CN106146404B - Pyridazinone derivative and application thereof - Google Patents

Abstract

The present invention relates to the field of medicine, in particular, the present invention relates to pyridazinone derivatives and applications thereof, more particularly, the present invention relates to pyridazinone derivatives, pharmaceutical compositions comprising the pyridazinone derivatives and the pyridazinone derivatives The composition and the use of the pyridazinone derivative in the preparation of a medicament for preventing or treating pain diseases. Pyridazinone derivatives have the structure of formula (I). It has been found through experiments that the compounds can be used to prevent or treat pain-related diseases.

Description

Pyridazinone derivatives and their applications

technical field

The present invention relates to the field of medicine, in particular, the present invention relates to pyridazinone derivatives and applications thereof, more particularly, the present invention relates to pyridazinone derivatives, pharmaceutical compositions comprising the pyridazinone derivatives and the pyridazinone derivatives The composition and the use of the pyridazinone derivative in the preparation of a medicament for preventing or treating pain diseases.

Background technique

Pain is a basic sense that humans are born with, and it is of great significance in protecting the human body from injury and maintaining the internal environment of the human body. However, long-term or excessive pain can seriously affect people's normal physiology and life. Chronic pain and neuralgia have been defined in medicine as diseases that seriously endanger human health. Among them, neuralgia is one of the pain diseases that seriously endanger the lives of patients. About 8% of the people in the world are affected by this disease. Although researchers have done a lot of related work in the fields of basic research and clinical research, there are still huge challenges in clinical research on the pathogenesis of neuralgia and its analgesic mechanism. There are no broad-spectrum and specific targeted drugs for neuralgia. The drugs used clinically for the treatment of neuralgia are mainly drugs with neuralgia analgesic activity for the treatment of depression, epilepsy and other diseases.

The sigma-1 receptor (σ ₁ receptor) is an emerging drug target in recent years, especially showing excellent potential in neuralgia analgesia. σ ₁ receptors are mainly distributed in the central nervous system, but also widely distributed in peripheral organs. Research on the regulation of sigma ₁ receptors in human pain began in the 1990s, when it was found that sigma ₁ receptor antagonists can sensitize the analgesic activity of opioid analgesics, while sigma ₁ agonists attenuate this phenomenon. In acute pain experiments, antisense oligodeoxynucleotides of σ ₁ receptors can significantly enhance the analgesic effects of morphine and opioid μ receptor agonists. More experiments have demonstrated that σ ₁ receptors act through direct interaction with opioid μ receptors. On the other hand, it has been demonstrated that σ ₁ receptor antagonists themselves also have analgesic effect on pain, especially neuralgia. At present, the forefront of sigma-1 receptor antagonist research is S1RA, which has high affinity for sigma- ₁ receptors and high selectivity for sigma- ₁ receptors. At present, S1RA is used in experiments for the treatment of various pains and has entered the clinical stage, among which the experiment for the treatment of neuralgia alone has entered the clinical phase II.

Therefore, the search for σ ₁ receptor antagonists with selectivity to σ ₁ receptors for anti-pain therapy has important scientific value and social significance for clinical treatment of pain and neuralgia.

SUMMARY OF THE INVENTION

The present invention aims to solve at least one of the technical problems existing in the prior art. Therefore, an object of the present invention is to propose a sigma ₁ receptor antagonist selective for sigma ₁ receptor which can be used for anti-pain therapy.

According to one aspect of the present invention, the present invention provides a compound, which is a compound represented by formula I or a stereoisomer, tautomer, nitrogen oxide, solvate, metabolite, A pharmaceutically acceptable salt or its prodrug

in,

R ₁ is optionally substituted C _1-5 alkyl, optionally substituted C _3-7 cycloalkyl, optionally substituted aryl, optionally substituted thienyl, optionally substituted indolyl, the said The substituted substituent is selected from at least one of alkyl, cyano, hydroxyl, halogen, -CN, -N(CN) ₂ and -C(CN) ₃ ;

Z is optionally substituted -R _c -R _d -, R _c is O, S, NH or CH ₂ , R _d is a straight-chain or branched-chain hydrocarbon group or a heterohydrocarbyl group with 1 to 10 carbon atoms, wherein any Preferably, the straight-chain or branched-chain hydrocarbon group or hetero-hydrocarbon group with 1 to 10 carbon atoms contains at least one oxygen atom or ethylene group, and the substituted substituent is selected from alkyl, cyano, hydroxyl, halogen, at least one of -CN, -N(CN) ₂ and -C(CN) ₃ ;

Q is N or CH, R _a and R _b are each independently an optionally substituted straight-chain or branched alkyl group containing 1 to 5 carbon atoms, and the substituted substituent is selected from alkyl, cyano, hydroxyl , at least one of halogen, -CN, -N(CN) ₂ and -C(CN) ₃ ,

Alternatively Q, together with the Ra and Rb attached thereto, together form a five- to seven-membered ring containing at least one O, N or carbonyl group, the five- to seven-membered ring being optionally substituted with an optionally substituted alkane containing 1 to 5 carbon atoms base, optionally substituted aryl, optionally substituted thiophene, C _1-10 alkyl formate substituted, and the substituted substituent is selected from alkyl, cyano, hydroxyl, halogen, -CN, -N At least one of (CN) ₂ and -C(CN) ₃ .

The inventors have surprisingly found that the compounds of the present invention have selective antagonism to σ ₁ receptors and have the potential for analgesic activity, ie have the potential to treat pain, especially neuralgia. Further, through animal experiments, the inventors found that the compounds of the present invention can be effectively used in the treatment and prevention of various pain diseases including acute pain, chronic pain, intractable pain, cancer pain and special pain.

According to another aspect of the present invention, the present invention also provides a pharmaceutical composition. According to an embodiment of the present invention, the pharmaceutical composition contains the aforementioned compound. The inventors have surprisingly found that the pharmaceutical composition of the present invention containing the aforementioned compounds with selective antagonism to σ ₁ receptors can be effectively used in the treatment and prevention of acute pain, chronic pain, intractable pain, cancer pain and special Pain and other pain-related diseases.

According to another aspect of the present invention, the present invention also provides the use of the aforementioned compound or pharmaceutical composition in the preparation of a medicament for preventing or treating pain-related diseases.

Detailed ways

Embodiments of the present invention are described in detail below. The embodiments described below are exemplary, only for explaining the present invention, and should not be construed as limiting the present invention.

Definitions and General Terms

Unless otherwise stated, the following definitions as used herein shall apply. For the purposes of the present invention, chemical elements are in accordance with the CAS version of the Periodic Table of the Elements, and the Handbook of Chemistry and Physics (75th Edition, 1994). In addition, general principles of organic chemistry may be referred to as described in "Organic Chemistry", Thomas Sorrell, University Science Books, Sausalito: 1999, and "March's Advanced Organic Chemistry" by Michael B. Smith and Jerry March, John Wiley & Sons, New York: 2007, Its entire contents are incorporated herein by reference.

The term "patient" as used herein refers to humans (including adults and children) or other animals. According to some embodiments of the invention, "patient" refers to a human.

The term "stereoisomers" refers to compounds that have the same chemical structure, but differ in the arrangement of atoms or groups in space. Stereoisomers include enantiomers, diastereomers, conformational isomers (rotamers), geometric isomers (cis/trans), atropisomers, etc. .

The term "enantiomer" refers to two nonsuperimposable, but mirror-image isomers of a compound.

The term "diastereomer" refers to a stereoisomer having two or more chiral centers and whose molecules are not mirror images of each other. Diastereomers have different physical properties such as melting point, boiling point, spectral properties and reactivity. Diastereomeric mixtures can be separated by high resolution analytical procedures such as electrophoresis and chromatography, eg HPLC.

The term "chirality" refers to a molecule that has the property of being non-superimposable with its mirror image; whereas "achiral" refers to a molecule that is superimposable with its mirror image.

The term "racemate" or "racemic mixture" refers to an equimolar mixture of two enantiomers lacking optical activity.

Stereochemical definitions and rules used in the present invention generally follow S.P. Parker, Ed., McGraw-Hill Dictionary of Chemical Terms (1984) McGraw-Hill Book Company, New York; and Eliel, E. and Wilen, S, "Stereochemistry of Organic Compounds" ", John Wiley & Sons, Inc, New York, 1994. Many organic compounds exist in optically active forms, that is, they have the ability to rotate the plane of polarized light. In describing optically active compounds, the prefixes D and L or R and S are used to denote the absolute configuration of the molecule about one or more of its chiral centers. The prefixes d and 1 or (+) and (-) are symbols used to designate the rotation of plane polarized light by the compound, where (-) or 1 indicates that the compound is levorotatory. Compounds prefixed with (+) or d are dextrorotatory. A specific stereoisomer is an enantiomer, and a mixture of such isomers is called an enantiomeric mixture. A 50:50 mixture of enantiomers is called a racemic mixture or racemate, which can occur when there is no stereoselectivity or stereospecificity in a chemical reaction or process.

Any asymmetric atom (eg, carbon, etc.) of the compounds disclosed herein may exist in racemic or enantiomerically enriched forms, such as (R)-, (S)- or (R,S)-configurations exist. In certain embodiments, each asymmetric atom has at least 50% enantiomeric excess, at least 60% enantiomeric excess, at least 70% enantiomeric excess, at least 70% enantiomeric excess in the (R)- or (S)-configuration 80% enantiomeric excess, at least 90% enantiomeric excess, at least 95% enantiomeric excess, or at least 99% enantiomeric excess.

Depending on the choice of starting materials and process, the compounds of the present invention may be present as one of the possible isomers or as mixtures thereof, such as racemates and mixtures of diastereomers (depending on the number of asymmetric carbon atoms) form exists. Optically active (R)- or (S)-isomers can be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques. If the compound contains a double bond, the substituent may be in the E or Z configuration; if the compound contains a disubstituted cycloalkyl, the cycloalkyl substituent may have the cis or trans configuration.

The resulting mixtures of any stereoisomers may be separated into pure or substantially pure geometric isomers, enantiomers, diastereomers on the basis of differences in the physicochemical properties of the components, for example, by chromatography and/or fractional crystallization.

Any resulting racemate of the final product or intermediate can be resolved into the optical enantiomers by known methods by methods familiar to those skilled in the art, eg, by performing diastereomeric salts thereof obtained. separation. Racemic products can also be separated by chiral chromatography, eg, high performance liquid chromatography (HPLC) using chiral adsorbents. In particular, enantiomers can be prepared by asymmetric synthesis, for example, see Jacques, et al., Enantiomers, Racemates and Resolutions (Wiley Interscience, New York, 1981); Principles of Asymmetric Synthesis (2 ^nd Ed. Robert E. Gawley, Jeffrey Aube, Elsevier, Oxford, UK, 2012); Eliel, EL Stereochemistry of Carbon Compounds (McGraw-Hill, NY, 1962); Wilen, SH Tables of Resolving Agents and Optical Resolutions p.268 (ELEliel, Ed., Univ. of NotreDame Press, Notre Dame, IN 1972); Chiral Separation Techniques: A Practical Approach (Subramanian, G. Ed., Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim, Germany, 2007).

The term "tautomer" or "tautomeric form" refers to structural isomers of different energies that are interconvertible through a low energy barrier. A chemical equilibrium of tautomers can be achieved if tautomerism is possible (eg, in solution). For example, protontautomers (also known as prototropic tautomers) include interconversions by migration of protons, such as keto-enol isomerization and imine-ene Amine isomerization. Valence tautomers include interconversions by recombination of some of the bonding electrons. A specific example of keto-enol tautomerism is the interconversion of pentane-2,4-dione and 4-hydroxypent-3-en-2-one tautomers. Another example of tautomerism is phenol-ketone tautomerism. A specific example of phenol-ketone tautomerism is the interconversion of pyridin-4-ol and pyridin-4(lH)-one tautomers. Unless otherwise indicated, all tautomeric forms of the compounds of the present invention are within the scope of the present invention.

The term "optional" or "optionally" means that the subsequently described event or circumstance may but need not occur, and that the description includes instances where said event or circumstance occurs and instances where it does not. For example, "optional bond" means that the bond may or may not be present, and that the description includes single, double, or triple bonds.

The term "comprising" is an open-ended expression, that is, it includes the contents specified in the present invention, but does not exclude other aspects.

The term "unsaturated" or "unsaturated" means that a moiety contains one or more degrees of unsaturation.

As described herein, the compounds of the present invention may be optionally substituted with one or more substituents, such as the compounds of the general formula above, or as in the Examples, subclasses, and subclasses encompassed by the present invention. a class of compounds. It is to be understood that the term "optionally substituted" is used interchangeably with the term "substituted or unsubstituted". In general, the term "substituted" means that one or more hydrogen atoms in a given structure have been replaced with a specified substituent. Unless otherwise indicated, an optional substituent group may be substituted at each substitutable position of the group. When more than one position in a given formula can be substituted with one or more substituents selected from a particular group, the substituents can be substituted identically or differently at each position.

In addition, it should be noted that, unless it is clearly stated otherwise, the description mode "...respectively and independently" used in the present invention should be understood in a broad sense, which may refer to different groups and between the same symbols. The specific options expressed do not affect each other, and it can also be expressed that in the same group, the specific options expressed between the same symbols do not affect each other.

In various parts of this specification, the substituents of the compounds disclosed in the present invention are disclosed in terms of group type or scope. Specifically, the present invention includes each and every independent subcombination of each member of these group species and ranges. For example, the term " _C1-5 alkyl" specifically refers to the independently disclosed methyl, ethyl, _C3 alkyl, _C4 alkyl and _C5 alkyl groups.

Unless explicitly described, the term "alkyl" or "alkyl group" as used in the present invention refers to a saturated linear or branched monovalent hydrocarbon group containing 1 to 20 carbon atoms, wherein the alkyl group A group can be optionally substituted with one or more substituents described herein. Unless otherwise specified, alkyl groups contain 1-20 carbon atoms. According to one embodiment of the present invention, the alkyl group contains 1-12 carbon atoms; according to another embodiment of the present invention, the alkyl group contains 1-6 carbon atoms; according to one embodiment of the present invention, the alkane group The radical group contains 1-4 carbon atoms; according to another embodiment of the present invention, the alkyl group contains 1-3 carbon atoms. Examples of alkyl groups include, but are not limited to, methyl (Me, _-CH3 ), ethyl (Et, -CH2CH3), n _{- propyl} (n _- Pr, _{-CH2CH2CH3 )} ), isopropyl (i-Pr, -CH(CH ₃ ) ₂ ), n-butyl (n-Bu, -CH ₂ CH ₂ CH ₂ CH ₃ ), isobutyl (i-Bu, -CH ₂ CH ) (CH ₃ ) ₂ ), sec-butyl (s-Bu, -CH(CH ₃ )CH ₂ CH ₃ ), tert-butyl (t-Bu, -C(CH ₃ ) ₃ ), n-pentyl (-CH 2CH2CH2CH2CH3), ₂ -pentyl (-CH( _{CH3 ) CH2CH2CH3 )} , _{3 -} pentyl (-CH( _{CH2CH3 ) 2 )} , ₂ -methyl -2-butyl(-C(CH3)2CH2CH3), ₃ -methyl- _{2-butyl(-CH(CH3)CH(CH3)2 ) , 3 - methyl} -1- Butyl ( _-CH2CH2CH ( _CH3 ) ₂ ), ₂ -methyl- ₁ -butyl (-CH2CH( _CH3 ) _{CH2CH3 )} , n _- hexyl _{( -CH2CH2CH2 CH2CH2CH3} ), ₂ -hexyl (-CH( _CH3 ) _{CH2CH2CH2CH3 )} , _{3 -} hexyl (-CH( _{CH2CH3 ) ( CH2CH2CH3 ) )} , 2-methyl-2-pentyl(-C( _CH3 )2CH2CH2CH3), ₃ -methyl- ₂ -pentyl(-CH( _{CH3 ) CH} ( _{CH3 ) CH2CH3} ), 4-methyl-2-pentyl (-CH(CH ₃ )CH ₂ CH(CH ₃ ) ₂ ), 3-methyl-3-pentyl (-C(CH ₃ )(CH ₂ CH ₃ ) ₂ ), 2-methyl-3-pentyl (-CH(CH ₂ CH ₃ )CH(CH ₃ ) ₂ ), 2,3-dimethyl-2-butyl (-C(CH ₃ ) ₂ CH (CH ₃ ) ₂ ), 3,3-dimethyl-2-butyl (-CH(CH ₃ )C(CH ₃ ) ₃ ), n-heptyl, n-octyl, and the like.

The term "carbonyl", whether used alone or in combination with other terms, such as "aminocarbonyl" or "acyloxy", means -(C=O)-.

The term "H" refers to a single hydrogen atom. Such radicals may be attached to other groups, such as oxygen atoms, to form hydroxyl groups.

The term "halogen" refers to fluorine (F), chlorine (Cl), bromine (Br) or iodine (I).

The term "alkoxy" means that an alkyl group is attached to the remainder of the molecule through an oxygen atom, wherein the alkyl group has the meaning as described herein. Unless otherwise specified, the alkoxy groups contain 1-12 carbon atoms. According to an embodiment of the present invention, the alkoxy group contains 1-6 carbon atoms; according to an embodiment of the present invention, the alkoxy group contains 1-4 carbon atoms; according to an embodiment of the present invention, Alkoxy groups contain 1-3 carbon atoms. The alkoxy group is optionally substituted with one or more substituents described herein.

Examples of alkoxy groups include, but are not limited to, methoxy (MeO, -OCH ₃ ), ethoxy (EtO, -OCH ₂ CH ₃ ), 1-propoxy (n-PrO, n- Propoxy, -OCH ₂ CH ₂ CH ₃ ), 2-propoxy (i-PrO, i-propoxy, -OCH(CH ₃ ) ₂ ), 1-butoxy (n-BuO, n- Butoxy, -OCH ₂ CH ₂ CH ₂ CH ₃ ), 2-methyl-l-propoxy (i-BuO, i-butoxy, -OCH ₂ CH(CH ₃ ) ₂ ), 2-butanyl Oxy (s-BuO, s-butoxy, -OCH(CH ₃ )CH ₂ CH ₃ ), 2-methyl-2-propoxy (t-BuO, t-butoxy, -OC(CH ₃ ) ₃ ), ₁ -pentyloxy (n _- pentyloxy, -OCH2CH2CH2CH2CH3), _{2 -} pentyloxy (-OCH _{( CH3 ) CH2CH2CH3 )} , 3-pentyloxy (-OCH(CH ₂ CH ₃ ) ₂ ), 2-methyl-2-butoxy (-OC(CH ₃ ) ₂ CH ₂ CH ₃ ), 3-methyl-2-butoxy group (-OCH(CH ₃ )CH(CH ₃ ) ₂ ), 3-methyl-1-butoxy (-OCH ₂ CH ₂ CH(CH ₃ ) ₂ ), 2-methyl-1-butoxy (-OCH ₂ CH(CH ₃ )CH ₂ CH ₃ ), etc.

The term "ring" includes carbocyclic, heterocyclic, aromatic, heteroaromatic, and the like, wherein the carbocyclic, heterocyclic, aromatic, and heteroaromatic groups have the meaning as described herein.

The term "cycloalkyl" denotes a monovalent or polyvalent saturated monocyclic, bicyclic or tricyclic ring system containing 3 to 12 carbon atoms. Bicyclic or tricyclic ring systems may include fused, bridged, and spiro rings. According to one embodiment of the present invention, the cycloalkyl group contains 3-10 carbon atoms; according to one embodiment of the present invention, the cycloalkyl group contains 3-8 carbon atoms; according to one embodiment of the present invention, the cycloalkyl group contains 3-6 carbon atoms. Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like. The cycloalkyl group is optionally substituted with one or more substituents described herein.

The term "aryl" refers to monocyclic, bicyclic and tricyclic carbocyclic ring systems containing 6-14 ring atoms, or 6-12 ring atoms, or 6-10 ring atoms, at least one of which is aromatic of. The aryl group is usually, but not necessarily, attached to the parent molecule through the aromatic ring of the aryl group. Examples of aryl groups may include phenyl, naphthyl, and anthracene. The aryl group is optionally substituted with one or more substituents described herein.

The term "prodrug" as used in the present invention refers to the conversion of a compound into a compound of formula (I) in vivo. Such conversion is effected by hydrolysis of the prodrug in blood or enzymatic conversion to the parent structure in blood or tissue. A detailed discussion of prodrugs can be found in the following references: Higuchi et al., Pro-drugs as Novel Delivery Systems, Vol. 14, A.C.S. Symposium Series; Roche et al., ed., Bioreversible Carriersin Drug Design, American Pharmaceutical Association and Pergamon Press, 1987; Rautio et al., Prodrugs: Design and Clinical Applications, Nature Reviews Drug Discovery, 2008, 7, 255-270, and Hecker et al, Prodrugs of Phosphates and Phosphonates, J. Med. Chem., 2008, 51, 2328-2345 , each document is hereby incorporated by reference.

The term "metabolite" as used in the present invention refers to a product obtained by metabolism of a specific compound or its salt in vivo. Metabolites of a compound can be identified by techniques well known in the art, and their activity can be characterized using assays as described herein. Such products may be obtained by subjecting the administered compound to oxidation, reduction, hydrolysis, amidation, deamidation, esterification, delipidation, enzymatic cleavage, and the like. Accordingly, the present invention includes metabolites of compounds, including metabolites produced by contacting a compound of the present invention with a mammal for a sufficient period of time.

As used herein, "pharmaceutically acceptable salts" refer to organic and inorganic salts of the compounds of the present invention. Pharmaceutically acceptable salts are well known in the art as described in: SM Berge et al., J. Pharmaceutical Sciences, 66: 1-19, 1977. Pharmaceutically acceptable non-toxic acid salts include, but are not limited to, inorganic acid salts formed by reaction with amino groups including hydrochloride, hydrobromide, phosphate, sulfate, perchlorate, And organic acid salts such as acetate, oxalate, maleate, tartrate, citrate, succinate, malonate, or obtained by other methods such as ion exchange method described in books and literature these salts. Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, besylate, benzoate, bisulfate, borate, butyrate, camphoric acid Salt, camphorsulfonate, cyclopentylpropionate, digluconate, lauryl sulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate Salt, Gluconate, Hemisulfate, Heptanoate, Caproate, Hydroiodide, 2-Hydroxy-ethanesulfonate, Lacturonate, Lactate, Laurate, Lauryl Sulfate, Malonate, Malonate, Mesylate, 2-Naphthalenesulfonate, Nicotinate, Nitrate, Oleate, Palmitate, Pamoate, Pectate, Persulfate, 3 - Phenylpropionate, picrate, pivalate, propionate, stearate, thiocyanate, p-toluenesulfonate, undecanoate, valerate, etc. Salts obtained with appropriate bases include alkali metal, alkaline earth metal, ammonium and N ⁺ (C _1-4 alkyl) ₄ salts. The present invention also contemplates quaternary ammonium salts formed from any compound containing an N-group. Water- or oil-soluble or dispersible products can be obtained by quaternization. Alkali metal or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like. Pharmaceutically acceptable salts further include appropriate, non-toxic ammonium, quaternary ammonium salts and amine cations that resist the formation of counterions, such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, _{C1 -8} Sulfonates and aromatic sulfonates.

A "solvate" of the present invention refers to an association of one or more solvent molecules with a compound of the present invention. Solvate-forming solvents include, but are not limited to, water, isopropanol, ethanol, methanol, dimethyl sulfoxide, ethyl acetate, acetic acid, aminoethanol. The term "hydrate" refers to an association in which the solvent molecule is water.

When the solvent is water, the term "hydrate" may be used. According to one embodiment of the present invention, one molecule of the compound of the present invention may be combined with one molecule of water, such as a monohydrate; according to an embodiment of the present invention, one molecule of the compound of the present invention may be combined with more than one molecule of water, Like a dihydrate, according to one embodiment of the present invention, one molecule of a compound of the present invention may be associated with less than one molecule of water, such as a hemihydrate. It should be noted that the hydrates of the present invention retain the bioavailability of the non-hydrated form of the compounds.

The term "treating" any disease or disorder, as used herein, in some embodiments refers to ameliorating the disease or disorder (ie, slowing or arresting or alleviating the development of the disease or at least one clinical symptom thereof). In other embodiments, "treating" refers to alleviating or improving at least one bodily parameter, including bodily parameters that may not be perceived by a patient. In other embodiments, "treating" refers to modulating a disease or disorder physically (eg, stabilizing an observable symptom) or physiologically (eg, stabilizing a physical parameter), or both. In other embodiments, "treating" refers to preventing or delaying the onset, occurrence or worsening of a disease or disorder.

The term "prevention" refers to the reduction of the risk of acquiring a disease or disorder (ie: stopping the development of at least one clinical symptom of the disease in a subject who may be facing or predisposed to facing the disease, but has not experienced or manifested it yet) symptoms of the disease).

The pharmaceutically acceptable salts of the present invention can be synthesized from the parent compound, basic or acidic moieties, using conventional chemical methods. In general, such salts can be prepared by reacting the free acid forms of these compounds with a stoichiometric amount of a suitable base (eg, hydroxide, carbonate, bicarbonate, etc. of Na, Ca, Mg, or K), or by The preparations are carried out by reacting the free base forms of these compounds with a stoichiometric amount of the appropriate acid. Such reactions are usually carried out in water or an organic solvent or a mixture of the two. Generally, where appropriate, the use of non-aqueous media such as diethyl ether, ethyl acetate, ethanol, isopropanol or acetonitrile is required. In, for example, "Remington's Pharmaceutical Sciences", 20th Edition, Mack Publishing Company, Easton, Pa., (1985); and "Handbook of Pharmaceutical Salts: Properties, Selection, and Use )", Stahl and Wermuth (Wiley-VCH, Weinheim, Germany, 2002) can find additional lists of suitable salts.

In addition, the compounds disclosed herein, including their salts, can also be obtained in their hydrate form or in a form containing a solvent (eg, ethanol, DMSO, etc.) for their crystallization. Compounds of the present disclosure may form solvates, inherently or by design, with pharmaceutically acceptable solvents, including water; thus, the present invention is intended to include both solvated and unsolvated forms.

Any structural formula given herein is also intended to represent both isotopically unenriched as well as isotopically enriched forms of these compounds. Isotopically enriched compounds have the structures depicted by the general formulae given herein, except that one or more atoms are replaced by atoms having a selected atomic weight or mass number. Exemplary isotopes that can be incorporated into the compounds of the present invention include isotopes of ^hydrogen , carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, and chlorine, such as 2H, ^3H , ^11C , ^13C , ^14C , ^15N , ^17O , ¹⁸ O, ¹⁸ F, ³¹ P, ³² P, ³⁵ S, ³⁶ Cl and ¹²⁵ I.

In another aspect, the compounds of the present invention include isotopically enriched compounds as defined by the present invention, for example, those in which radioactive isotopes, such as ³ H, ¹⁴ C and ¹⁸ F are present, or in which non-radioactive isotopes, such as ² H and ¹³ C. Such isotopically enriched compounds can be used in metabolic studies (using ¹⁴ C), reaction kinetic studies (using eg ² H or ³ H), detection or imaging techniques such as positron emission tomography (PET) or including drugs or Single photon emission computed tomography (SPECT) for substrate tissue distribution measurements, or may be used in radiation therapy of patients. ¹⁸ F-enriched compounds are particularly ideal for PET or SPECT studies. Isotopically enriched compounds can be prepared by conventional techniques familiar to those skilled in the art or as described in the Examples and Preparations of the present invention, using a suitable isotopically labeled reagent in place of the unlabeled reagent originally used.

compound

According to one aspect of the present invention, the present invention provides a compound, which is a compound represented by formula I or a stereoisomer, tautomer, nitrogen oxide, solvate, metabolite, A pharmaceutically acceptable salt or its prodrug

in,

R ₁ is optionally substituted C _1-5 alkyl, optionally substituted C _3-7 cycloalkyl, optionally substituted aryl, optionally substituted thienyl, optionally substituted indolyl, the said The substituted substituent is selected from at least one of alkyl, cyano, hydroxyl, halogen, -CN, -N(CN) ₂ and -C(CN) ₃ ;

Z is optionally substituted -R _c -R _d -, R _c is O, S, NH or CH ₂ , R _d is a straight-chain or branched-chain hydrocarbon group or a heterohydrocarbyl group with 1 to 10 carbon atoms, wherein any Preferably, the straight-chain or branched-chain hydrocarbon group or hetero-hydrocarbon group with 1 to 10 carbon atoms contains at least one oxygen atom or ethylene group, and the substituted substituent is selected from alkyl, cyano, hydroxyl, halogen, at least one of -CN, -N(CN) ₂ and -C(CN) ₃ ;

Q is N or CH, R _a and R _b are each independently an optionally substituted straight-chain or branched alkyl group containing 1 to 5 carbon atoms, and the substituted substituent is selected from alkyl, cyano, hydroxyl , at least one of halogen, -CN, -N(CN) ₂ and -C(CN) ₃ ,

Alternatively Q, together with the Ra and Rb attached thereto, together form a five- to seven-membered ring containing at least one O, N or carbonyl group, the five- to seven-membered ring being optionally substituted with an optionally substituted alkane containing 1 to 5 carbon atoms base, optionally substituted aryl, optionally substituted thiophene, C _1-10 alkyl formate substituted, and the substituted substituent is selected from alkyl, cyano, hydroxyl, halogen, -CN, -N At least one of (CN) ₂ and -C(CN) ₃ .

The inventors have surprisingly found that the compounds of the present invention have selective antagonism to σ ₁ receptors and have the potential for analgesic activity, ie have the potential to treat pain, especially neuralgia. Further, through animal experiments, the inventors found that the compounds of the present invention can be effectively used in the treatment and prevention of various pain diseases including acute pain, chronic pain, intractable pain, cancer pain and special pain.

Wherein, it should be noted that the expression "R ₁ is optionally substituted C _1-5 alkyl, optionally substituted C _3-7 cycloalkyl, optionally substituted aryl, optionally substituted aryl, Substituted thienyl, optionally substituted indolyl, the substituted substituent is selected from at least one of alkyl, cyano, hydroxyl, halogen, -CN, -N(CN) ₂ and -C(CN) ₃ In "one", the term "substituted substituent" refers to when C _1-5 alkyl, C _3-7 cycloalkyl, aryl, thienyl, and indolyl are substituted, used to replace C _1-5 Substituents for alkyl, substituents for C _3-7 cycloalkyl, substituents for aryl, substituents for thienyl, and substituents for indolyl are each independently At least one selected from the group consisting of alkyl, cyano, hydroxyl, halogen, -CN, -N(CN) ₂ and -C(CN) ₃ .

The expression "Z is optionally substituted -R _c -R _d -, R _c is O, S, NH or CH ₂ , and R _d is a straight or branched chain having 1 to 10 carbon atoms. Hydrocarbyl or heterohydrocarbyl, wherein optionally the straight or branched chain hydrocarbyl or heterohydrocarbyl having 1 to 10 carbon atoms contains at least one oxygen atom or ethylene group, and the substituted substituent is selected from alkyl groups , cyano, hydroxyl, halogen, -CN, -N(CN) ₂ and -C(CN) ₃ " at least one", the term "substituted substituent" refers to when -R _c -R _d - is replaced by When substituted, the substituent for -R _c -R _d - is selected from at least one of alkyl, cyano, hydroxyl, halogen, -CN, -N(CN) ₂ and -C(CN) ₃ .

The expression "Q is N or CH, R _a and R _b are each independently an optionally substituted linear or branched alkyl group containing 1 to 5 carbon atoms, and the substituted substituents are selected from From at least one of alkyl, cyano, hydroxyl, halogen, -CN, -N(CN) ₂ and -C(CN) ₃ ", the term "substituted substituent" refers to when it contains 1 to 5 When the straight-chain or branched-chain alkyl group of carbon atoms is substituted, the substituent used to replace the straight-chain or branched-chain alkyl group containing 1 to 5 carbon atoms is selected from alkyl, cyano, hydroxyl, halogen, -CN, At least one of -N(CN) ₂ and -C(CN) ₃ .

As used herein, the expression "or Q, together with Ra and Rb attached thereto, together form a five- to seven-membered ring containing at least one O, N or carbonyl group, the five- to seven-membered ring being optionally substituted with an optionally substituted containing 1-5 carbon atoms alkyl, optionally substituted aryl, optionally substituted thiophene, C _1-10 alkyl formate substituted, and the substituted substituent is selected from alkyl, cyano, hydroxyl , halogen, at least one of -CN, -N(CN) ₂ and -C(CN) ₃ ", the term "substituted substituent" refers to an alkyl group, an aryl group containing 1 to 5 carbon atoms when , thiophene, and C _1-10 alkyl formate groups are substituted, the substituents used to replace alkyl groups containing 1 to 5 carbon atoms, the substituents used to replace aryl groups, and the substituents used to replace thiophene , The substituents used to replace the C _1-10 alkyl formate group are independently selected from alkyl, cyano, hydroxyl, halogen, -CN, -N(CN) ₂ and -C(CN) ₃ at least one.

Regarding "alkyl, cyano, hydroxyl, halogen, -CN, -N(CN) ₂ and -C(CN) ₃ " unless explicitly stated, the definitions are as commonly understood by those skilled in the art and as defined in the present invention of.

According to an embodiment of the present invention, R ₁ is an optionally substituted cyclopentyl group, an optionally substituted phenyl group, an optionally substituted naphthyl group, an optionally substituted thienyl group, an optionally substituted indolyl group, and the The substituted substituent is selected from at least one of alkyl, cyano, hydroxyl, halogen, -CN, -N(CN) ₂ and -C(CN) ₃ . The term "substituted substituent" as used herein refers to the substituents used to replace cyclopentyl, the substituents used to replace benzene when cyclopentyl, phenyl, naphthyl, thienyl, and indolyl are substituted The substituents for substituted naphthyl, the substituents for substituted naphthyl, the substituents for thienyl, and the substituents for indolyl are independently selected from alkyl, cyano, hydroxyl, halogen, -CN, At least one of -N(CN) ₂ and -C(CN) ₃ .

According to an embodiment of the present invention, Z is an optionally substituted alkoxy group containing 1 to 5 carbon atoms, and the substituted substituent is selected from alkyl, cyano, hydroxyl, halogen, -CN, -N(CN ) ₂ and at least one of -C(CN) ₃ . The term "substituted substituent" as used herein means that when the alkoxy group containing 1 to 5 carbon atoms is substituted, the substituent for replacing the alkoxy group containing 1 to 5 carbon atoms is selected from alkyl groups , at least one of cyano, hydroxyl, halogen, -CN, -N(CN) ₂ and -C(CN) ₃ .

According to an embodiment of the present invention, Q together with its attached _Ra and _Rb forms

或者

or

wherein R ₂ and R ₃ are each independently optionally substituted C _1-5 alkyl, R ₄ and R ₅ are each independently selected from hydrogen, optionally substituted C _1-5 alkyl, hydroxy, tert-butoxy One or more of carbonyl and carbonyl, wherein the substituted substituent is selected from at least one of alkyl, cyano, hydroxyl, halogen, -CN, -N(CN) ₂ and -C(CN) ₃ one;

m is 0, 1 or 2; and

X is one of oxygen, nitrogen, or CH.

The substituted substituent is at least one selected from alkyl, cyano, hydroxyl, halogen, -CN, -N(CN) ₂ and -C(CN) ₃ .

It should be noted that, with respect to R ₂ and R ₃ , the term "substituted substituent" used herein means that when the C _1-5 alkyl group is substituted, the substituent for replacing the C _1-5 alkyl group is selected from the group consisting of _At least one of alkyl, cyano, hydroxyl, halogen, -CN, -N(CN) ₂ and -C(CN) _{3 ;} for R4 and R5, when _R4 and _R5 are optionally substituted In the case of a C _1-5 alkyl group, the term "substituted substituent" as used herein means that when the C _1-5 alkyl group is substituted, the substituent for replacing the C _1-5 alkyl group is selected from the group consisting of alkyl, cyano at least one of radical, hydroxyl, halogen, -CN, -N(CN) ₂ and -C(CN) ₃ ; for R ₅ , when R ₅ is optionally substituted C _1-5 alkyl, hydroxyl, tertiary When one or more of butoxycarbonyl and carbonyl are used, the term "substituted substituent" used herein means that when C _1-5 alkyl, hydroxyl, tert-butoxycarbonyl and carbonyl are substituted, the Substituents for C _1-5 alkyl, substituents for hydroxyl, substituents for tert-butoxycarbonyl, substituents for carbonyl are independently selected from alkyl, cyano, hydroxyl, halogen At least one of , -CN, -N(CN) ₂ and -C(CN) ₃ .

According to an embodiment of the present invention, the halogen is fluorine, chlorine, bromine and iodine.

According to an embodiment of the present invention, R ₁ is phenyl, dichlorophenyl, benzyl, fluorophenyl, chlorophenyl, naphthyl or cyclopentyl;

Z is ethoxy, propoxy or butoxy;

Q is N, CH, Ra and Rb are each independently methyl, ethyl, propyl, isopropyl, or

Q together with its attached Ra and Rb forms a five- to seven-membered ring containing at least one O, N or carbonyl group, optionally with methyl, propyl, phenyl, tert-butyl formate replace.

According to an embodiment of the present invention, the compound of the present invention is a compound represented by the general formula (I) or a pharmaceutically acceptable salt thereof:

in,

Z is substituted or unsubstituted -O(CH ₂ ) _n -, n is an integer from 2 to 4, and the substituted substituent hydroxy or methyl, or the carbon chain in Z contains a double bond or an oxygen atom;

R ₁ is hydrogen, substituted or unsubstituted C _1-5 alkyl, substituted or unsubstituted C _3-7 cycloalkyl, substituted or unsubstituted aryl, wherein the substituted substituent is selected from alkyl, One or more of cyano, hydroxyl or halogen;

Q is N or CH, R _a and R _b are each independently an optionally substituted straight-chain or branched alkyl group containing 1 to 5 carbon atoms, and the substituted substituent is selected from alkyl, cyano, hydroxyl , at least one of halogen, -CN, -N(CN) ₂ and -C(CN) ₃ ,

Alternatively Q, together with the Ra and Rb attached thereto, together form a five- to seven-membered ring containing at least one O, N or carbonyl group, the five- to seven-membered ring being optionally substituted with an optionally substituted alkane containing 1 to 5 carbon atoms base, optionally substituted aryl, optionally substituted thiophene, C _1-10 alkyl formate substituted, and the substituted substituent is selected from alkyl, cyano, hydroxyl, halogen, -CN, -N At least one of (CN) ₂ and -C(CN) ₃ .

According to an embodiment of the present invention, in the compound of general formula (I), the unsubstituted C _1-5 alkyl group is selected from methyl, ethyl, propyl, isopropyl, butyl, isobutyl, pentyl or isopentyl, the substituted C _1-5 alkyl group is selected from halogen substituted C _1-5 alkyl groups, the halogen being fluorine, chlorine, bromine, iodine.

According to an embodiment of the present invention, in the compound of general formula (I), the unsubstituted C _3-7 cycloalkyl group is selected from cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, substituted C _{3-- 7} cycloalkyl is selected from halogen-substituted C _3--7 cycloalkyl, C _1-5 alkyl substituted C _3--7 cycloalkyl, and the halogen is fluorine, chlorine, bromine, iodine; the C _1-5 alkyl is one or more of methyl, ethyl, propyl, isopropyl, butyl, isobutyl, pentyl or isopentyl.

According to an embodiment of the present invention, in the compound of general formula (I), the substituted or unsubstituted aryl group is selected from substituted or unsubstituted phenyl group, substituted or unsubstituted benzyl group, substituted or unsubstituted naphthyl group , the substituent is selected from one or more of alkyl, cyano, hydroxyl or halogen.

According to an embodiment of the present invention, in the compound of general formula (I), the halogen is fluorine, chlorine, bromine and iodine.

According to an embodiment of the present invention, in the compound of general formula (I), the substituted phenyl group is methylphenyl, methoxyphenyl, fluorophenyl, and chlorophenyl; the substituted benzyl group is methoxy benzyl.

According to an embodiment of the present invention, the compound is at least one of the following compounds, or a stereoisomer, tautomer, nitrogen oxide, solvate, metabolite, pharmacy of at least one of the following compounds On an acceptable salt or its prodrug:

According to some embodiments of the present invention, the present invention also includes the compound of formula (I) and the salts of the above specific compounds, the salts are salts containing pharmaceutically acceptable anions: such as hydrochloride, hydrobromide, Hydroiodide, Nitrate, Sulfate or Bisulfate, Phosphate or Acid Phosphate, Acetate, Lactate, Citrate, Tartrate, Maleate, Fumarate, Methanesulfonate acid salt, gluconate, saccharate, benzoate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, etc.

use

According to another aspect of the present invention, the present invention also provides a pharmaceutical composition. According to an embodiment of the present invention, the pharmaceutical composition contains the aforementioned compound. The inventors have surprisingly found that the pharmaceutical composition of the present invention containing the aforementioned compounds with selective antagonism to σ ₁ receptors can be effectively used in the treatment and prevention of acute pain, chronic pain, intractable pain, cancer pain and special Pain and other pain-related diseases.

According to an embodiment of the present invention, the pharmaceutical composition of the present invention further comprises a pharmaceutically acceptable excipient, carrier, adjuvant, vehicle or a combination thereof.

According to an embodiment of the present invention, the pharmaceutical composition of the present invention further comprises a drug for preventing or treating pain-related diseases different from the aforementioned compounds, and said drugs for preventing or treating pain-related diseases different from the aforementioned compounds It is at least one selected from the following: non-steroidal anti-inflammatory painkillers, central painkillers, narcotic painkillers, and traditional Chinese medicine compound painkillers.

According to other embodiments of the present invention, the drugs for preventing or treating pain diseases different from the aforementioned compounds are at least one selected from the group consisting of aspirin, ibuprofen, indomethacin, paracetamol, phenylbutazone , rofecoxib, celecoxib, tramadol, fentanyl, morphine, domeridine, anisodamine, naproxen, fenbid, voltaren and analgesic new.

According to another aspect of the present invention, the present invention also provides the use of the aforementioned compound or pharmaceutical composition in the preparation of a medicament for preventing or treating pain-related diseases.

According to an embodiment of the present invention, the pain-like disease is neuralgia.

According to an embodiment of the present invention, the pharmaceutical composition of the present invention comprises a compound of formula (I) or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable adjuvant (such as a carrier and/or excipient, etc.), the A pharmaceutical composition is a compound containing sufficient pain, neuralgia analgesic activity.

According to embodiments of the present invention, an effective dose of a compound of the present invention may be administered orally together with, for example, an inert diluent or some kind of carrier. According to some embodiments of the present invention, the compounds of the present invention may be encapsulated in gelatin capsules or compressed into tablets. For the purpose of oral therapy, the compounds of the present invention can be used with excipients and in the form of tablets, troches, capsules, suspensions, syrups and the like. According to embodiments of the present invention, the above formulations should contain at least 0.5% by weight of the active compound of the present invention, but may vary depending on the particular dosage form, where from 4% to about 70% by weight per unit is convenient. The amount of active compound in such pharmaceutical compositions should result in an appropriate dosage. An oral unit dose of preferred pharmaceutical compositions and formulations of the present invention contains 1.0-300 mg of the active compound of the present invention.

According to the embodiments of the present invention, the compounds provided by the present invention and their pharmaceutically acceptable salts, solvates and hydrates can be combined with pharmaceutically acceptable carriers or diluents to form pharmaceutical preparations. Suitable pharmaceutically acceptable carriers include inert solid fillers or diluents and sterile aqueous or organic solutions.

According to embodiments of the present invention, the amount of the compound of the present invention will depend on the type and severity of the disease or condition, and also on the characteristics of the subject, such as general health, age, sex, weight and drug tolerance. The skilled artisan will be able to determine appropriate dosages based on these and other factors. Effective doses of commonly used CNS drugs are well known to the skilled artisan. The total daily dose is usually between about 0.05 mg and 2000 mg.

According to embodiments of the present invention, the present invention relates to pharmaceutical compositions that provide about 0.01 to 1000 mg of active ingredient per unit dose. The compositions can be administered by any suitable route, such as orally in capsule form, parenterally in the form of injection solutions, topically in the form of ointments or lotions, rectally in the form of suppositories, in the form of patch delivery systems Transdermal administration.

According to embodiments of the present invention, the compounds provided by the present invention can be combined with suitable solid or liquid carriers or diluents to form capsules, tablets, pills, powders, syrups, solutions and the like. Tablets, pills, capsules, etc. contain from about 0.01 to about 99 percent by weight of the active ingredient and binders such as gelatin, cornstarch, acacia; excipients such as calcium hydrogen phosphate; disintegrants such as cornstarch, potato starch or algae acids; lubricants such as magnesium stearate; and sweeteners such as sucrose, lactose. When the preparation is in the form of a capsule, it may contain, in addition to the above-mentioned types of materials, a liquid carrier such as a fat and oil.

According to embodiments of the present invention, when used for parenteral administration, the compounds provided herein can be combined with sterile aqueous or organic vehicles to form injectable solutions or suspensions.

The in vitro receptor binding test shows that the compounds involved in the present invention have high affinity to σ ₁ receptors, but low affinity to σ ₂ receptors. Selective antagonism of σ ₁ receptors, indicating potential for analgesic activity.

In addition, animal test results also show that these compounds can significantly improve both formalin-induced phase I and phase II pain. Since these in vitro action targets and in vivo pharmacological models are closely related to the sigma ₁ receptor-mediated nervous system regulation response, especially pain, the compounds involved in the present invention have the potential to treat pain, especially neuralgia.

Furthermore, according to other embodiments of the present invention, the compounds provided by the present invention and the pharmaceutical compositions consisting of the compounds can be used for the treatment and prevention of pain; the pain refers to acute pain, such as pain from acute injury to soft tissues and joints, pain after surgery , Obstetric pain, acute herpes zoster pain, gout, etc; , intervertebral disc herniation, intractable headache, etc.; cancer pain such as advanced tumor pain, tumor metastasis pain, etc.; special pain categories such as thromboangiitis, intractable angina, idiopathic thoracic and abdominal pain.

General Synthesis Scheme

The general method for synthesizing the compounds of the present invention is to synthesize the pyridazinone precursor first, and then connect it to the nitrogen-containing structure through a carbon chain. E.g:

The values of R1, R2, R3, R, and n are as defined above.

Synthesis Example

The following synthesis examples 1-47 are all carried out based on the above-mentioned general method, wherein the compounds prepared in each example are shown in Table 1. Example 1. Preparation of 6-(2-morpholineethoxy)-2-phenylpyridazin-3(2H)-one (compound 1)

6-(2-morpholineethoxy)-2-phenylpyridazin-3(2H)-one is prepared through the synthetic scheme shown in the above reaction formula, and the specific steps are as follows:

1) Dissolve 14.5 g of phenylhydrazine hydrochloride and 9.8 g of maleic anhydride in 200 ml of purified water. Then, 40 ml of concentrated hydrochloric acid was slowly added to the solution under stirring to obtain a reaction system. The obtained reaction system was heated to reflux and allowed to react for 6 hours. After the completion of the reaction, the reaction mixture was cooled in an ice-water bath to precipitate a yellow solid. The cooled reaction mixture was then suction filtered, and the filter cake was washed twice with water. Next, take out the washed filter cake and dissolve it with saturated NaHCO ₃ , then filter off the insolubles, and adjust the clear liquid to pH 2-3 with concentrated hydrochloric acid, so that a white solid is precipitated, and after suction filtration and drying, the obtained 17.3 g product, 92.0% yield.

2) Take 9.4 g of the product of the first step, 13.8 g of anhydrous potassium carbonate, 18.8 g of 1,2-dibromoethane, and add 100 ml of acetone to obtain a reaction system. The resulting reaction system was heated under reflux for 4 hours, and then cooled to room temperature. Next, the reaction mixture was filtered, and the solvent was evaporated to dryness to give a pale yellow oil. Then, the pale yellow oil was subjected to flash chromatography to obtain 8.5 g of a white solid with a melting point of 77-79° C. and a yield of 61.3%.

3) Take 1.40 g of the second-step product, 0.8 g of morpholine, and 2 g of potassium carbonate, and add 50 ml of acetonitrile to obtain a reaction system. The resulting reaction system was heated to reflux for 6 hours, and then cooled to room temperature. Next, the reaction mixture was evaporated to dryness of the solvent, and an appropriate amount of dichloromethane was added, followed by washing with water, and the aqueous layer was separated, the organic layer was dried over anhydrous magnesium sulfate, and the solvent was evaporated to dryness to obtain a yellow oil. Then, the pale yellow oil was subjected to flash chromatography to obtain the target compound, 1.21 g of pale yellow oil, with a yield of 80.6%.

¹ H NMR (600 MHz, CDCl ₃ ) δ 7.64 (d, J=7.7 Hz, 2H), 7.47-7.42 (m, 2H), 7.33 (t, J=7.4 Hz, 1H), 7.01 (q, J= 9.7Hz, 2H), 4.31(t, J＝5.6Hz, 2H), 3.72(t, J＝3.9Hz, 4H), 2.78(t, J＝5.6Hz, 2H), 2.56(s, 4H).MS (ESI)m/z 302.6([M+H] ⁺ )

Example 2, 6-(3-morpholinopropoxy)-2-phenylpyridazin-3(2H)-one (compound 2)

1,2-dibromoethane was replaced with 1,3-dibromopropane, and the target compound was prepared according to the method of Example 1.

¹ H NMR (600MHz, CDCl ₃ ) δ 7.65-7.63 (m, 2H), 7.45 (t, J=7.9Hz, 2H), 7.34 (t, J=7.4Hz, 1H), 7.02-6.95 (m, 2H), 4.32–4.17 (m, 2H), 3.77–3.57 (m, 4H), 2.80–2.62 (m, 1H), 2.53–2.42 (m, 5H), 2.00–1.88 (m, 2H).

MS(ESI) m/z 316.4([M+H] ⁺ )

Example 3, 6-(4-morpholinobutoxy)-2-phenylpyridazin-3(2H)-one (compound 3)

1,2-dibromoethane was replaced with 1,4-dibromobutane, and the target compound was prepared according to the method of Example 1.

¹ H NMR (600 MHz, CDCl ₃ ) δ 7.66 (d, J=7.6 Hz, 2H), 7.46 (t, J=7.9 Hz, 2H), 7.35 (t, J=7.4 Hz, 1H), 7.05-6.95 (m, 2H), 4.18 (t, J=6.4Hz, 2H), 3.72 (t, J=4.6Hz, 4H), 2.53–2.37 (m, 6H), 1.82–1.73 (m, 2H), 1.68- 1.63(m,2H).MS(ESI)m/z 330.8([M+H] ⁺ )

Example 4, 6-(3-(piperidin-1-yl)propoxy)-2-phenylpyridazin-3(2H)-one (compound 4)

Substitute 1,2-dibromoethane with 1,3-dibromopropane and morpholine with hexahydropyridine, and prepare the target compound according to the method of Example 1.

¹ H NMR (600 MHz, CDCl ₃ ) δ 7.64 (d, J=7.7 Hz, 2H), 7.44 (dd, J=10.8, 5.0 Hz, 2H), 7.35-7.30 (m, 1H), 7.00-6.94 ( m, 2H), 4.19 (t, J=6.4Hz, 2H), 2.49–2.32 (m, 6H), 2.00–1.90 (m, 2H), 1.64–1.54 (m, 4H), 1.43 (s, 2H) .MS(ESI)m/z 314.3([M+H] ⁺ )

Example 5, 6-(3-(4-methylpiperidin-1-yl)propoxy)-2-phenylpyridazin-3(2H)-one (compound 5)

Substitute 1,2-dibromoethane with 1,3-dibromopropane and morpholine with 4-methylpiperidine, and prepare the target compound according to the method of Example 1.

¹ H NMR (600 MHz, CDCl ₃ ) δ 7.66 (d, J=7.7 Hz, 2H), 7.44 (t, J=7.9 Hz, 2H), 7.33 (t, J=7.4 Hz, 1 H), 7.01-6.95 (m, 2H), 4.20 (t, J=6.4Hz, 2H), 2.90 (d, J=11.4Hz, 2H), 2.52–2.39 (m, 2H), 2.01–1.85 (m, 4H), 1.66– 1.56(m,2H),1.41-1.30(m,1H),1.24(qd,J=12.5,3.6Hz,2H),0.92(d,J=6.5Hz,3H).MS(ESI)m/z 328.4 ([M+H] ⁺ )

Example 6, 6-(3-(4-methylpiperazin-1-yl)propoxy)-2-phenylpyridazin-3(2H)-one (compound 6)

Substitute 1,2-dibromoethane with 1,3-dibromopropane and morpholine with N-methylpiperazine, and prepare the target compound according to the method of Example 1.

¹ H NMR (600 MHz, CDCl ₃ ) δ 7.62 (dd, J=8.5, 1.0 Hz, 2H), 7.43 (dd, J=11.3, 4.6 Hz, 2H), 7.35-7.31 (m, 1H), 6.98 ( q, J=9.7Hz, 2H), 4.19 (t, J=6.3Hz, 2H), 2.90–2.66 (m, 8H), 2.63–2.54 (m, 2H), 2.50 (s, 3H), 1.99–1.88 (m,2H).MS(ESI)m/z 329.4([M+H] ⁺ )

Example 7, 6-(3-(4-ethylpiperazin-1-yl)propoxy)-2-phenylpyridazin-3(2H)-one (compound 7)

Substitute 1,2-dibromoethane with 1,3-dibromopropane and morpholine with N-ethylpiperazine, and prepare the target compound according to the method of Example 1.

¹ H NMR (600 MHz, CDCl ₃ ) δ 7.61 (dd, J=8.5, 1.0 Hz, 2H), 7.41 (t, J=7.9 Hz, 2H), 7.33-7.29 (m, 1H), 7.01-6.93 ( m, 2H), 4.39–4.10 (m, 2H), 2.81–2.39 (m, 12H), 2.02–1.81 (m, 2H), 1.14 (t, J=7.3Hz, 3H).

MS(ESI) m/z 343.5([M+H] ⁺ )

Example 8, 6-(3-(3,5-dimethylpiperidin-1-yl)propoxy)-2-phenylpyridazin-3(2H)-one (compound 8)

Substitute 1,2-dibromoethane with 1,3-dibromopropane and morpholine with 3,5-dimethylpiperidine, and prepare the target compound according to the method of Example 1.

¹ H NMR (600MHz, CDCl ₃ ) δ 7.68-7.60 (m, 2H), 7.42 (t, J=7.0Hz, 2H), 7.31 (t, J=7.4Hz, 1H), 7.00-6.93 (m, 2H), 4.23–4.15 (m, 2H), 2.97–2.87 (m, 2H), 2.59–2.51 (m, 2H), 2.01 (dd, J=14.5, 7.1Hz, 2H), 1.81–1.66 (m, 2H),1.56(t,J=11.2Hz,2H),0.83(d,J=6.6Hz,6H),0.64–0.49(m,2H).MS(ESI)m/z 342.3([M+H] ⁺ )

Example 9, 6-(3-(4-methyl-1,4-diazeptan-1-yl)propoxy)-2-phenylpyridazin-3(2H)-one (compound 9 )

Substitute 1,2-dibromoethane with 1,3-dibromopropane, and replace morpholine with N-methylhomopiperazine, and prepare the target compound according to the method of Example 1.

¹ H NMR (600 MHz, CDCl ₃ ) δ 7.68-7.65 (m, 2H), 7.49 (t, J=7.4 Hz, 2H), 7.38 (t, J=7.4 Hz, 1H), 7.05 (d, J= 9.9Hz, 1H), 7.01 (d, J=9.8Hz, 1H), 4.25 (t, J=6.2Hz, 2H), 3.39–3.09 (m, 6H), 2.95 (t, J=5.9Hz, 2H) ,2.82(dd,J＝15.2,8.0Hz,2H),2.75(s,3H),2.29(s,2H),2.06–1.98(m,2H).MS(ESI)m/z 343.4([M+ H] ⁺ )

Example 10, 6-(3-(dimethylamino)propoxy)-2-phenylpyridazin-3(2H)-one (compound 10)

Substitute 1,2-dibromoethane with 1,3-dibromopropane and morpholine with dimethylamine, and prepare the target compound according to the method of Example 1.

¹ H NMR (600 MHz, CDCl ₃ ) δ 7.66-7.59 (m, 2H), 7.47-7.41 (m, 2H), 7.37-7.31 (m, 1H), 7.07-6.98 (m, 2H), 4.32-4.20 (m,2H),3.24–3.17(m,2H),2.80(s,6H),2.35–2.21(m,2H).MS(ESI)m/z 274.3([M+H] ⁺ )

Example 11, 6-(3-(diethylamino)propoxy)-2-phenylpyridazin-3(2H)-one (compound 11)

Substitute 1,2-dibromoethane with 1,3-dibromopropane and morpholine with diethylamine, and prepare the target compound according to the method of Example 1.

¹ H NMR (600 MHz, CDCl ₃ ) δ 7.67-7.63 (m, 2H), 7.49-7.44 (m, 2H), 7.35 (dd, J=10.7, 4.2 Hz, 1H), 7.04-6.98 (m, 2H) ),4.25(t,J＝6.1Hz,2H),2.96-2.84(m,6H),2.22-2.08(m,2H),1.24(t,J＝7.2Hz,6H).MS(ESI)m/ z 302.4([M+H] ⁺ )

Example 12, 6-(3-(diisopropylamino)propoxy)-2-phenylpyridazin-3(2H)-one (compound 12)

Substitute 1,2-dibromoethane with 1,3-dibromopropane and morpholine with diisopropylamine, and prepare the target compound according to the method of Example 1.

¹ H NMR (600 MHz, CDCl ₃ ) δ 7.63 (d, J=7.6 Hz, 2H), 7.37 (t, J=7.9 Hz, 2H), 7.25 (t, J=7.4 Hz, 1 H), 6.96-6.86 (m, 2H), 4.15 (t, J=6.4Hz, 2H), 2.49 (t, J=7.1Hz, 2H), 2.35–2.28 (m, 4H), 1.86–1.78 (m, 2H), 1.44– 1.32(m,4H),0.80(t,J=7.4Hz,6H).MS(ESI)m/z 330.3([M+H] ⁺ )

Example 13, 6-(3-(4-oxopiperidin-1-yl)propoxy)-2-phenylpyridazin-3(2H)-one (Compound 13)

Substitute 1,2-dibromoethane with 1,3-dibromopropane and morpholine with 4-piperidone, and prepare the target compound according to the method of Example 1.

¹ H NMR (600 MHz, CDCl ₃ ) δ 7.68-7.63 (m, 2H), 7.49-7.42 (m, 2H), 7.36-7.32 (m, J=11.9, 5.4, 1.0 Hz, 1H), 7.05-6.93 (m, 2H), 4.46–4.20 (m, 4H), 3.76–3.72 (m, 4H), 2.54–2.37 (m, 2H), 2.26–2.09 (m, 2H), 2.02–1.91 (m, 2H) .MS(ESI)m/z 328.5([M+H] ⁺ )

Example 14, 6-(4-(piperidin-1-yl)butoxy)-2-phenylpyridazin-3(2H)-one (compound 14)

Substitute 1,2-dibromoethane with 1,4-dibromobutane and morpholine with hexahydropyridine, and prepare the target compound according to the method of Example 1.

¹ H NMR (600 MHz, CDCl ₃ ) δ 7.65 (d, J=7.6 Hz, 2H), 7.48 (t, J=8.2 Hz, 2H), 7.33 (t, J=7.3 Hz, 1H), 7.05-7.02 (m, 2H), 4.25–4.21 (m, 2H), 3.15–3.06 (m, 2H), 2.97–2.88 (m, 2H), 2.78–2.70 (m, 2H), 1.98–1.53 (m, 10H) .

MS(ESI) m/z 328.5([M+H] ⁺ )

Example 15, 6-(4-(4-methylpiperazin-1-yl)butoxy)-2-phenylpyridazin-3(2H)-one (Compound 15)

Substitute 1,2-dibromoethane with 1,4-dibromobutane and morpholine with N-methylpiperazine, and prepare the target compound according to the method of Example 1.

¹ H NMR (600MHz, CDCl ₃ ) δ 7.61 (t, J=8.8Hz, 2H), 7.42 (dt, J=13.1, 5.8Hz, 2H), 7.31 (dd, J=10.1, 4.5Hz, 1H) ,7.00–6.94(m,2H),4.13(dd,J=11.6,5.4Hz,2H),2.77(t,J=50.6Hz,8H),2.61–2.42(m,5H),1.82–1.50(m ,4H).

MS(ESI) m/z 343.9([M+H] ⁺ )

Example 16, 6-(4-(4-ethylpiperazin-1-yl)butoxy)-2-phenylpyridazin-3(2H)-one (compound 16)

Substitute 1,2-dibromoethane with 1,4-dibromobutane and morpholine with N-ethylpiperazine, and prepare the target compound according to the method of Example 1.

¹ H NMR (600MHz, CDCl ₃ ) δ 7.65-7.61 (m, 2H), 7.43 (t, J=7.9Hz, 2H), 7.32 (t, J=7.4Hz, 1H), 6.95-6.90 (m, 2H), 4.43(s, 2H), 4.13(t, J=6.4Hz, 2H), 3.77(t, J=7.1Hz, 2H), 3.64-3.56(m, 2H), 2.62-2.54(m, 4H) ), 2.46–2.37 (m, 2H), 2.25 (s, 2H), 1.78–1.68 (m, 2H), 1.12 (t, J=7.2Hz, 3H).

MS(ESI) m/z 357.5([M+H] ⁺ )

Example 17, 6-(4-(diethylamino)butoxy)-2-phenylpyridazin-3(2H)-one (compound 17)

Substitute 1,2-dibromoethane with 1,4-dibromobutane and morpholine with diethylamine, and prepare the target compound according to the method of Example 1.

¹ H NMR (600 MHz, CDCl ₃ ) δ 7.68 (d, J=8.1 Hz, 2H), 7.47 (t, J=7.8 Hz, 2H), 7.37 (t, J=7.4 Hz, 1H), 7.03 (d ,J=9.7Hz,2H),6.98(d,J=9.7Hz,2H),4.25(s,2H),1.94(s,2H).MS(ESI)m/z316.8([M+H] ⁺ )

Example 18, 6-(4-(4-methyl-1,4-diazeptan-1-yl)butoxy)-2-phenylpyridazine-3(2H)-one pyridine (compound 18)

Substitute 1,2-dibromoethane with 1,4-dibromobutane, and replace morpholine with N-methylhomopiperazine, and prepare the target compound according to the method of Example 1.

¹ H NMR (600MHz, CDCl ₃ ) δ 7.84 (d, J=2.4Hz, 2H), 7.60 (d, J=2.4Hz, 2H), 7.47 (d, J=8.7Hz, 1H), 7.00-6.95 (m, 2H), 4.23 (t, J=6.1Hz, 4H), 3.51 (s, 2H), 2.85–2.74 (m, 4H), 2.52 (s, 7H), 2.18–2.05 (m, 4H). MS(ESI) m/z 357.3([M+H] ⁺ )

Example 19, 6-(4-(4-oxopiperidin-1-yl)butoxy)-2-phenylpyridazin-3(2H)-one (compound 19)

Substitute 1,2-dibromoethane with 1,4-dibromobutane and morpholine with 4-piperidone, and prepare the target compound according to the method of Example 1.

¹ H NMR (600 MHz, CDCl ₃ ) δ 7.69-7.63 (m, 2H), 7.49-7.43 (m, 2H), 7.37-7.32 (m, 1H), 7.04-6.96 (m, 2H), 4.27-4.09 (m, 2H), 3.10–2.89 (m, 2H), 2.74 (t, J=6.1Hz, 2H), 2.55–2.39 (m, 4H), 1.88–1.59 (m, 6H).

MS(ESI) m/z 342.5([M+H] ⁺ )

Example 20, 6-(4-(3,5-dimethylpiperidin-1-yl)butoxy)-2-phenylpyridazin-3(2H)-one (compound 20)

Substitute 1,2-dibromoethane with 1,4-dibromobutane and morpholine with 3,5-dimethylpiperidine, and prepare the target compound according to the method of Example 1.

¹ H NMR (600 MHz, CDCl ₃ ) δ 7.66-7.63 (m, 2H), 7.46-7.42 (m, 2H), 7.34 (t, J=7.4 Hz, 1H), 7.01-6.96 (m, 2H), 4.29–4.16 (m, 2H), 4.16–4.10 (m, 2H), 3.10 (t, J=20.3Hz, 2H), 2.74–2.64 (m, 2H), 2.07–1.86 (m, 2H), 1.85– 1.61(m,2H),0.97(d,J=8.9Hz,3H),0.83(d,J=6.6Hz,3H).MS(ESI)m/z 356.7([M+H] ⁺ )

Example 21, 2-(3,4-dichlorophenyl)-6-(3-morpholinopropoxy)pyridazin-3(2H)-one (compound 21)

Substitute phenylhydrazine hydrochloride with 3,4-dichlorophenylhydrazine hydrochloride, and prepare the target compound according to the method of Example 1.

¹ H NMR (600 MHz, CDCl ₃ ) δ 7.88 (d, J=2.4 Hz, 1H), 7.65 (dd, J=8.7, 2.5 Hz, 1H), 7.53-7.49 (m, 1H), 7.05-6.94 ( m, 2H), 4.24 (t, J=6.4Hz, 2H), 3.73 (t, J=4.5Hz, 4H), 2.51 (dd, J=16.9, 9.4Hz, 6H), 2.02–1.92 (m, 2H) ).

MS(ESI) m/z 384.8([M+H] ⁺ )

Example 22, 2-(3,4-dichlorophenyl)-6-(3-(4-methylpiperazin-1-yl)propoxy)pyridazin-3(2H)-one (Compound 22 )

Replace phenylhydrazine hydrochloride with 3,4-dichlorophenylhydrazine hydrochloride, 1,2-dibromoethane with 1,3-dibromopropane, and morpholine with N-methylpiperazine, as per example 1 to prepare the target compound.

¹ H NMR (600MHz, CDCl ₃ ) δ 7.85 (d, J=2.4Hz, 1H), 7.62 (dd, J=8.7, 2.4Hz, 1H), 7.49 (d, J=8.7Hz, 1H), 7.06 –6.93(m,2H),4.21(t,J=6.3Hz,2H),2.64(d,J=25.3Hz,8H),2.57–2.53(m,2H),2.42(s,3H),1.99– 1.90(m, 2H).

MS(ESI) m/z 397.7([M+H] ⁺ )

Example 23, 2-(3,4-dichlorophenyl)-6-(3-(piperidin-1-yl)propoxy)pyridazin-3(2H)-one (compound 23)

Replace phenylhydrazine hydrochloride with 3,4-dichlorophenylhydrazine hydrochloride, 1,2-dibromoethane with 1,3-dibromopropane, and morpholine with hexahydropyridine, as in Example 1 Preparation of the target compound.

¹ H NMR (600MHz, CDCl ₃ ) δ 7.88 (d, J=2.4Hz, 1H), 7.65 (dd, J=8.7, 2.5Hz, 1H), 7.50 (d, J=8.7Hz, 1H), 7.04 –6.96(m,2H),4.22(t,J=6.4Hz,2H),2.53-2.38(m,6H),2.02-1.92(m,2H),1.66-1.58(m,4H),1.43(d , J=30.4Hz, 2H).

MS(ESI) m/z 382.8([M+H] ⁺ )

Example 24, 2-(3,4-dichlorophenyl)-6-(3-(4-methylpiperidin-1-yl)propoxy)pyridazin-3(2H)-one (Compound 24 )

Replace phenylhydrazine hydrochloride with 3,4-dichlorophenylhydrazine hydrochloride, 1,2-dibromoethane with 1,3-dibromopropane, and morpholine with 4-methylpiperidine, according to the examples 1 to prepare the target compound.

¹ H NMR (600MHz, CDCl ₃ ) δ 7.88 (d, J=2.5Hz, 1H), 7.65 (dd, J=8.7, 2.5Hz, 1H), 7.50 (d, J=8.7Hz, 1H), 6.99 (d, J=10.0Hz, 2H), 4.22 (t, J=6.4Hz, 2H), 2.91 (d, J=11.4Hz, 2H), 2.49–2.45 (m, 2H), 2.02–1.90 (m, 4H), 1.63(d, J＝12.8Hz, 2H), 1.41-1.33(m, 1H), 1.29-1.22(m, 3H), 0.93(d, J＝6.5Hz, 3H).MS(ESI)m /z 396.9([M+H] ⁺ )

Example 25, 2-(3,4-dichlorophenyl)-6-(3-(pyrrolidin-1-yl)propoxy)pyridazin-3(2H)-one (compound 25)

Replace phenylhydrazine hydrochloride with 3,4-dichlorophenylhydrazine hydrochloride, 1,2-dibromoethane with 1,3-dibromopropane, and morpholine with pyrrolidine, and prepare according to the method of Example 1 target compound.

¹ H NMR (600MHz, CDCl ₃ ) δ 7.85 (d, J=2.5Hz, 1H), 7.61 (dd, J=8.7, 2.5Hz, 1H), 7.46 (d, J=8.7Hz, 1H), 6.95 (q, J=9.8Hz, 2H), 4.21 (t, J=6.4Hz, 2H), 2.62–2.58 (m, 2H), 2.53 (dd, J=8.5, 3.3Hz, 4H), 2.01–1.94 ( m,2H),1.81–1.74(m,4H).

MS(ESI) m/z 368.6([M+H] ⁺ )

Example 26, 2-(3,4-dichlorophenyl)-6-(3-(4-oxopiperidin-1-yl)propoxy)pyridazin-3(2H)-one (Compound 26 )

Replace phenylhydrazine hydrochloride with 3,4-dichlorophenylhydrazine hydrochloride, 1,2-dibromoethane with 1,3-dibromopropane, and morpholine with 4-piperidone, as in Example 1 method to prepare the target compound.

¹ H NMR (600MHz, CDCl ₃ ) δ 7.87 (d, J=2.4Hz, 1H), 7.64 (dd, J=8.7, 2.5Hz, 1H), 7.49 (d, J=7.4Hz, 1H), 7.02 –6.96(m, 2H), 4.26(t, J=6.3Hz, 2H), 2.78(dd, J=13.4, 7.4Hz, 4H), 2.62(t, J=7.2Hz, 2H), 2.45(t, J=6.0Hz,4H),2.05–1.96(m,2H).MS(ESI)m/z 396.5([M+H] ⁺ )

Example 27, 2-(3,4-dichlorophenyl)-6-(3-(4-ethylpiperazin-1-yl)propoxy)pyridazin-3(2H)-one (Compound 27 )

Replace phenylhydrazine hydrochloride with 3,4-dichlorophenylhydrazine hydrochloride, 1,2-dibromoethane with 1,3-dibromopropane, and morpholine with N-ethylpiperazine, as per example 1 to prepare the target compound.

¹ H NMR (600MHz, CDCl ₃ ) δ 7.84 (d, J=2.4Hz, 1H), 7.61 (dd, J=8.7, 2.5Hz, 1H), 7.47 (d, J=8.7Hz, 1H), 7.00 –6.94 (m, 2H), 4.19 (t, J=6.4Hz, 2H), 2.73–2.41 (m, 12H), 1.98–1.88 (m, 2H), 1.12 (t, J=7.2Hz, 3H). MS(ESI)m/z 411.4([M+H] ⁺ )

Example 28, 2-(3,4-dichlorophenyl)-6-(3-(4-methyl-1,4-diazeptan-1-yl)propoxy)pyridazine-3 (2H)-ketone (Compound 28)

Replace phenylhydrazine hydrochloride with 3,4-dichlorophenylhydrazine hydrochloride, 1,2-dibromoethane with 1,3-dibromopropane, and morpholine with N-methylhomopiperazine. The title compound was prepared by the method of Example 1.

¹ H NMR (600MHz, CDCl ₃ ) δ 7.82 (d, J=2.4Hz, 1H), 7.59 (dd, J=8.7, 2.4Hz, 1H), 7.47 (d, J=8.7Hz, 1H), 7.01 –6.95(m, 2H), 4.17(t, J=6.1Hz, 2H), 3.33–3.22(m, 4H), 3.06(d, J=3.9Hz, 2H), 2.87(t, J=6.1Hz, 2H),2.78–2.73(m,4H),2.23–2.17(m,2H),1.99–1.91(m,3H).MS(ESI)m/z 411.6([M+H] ⁺ )

Example 29, 2-(3,4-dichlorophenyl)-6-(3-(3,5-dimethylpiperidin-1-yl)propoxy)pyridazin-3(2H)-one (Compound 29)

Change phenylhydrazine hydrochloride to 3,4-dichlorophenylhydrazine hydrochloride, 1,2-dibromoethane to 1,3-dibromopropane, morpholine to 3,5-dimethylpiperidine, The target compound was prepared according to the method of Example 1.

¹ H NMR (600MHz, CDCl ₃ ) δ 7.87 (t, J=2.9Hz, 1H), 7.65-7.61 (m, 1H), 7.48 (d, J=8.7Hz, 1H), 7.02-6.94 (m, 2H), 4.27–4.17 (m, 2H), 2.88 (d, J=10.0Hz, 2H), 2.53–2.48 (m, 2H), 2.06–1.98 (m, 2H), 1.77–1.65 (m, 2H) ,1.50(t,J＝11.0Hz,2H),0.85(d,J＝6.5Hz,6H),0.54(q,J＝12.3Hz,2H).MS(ESI)m/z 410.6([M+H ] ⁺ )

Example 30, tert-butyl 4-(3-((1-(3,4-dichlorophenyl)-6-oxo-1,6-dihydropyridazin-3-yl)oxy)propoxy ) piperazine-1-carboxylate (compound 30)

Replace phenylhydrazine hydrochloride with 3,4-dichlorophenylhydrazine hydrochloride, 1,2-dibromoethane with 1,3-dibromopropane, and morpholine with N-Boc piperazine, as in Example 1 method to prepare the target compound.

¹ H NMR (600MHz, CDCl ₃ ) δ 7.79 (d, J=2.4Hz, 1H), 7.55 (dd, J=8.7, 2.4Hz, 1H), 7.38 (dd, J=8.7, 2.4Hz, 1H) ,6.92-6.85(m,2H),4.13(t,J＝6.3Hz,2H),3.39-3.28(m,4H),2.40(t,J＝7.2Hz,2H),2.30(s,4H), 1.90–1.81(m, 2H), 1.36(s, 9H).

MS(ESI) m/z 483.7([M+H] ⁺ )

Example 31, 2-(3,4-dichlorophenyl)-6-(3-(4-phenylpiperazin-1-yl)propoxy)pyridazin-3(2H)-one (Compound 31 )

Replace phenylhydrazine hydrochloride with 3,4-dichlorophenylhydrazine hydrochloride, 1,2-dibromoethane with 1,3-dibromopropane, and morpholine with 4-phenylpiperazine, as per example 1 to prepare the target compound.

¹ H NMR (600MHz, CDCl ₃ ) δ 7.82 (d, J=2.4Hz, 1H), 7.57 (dd, J=8.8, 2.4Hz, 1H), 7.38 (d, J=8.8Hz, 1H), 6.89 (q, J=9.7Hz, 2H), 4.13 (t, J=6.4Hz, 2H), 2.52–2.43 (m, 6H), 1.86–1.78 (m, 2H), 0.93 (t, J=7.2Hz, 6H).MS(ESI)m/z 370.9([M+H] ⁺ )

Example 32, 2-(3,4-dichlorophenyl)-6-(3-(dimethylamino)propoxy)pyridazin-3(2H)-one (32)

Replace phenylhydrazine hydrochloride with 3,4-dichlorophenylhydrazine hydrochloride, 1,2-dibromoethane with 1,3-dibromopropane, and morpholine with dimethylamine, as in Example 1 Preparation of the target compound.

¹ H NMR (600MHz, CDCl ₃ ) δ 7.82 (d, J=2.4Hz, 1H), 7.57 (dd, J=8.8, 2.4Hz, 1H), 7.38 (d, J=8.8Hz, 1H), 6.89 (q, J=9.7Hz, 2H), 4.13 (t, J=6.4Hz, 2H), 2.50 (m, 2H), 1.86–1.78 (m, 2H), 0.93 (t, J=7.2Hz, 6H) .MS(ESI)m/z 342.6([M+H] ⁺ )

Example 33, 2-(3,4-dichlorophenyl)-6-(3-(diethylamino)propoxy)pyridazin-3(2H)-one (compound 33)

Replace phenylhydrazine hydrochloride with 3,4-dichlorophenylhydrazine hydrochloride, 1,2-dibromoethane with 1,3-dibromopropane, and morpholine with diethylamine, as in Example 1 Preparation of the target compound.

¹ H NMR (600MHz, CDCl ₃ ) δ 7.82 (d, J=2.4Hz, 1H), 7.57 (dd, J=8.8, 2.4Hz, 1H), 7.38 (d, J=8.8Hz, 1H), 6.89 (q, J=9.7Hz, 2H), 4.13 (t, J=6.4Hz, 2H), 2.52–2.43 (m, 6H), 1.86–1.78 (m, 2H), 0.93 (t, J=7.2Hz, 6H).MS(ESI)m/z 370.5([M+H] ⁺ )

Example 34, 2-(3,4-dichlorophenyl)-6-(3-(dipropylamino)propoxy)pyridazin-3(2H)-one (compound 34)

Replace phenylhydrazine hydrochloride with 3,4-dichlorophenylhydrazine hydrochloride, 1,2-dibromoethane with 1,3-dibromopropane, and morpholine with dipropylamine, and prepare according to the method of Example 1 target compound.

¹ H NMR (600MHz, CDCl ₃ ) δ 7.90 (d, J=2.5Hz, 1H), 7.67 (dd, J=8.7, 2.5Hz, 1H), 7.51 (d, J=8.7Hz, 1H), 7.04 –6.97(m,2H),4.24(t,J=6.4Hz,2H),2.57(t,J=7.1Hz,2H),2.42–2.35(m,4H),1.95–1.85(m,2H), 1.51–1.41(m,4H),0.88(t,J=7.4Hz,6H).MS(ESI)m/z398.6([M+H] ⁺ )

Example 35, 2-(3,4-dichlorophenyl)-6-(4-morpholinobutoxy)pyridazin-3(2H)-one (compound 35)

Substitute phenylhydrazine hydrochloride with 3,4-dichlorophenylhydrazine hydrochloride and 1,2-dibromoethane with 1,4-dibromobutane, and prepare the target compound according to the method of Example 1.

¹ H NMR (600MHz, CDCl ₃ ) δ 7.75 (d, J=2.3Hz, 1H), 7.50 (dd, J=8.8, 2.3Hz, 1H), 7.30 (d, J=8.8Hz, 1H), 6.80 (dt, J=9.8, 8.3Hz, 2H), 4.02 (t, J=6.4Hz, 2H), 3.54–3.48 (m, 4H), 2.22 (dd, J=20.1, 12.6Hz, 6H), 1.66– 1.59(m,2H),1.50–1.43(m,2H).MS(ESI)m/z 398.3([M+H] ⁺ )

Example 36, 2-(3,4-dichlorophenyl)-6-(4-(4-methylpiperidin-1-yl)butoxy)pyridazin-3(2H)-one (Compound 36 )

Replace phenylhydrazine hydrochloride with 3,4-dichlorophenylhydrazine hydrochloride, 1,2-dibromoethane with 1,4-dibromobutane, and morpholine with 4-methylpiperidine. The title compound was prepared by the method of Example 1.

¹ H NMR (600MHz, CDCl ₃ ) δ 7.89 (d, J=2.4Hz, 1H), 7.65 (dd, J=8.7, 2.4Hz, 1H), 7.51 (d, J=8.7Hz, 1H), 7.01 –6.97(m,2H),4.19(t,J=6.4Hz,2H),2.91(d,J=11.4Hz,2H),2.41–2.32(m,2H),1.93(dd,J=20.6,8.6 Hz, 2H), 1.82–1.75 (m, 2H), 1.71–1.57 (m, 4H), 1.37 (ddd, J=14.6, 10.5, 5.2Hz, 1H), 1.30–1.22 (m, 2H), 0.93 ( d,J=6.5Hz,3H).MS(ESI)m/z 410.4([M+H] ⁺ )

Example 37, 2-(3,4-dichlorophenyl)-6-(4-(piperidin-1-yl)butoxy)pyridazin-3(2H)-one (compound 37)

Replace phenylhydrazine hydrochloride with 3,4-dichlorophenylhydrazine hydrochloride, 1,2-dibromoethane with 1,4-dibromobutane, and morpholine with piperidine, as in Example 1 Preparation of the target compound.

¹ H NMR (600MHz, CDCl ₃ ) δ 7.90-7.88 (m, 1H), 7.68-7.66 (m, 1H), 7.53 (dd, J=8.7, 4.8Hz, 1H), 7.08-6.97 (m, 2H) ).4.25–4.21(m,2H), 3.15–3.06(m,2H), 2.97–2.88(m,2H), 2.78–2.70(m,2H), 1.98–1.53(m,10H).MS(ESI) )m/z 396.7([M+H] ⁺ )

Example 38, 6-(3-(4-methylpiperidin-1-yl)propoxy)-2-(p-tolyl)pyridazin-3(2H)-one (compound 38)

Replace phenylhydrazine hydrochloride with p-toluenehydrazine hydrochloride, replace 1,2-dibromoethane with 1,3-dibromopropane, and replace morpholine with 4-methylpiperidine, and prepare according to the method of Example 1 target compound.

¹ H NMR (600 MHz, CDCl ₃ ) δ 7.52 (d, J=8.3 Hz, 2H), 7.23 (d, J=8.2 Hz, 2H), 6.95 (q, J=9.7 Hz, 2H), 4.18 (t , J=6.4Hz, 2H), 2.88 (d, J=11.3Hz, 2H), 2.45–2.42 (m, 2H), 2.37 (s, 3H), 1.97–1.86 (m, 4H), 1.60 (d, J=12.8Hz, 2H), 1.39–1.31 (m, 1H), 1.28–1.18 (m, 2H), 0.91 (d, J=6.5Hz, 3H). MS (ESI) m/z 342.7 ([M+ H] ⁺ )

Example 39, 6-(3-(piperidin-1-yl)propoxy)-2-(p-tolyl)pyridazin-3(2H)-one (compound 39)

Substitute phenylhydrazine hydrochloride with p-toluenehydrazine hydrochloride, 1,2-dibromoethane with 1,3-dibromopropane, and morpholine with piperidine, and prepare the target compound according to the method of Example 1.

¹ H NMR (600 MHz, CDCl ₃ ) δ 7.48 (d, J=8.3 Hz, 2H), 7.22 (d, J=8.1 Hz, 2H), 6.98-6.93 (m, 2H), 4.17 (t, J= 6.2Hz, 2H), 2.59(dd, J=18.2, 10.4Hz, 6H), 2.35(s, 3H), 2.09-1.97(m, 2H), 1.71-1.61(m, 4H), 1.46(s, 2H) ).MS(ESI)m/z 328.7([M+H] ⁺ )

Example 40, 2-(4-fluorophenyl)-6-(3-(4-methylpiperidin-1-yl)propoxy)pyridazin-3(2H)-one (Compound 40)

Replace phenylhydrazine hydrochloride with p-fluorophenylhydrazine hydrochloride, 1,2-dibromoethane with 1,3-dibromopropane, and morpholine with 4-methylpiperidine, according to the method of Example 1 Preparation of the target compound.

¹ H NMR (600 MHz, CDCl ₃ ) δ 7.68-7.63 (m, 2H), 7.15-7.09 (m, 2H), 7.01-6.95 (m, 2H), 4.19 (t, J=6.4Hz, 2H), 2.88(d,J＝11.1Hz,2H),2.48-2.40(m,2H),1.98-1.84(m,4H),1.61(d,J＝12.8Hz,2H),1.35(d,J＝6.0Hz ,1H),1.23(q,J=11.4Hz,2H),0.91(d,J=6.5Hz,3H).MS(ESI)m/z 346.5([M+H] ⁺ )

Example 41, 2-(4-Fluorophenyl)-6-(3-(piperidin-1-yl)propoxy)pyridazin-3(2H)-one (Compound 41)

Substitute phenylhydrazine hydrochloride with p-fluorophenylhydrazine hydrochloride, 1,2-dibromoethane with 1,3-dibromopropane, and morpholine with piperidine, and prepare the target compound according to the method of Example 1.

¹ H NMR (600 MHz, CDCl ₃ ) δ 7.65-7.61 (m, 2H), 7.14-7.07 (m, 2H), 6.99 (d, J=9.7 Hz, 2H), 4.20 (t, J=6.2 Hz, 2H), 2.66(dd, J＝17.3, 9.3Hz, 6H), 2.15-2.00(m, 2H), 1.75-1.68(m, 4H), 1.50(s, 2H). MS(ESI) m/z 332.5 ([M+H] ⁺ )

Example 42, 2-(4-chlorophenyl)-6-(3-(piperidin-1-yl)propoxy)pyridazin-3(2H)-one (compound 42)

Substitute phenylhydrazine hydrochloride with p-chlorophenylhydrazine hydrochloride, 1,2-dibromoethane with 1,3-dibromopropane, and morpholine with piperidine, and prepare the target compound according to the method of Example 1.

¹ H NMR (600 MHz, CDCl ₃ ) δ 7.68-7.64 (m, 2H), 7.42-7.38 (m, 2H), 6.97 (s, 2H), 4.20 (t, J=6.4Hz, 2H), 2.42 ( dd,J=22.1,14.4Hz,6H),2.04–1.86(m,2H),1.65–1.53(m,4H),1.43(s,2H).MS(ESI)m/z348.7([M+ H] ⁺ )

Example 43, 2-(4-chlorophenyl)-6-(3-(4-methylpiperidin-1-yl)propoxy)pyridazin-3(2H)-one (compound 43)

Replace phenylhydrazine hydrochloride with p-chlorophenylhydrazine hydrochloride, 1,2-dibromoethane with 1,3-dibromopropane, and morpholine with 4-methylpiperidine, according to the method of Example 1 Preparation of the target compound.

¹ H NMR (600 MHz, CDCl ₃ ) δ 7.68-7.65 (m, 2H), 7.43-7.39 (m, 2H), 7.01-6.96 (m, 2H), 4.21 (t, J=6.4Hz, 2H), 2.89(d,J＝11.4Hz,2H),2.49-2.43(m,2H),1.99-1.87(m,4H),1.62(d,J＝12.9Hz,2H),1.42-1.33(m,1H) ,1.28-1.21(m,3.6Hz,2H),0.92(d,J=6.5Hz,3H).MS(ESI)m/z 362.8([M+H] ⁺ )

Example 44, 6-(3-(piperidin-1-yl)propoxy)-2-(naphthalen-2-yl)pyridazin-3(2H)-one (compound 44)

Substitute phenylhydrazine hydrochloride with 2-naphthylhydrazine hydrochloride, 1,2-dibromoethane with 1,3-dibromopropane, and morpholine with piperidine, and prepare the target compound according to the method of Example 1.

¹ H NMR (600 MHz, CDCl ₃ ) δ 8.20 (d, J=1.7 Hz, 1H), 7.95-7.85 (m, 3H), 7.79 (dd, J=8.8, 2.1 Hz, 1H), 7.54-7.49 ( m, 2H), 7.03 (dd, J=22.4, 9.7Hz, 2H), 4.25 (t, J=6.4Hz, 2H), 2.45 (dd, J=24.5, 16.7Hz, 6H), 2.05–1.92 (m ,2H),1.66–1.55(m,4H),1.45(s,2H).MS(ESI)m/z 364.5([M+H] ⁺ )

Example 45, 6-(3-(4-methylpiperidin-1-yl)propoxy)-2-(naphthalen-2-yl)pyridazin-3(2H)-one (compound 45)

Replace phenylhydrazine hydrochloride with 2-naphthylhydrazine hydrochloride, 1,2-dibromoethane with 1,3-dibromopropane, and morpholine with 4-methylpiperidine, according to the method of Example 1 Preparation of the target compound.

¹ H NMR (600 MHz, CDCl ₃ ) δ 8.20 (d, J=1.8 Hz, 1H), 7.89 (ddd, J=16.8, 11.9, 6.1 Hz, 3H), 7.79 (dd, J=8.8, 2.1 Hz, 1H), 7.54–7.47(m, 2H), 7.03(dd, J=23.3, 9.7Hz, 2H), 4.25(t, J=6.4Hz, 2H), 2.90(d, J=11.4Hz, 2H), 2.53–2.43 (m, 2H), 2.01–1.89 (m, 4H), 1.62 (d, J=12.8Hz, 2H), 1.43–1.18 (m, 3H), 0.93 (d, J=6.5Hz, 3H) .MS(ESI)m/z 378.8([M+H] ⁺ )

Example 46, 2-cyclopentyl-6-(3-(piperidin-1-yl)propoxy)pyridazin-3(2H)-one (compound 46)

Substitute phenylhydrazine hydrochloride with cyclopentylhydrazine hydrochloride, 1,2-dibromoethane with 1,3-dibromopropane, and morpholine with piperidine, and prepare the target compound according to the method of Example 1.

¹ H NMR (600MHz, CDCl ₃ ) δ 6.67-6.50 (m, 2H), 5.19-5.00 (m, 1H), 3.93 (dd, J=14.8, 8.5Hz, 2H), 2.19 (dd, J=22.9 ,15.5Hz,6H),1.82–1.51(m,8H),1.35(dd,J=23.9,18.4Hz,6H),1.18(s,2H).MS(ESI)m/z 306.5([M+H ] ⁺ )

Example 47, 6-(4-(dimethylamino)butoxy)-2-phenylpyridazin-3(2H)-one (compound 47)

Substitute 1,2-dibromoethane with 1,4-dibromobutane and morpholine with dimethylamine, and prepare the target compound according to the method of Example 1.

Table 1. Preferred compound numbers and their structural formulas prepared in the examples

Pharmacological Example

In the following embodiment, the adopted homogenate includes A homogenate and B homogenate, and the configuration methods are as follows:

A homogenate contains Tris-HCl buffer at a final concentration of 0.01 M and a sucrose solution at a final concentration of 0.32 M, pH 7.4.

B homogenate was 0.01M Tris-HCl buffer, pH 7.4.

The C homogenate was 50 mM Tris buffer, pH 7.4.

Example 48. Preparation of σ ₁ receptor membrane and determination of ligand affinity (Ki value)

Preparation of σ ₁ Receptor Membranes

After the guinea pig was decapitated, the brain tissue was quickly taken out on ice, and the obtained brain tissue was added to a centrifuge tube. To this centrifuge tube, 5 ml of A homogenate was added and homogenized. Then continue to add A homogenate to the centrifuge tube to reach 10ml A homogenate/g brain tissue. The homogenized centrifuge tube was centrifuged at 1000 rpm for 10 min. After centrifugation, take the supernatant, add A homogenate to the supernatant to adjust to 2ml/g, centrifuge, take the supernatant, centrifuge at 1000rpm for 10min at 4°C, take the supernatant, centrifuge at 11500rpm for 25min at 4°C . Take the precipitate, add A homogenate to the obtained precipitate to adjust to 3ml A homogenate/g precipitate, incubate at 25°C for 15min, then centrifuge at 11500rpm for 25min at 4°C, and take the precipitate as the σ ₁ acceptor membrane.

Receptor binding assay materials

Isotope ligand [ ³ H]-(+)-pentazocine (250μCi, NET-1056250UC), purchased from Perkin--Elmer Company;

Haloperidol was purchased from Sigma-Aldrich;

GF/C glass fiber filter paper, purchased from Whatman Company;

PPO, POPOP and lipid-soluble scintillation fluid were purchased from Shanghai No. 1 Reagent Factory.

laboratory apparatus

Wallace 1450 MicroBeta TriLux Scintillation Counter, Perkin Elmer Company

experimental method

1. Bradford method for quantitative determination of protein

Refer to the instructions of commercially available kits.

2. Receptor saturation binding experiment.

(1) the obtained σ ₁ receptor membrane is mixed with homogenate B and dispersed evenly with a homogenizer, and the suspension of B homogenate reference protein determination quantitative membrane is continued to be added for subsequent use;

(2) 100 μL of membrane preparation was added to each reaction tube;

(3) Add 100 μL of homogenate B to the total binding tube (TB), and add 100 μL of haloperidol (final concentration 10 ^-5 M) to the non-specific binding tube (NB);

(4) 10 μL of the radioligand [3H]-(+)-tebuzocine was added to each reaction tube, and the final concentrations were 32.00, 16.00, 8.00, 4.00, 2.00, 1.00, 0.50, 0.25nM;

(5) Incubate each reaction tube at 25°C for 3 hours. After the reaction is completed, quickly filter the ligands bound in each reaction tube under reduced pressure, and wash thoroughly with ice-cold test buffer, then take out the filter and put it in a 2ml scintillation cup, add 1ml toluene scintillation fluid and mix well;

(6) Put the scintillation vial into the liquid scintillation counter for counting.

3. Competitive binding assay of σ ₁ receptor

(1) First, the σ ₁ receptor membrane prepared above was mixed with B homogenate and dispersed evenly with a homogenizer, and B homogenate was added to form a 50ml suspension for subsequent use;

(2) 100 μL of the membrane preparation (that is, the above-mentioned suspension) was added to each reaction tube;

(3) Add 100 μL of B solution to the total binding tube (TB), add 100 μL of haloperidol (final concentration 10 ^-5 M) to the non-specific binding tube (NB), and add 100 μL of the test compound to the specific binding tube (SB) of each test compound. Test compound (final concentration 10 ^-5 M);

(4) 10 μL of the radioligand [3H]-(+)-pentazocine (final concentration 4nM) was added to each reaction tube, and each test compound was the compound prepared in Examples 1-47;

(5) Incubate the above reaction tubes at 25°C for 3 hours, and after the reaction is completed, quickly filter the ligands in the reaction tubes under reduced pressure (wherein Whatman test paper uses 0.25% PEI (polyetherimide) 2 hours in advance (Polyetherimide) solution saturated), wash thoroughly with ice-cold assay buffer, then take out the filter and put it into a 2ml scintillation cup, add 1ml of toluene scintillation fluid and mix well;

(6) Put the scintillation vial into the liquid scintillation counter for counting.

4. Data Statistical Processing

First, the inhibition rate of each test compound was calculated by the following formula:

Inhibition rate (I%)=(TB-SB)/(TB-NB)×100%,

in,

TB: total binding constant;

NB: nonspecific binding constant;

SB: Binding constant of the compound.

Next, the _IC50 of each test compound was calculated by the logit method.

Then, the K _d value and Bmax of each radioligand were obtained by Scatchard plotting;

Finally, the _Ki value of the tested compound was determined by the following formula:

Ki=IC ₅₀ /(1+C/K _d ),

Among them, C in this formula is the concentration of free isotope.

Some results are shown in Table 2.

Example 49. Preparation of σ ₂ receptor membrane and determination of ligand affinity (Ki value)

Preparation of σ ₂ Receptor Membranes

After the guinea pig was decapitated, the brain tissue was quickly removed by operation on ice, and the obtained brain tissue was added to a centrifuge tube. Add 0.01M Tris HCl and 0.32M sucrose solution to the centrifuge tube, then homogenize in 4th gear for 3-4s, homogenize 4 times, and then continue to add 0.01M Tris HCl and 0.32M sucrose solution to adjust to 10ml/g, Adjust the weight of the homogenized test tube with a balance, then centrifuge at 1000r for 10min; take the supernatant, add 0.01M Tris HCl and 0.32M sucrose solution to it, adjust it to 2ml/g, and then centrifuge at 1000r, 4°C for 10min ; Take the supernatant and centrifuge at 11000r for 30min at 4°C; take the precipitate, suspend it with 0.01M Tris HCl and 0.32M sucrose solution for 30s, adjust to 3ml/g, then incubate at 25°C for 15min, and then Centrifuge at 11000g for 30min; take the supernatant, store at -20°C for more than 12h, incubate with 50Mm-Tris during use, then centrifuge, and take the precipitate as the σ ₂ receptor membrane.

Receptor binding assay materials

The isotope ligand [ ³ H]-DTG ([ ³ H]-DTG, 250 μCi, NET-986250UC), purchased from Perkin--Elmer Company; DTG) is di-o-tolylguanidine.

DTG was purchased from Sigma-Aldrich;

(+)-SKF 10047 was purchased from Sigma-Aldrich;

GF/C glass fiber filter paper, purchased from Whatman Company;

PPO, POPOP and lipid-soluble scintillation fluid were purchased from Shanghai No. 1 Reagent Factory.

laboratory apparatus

Wallace 1450 MicroBeta TriLux Scintillation Counter, Perkin Elmer Company

experimental method

1. Bradford method for quantitative determination of protein

Refer to the instructions of commercially available kits.

2. Competitive binding experiment of sigma-2 receptor.

(1) first, the obtained σ ₂ receptor membrane is mixed with C homogenate (being 50mM Tris buffer, pH value is 7.4) and uniformly dispersed with a homogenizer to obtain a membrane preparation for subsequent use;

(2) 100 μL of membrane preparation and 100 μL of C homogenate were added to each reaction tube;

(3) Add 100 μL of C homogenate to the total binding tube (TB), add 5uM DTG 100 μL (final concentration 0.5*10 ⁻⁵ M) to the non-specific binding tube (NB), and add 100 μL of each test compound to the specific binding tube (SB) Test compound (final concentration 10 ^{- 5} M); 100nM (+)-NANM (CAS number: 133005-41-1) shields sigma-1 receptor; each test compound is the compound prepared in Example 1-47;

(4) Add 10 μL of radioligand 3H-DTG (final concentration 5nM) to each reaction tube (two parallel tubes are set for each reaction tube, and each tube is placed on ice when adding samples);

(5) Incubate each reaction tube at 25°C for 120 min. After the reaction is completed, quickly filter the ligands bound in each reaction tube under reduced pressure (whatman test paper is soaked in 0.5% PEI), and use ice-cold test Wash the buffer well, then take out the filter and put it into a 2ml scintillation cup, add 1ml of toluene scintillation fluid and mix well;

(6) Put the scintillation vial into the liquid scintillation counter for counting.

3. Data Statistical Processing

First, the inhibition rate of each test compound was calculated by the following formula:

Inhibition rate (I%)=(TB-SB)/(TB-NB)×100%,

in,

TB: total binding constant;

NB: nonspecific binding constant;

SB: Binding constant of the compound.

Next, use the logit method to calculate the IC ₅₀ of each test compound;

Then, the K _d value and Bmax of each radioligand were obtained by Scatchard plotting;

Finally, the _Ki value of the tested compound was determined by the following formula:

Ki=IC ₅₀ /(1+C/K _d ),

Some results are shown in Table 2.

Example 50. Determination of functionality of ligands for σ ₁ receptors

Preparation of σ ₁ Receptor Membranes

According to the method of "Preparation of σ ₁ Receptor Membrane" in Example 48, a σ ₁ receptor membrane was prepared.

Receptor binding assay materials

Isotope ligand [ ³ H]-(+)-pentazocine (250μCi, NET-1056250UC), purchased from Perkin--Elmer Company;

Haloperidol and phenytoin were purchased from Sigma-Aldrich;

GF/C glass fiber filter paper, purchased from Whatman Company;

PPO, POPOP and lipid-soluble scintillation fluid were purchased from Shanghai No. 1 Reagent Factory;

laboratory apparatus

Wallace 1450 MicroBeta TriLux Scintillation Counter, Perkin Elmer Company

experimental method

1. Bradford method for quantitative determination of protein

Refer to the instructions of commercially available kits.

2. σ ₁ receptor functional experiment

(1) firstly, the obtained σ ₁ receptor membrane was mixed with homogenate B and dispersed evenly with a homogenizer, and the homogenate B was continuously added to form a 50ml suspension, for subsequent use;

(1) 100 μL of the membrane preparation (that is, the above suspension) was added to each reaction tube;

(2) Add 100 μL of B solution to the total binding tube (TB), and add 100 μL of haloperidol to the non-specific binding tube (NB) (final concentration 10 ^-5 M);

(3) Add 100 μL of the test compound (final concentration 10 ^-5 M) to each test compound-specific binding tube (SB), and each test compound is the compound prepared in Examples 1-47;

(4) Add 10 μL of radioligand [3H]-(+)-pentazocine (final concentration 4nM) to each reaction tube;

(5) Incubate the above reaction tubes at 25°C for 3 hours. After the reaction is completed, quickly filter the ligands bound in each reaction tube under reduced pressure (where Whatman test paper is saturated with 0.25% PEI solution 2 hours in advance), and cool with ice. The test buffer was fully washed, and then the filter was taken out and placed in a 2ml scintillation cup, and 1ml of toluene scintillation fluid was added and mixed;

(6) Put the scintillation vial into the liquid scintillation counter for counting.

3. Data Statistical Processing

According to the method of "statistical processing of data" in Example 48, the K _i value of each test compound was calculated.

Among them, the functional experiment of the σ ₁ receptor is judged by detecting the change of the receptor affinity of the σ ₁ receptor allosteric agent phenytoin to the tested compound. Phenytoin has little effect on σ ₁ receptor antagonists, or weakly weakens the compound's affinity for receptors, but can significantly increase the affinity between σ ₁ receptor agonists and receptors. The σ ₁ receptor functionality of the tested compounds can be judged by comparing the changes in the σ ₁ receptor affinity (Ki value) of the tested compounds with and without phenytoin.

Table 2. Affinities (Ki values) of some compounds for σ ₁ receptor and σ ₂ receptor

Example 51. Acute toxicity study

The compounds prepared in Examples 1-47 were subjected to acute toxicity experiments, as follows:

Sequential method limit experiment

ICR mice, half male and half male, were randomly divided into several groups, 2-5 mice in each group, respectively the compound 2000 mg/kg group and the solvent group (at the same time, positive control groups were set: gabapentin group, SIRA group), press 0.2 ml /10g administered by gavage. Animals were observed for death within 3 days.

Among them, if three or more animals survived within three days, and there is no obvious abnormality in their life status, continue to observe until the end of the experiment 7 days later. If three or more animals died within three days, the median lethal dose method was used to determine its LD50.

median lethal dose test

ICR mice, half male and half male, were randomly divided into several groups, 4 mice in each group, respectively 1500mg/kg, 1000mg/kg, 500mg/kg groups and solvent groups of each compound (at the same time, positive control groups were set: gabapentin group and SIRA group). ), administered by gavage at 0.2ml/10g, and observed the death of animals within 1-3 days.

result

As a result, the LD ₅₀ of a single dose of mice was greater than 2000 mg/kg, which was comparable to that of the positive control drug S1RA (>2000 mg/kg), and had less acute toxicity. Some results are shown in Table 3.

Example 52. Formalin-induced pain model experiment in mice

The compounds prepared in Examples 1-47 were subjected to a formalin-induced pain model experiment in mice, as follows:

laboratory animals

Healthy ICR mice, male, 22-40g, were provided by Nanjing Qinglongshan Animal Breeding Center.

main reagent

Positive drugs tested: gabapentin, pregabalin, S1RA (E-52862)

Formaldehyde solution, 1002012, Longxi Chemical Industry;

Sodium chloride injection, H32026305, Xuzhou No. 5 Pharmaceutical Factory Co., Ltd.;

PEG400, 20111202, Will Chemical.

laboratory apparatus

stopwatch

observation glass

experimental method

ICR mice were taken and randomly divided into negative control group, model group, positive drug dose groups (gabapentin, pregabalin, S1RA) and compound dose groups (see Table 3 for specific doses), with 10 mice in each group. The negative control group and model group were given the corresponding solvent double distilled water by gavage, the positive drug group was given the corresponding positive drug by gavage, and the compound groups were given the corresponding dose of compound by gavage, and the gavage volume was 0.1ml/10g. After gavage for 15 minutes, 20 μL of 2.5% formalin was subcutaneously injected into the left hind paw of the mice to establish a model, and the formation of a skin mound was regarded as the successful standard of modeling. After successful modeling, the time of mice licking and biting the injection foot site was observed at 0-5 minutes and 15-45 minutes after modeling.

Data Statistical Processing

The experimental data were expressed as mean±standard deviation (Mean±SD), and the comparison was performed by one-way analysis of variance; then, the ED ₅₀ was calculated by the probability unit regression method, and some results are shown in Table 4.

Table 3. Effects of gabapentin, SIRA, compounds 23, 24 on the number of leg lifts in formalin-induced pain rats

Note: *P<0.05, **P<0.01VS model group.

Table 4. Results of in vivo animal model experiments for preferred compounds

The above in vitro receptor binding test shows that the compounds involved in the present invention have a high affinity for the σ ₁ receptor, but a low affinity for the σ ₂ receptor. Selective antagonism of σ ₁ receptors, indicating potential for analgesic activity.

In addition, animal test results also show that these compounds can significantly improve both formalin-induced phase I and phase II pain. Since these in vitro action targets and in vivo pharmacological models are closely related to the sigma ₁ receptor-mediated nervous system regulation response, especially pain, the compounds involved in the present invention have the potential to treat pain, especially neuralgia.

Formulation Example

Example 53, tablet preparation

Taking the compounds prepared in Examples 1-47 as active ingredients, and taking tablet dosage forms as an example, the pharmaceutical compositions of the present invention are prepared according to the following formula:

Pass the raw and auxiliary materials through an 80-mesh sieve for later use, weigh the active ingredients, microcrystalline cellulose, lactose, and povidone K30 in the formulation, add them to a high-speed mixing preparation machine, stir and mix at a low speed, add an appropriate amount of purified water, stir at a low speed, and mix at a high speed. Cut and granulate, then dry the wet granules at 60°C for 3 hours, sieve them with a 24-mesh sieve, and add the prescribed amounts of sodium starch glycolate, silicon dioxide and magnesium stearate, mix them together, and press on a rotary tablet machine to obtain Pharmaceutical compositions in tablet dosage form.

In the description of this specification, description with reference to the terms "one embodiment," "some embodiments," "example," "specific example," or "some examples", etc., mean specific features described in connection with the embodiment or example , structure, material or feature is included in at least one embodiment or example of the present invention. In this specification, schematic representations of the above terms are not necessarily directed to the same embodiment or example. Furthermore, the particular features, structures, materials or characteristics described may be combined in any suitable manner in any one or more embodiments or examples. Furthermore, those skilled in the art may combine and combine the different embodiments or examples described in this specification, as well as the features of the different embodiments or examples, without conflicting each other.

Although the embodiments of the present invention have been shown and described above, it should be understood that the above-mentioned embodiments are exemplary and should not be construed as limiting the present invention. Embodiments are subject to variations, modifications, substitutions and variations.

Claims (12)

1. a compound, it is the stereoisomer of compound shown in formula I or compound shown in formula I, tautomer or pharmaceutically acceptable salt

in, R ₁ is an optionally substituted aryl group containing 6-14 ring atoms, and the substituted substituent is selected from at least one of C _1-6 alkyl, cyano, hydroxyl and halogen; Z is -R _c -R _d -, R _c is O, and R _d is a straight-chain or branched alkylene group with 3 to 4 carbon atoms; Q together with its attached _Ra and _Rb form

R ₄ and R ₅ are independently one or more selected from hydrogen, C _1-5 alkyl, hydroxyl and tert-butoxycarbonyl; m is 0, 1 or 2; X is one of oxygen, nitrogen, or CH. 2. The compound according to claim 1, wherein R ₁ is optionally substituted phenyl or optionally substituted naphthyl, and the substituted substituent is selected from C _1-4 alkyl, cyano, at least one of hydroxyl and halogen. 3. The compound according to claim 1, wherein the halogen is fluorine, chlorine, bromine and iodine. 4. The compound of claim 1, wherein R ₁ is phenyl, dichlorophenyl, tolyl, fluorophenyl, chlorophenyl or naphthyl; Z is -OCH ₂ CH ₂ CH ₂ - or -OCH ₂ CH ₂ CH ₂ CH ₂ -; R ₄ and R ₅ are each independently one or more selected from hydrogen, methyl, ethyl, propyl, and tert-butoxycarbonyl. 5. A compound, characterized in that the compound is at least one of the following compounds, or a stereoisomer, tautomer or pharmaceutically acceptable salt of at least one of the following compounds:

6. A pharmaceutical composition, characterized by comprising the compound according to any one of claims 1 to 5. 7. The pharmaceutical composition according to claim 6, further comprising a pharmaceutically acceptable excipient, carrier, adjuvant or a combination thereof. 8. The pharmaceutical composition of claim 7, wherein the excipient comprises a vehicle. 9 . The pharmaceutical composition according to claim 6 , further comprising a drug for preventing or treating pain diseases different from the compound according to any one of claims 1 to 5 , which is different from claim 1 . 10 . The medicine for preventing or treating pain diseases of any one of the compounds of ~5 is at least one selected from the following: Non-steroidal anti-inflammatory painkillers, central painkillers, narcotic painkillers, traditional Chinese medicine compound painkillers. 10 . The pharmaceutical composition according to claim 9 , wherein the drug for preventing or treating pain diseases different from the compound according to any one of claims 1 to 5 is at least one selected from the following: 10 . : Aspirin, ibuprofen, indomethacin, paracetamol, phenylbutazone, rofecoxib, celecoxib, tramadol, fentanyl, morphine, delperidine, anisodamine, naproxen, fenbid, voltaren and analgesic new. 11. Use of the compound of any one of claims 1 to 5 or the pharmaceutical composition of any one of claims 6 to 10 in the preparation of a medicament for preventing or treating pain-related diseases. 12. The use according to claim 11, wherein the pain disease is neuralgia.