer at setting 7 for 15 seconds. The concentration of receptor protein was determined by Pierce BCA assay.

Total spirophenazone binding assay

By adding 100 μl (50 μg protein) membrane homogenate to 300 μl incubation buffer and 100 μl (0.25 nM final concentration) ³ H-heliperidine (90 Ci/mmol Amersham (in Diluted in borosilicate glass vials)) solution to start the incubation. The plate was vortexed and incubated at room temperature for 90 minutes. The binding reaction was stopped by filtration with a Packard Harvester. Samples were vacuum filtered on glass fiber filter plates (whatmanGF/B) presoaked for 2 hours with 0.3% polyethyleneimine (PEI) in 50 mM Tris buffer, pH 7.4. The filtrate was then washed 6 times with 7 ml of ice-cold 50 mM Tris buffer (pH 7.4). Filter plates were dried overnight and 35 [mu]l Microscint-O (Packard) added. The plates were sealed and counted with a Packard Top Count (3 minutes per well).

Non-specific binding assay for D4

By adding 100 μl (50 μg protein) membrane homogenate to 200 μl incubation buffer in a 96-well polypropylene plate (1 mL per well), 100 μl ³ H-helicoperidine (90 Ci/mmol Amersham (in borosilicate glass vials) Diluted to a final concentration of 0.25 nM)) and 100 μl (final concentration 30 μM) of fresh dopamine (ResearchBiochemical Inc., photoprotected and dissolved in incubation buffer) solution to start the incubation. The plate was vortexed and incubated at room temperature for 90 minutes at which time the binding reaction was stopped by filtration. The filtrate was washed and non-specific binding (NSB) was counted in the same way as described in the total binding assay above.

Non-specific binding assay of D2

The assay used the same method as in the non-specific binding assay of D4 except that 2 μM (final concentration) (-) sulpiride (Research Chemicals Inc.) was used instead of dopamine.

surrogate binding assay

By adding 100 μl (50 μg protein) membrane homogenate to 200 μl incubation buffer in a 96-well polypropylene plate (1 mL per well), 100 μl (final concentration 0.25 nM) ³ H-heliperidine (90 Ci/mmol Amersham (in Borosilicate glass vials)) and 100 μl of test compound solutions prepared from 1 mM stock solutions dissolved in DMSO were initiated and stored in polypropylene cryogenic storage vials at -20°C until used in 96-well polypropylene plates. Dilute in incubation buffer. The plate was vortexed and incubated at room temperature for 90 minutes at which time the binding reaction was stopped by filtration. Filtrates were washed and counted to obtain surrogate binding values (B _D ) in the same manner as described above in the total binding assay.

calculate

Test compounds were tested initially at 1 and 0.1 [mu]M and then over a range of concentrations selected so that intermediate doses would cause 50% inhibition ^of3H -spirulina binding. Specific binding (B _O ) in the absence of test compound is the difference between total binding (B _T ) minus nonspecific binding (NBS), and similarly specific binding (in the presence of test compound) (B) is the surrogate Difference in binding (B _D ) minus non-specific binding (NBS). _IC50 was determined from inhibition response curves of log-log plots of %B/ _B0 vs. concentration of test compound.

Calculation of Ki by Cheng and Prustoff conversion formula

Ki=IC ₅₀ /(1+[L]/K _D ) where [L] is the concentration of ³ H-helicone used in the assay and K _D is ^{the 3} H-helix measured separately under the same binding conditions Dissociation constant of phenidine.

The results of the assay are given in the table below, which clearly show the superior D4 selectivity or binding affinity of the compounds of the present invention over clozapine.

实施例7功能测定D4 affinity and selectivity

Embodiment 7 functional measurement

D4 receptors respond to dopamine and other antagonists by reducing adenylate cyclase-mediated production of cyclic AMP. The ability of a particular test compound to reverse dopamine inhibition of adenylyl cyclase was determined by the following method. Forskolin was used to increase basal adenylate cyclase activity.

CHO Pro 5 cells stably expressing human D4.2 receptors were plated in DMEM (Dulbecco's modified Eagle medium)/F12 (nutrient mixture F12 (Ham )) medium in a 6-well plate and cultured at 37°C in a CO ₂ incubator. Cells were grown to approximately 70% confluency prior to use in the assay.

Antagonist assay

The medium in each well was removed, and the wells were washed once with serum-free medium (SFM) (DMEM/F12 medium). Add 2ml of fresh SFM+IBMX medium (SFM containing 0.5mM IBMX, 3-isobutyl-1-methylxanthine, 0.1% ascorbic acid and 10μM parjiline) to each well, and then incubate at 37°C and CO ₂ Incubate for 10 minutes in the incubator. After culturing, remove the SFM+IBMX medium and add fresh SFM+IBMX medium to the wells containing one of the following: a) forskolin (final concentration 10 μM); b) dopamine and forskolin (final concentration and c) test compounds (10 ⁻⁴ to 10 ⁻⁶ ), and dopamine and forskolin (both final concentrations 10 μM). Basal adenylyl cyclase activity was determined from wells to which only SFM+IBMX medium was added.

Cells were incubated for 30 min at 37 °C in a CO ₂ incubator. After culturing, the medium was removed from each well, followed by washing once with 1 mL of PBS (phosphate-buffered saline). It was then treated with 1 mL of cold 95% ethanol:5 mM EDTA (2:1) for 1 hour at 4°C. Cells were then scraped from each well and transferred to individual Eppendorf tubes. Centrifuge the tube for 5 min at 4 °C and transfer the supernatant to a new Eppendorf tube. The pellet was discarded and the supernatant was stored at 4°C until determination of cAMP concentration. The cAMP content in fmol/well of each extract was determined by EIA (enzyme-immunoassay) using the Amersham Biotrak cAMPEIA kit (Amersham RPN 225).

The total inhibition (I ₀ ) of dopamine on forskolin-stimulated adenylyl cyclase activity was determined as the concentration of cAMP in forskolin-treated cells (C _f ) compared to the concentration of dopamine-forskolin-treated cells ( C _d ) difference.

I ₀ =C _f -C _d

The net inhibition of forskolin-stimulated adenylyl cyclase activity by dopamine in the absence of antagonists was determined (I) as the concentration of cAMP in forskolin-treated cells (C _f ) compared with the test compound, dopamine and forskolin Difference in concentration (C) of laryngin-treated cells.

I=C _f -C

The ability of a test compound to reverse dopamine suppression (% Reversal, %R) is determined by the following formula:

%R=(1-I/I ₀ )X100 compound % reversal of dopamine effect 1 μM 10 μM Clozapine 6 58 2-(4-Piperonyl-1-piperazinyl)-2,4,6-cycloheptatrien-1-one 32 65 2-(4-Hexyl-1-piperazinyl)-2,4,6-cycloheptatrien-1-one 45 50

Agonist assay

To CHO cells stably expressing D4.2 prepared as described above, test compounds and forskolin (final concentration 10 μM) were added. Cell culture, extraction and determination of cAMP concentration were performed as above. Agonist activity of the test compound will result in a decrease in cAMP concentration compared to cells treated with forskolin alone ( _Cf ). However, no reduction was observed, so the test compound did not exhibit dopamine agonist activity. Other compounds of the invention are also predicted to exhibit dopamine antagonist activity based on structural and biological functional similarities.

Claims (22)

1. formula I compound, its steric isomer, its solvate, or its pharmacologically acceptable salt:

R wherein ₁Be H, halogen, aryl or independently be selected from halogen, C by one or two _1-4Alkyl, C _1-4Alkoxyl group, nitro, trifluoromethyl, the aryl that the group of trifluoromethoxy or cyano group replaces;

R ₂Be C _4-9Alkyl; Phenyl or independently be selected from OH, cyano group, halogen, C by one or two _1-4Alkoxyl group, C _1-4Alkyl, trifluoromethyl, trifluoromethoxy, the phenyl that nitro or phenyl groups replace; Naphthyl; With 5 or 6 element heterocycle condensed phenyl, tonka bean camphor part

R wherein ₃And R ₄Independently be selected from H, C _1-4Alkyl and C _1-4Alkoxyl group; Or 1, the 2-methylenedioxyphenyl;

Condition is to work as R ₁When being H, R ₂Not phenyl or 3, the 4-Dimethoxyphenyl.

2. according to the compound of claim 1, R wherein ₂Be selected from methyl by one or two, methoxyl group, nitro, bromine, chlorine, fluorine, trifluoromethoxy, the phenyl that the group of trifluoromethyl or cyano group replaces; Phenyl; Naphthyl; Or with 5 or 6-element heterocycle condensed phenyl.

3. according to the compound of claim 1, R wherein ₂It is the 4-aminomethyl phenyl; The 4-Trifluoromethoxyphen-l; The 4-trifluoromethyl; The 4-nitrophenyl; The 3-cyano-phenyl; The 3-trifluoromethyl; The 4-bromophenyl; 3, the 5-difluorophenyl; The 2-cyano-phenyl; The 2-chloro-phenyl-; The 2-nitrophenyl; The 3-aminomethyl phenyl; The 2-phenyl; The 3-bromophenyl; 3,5-two (trifluoromethyl) phenyl; The 3-nitrophenyl; The 4-cyano-phenyl; The 3-p-methoxy-phenyl; The 2-naphthyl; The 3-chloro-phenyl-; 4-chloro-phenyl-or 1, the 2-methylenedioxyphenyl.

4. according to the compound of claim 3, R wherein ₁Be H, halogen or aryl.

5. according to the compound of claim 4, R wherein ₁Be H, bromine or thiophene.

6. according to the compound of claim 5, R wherein ₁Be connected the 7-position of 7-person's ring.

7. according to the compound of claim 6, R wherein ₁Be thiophene or R ₂Be 1, the 2-methylenedioxyphenyl.

8. according to the compound of claim 1, R wherein ₂Be the tonka bean camphor part, and R ₄And R ₅Be independently selected from H or C _1-4Alkoxyl group.

9. compound according to Claim 8, wherein R ₂Be 1-(6, the 7-escoparone) or 1-(ayapanin).

10. according to the compound of claim 9, R wherein ₁Be H.

11. according to the compound of claim 1, wherein compound is

2-(4-piperonyl-1-piperazinyl)-2,4,6-cycloheptatriene-1-ketone;

2-(4-hexyl-1-piperazinyl)-2,4,6-cycloheptatriene-1-ketone;

2-[4-(3-methoxy-benzyl)-1-piperazinyl]-2,4,6-cycloheptatriene-1-ketone;

2-[4-(4-cyano group benzyl)-1-piperazinyl]-2,4,6-cycloheptatriene-1-ketone;

2-[4-(4-methyl-benzyl)-1-piperazinyl]-2,4,6-cycloheptatriene-1-ketone;

2-[4-(4-trifluoro-methoxybenzyl)-1-piperazinyl]-2,4,6-cycloheptatriene-1-ketone;

2-[4-(4-trifluoromethyl benzyl)-1-piperazinyl]-2,4,6-cycloheptatriene-1-ketone;

2-[4-(4-nitrobenzyl)-1-piperazinyl]-2,4,6-cycloheptatriene-1-ketone;

2-[4-(3-cyano group benzyl)-1-piperazinyl]-2,4,6-cycloheptatriene-1-ketone;

2-[4-(3-trifluoromethyl benzyl)-1-piperazinyl]-2,4,6-cycloheptatriene-1-ketone;

2-[4-(4-bromobenzyl)-1-piperazinyl]-2,4,6-cycloheptatriene-1-ketone;

2-[4-(4-naphthyl-2-methyl)-1-piperazinyl]-2,4,6-cycloheptatriene-1-ketone;

2-[4-(3, the 5-difluorobenzyl)-1-piperazinyl]-2,4,6-cycloheptatriene-1-ketone;

2-[4-(2-benzyl chloride base)-1-piperazinyl]-2,4,6-cycloheptatriene-1-ketone;

2-[4-(3-methyl-benzyl)-1-piperazinyl]-2,4,6-cycloheptatriene-1-ketone;

2-[4-(2-phenylbenzyl)-1-piperazinyl]-2,4,6-cycloheptatriene-1-ketone;

2-[4-(3-bromobenzyl)-1-piperazinyl]-2,4,6-cycloheptatriene-1-ketone;

2-{4-[3,5-two (trifluoromethyl) benzyl]-the 1-piperazinyl }-2,4,6-cycloheptatriene-1-ketone;

2-[4-(3-nitrobenzyl)-1-piperazinyl]-2,4,6-cycloheptatriene-1-ketone;

2-[4-(3-benzyl chloride base)-1-piperazinyl]-2,4,6-cycloheptatriene-1-ketone;

2-[4-(4-benzyl chloride base)-1-piperazinyl]-2,4,6-cycloheptatriene-1-ketone;

7-bromo-2-[4-(4-benzyl chloride base)-1-piperazinyl]-2,4,6-cycloheptatriene-1-ketone;

7-bromo-2-(4-piperonyl-1-piperazinyl)-2,4,6-cycloheptatriene-1-ketone;

2-(4-piperonyl-1-piperazinyl)-7-thiophene-2-base-2,4,6-cycloheptatriene-1-ketone;

2-{4-[1-(6, the 7-escoparone) methyl]-the 1-piperazinyl }-2,4,6-cycloheptatriene-1-ketone; Or

2-{4-[1-(ayapanin) methyl]-the 1-piperazinyl }-2,4,6-cycloheptatriene-1-ketone.

12. according to the compound of claim 11, wherein compound is

2-(4-piperonyl-1-piperazinyl)-2,4,6-cycloheptatriene-1-ketone;

2-(4-hexyl-1-piperazinyl)-2,4,6-cycloheptatriene-1-ketone;

2-[4-(3-methoxy-benzyl)-1-piperazinyl]-2,4,6-cycloheptatriene-1-ketone;

2-[4-(4-cyano group benzyl)-1-piperazinyl]-2,4,6-cycloheptatriene-1-ketone;

2-[4-(4-methyl-benzyl)-1-piperazinyl]-2,4,6-cycloheptatriene-1-ketone;

2-[4-(4-trifluoro-methoxybenzyl)-1-piperazinyl]-2,4,6-cycloheptatriene-1-ketone;

2-[4-(4-trifluoromethyl benzyl)-1-piperazinyl]-2,4,6-cycloheptatriene-1-ketone;

2-[4-(4-nitrobenzyl)-1-piperazinyl]-2,4,6-cycloheptatriene-1-ketone;

2-[4-(3-trifluoromethyl benzyl)-1-piperazinyl]-2,4,6-cycloheptatriene-1-ketone;

2-[4-(4-bromobenzyl)-1-piperazinyl]-2,4,6-cycloheptatriene-1-ketone;

2-[4-(4-naphthyl-2-methyl)-1-piperazinyl]-2,4,6-cycloheptatriene-1-ketone;

2-[4-(3-bromobenzyl)-1-piperazinyl]-2,4,6-cycloheptatriene-1-ketone;

2-[4-(3-benzyl chloride base)-1-piperazinyl]-2,4,6-cycloheptatriene-1-ketone;

2-[4-(4-benzyl chloride base)-1-piperazinyl]-2,4,6-cycloheptatriene-1-ketone;

7-bromo-2-(4-piperonyl-1-piperazinyl)-2,4,6-cycloheptatriene-1-ketone; Or

2-(4-piperonyl-1-piperazinyl)-7-thiophene-2-base-2,4,6-cycloheptatriene-1-ketone.

13. according to the compound of claim 12, wherein compound is

2-(4-piperonyl-1-piperazinyl)-2,4,6-cycloheptatriene-1-ketone;

2-[4-(3-methoxy-benzyl)-1-piperazinyl]-2,4,6-cycloheptatriene-1-ketone;

2-[4-(4-cyano group benzyl)-1-piperazinyl]-2,4,6-cycloheptatriene-1-ketone;

2-[4-(4-bromobenzyl)-1-piperazinyl]-2,4,6-cycloheptatriene-1-ketone;

2-[4-(4-trifluoro-methoxybenzyl)-1-piperazinyl]-2,4,6-cycloheptatriene-1-ketone;

2-[4-(4-trifluoromethyl benzyl)-1-piperazinyl]-2,4,6-cycloheptatriene-1-ketone;

2-[4-(4-nitrobenzyl)-1-piperazinyl]-2,4,6-cycloheptatriene-1-ketone;

2-[4-(3-benzyl chloride base)-1-piperazinyl]-2,4,6-cycloheptatriene-1-ketone;

2-[4-(4-benzyl chloride base)-1-piperazinyl]-2,4,6-cycloheptatriene-1-ketone;

2-[4-(4-methyl-benzyl)-1-piperazinyl]-2,4,6-cycloheptatriene-1-ketone;

7-bromo-2-(4-piperonyl-1-piperazinyl)-2,4,6-cycloheptatriene-1-ketone; Or

2-[4-(4-naphthyl-2-methyl)-1-piperazinyl]-2,4,6-cycloheptatriene-1-ketone.

14. according to the compound of claim 13, wherein compound is

2-(4-piperonyl-1-piperazinyl)-2,4,6-cycloheptatriene-1-ketone;

2-[4-(4-bromobenzyl)-1-piperazinyl]-2,4,6-cycloheptatriene-1-ketone;

2-[4-(4-trifluoromethyl benzyl)-1-piperazinyl]-2,4,6-cycloheptatriene-1-ketone;

2-[4-(4-trifluoro-methoxybenzyl)-1-piperazinyl]-2,4,6-cycloheptatriene-1-ketone;

2-[4-(4-nitrobenzyl)-1-piperazinyl]-2,4,6-cycloheptatriene-1-ketone;

2-[4-(4-benzyl chloride base)-1-piperazinyl]-2,4,6-cycloheptatriene-1-ketone;

7-bromo-2-(4-piperonyl-1-piperazinyl)-2,4,6-cycloheptatriene-1-ketone; Or

2-[4-(4-naphthyl-2-methyl)-1-piperazinyl]-2,4,6-cycloheptatriene-1-ketone.

15. according to the compound of claim 1, wherein R ₁Be H and R ₂By a halogen, cyano group, alkoxyl group, trifluoromethoxy, trifluoromethyl, or C _1-4The phenyl that alkyl substituent replaces; Or 1, the 2-methylenedioxyphenyl.

16. a pharmaceutical composition comprises pharmaceutically acceptable carrier and the formula II compound for the treatment of significant quantity:

R wherein ₁Be independently selected from H, halogen, aryl or independently be selected from halogen, C by one or two _1-4Alkyl, C _1-4Alkoxyl group, nitro, trifluoromethyl, the aryl that the group of trifluoromethoxy or cyano group replaces;

R ₅Be independently selected from C _4-9Alkyl; Phenyl or independently be selected from OH, cyano group, halogen, C by one or two _1-4Alkoxyl group, C _1-4Alkyl, trifluoromethyl, trifluoromethoxy, the phenyl that nitro or phenyl groups replace; Naphthyl; With 5 or 6 element heterocycle condensed phenyl; With the tonka bean camphor part

R wherein ₃And R ₄Independently be selected from H, C _1-4Alkyl and C _1-4Alkoxyl group.

17. the pharmaceutical composition of claim 16, condition are to work as R ₁When being H, R ₂Not phenyl or 3, the 4-Dimethoxyphenyl.

18. claim 1 compound, the optional dopamine D that is used to prepare the antagonism patient that combines with pharmaceutical carrier ₄The purposes of the pharmaceutical composition of receptor acting.

19. claim 1 compound, the optional combination with pharmaceutical carrier is used to prepare the purposes for the treatment of schizoid pharmaceutical composition.

20. claim 1 compound, the optional purposes that is used to prepare the pharmaceutical composition for the treatment of anxiety that combines with pharmaceutical carrier.

21. claim 1 compound is as pharmaceutical active compounds.

22. preparation, its steric isomer, solvate, or the method for pharmacologically acceptable salt:

R wherein ₁Be H, halogen, aryl or independently be selected from halogen, C by one or two _1-4Alkyl, C _1-4Alkoxyl group, nitro, trifluoromethyl, the aryl that the group of trifluoromethoxy or cyano group replaces;

R ₂Be C _4-9Alkyl; Phenyl or independently be selected from OH, cyano group, halogen, C by one or two _1-4Alkoxyl group, C _1-4Alkyl, trifluoromethyl, trifluoromethoxy, the phenyl that nitro or phenyl groups replace; Naphthyl; With 5 or 6 element heterocycle condensed phenyl, tonka bean camphor part

R wherein ₃And R ₄Independently be selected from H, C _1-4Alkyl and C _1-4Alkoxyl group; Or 1, the 2-methylenedioxyphenyl;

Condition is to work as R ₁When being H, R ₂Not phenyl or 3, the 4-Dimethoxyphenyl;

This method may further comprise the steps:

The reagent of formula A

Reagent with formula B

Coupling in backflow methyl alcohol obtains formula I compound.