1',2'-dihydro-3'h-spiro[cyclobutane 1,4'-isoquinoline]-3'-one derivatives and their use

Abstract

This invention pertains to the medical field and relates to a salt of a 1',2'-dihydro-3'H-spiro[cyclobutane-1,4'-isoquinoline]-3'-one derivative and its applications. Specifically, it relates to the hydrochloride salt of the compound shown in general formula (A) below, or its cis-trans isomers, tautomers, enantiomers, diastereomers, meso compounds, racemates, or mixtures thereof. This compound can be used to prepare drugs for treating neuropsychiatric diseases.

Description

[0001] This application claims priority to the earlier application filed on May 12, 2021, with patent application number CN202110525528.0, entitled "Salts of 1′,2′-dihydro-3′H-spiro[cyclobutane 1,4′-isoquinoline]-3′-one derivatives and their applications thereto." The entire contents of the earlier application are incorporated herein by reference. Technical Field

[0002] This invention pertains to the pharmaceutical field, specifically relating to a salt of a novel 1′,2′-dihydro-3′H-spiro[cyclobutane-1,4′-isoquinoline]-3′-one derivative and its applications, particularly the hydrochloride salt, its preparation method, and compositions comprising the compound, as well as its applications in the pharmaceutical field. Technical Background

[0003] NMDAR (N-methyl-D-aspartate receptor) is an ionotropic glutamate receptor that primarily targets Ca2+. 2+ Ions are permeable and can be activated upon binding with glycine and glutamate, playing a crucial role in excitatory synaptic plasticity. Physiologically, they activate and trigger the opening of ion channels, generating an input current that only slowly inactivates. Under pathological conditions, they can lead to overactivation of NMDAR, a significant pathogenic mechanism of receptor excitotoxicity.

[0004] Physiological activity of NMDAR is essential for normal neurological function. Overactivation of NMDAR is associated with acute neurological diseases such as stroke or traumatic brain injury, and with chronic stress conditions such as neurodegenerative diseases. Many pathologies are thought to be associated with excessive NMDAR activity, and therefore NMDAR is potentially sensitive to NMDAR antagonists ([J]. Journal of Neurochemistry, 2006, 97(6): 1611-1626.).

[0005] Increasing evidence suggests the importance of NMDAR in inducing and maintaining central sensitization in pain states. Furthermore, NMDAR may also mediate peripheral sensitization and visceral pain ([J]. Nature, 2005, 438(7071):1162.). Extensive preclinical data support the potential of NMDAR antagonists to treat opioid-induced refractory pain, postoperative pain, and cancer pain. Other studies have shown that typical antidepressants alter the affinity of the NMDA receptor glycine site, and reduced NMDAR function contributes to antidepressant responses. Intravenous administration of a single subanesthetic dose of ketamine to patients with treatment-resistant depression significantly improved their condition, and the antidepressant effect lasted for one week ([J]. Archives of general psychiatry, 2006, 63(8):856-864.). Currently, S-ketamine nasal spray... It was approved for marketing in the United States in March 2019 for adjunctive treatment of resistant depression.

[0006] Based on their sites of action, NMDAR antagonists can be broadly classified into three categories: non-competitive (or allosteric) antagonists (ATD sites), such as afenidil, RGH-896, and EVT101; competitive antagonists (LBD sites), such as GLYX-13 and NRX-1074; and non-competitive antagonists, channel pore blockers (TMD sites), such as ketamine, dextromethorphan, and memantine. However, currently marketed NMDAR antagonists still commonly have various side effects, such as hallucinations, confusion, personality disorders, nightmares, agitation, attention deficit, mood changes, convulsions, and sedation ([J]. Biochemicalpharmacology, 2003, 66(6): 877-886.), thus limiting their use. The higher the affinity of the NMDAR channel pore antagonist, the slower its binding to NMDAR. The slower binding rate leads to Ca 2+ For sustained influx of NMDAR, very high drug concentrations are required to fully bind to the receptor, such as MK-801, which has a slow dissociation rate of NMDAR, resulting in significant psychogenic side effects.

[0007] Current research has confirmed that high-affinity, non-competitive NMDA receptor antagonists like MK-801, while able to prevent NMDAR activation and inhibit Ca2+ receptor activation, do not necessarily prevent NMDA receptor activation. 2+Intravenous influx, but its application is limited by significant psychomimetic adverse reactions. In contrast, low-affinity, non-competitive NMDAR antagonists (such as memantine) can reduce toxicity, thanks to the faster rate of NMDAR blockade and departure ([J]. European journal of pharmacology, 1996, 317(2-3): 377-381). Other studies have shown that the good clinical tolerability and symptomatic effect of memantine in the treatment of Alzheimer's disease are attributed to its moderate affinity for NMDA receptor channels and rapid dissociation from NMDAR ([J]. Neuropharmacology, 2009, 56(5): 866-875.); other studies have suggested that memantine has better inhibitory recovery kinetics than dizocilpine, which is considered to be the main determinant of the better clinical tolerability of memantine ([J]. ACS chemical neuroscience, 2018, 9(11): 2722-2730.). Therefore, the ability to rapidly dissociate from NMDAR is one of the key factors in the development of NMDAR antagonists.

[0008] CN106957285A discloses an aminocyclobutane derivative, specifically compound 1a1, which is an NMDAR antagonist and a potential treatment for depression and chronic pain. Its structure is shown below:

[0009]

[0010] Although there are NMDAR inhibitors with channel pore blockers (TMD sites) on the market, they have not yet achieved the effect of rapidly dissociating from NMDAR and reducing psychogenic side effects. Therefore, NMDAR inhibitors remain a research hotspot in the field of neuropsychiatry.

[0011] Patent application PCT / CN2020 / 129826 discloses a series of 1′,2′-dihydro-3′H-spiro[cyclobutane-1,4′-isoquinoline]-3′-one derivatives. In order to maximize the therapeutic effect of the above-mentioned active substances in clinical research, and to simplify product processing and improve product solubility in the pharmaceutical research and development process, this invention conducts a comprehensive and systematic study on the salts of the above-mentioned substances in order to obtain the most suitable salt form. Summary of the Invention

[0012] The present invention aims to provide a hydrochloride salt of 1′,2′-dihydro-3′H-spiro[cyclobutane 1,4′-isoquinoline]-3′-one derivatives. Compared with the free base form of the compound, the hydrochloride salt is beneficial to improve the bioavailability and solubility of the active substance, which is beneficial to subsequent clinical development and production development.

[0013] This invention provides a hydrochloride salt in the form of a compound represented by the following general formula (A), or a cis-trans isomer, tautomer, enantiomer, diastereomer, meso compound, racemic mixture, or mixture thereof.

[0014]

[0015] The present invention also provides the use of the hydrochloride salt of the compound represented by the above general formula (A) or in the form of its cis-trans isomers, tautomers, enantiomers, diastereomers, meso compounds, racemates, or mixtures thereof in the preparation of medicaments for treating neuropsychiatric diseases. Detailed Implementation

[0016] General terms and definitions

[0017] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. In case of any conflict, the definitions provided herein shall prevail. When trade names appear herein, they are intended to refer to the corresponding product or its active ingredient. All patents, published patent applications, and publications cited herein are incorporated herein by reference.

[0018] The terms “optional,” “optionally,” or “optionally exist” mean that the event or situation described below may, but is not necessarily, occur, and the description includes both the occurrence and non-occurrence of said event or situation.

[0019] The terms “selected from…”, “preferred…” and “more preferred…” refer to one or more elements from the group listed below, selected independently, and may include a combination of two or more elements. In this invention, one element from the group listed below is preferred.

[0020] The term "comprising" is an open-ended expression, meaning it includes the contents specified in this invention but does not exclude other aspects. It should be understood that the term "comprising" can also encompass a closed meaning, meaning "consisting of".

[0021] The term "substitution" means that one or more hydrogen atoms in a compound or group are replaced by other atoms or groups. This requires the formation of a stable valence state or compound. The expression "non-substitution" can also be understood as "unsubstituted." It should be understood that when the substituent is hydrogen, this can also mean that the corresponding group is "non-substituted" or "unsubstituted."

[0022] The term "hydrogen (H)" refers to a single hydrogen atom. Such a group can be attached to other groups, such as oxygen atoms, to form a hydroxyl group.

[0023] The term "halogen" refers to fluorine (F), chlorine (Cl), bromine (Br), or iodine (I).

[0024] In the context of this invention, the substituents of the disclosed compounds are disclosed according to the type or range of groups. Specifically, this invention includes every independent sub-combination of each member of these group types and ranges. For example, the term "alkyl" as used in this invention is preferably "C1-C6 alkyl"; the term "C1-C6 alkyl" specifically refers to the independently disclosed methyl, ethyl, C3 alkyl, C4 alkyl, C5 alkyl, and C6 alkyl groups. 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(CH3)2), n-butyl (n-Bu, -CH2CH2CH2CH3), isobutyl (i-Bu, -CH2CH(CH3)2), sec-butyl (s-Bu, -CH(CH3)CH2CH3), tert-butyl (t-Bu, - C(CH3)3), n-pentyl (-CH2CH2CH2CH2CH3), 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 2-Hexyl (-CH2CH(CH3)CH2CH3), n-Hexyl (-CH2CH2CH2CH2CH2CH3), 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(CH3)CH2CH(CH3)2), 3-Methyl-3-pentyl (-C(CH3)(CH2CH3)2), 2-Methyl-3-pentyl (-CH(CH2CH3)CH(CH3)2), 2,3-Dimethyl-2-butyl (-C(CH3)2CH(CH3)2), 3,3-Dimethyl-2-butyl (-CH(CH3)C(CH3)3), n-Heptyl, n-Octyl, etc.

[0025] A "subunit" is a group obtained by removing a hydrogen atom from a carbon atom that contains free valence electrons, and has two connection sites for attaching to the rest of the molecule. For example, "alkylene" or "alkyl subunit" refers to a saturated straight-chain or branched divalent hydrocarbon group.

[0026] The term "alkylene," used alone or in combination with other groups herein, refers to a straight-chain or branched saturated divalent hydrocarbon group. For example, the term "C1-C3 alkylene" refers to an alkylene having 1-3 carbon atoms, such as methylene, ethylene, or propylene. Parentheses in the structural formulas herein indicate the repetition of structural units. For example, n2 indicates the number of repetitions of the structural unit within the parentheses, where the structural unit within the parentheses is a methylene group substituted with one R6. When n2 is 1, 2, or 3, the structural fragment obtained by repeating the structural unit within the parentheses is a C1-C3 alkylene, and each -(CH2)- structure of the alkylene is substituted with one R6.

[0027] The term "alkenyl" refers to an unsaturated aliphatic hydrocarbon group consisting of a straight or branched chain of carbon and hydrogen atoms, having at least one double bond. The alkenyl groups described in this invention preferably have 2-8 carbon atoms, i.e., "C..." 2-8 "Alkenyl", for example, C 2-4 alkenyl, C 3-4 Alkenyl. Non-limiting examples of alkenyl include, but are not limited to, vinyl, allyl, (E)-2-methylvinyl, (Z)-2-methylvinyl, (E)-but-2-enyl, (Z)-but-2-enyl, (E)-but-1-enyl, (Z)-but-1-enyl, etc.

[0028] The term "alkynyl" refers to an unsaturated aliphatic hydrocarbon group consisting of a straight or branched chain of carbon and hydrogen atoms, having at least one triple bond. The alkynyl group described in this invention preferably has 2-8 carbon atoms, i.e., "C". 2-8 "Alkyne group", such as C 2-4 alkynyl group, C 3-4 Alkynyl. Non-limiting examples of alkynyl include, but are not limited to, ethynyl, prop-1-alkynyl, prop-2-alkynyl, but-1-alkynyl, but-2-alkynyl, but-3-alkynyl, etc.

[0029] The term "cycloalkyl" refers to a saturated cyclic hydrocarbon group consisting of carbon and hydrogen atoms, preferably containing one or two rings. The cycloalkyl group can be monocyclic, fused polycyclic, bridged, or spirocyclic. The cycloalkyl group preferably has 3-10 carbon atoms, i.e., "C3-C10 cycloalkyl," such as C3-C8, C3-C6, C5, C6, and C7 cycloalkyl. Non-limiting examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, bicyclic [2.2.1]heptyl, and spiro[3.3]heptyl. The term also covers cases where the carbon atom can be substituted with an oxygen (=O).

[0030] The term "alkoxy group" indicates that an alkyl group is attached to the remainder of the molecule via an oxygen atom, wherein the alkyl group has the meaning as described in this invention. Unless otherwise specified, the alkoxy group may contain 1-12 carbon atoms. According to one embodiment of the invention, the alkoxy group preferably contains 1-6 carbon atoms. According to another embodiment of the invention, the alkoxy group preferably contains 1-5 or 1-4 carbon atoms. According to yet another embodiment of the invention, the alkoxy group preferably contains 1-5 carbon atoms. The alkoxy group may optionally be substituted by one or more substituents described in this invention. Examples of alkoxy groups include, but are not limited to, methoxy (MeO, -OCH3), ethoxy (EtO, -OCH2CH3), 1-propoxy (n-PrO, n-propoxy, -OCH2CH2CH3), 2-propoxy (i-PrO, i-propoxy, -OCH(CH3)2), 1-butoxy (n-BuO, n-butoxy, -OCH2CH2CH2CH3), 2-methyl-l-propoxy (i-BuO, i-butoxy, -OCH2CH(CH3)2), 2-butoxy (s-BuO, s-butoxy, -OCH(CH3)CH2CH3), 2-methyl-2- Propoxy (t-BuO, t-butoxy, -OC(CH3)3), 1-pentoxy (n-pentoxy, -OCH2CH2CH2CH2CH3), 2-pentoxy (-OCH(CH3)CH2CH2CH3), 3-pentoxy (-OCH(CH2CH3)2), 2-methyl-2-butoxy (-OC(CH3)2CH2CH3), 3-methyl-2-butoxy (-OCH(CH3)CH(CH3)2), 3-methyl-l-butoxy (-OCH2CH2CH(CH3)2), 2-methyl-l-butoxy (-OCH2CH(CH3)CH2CH3), etc.

[0031] The term "thioalkyl" means that an alkyl group is attached to the remainder of the molecule by a sulfur atom, wherein the alkyl group has the meaning as described in this invention. Unless otherwise specified, the thioalkyl group preferably contains 1-12 carbon atoms. According to one embodiment of the invention, the thioalkyl group preferably contains 1-6 carbon atoms; more preferably, the thioalkyl group contains 1-5 or 1-4 carbon atoms. The thioalkyl group may optionally be substituted by one or more substituents described in this invention. Examples of thioalkyl groups include, but are not limited to, methylthio (MeS, -SCH3), ethylthio (EtS, -SCH2CH3), 1-propanethio (n-PrS, n-propanethio, -SCH2CH2CH3), 2-propanethio (i-PrS, i-propanethio, -SCH(CH3)2), 1-butanethio (n-BuS, n-butanethio, -SCH2CH2CH2CH3), 2-methyl-l-propanethio (i-BuS, i-butanethio, -SCH2CH(CH3)2), 2-butanethio (s-BuS, s-butanethio, -SCH(CH3)CH2CH3), 2-methyl-2- Propylthio (t-BuS, t-butylthio, -SC(CH3)3), 1-pentylthio (n-pentylthio, -SCH2CH2CH2CH2CH3), 2-pentylthio (-SCH(CH3)CH2CH2CH3), 3-pentylthio (-SCH(CH2CH3)2), 2-methyl-2-butylthio (-SC(CH3)2CH2CH3), 3-methyl-2-butylthio (-SCH(CH3)CH(CH3)2), 3-methyl-l-butylthio (-SCH2CH2CH(CH3)2), 2-methyl-l-butylthio (-SCH2CH(CH3)CH2CH3), etc.

[0032] The term "heterocyclic group" or "heterocyclic hydrocarbon group" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent containing 3 to 20 ring atoms, wherein one or more ring atoms are selected from nitrogen, oxygen, sulfur, and phosphorus, and the remaining ring atoms are carbon. Preferably, it contains 3 to 12 ring atoms, wherein 1 to 4 ring atoms are heteroatoms; most preferably, it contains 3 to 8 ring atoms, wherein 1 to 3 ring atoms are heteroatoms; most preferably, it contains 4 to 6 ring atoms, wherein 1 to 3 (e.g., 1, 2, and 3) ring atoms are heteroatoms. "Heterocyclic group" or "heterocyclic hydrocarbon group" is not aromatic. The term also covers cases where the C atom is substituted with an oxo (=O) and / or the S atom on the ring is substituted with one or two oxo (=O) atoms and / or the P atom on the ring is substituted with one or two oxo (=O) atoms.

[0033] Examples of heterocyclic groups include, but are not limited to, four-membered rings, such as aza-butyl and oxa-butyl rings; or five-membered rings, such as tetrahydrofuranyl, dioxolinyl, pyrrolyl, imidazoyl, pyrazolyl, pyrrolinyl, oxopyrrolyl, 2-oxoimidazolidin-1-yl; or six-membered rings, such as tetrahydropyranyl, piperidinyl, morpholinyl, dithiaalkyl, thiomorpholinyl, piperazine, 1,1-dioxo-1,2-thiazin-2-yl or trithiaalkyl; or seven-membered rings, such as diaza-butyl... The heterocyclic group may optionally be benzofused.

[0034] Heterocyclic groups can also be bicyclic, without limitation, such as five-membered fused five-membered rings, such as hexahydrocyclopentane[c]pyrrole-2(1H)-yl ring; or five-membered fused six-membered bicyclic rings, such as hexahydropyrrolo[1,2-a]pyrazine-2(1H)-yl ring.

[0035] As mentioned above, heterocycles can be unsaturated, meaning they can contain one or more double bonds without limitation. For example, unsaturated heterocycles containing nitrogen atoms can be 1,6-dihydropyrimidine, 1,2-dihydropyrimidine, 1,4-dihydropyrimidine, 1,6-dihydropyridine, 1,2-dihydropyridine, 1,4-dihydropyridine, 2,3-dihydro-1H-pyrrole, 3,4-dihydro-1H-pyrrole, 2,5-dihydro-1H-pyrroleyl, 4H-[1,3,4]thiadiazinyl, 4,5-dihydrooxazolyl, or 4H-[1,4]thiazinyl rings. Unsaturated heterocycles containing oxygen atoms can be 2H-pyran, 4H-pyran, or 2,3-dihydrofuran. Unsaturated heterocycles containing sulfur atoms can be 2H-thiaran or 4H-thiaran. Heterocycles can be benzofused without limitation, such as dihydroisoquinoline rings.

[0036] The term "aryl" refers to a monocyclic, bicyclic, or tricyclic carbocyclic system containing 6-14 ring atoms, 6-12 ring atoms, or 6-10 ring atoms, wherein at least one ring is aromatic. The aryl group is typically, but not necessarily, linked to the parent molecule via its aromatic ring. Examples of aryl groups may include phenyl, naphthyl, and anthracene. The aryl group may optionally be substituted by one or more substituents described in this invention.

[0037] The term "heteroaryl" preferably refers to a monovalent monocyclic, bicyclic, or tricyclic aromatic ring system having 5, 6, 7, 8, 9, or 10 ring atoms ("pentacyclic to decacyclic heteroaryl"), particularly 5, 6, 9, or 10 ring atoms, and the ring atoms contain at least one (preferably 1-4, more preferably 1, 2, or 3) identical or different heteroatoms, said heteroatoms being, for example, oxygen, nitrogen, or sulfur. Furthermore, in each case, the heteroaryl group may be benzofused. Specifically, the heteroaryl group is selected from thienyl, furanyl, pyrroleyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl, thiadiazolyl, etc., and their benzo[derivatives], such as benzofuranyl, benzothienyl, benzooxazolyl, benzoisooxazolyl, benzoimidazolyl, benzotriazolyl, indazole, indolyl, isindolyl, etc.; or pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, etc., and their benzo[derivatives], such as quinolinyl, quinazolinyl, isoquinolinyl, etc.; or azocinyl, inazinyl, purinyl, etc., and their benzo[derivatives]; or cyclophosphinyl, phthalazinyl, quinazolinyl, quinoxolinyl, naphridinyl, carbazole, acridinyl, etc.

[0038] The term "pharmaceutical acceptable" means that, within the bounds of normal medical judgment, contact with a patient's tissues will not cause undue toxicity, irritation, allergic reactions, etc., and that the benefits and risks are reasonable and that the product is effective for its intended use.

[0039] The compounds of this invention can exist in specific geometric or stereoisomeric forms. This invention contemplates all such compounds, including cis and trans isomers, (-)- and (+)- enantiomers, (R)- and (S)- enantiomers, diastereomers, (D)- isomers, (L)- isomers, and racemic mixtures thereof, as well as other mixtures, such as mixtures enriched with enantiomers or diastereomers, all of which are within the scope of this invention. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All such isomers and mixtures thereof are included within the scope of this invention. In some embodiments, the preferred compounds are those isomers exhibiting superior biological activity. Purified or partially purified isomers and stereoisomers of the compounds of this invention, or racemic mixtures or diastereomer mixtures, are also included within the scope of this invention. Purification and separation of such substances can be achieved using standard techniques known in the art.

[0040] Optically pure enantiomers can be obtained by resolving racemic mixtures using conventional methods, such as by forming diastereomer salts using optically active acids or bases, or by forming covalent diastereomers. Mixtures of diastereomers can be separated into individual diastereomers based on their physical and / or chemical differences using methods known in the art, such as chromatography or fractional crystallization. An optically active enantiomer base or acid is then released from the separated diastereomer salt. Another method for separating racemic enantiomers uses chiral chromatography (e.g., chiral HPLC columns), where the separated chiral isomers may be conventionally derivatized or not derivatized prior to separation, depending on which method allows for more efficient separation of the chiral isomers. Enzymatic methods can also be used to separate derivatized or underivatized chiral isomers.

[0041] Furthermore, the compounds of the present invention can exist in the form of tautomers. The present invention includes all possible tautomers of the compounds of the present invention, as well as single tautomers or any mixtures of said tautomers in any proportion.

[0042] The present invention also includes all pharmaceutically acceptable isotopically labeled compounds that are identical to the compounds of the present invention, except that one or more atoms are replaced by atoms having the same atomic number but with an atomic mass or mass number different from the dominant atomic mass or mass number in nature.

[0043] The scope of this invention also includes metabolites of the compounds of this invention, i.e., substances formed in the body upon administration of the compounds of this invention. Metabolites of the compounds can be identified using techniques known in the art, and their activity can be characterized by experimental methods. Such products can be generated, for example, by oxidation, reduction, hydrolysis, amidation, deamidation, esterification, enzymatic hydrolysis, etc., of the administered compound. Therefore, this invention includes metabolites of the compounds of this invention, including compounds obtained by methods that expose the compounds of this invention to mammals for a time sufficient to produce their metabolites.

[0044] The term "pharmaceutically acceptable carrier" refers to substances that do not cause significant irritation to the organism and do not impair the biological activity and properties of the active compound. "Pharmaceutically acceptable carriers" include, but are not limited to, glidants, sweeteners, diluents, preservatives, dyes / colorants, flavoring agents, surfactants, wetting agents, dispersants, disintegrants, stabilizers, solvents, or emulsifiers.

[0045] The term "neuropsychiatric disorders" refers to the general term for neurological and mental disorders, including neurological and / or mental disorders.

[0046] The term "cognitive impairment" refers to impairment in one or more cognitive functions, including but not limited to memory, language, visuospatial, executive, calculation, and comprehension and judgment, which affects an individual's daily or social abilities.

[0047] The term "free base" refers to an amine compound in a non-salt form. In this invention, it refers to a non-salt-forming compound of the general formula (A) or its cis-trans isomers, tautomers, enantiomers, diastereomers, meso compounds, racemates, or mixtures thereof.

[0048] The term "hydrochloride salt" refers to a hydrochloric acid addition salt formed by the non-salt-forming compound of general formula (A) or its cis-trans isomers, tautomers, enantiomers, diastereomers, meso compounds, racemates, or mixtures thereof with hydrochloric acid.

[0049] The term "separation" refers to the separation of a compound from a solvent to provide a solid, semi-solid, or slurry. This process is typically accomplished by means such as centrifugation, filtration with or without vacuum, filtration under positive pressure, distillation, evaporation, or a combination thereof. Separation may or may not be accomplished by purification as the chemical purity, chiral purity, or both of the chemical purity and chiral purity of the separated product are increased. Purification is typically accomplished by means such as crystallization, distillation, extraction; filtration through acidic, basic, or neutral alumina; filtration through acidic, basic, or neutral activated carbon; column chromatography on a column packed with a chiral stationary phase; filtration through porous paper, plastic, or glass barriers; column chromatography on silica gel; ion exchange chromatography; recrystallization; normal-phase high-performance liquid chromatography; reversed-phase high-performance liquid chromatography; grinding, etc.

[0050] In the following examples, unless the chirality of the compound is specified, it refers to the meso compound, racemic compound, or mixture thereof.

[0051] The hydrochloride salt of the compound of the present invention

[0052] This invention provides a hydrochloride salt in the form of a compound represented by the following general formula (A), or a cis-trans isomer, tautomer, enantiomer, diastereomer, meso compound, racemic mixture, or mixture thereof.

[0053]

[0054] in:

[0055] R1 is selected from alkyl, alkenyl, alkynyl, hydroxyl, amino or unsubstituted by one or more alkyl groups, alkoxy, thioalkyl, cyano, haloalkyl, cycloalkyl, heterocyclic, substituted or unsubstituted heteroaryl, and substituted or unsubstituted aryl.

[0056] R2 and each R3 may be the same or different, and each is independently selected from hydrogen, halogen, alkyl, alkenyl, alkynyl, alkoxy, thioalkyl, haloalkyl, cyano, amino groups substituted with or unsubstituted with one or more alkyl groups, cycloalkyl and heterocyclic groups.

[0057] R4, R5 and each of R6 may be the same or different, and each is independently selected from hydrogen, alkyl, haloalkyl, hydroxy, alkoxy, thioalkyl, halogen and cyano;

[0058] R7 is selected from hydrogen, alkyl, haloalkyl, and...

[0059] n1 is any integer between 0 and 1; n2 is any integer between 0 and 3; n3 is any integer between 1 and 2; n4 is any integer between 1 and 3; n5 is any integer between 1 and 3.

[0060] The alkyl, alkoxy, and thioalkyl groups are each optionally and independently substituted with one or more substituents selected from halogens, C1-C6 alkoxy groups, and Rx groups; the alkyl group in the haloalkyl group is optionally substituted with one or more Rx substituents; and the cycloalkyl and heterocyclic groups are each optionally and independently substituted with one or more substituents selected from halogens, Rx groups, C1-C6 alkyl groups, C1-C6 haloalkyl groups, C1-C6 alkoxy groups, and C1-C6 thioalkyl groups.

[0061] The aryl group is selected from phenyl or naphthyl, wherein the phenyl or naphthyl group is optionally substituted by one or more substituents selected from halogen, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 thioalkyl and Rx;

[0062] The heteroaryl group is a five- to ten-membered heteroaryl group, wherein its ring atom contains one, two or three heteroatoms selected from oxygen, nitrogen and sulfur, preferably containing one or two heteroatoms selected from oxygen, nitrogen and sulfur, and the heteroaryl group is optionally substituted by one or more substituents selected from halogen, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 thioalkyl and Rx;

[0063] Each time Rx appears, it is independently selected from -OH, -NH2, -NO2, and -CN;

[0064] y is 0.5 to 2.5, for example 0.5, 1, 1.5, 2, 2.5 or 3, preferably 0.5, 1, 1.5 or 2.

[0065] In a preferred embodiment of the present invention, the hydrochloride salt of the compound represented by general formula (A) or its cis-trans isomers, tautomers, enantiomers, diastereomers, meso compounds, racemates, or mixtures thereof,

[0066] in:

[0067] R1 is selected from C1-C6 alkyl, hydroxyl, C1-C6 alkoxy, cyano, C1-C6 haloalkyl, C3-C6 cycloalkyl, phenyl, wherein the phenyl is optionally substituted by one or more substituents selected from halogen, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 thioalkyl and Rx;

[0068] R2 and each R3 may be the same or different, and each is independently selected from hydrogen, halogen, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 thioalkyl, C1-C6 haloalkyl, cyano, C1-C6 alkyl-substituted amino, C3-C6 cycloalkyl and 3-8 membered heterocyclic groups;

[0069] R4, R5 and each of R6 may be the same or different, and each is independently selected from hydrogen, C1-C6 alkyl, C1-C6 haloalkyl, hydroxyl, C1-C6 alkoxy, C1-C6 thioalkyl, halogen and cyano;

[0070] R7 is selected from hydrogen, C1-C6 alkyl, C1-C6 haloalkyl, and...

[0071] n1 is any integer between 0 and 1; n2 is any integer between 0 and 3; n3 is any integer between 1 and 2; n4 is any integer between 1 and 3; n5 is any integer between 1 and 3.

[0072] The 3-8 membered heterocyclic group contains 1, 2 or 3 heteroatoms selected from oxygen, nitrogen and sulfur in its ring atoms, preferably containing 1 or 2 heteroatoms selected from oxygen, nitrogen and sulfur in its ring atoms, and the heterocyclic group is optionally substituted by one or more substituents selected from halogen, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 thioalkyl and Rx;

[0073] Each time Rx appears, it is independently selected from -OH, -NH2, -NO2, and -CN;

[0074] y is 0.5 to 2.5, for example 0.5, 1, 1.5, 2, 2.5 or 3, preferably 0.5 to 2, for example 0.5, 1, 1.5 or 2.

[0075] In a preferred embodiment of the present invention, the hydrochloride salt of the compound represented by general formula (A) or its cis-trans isomers, tautomers, enantiomers, diastereomers, meso compounds, racemates, or mixtures thereof,

[0076] in:

[0077] R1 is selected from methyl, ethyl, cyano, hydroxy, phenyl, trifluoromethyl, trichloromethyl, cyclopropyl, cyclobutyl, ethoxy, methoxy, phenyl, trifluoroethyl, trichloroethyl, isopropyl, n-propyl, hydroxyethyl, cyanomethyl, and methoxyethyl;

[0078] R2 and each R3 may be the same or different, and each is independently selected from hydrogen, fluorine, chlorine, bromine, methyl, cyano, thiomethyl, methoxy, Trifluoromethyl, trichloromethyl, cyclopropyl and -N(CH3)2;

[0079] R4, R5 and each R6 may be the same or different, and each is independently selected from hydrogen and methyl;

[0080] R7 is selected from hydrogen, methyl, and...

[0081] n1 is any integer between 0 and 1 independently; n2 is any integer between 0 and 1 independently; n3 is any integer between 1 and 2 independently; n4 is 1 independently; n5 is any integer between 1 and 2 independently.

[0082] y is selected from 0.5, 1, 1.5 or 2, with 1 being preferred.

[0083] In a preferred embodiment of the present invention, the hydrochloride salt of the compound represented by general formula (A) or its cis-trans isomers, tautomers, enantiomers, diastereomers, meso compounds, racemates, or mixtures thereof,

[0084] in:

[0085] R1 is selected from methyl, ethyl, cyano, hydroxy, phenyl, trifluoromethyl, cyclopropyl, cyclobutyl, ethoxy, methoxy, isopropyl, n-propyl, hydroxyethyl, cyanomethyl, and methoxyethyl;

[0086] R2 and each R3 may be the same or different, and each is independently selected from hydrogen, fluorine, chlorine, methyl, cyano, thiomethyl, methoxy, Trifluoromethyl, cyclopropyl and -N(CH3)2;

[0087] R4, R5 and each R6 may be the same or different, and each is independently selected from hydrogen and methyl;

[0088] R7 is selected from hydrogen, methyl, and...

[0089] n1 is any integer between 0 and 1; n2 is independently any integer between 0 and 1; n3 is independently any integer between 1 and 2; n4 is independently 1; n5 is independently any integer between 1 and 2.

[0090] y is selected from 0.5 to 2, for example 0.5, 1, 1.5 or 2, preferably 1.

[0091] In a preferred embodiment of the present invention, the hydrochloride salt of the compound represented by general formula (A) or its cis-trans isomers, tautomers, enantiomers, diastereomers, meso compounds, racemates, or mixtures thereof is characterized in that it comprises hydrochloride salts selected from the following compounds:

[0092]

[0093]

[0094] In a preferred embodiment of the present invention, the hydrochloride salt of the above-mentioned specific compound is preferably a dihydrochloride, and more preferably a monohydrochloride.

[0095] The present invention further provides hydrochloride salts of compounds of the following general formula (A-1) or their cis-trans isomers, tautomers, enantiomers, diastereomers, meso compounds, racemates, or mixtures thereof:

[0096]

[0097] R1 is selected from methyl, ethyl, cyano, hydroxy, phenyl, trifluoromethyl, trichloromethyl, cyclopropyl, cyclobutyl, ethoxy, methoxy, trifluoroethyl, trichloroethyl, isopropyl, n-propyl, hydroxyethyl, cyanomethyl, and methoxyethyl;

[0098] R2 is selected from hydrogen, fluorine, chlorine, bromine, methyl, cyano, thiomethyl, methoxy, Trifluoromethyl, trichloromethyl, cyclopropyl and -N(CH3)2;

[0099] y is selected from 1 or 2, preferably 1.

[0100] In a preferred embodiment of the present invention, the hydrochloride salt of the compound represented by general formula (A-1) or its cis-trans isomers, tautomers, enantiomers, diastereomers, meso- or racemic mixtures thereof, wherein R1 is selected from methyl, ethyl, n-propyl and methoxyethyl, more preferably methyl and ethyl; and R2 is selected from cyano and chlorine, more preferably chlorine.

[0101] In a more preferred embodiment of the present invention, the hydrochloride salt of the compound represented by general formula (A-1) or its cis-trans isomers, tautomers, enantiomers, diastereomers, meso- or racemic mixtures or mixtures thereof, wherein R1 is selected from methyl and ethyl; and R2 is chlorine.

[0102] This invention also provides hydrochloride salts of compound NH200102 or its cis-trans isomers, tautomers, enantiomers, diastereomers, meso compounds, racemates, or mixtures thereof.

[0103]

[0104] y is selected from 1 or 2, preferably 1.

[0105] In one embodiment of the present invention, the hydrochloride salt of compound NH200102 is selected from the hydrochloride salt of compound NH200102-A or the hydrochloride salt of compound NH200102-B as shown below:

[0106]

[0107] y is selected from 1 or 2, preferably 1.

[0108] In a preferred embodiment of the present invention, in the hydrochloride salt of the compound NH200102 or its cis-trans isomer, tautomer, enantiomer, diastereomer, meso compound, racemic compound or mixture thereof, the molar ratio of the free base of the compound NH200102 to the hydrochloric acid is 1:1.

[0109] In a preferred embodiment of the present invention, in the hydrochloride salt of the compound NH200102 or its cis-trans isomer, tautomer, enantiomer, diastereomer, meso compound, racemic compound or mixture thereof, the chemical ratio of the free base of the compound NH200102 to the hydrochloric acid is 1:1.

[0110] In a preferred embodiment of the present invention, the monohydrochloride salt of compound NH200102 is shown below:

[0111]

[0112] In a preferred embodiment of the present invention, the monohydrochloride salt of compound NH200102-A or the monohydrochloride salt of compound NH200102-B are as follows:

[0113]

[0114] In one embodiment of the present invention, the monohydrochloride salt of compound NH200102 is preferably the monohydrochloride salt of compound NH200102-A:

[0115]

[0116] In one embodiment of the present invention, the monohydrochloride salt of compound NH200102 is preferably the monohydrochloride salt of compound NH200102-B:

[0117]

[0118] This invention also provides hydrochloride salts of compound NH200003 or its cis-trans isomers, tautomers, enantiomers, diastereomers, meso compounds, racemates, or mixtures thereof.

[0119]

[0120] y is selected from 1 or 2, preferably 1.

[0121] In one embodiment of the present invention, the hydrochloride salt of compound NH200003 is selected from the hydrochloride salt of compound NH200003-A, or the hydrochloride salt of compound NH200003-B, as shown below:

[0122]

[0123] y is selected from 1 or 2, preferably 1.

[0124] In a preferred embodiment of the present invention, in the hydrochloride salt of the compound NH200003 or its cis-trans isomer, tautomer, enantiomer, diastereomer, meso compound, racemic compound or mixture thereof, the molar ratio of the free base of the compound NH200003 to the hydrochloric acid is 1:1.

[0125] In a preferred embodiment of the present invention, in the hydrochloride salt of the compound NH200003 or its cis-trans isomer, tautomer, enantiomer, diastereomer, meso compound, racemic compound or mixture thereof, the chemical ratio of the free base of the compound NH200003 to the hydrochloric acid is 1:1.

[0126] In a preferred embodiment of the present invention, the monohydrochloride salt of compound NH200003 is shown below:

[0127]

[0128] In a preferred embodiment of the present invention, the monohydrochloride salt of compound NH200003-A or the monohydrochloride salt of compound NH200003-B are as follows:

[0129]

[0130] In one embodiment of the present invention, the monohydrochloride salt of compound NH200003 is preferably the monohydrochloride salt of compound NH200003-A:

[0131]

[0132] In one embodiment of the present invention, the monohydrochloride salt of compound NH200003 is preferably the monohydrochloride salt of compound NH200003-B:

[0133]

[0134] In one embodiment of the present invention, the hydrochloride salt of the compound represented by general formula (A) or its cis-trans isomers, tautomers, enantiomers, diastereomers, meso compounds, racemates, or mixtures thereof comprises its solvate form.

[0135] Preparation method

[0136] This invention provides a method for preparing a hydrochloride salt of a compound represented by general formula (A) or its cis-trans isomers, tautomers, enantiomers, diastereomers, meso compounds, racemates, or mixtures thereof, comprising:

[0137] A solution is prepared by dissolving the free base of the compound of general formula (A) or its cis-trans isomers, tautomers, enantiomers, diastereomers, meso compounds, racemates, or mixtures thereof in an organic solvent. Hydrochloric acid is optionally dissolved in the organic solvent. Then, the hydrochloric acid or hydrochloric acid solution is added to the free base solution of the compound of general formula (A), and the mixture is stirred at room temperature for 0.1 to 24 hours. The mixture is then separated and dried to obtain the hydrochloride salt of the compound of general formula (A) or its cis-trans isomers, tautomers, enantiomers, diastereomers, meso compounds, racemates, or mixtures thereof. The organic solvent includes, but is not limited to, ethyl acetate, acetonitrile, and dichloromethane.

[0138] Monohydrochloride crystal form of compound NH200102

[0139] The present invention also provides a monohydrochloride salt of the compound NH200102-A in crystalline form, defined as crystal form II, with the following structural formula:

[0140]

[0141] In one embodiment of the invention, crystal form II of compound NH200102-A monohydrochloride is provided, and a powder X-ray diffraction pattern expressed in 2θ angles using Cu-Kα radiation is provided: the powder X-ray diffraction pattern of crystal form II includes peaks located at diffraction angles (2θ) of 5.13°±0.2°, 10.26°±0.2°, 12.52±0.2°, 15.42°±0.2°, 17.04°±0.2°, 20.61°±0.2°, 24.98°±0.2°, 25.85°±0.2°, 26.35°±0.2°, 26.86°±0.2° and 32.62°±0.2°.

[0142] Furthermore, the powder X-ray diffraction pattern of crystal form II of compound NH200102-A monohydrochloride includes peaks at diffraction angles (2θ) of 5.13°±0.2°, 10.26°±0.2°, 12.52±0.2°, 15.42°±0.2°, 17.04°±0.2°, 20.61°±0.2°, 21.53°±0.2°, 22.28°±0.2°, 24.98°±0.2°, 25.85°±0.2°, 26.35°±0.2°, 26.86°±0.2°, 29.72°±0.2°, and 32.62°±0.2°.

[0143] More preferably, the powder X-ray diffraction data of crystal form II of compound NH200102-A monohydrochloride are shown in Table 1.1:

[0144] Table 1.1:

[0145]

[0146]

[0147] In a more preferred embodiment of the invention, the crystal form II has essentially the following characteristics: Figure 1 The powder X-ray diffraction pattern shown.

[0148] The crystal form II of the compound NH200102-A monohydrochloride described in this invention can also be characterized by DSC and identified by differential scanning calorimetry at a scanning speed of 10℃ / min, including endothermic peaks (peak temperatures) at 74.8±3℃ and 150.64℃±3℃.

[0149] Pharmaceutical compositions, formulations and kits

[0150] The present invention further provides a pharmaceutical composition comprising a therapeutically effective amount of the compound of general formula (A) or a hydrochloride salt in the form of its cis-trans isomer, tautomer, enantiomer, diastereomer, meso compound, racemic mixture or mixture thereof, and a pharmaceutically acceptable carrier; or a pharmaceutical composition comprising the compound of the present invention, NH200102-A monohydrochloride crystal form II, and a pharmaceutically acceptable carrier.

[0151] In one embodiment of the present invention, the pharmaceutical composition can be formulated using one or more pharmaceutically acceptable carriers in a conventional manner. Therefore, the active compounds of the present invention can be formulated into dosage forms for oral, sublingual, intranasal, parenteral (e.g., intravenous, intramuscular, or subcutaneous), or rectal administration, or suitable for inhalation or insufflation. The hydrochloride salts of the compounds represented by general formula (A) of the present invention, or their cis-trans isomers, tautomers, enantiomers, diastereomers, meso compounds, racemates, or mixtures thereof, can also be formulated into sustained-release dosage forms.

[0152] According to some embodiments of the present invention, the compounds of the present invention can be encapsulated in gelatin capsules or compressed into tablets. For oral therapeutic purposes, the compounds of the present invention can be used with excipients and administered in the form of tablets, lozenges, capsules, suspensions, syrups, etc. According to embodiments of the present invention, the above-described formulations should contain at least 0.5% (w / w) of the active compound of the present invention, but this can vary depending on the specific dosage form, wherein 4% to about 70% by weight is convenient. In such pharmaceutical compositions, the amount of the active compound should reach an appropriate dosage.

[0153] In one embodiment of the invention, regarding oral administration, the active compound of the invention can be formulated into tablets or capsules, for example, by conventional means with pharmaceutically acceptable excipients, such as binders, fillers, lubricants, disintegrants, or wetting agents. Tablets can be coated using methods well known in the art. Liquid formulations for oral administration can be solutions, syrups, or suspensions, or evaporated into a dried product, regenerated with water or other suitable carriers before use. Such liquid formulations can be prepared using pharmaceutically acceptable additives by conventional means, such as suspending agents, emulsifiers, non-aqueous carriers, and preservatives.

[0154] In one embodiment of the present invention, when the active compound of the present invention is used for parenteral administration, the compound provided by the present invention can be combined with sterile water or an organic medium to form an injectable solution or suspension.

[0155] In one embodiment of the present invention, the active compound of the present invention can be formulated into a rectal composition, such as a suppository or retention enema, for example containing a conventional suppository base, such as cocoa butter or other glycerides.

[0156] Treatment methods and uses

[0157] The present invention also provides the use of a compound of general formula (A) or a hydrochloride thereof, in the form of its cis-trans isomer, tautomer, enantiomer, diastereomer, meso compound, racemic mixture, or mixture thereof, in the preparation of a medicament for regulating NMDA receptors. The medicament optionally comprises one or more other active agents that modulate the mammalian nervous system or alleviate mental disorders.

[0158] The present invention also provides a method for modulating NMDA receptors, comprising administering to an individual in need a compound of general formula (A) or a hydrochloride thereof in the form of a cis-trans isomer, tautomer, enantiomer, diastereomer, meso compound, racemic mixture, or mixture thereof, or a pharmaceutical composition thereof. The method may optionally further comprise administering one or more other active agents that modulate the mammalian nervous system or alleviate mental disorders.

[0159] The present invention also provides a hydrochloride salt or pharmaceutical composition thereof in the form of a compound of general formula (A) or a cis-trans isomer, tautomer, enantiomer, diastereomer, meso compound, racemic mixture, or mixture thereof for modulating NMDA receptors. The application optionally further comprises administration of one or more other active agents that modulate the mammalian nervous system or alleviate mental disorders.

[0160] In a preferred embodiment of the present invention, the regulation includes inhibitory activity, inverse agonistic activity, and antagonistic activity of the receptor.

[0161] In a preferred embodiment of the present invention, the hydrochloride salt of the compound represented by general formula (A) or its cis-trans isomers, tautomers, enantiomers, diastereomers, meso- or racemic mixtures thereof acts on the TMD site of the NMDA receptor.

[0162] In another aspect, the present invention also provides the use of the compound of general formula (A) of the present invention, or its cis-trans isomers, tautomers, enantiomers, diastereomers, meso- or racemic isomers, or mixtures thereof, in the form of hydrochloride salts or pharmaceutical compositions of the present invention in the preparation of medicaments for treating neuropsychiatric disorders.

[0163] The present invention also provides a method for treating neuropsychiatric disorders, comprising administering to an individual in need (e.g., a mammalian individual, such as a human) a hydrochloride salt or a pharmaceutical composition thereof in the form of a compound of general formula (A) or a cis-trans isomer, tautomer, enantiomer, diastereomer, meso compound, racemic mixture or mixture thereof.

[0164] The present invention also provides hydrochloride salts or pharmaceutical compositions thereof in the form of compounds of general formula (A) or cis-trans isomers, tautomers, enantiomers, diastereomers, meso- or racemic mixtures thereof, for the treatment of neuropsychiatric disorders.

[0165] In one embodiment, the neuropsychiatric disorders are selected from one or more of pain, schizophrenia, depression, anxiety, sleep disorders, neurodegenerative diseases, cognitive impairment, bipolar disorder, post-traumatic stress disorder, addictive disorders, withdrawal syndrome, or attention deficit, preferably any one or more of pain, depression, anxiety, schizophrenia, sleep disorders, neurodegenerative diseases, cognitive impairment, or bipolar disorder, and more preferably depression, neurodegenerative diseases, cognitive impairment, or pain.

[0166] In a preferred embodiment of the present invention, the pain is selected from acute pain and chronic pain, and the chronic pain includes, but is not limited to, headache, maxillofacial pain, nape of the neck and occipital region pain, neck and shoulder pain, upper limb pain, chest pain, abdominal pain, lower back and leg pain, reproductive tract pain, urinary tract pain, and dysmenorrhea.

[0167] In a further preferred embodiment of the present invention, the pain includes, but is not limited to, traumatic pain, inflammatory pain, ischemic pain, pain caused by metabolic diseases, neurogenic pain, pain caused by tissue or organ malformations, childbirth pain, and pain caused by malignant proliferative diseases.

[0168] In a further preferred embodiment of the present invention, the neurogenic pain includes, but is not limited to, phantom limb pain, stump pain, burning neuralgia, postherpetic neuralgia, sympathetic nerve-related pain, and peripheral nerve pain.

[0169] In a further preferred embodiment of the present invention, the pain caused by the metabolic disease includes, but is not limited to, pain caused by gout and pain caused by diabetes.

[0170] In a further preferred embodiment of the present invention, the pain caused by the malignant proliferative disease includes, but is not limited to, pain caused by tumors, such as leukemia, lymphoma, myeloma, breast cancer, lung cancer, esophageal cancer, stomach cancer, colorectal cancer, liver cancer, pancreatic cancer, head and neck cancer, kidney cancer, bladder cancer, prostate cancer, ovarian cancer, endometrial cancer, cervical cancer, osteosarcoma, soft tissue sarcoma, melanoma, and brain tumors.

[0171] In a further preferred embodiment of the present invention, the pain is selected from moderate to severe pain, and preferably, the moderate to severe pain includes, but is not limited to, traumatic pain, childbirth pain, tumor-induced pain, and inflammatory pain.

[0172] In a preferred embodiment of the present invention, the neurodegenerative disease includes, but is not limited to, Alzheimer's disease, Parkinson's disease, Huntington's disease, and Lewy body dementia, with Alzheimer's disease being more preferred.

[0173] In a preferred embodiment of the present invention, the depression includes, but is not limited to, mild to severe depression, which generally meets the relevant diagnostic criteria of WHOICD-10 (International Classification of Diseases), DSM-5 (Diagnostic and Statistical Manual of Mental Disorders), and CCMD-3 (Chinese Classification and Diagnostic Criteria of Mental Disorders), and is preferably moderate to severe depression.

[0174] In a preferred embodiment of the present invention, the depression includes, but is not limited to, mild depressive episodes, moderate depressive episodes, severe depressive episodes with or without psychotic symptoms, and recurrent depressive episodes.

[0175] Beneficial technical effects of the present invention

[0176] The compound provided by this invention is a reversible NMDAR antagonist with moderate affinity for NMDAR. It specifically targets TMD sites, inhibiting channel opening caused by excessive NMDAR activation under pathological conditions, thereby preventing Ca2+ channel opening. 2+ Excessive influx. Compared with commercially available NMDAR antagonists, the compound provided by this invention can rapidly dissociate after binding to NMDAR, thus not affecting the normal function of NMDAR.

[0177] In vitro studies showed that the compound provided by this invention has essentially the same activity as compound 1a1, but the compound provided by this invention has a faster dissociation rate from NMDAR, which is significantly better than compound 1a1 and comparable to the dissociation rate of NMDAR from methimazole. In addition, rat prepulse inhibition assays showed that the compound provided by this invention is significantly better than compound 1a1 in terms of psychotropic side effects, suggesting that the compound provided by this invention not only has significant analgesic and antidepressant activities, but also has a more prominent therapeutic advantage in reducing psychotropic adverse reactions, and has significant clinical value.

[0178] The present invention also found that, compared with its hydrochloride form, the hydrochloride form of the compound of the present invention significantly increases the exposure of the free base, regardless of whether it is administered via PO or IV route; the bioavailability is significantly improved in the PO route; in addition, the hydrochloride form of the compound of the present invention can significantly improve the solubility compared with the free base, suggesting that the hydrochloride form of the compound of the present invention can reduce the clinical dosage of the free base when used as an API, and has good clinical application prospects and industrial application value. Attached Figure Description

[0179] Figure 1The XRPD spectrum of crystal form II of compound NH200102-A hydrochloride prepared in Examples 3-3 is shown.

[0180] Example

[0181] The embodiments of the present invention are described in detail below. The embodiments described below are exemplary and are only used to explain the present invention, and should not be construed as limiting the present invention. Unless otherwise specified, all proportions, percentages, etc., referred to herein are by weight.

[0182] Differential scanning calorimetry (DSC) images were acquired on either a TA DSC25 or a Mettler Toledo TGA / DSC 3+ simultaneous thermal analyzer. The method parameters were as follows: heating rate: 10 °C / min; protective gas: nitrogen.

[0183] The powder X-ray diffraction pattern was acquired using a Panaco Aeris X-ray powder diffractometer at a standard temperature, such as 25°C. The X-ray powder diffraction method parameters are as follows: X-ray reflection parameters: Cu, Kα; wavelength: Tube voltage: 40KV; Tube current: 15mA; Step size: 0.0110°; Scan time per step: 18.87s; Scan range: from 3.0 to 40.0 degrees.

[0184] Synthesis Examples

[0185] Example 1-1: Preparation of the compound represented by general formula (A)

[0186] The preparation was carried out in accordance with the methods disclosed in patent applications PCT / CN2020 / 129826 or CN114269747A.

[0187] Example 1-1: Preparation of the hydrochloride salt of the compound shown in general formula (A)

[0188] The compound of general formula (A) is dissolved in an organic solvent to form a solution. Hydrochloric acid is optionally dissolved in an organic solvent. Then, hydrochloric acid or hydrochloric acid solution is added to the solution of the compound of general formula (A). The mixture is stirred at room temperature for 0.1 to 24 hours. The solution is then separated and dried to obtain the hydrochloride salt of the compound of general formula (A). The organic solvent includes, but is not limited to, ethyl acetate, acetonitrile, and dichloromethane.

[0189] Example 2-1: Preparation of 3-amino-6′-chloro-2′-ethyl-1′,2′-dihydro-3′-hydro-spiro[cyclobutane-1,4′-isoquinoline]-3′-one (compound NH200003)

[0190]

[0191] Step 1: 2-(3-chlorophenyl)-N-ethylacetamide (NH200003-int 1)

[0192] In a 100 mL single-necked flask, 5 g of 3-chlorophenylacetic acid (29.31 mmol) was dissolved in 50 mL of dichloromethane. Then, 4.53 g of thionyl chloride (38.10 mmol) and 1.07 g of N,N-dimethylformamide (14.66 mmol) were added at 0 °C, and the mixture was stirred at room temperature for 1 hour under nitrogen protection. The solvent was then concentrated using an oil pump to remove excess solvent, and the solution was dissolved in 25 mL of dichloromethane. 29.3 mL of ethylamine (2 M THF) (58.62 mmol) was added at 0 °C, and the mixture was stirred at room temperature for 12 hours. After the reaction was complete, 50 mL of water and 40 mL of dichloromethane were added for extraction to obtain the organic phase. The organic phase was concentrated under reduced pressure to obtain the crude product, which was separated by column chromatography (dichloromethane:methanol = 30:1, v / v) to give 3.26 g of the title compound as a yellowish-brown solid, with a yield of 56.3%.

[0193] Step 2: 6-Chloro-2-ethyl-1,4-dihydroisoquinoline-3(2H)-one (NH200003-int 2)

[0194] In a 50 mL single-necked flask, 2.26 g of NH₂₀₀₀₃-int 1 (11.43 mmol), 412 mg of paraformaldehyde (13.72 mmol), and 15 mL of Eaton's reagent were added. The mixture was stirred at 80 °C for 2 hours under nitrogen protection. After the reaction was complete, 70 mL of water and 25 mL of dichloromethane were added for extraction to obtain the organic phase. The organic phase was concentrated under reduced pressure to obtain the crude product, which was separated by column chromatography (petroleum ether: ethyl acetate = 5:1, v / v) to give 1.7 g of yellow solid A, with a yield of 70.8%.

[0195] Step 3: tert-butyl((1,3-dibromoprop-2-yl)oxy)dimethylsilane (NH200003-int 3)

[0196] In a 250 mL single-necked flask, 10 g of 1,3-dibromo-2-propanol (45.89 mmol), 6.25 g of imidazole (91.78 mmol), and 100 mL of dichloromethane were added. 8.3 g of tert-butyldimethylchlorosilane (TBSCl, 55.07 mmol) was added at 0 °C, and the mixture was stirred at room temperature for 12 hours. After the reaction was complete, 200 mL of water and 100 mL of dichloromethane were added, and the mixture was extracted to obtain the organic phase. The organic phase was concentrated under reduced pressure to obtain the crude product, which was separated by column chromatography (petroleum ether:ethyl acetate = 30:1, v / v) to give 14.96 g of the title compound as a colorless liquid, with a yield of 99.7%.

[0197] Step 4: 3-((tert-butyldimethylsilyl)oxy)-6′-chloro-2′-ethyl-1′,2′-dihydro-3′H-spiro[cyclobutane-1,4′-isoquinoline]-3′-one (NH200003-int 4)

[0198] In a 100 mL single-necked flask, 1 g of NH₂₀₀₀₃-int 2 (4.77 mmol) was dissolved in 20 mL of tetrahydrofuran (THF), followed by 2 mL of hexamethylphosphoric triamine (HMPA). Under nitrogen protection, 4.2 mL of n-butyllithium (10.49 mmol) was added dropwise at -40 °C, and the mixture was stirred for 1 hour at this temperature. Then, 1.9 g of NH₂₀₀₀₃-int 3 (5.72 mmol) in 5 mL of THF solution was added, and the reaction was stirred at room temperature for 12 hours. After the reaction was complete, 40 mL of water and 25 mL of ethyl acetate were added to extract the organic phase. The organic phase was concentrated under reduced pressure to obtain the crude product, which was separated by column chromatography (petroleum ether:ethyl acetate = 5:1, v / v) to give 760 mg of a yellow oil, with a yield of 42.0%.

[0199] Step 5: 6′-Chloro-2′-Ethyl-3-hydroxy-1′,2′-dihydro-3′H-spiro[cyclobutane-1,4′-isoquinoline]-3′-one (NH200003-int 5)

[0200] In a 50 mL single-necked flask, 760 mg of NH₂₀₀₀₃-int 4 (2.0 mmol) was dissolved in 10 mL of tetrahydrofuran (THF), and 3 mL of tetrabutylammonium fluoride (TBAF, 3.0 mmol) was added. The mixture was stirred at room temperature for 1 hour. After the reaction was complete, 40 mL of water and 25 mL of ethyl acetate were added for extraction to obtain the organic phase. The organic phase was concentrated under reduced pressure to obtain the crude product, which was separated by column chromatography (petroleum ether:ethyl acetate = 1:1, v / v) to give 460 mg of a yellow oil, with a yield of 86.6%.

[0201] Step 6: 2-(6′-chloro-2′-ethyl-3′-oxo-2′,3′-dihydro-1′H-spiro[cyclobutane-1,4′-isoquinoline]-3-yl)isoindoline-1,3-dione (NH200003-int 6)

[0202] In a 50 mL single-necked flask, 460 mg of NH₂₀₀₀₃-int 5 (1.73 mmol), 305 mg of phthalimide (2.08 mmol), and 545 mg of triphenylphosphine (PPh₃, 2.08 mmol) dissolved in 5 mL of tetrahydrofuran (THF) were added. 362 mg of diethyl azodicarbonate (DEAD, 2.08 mmol) was added at 0 °C, and the mixture was stirred for 12 hours under nitrogen protection, gradually decreasing to room temperature. After the reaction was complete, 40 mL of water and 25 mL of ethyl acetate were added for extraction to obtain the organic phase. The organic phase was concentrated under reduced pressure to obtain the crude product, which was then separated using a pre-prepared plate (petroleum ether:ethyl acetate = 1:1, v / v) to yield 340 mg of crude product.

[0203] Step 7: 3-Amino-6′-chloro-2′-ethyl-1′,2′-dihydro-3′-hydro-spiro[cyclobutane-1,4′-isoquinoline]-3′-one (NH200003)

[0204] In a 50 mL single-necked flask, 340 mg of the product from the previous step (0.86 mmol) and 5 mL of ethanolamine were added, and the mixture was stirred at 70 °C for 1 hour. After the reaction was complete, 20 mL of water and 15 mL of dichloromethane were added for extraction to obtain the organic phase. The organic phase was concentrated under reduced pressure to obtain the crude product, which was separated by pre-prepared plate separation (dichloromethane:methanol = 10:1, v / v) to obtain 30 mg of a colorless oil, with a yield of 13.2% and a purity of 97.3%.

[0205] 1 H NMR (400MHz, CDCl3) δ7.44(s,1H),7.19(d,J=8.0Hz,1H),7.08(d,J=8.0Hz,1H),4.33(s,2H),3.8 5–3.81(m,1H),3.59-3.54(m,2H),3.07-3.02(m,2H),2.08-2.03(m,2H),1.18-1.15(m,3H);[M+H] + 265.1

[0206] Example 2-2: trans-3-amino-6′-chloro-2′-ethyl-1′,2′-dihydro-3′H-spiro[cyclobutane-1,4′-isoquinoline]-3′-one (compound NH200003-A) and cis-3-amino-6′-chloro-2′-ethyl-1′,2′-dihydro-3′H-spiro[cyclobutane-1,4′-isoquinoline]-3′-one (compound NH200003-B)

[0207] NH200003 was prepared by HPLC (Mobile Phase A: H2O (0.1% TFA), Mobile Phase B: CAN, Column: C18, 5µm, 4.6x250mm, Flow rate: 15mL·min). -1 NH200003-A and NH200003-B can be obtained by separation.

[0208] NH200003-A: 1 H NMR(400MHz,Methanol-d4)δ7.61(s,1H),7.31–7.25(m,2H),4.49(s,2H),3.97-3.88(m,1 H),3.61-3.56(m,2H),3.02-2.96(m,2H),2.45-2.38(m,2H),1.20(t,J=7.2Hz,3H);[M+H] + 265.1

[0209] NH200003-B: 1 H NMR(400MHz, CDCl3)δ7.66(d,J=2.0Hz,1H),7.35–7.28(m,2H),4.53(s,2H),4.12–3.97(m, 1H),3.64–3.56(m,2H),3.09–2.97(m,2H),2.60–2.45(m,2H),1.21(t,J=7.2Hz,3H);[M+H] + 265.1

[0210] Examples 2-3: Preparation of NH200003 or NH200003-A or NH200003-B monohydrochloride

[0211]

[0212] Weigh out crude NH200003 or NH200003-A or NH200003-B (10.6 mg, 0.04 mmol), dissolve in 0.5 mL acetonitrile, add 4 μL concentrated hydrochloric acid (37%), stir at room temperature for 20 hours, centrifuge, and dry the resulting solid under vacuum for 4 hours.

[0213] 1) Cl was determined by ion chromatography. - The actual content is 11.65%, and when it forms one molecule of hydrochloride, Cl - The theoretical content is 11.64%, and the stoichiometric ratio of NH200003 or NH200003-A or NH200003-B to hydrochloric acid is 1:1.

[0214] 2)1 H NMR (400MHz, DMSO-d6): δ8.23 (s, 3H), δ7.73 (d, J = 4.0Hz, 1H), δ7.36-7.30 (m, 2H), δ4.48 (s, 2H), δ3. 92-3.81(m,1H), δ3.50-3.45(m,2H), δ2.84-2.79(m,2H), 2.55-2.51(m,2H), δ1.09(t,J=8.0Hz,3H).

[0215] 3) Analysis and identification by differential scanning calorimetry showed that the scanning speed was 10℃ / min, including an endothermic peak at 95.19℃ (peak temperature) and a melting endothermic peak at 259.80℃ (peak temperature), with an error tolerance of ±3℃.

[0216] Example 3-1: Preparation of 3-amino-6′-chloro-2′-methyl-1′,2′-dihydro-3′H-spiro[cyclobutane-1,4′-isoquinoline]-3′-one (compound NH200102)

[0217] The target compound was prepared by replacing ethylamine with methylamine according to the method in Example 2-1, with a purity of 99.40%.

[0218] 1 H NMR (400MHz, CDCl3) δ7.45 (s, 1H), 7.20 (d, J = 8.4Hz, 1H), 7.06 (d, J = 8.4Hz, 1H), 4.37 (s,2H),3.89-3.81(m,1H),3.11(s,3H),3.08-3.02(m,2H),2.06-2.00(m,2H);[M+H] + 251.0

[0219] Examples 3-2: trans-3-amino-6′-chloro-2′-methyl-1′,2′-dihydro-3′H-spiro[cyclobutane-1,4′-isoquinoline]-3′-one (compound NH200102-A) and cis-3-amino-6′-chloro-2′-methyl-1′,2′-dihydro-3′H-spiro[cyclobutane-1,4′-isoquinoline]-3′-one (compound NH200102-B)

[0220] NH200102 was prepared by HPLC (Mobile Phase A: H2O (0.1% TFA), Mobile Phase B: CAN, Column: C18, 5µm, 4.6x250mm, Flow rate: 15mL·min). -1 The compounds NH200102-A and NH200102-B can be obtained by separation.

[0221] NH200102-A: 1 H NMR (400MHz, Methanol-d4) δ7.58 (d, J = 4.0Hz, 1H), 7.26-7.20 (m, 2H), 4.47 (s, 2 H),3.75-3.67(m,1H),3.10(s,3H),2.98-2.92(m,2H),2.20-2.13(m,2H);[M+H] + 251.1

[0222] NH200102-B: 1 H NMR(400MHz,Methanol-d4)δ7.67(d,J=4.0Hz,1H),7.32-7.24(m,2H),4.53(s,2 H),4.12-4.03(m,1H),3.12(s,3H),3.05-3.02(m,2H),2.53-2.27(m,2H).;[M+H] + 251.1

[0223] Example 3-3: Preparation of NH200102 or NH200102-A or NH200102-B hydrochloride

[0224]

[0225] Crude NH200102 or NH200102-A or NH200102-B (3.34 g) was dissolved in 20 mL of dichloromethane, and 20 mL of 2 mol / L ethyl acetate solution of hydrogen chloride was added dropwise. The mixture was stirred at room temperature for 0.5 hours, and the solvent was removed by concentration to obtain 4.3 g of pale yellow to white solid.

[0226] 1) Cl was determined by ion chromatography. - The actual content is 11.62%, and when it forms one molecule of hydrochloride, Cl - The theoretical content is 11.64%, and the stoichiometric ratio of NH200102 or NH200102-A or NH200102-B to hydrochloric acid is 1:1.

[0227] 2) 1 H NMR (400MHz, DMSO-d6): δ8.41(s,3H),7.83(s,1H),7.34(d,J=8.2Hz,1H),7.26(d,J=8.2H z,1H),4.49(s,2H),3.94-3.88(m,1H),3.01(s,3H),2.84-2.79(m,2H),2.58-2.52(m,2H).

[0228] 3) Analysis using differential scanning calorimetry showed that the scan rate was 10℃ / minute, and the calorimeter contained endothermic peaks at 74.8℃ and 150.64℃, with an error tolerance of ±3℃.

[0229] 4) The separated NH₂₀₁₀₂-A was identified by Aeris X-ray powder diffraction (XRPD) using Cu-Kα radiation. It exhibited the following characteristic peaks expressed in 2θ angles: 5.13°, 10.26°, 12.52°, 15.42°, 17.04°, 20.61°, 24.98°, 25.85°, 26.35°, 26.86°, 31.13°, and 32.62°, with an error tolerance of ±0.2°, and was defined as crystal form II. The XRPD data for crystal form II are shown in Table 1.1 above, and the XRPD plot is shown below. Figure 1 As shown.

[0230] Test case

[0231] Test Example 1: NMDAR Inhibitory Activity

[0232] Test drug: Compound 1a1 was prepared according to the method in CN106957285A, and its structure is shown below:

[0233]

[0234] NH200102 and NH200003 were prepared according to the above examples.

[0235] Negative control: Weigh an appropriate volume of DMSO as the storage solution.

[0236] Test compound: Weigh an appropriate mass of the compound (actual amount = theoretical concentration × volume × molecular weight / purity). Calculate the required volume of DMSO using the formula, and then convert this to the final required mass of DMSO. Next, dissolve the powder in the weighed DMSO using ultrasonication. Calculate the actual stock solution concentration based on the final amount of DMSO used. Generally, the actual stock solution concentration will slightly differ from the theoretical concentration. If there are solubility issues, adjust the stock solution concentration accordingly.

[0237] Preparation method of working solution for drug delivery formulation: Before channel current testing, dilute the negative control and test compound stock solution to an appropriate amount of extracellular fluid to prepare the working solution. See the table below for details:

[0238] Table 1. Administration methods and test concentrations

[0239]

[0240] Experimental System

[0241] Cell line selection

[0242] In this experiment, the HEK293 cell line stably expressing the rat NR1 / NR2B genes was used, with gene numbers NR1: NM_017010; NR2B: NM_012574.

[0243] Concentration selection criteria

[0244] In this experiment, the detection concentrations of the test compounds were 0.3 μM, 1 μM, 3 μM, 10 μM, and 30 μM. The negative control was 0.1% DMSO.

[0245] Preparation of the liquid for testing

[0246] Extracellular fluid: 140mM NaCl, 4mM KCl, 2mM CaCl2, 10mM HEPES, 5mM D-Glucose pH=7.4 (NaOH).

[0247] Intracellular fluid: 10mM NaCl, 135mM CsMes, 2mM MgCl2, 10mM HEPES, 10mM EGTA, 2mM Na2-ATP, 0.2mM Na2-GTP, pH=7.2 (CsOH).

[0248] After preparing the electrode internal solution, aliquot it into 1 mL tubes and store them at -20°C. Use freshly thawed electrode internal solution for each experiment. All electrode internal solutions should be used within one month. After one month, discard the old electrode internal solution and prepare fresh solution.

[0249] The patch clamp testing method is as follows:

[0250] The capillary glass tube (BF150-86-10, Sutter Instruments) was drawn into a recording electrode using a microelectrode drawing instrument (P97, Sutter Instruments). Under an inverted microscope (Olympus, IX71), the microelectrode manipulator (Sutter Instruments, MP285) was used to bring the recording electrode into contact with the cell, and negative pressure was applied to aspirate and form a GΩ seal. After forming the GΩ seal, rapid capacitance compensation was performed, and then negative pressure was continued to rupture the cell membrane, forming a whole-cell recording mode. Slow capacitance compensation was then performed, and the membrane capacitance and series resistance were recorded. No leakage compensation was applied.

[0251] A coverslip containing cells was placed in the recording bath of an inverted microscope to form a whole-cell seal. Under normal extracellular fluid perfusion, NMDA and the test compound were sprayed onto the cell surface using a rapid jet drug delivery system, and the NMDA current was recorded. The test compound reaching the cell surface was rapidly carried away by the extracellular fluid. Multiple cells were independently and repeatedly tested. All electrophysiological experiments were performed at room temperature.

[0252] The voltage stimulation protocol for whole-cell patch-clamp recording of whole-cell NMDA currents is as follows: After whole-cell sealing is achieved, the cell membrane voltage is clamped at -70mV or -60mV. The drug administration sequence is as follows:

[0253] Step 1: Extracellular fluid (NMDA 100μM + glycine 10μM);

[0254] Step 2: Extracellular fluid (NMDA 100μM + glycine 10μM + test compound (0.3μM));

[0255] Step 3: Extracellular fluid (NMDA 100μM + glycine 10μM);

[0256] Step 4: Extracellular fluid (NMDA 100μM + glycine 10μM + test compound (1μM));

[0257] Step 5: Extracellular fluid (NMDA 100μM + glycine 10μM);

[0258] Step 6: Extracellular fluid (NMDA 100μM + glycine 10μM + test compound (3μM));

[0259] Step 7: Extracellular fluid (NMDA 100μM + glycine 10μM);

[0260] Step 8: Extracellular fluid (NMDA 100μM + glycine 10μM + test compound (10μM));

[0261] Step 9: Extracellular fluid (NMDA 100μM + glycine 10μM);

[0262] Step 10: Extracellular fluid (NMDA 100μM + glycine 10μM + test compound (30μM)).

[0263] Data Analysis

[0264] First, the current and blank control current after treatment with different concentrations of each analyte were standardized, and the corresponding inhibition rates were calculated. For each concentration, the mean and standard error were calculated, and the half-maximal inhibitory concentration (WMC) of each analyte was calculated using the following equation:

[0265] inhibition = 1 / [1+(IC) 50 / C) h ]

[0266] The dose-dependent effect was nonlinearly fitted using the above equation, where C represents the drug concentration, and IC50 is the concentration of the drug. 50 The half-inhibitory concentration (IC50) is represented by h, which represents the Hill coefficient. Curve fitting and IC50 are also mentioned. 50 The calculations were performed using the IGOR software.

[0267] Table 2 Results of NMDAR inhibitory activity test

[0268] Test compounds IC50 (μM) Compound 1a1 0.99 NH200102 1.1 NH200003 3.7

[0269] in conclusion

[0270] The above results show that the compounds provided by the present invention have good inhibitory activity against NMDAR, and the activities of NH200102 or NH200003 are basically equivalent to or similar to those of compound 1a1, suggesting the potential for good clinical therapeutic effects.

[0271] Test Example 2: Dissociation Rate Detection

[0272] Test drugs: Compounds 1a1, NH200102, and NH200003; ketamine and memantine were purchased from Merck Life Sciences (Shanghai) Co., Ltd.

[0273] Positive controls: N-methyl-D-aspartic acid (NMDA), purchased from Merck Life Sciences (Shanghai) Co., Ltd.; Glycine, purchased from Merck Life Sciences (Shanghai) Co., Ltd.

[0274] Preparation method of drug delivery preparation stock solution

[0275] Test reagent: Weigh an appropriate amount of the test sample. Calculate the required volume of DMSO using the formula: DMSO volume = actual amount × purity / (molecular weight × theoretical concentration). Take the corresponding volume of DMSO. Dissolve the weighed test sample in the taken-up DMSO, and simultaneously weigh the DMSO. Calculate the actual stock solution concentration based on the final amount of DMSO used. Generally, the actual stock solution concentration will slightly differ from the theoretical concentration. If there are solubility issues, adjust the stock solution concentration accordingly.

[0276] Positive control: Weigh appropriate amounts of NMDA and Glycine and prepare a 100 mM stock solution with ultrapure water. Aliquot and store at -20°C.

[0277] Preparation method of working solution for drug delivery formulation

[0278] Before the NMDA / NR2B current test, the positive control stock solution and the test sample stock solution were taken out from -20°C and diluted into an appropriate amount of extracellular fluid as the working solution.

[0279] The test concentration of the sample is diluted with DMSO to prepare a diluent, which is then diluted with extracellular fluid to the working solution concentration. The concentration of DMSO in each working solution is 0.1%. For detailed preparation instructions, please refer to the Aisiyipu Compound Working Solution Preparation Record Sheet. The test sample working solution should be used before patch-clamp testing.

[0280] The test sample stock solution and positive control stock solution are stored at -20℃. The test sample working solution and positive control working solution are prepared on the day of testing and stored at room temperature.

[0281] Cell culture

[0282] HEK cell line stably expressing the rNR2B channel, gene information: rNR1:Grin1,NM_017010;

[0283] rNR2B:Grin2b,NM_012574

[0284] Cells were cultured in DMEM medium containing 10% fetal bovine serum and 10 μg / mL blasticidin, 100 μg / mL zeocin, and 200 μg / mL hygromycin B at 37°C and 5% carbon dioxide.

[0285] Cell passage: Remove the old culture medium and wash once with PBS, then add 1 mL of 0.25% Trypsin-EDTA solution and incubate at 37°C for 1 min. When the cells detach from the bottom of the dish, add 5 mL of preheated (37°C) complete culture medium. Gently pipette the cell suspension to separate aggregated cells. Transfer the cell suspension to sterile centrifuge tubes and centrifuge at 1000 rpm for 5 min to collect the cells. For expansion or maintenance culture, seed cells into 6 cm cell culture dishes at a rate of 2.5 × 10⁵ cells per dish (final volume: 5 mL).

[0286] To maintain the electrophysiological activity of cells, the cell density must not exceed 80%.

[0287] For patch-clamp assays, cells were separated using 0.25% Trypsin-EDTA before the experiment. 2 × 10⁴ cells were seeded onto coverslips and tetracycline and DAP5 were added, then cultured in 24-well plates (final volume: 500 μL). The assays were performed 18 hours later.

[0288] Patch clamp testing

[0289] Record the liquid used

[0290] Extracellular fluid: 140mM NaCl, 4mM KCl, 2mM CaCl2·2H2O, 10mM HEPES (4-hydroxyethylpiperazine ethanesulfonic acid), 5mM D-Glucose, pH adjusted to 7.4 with NaOH.

[0291] Intracellular fluid: 110mM cesium methanesulfonate, 10mM NaCl, 2mM MgCl2·6H2O, 10mM EGTA (ethylene glycol bis(2-aminoethyl ether)tetraacetic acid), 2mM Na2-ATP, 0.2mM Na2-GTP, 10mM HEPES, pH adjusted to 7.2 with CsOH.

[0292] After preparation, intracellular fluid was aliquoted into 1 mL tubes and stored at -20°C. Freshly thawed intracellular fluid was used daily for experiments. All intracellular fluid was used within one month. After one month, the old intracellular fluid was discarded and freshly prepared.

[0293] Patch clamp testing

[0294] A capillary glass tube was drawn into a recording electrode using a microelectrode drawing instrument. Under an inverted microscope, the microelectrode manipulator was used to bring the recording electrode into contact with the cell, and negative pressure was applied to aspirate and form a GΩ seal. After forming the GΩ seal, rapid capacitance compensation (pF) was performed, and then negative pressure was continued to rupture the cell membrane, forming a whole-cell recording mode. Slow capacitance compensation was then performed, and the membrane capacitance (pF) and series resistance were recorded. No leakage compensation was applied.

[0295] A coverslip containing cells was placed in the recording bath of an inverted microscope. The working solution of the test sample and the NMDA+Glycine external solution without the compound were perfused through the recording bath by gravity to act on the cells. Liquid exchange was carried out using a vacuum pump during recording. Multiple data were collected for each concentration. All electrophysiological experiments were performed at room temperature.

[0296] The voltage stimulation protocol for whole-cell patch-clamp recording of NR2B currents is as follows: After whole-cell sealing, the cell membrane voltage is clamped at -70 mV. Recording is performed in gap-free mode for 600 seconds. The specific drug administration method is as follows: first, NMDA + Glycine is administered until the current stabilizes; then, a mixture of the test sample, NMDA, and Glycine is administered until the current stabilizes; finally, the sample is rinsed with NMDA + Glycine for 3-5 minutes. Experimental data were acquired using an EPC-10 amplifier (HEKA) and stored in PatchMaster (HEKA) software.

[0297] Data Analysis

[0298] The current used for flushing with NMDA+Glycine after each drug treatment was nonlinearly fitted, and the time constant (Tau, τ) when the current recovers to a steady state was calculated using the following equation:

[0299]

[0300] The Halflife value was obtained by dividing the Tau value by 1.44269504. Curve fitting was performed using IGOR software. The results are shown in Table 3 below:

[0301] Table 3 Results of dissociation rate inhibition activity test

[0302] compound Tau(s) Halflife(s) Megatron 4.13±0.24 2.86±0.16 NH200102 4.28±0.55 2.97±0.38 Compound 1a1 27.32±5.27 18.94±3.65 ketamine 16.80±3.93 11.65±2.73

[0303] in conclusion

[0304] The above dissociation rate test results show that the dissociation rate of the compound NH200102 of the present invention is significantly faster than that of other tested drug compounds 1a1 and ketamine, and comparable to that of memantine, suggesting that NH200102 of the present invention can rapidly dissociate from NMDAR and has the potential for good clinical treatment tolerability.

[0305] Test Example 3: Pharmacokinetic Test

[0306] The free base and monohydrochloride samples of NH200102-A were dissolved in physiological saline (injection grade) to prepare the test sample.

[0307] This study used LC-MS to investigate the pharmacokinetic characteristics of NH200102-A free base and hydrochloride in rats under two administration routes (IV: intravenous injection; PO: oral administration).

[0308] Experimental Design:

[0309] Animal drug administration

[0310] Four SD rats in each group were fasted for 12 hours before the experiment, but had free access to water. They were administered the above-mentioned free base and monohydrochloride solutions according to the prescribed dosage and administration method (iv: 1 mg / kg; PO: 10 mg / kg; calculated as free base). In the iv group, 0.3 mL of blood was collected from the orbital sinus at 2 min, 5 min, 15 min, 30 min, 1 h, 2 h, 4 h, 6 h, and 8 h after administration. In the PO group, 0.3 mL of blood was collected at 5 min, 15 min, 30 min, 1 h, 2 h, 4 h, 6 h, 8 h, and 10 h after administration. The plasma was separated by centrifugation and stored at -20℃ for later analysis. The pharmacokinetic parameters of the free base and monohydrochloride in rats are shown in Table 4.

[0311] Table 4. Pharmacokinetic parameters of free base and monohydrochloride in rats

[0312]

[0313] in conclusion

[0314] Compared with the monohydrochloride, the free base of NH200102-A showed significantly higher exposure levels with the monohydrochloride, regardless of whether it was administered via PO or IV route. The bioavailability was also significantly improved with the PO route. This suggests that the hydrochloride of the compound of this invention can reduce the clinical dosage of the free base when used as an API, and has good prospects for clinical application.

[0315] Test Example 4: Solubility Test

[0316] Test sample: Free base and monohydrochloride sample of NH200102-A

[0317] 1. Prepare a pH 6.8 phosphate buffer solution medium according to the 2020 Chinese Pharmacopoeia, Part IV, General Chapter 8004;

[0318] 2. Using the above buffer solution as a solvent, prepare a supersaturated solution of the sample to be tested;

[0319] 3. After a brief vortexing, incubate the supersaturated solution at 37°C for 5 minutes. Then continue vortexing at 25°C for 24 hours.

[0320] 4. Pour 200 μL of the incubation solution into a filter plate (0.4 μm, Millipore) and filter to obtain the filtrate;

[0321] 5. Prepare a standard curve (10-5000 ng / mL);

[0322] 6. Dilute the supernatant solution to be tested to a concentration detectable by HPLC-UV;

[0323] 7. The prepared standard curve solution and the supernatant to be tested were added sequentially to the HPLC-UV for detection. Two parallel groups were set up for each sample. The solubility of the free base of NH200102 was measured to be 39.4 mg / mL, and the solubility of NH200102 monohydrochloride was 140.59 mg / mL (equivalent to 123.3 mg / mL of free base). The above results show that the solubility of NH200102 monohydrochloride is significantly better than that of the free base of NH200102.

[0324] Patent applications PCT / CN2020 / 129826 or CN114269747A describe the structure of the free base of the compound of this invention, its preparation method, and the detection method and results of its NMDAR inhibitory activity. The entire contents of PCT / CN2020 / 129826 or CN114269747A are incorporated herein by reference. The inventors have further confirmed through experiments that the hydrochloride salt of the compound provided by this invention has the same or similar NMDAR inhibitory activity as the free base.

[0325] Those skilled in the art will recognize that many modifications and variations can be made to the invention without departing from its spirit and scope. The specific embodiments described herein are provided by way of example only and are not intended to limit the scope in any way. The true scope and spirit of the invention are shown in the appended claims, and the description and embodiments are merely exemplary.

Claims

1. A hydrochloride salt of a compound represented by the following general formula (A-1) or its cis-trans isomer: ， in: R1 is selected from methyl or ethyl; R2 is selected from chlorine; y is selected from 1 or 2.

2. The hydrochloride salt of the compound of general formula (A-1) as claimed in claim 1, characterized in that, The hydrochloride salts of the following compound NH200102 are selected: ， y is selected from 1 or 2.

3. The compound of claim 2, or a salt of its cis-trans isomer with hydrochloric acid, wherein y is 1.

4. The compound of claim 2, or a salt of its cis-trans isomer with hydrochloric acid, wherein Selected from the hydrochloride salts of compound NH200102-A or compound NH200102-B as shown below: ， ， y is selected from 1 or 2.

5. The hydrochloride salt of the compound of general formula (A-1) as described in claim 4, characterized in that, y is 1.

6. The hydrochloride salt of the compound of general formula (A-1) as claimed in claim 1, characterized in that, The hydrochloride salts of the following compound NH200003 are selected: ， y is selected from 1 or 2.

7. The hydrochloride salt of the compound of general formula (A-1) as described in claim 6, characterized in that, y is 1.

8. The hydrochloride salt of the compound of general formula (A-1) as described in claim 6, characterized in that, The hydrochloride salt of compound NH200003 is selected from the hydrochloride salt of compound NH200003-A or the hydrochloride salt of compound NH200003-B as shown below: or , y is selected from 1 or 2.

9. The hydrochloride salt of the compound of general formula (A-1) as claimed in claim 8, characterized in that, y is 1.

10. The monohydrochloride salt of the compound NH 200102-A in crystalline form, defined as Form II, characterized by, The structural formula of the monohydrochloride salt is shown below. Using Cu-Kα radiation, the powder X-ray diffraction pattern expressed in 2θ angles shows that the powder X-ray diffraction pattern of crystal form II includes peaks at diffraction angles (2θ) of 5.13°±0.2°, 10.26°±0.2°, 12.52±0.2°, 15.42°±0.2°, 17.04°±0.2°, 20.61°±0.2°, 24.98°±0.2°, 25.85°±0.2°, 26.35°±0.2°, 26.86°±0.2° and 32.62°±0.2°.

11. The monohydrochloride salt of compound NH200102-A in crystalline form as described in claim 10, characterized in that, The powder X-ray diffraction pattern of crystal form II includes peaks at diffraction angles (2θ) of 5.13°±0.2°, 10.26°±0.2°, 12.52±0.2°, 15.42°±0.2°, 17.04°±0.2°, 20.61°±0.2°, 21.53°±0.2°, 22.28°±0.2°, 24.98°±0.2°, 25.85°±0.2°, 26.35°±0.2°, 26.86°±0.2°, 29.72°±0.2°, and 32.62°±0.2°.

12. The monohydrochloride salt of compound NH200102-A in crystalline form according to claim 10, characterized in that, The crystal form II has one or more of the following characteristics: (1) It has a basic XRPD diffraction pattern as shown in Figure 1; (2) Characterized by DSC and analyzed by differential scanning calorimetry at a scanning speed of 10℃ / min, it has endothermic peaks with peak temperatures of 74.8±3℃ and 150.64℃±3℃.

13. A pharmaceutical composition, characterized by, The compound comprising a therapeutically effective amount of the compound of general formula (A-1) as claimed in any one of claims 1-9 or its cis-trans isomer hydrochloride, or the compound NH200102-A monohydrochloride crystal form II as claimed in any one of claims 10-12 and a pharmaceutically acceptable carrier.

14. The use of the hydrochloride salt of the compound of general formula (A-1) as described in any one of claims 1-9 or the cis-trans isomer thereof, or the monohydrochloride form II of the compound NH200102-A as described in any one of claims 10-12, or the use of the pharmaceutical composition as described in claim 13 in the preparation of a medicament for treating neuropsychiatric diseases.

15. The use as described in claim 14, characterized in that, The neuropsychiatric disorders mentioned are selected from one or more of the following: pain, schizophrenia, depression, anxiety, sleep disorders, neurodegenerative diseases, cognitive impairment, bipolar disorder, post-traumatic stress disorder, addictive disorders, withdrawal syndrome, or attention deficit.

16. The use as described in claim 14, characterized in that, The neuropsychiatric disorders mentioned are selected from any one or more of the following: pain, depression, anxiety, schizophrenia, sleep disorders, neurodegenerative diseases, cognitive impairment, or bipolar disorder.

17. The use of claim 14, wherein, The neuropsychiatric disorders mentioned are depression, neurodegenerative diseases, cognitive impairment, or pain.

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