Salts of 1,5-dihydro-2,4-benzodiazepin-3-one derivatives and their applications

Abstract

The present invention belongs to the field of medicine, and particularly relates to salts of compounds represented by the following general formula I, preparation methods, compositions containing the salts of the compounds, and applications in the field of medicine. #imgabs0#

Description

Technical Field

[0001] The present invention belongs to the field of medicine, and particularly relates to a salt of a 1,5-dihydro-2,4-benzodiazepin-3-one derivative, a preparation method, a composition comprising the salt of the compound, and an application in the field of medicine. -3-one derivative salts, preparation methods, and compositions containing such compound salts, as well as their use in the medical field. Technical Background

[0002] Schizophrenia has a hidden onset and a low admission rate, with a relatively high lifetime prevalence. Currently, approximately 0.3-0.7% of the world's population is affected by schizophrenia during their lifetime, and it is estimated that there are more than 21 million schizophrenia patients globally in 2016. Currently, the main antipsychotic drugs for schizophrenia are typical antipsychotic drugs and atypical antipsychotic drugs. However, current schizophrenia treatment drugs strongly block dopamine receptors, resulting in adverse reactions such as extrapyramidal reactions (EPS), tardive dyskinesia, and increased prolactin. In the medical field, although there are various types of active compounds acting on different targets available for the treatment of sleep disorders, adverse reactions such as addiction, drug resistance, and residual effects remain unsolved problems.

[0003] Traditionally, antipsychotic drugs that exert pharmacological effects by blocking dopamine D2 receptors are called first-generation antipsychotic drugs, that is, "typical" antipsychotic drugs (such as haloperidol). They have breakthroughs in treating the positive symptoms of schizophrenia but fail to treat negative symptoms and cognitive impairments. Typical antipsychotic drugs generally have severe EPS side effects and are ineffective in one-third of schizophrenia patients.

[0004] After the 1960s, a series of new-generation antipsychotic drugs were successively developed, including ziprasidone, risperidone, etc., which are called second-generation antipsychotic drugs, that is, new antipsychotic drugs. Although their respective pharmacological effects are not completely the same, they have common pharmacological characteristics, that is, their affinities for 5-hydroxytryptamine (5-HT) receptors (5-HT1A, 2A, 2c) and norepinephrine (NA) receptors (α1, α2) are much higher than those for D2 receptors, resulting in a relatively high D2 / 5-HT 2A ratio. Their clinical effects have more advantages compared with first-generation antipsychotic drugs. They are not only as effective as traditional antipsychotic drugs in treating positive symptoms but also effective in treating negative symptoms and cognitive deficit symptoms, with a wider spectrum of action. However, these drugs have adverse reactions such as QT interval prolongation, hyperprolactinemia, and weight gain. Therefore, finding drugs that are effective in treating the positive and negative symptoms and cognitive impairments of schizophrenia and have fewer side effects is a current research hotspot.

[0005] The serotonin system plays an important role in regulating the functions of the prefrontal cortex (PFC), including emotional control, cognitive behavior, and working memory. Pyramidal neurons and GABA interneurons in the PFC contain several serotonin receptor subtypes 5-HT 1A and 5-HT 2A . Recently, it has been demonstrated that the PFC and NMDA receptor channels are targets of 5-HT 1A R, and these two receptors regulate excitatory neurons in the cerebral cortex, thereby affecting cognitive function. In fact, various preclinical data suggest that 5-HT 1A R may be a new target for the development of antipsychotic drugs. The high affinity of atypical antipsychotics (such as olanzapine, aripiprazole, etc.) for 5-HT 1A R and their low EPS side effects all indicate that the serotonin system plays an important role in regulating the functions of the prefrontal cortex (PFC), including emotional control, cognitive behavior, and working memory. Pyramidal neurons and GABA interneurons in the PFC contain several serotonin receptor subtypes 5-HT 1A and 5-HT 2A . Recent studies have shown that 5-HT 1A agonists are related to the treatment of atypical antipsychotics and can improve negative symptoms and cognitive impairment. In the treatment of schizophrenia with the atypical antipsychotic clozapine, it has been found that 5-HT 2A plays a very important role, involving all aspects of perception, emotional regulation, and motor control. Blocking the 5-HT 2A receptor can normalize the release of dopamine and play an antipsychotic role. In addition, the 5-HT 2C receptor is closely related to weight gain.

[0006] Pimavanserin is an inverse agonist with high affinity for 5-HT 2A and 5-HT 2C . The in vitro experimental results show that its affinity for the 5-HT 2A receptor [inhibition constant (Ki) is 0.4 nM] is higher than that for 5-HT 2C (Ki = 16 nM), and it has no obvious affinity (Ki > 300 nM) for 5-HT 2B receptors, dopamine receptors (including D2 receptors), adrenergic receptors, muscarinic receptors, or calcium channel receptors. This drug was approved by the US Food and Drug Administration for marketing in April 2016 under the trade name NuplazidTM and is mainly used for the treatment of Parkinsonian psychosis symptoms such as hallucinations and illusions.

[0007] Therefore, there is a need to find an antipsychotic drug that is effective against both positive and negative symptoms, can improve cognitive impairment, can prevent extrapyramidal side effects, including tardive dyskinesia and Parkinson's disease, and can reduce weight gain. Summary of the Invention

[0008] Patent application PCT / CN2021 / 089660 protects a series of 1,5-dihydro-2,4-benzodiazepin -3-one derivatives. In the subsequent research and development process, in order to maximize the therapeutic effect of the above active substances in clinical research and to simplify product handling and improve product solubility in the subsequent pharmaceutical research and development process, the present invention has conducted a comprehensive and systematic study on the salts of the above substances in order to obtain the most suitable salt form.

[0009] All the contents involved in patent application PCT / CN2021 / 089660 are incorporated into the present invention by citation.

[0010] The present invention aims to provide salts of 1,5-dihydro-2,4-benzodiazepin -3-one derivatives. Compared with the free base form of the compound, phosphates or citrates or crystal forms are beneficial to improving the solubility and hygroscopic properties of the active substance, enhancing the bioavailability and stability of the active substance, and facilitating subsequent clinical development and production development.

[0011] The present invention provides a salt of a compound represented by the following general formula I. Preferably, the salt form of the compound represented by general formula I is a phosphate or a citrate.

[0012]

[0013] The present invention also provides the use of the phosphate or citrate of the compound represented by the above general formula I in the preparation of a drug for treating neuropsychiatric diseases.

[0014] In vitro research results show that the compound provided by the present invention has basically the same activity as pimavanserin, but the compound provided by the present invention has lower 5-HT 2A 、5-HT 2C receptor Ki values, which are significantly better than those of pimavanserin, indicating that the compound provided by the present invention is more likely to bind to the receptor and has great clinical application value.

[0015] The present invention is implemented by the following technical solutions:

[0016] On the one hand, the present invention provides a salt of a compound represented by the following general formula I.

[0017]

[0018] Wherein:

[0019] n1 and n2 are integers from 1 to 3;

[0020] R1 is selected from linear or branched C1-C8 alkyl, C2-C8 alkenyl and C2-C8 alkynyl, and the alkyl, alkenyl and alkynyl are each independently and optionally substituted with substituents selected from halogen and C1-C8 haloalkyl;

[0021] R2 is selected from hydrogen, halogen and C1-C8 haloalkyl;

[0022] R3, R4, R5, R6 are each independently selected from hydrogen, halogen and C1-C8 haloalkyl;

[0023] R7 is selected from linear or branched C1-C8 alkyl, cycloalkyl and R8 and R9 are each independently selected from linear or branched C1-C8 alkyl, and the alkyl and cycloalkyl are optionally substituted with substituents selected from halogen and C1-C8 haloalkyl;

[0024] Z is selected from C, O, N;

[0025] Q and W are each independently selected from C, N.

[0026] In a preferred embodiment of the present invention, the salt form of the compound represented by the general formula I is phosphate or citrate, as shown in Formula II and Formula III:

[0027]

[0028] y is 0.5, 1, 1.5, 2, 2.5 or 3, preferably 0.5, 1, 1.5 or 2.

[0029] In a preferred embodiment of the present invention, the phosphate or citrate of the compound represented by the general formula I, wherein:

[0030] The linear or branched C1-C8 alkyl is selected from linear or branched C1-C5 alkyl and linear or branched C1-C3 alkyl; and / or

[0031] The C2-C8 alkenyl is C2-C5 alkenyl; and / or

[0032] The C2-C8 alkynyl is C2-C5 alkynyl; and / or

[0033] The C1-C8 haloalkyl is C1-C5 haloalkyl; and / or

[0034] The cycloalkyl is C3-C 10 cycloalkyl, preferably C3-C6 cycloalkyl.

[0035] In a preferred embodiment of the present invention, the phosphate or citrate of the compound represented by the general formula I, wherein:

[0036] The halogen mentioned above is selected from fluorine, chlorine, bromine, iodine; and / or

[0037] The straight-chain or branched C1-C5 alkyl group is selected from methyl, ethyl, propyl, isopropyl, butyl, isobutyl, pentyl, isopentyl; and / or

[0038] The C3-C6 cycloalkyl group is selected from cyclopropyl, cyclobutyl, cyclopentyl; and / or

[0039] The straight-chain or branched C1-C3 alkyl group is selected from methyl, ethyl, propyl, isopropyl. In a preferred embodiment of the present invention, the phosphate or citrate of the compound represented by the general formula I, wherein:

[0040] The halogen mentioned above is fluorine, chlorine; and / or

[0041] The straight-chain or branched C1-C5 alkyl group is selected from methyl, ethyl, propyl, isopropyl, isobutyl; and / or

[0042] The C3-C6 cycloalkyl group is selected from cyclopropyl, cyclobutyl; and / or

[0043] The straight-chain or branched C1-C3 alkyl group is selected from methyl, ethyl, propyl, isopropyl;

[0044] The is

[0045] In a preferred embodiment of the present invention, the phosphate or citrate of the compound represented by the general formula I is characterized in that it includes the phosphate or citrate of the compound selected from the following:

[0046]

[0047]

[0048] In one embodiment of the present invention, the phosphate or citrate of the compound represented by the general formula I includes its solvate form.

[0049] In one embodiment of the present invention, there is provided an A19006 citrate as shown in formula IV:

[0050]

[0051] In one embodiment of the present invention, there is provided an A19006 phosphate as shown in formula V:

[0052]

[0053] On the other hand, the present invention provides polymorphic form I of A19006 citrate represented by formula IV, which is characterized in that, using Cu-Kα radiation, an X-ray powder diffraction pattern expressed in terms of diffraction angle 2θ±0.2° is obtained, which shows characteristic peaks at 10.03, 12.62, 13.69, 14.68, 16.44, 17.19, 19.10, 20.27, 21.77, wherein the error range of 2θ for each characteristic peak is ±0.2.

[0054] In a preferred embodiment of the present invention, the X-ray powder diffraction pattern of the polymorphic form I shows characteristic peaks expressed in terms of 2θ±0.2° at 17.67, 19.57, 21.30, 23.78, 24.03, 25.57, 26.22, wherein the error range of 2θ for each characteristic peak is ±0.2.

[0055] In a preferred embodiment of the present invention, the X-ray powder diffraction pattern of the polymorphic form I shows characteristic peaks expressed in terms of 2θ±0.2° at 5.42, 10.03, 11.10, 12.62, 13.69, 14.68, 16.44, 17.19, 17.67, 18.39, 19.10, 19.57, 20.27, 21.30, 21.77, 22.50, 23.78, 24.03, 25.57, 26.22, 27.65, 28.62, 30.29, 30.56, 31.67, 32.76, 33.62, 34.04, 34.88, 35.49, 36.33, 39.02, 39.69, wherein the error range of 2θ for each characteristic peak is ±0.2.

[0056] In a more preferred embodiment of the present invention, the melting endothermic peak value of the DSC of the polymorphic form I is selected from 164.7–170.0 °C, preferably 167.7 °C.

[0057] Preparation method

[0058] The present invention provides a method for preparing a salt of a compound represented by general formula I, comprising:

[0059] Dissolving the free base of the compound represented by general formula I in an organic solvent to form a solution, optionally dissolving an acid in an organic solvent, dropping the organic solvent-acid solution into the free base solution of the compound represented by general formula I, stirring, and separating and drying to obtain the salt of the compound represented by general formula I; the organic solvents include but are not limited to absolute ethanol, ethyl acetate, etc.; the acids include but are not limited to phosphoric acid, citric acid, etc. Detailed implementation mode

[0060] 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 belongs. In case of conflict, the definitions provided in this application shall prevail. When trade names appear herein, they are intended to refer to their corresponding goods or their active ingredients. All patents, published patent applications and publications cited herein are incorporated herein by reference.

[0061] General terms and definitions

[0062] The term "comprising" is an open-ended expression, i.e., it includes the content specified in the present invention, but does not exclude other aspects. It should be understood that the term "comprising" can cover a closed meaning, i.e., "consisting of".

[0063] As described in the present invention, the compounds of the present invention may optionally be substituted by one or more substituents, such as the compounds of the general formula above or specific examples and subclasses as in the examples. It should be understood that the term "optionally substituted" and the term "substituted or unsubstituted" can be used interchangeably. Generally, the term "substituted" means that one or more hydrogen atoms in the given structure are replaced by specific substituents, provided that the normal valence of the specified atoms in the current situation is not exceeded and the substitution forms a stable compound. Combinations of substituents and / or variables are permitted only when such combinations form stable compounds. When it is described that a certain substituent is absent, it should be understood that the substituent can be one or more hydrogen atoms, provided that the structure enables the compound to reach a stable state. Unless otherwise indicated, an optionally substituted group can be substituted at each substitutable position of the group. When more than one position in the given structural formula can be substituted by one or more substituents selected from a specific group, then the substituents can be the same or different at each position.

[0064] Unless otherwise specified, as used herein, the point of attachment of a substituent can be from any suitable position of the substituent. When the bond of a substituent is shown as passing through the bond connecting two atoms in a ring, then such a substituent can be bonded to any ring-forming atom in the substitutable ring.

[0065] In addition, it should be noted that, unless otherwise explicitly indicated, the description method "are each independently" adopted in the present invention should be understood in a broad sense, which can either mean that among different groups, the specific options expressed between the same symbols do not affect each other, or mean that within the same group, the specific options expressed between the same symbols do not affect each other.

[0066] When the lower and upper limits of a numerical range are disclosed, any numerical value and any included range falling within that range are specifically disclosed. In particular, each range of values disclosed herein should be understood to represent every numerical value and range subsumed within the broader range. When any variable (e.g., R), and variables with subscripts (e.g., R1, R2, R3, R4, R5, R6, R7, etc.) occur more than once in the composition or structure of a compound, their definitions are independent of each other in each occurrence. For example, if a group is substituted with 0, 1, 2, 3, or 4 R substituents, the group may optionally be substituted with up to four R substituents, and the options for each R substituent in each case are independent of each other.

[0067] In various parts of this specification, the substituents of the compounds disclosed in the present invention are disclosed according to group types or ranges. In particular, the present invention includes each independent secondary combination of the individual members of these group types and ranges. For example, the expression m - n used herein refers to the range from m to n, as well as the sub - ranges and individual point values composed of the point values therein. For example, the term "C1 - C5 alkyl" specifically refers to methyl, ethyl, C3 alkyl, C4 alkyl, C5 alkyl independently disclosed. For example, the expression "C2 - C8" or "C 2-8 " encompasses the range of 2 - 8 carbon atoms and should be understood to also encompass any sub - ranges and each point value therein, such as C2 - C5, C3 - C4, C2 - C6, C3 - C6, C4 - C6, C4 - C7, C4 - C8, C2 - C4, etc., and C2, C3, C4, C5, C6, C7, C8, etc. Again, for example, the expression "C1 - C5" or "C 1-5 " encompasses the range of 1 - 5 carbon atoms and should be understood to also encompass any sub - ranges and each point value therein, such as C2 - C5, C3 - C4, C1 - C2, C1 - C3, C1 - C4, C1 - C5, etc., and C1, C2, C3, C4, C5, etc. Again, for example, the expression "C2 - C5" or "C 2-5 " encompasses the range of 2 - 5 carbon atoms and should be understood to also encompass any sub - ranges and each point value therein, such as C2 - C5, C3 - C4, C2 - C3, C2 - C4, C3 - C5, C4 - C5, etc., and C2, C3, C4, C5, etc. Again, for example, the expression "C1 - C8" or "C 1-8”covers the range of 1 to 8 carbon atoms and should be understood to also cover any sub-ranges therein, as well as each point value, such as C2-C5, C3-C4, C2-C6, C3-C6, C4-C6, C4-C7, C4-C8, C2-C4, etc., and C1, C2, C3, C4, C5, C6, C7, C8, etc. For another example, the expression "ternary to octavalent" should be understood to cover any sub-ranges and each point value therein, such as ternary to pentavalent, ternary to hexavalent, ternary to heptavalent, ternary to octavalent, quaternary to pentavalent, quaternary to hexavalent, quaternary to heptavalent, quaternary to octavalent, pentavalent to heptavalent, pentavalent to octavalent, hexavalent to heptavalent, hexavalent to octavalent, etc., and three, four, five, six, seven, eight valent, etc. Other similar expressions in this text should be understood in a similar manner.

[0068] The ranges listed herein (such as numerical ranges) can cover each value within the range and each sub-range formed by the respective values. Thus, for example, the expression "n2 is any integer between 0 and 3" includes, for example, any integer from 0 to 2, any integer from 2 to 3, etc., such as 1, 2, 3.

[0069] The term "one or more" or a similar expression "at least one" can mean, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more.

[0070] The term "selected from..." means one or more elements independently selected from the group listed hereinafter, and can include combinations of two or more elements.

[0071] When it is described that each carbon atom in a group can optionally be replaced by a heteroatom, provided that the normal valence of all atoms in the group in the current case is not exceeded and a stable compound is formed.

[0072] The term "hydrogen (H)" represents a single hydrogen atom, and such an atomic group can be connected to other groups, for example, connected to an oxygen atom to form a hydroxyl group.

[0073] The term "halogen" or "halo" should be understood to mean fluorine (F), chlorine (Cl), bromine (Br) or iodine (I), preferably fluorine, chlorine, bromine atoms, more preferably fluorine atom, chlorine atom.

[0074] The term "alkyl" refers to a straight-chain or branched-chain saturated aliphatic hydrocarbon group composed of carbon atoms and hydrogen atoms, which is connected to the rest of the molecule through a single bond. "Alkyl" can have 1 to 6 carbon atoms, that is, "C1-C6 alkyl", for example, C 1-4 alkyl, C 1-3 alkyl, C 1-2 alkyl, C3 alkyl, C4 alkyl, C 1-6 alkyl, C3-6 An alkyl group. It can also have 1 - 3 carbon atoms, i.e., "C1 - C3 alkyl", such as C 1-3 alkyl, C 1-2 alkyl, C3 alkyl. Again, for example, the term "C1 - C5 alkyl" specifically refers to methyl, ethyl, C3 alkyl, C4 alkyl, and C5 alkyl independently disclosed. 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 (-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.

[0075] The term "alkenyl" refers to a straight - chain or branched - chain unsaturated aliphatic hydrocarbon group composed of carbon atoms and hydrogen atoms and having at least one double bond. The alkenyl group can have 2 - 5 carbon atoms, i.e., "C 2-5 alkenyl", such as C 2-4 alkenyl, C 3-4Alkenyl. 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, and the like.

[0076] The term "cycloalkyl" refers to a saturated cyclic hydrocarbon group composed of carbon atoms and hydrogen atoms, preferably containing 1 or 2 rings. The cycloalkyl can be a monocyclic, fused polycyclic, bridged or spiro ring structure. The cycloalkyl can have 3 - 6 carbon atoms, i.e., "C3-C6 cycloalkyl", such as C6 cycloalkyl, C5 cycloalkyl, C4 cycloalkyl, C3 cycloalkyl. Non-limiting examples of cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, etc. This term also covers the case where the C atoms can be substituted by oxo (=O).

[0077] The term "alkynyl" refers to a straight-chain or branched-chain unsaturated aliphatic hydrocarbon group having at least one triple bond composed of carbon atoms and hydrogen atoms. The alkynyl can have 2 - 5 carbon atoms, i.e., "C 2-5 alkynyl", such as C 2-3 alkynyl, C 2-4 alkynyl. Non-limiting examples of alkynyl include, but are not limited to, ethynyl, prop-1-ynyl, prop-2-ynyl, but-1-ynyl, but-2-ynyl, but-3-ynyl, etc.

[0078] The term "pharmaceutically acceptable carrier" refers to those substances that have no obvious irritating effect on 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.

[0079] The term "free base" refers to an amine compound in non-salt form, and in the present invention, it refers to the compound represented by general formula I in non-salted form.

[0080] The "salt" herein refers to a pharmaceutically acceptable salt formed by an acid and a base, specifically hydrochloride, phosphate, citrate, tartrate, maleate, fumarate, acetate, succinate, malate, mandelate, oxalate, dihydrogen phosphate, hydrogen phosphate, bisulfate, sulfate, formate, chloroacetate, glycolate, trifluoroacetate, propionate, acrylate, butyrate, isobutyrate, valerate, pivalate, hexanoate, benzoate, phenylacetate, oxalate, malonate, succinate, maleate, fumarate, glutarate, adipate, phthalate, isophthalate, terephthalate, etc.

[0081] The "y" refers to the chemical ratio of the base and a pharmaceutically acceptable acid, and y can be 0.5, 1, 1.5, 2, 2.5 or 3, preferably 0.5, 1, 1.5 or 2.

[0082] The "crystal form" refers to the ordered arrangement of molecules with the same chemical structure in a certain manner. In this application, it specifically refers to Crystal Form I of A19006 citrate.

[0083] The "X-ray powder diffraction pattern" or "XRPD" as described in the present invention refers to according to Bragg's formula 2d sinθ = nλ (where λ is the wavelength of the X-ray, the diffraction order n is any positive integer, generally taking the first-order diffraction peak, n = 1), when the X-ray is incident on an atomic plane with a d lattice plane spacing of a crystal or a partial crystal sample at a grazing angle θ (the complementary angle of the incident angle, also known as the Bragg angle), the Bragg equation can be satisfied, and thus this set of X-ray powder diffraction patterns can be measured.

[0084] The "2θ" or "2θ angle" as described in the present invention refers to the diffraction angle, θ is the Bragg angle, with the unit of ° or degree, and the error range of 2θ is from ±0.1 to ±0.5, preferably from ±0.1 to ±0.3, more preferably ±0.2.

[0085] The "interplanar spacing" or "interplanar spacing (d value)" as described in the present invention refers to that in the space lattice, three non-parallel unit vectors a, b, c connecting adjacent two lattice points are selected, and they divide the lattice into juxtaposed parallelepiped units, which are called interplanar spacings. The space lattice is divided by connecting lines according to the determined parallelepiped unit to obtain a set of straight grids, which are called space lattices or crystal lattices. The lattice and the crystal lattice respectively reflect the periodicity of the crystal structure with geometric points and lines. For different crystal planes, their interplanar spacings (i.e., the distance between adjacent two parallel crystal planes) are different; the unit is or Å.

[0086] The "differential scanning calorimetry" or "DSC" as described in the present invention measures the transition temperature when the crystal absorbs or releases heat due to the change of its crystal structure or crystal melting. For the same crystal form of the same compound, in continuous analysis, the error of the thermal transition temperature and the melting point can be within about 5 °C, usually within about 3 °C. When describing that a certain compound has a given DSC peak or melting point, it refers to the DSC peak or melting point ±5 °C. "Basically" also takes into account such temperature changes. DSC provides an auxiliary method for distinguishing different crystal forms. Different crystal forms can be identified according to their different transition temperature characteristics. It should be noted that for mixtures, their DSC peaks or melting points may vary within a larger range. In addition, since there is decomposition during the melting process of the substance, the melting temperature is related to the heating rate.

[0087] The term "mental illness" refers to a neurological disorder, including schizophrenia, schizoaffective psychosis, psychosis, Parkinson's disease, behavioral disorders related to dementia and psychosis, delusional disorder, acute transient psychotic disorder, depressive disorder, bipolar disorder, generalized anxiety disorder, panic disorder, obsessive-compulsive disorder, social phobia disorder, agoraphobia disorder, post-traumatic stress disorder, etc.

[0088] The term "schizophrenia" refers to a group of chronic diseases with unknown causes. Clinically, it often presents as syndromes with various symptoms, involving multiple aspects of disorders such as perception, thinking, emotion, and behavior, as well as the incoordination of mental activities. Patients are generally conscious and have basically normal intelligence, but some patients may experience impairment of cognitive function during the course of the disease.

[0089] The term "psychosis" refers to a disease manifested by disorders of mental activities such as cognition, emotion, will, and behavior to varying degrees due to the dysfunction of the brain under the influence of various biological, psychological, and social environmental factors.

[0090] The following detailed description of the invention aims to illustrate non-limiting embodiments, enabling other technicians in the field to more fully understand the technical solutions of the invention, its principles, and its practical applications, so that other technicians in the field can modify and implement the invention in many forms to best meet the requirements of specific uses.

[0091] Salts of the compound of formula I:

[0092]

[0093] In formula I, n1 and n2 are integers from 1 to 3;

[0094] R1 is selected from straight-chain or branched C1-C8 alkyl, C2-C8 alkenyl, and C2-C8 alkynyl, and the alkyl, alkenyl, and alkynyl are optionally substituted with substituents selected from halogen and C1-C8 haloalkyl;

[0095] R2 is selected from hydrogen, halogen, and C1-C8 haloalkyl;

[0096] R3, R4, R5, and R6 are each independently selected from hydrogen, halogen, and haloalkyl;

[0097] R7 is selected from straight-chain or branched C 1-8 alkyl, cycloalkyl, and R8 and R9 are each independently selected from straight-chain or branched C1-C8 alkyl, and the alkyl and cycloalkyl are optionally substituted with substituents selected from halogen and C1-C8 haloalkyl;

[0098] Z is selected from C, O, N;

[0099] Q and W are each independently selected from C and N.

[0100] In a preferred embodiment of the present invention, the salt form of the compound represented by Formula I is preferably phosphate or citrate, as shown in Formula II and Formula III:

[0101]

[0102]

[0103] y is 0.5, 1, 1.5, 2, 2.5 or 3, preferably 0.5, 1, 1.5 or 2.

[0104] In one embodiment, the phosphate or citrate of the compound represented by Formula I, wherein n1 and n2 are integers selected from 1-3. For example, n1 and n2 are each independently selected from 1, 2 and 3, such as 1, 2 or 3. In a preferred embodiment, n1 is selected from 2 and 3. In a more preferred embodiment, n1 is 1. In another preferred embodiment, n2 is 1. In a more preferred embodiment, n1 is 2. In another more preferred embodiment, n2 is 2. In a more preferred embodiment, n1 is 3. In another more preferred embodiment, n2 is 3. In a particularly preferred embodiment, n1 is 2 and n2 is 1.

[0105] In one embodiment, R1 is selected from straight-chain or branched C1-C8 alkyl, C2-C8 alkenyl and C2-C8 alkynyl, and the alkyl, alkenyl and alkynyl are each independently and optionally substituted with substituents selected from halogen and C1-C8 haloalkyl. In a preferred embodiment, R1 is straight-chain or branched C1-C8 alkyl, wherein the alkyl is optionally substituted with substituents selected from halogen and C1-C8 haloalkyl. In a more preferred embodiment, R1 is straight-chain or branched C1-C8 alkyl. In a particularly preferred embodiment, R1 is straight-chain or branched C1-C5 alkyl. In another particularly preferred embodiment, R1 is straight-chain or branched C1-C3 alkyl. In a specific embodiment, R1 is selected from methyl, ethyl, propyl, isopropyl, butyl, isobutyl, pentyl, isopentyl. In a more specific embodiment, R1 is selected from methyl, ethyl and propyl. Such as methyl, ethyl or propyl. In a particularly specific embodiment, R1 is methyl. In another particularly specific embodiment, R1 is ethyl. In yet another particularly specific embodiment, R1 is propyl.

[0106] In one embodiment, R2 is selected from hydrogen, halogen, and C1-C8 haloalkyl. In a preferred embodiment, R2 is selected from hydrogen and halogen. In a more preferred embodiment, R2 is hydrogen. In another more preferred embodiment, R2 is halogen. In a specific embodiment, R2 is selected from hydrogen, fluorine, chlorine, bromine, and iodine. In a more specific embodiment, R2 is selected from hydrogen, fluorine, chlorine, and bromine. In a more specific embodiment, R2 is fluorine. In another more specific embodiment, R2 is chlorine. In yet another more specific embodiment, R2 is bromine.

[0107] In one embodiment, R3, R4, R5, and R6 are each independently selected from hydrogen, halogen, and C1-C8 haloalkyl.

[0108] In a preferred embodiment, R3 is selected from hydrogen and halogen. In a more preferred embodiment, R3 is hydrogen. In another more preferred embodiment, R3 is halogen. In a particularly preferred embodiment, R3 is selected from fluorine, chlorine, bromine, and iodine. In a more preferred embodiment, R3 is selected from fluorine, chlorine, and bromine. In a specific embodiment, R3 is selected from hydrogen, fluorine, chlorine, and bromine. In a more specific embodiment, R3 is selected from hydrogen, fluorine, and chlorine. In a particularly specific embodiment, R3 is fluorine. In another particularly specific embodiment, R3 is chlorine.

[0109] In a preferred embodiment, R4 is selected from hydrogen and halogen. In a more preferred embodiment, R4 is hydrogen. In another more preferred embodiment, R4 is halogen. In a particularly preferred embodiment, R4 is selected from fluorine, chlorine, bromine, and iodine. In a more preferred embodiment, R4 is selected from fluorine, chlorine, and bromine. In a specific embodiment, R4 is selected from hydrogen, fluorine, chlorine, and bromine. In a more specific embodiment, R4 is selected from hydrogen, fluorine, and chlorine. In a particularly specific embodiment, R4 is fluorine. In another particularly specific embodiment, R4 is chlorine.

[0110] In a preferred embodiment, R5 is selected from hydrogen and halogen. In a more preferred embodiment, R5 is hydrogen. In another more preferred embodiment, R5 is halogen. In a particularly preferred embodiment, R5 is selected from fluorine, chlorine, bromine, and iodine. In a more preferred embodiment, R5 is selected from fluorine, chlorine, and bromine. In a specific embodiment, R5 is selected from hydrogen, fluorine, chlorine, and bromine. In a more specific embodiment, R5 is selected from hydrogen, fluorine, and chlorine. In a particularly specific embodiment, R5 is fluorine. In another particularly specific embodiment, R5 is chlorine.

[0111] In a preferred embodiment, R6 is selected from hydrogen and halogen. In a more preferred embodiment, R6 is hydrogen. In another more preferred embodiment, R6 is halogen. In a particularly preferred embodiment, R6 is selected from fluorine, chlorine, bromine and iodine. In a more preferred embodiment, R6 is selected from fluorine, chlorine and bromine. In a specific embodiment, R6 is selected from hydrogen, fluorine, chlorine and bromine. In a more specific embodiment, R6 is selected from hydrogen, fluorine and chlorine. In a particularly specific embodiment, R6 is fluorine. In another particularly specific embodiment, R6 is chlorine.

[0112] In one embodiment, R7 is selected from linear or branched C1-C8 alkyl, cycloalkyl and R8 and R9 are each independently selected from linear or branched C1-C8 alkyl, and the alkyl and cycloalkyl are optionally substituted with substituents selected from halogen and C1-C8 haloalkyl.

[0113] In a preferred embodiment, R7 is linear or branched C1-C8 alkyl. In a more preferred embodiment, R7 is linear or branched C1-C5 alkyl. In a particularly preferred embodiment, R7 is linear or branched C1-C3 alkyl. In a specific embodiment, R7 is selected from methyl, ethyl, propyl, isopropyl, butyl, isobutyl, pentyl, isopentyl. In a more specific embodiment, R7 is selected from methyl, ethyl, propyl, isopropyl, butyl, isobutyl. For example, methyl, ethyl, propyl, isopropyl, butyl or isobutyl. In a particularly specific embodiment, R7 is isopropyl.

[0114] In a preferred embodiment, R7 is cycloalkyl. In a more preferred embodiment, R7 is C3-C 10 cycloalkyl. In a particularly preferred embodiment, R7 is C3-C6 cycloalkyl. In a specific embodiment, R7 is selected from cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl. In a more specific embodiment, R7 is selected from cyclopropyl, cyclobutyl and cyclopentyl. For example, cyclopropyl, cyclobutyl or cyclopentyl.

[0115] In a preferred embodiment, R7 is wherein R8 and R9 are each independently selected from linear or branched C1-C8 alkyl. In a preferred embodiment, R7 is wherein R8 and R9 are each independently selected from linear or branched C1-C3 alkyl. In a specific embodiment, R7 is wherein R8 and R9 are each independently selected from methyl, ethyl, propyl, butyl and pentyl. In a more specific embodiment, R7 is wherein R8 and R9 are each independently selected from methyl, ethyl and propyl. For example, methyl, ethyl, n-propyl, and isopropyl. In a particularly specific embodiment, R8 and R9 are methyl.

[0116] In one embodiment, Z is selected from C, O, N. In a preferred embodiment, Z is C. In another preferred embodiment, Z is O. In yet another preferred embodiment, Z is N.

[0117] In one embodiment, Q and W are each independently selected from C, N. In a preferred embodiment, Q is N. In a preferred embodiment, Q is C. In a preferred embodiment, W is C. In a preferred embodiment, W is N.

[0118] In one embodiment, the straight-chain or branched C1-C8 alkyl group is selected from straight-chain or branched C1-C5 alkyl groups and straight-chain or branched C1-C3 alkyl groups. In a specific embodiment, the straight-chain or branched C1-C8 alkyl group, the straight-chain or branched C1-C5 alkyl group, and the straight-chain or branched C1-C3 alkyl group are each independently selected from methyl, ethyl, propyl, butyl, pentyl, and isopentyl. In a more specific embodiment, the straight-chain or branched C1-C8 alkyl group, the straight-chain or branched C1-C5 alkyl group, and the straight-chain or branched C1-C3 alkyl group are each independently selected from methyl, ethyl, propyl, and butyl.

[0119] In one embodiment, the propyl group includes but is not limited to n-propyl (n-Pr, -CH2CH2CH3) or isopropyl (i-Pr, -CH(CH3)2). The butyl group includes but is not limited to n-butyl (n-Bu, -CH2CH2CH2CH3), isobutyl (i-Bu, -CH2CH(CH3)2), sec-butyl (s-Bu, -CH(CH3)CH2CH3) or tert-butyl (t-Bu, -C(CH3)3). The pentyl group includes but is not limited to 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) or 2-methyl-1-butyl (-CH2CH(CH3)CH2CH3).

[0120] In one embodiment, the C2-C8 alkenyl is C2-C5 alkenyl. In a specific embodiment, the C2-C8 alkenyl and the C2-C5 alkenyl are each independently selected from vinyl, propenyl, butenyl, and pentenyl. In a more preferred embodiment, the C2-C8 alkenyl and the C2-C5 alkenyl are each independently selected from vinyl, propenyl, and butenyl.

[0121] In one embodiment, the propenyl includes, but is not limited to, -CH2-CH=CH2, -CH=CH-CH3. The butenyl includes, but is not limited to, -CH2-CH2-CH=CH2, -CH2-CH=CH-CH3, -CH=CH-CH2-CH3, -CH=C(CH3)2, -C(CH3)=CHCH3, -CH(CH3)CH=CH2. The pentenyl includes, but is not limited to, -CH=CHCH2CH2CH3, -CH2CH=CHCH2CH3, -CH2CH2CH=CHCH3, -CH2CH2CH2CH=CH2, -C(CH3)=CHCH2CH3, -CH(CH3)CH=CHCH3, -CH(CH3)CH2CH=CH2.

[0122] In one embodiment, the C2-C8 alkynyl is C2-C5 alkynyl. In a specific embodiment, the C2-C8 alkynyl and the C2-C5 alkynyl are each independently selected from ethynyl, propynyl, butynyl, and pentynyl. In a more specific embodiment, the C2-C8 alkynyl and the C2-C5 alkynyl are each independently selected from ethynyl, propynyl, and butynyl.

[0123] In one embodiment, the propynyl includes, but is not limited to, -H2C-C≡CH, -C≡C-CH3. The butynyl includes, but is not limited to, -H2C-CH2-C≡CH, -H2C-C≡C-CH3, H3C-CH2-C≡C-. The pentynyl includes, but is not limited to, H3C-H2C-CH2-C≡C-, -H2C-H2C-C≡C-CH3, H3C-H2C-C≡C-CH2-, (H3C)2C-C≡C-.

[0124] In one embodiment, the C1-C8 haloalkyl is C1-C5 haloalkyl. In a specific embodiment, in the C1-C8 haloalkyl and the C1-C5 haloalkyl, the C1-C8 alkyl or the C1-C5 alkyl is substituted with 1, 2, 3, or 4 halogens selected from. In a preferred embodiment, the C1-C8 alkyl or the C1-C5 alkyl is -(CH2) aCX3, where a is selected from 1, 2, 3, 4, 5, 6, and 7, and X represents a halogen. In a specific embodiment, the halogen is selected from fluorine, chlorine, bromine, and iodine. In a preferred embodiment, a is selected from 1, 2, 3, and 4. In a preferred embodiment, the halogen is fluorine.

[0125] In one embodiment, the C1-C8 haloalkyl or C1-C5 haloalkyl includes, but is not limited to, -CF3, -CCl3, -CBr3, -CI3, -CH2CF3, -CH2CCl3, -CH2CBr3, -CH2CI3, -(CH2)2CF3, -(CH2)2CCl3, -(CH2)2CBr3, -(CH2)2CI3, etc.

[0126] In one embodiment, the halogen is selected from fluorine, chlorine, bromine, and iodine. In a preferred embodiment, the halogen is selected from fluorine, chlorine, and bromine. For example, fluorine, chlorine, bromine, or iodine. In a specific embodiment, the halogen is fluorine.

[0127] In one embodiment, a phosphate or citrate of the compound represented by the general formula I, where n1 and n2 are integers from 1 to 3; R1 is methyl; R2 is selected from fluorine and hydrogen; R3, R4, R5, and R6 are each independently selected from hydrogen, fluorine, and chlorine; R7 is selected from isopropyl, cyclopropyl, isobutyl, methyl. Z is selected from C, O, and N; Q and W are each independently selected from C and N.

[0128] The present invention provides Form I of the citrate of A19006 represented by formula IV, which is characterized in that using Cu-Kα radiation, an X-ray powder diffraction pattern represented by diffraction angle 2θ ± 0.2° is obtained, which shows characteristic peaks at 10.03, 12.62, 13.69, 14.68, 16.44, 17.19, 19.10, 20.27, 21.77, wherein the error range of 2θ for each characteristic peak is ±0.2.

[0129]

[0130] In a preferred embodiment of the present invention, the X-ray powder diffraction pattern of Form I shows characteristic peaks represented by 2θ ± 0.2° at 17.67, 19.57, 21.30, 23.78, 24.03, 25.57, 26.22, wherein the error range of 2θ for each characteristic peak is ±0.2.

[0131] In a preferred embodiment of the present invention, the X-ray powder diffraction pattern of Form I shows characteristic peaks at 5.42, 10.03, 11.10, 12.62, 13.69, 14.68, 16.44, 17.19, 17.67, 18.39, 19.10, 19.57, 20.27, 21.30, 21.77, 22.50, 23.78, 24.03, 25.57, 26.22, 27.65, 28.62, 30.29, 30.56, 31.67, 32.76, 33.62, 34.04, 34.88, 35.49, 36.33, 39.02, 39.69, expressed in terms of 2θ ± 0.2°, where the error range of 2θ for each characteristic peak is ±0.2.

[0132] In a more preferred embodiment of the present invention, the melting endothermic peak of the DSC of Form I is selected from 164.7–170.0 °C, preferably 167.7 °C.

[0133] Advantageous technical effects of the present invention

[0134] Compared with the prior art, the technical solution of the present invention has the following advantages:

[0135] In vitro research results show that the compound provided by the present invention has substantially equivalent activity to pimavanserin, but the compound provided by the present invention has lower 5-HT 2A 、5-HT 2C receptor Ki values, significantly superior to pimavanserin, indicating that the compound provided by the present invention is more likely to bind to the receptor and has great clinical application value.

[0136] Experimental results show that the compound provided by the present invention acts on 5-HT 2A 、5-HT 2C receptors, has selectivity for 5-HT 2A superior to or similar to pimavanserin, has antipsychotic activity equivalent to pimavanserin, has less sedative side effects and movement deterioration side effects than pimavanserin, and has less cardiac toxicity than pimavanserin.

[0137] Compared with the compound in free base form, phosphate or citrate is beneficial to improving the solubility and hygroscopic properties of the active substance, beneficial to enhancing the bioavailability and stability of the active substance. The Form I of A19006 citrate is stable at room temperature, which is beneficial to subsequent clinical development and production development, and has good clinical application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0138] Figure 1 X-ray powder diffraction pattern of Form I of compound A19006 citrate shown in Formula IV

[0139] Figure 2 DSC spectrum of compound A19006 citrate polymorph I shown in Formula IV

[0140] Figure 3 Voltage stimulation protocol diagram for cellular hERG potassium current

[0141] Examples

[0142] The embodiments of the present invention will be 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 a limitation of the present invention. Unless otherwise specified, the ratios, percentages, etc. referred to herein are by weight.

[0143] Synthesis Examples

[0144] Example 1. Preparation of the compound shown in General Formula I

[0145] It was prepared by referring to the method described in the examples of Patent Application PCT / CN2021 / 089660.

[0146] Example 1-1. Preparation of 2-(1-methylpiperidin-4-yl)-4-{[4-(2-methylpropoxy)phenyl]methyl}-1,5-dihydro-2,4-benzodiazepin -3-one (A19020):

[0147]

[0148] Synthesis route:

[0149]

[0150] 1-1.1 Preparation of 2-[[(tert-butyldimethylsilyl)oxy]methyl]benzyl alcohol:

[0151] 1,2-Benzenedimethanol (22.0 g, 159.2 mmol), tert-butyldimethylchlorosilane (24.0 g, 159.2 mmol), triethylamine (16.1 g, 159.2 mmol), and DCM (200.00 mL) were added to a 500 mL three-necked round-bottom flask, protected by nitrogen, and stirred overnight at room temperature. Then, it was quenched with water, extracted with 3 × 100 mL of DCM, followed by washing with 200 mL of saturated brine. The DCM phase was dried over anhydrous sodium sulfate, filtered by suction, and concentrated to obtain 30 g (yield 74.64%) of a colorless oil.

[0152] 1-1.2 Preparation of 2-[[(tert-butyldimethylsilyl)oxy]methyl]benzaldehyde:

[0153] 2-[[(tert-Butyldimethylsilyl)oxy]methyl]benzyl alcohol (30.00 g, 118.8 mmol), Dess Martin (50.4 g, 118.8 mmol), and DCM (300 mL) were added to a 1 L three-necked round-bottom flask. Under nitrogen protection, the reaction was carried out at room temperature for 4 h, then quenched with water, extracted with 3 × 100 mL of DCM, washed with 200 mL of saturated brine, dried over anhydrous sodium sulfate, filtered by suction, concentrated, and purified by column chromatography with ethyl acetate / petroleum ether (1 / 35) to obtain 17 g (yield 58.62%) of a colorless oil.

[0154] Preparation of 1-1.3[(2-[[(tert-Butyldimethylsilyl)oxy]methyl]phenyl)methyl]([4-(2-methylpropoxy)phenyl]methyl])amine:

[0155] 2-[[(tert-Butyldimethylsilyl)oxy]methyl]benzaldehyde (1.00 g, 3.9 mmol), 1-(4-isobutoxyphenyl)methanamine (0.66 g, 3.9 mmol), MgSO4 (0.1 g, 0.83 mmol), and EtOH (10 mL) were added to a 50 mL three-necked round-bottom flask. After purging with nitrogen and maintaining the atmosphere, the reaction was carried out at room temperature for 3 h, then NaBH4 (0.76 g, 19.9 mmol) was added, and the mixture was stirred at room temperature for 1 h. 30 mL of water was added, and the mixture was extracted with 3 × 20 mL of ethyl acetate, washed with 30 mL of saturated brine, dried over anhydrous sodium sulfate, filtered by suction, and concentrated to obtain 1.6 g (yield 96.85%) of a colorless oil.

[0156] Preparation of 1-1.4 Benzyl-[N-2-[[(tert-Butyldimethylsilyl)oxy]methyl]benzyl]-N-[4-(2-methylpropoxy)benzyl]carbamate:

[0157] [(2-[[(tert-Butyldimethylsilyl)oxy]methyl]phenyl)methyl]([4-(2-methylpropoxy)phenyl]methyl])amine (1.6 g, 0.004 mol), tetrahydrofuran (5 mL), water (5 mL), benzyl chloroformate (0.79 g, 0.005 mol), and potassium carbonate (1.08 g, 0.008 mol) were added to a 50 mL round-bottom flask. The mixture was stirred at 50 °C for 2 h, then 30 mL of water was added, and the mixture was extracted with 3 × 20 mL of ethyl acetate, washed with 50 mL of saturated brine, dried over anhydrous sodium sulfate, filtered by suction, and concentrated to obtain 2.27 g of an oil.

[0158] Preparation of 1-1.5 Benzyl-[N-2-(hydroxymethyl)benzyl]-N-[4-(2-methylpropoxy)benzyl]carbamate:

[0159] Benzyl-[N-2-[[(tert-butyldimethylsilyl)oxy]methyl]benzyl]-N-[4-(2-methylpropoxy)benzyl]carbamate (2.27 g, 4.14 mmol), dioxane (20.00 mL), and HCl (5.2 mL) were placed in a 50 mL three-necked round-bottom flask, purged with inert nitrogen and maintained. Stir at room temperature for 1 h. Then concentrate the reaction solution and perform silica gel column chromatography with ethyl acetate / petroleum ether (1 / 5) to obtain 0.87 g (yield 48.6%) of a colorless oil.

[0160] Preparation of 1-1.6 benzyl-[N-2-(formylbenzyl)]-N-[4-(2-methylpropoxy)benzyl]carbamate:

[0161] Benzyl-[N-2-(hydroxymethyl)benzyl]-N-[4-(2-methylpropoxy)benzyl]carbamate (0.87 g, 2.0 mmol), manganese dioxide (2.62 g, 30.1 mmol), and DCM (20 mL) were placed in a 50 mL round-bottom flask, heated to 80 °C and stirred overnight, then the solid was filtered out, and the filtrate was directly concentrated to obtain 0.75 g (yield 86.61%) of an oil.

[0162] Preparation of 1-1.7 tert-butyl 4-[[2-[[[(benzyloxy)carbonyl](4-isobutoxybenzyl)amino]methyl]benzyl]amino]piperidine-1-carboxylate:

[0163] Benzyl-[N-2-(formylbenzyl)]-N-[4-(2-methylpropoxy)benzyl]carbamate (0.75 g, 1.74 mmol), 4-amino-1-tert-butoxycarbonylpiperidine (0.42 g, 2.08 mmol), magnesium sulfate (0.10 g, 0.001 mmol), and ethanol (10 mL) were added to a 50 mL three-necked round-bottom flask, purged with nitrogen and maintained, stirred at room temperature for 3 h, then NaBH4 (0.33 g, 8.69 mmol) was added, stirred at room temperature for 1 h, then 30 ml of water was added, extracted with 3 × 20 mL of ethyl acetate, washed with 50 mL of saturated brine, the ethyl acetate phase was dried over anhydrous sodium sulfate and filtered and concentrated to obtain 1.1 g of an oil.

[0164] Preparation of 1-1.8 tert-butyl 4-[[2-[[(4-isobutoxybenzyl)amino]methyl]benzyl]amino]piperidine-1-carboxylate:

[0165] tert-Butyl 4-[[2-[[[(benzyloxy)carbonyl](4-isobutoxybenzyl)amino]methyl]benzyl]amino]piperidine-1-carboxylate (1.1 g), palladium / carbon (0.1 g), methanol (5 mL), and tetrahydrofuran (5 mL) were placed in a 50 mL three-necked round-bottom flask. Hydrogen was introduced, and the mixture was stirred at room temperature for 3 h. Then, the solid was filtered off, and the filtrate was directly concentrated to obtain 0.65 g of an oily substance.

[0166] 1 - 1.9 Preparation of 2-(tert-butoxycarbonylpiperidin)-4-[[4-(2-methylpropoxy)phenyl]methyl]-1,5-dihydro-2,4-benzodiazepin -3-one:

[0167] tert-Butyl 4-[[2-[[(4-isobutoxybenzyl)amino]methyl]benzyl]amino]piperidine-1-carboxylate (0.65 g, 1.34 mmol) and tetrahydrofuran (10 mL) were added to a 50 mL round-bottom flask. The flask was purged and maintained with inert nitrogen. BTC (0.16 g, 0.54 mmol) was added at -40 °C, and the reaction was kept at this temperature for 1 h. Then, 30 mL of water was added, and the mixture was extracted with 3 × 20 mL of ethyl acetate. The extract was washed with 50 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain 0.78 g of an oily substance.

[0168] 1 - 1.10 Preparation of 2-(piperidin-4-yl)-4-[[4-(2-methylpropoxy)phenyl]methyl]-1,5-dihydro-2,4-benzodiazepin -3-one:

[0169] 2-(tert-Butoxycarbonylpiperidin)-4-[[4-(2-methylpropoxy)phenyl]methyl]-1,5-dihydro-2,4-benzodiazepin -3-one (0.78 g, 1.53 mmol), DCM (10 mL), and HCl / dioxane 4 M (0.22 g, 1.6 mmol) were added to a 50 mL round-bottom flask. The mixture was stirred at room temperature for 1 h, and then the reaction solution was directly concentrated to obtain 0.66 g of an oily substance.

[0170] 1 - 1.11 Preparation of 2-(1-methylpiperidin-4-yl)-4-[[4-(2-methylpropoxy)phenyl]methyl]-1,5-dihydro-2,4-benzodiazepin -3-one:

[0171] 2-(Piperidin-4-yl)-4-[[4-(2-methylpropoxy)phenyl]methyl]-1,5-dihydro-2,4-benzodiazepin -3-one (0.66 g, 1.6 mmol), formaldehyde (0.1 g, 3.2 mmol), triethylamine (0.66 g, 6.4 mmol), magnesium sulfate (0.02 g, 0.16 mmol), methanol (10 mL) were added to a 50 mL round-bottom flask, stirred at room temperature for 3 h, then NaBH4 (0.31 g, 8.1 mmol) was added, stirred at room temperature for 1 h, then 30 mL of water was added, extracted with 3×20 mL of ethyl acetate, washed with 30 mL of saturated brine, the ethyl acetate phase was dried over anhydrous sodium sulfate and filtered and concentrated, and finally purified by an Intel Flash-1 chromatographic column to obtain 11.6 mg (yield 1.7%) of a pale yellow oil. 1 H NMR (400 MHz, Methanol-d4) δ 7.40 - 7.19 (m, 5H), 7.03 (d, J = 7.3 Hz, 1H), 6.91–6.82 (m, 2H), 4.51 (s, 2H), 4.46 (s, 2H), 4.41 (s, 2H), 4.34 (tt, J = 12.2, 3.9 Hz, 1H), 3.74 (d, J = 6.5 Hz, 2H), 3.64 - 3.56 (m, 2H), 3.21 - 3.04 (m, 2H), 2.91 (s, 3H), 2.32 - 2.17 (m, 2H), 2.07 (dq, J = 13.3, 6.6 Hz, 1H), 1.97 (t, J = 15.8 Hz, 2H), 1.04 (d, J = 6.7 Hz, 6H), LCMS (ES, m / z): 422 [M+H] + .

[0172] Example 1-2: Preparation of 7-fluoro-2-(1-methylpiperidin-4-yl)-4-[[4-(2-methylpropoxy)phenyl]methyl]-1,5-dihydro-2,4-benzodiazepin -3-one (A19001):

[0173]

[0174] Synthetic route:

[0175]

[0176] 1-2.1 Preparation of methyl 2-[(tert-butoxycarbonyl)aminomethyl]-5-fluorobenzoate:

[0177] Methyl 2-cyano-5-fluorobenzoate (1.8 g, 10.05 mmol), Raney-Ni (0.5 g), Boc2O (2.63 g, 12.06 mmol), NaHCO3 (1.69 g, 20.1 mmol), and THF (20 mL) were added to a 50 mL three-necked round-bottom flask. Hydrogen was introduced, and the mixture was stirred at 50 °C for 48 h. Then the reaction mixture was filtered, and the filtrate was directly concentrated to obtain 2 g (70.26%) of a solid.

[0178] Preparation of 2-[(tert-butoxycarbonyl)aminomethyl]-5-fluorobenzoic acid:

[0179] Methyl 2-[(tert-butoxycarbonyl)aminomethyl]-5-fluorobenzoate (2.0 g, 7.06 mmol), sodium hydroxide (1.41 g, 35.3 mmol), and THF / H2O (10 / 10 mL) were placed in a 50 mL three-necked round-bottom flask and stirred at room temperature overnight. Then the pH was adjusted to 5 with 3N HCl, and the mixture was extracted with 3 × 20 mL of EA, washed with 30 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain 1.2 g (yield 63.13%) of a solid.

[0180] Preparation of tert-butyl [4-fluoro-2-[(4-isobutoxybenzyl)carbamoyl]benzyl]carbamate:

[0181] 2-[(tert-butoxycarbonyl)aminomethyl]-5-fluorobenzoic acid (1.2 g, 4.4 mmol), 4-isobutoxybenzylamine (0.88 g, 4.9 mmol), HATU (2.2 g, 5.8 mmol), DIEA (1.15 g, 8.9 mmol), and DMF (20 mL) were placed in a 50 mL three-necked round-bottom flask and stirred at room temperature overnight. Then 30 mL of water was added, and the mixture was extracted with 3 × 20 mL of EA, washed with 30 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated. Silica gel column chromatography with ethyl acetate / petroleum ether (1 / 3) was used to obtain 1 g (yield 52.12%) of a solid.

[0182] Preparation of 2-(aminomethyl)-5-fluoro-N-(4-isobutoxybenzyl)benzamide:

[0183] Prepared according to the method of Example 1-1.10 to obtain 1 g of an oily substance.

[0184] Preparation of 5-fluoro-N-(4-isobutoxybenzyl)-2-[[(1-methylpiperidin-4-yl)amino]methyl]benzamide:

[0185] 2-(Aminomethyl)-5-fluoro-N-(4-isobutoxybenzyl)benzamide (1 g, 3.02 mmol), 1-methylpiperidin-4-one (0.41 g, 3.6 mmol), NaBH3CN (0.38 g, 6.05 mmol), EtOH (10 mL) and HOAc (1 mL) were placed in a 50 mL round-bottom flask and stirred overnight at room temperature. Then 30 mL of saturated NaHCO3 solution was added, and the mixture was extracted with 3×20 mL of EA, washed with 30 mL of saturated brine, dried over anhydrous sodium sulfate and filtered and concentrated under reduced pressure. Silica gel column chromatography with dichloromethane / methanol (10 / 1) was used to obtain 0.75 g (yield 57.96%) of a solid.

[0186] Preparation of 1-2.6 N-[4-fluoro-2-[(4-isobutoxybenzyl)amino]methyl]benzyl}-1-methylpiperidin-4-amine:

[0187] 5-Fluoro-N-(4-isobutoxybenzyl)-2-[[(1-methylpiperidin-4-yl)amino]methyl]benzamide (0.3 g, 0.702 mmol) and BH3-THF (10.00 mL) were placed in a 50 mL round-bottom flask and stirred overnight under reflux. Then it was quenched with 2N HCl, EA (30 mL) was added, the aqueous phase was extracted with EA (2×20 mL), then the pH of the aqueous phase was adjusted to 10 with 15% NaOH solution, and extracted with DCM. The solvent was removed, and then purified by reverse chromatography to obtain 0.1 g (yield 34.46%) of a white solid.

[0188] 1-2.7 7-Fluoro-2-(1-methylpiperidin-4-yl)-4-[[4-(2-methylpropoxy)phenyl]methyl]-1,5-dihydro-2,4-benzodiaz -3-one Preparation:

[0189] Prepared according to the method of Example 1-1.9 to obtain 2.8 mg (yield 2.63%) of a white solid. H-NMR (400 MHz, Methanol-d4): δ 7.34 (dd, J = 8.4, 5.4 Hz, 1H), 7.25 - 7.17 (m, 2H), 7.00 (td, J = 8.6, 2.7 Hz, 1H), 6.91 - 6.82 (m, 2H), 6.77 (dd, J = 9.1, 2.7 Hz, 1H), 4.47 (d, J = 15.2 Hz, 4H), 4.39 (s, 2H), 4.43–4.27 (m, 1H), 3.74 (d, J = 6.4 Hz, 2H), 3.64 - 3.56 (m, 2H), 3.21–3.10 (m, 2H), 2.90 (s, 3H), 2.32 (dd, J = 13.4, 4.0 Hz, 1H), 2.25 (dd, J = 13.1, 4.1 Hz, 1H), 2.06 (dp, J = 13.3, 6.7 Hz, 1H), 1.95 (d, J = 13.9 Hz, 2H), 1.04 (d, J = 6.7 Hz, 6H); LCMS (ES, m / z): 440 [M+H] + .

[0190] Example 1-3: Preparation of 7-chloro-2-(1-methylpiperidin-4-yl)-4-[[4-(2-methylpropoxy)phenyl]methyl]-1,5-dihydro-2,4-benzodiazepin -3-one (A190017):

[0191]

[0192] Synthetic route:

[0193]

[0194] 1-3.1 Preparation of methyl 2-[(tert-butoxycarbonyl)aminomethyl]-4-chlorobenzoate:

[0195] Prepared according to the method of Example 1-2.1 to obtain 1.58 g (103.10%) of a white solid.

[0196] 1-3.2 Preparation of 2-[(tert-butoxycarbonyl)aminomethyl]-4-chlorobenzoic acid:

[0197] Prepared according to the method of Example 1-2.2 to obtain 1.12 g (74.37%) of a white solid.

[0198] 1-3.3 Preparation of 2-[(tert-butoxycarbonyl)aminomethyl]-4-chloro-4-(piperidine-1-carboxylic acid benzyl ester)benzamide:

[0199] Prepared according to the method of Example 1-2.3 to obtain 2 g (101.63%) of a yellow oil.

[0200] 1-3.4 Preparation of 2-(aminomethyl)-4-chloro-4-(benzyl piperidine-1-carboxylate) benzamide:

[0201] Prepared according to the method of Example 1-1.10 to obtain 2 g (124.91%) of a white solid.

[0202] 1-3.5 Preparation of 2-(2-methylpropoxy)-benzylamine-4-chloro-4-(benzyl piperidine-1-carboxylate) benzamide:

[0203] Prepared according to the method of Example 1-2.5 to obtain 1 g (35.62%) of a yellow oil.

[0204] 1-3.6 Preparation of 2-(2-methylpropoxy)-benzylamine-4-chloro-4-(benzyl piperidine-1-carboxylate) benzylamine:

[0205] Prepared according to the method of Example 1-2.6 to obtain 0.22 g (22.56%) of a yellow oil.

[0206] 1-3.7 Preparation of 7-chloro-2-(benzyl piperidine-1-carboxylate)-4-[[4-(2-methylpropoxy)phenyl]methyl]-1,5-dihydro-2,4-benzodiazepin -3-one:

[0207] Prepared according to the method of Example 1-1.9 to obtain 0.21 g (yield 91.15%) of a yellow oil.

[0208] 1-3.8 Preparation of 7-chloro-2-(1-methylpiperidin-4-yl)-4-[[4-(2-methylpropoxy)phenyl]methyl]-1,5-dihydro-2,4-benzodiazepin -3-one:

[0209] 7-chloro-2-(benzyl piperidine-1-carboxylate)-4-[[4-(2-methylpropoxy)phenyl]methyl]-1,5-dihydro-2,4-benzodiazepin -3-one (0.21 g, 0.365 mmol), Pd / C (77.58 mg, 0.729 mmol), HCHO (43.78 mg, 1.46 mmol), MeOH (3 mL) were placed in a 50 mL round-bottom flask, stirred at room temperature for 10 h, filtered by suction, the filtrate was concentrated, and purified by preparative liquid chromatography to obtain 2.4 mg (yield 1.44%) of a white solid. 11H NMR (400 MHz, DMSO-d6) δ 9.62 (s, 1H), 7.33 (dd, J = 8.0, 2.2 Hz, 1H), 7.28 - 7.18 (m, 6H), 6.90 - 6.83 (m, 3H), 4.37 - 4.30 (m, 8H), 4.17 (d, J = 12.6 Hz, 2H), 3.72 (d, J = 6.5 Hz, 3H), 3.05 (d, J = 10.8 Hz, 2H), 2.76 (d, J = 4.7 Hz, 4H), 2.38 (s, 4H), 2.18 - 2.08 (m, 3H), 2.00 (dt, J = 13.4, 6.7 Hz, 1H), 1.75 (d, J = 13.3 Hz, 3H), 1.24 (s, 1H), 0.98 (d, J = 6.7 Hz, 9H); LCMS (ES, m / z): 456 [M+H] + .

[0210] Examples 1 - 4: Preparation of 8 - fluoro - 2 - (1 - methylpiperidin - 4 - yl) - 4 - [[4 - (2 - methylpropoxy)phenyl]methyl] - 1,5 - dihydro - 2,4 - benzodiazepin - 3 - one (A19005): - 3 - one preparation (A19005):

[0211]

[0212] Synthesis route:

[0213]

[0214] 1 - 4.1 Preparation of tert - butyl 4 - fluoro - 2 - [[(1 - methylpiperidin - 4 - yl)carbamoyl]benzyl]carbamate:

[0215] Prepared according to the method of Example 1 - 2.3, 8 g (84.21%) of an oil was obtained.

[0216] 1 - 4.2 Preparation of 2 - (aminomethyl) - 5 - fluoro - N - (1 - methylpiperidin - 4 - yl)benzamide:

[0217] Prepared according to the method of Example 1 - 1.10, 2 g of an oil was obtained.

[0218] 1 - 4.3 Preparation of 5 - fluoro - 2 - [[(4 - isobutoxybenzyl)amino]methyl] - N - (1 - methylpiperidin - 4 - yl)benzamide:

[0219] Prepared according to the method of Example 1 - 2.5, 0.65 g (20.17%) of a yellow oil was obtained.

[0220] Preparation of 1-4.4N-[5-fluoro-2-[[(4-isobutoxybenzyl)amino]methyl]benzyl]-1-methylpiperidin-4-amine:

[0221] Prepared according to the method of Example 1-2.6, obtaining 0.25 g (12.31%) of a yellow oil.

[0222] 1-4.5 Preparation of 8-fluoro-2-(1-methylpiperidin-4-yl)-4-[[4-(2-methylpropoxy)phenyl]methyl]-1,5-dihydro-2,4-benzodiazepin -3-one:

[0223] Prepared according to the method of Example 1-1.9, obtaining 35 mg (yield 13.17%) of a white solid. H-NMR (400 MHz, Methanol-d4) δ 7.24 - 7.17 (m, 2H), 7.11 (dd, J = 9.0, 2.6 Hz, 1H), 7.03 (dd, J = 8.4, 5.5 Hz, 1H), 6.95 (td, J = 8.6, 2.6 Hz, 1H), 6.90 - 6.83 (m, 2H), 4.52 - 4.36 (m, 7H), 4.31 (tt, J = 12.3, 3.9 Hz, 1H), 3.73 (d, J = 6.5 Hz, 2H), 3.60 (d, J = 12.4 Hz, 2H), 3.40 (s, 0H), 3.15 (t, J = 12.7 Hz, 2H), 2.91 (s, 3H), 2.25 (qd, J = 13.3, 4.0 Hz, 2H), 2.06 (dt, J = 13.3, 6.6 Hz, 1H), 1.96 (d, J = 13.8 Hz, 2H), 1.04 (d, J = 6.7 Hz, 6H); LCMS (ES, m / z): 440 [M+H] + .

[0224] Example 1-5: Preparation of 8-fluoro-2-(1-methylpiperidin-4-yl)-4-[[4-(2-methylpropylamino)phenyl]methyl]-1,5-dihydro-2,4-benzodiazepin -3-one (A19007):

[0225]

[0226] Synthetic route:

[0227]

[0228] 1-5.1 Preparation of methyl 2-[(tert-butoxycarbonyl)aminomethyl]-5-fluorobenzoate:

[0229] Prepared according to the method of Example 1-2.1, obtaining 4.7 g of an oil.

[0230] 1-5.2 Preparation of 2-[(tert-butoxycarbonyl)aminomethyl]-5-fluorobenzoic acid:

[0231] Prepared according to the method of Example 1-2.2, and 4.0 g (89.54%) of an oily substance was obtained.

[0232] 1-5.3 Preparation of 2-[(tert-butoxycarbonyl)aminomethyl]-5-fluoro-N-(4-(benzyl piperidine-1-carboxylate))benzamide was prepared according to the method of Example 1-2.3, and 4.6 g of an oily substance was obtained.

[0233] 1-5.4 Preparation of 2-(aminomethyl)-5-fluoro-N-(4-(benzyl piperidine-1-carboxylate))benzamide:

[0234] Prepared according to the method of Example 1-1.10, and 5.6 g of an oily substance was obtained.

[0235] 1-5.5 Preparation of benzyl 4-[2-[(4-bromobenzamido)methyl]-5-fluorobenzamido]piperidine-1-carboxylate:

[0236] Prepared according to the method of Example 1-2.3, and 3.2 g (38.75%) of a white solid was obtained.

[0237] 1-5.6 Preparation of benzyl 4-[5-fluoro-2-[[4-(isobutylamino)benzamido]methyl]benzamido]piperidine-1-carboxylate:

[0238] Benzyl 4-[2-[(4-bromobenzamido)methyl]-5-fluorobenzamido]piperidine-1-carboxylate (1 g, 1.76 mmol), isobutylamine (0.193 g, 2.64 mmol), sodium tert-butoxide (0.34 g, 3.52 mmol), Ruphos (82.1 mg, 0.176 mmol), Pd2(dba)3 (0.161 mg, 0.176 mmol), and toluene (13 mL) were placed in a 50 mL round-bottom flask, heated to 80 °C and stirred for 1 h, filtered by suction, the filtrate was concentrated, and silica gel column chromatography (PE / EA = 3:1) was performed to obtain 0.797 g (yield 80.8%) of a yellow solid.

[0239] 1-5.7 Preparation of benzyl 4-[[5-fluoro-2-[[(4-(isobutylamino)benzyl]amino]methyl]benzyl(amino)piperidine-1-carboxylate:

[0240] Prepared according to the method of 2.6 in Example 2, and 260 mg of an off-white solid (34.33%) was obtained.

[0241] 1-5.8 Preparation of 8-fluoro-2-(benzyl piperidine-1-carboxylate)-4-[[4-(2-methylpropylamino)phenyl]methyl]-1,5-dihydro-2,4-benzodiazepin -3-one:

[0242] Prepared according to the method of Example 1-1.9 to obtain 0.26 g of an oily substance.

[0243] 1-5.9 Preparation of 8-fluoro-2-(1-methylpiperidin-4-yl)-4-[[4-(2-methylpropylamino)phenyl]methyl]-1,5-dihydro-2,4-benzodiazepin -3-one:

[0244] Prepared according to the method of Example 1-3.8 to obtain 4.1 mg (yield 2.61%) of a white solid. 1 H NMR (400 MHz, Methanol-d4) δ 8.48 (s, 1H, FA), 7.10 - 6.98 (m, 4H), 6.93 (td, J = 8.6, 2.7 Hz, 1H), 6.58 (d, 2H), 4.42 (s, 4H), 4.36 (s, 2H), 4.30 - 4.18 (m, 1H), 3.46 - 3.39 (m, 2H), 2.96 - 2.83 (m, 4H), 2.77 (s, 3H), 2.25 - 2.05 (m, 2H), 1.96 - 1.84 (m, 3H), 1.36 - 1.30 (m, 1H), 0.99 (d, J = 6.6 Hz, 6H); LCMS (ES, m / z): 439 [M+H] + .

[0245] The general synthesis formula of compounds A19006, A19008, A19009, A19011, A19012, A19015 is as follows:

[0246]

[0247] Example 1-6: Preparation of 7-fluoro-2-[(4-cyclopropoxyphenyl)methyl]-4-(1-methylpiperidin-4-yl)-1,5-dihydro-2,4-benzodiazepin -3-one (A19006):

[0248]

[0249] 1-6.1 Preparation of 2-[(tert-butoxycarbonyl)aminomethyl]-5-fluoro-N-(4-(benzyl piperidine-1-carboxylate))benzamide:

[0250] Prepared according to the method of Example 1-2.3 to obtain 2.3 g (63.77%) of a yellow solid.

[0251] Preparation of Benzyl 4-[2-(aminomethyl)-5-fluorobenzamido]piperidine-1-carboxylate Hydrochloride:

[0252] Prepared according to the method of Example 1-1.10 to obtain 1.9 g of an oily substance.

[0253] Preparation of Benzyl 4-[2-[[(4-Cyclopropoxybenzyl)amino]methyl]-5-fluorobenzamido]piperidine-1-carboxylate:

[0254] Prepared according to the method of Example 1-1.7 to obtain 1.3 g of an oily substance.

[0255] Preparation of Benzyl 4-[2-[[(4-Cyclopropoxybenzyl)amino]methyl]-5-fluorobenzylamino]piperidine-1-carboxylate:

[0256] Prepared according to the method of Example 1-2.6 to obtain 0.23 g (18.17%) of a yellow oily substance.

[0257] 1-6.5 Preparation of Benzyl 4-[4-(4-Cyclopropoxybenzyl)-8-fluoro-3-oxo-1,3,4,5-tetrahydro-2H-benzo[e][1,3]diazin -2-yl]piperidine-1-carboxylate:

[0258] Prepared according to the method of Example 1-1.9 to obtain 0.2 g of an oily substance.

[0259] 1-6.6 Preparation of 7-Fluoro-2-[(4-cyclopropoxyphenyl)methyl]-4-(1-methylpiperidin-4-yl)-1,5-dihydro-2,4-benzodiazepin -3-one:

[0260] Prepared according to the method of Example 1-3.8 to obtain 37.9 mg (yield 24.32%) of a brown solid. 11H NMR (400 MHz, DMSO-d6) δ 7.30 - 7.19 (m, 3H), 7.11 (dd, J = 8.4, 5.7 Hz, 1H), 7.05 - 6.94 (m, 3H), 4.36 (d, J = 7.1 Hz, 4H), 4.29 (s, 2H), 3.96 (tt, J = 11.4, 4.0 Hz, 1H), 3.80 (tt, J = 6.0, 3.0 Hz, 1H), 2.86 - 2.78 (m, 2H), 2.16 (s, 3H), 1.95 - 1.85 (m, 2H), 1.89 - 1.75 (m, 2H), 1.45 (dd, J = 11.0, 3.9 Hz, 2H), 0.81 - 0.67 (m, 2H), 0.67 - 0.59 (m, 2H); LCMS (ES, m / z): 424 [M+H] + .

[0261] Examples 1 - 7: Preparation of 8 - fluoro - 2 - (1 - methylpiperidin - 4 - yl) - 4 - [[4 - (2 - methylpropyl)phenyl]methyl] - 1,5 - dihydro - 2,4 - benzodiazepin -3 - one (A19008):

[0262]

[0263] 1 - 7.1 Preparation of benzyl 4 - [5 - fluoro - 2 - [(4 - isobutylbenzyl)amino]methyl]benzamido]piperidine - 1 - carboxylate:

[0264] Prepared according to the method of Example 1 - 1.7, obtaining 2.3 g (yield 61.76%) of a yellow oil.

[0265] 1 - 7.2 Preparation of benzyl 4 - [[5 - fluoro - 2 - [(4 - isobutylbenzyl)amino]methyl]benzyl]amino)piperidine - 1 - carboxylate:

[0266] Prepared according to the method of Example 1 - 2.6, obtaining 0.69 g of a yellow oil.

[0267] 1 - 7.3 Preparation of 8 - fluoro - 2 - (4 - (piperidine - 1 - carboxylate benzyl)) - 4 - [[4 - (2 - methylpropyl)phenyl]methyl] - 1,5 - dihydro - 2,4 - benzodiazepin -3 - one:

[0268] Prepared according to the method of Example 1 - 1.9, obtaining 0.7 g of a yellow oil.

[0269] 1 - 7.4 8 - fluoro - 2 - (1 - methylpiperidin - 4 - yl) - 4 - [[4 - (2 - methylpropyl)phenyl]methyl] - 1,5 - dihydro - 2,4 - benzodiazepin Preparation of -3-one:

[0270] Prepared according to the method of Example 1-3.8 to obtain 21 mg (yield 3.85%) of a yellow solid. 1 H NMR (400 MHz, DMSO-d6) δ 10.87 (s, 1H), 7.23 - 6.96 (m, 7H), 4.43 (d, J = 7.8 Hz, 4H), 4.34 (s, 3H), 3.86 (s, 1H), 3.45 - 3.37 (m, 2H), 3.17 (s, 1H), 3.11 - 2.95 (m, 2H), 2.71 (d, J = 4.8 Hz, 3H), 2.44 - 2.27 (m, 4H), 1.80 (dp, J = 13.5, 6.8 Hz, 1H), 1.72 - 1.59 (m, 2H), 0.85 (d, J = 6.6 Hz, 6H); LCMS (ES, m / z): 424 [M + H] + .

[0271] Example 1-8: Preparation of 8-fluoro-2-(1-methylpiperidin-4-yl)-4-[(4-methoxyphenyl)methyl]-1,5-dihydro-2,4-benzodiazepin -3-one (A19009):

[0272]

[0273] 1-8.1 Preparation of benzyl 4-[5-fluoro-2-[[(4-methoxybenzyl)amino]methyl]benzamide]piperidine-1-carboxylate:

[0274] Prepared according to the method of Example 1-1.7 to obtain 0.48 g of a yellow oil.

[0275] 1-8.2 Preparation of benzyl 4-[5-fluoro-2-[[(4-methoxybenzyl)amino]methyl]benzyl]aminopiperidine-1-carboxylate:

[0276] Prepared according to the method of Example 1-2.6 to obtain 0.13 g of a yellow oil.

[0277] 1-8.3 Preparation of 8-fluoro-2-(4-(piperidine-1-carboxylic acid benzyl ester))-4-[(4-methoxyphenyl)methyl]-1,5-dihydro-2,4-benzodiazepin -3-one:

[0278] Prepared according to the method of Example 1-1.9 to obtain 0.09 g of a yellow oil.

[0279] 1-8.4 8-fluoro-2-(1-methylpiperidin-4-yl)-4-[(4-methoxyphenyl)methyl]-1,5-dihydro-2,4-benzodiazepin Preparation of 1,5-dihydro-2,4-benzodiazepin-3-one:

[0280] Prepared according to the method of Example 1-3.8 to obtain 9 mg (yield 13.2%) of a white solid. 1 H NMR (400 MHz, DMSO-d6) δ 7.25 (dd, J = 11.0, 7.5 Hz, 3H), 7.10 (dd, J = 8.5, 5.7 Hz, 1H), 7.01 (td, J = 8.7, 2.7 Hz, 1H), 6.87 (d, J = 8.4 Hz, 2H), 4.36 (d, J = 7.2 Hz, 4H), 4.28 (s, 2H), 4.03 - 3.92 (m, 1H), 3.73 (s, 3H), 2.85 (d, J = 9.8 Hz, 2H), 2.20 (s, 3H), 1.96 (s, 2H), 1.91 - 1.77 (m, 2H), 1.46 (d, J = 11.2 Hz, 2H); LCMS (ES, m / z): 398 [M+H] + .

[0281] Example 1-9: Preparation of 8-fluoro-2-(1-methylpiperidin-4-yl)-4-[[(2-fluoro-4-isopropoxyphenyl)methyl]-1,5-dihydro-2,4-benzodiazepin-3-one (A19011): 1-9.1 Preparation of benzyl 4-[5-fluoro-2-[(2-fluoro-4-isopropoxybenzyl)amino]methyl]benzamide piperidine-1-carboxylate:

[0282]

[0283] Prepared according to the method of Example 1-1.7 to obtain 0.88 g of a yellow oil.

[0284] Prepared according to the method of Example 1-1.7 to obtain 0.88 g of a yellow oil.

[0285] 1-9.2 Preparation of benzyl 4-[5-fluoro-2-[(2-fluoro-4-isopropoxybenzyl)amino]methyl]benzyl]amino]piperidine-1-carboxylate:

[0286] Prepared according to the method of Example 1-2.6 to obtain 0.117 g of a yellow oil.

[0287] 1-9.3 Preparation of 8-fluoro-2-(4-(piperidine-1-carboxylic acid benzyl ester))-4-[[(2-fluoro-4-isopropoxyphenyl)methyl]-1,5-dihydro-2,4-benzodiazepin-3-one: Prepared according to the method of Example 1-1.9 to obtain 0.09 g of a yellow oil.

[0288] Prepared according to the method of Example 1-1.9 to obtain 0.09 g of a yellow oil.

[0289] 1-9.4 Preparation of 8-fluoro-2-(1-methylpiperidin-4-yl)-4-[[(2-fluoro-4-isopropoxyphenyl)methyl]-1,5-dihydro-2,4-benzodiazepin -3-one:

[0290] Prepared according to the method of Example 1-3.8 to obtain 5.3 mg (yield 7.5%) of an orange solid. 1 H NMR (400 MHz, DMSO-d6) δ 9.54 (s, 1H), 7.22 - 7.00 (m, 4H), 6.76 (dd, J = 12.4, 2.5 Hz, 1H), 6.69 (dd, J = 8.5, 2.5 Hz, 1H), 4.60 (p, J = 6.0 Hz, 1H), 4.42 (s, 2H), 4.36 (d, J = 16.6 Hz, 4H), 2.76 (d, J = 4.7 Hz, 3H), 2.14 - 2.04 (m, 1H), 1.74 (d, J = 14.9 Hz, 2H), 1.25 (d, J = 5.9 Hz, 7H); LCMS (ES, m / z): 444 [M + H] + .

[0291] Example 1-10: Preparation of 8-fluoro-2-(1-methylpiperidin-4-yl)-4-{[(3-fluoro-4-isopropoxyphenyl)methyl]-1,5-dihydro-2,4-benzodiazepin -3-one (A19012):

[0292]

[0293] 1-10.1 Preparation of benzyl 4-[5-fluoro-2-[(3-fluoro-4-isopropoxybenzyl)amino]methyl]benzamide]piperidine-1-carboxylate:

[0294] Prepared according to the method of Example 1-1.7 to obtain 1.0 g of a yellow oil.

[0295] 1-10.2 Preparation of benzyl 4-[5-fluoro-2-[(3-fluoro-4-isopropoxybenzyl)amino]methyl]benzyl]amino]piperidine-1-carboxylate:

[0296] Prepared according to the method of Example 1-2.6 to obtain 0.118 g of a yellow oil.

[0297] 1-10.3 Preparation of 8-fluoro-2-(4-(piperidine-1-carboxylic acid benzyl ester))-4-[[(3-fluoro-4-isopropoxyphenyl)methyl]-1,5-dihydro-2,4-benzodiazepin -3-one:

[0298] Prepared according to the method of Example 1-1.9 to obtain 0.11 g of a yellow oil.

[0299] 1-10.4 8-Fluoro-2-(1-methylpiperidin-4-yl)-4-[[(3-fluoro-4-isopropoxyphenyl)methyl]-1,5-dihydro-2,4-benzodiazepin -3-one Preparation:

[0300] Prepared according to the method of Example 1-3.8 to obtain 4.3 mg (yield 4.9%) of a yellow oil. 1 H NMR (400 MHz, DMSO-d6) δ 9.16 (s, 1H), 7.18 (dd, J = 8.3, 5.6 Hz, 1H), 7.15 - 7.01 (m, 5H), 4.58 (p, J = 6.0 Hz, 1H), 4.40 - 4.32 (m, 6H), 4.16 (s, 1H), 2.78 (d, J = 4.4 Hz, 3H), 2.44 (s, 12H), 2.05 (q, J = 12.9, 12.3 Hz, 2H), 1.78 (d, J = 13.5 Hz, 2H), 1.27 (d, J = 6.0 Hz, 6H), 1.24 (s, 1H); LCMS (ES, m / z): 444 [M+H] + .

[0301] Example 1-11: Preparation of 7-Fluoro-2-[(6-isopropoxypyridin-3-yl)methyl]-4-(1-methylpiperidin-4-yl)-1,5-dihydro-2,4-benzodiazepin -3-one Preparation (A19013):

[0302]

[0303] Synthesis Route:

[0304]

[0305] 1-11.1 Preparation of 2-[(6-isopropoxypyridin-3-yl)]-methyl-amino-methyl-5-fluoro-4-(piperidine-1-carboxylic acid benzyl ester)benzamide:

[0306] Prepared according to the method of Example 1-1.7 to obtain 2.4 g of a yellow oil.

[0307] 1-11.2 Preparation of 2-[4-(4-isopropoxypyridin-3-yl)]-methyl-amino-methyl-5-fluoro-4-(piperidine-1-carboxylic acid benzyl ester)benzylamine:

[0308] Prepared according to the method of Example 1-2.6 to obtain 0.38 g of a yellow oil.

[0309] 1-11.3 Preparation of 8-fluoro-2-(4-(benzyl piperidine-1-carboxylate))-4-[[(6-isopropoxypyridin-3-yl)methyl]]-1,5-dihydro-2,4-benzodiazepin-3-one: Prepared according to the method of Example 1-1.9 to obtain 0.37 g of a yellow oil.

[0310] Prepared according to the method of Example 1-1.9 to obtain 0.37 g of a yellow oil.

[0311] 1-11.4 Preparation of 8-fluoro-2-(1-methylpiperidin-4-yl)-4-[[(6-isopropoxypyridin-3-yl)methyl]]-1,5-dihydro-2,4-benzodiazepin-3-one: Prepared according to the method of Example 1-3.8 to obtain 50.6 mg (yield 17.3%) of a white solid.

[0312] Prepared according to the method of Example 1-3.8 to obtain 50.6 mg (yield 17.3%) of a white solid. 1 H NMR (400 MHz, DMSO-d6) δ 1 H NMR (400 MHz, DMSO-d6) δ 8.09 (s, 1H), 7.62 (d, J = 8.6 Hz, 1H), 7.27 (d, J = 9.3 Hz, 1H), 7.15 (t, J = 7.1 Hz, 1H), 7.02 (t, J = 8.3 Hz, 1H), 6.67 (d, J = 8.6 Hz, 1H), 5.21 (dt, J = 13.0, 6.6 Hz, 1H), 4.39 - 4.31 (m, 6H), 3.98 (d, J = 12.3 Hz, 1H), 2.82 (d, J = 10.3 Hz, 2H), 2.17 (s, 3H), 1.88 (dt, J = 28.7, 12.2 Hz, 4H), 1.48 - 1.40 (m, 2H), 1.27 (d, J = 6.1 Hz, 6H); LCMS (ES, m / z): 427 [M + H] + .

[0313] Example 1-12: Preparation of 8-fluoro-2-(1-methylpyrrol-3-yl)-4-[[(4-isobutoxyphenyl)methyl]]-1,5-dihydro-2,4-benzodiazepin-3-one (A19015): Preparation of 2-[(4-isobutoxyphenyl-3-yl)]-methyl-amino-methyl-5-fluoro-4-(pyrrole-1-carboxylic acid benzyl ester) benzamide:

[0314]

[0315] 1-12.1 Preparation of 2-[(4-isobutoxyphenyl-3-yl)]-methyl-amino-methyl-5-fluoro-4-(pyrrole-1-carboxylic acid benzyl ester) benzamide:

[0316] Prepared according to the method of Example 1-1.7 to obtain 3.3 g of a yellow oil.

[0317] Preparation of 1-12.2 2-[4-(4-isobutoxyphenyl-3-yl)]-methyl-amino-methyl-5-fluoro-4-(benzyl piperidine-1-carboxylate) benzylamine:

[0318] Prepared according to the method of Example 1-2.6 to obtain 0.32 g of a colorless oil.

[0319] Preparation of 1-12.3 8-fluoro-2-(3-(benzyl pyrrole-1-carboxylate))-4-[[(4-isobutoxyphenyl)methyl]]-1,5-dihydro-2,4-benzodiazepin -3-one:

[0320] Prepared according to the method of Example 1-1.9 to obtain 0.31 g of a yellow oil.

[0321] Preparation of 1-12.4 8-fluoro-2-(1-methylpyrrol-3-yl)-4-[[(4-isobutoxyphenyl)methyl]]-1,5-dihydro-2,4-benzodiazepin -3-one:

[0322] Prepared according to the method of Example 1-3.8 to obtain 47.3 mg (yield 18.6%) of a white solid. 1 H NMR (400 MHz, DMSO-d6) δ 1 H NMR (400 MHz, DMSO-d6) δ 8.53 (s, 1H), 7.23 - 7.17 (m, 2H), 7.09 (dd, J = 9.0, 2.6 Hz, 1H), 7.02 (dd, J = 8.3, 5.5 Hz, 1H), 6.94 (td, J = 8.6, 2.6 Hz, 1H), 6.89 - 6.82 (m, 2H), 4.67 - 4.28 (m, 7H), 3.73 (d, J = 6.4 Hz, 2H), 3.70 - 3.55 (m, 2H), 3.28 (dd, J = 11.9, 8.8 Hz, 1H), 3.04 (td, J = 10.5, 8.2 Hz, 1H), 2.87 (s, 3H), 2.55 - 2.43 (m, 1H), 2.15 (ddt, J = 13.6, 8.8, 4.6 Hz, 1H), 2.05 (dq, J = 13.3, 6.7 Hz, 1H), 1.04 (d, J = 6.7 Hz, 6H); LCMS (ES, m / z): 426 [M + H] + .

[0323] The synthetic general formulas of Compound A19014-0 and A19014-0A are shown as follows:

[0324]

[0325] Examples 1 - 13: Preparation of (3R,4S)-8-fluoro-2-(1-methyl-3-fluoropiperidin-4-yl)-4-[[4-(2-methylpropoxy)phenyl]methyl]-1,5-dihydro-2H-1,4-benzodiazepin-3-one (A19014-0): - Preparation of 4-amino-3-fluoropiperidine-1-carboxylic acid benzyl ester:

[0326]

[0327] 1 - 13.1 Preparation of 4-amino-3-fluoropiperidine-1-carboxylic acid benzyl ester:

[0328] 3-Fluoro-4-oxopiperidine-1-carboxylic acid benzyl ester (2 g, 7.9 mmol), acetamide (2.35 g, 39.8 mmol), NaBH3CN (1 g, 15.9 mmol), and methanol (10 mL) were placed in a 25 mL three-necked round-bottom flask. The flask was purged with nitrogen and the reaction was carried out at room temperature overnight. After completion of the reaction, the mixture was extracted with EtOAc (3 × 30 mL), and the resulting mixture was washed with brine (2 × 20 mL), dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography using an ethyl acetate / petroleum ether (1:1) system to give 0.7 g (yield 34.86%) of a colorless oil.

[0329] 1 - 13.2 Preparation of 4-[2-[[(tert-butoxycarbonyl)amino]methyl]-5-fluorobenzamido]-3-fluoropiperidine-1-carboxylic acid benzyl ester:

[0330] Prepared according to the method of Example 1 - 2.3 to obtain 1.3 g of an oil.

[0331] 1 - 13.3 Preparation of 4-[2-(aminomethyl)-5-fluorobenzamido]-3-fluoropiperidine-1-carboxylic acid benzyl ester:

[0332] Prepared according to the method of Example 1 - 1.10 to obtain 1 g of a light yellow oil.

[0333] 1 - 13.4 Preparation of 3-fluoro-4-[5-fluoro-2-[[[4-(2-methylpropoxy)phenyl]methyl]methylamino}benzamido]-piperidine-1-carboxylic acid benzyl ester:

[0334] Prepared according to the method of Example 1 - 2.5 to obtain 0.6 g (42.8%) of a yellow oil.

[0335] 1 - 13.5 Preparation of 3-fluoro-4-[5-fluoro-2-[[4-(2-methylpropoxy)benzyl]methylamino]benzamido]-piperidine-1-carboxylic acid benzyl ester:

[0336] Prepared according to the method of Example 1 - 2.6 to obtain 0.23 g (18.32%) of a yellow oil.

[0337] 1-13.6 8-Fluoro-2-(benzyl 3-fluoropiperidine-1-carboxylate)-4-[[4-(2-methylpropoxy)phenyl]methyl]-1,5-dihydro-2,4-benzodiazepin -3-one Preparation:

[0338] Prepared according to the method of Example 1-1.9 to obtain 0.13 g (yield 88.7%) of a colorless oil.

[0339] 1-13.7 (3R,4S)-8-Fluoro-2-(1-methyl-3-fluoropiperidin-4-yl)-4-[[4-(2-methylpropoxy)phenyl]methyl]-1,5-dihydro-2,4-benzodiazepin -3-one Preparation:

[0340] Prepared according to the experimental method of Example 1-3.8, and then 18.1 mg (yield 17.03%) of a yellow solid was obtained by Flash-Prep-HPLC. 1 H NMR (400 MHz, Methanol-d4) δ 7.23 - 7.18 (m, 2H), 7.13 (dd, J = 9.0, 2.6 Hz, 1H), 7.00 (dd, J = 8.4, 5.5 Hz, 1H), 6.93 (td, J = 8.6, 2.6 Hz, 1H), 6.88 - 6.84 (m, 2H), 4.85 - 4.77 (m, 1H), 4.71 (d, J = 5.1 Hz, 1H), 4.59 - 4.40 (m, 5H), 4.35 - 4.21 (m, 2H), 3.74 (d, J = 6.5 Hz, 2H), 3.26 (dt, J = 10.5, 5.3 Hz, 0H), 2.98 2.84 (m, 1H), 2.38 (s, 3H), 2.22 1.90 (m, 4H), 1.87 1.71 (m, 1H), 1.04 (d, J = 6.7 Hz, 6H); LCMS (ES, m / z): 458 [M+H] + .

[0341] Example 1-14: (3S,4S)-8-Fluoro-2-(1-methyl-3-fluoropiperidin-4-yl)-4-[[4-(2-methylpropoxy)phenyl]methyl]-1,5-dihydro-2,4-benzodiazepin -3-one Preparation (A19014-0A):

[0342]

[0343] Prepared according to the experimental method in Example 1-13 to obtain 84 mg (yield 74.25%) of a colorless oil. 11H NMR (400 MHz, Methanol-d4) δ 7.22 (d, J = 8.5 Hz, 2H), 7.07 (dd, J = 9.1, 2.7 Hz, 1H), 7.01 (dd, J = 8.4, 5.5 Hz, 1H), 6.93 (td, J = 8.6, 2.7 Hz, 1H), 6.89–6.84 (m, 2H), 4.80 - 4.19 (m, 8H), 3.74 (d, J = 6.5 Hz, 2H), 3.24 - 3.11 (m, 1H), 3.03 (d, J = 11.5 Hz, 1H), 2.49 - 2.17 (m, 6H), 2.06 (dt, J = 13.3, 6.6 Hz, 1H), 1.60 (d, J = 12.4 Hz, 1H), 1.09 - 0.99 (m, 6H); LCMS (ES, m / z): 458 [M+H] + .

[0344] Examples 1 - 15: Preparation of 7,8-difluoro-2-(4-isobutoxybenzyl)-4-(1-methylpiperidin-4-yl)-1,2,4,5-tetrahydro-3H-benzo[e][1,3]diazepin -3-one (A20001):

[0345]

[0346] Prepared according to the experimental method in Example 1 - 1 to obtain 51.9 mg, white solid. 1 1H NMR (400 MHz, Methanol-d4) δ 7.34 (dd, J = 10.0, 8.4 Hz, 1H), 7.20 (d, J = 8.4 Hz, 2H), 6.92 (dd, J = 10.0, 8.4 Hz, 1H), 6.90–6.82 (m, 2H), 4.47 (d, J = 14.0 Hz, 4H), 4.36 (s, 2H), 4.40–4.28 (m, 1H), 3.73 (d, J = 6.5 Hz, 2H), 3.60 (d, J = 12.2 Hz, 2H), 3.21–3.11 (m, 2H), 2.90 (s, 3H), 2.35 (dd, J = 13.5, 3.8 Hz, 1H), 2.29 (dd, J = 13.1, 3.8 Hz, 1H), 2.05 (hept, J = 6.7 Hz, 1H), 1.94 (d, J = 13.6 Hz, 2H), 1.04 (d, J = 6.7 Hz, 6H). LCMS (ES, m / z): 458 [M+H] + 。

[0347] Examples 1 - 16: Preparation of 7 - fluoro - 2 - [[4 - (2 - hydroxy - 2 - methylpropyl)phenyl]methyl] - 4 - (1 - methylpiperidin - 4 - yl) - 1,5 - dihydro - 2,4 - benzodiazepin -3 - one (A19010):

[0348]

[0349] Synthetic route:

[0350]

[0351] 1 - 16.1 Preparation of benzyl 4 - [4 - [(4 - bromophenyl)methyl] - 8 - fluoro - 3 - oxo - 1,5 - dihydro - 2,4 - benzodiazepin -2 - yl]piperidine - 1 - carboxylate:

[0352] Prepared according to the method in Examples 1 - 4, and 1 g of light yellow oil was obtained.

[0353] 1 - 16.2 Preparation of benzyl 4 - [4 - ([4 - [(E) - 2 - ethoxyvinyl]phenyl]methyl) - 8 - fluoro - 3 - oxo - 1,5 - dihydro - 2,4 - benzodiazepin -2 - yl]piperidine - 1 - carboxylate:

[0354] Add benzyl 4 - [4 - [(4 - bromophenyl)methyl] - 8 - fluoro - 3 - oxo - 1,5 - dihydro - 2,4 - benzodiazepin -2 - yl]piperidine - 1 - carboxylate (1.00 g, 1.765 mmol), (E) - 1 - ethoxyvinyl - 2 - boronic acid pinacol ester (0.70 g, 3.531 mmol), dioxane (10.00 mL), water (2.00 mg) and K3PO4 (1.12 g, 5.296 mmol) into a 100 - ml three - necked flask. Stir the resulting solution at 25 °C for 10 minutes, add Pd(dppf)Cl2 (0.14 g, 0.177 mmol) to it, and heat up to 100 °C for reaction for 2 hours. After the reaction is completed, add water to quench the reaction, extract with ethyl acetate (3x20 mL), wash the organic phase with 1N hydrochloric acid solution (2x20 mL), then wash with saturated brine (2x20 ml), dry the organic phase with anhydrous sodium sulfate, evaporate the solvent under reduced pressure, and purify by column chromatography (EA:PE = 1:30) to obtain 400 mg of the compound benzyl 4 - [4 - ([4 - [(E) - 2 - ethoxyvinyl]phenyl]methyl) - 8 - fluoro - 3 - oxo - 1,5 - dihydro - 2,4 - benzodiazepin -2 - yl]piperidine - 1 - carboxylate, with a yield of 40.63%, as a brown oily substance.

[0355] 1-16.3 Preparation of benzyl 4-(8-fluoro-3-oxo-4-[[4-(2-oxoethyl)phenyl]methyl]-1,5-dihydro-2,4-benzodiazepin -2-yl)piperidine-1-carboxylate:

[0356] To a 50 mL three-necked flask were added benzyl 4-[4-([4-[(E)-2-ethoxyvinyl]phenyl]methyl)-8-fluoro-3-oxo-1,5-dihydro-2,4-benzodiazepin -2-yl]piperidine-1-carboxylate (200.00 mg, 0.359 mmol), tetrahydrofuran (2.00 mL) and HCl (6 M) (2.00 mL), and the mixture was stirred at 25 °C for 2 hours. After the reaction was completed, the reaction was quenched with water, extracted with methyl tert-butyl ether (3 x 10 mL), the organic phase was washed with 30 mL of saturated brine, dried over anhydrous sodium sulfate, and the solvent was removed under reduced pressure to obtain 200 mg of benzyl 4-(8-fluoro-3-oxo-4-[[4-(2-oxoethyl)phenyl]methyl]-1,5-dihydro-2,4-benzodiazepin -2-yl]piperidine-1-carboxylate, with a yield of 105.30%, as a yellow oil.

[0357] 1-16.4 Preparation of benzyl 4-[8-fluoro-4-[[4-(2-hydroxypropyl)phenyl]methyl]-3-oxo-1,5-dihydro-2,4-benzodiazepin -2-yl]piperidine-1-carboxylate:

[0358] To a 50 mL three-necked flask were added benzyl 4-(8-fluoro-3-oxo-4-[[4-(2-oxoethyl)phenyl]methyl]-1,5-dihydro-2,4-benzodiazepin -2-yl)piperidine-1-carboxylate (200.00 mg, 0.378 mmol) and tetrahydrofuran (3.00 mL), the temperature was lowered to 0 °C and stirred for 5 minutes, and methylmagnesium bromide (180.12 mg, 1.511 mmol) was added thereto, and the temperature was raised to 60 °C and reacted for 2 hours. After the reaction was completed, the reaction was quenched with NH4Cl solution, extracted with ethyl acetate (3 x 20 mL), the organic phase was washed with 20 ml of saturated brine, dried over anhydrous sodium sulfate, and the solvent was removed under reduced pressure to obtain 200 mg of benzyl 4-[8-fluoro-4-[[4-(2-hydroxypropyl)phenyl]methyl]-3-oxo-1,5-dihydro-2,4-benzodiazepin -2-yl]piperidine-1-carboxylate, with a yield of 97.06%, as a yellow oil.

[0359] 1-16.5 Benzyl 4-[8-fluoro-3-oxo-4-[[4-(2-oxopropyl)phenyl]methyl]-1,5-dihydro-2,4-benzodiazepin Preparation of 2-amino-2-piperidin-1-carboxylate:

[0360] In a 50 mL three-necked flask, benzyl-4-[8-fluoro-4-[[4-(2-hydroxypropyl)phenyl]methyl]-3-oxo-1,5-dihydro-2,4-benzodiazepine -2-yl]piperidine-1-carboxylate (200.00 mg, 0.367 mmol) and dichloromethane (20.00 mL), nitrogen protection, cooling to 0 ° C and stirring for 5 minutes, then adding DMP (310.92 mg, 0.733 mmol), stirring at 25 ° C for 3 hours. After the reaction is completed, NaHCO3 solution is added to quench the reaction, and it is extracted with dichloromethane (3x20 mL), the organic phase is washed with saturated brine, dried over anhydrous sodium sulfate, and the solvent is evaporated under reduced pressure to obtain the compound benzyl 4-[8-fluoro-3-oxy-4-[[4-(2-oxypropyl)phenyl]methyl]-1,5-dihydro-2,4-benzodiazepine -2-yl]piperidine-1-carboxylate 180 mg, yield 90.33%, yellow oil.

[0361] 1-16.6Benzyl-4-[8-fluoro-4-[[4-(2-hydroxy-2-methylpropyl)phenyl]methyl]-3-oxo-1,5-dihydro-2,4-benzodiazepine Preparation of 2-amino-2-piperidin-1-carboxylate:

[0362] In a 50 mL three-necked flask, benzyl-4-[8-fluoro-3-oxo-4-[[4-(2-oxopropyl)phenyl]methyl]-1,5-dihydro-2,4-benzodiazepine -2-yl]piperidine-1-carboxylate (150.00 mg, 0.276 mmol), tetrahydrofuran (5.00 mL, 0.069 mmol) and MeMgBr (2.00 mL, 0.017 mmol), nitrogen protection, heated to 60 ° C for 3 hours. After the reaction was completed, the reaction was quenched with NH4Cl solution, extracted with ethyl acetate (3x10 mL), and the organic phase was washed with 10 ml of saturated brine, dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure to obtain the compound benzyl-4-(8-fluoro-4-[[4-(2-hydroxy-2-methylpropyl)phenyl]methyl]-3-oxo-1,5-dihydro-2,4-benzodiazepine -2-yl)piperidine-1-carboxylate 140 mg, yield 90.66%, yellow oil.

[0363] 1-16.7 7-Fluoro-2-[[4-(2-hydroxy-2-methylpropyl)phenyl]methyl]-4-(1-methylpiperidin-4-yl)-1,5-dihydro-2,4-benzodiazepine Preparation of -3-ketone:

[0364] To a 100 mL three-necked flask, add benzyl-4-[8-fluoro-4-[[4-(2-hydroxy-2-methylpropyl)phenyl]methyl]-3-oxo-1,5-dihydro-2,4-benzodiazepin -2-yl]piperidine-1-carboxylate (150.00 mg, 0.268 mmol), methanol (10.00 mL, 246.989 mmol), formaldehyde (2.00 mL, 0.067 mmol) and Pd(OH)2 / C (20.00 mg, 0.142 mmol). Introduce hydrogen gas into the reaction system, stir at 25 °C for 5 hours, quench the reaction by adding water, extract with ethyl acetate (3 x 10 mL), wash the organic phase with 10 mL of saturated brine, dry over anhydrous sodium sulfate, evaporate the solvent under reduced pressure, and purify by column chromatography (DCM:MeOH = 30:1) to obtain 45 mg of compound 7-fluoro-2-[[4-(2-hydroxy-2-methylpropyl)phenyl]methyl]-4-(1-methylpiperidin-4-yl)-1,5-dihydro-2,4-benzodiazepin -3-one, with a yield of 38.19%, a yellow oil. 1 H NMR (400 MHz, Methanol-d4) δ 8.48 (s, 1H, FA), 7.10–6.98 (m, 4H), 6.93 (td, J = 8.6, 2.7 Hz, 1H), 6.58 (d, 2H), 4.42 (s, 4H), 4.36 (s, 2H), 4.30–4.18 (m, 1H), 3.46–3.39 (m, 2H), 2.96–2.83 (m, 4H), 2.77 (s, 3H), 2.25–2.05 (m, 2H), 1.96–1.84 (m, 3H), 1.36–1.30 (m, 1H), 0.99 (d, J = 6.6 Hz, 6H). LCMS (ES, m / z): 400 [M+H] + 。

[0365] Examples 1-17: Preparation of 4-[(4-cyclopropoxyphenyl)methyl]-7-fluoro-2-(1-methylpiperidin-4-yl)-1,5-dihydro-2,4-benzodiazepin -3-one (A19022):

[0366]

[0367] Prepared 9.5 mg of white solid by referring to the experimental method in Examples 1-2. 1H-NMR (400 MHz, Chloroform-d): δ 7.25 (d, J = 8.5 Hz, 2H), 7.15 (t, J = 7.0 Hz, 1H), 7.04–6.97 (m, 2H), 6.90 (td, J = 8.4, 2.6 Hz, 1H), 6.65 (dd, J = 9.1, 2.6 Hz, 1H), 4.45 (s, 2H), 4.40 (s, 2H), 4.25 (s, 3H), 3.74 (p, J = 4.5 Hz, 1H), 3.03 (s, 2H), 2.42–2.38 (m, 3H), 2.22 (s, 2H), 1.99 (s, 2H), 1.73 (d, J = 12.5 Hz, 2H), 0.78 (d, J = 4.5 Hz, 4H). LCMS (ES, m / z): 424 [M+1] + 。

[0368] Example 2. Preparation of the salt of the compound represented by General Formula I

[0369] Dissolve the free base of the compound represented by General Formula I in an organic solvent to form a solution. Optionally dissolve the acid in an organic solvent. Drop the organic solvent - acid solution into the solution of the free base of the compound represented by General Formula I, stir, separate and dry to obtain the salt of the compound represented by General Formula I; the organic solvents include but are not limited to absolute ethanol, ethyl acetate, etc.; the acids include but are not limited to phosphoric acid, citric acid, etc.

[0370] Only A19006 is taken as an example below. Those skilled in the art should understand that the other example compounds above can also be prepared, characterized and confirmed with reference to Example 2.

[0371] Example 2-1. Preparation of A19006 phosphate

[0372] Preparation method: Weigh about 5.2 g of the A19006 base sample into a 250 mL single-necked flask, add 100 ml of ethyl acetate, and stir until completely dissolved; weigh 1.4 g of phosphoric acid (85%) and dissolve it in 20 mL of ethyl acetate, and slowly drop the ethyl acetate - phosphoric acid solution under stirring; after 1 h, the dropping is completed, continue to stir at room temperature for 1 h, and stop the reaction; perform suction filtration, wash the filter cake with ethyl acetate (2×20 ml), and dry it in vacuo at 45 °C for 3 hours to obtain 6.0 g of A19006 phosphate, yield: 93.7%, 11H NMR (400 MHz, DMSO-d6) δ 7.33 (dd, J = 9.3, 2.7 Hz, 1H), 7.23 (d, J = 8.3 Hz, 2H), 7.10 (dd, J = 8.4, 5.7 Hz, 1H), 7.04–6.94 (m, 3H), 4.44 (s, 2H), 4.38 (s, 2H), 4.30 (s, 2H), 4.19–4.28 (m, 1H), 3.79 (tt, J = 6.1, 3.0 Hz, 1H), 3.23 (d, J = 11.5 Hz, 2H), 2.85–2.64 (m, 2H), 2.56 (s, 3H), 2.26 (qd, J = 13.1, 4.1 Hz, 2H), 1.58 (dd, J = 13.1, 3.8 Hz, 2H), 0.80–0.72 (m, 2H), 0.63 (q, J = 3.4, 2.9 Hz, 2H).

[0373] Example 2-2. Preparation of A19006 Citrate

[0374] Preparation method: Weigh about 10 g of A19006 base sample into a 500 mL single-necked flask, add 200 ml of absolute ethanol, and stir until completely dissolved; weigh 5.0 g of anhydrous citric acid and dissolve it in 40 mL of absolute ethanol. Slowly add the absolute ethanol-citric acid solution dropwise under stirring. A large amount of white solid will suddenly precipitate during the dropping process; after 1 h, the dropping is completed. Transfer the flask to an ice-water bath for cooling and keep stirring to precipitate the solid; perform suction filtration, wash the filter cake with absolute ethanol (2 × 40 ml), and dry it under vacuum at 45 °C for 3 hours to obtain A19006 citrate. Yield: 12.3 g; Yield rate: 84.6%. 1 1H NMR (400 MHz, DMSO-d6) δ 7.24 (d, J = 8.4 Hz, 2H), 7.11–7.21 (m, 2H), 7.00–7.07 (m, 1H), 6.98 (d, J = 8.3 Hz, 2H), 4.38 (s, 4H), 4.32 (s, 2H), 4.18 (tt, J = 12.2, 4.0 Hz, 1H), 3.80 (tt, J = 6.0, 2.9 Hz, 1H), 3.31 (d, J = 11.8 Hz, 2H), 2.93–2.78 (m, 2H), 2.66 (d, J = 2.8 Hz, 3H), 2.09 (qd, J = 13.0, 4.0 Hz, 2H), 1.66 (dd, J = 13.7, 3.8 Hz, 2H), 0.81–0.73 (m, 2H), 0.63 (q, J = 3.4, 3.0 Hz, 2H).

[0375] Take the obtained A19006 citrate above. After testing, its X-ray powder diffraction pattern is shown in Figure 1 and its DSC spectrum is shown inFigure 2 During the DSC heating process, the starting point of the endothermic peak is 164.7 °C, the ending point is 170.0 °C, and the peak value is 167.7 °C. This crystal form is defined as crystal form I, and the characteristic peak positions are shown in Table 1 below:

[0376] Table 1 X-ray powder diffraction

[0377]

[0378]

[0379] Test example

[0380] Test Example 1 Hygroscopicity test of salt form

[0381] Refer to the guiding principle for the hygroscopicity test of drugs in Part IV of the Chinese Pharmacopoeia 2020, item 9103.

[0382] Experimental method:

[0383] (1) Take a dry stoppered glass weighing bottle (outer diameter 50 mm, height 15 mm), and place it in a suitable constant temperature dryer (with ammonium chloride or ammonium sulfate saturated solution placed at the bottom) at 25 °C ± 1 °C or an artificial climate chamber (set temperature 25 °C ± 1 °C, relative humidity 80% ± 2%) one day before the test, and accurately weigh its weight (m1).

[0384] (2) Take an appropriate amount of the test substance and spread it evenly in the above-mentioned weighing bottle. The thickness of the test substance is generally about 1 mm, and accurately weigh its weight (m2).

[0385] (3) Open the weighing bottle and place it together with the bottle cap under the above constant temperature and humidity conditions for 24 hours.

[0386] (4) Cover the weighing bottle cap and accurately weigh its weight (m3).

[0387]

[0388] Experimental results: The specific experimental results are shown in Table 2.

[0389] Table 2 Hygroscopicity determination results of salt form A19006

[0390]

[0391] Conclusion: As can be seen from the above table, A19006 phosphate has low hygroscopicity.

[0392] Test Example 2 Solubility test

[0393] Refer to the general rules in Part IV of the Chinese Pharmacopoeia 2020.

[0394] Experimental method: Weigh the test sample ground into fine powder or measure the liquid test sample, place it in a solvent of a certain volume at 25°C ± 2°C, shake it strongly for 30 seconds every 5 minutes, observe the dissolution situation within 30 minutes, and if there are no visually visible solute particles or droplets, it is regarded as completely dissolved.

[0395] Experimental results: The specific experimental results are shown in Table 3.

[0396] Table 3 Solubility of A19006 salt form

[0397]

[0398] Conclusion: As can be seen from the above table, A19006 citrate dissolves in the pH 1 solution and is slightly soluble in the pH 7 solution.

[0399] Test Example 3 Stability test

[0400] Experimental method:

[0401] (1) High humidity experiment

[0402] Place the A19006 citrate polymorph I sample in a clean watch glass, place it at 25°C under a relative humidity of 92.5% or at room temperature under a relative humidity of 75% ± 1% for 30 days, and take samples on the 0th day and the 30th day. Conduct XRPD, DSC, and HPLC analyses.

[0403] (2) Light exposure experiment

[0404] Place the A19006 citrate polymorph I sample in a clean watch glass, lay it flat and place it in a light box (illuminance of 5000 lx ± 500 lx, ultraviolet light intensity of 90 μW / cm2) for 30 days, and take samples on the 0th day and the 30th day. Conduct XRPD, DSC, and HPLC analyses.

[0405] Experimental results: The specific experimental results are shown in Table 4.

[0406] Table 4 HPLC determination results of the stability of A19006 citrate polymorph I

[0407]

[0408] Remark: Main peak content (%) Area normalization method

[0409] Conclusion: As can be seen from the above table, A19006 citrate polymorph I has good stability under high humidity and light exposure.

[0410] Those skilled in the art can also expect that the stability, hygroscopicity, etc. of the citrate salts of other compounds are also better.

[0411] Test Example 4 In vitro receptor binding assay

[0412] Experimental methods

[0413] 4.1 Preparation of solutions required for experiments

[0414] A: (for preparing 5-HT 2C receptor membrane): 50 mM Tris-HCl buffer: Dissolve 96.8 g of Tris in double-distilled water to a total volume of 4000 ml, adjust the pH to 7.5 with HCl, dilute to 16000 mL, pH = 7.4

[0415] B: (for preparing 5-HT 2A receptor membrane): Weigh 11.7 mg of EDTA and 380.84 mg of MgCl2, add to 50 mM Tris-HCl buffer with a total volume of 400 mL, adjust the pH = 7.4. Make the final concentrations of EDTA 0.1 mM and MgCl2 10 mM respectively.

[0416] C: (for preparing Dopamine receptor membrane): Weigh 2.978 g of HEPES, 1.17 g of NaCl, 0.119 g of MgCl2, and 36.5 mg of EDTA, add to pure water with a total volume of 250 ml, adjust the pH = 7.4. Make the final concentrations 50 mM HEPES, 50 mM NaCl, 5 mM MgCl2, 0.5 mM EDTA, pH 7.4 respectively.

[0417] 4.2 Preparation of receptor membranes

[0418] 1) Preparation of CHO-5-HT 2A receptor membrane

[0419] CHO-5-HT 2A Cells are taken out from the -80 °C refrigerator and thawed naturally, centrifuged at 2000 g and 4 °C for 15 minutes. Take the precipitate and discard the supernatant. Add solution B to the precipitate. Mix the cells for 20 - 30 seconds, then centrifuge at 50000 g and 4 °C for 25 min. Carefully discard the upper layer of liquid, add solution B again and mix well, then centrifuge at 50000 g and 4 °C for 25 min. Store the precipitate at -80 °C.

[0420] 2) Preparation of 5-HT 2C membrane

[0421] The rat cortex is taken out from the -80 °C refrigerator and thawed naturally, added to solution A and homogenized at speed 4 for 3 - 4 s, homogenized 4 times, centrifuged at 50000 g and 4 °C for 25 min, discard the supernatant, add solution A, mix well with a vortex mixer, centrifuge at 50000 g and 4 °C for 25 min, repeat the centrifugation twice. After centrifugation, discard the supernatant, and store the precipitate at -80 °C for later use.

[0422] 3) Preparation of CHO-D2 receptor membrane

[0423] After the CHO-D2 cells were taken out of the -80 °C refrigerator and thawed naturally, they were centrifuged at 2000 g for 15 min. The precipitate was added with homogenate C, mixed well with a vortex mixer, centrifuged at 50000 g at 4 °C for 25 min, the supernatant was discarded, the precipitate was taken, and C buffer was added again for washing, resuspension and centrifugation. After centrifugation, the supernatant was discarded, and the precipitate was stored at -80 °C for standby.

[0424] 4.3 Receptor competition binding experiment

[0425] 1) 5-HT 2A Receptor competition binding assay

[0426] Step 1: First, the prepared membrane was made into a 10 mg / mL membrane suspension with homogenate B for standby.

[0427] Step 2: 100 μL of the membrane preparation was added to each reaction tube.

[0428] Step 3: 100 μL of solution B was added to the total binding tube (TB), 100 μL of Methysergide (final concentration 1.0×10 -5 M) was added to the non-specific binding tube (NB), and 100 μL of the test compound was added to each test compound tube (CB).

[0429] Step 4: 10 μL of radioactive ligand 3 3H-Ketanserin was added to each reaction tube, with a final concentration of 2.98 nM.

[0430] Step 5: The reaction tubes were incubated at 37 °C for 25 min. After the reaction was completed, the bound ligand was quickly filtered under reduced pressure through a Whatman GF / C filter paper (pre-soaked with 0.5% PEI for more than 1 h). After filtration, the filter membrane was dried at 60 °C, the bottom membrane was attached, 40 μL of scintillation fluid was added, the top membrane was sealed, and it was left standing.

[0431] Step 6: The scintillation vial was placed in a liquid scintillation counter for counting.

[0432] 2) 5-HT 2C Receptor competition binding assay

[0433] Step 1: First, the prepared membrane was made into a 210 mg / mL membrane suspension with homogenate B for standby.

[0434] Step 2: 100 μL of the membrane preparation was added to each reaction tube.

[0435] Step 3: 100 μL of solution B was added to the total binding tube (TB), Ketanserin (final concentration 1.0×10-5 100 μL of M), and 100 μL of the test compound was added to each test compound tube (CB).

[0436] Step 4: Add the radioactive ligand to each reaction tube 3 10 μL of ³H-Mesulergine, final concentration 3 nM.

[0437] Step 5: Incubate each reaction tube at 37 °C for 25 min. After the reaction is complete, the bound ligand is rapidly filtered under reduced pressure. Whatman filter paper GF / C was saturated with 0.5% PEI solution 1 h in advance, washed thoroughly with ice-cold Tris buffer, the filter was taken out and placed in a 4 mL scintillation vial, and 1 mL of toluene scintillation fluid was added and mixed well.

[0438] Step 6: Place the scintillation vial into a liquid scintillation counter for counting.

[0439] 3) CHO-D2 receptor competitive binding assay

[0440] Step 1: First, prepare the membrane with homogenate C to make a suspension of 8 mg / mL membrane for standby.

[0441] Step 2: Add 100 μL of the membrane preparation to each reaction tube.

[0442] Step 3: Add 100 μL of solution C to the total binding tube (TB), 100 μL of Haloperidol (final concentration 1.0×10⁻⁶ M) to the non-specific binding tube (NB), and 100 μL of the test compound to each test compound binding tube (CB). -5 100 μL of the test compound was added to each test compound binding tube (CB).

[0443] Step 4: Add the radioactive ligand to each reaction tube 3 10 μL of ³H-Spiperone, final concentration 1.176 nM.

[0444] Step 5: Incubate each reaction tube at 37 °C for 25 min. After the reaction is complete, the bound ligand is rapidly filtered under reduced pressure. Whatman filter paper GF / B plate was soaked with 0.5% PEI for more than 1 h in advance. After filtration, the filter membrane was dried at 60 °C, a bottom film was attached, 40 μL of scintillation fluid was added, the top film was sealed, and left standing.

[0445] Step 6: Place the suction filter plate into a liquid scintillation counter for counting.

[0446] 4.4. Experimental results

[0447] Pimaserine 5-HT 2A 、5-HT 2C The Ki values of the receptor were 0.036 and 2.94 nM respectively. For the 5-HT of compound NH-K-A19016-OA 2A, 5-HT 2C The Ki values for the receptors are 0.002 and 26.10 nM respectively, superior to pimaserine; for compound NH-K-A19001, the Ki values for 5-HT 2A , 5-HT 2C receptors are 0.028 and 2.40 nM respectively, superior to pimaserine; for compound NH-K-A19005 and 5-HT 2A , 5-HT 2C receptors, the Ki values are 0.43 and 3.39 nM respectively, at the same level as pimaserine 862. See the following table, Table 5.

[0448] Table 5 In vitro receptor binding

[0449]

[0450]

[0451] Test Example 5: In vitro hERG experiment

[0452] 5.1. Compound preparation

[0453] a. Sequentially dilute the stock solution of the test compound with DMSO to dilution solutions of 0.3 mM, 1 mM, and 3 mM.

[0454] b. Dilute the stock solution of the test compound with extracellular fluid to obtain working solutions of the test compound at concentrations of 0.3 μΜ, 1 μΜ, 3 μM, 10 μΜ, and 30 μΜ. Ultrasonicate all the working solutions of the test compound for 20 min.

[0455] c. Dissolve 10 mg of cisapride (the positive compound in the system) in 2002.42 μL of dimethyl sulfoxide (DMSO) to prepare a stock solution of 10.113 mM.

[0456] d. Sequentially dilute the cisapride stock solution with dimethyl sulfoxide (DMSO) to 1 μM, 10 μM, 100 μM, and 1 mM.

[0457] e. Take 10 μL of each concentration and add it to 10 mL of extracellular fluid to ensure that the DMSO concentration is 0.1%.

[0458] f. The final working solution concentration of cisapride is 1 nM, 10 nM, 100 nM, and 1000 nM.

[0459] g. For all the test working solution concentrations, visually observed, there is no visible precipitation.

[0460] 5.2. Cell line information

[0461] In this experiment, we used the HEK-293 cell line stably expressing the hERG potassium channel for experimental detection.

[0462] The HEK-293 cell line stably expressing the hERG potassium channel was cultured in DMEM medium containing 10% fetal bovine serum and 0.8 mg / mL G418 at a culture temperature of 37 °C and a carbon dioxide concentration of 5%.

[0463] Cell passage: Remove the old medium and wash once with PBS, then add 0.5 mL of TrypLE TM Express solution and incubate at 37 °C for 1 minute. When the cells detach from the bottom of the dish, add 3 mL of pre-warmed complete medium at 37 °C. Gently pipette the cell suspension to separate the aggregated cells. Transfer the cell suspension to a sterile centrifuge tube and centrifuge at 300 G for 5 minutes to collect the cells. Inoculate the cells into 6-cm cell culture dishes, with an inoculation amount of 1×10 5 cells per cell culture dish (final volume: 5 mL) for amplification or maintenance culture.

[0464] For patch clamp detection, seed 5×10 3 cells onto cover slips and culture in 24-well plates (final volume: 500 μL). After 18 hours, perform experimental detection.

[0465] To maintain the electrophysiological activity of the cells, the cell density must not exceed 80%.

[0466] 5.3. Patch Clamp Detection

[0467] Under an inverted microscope, use a glass electrode micromanipulator (micromanipulator) to bring the recording electrode into contact with the cell, and then apply negative pressure to promote the formation of a GΩ seal. After the formation of a GΩ seal, perform rapid capacitance compensation, and then continuously apply negative pressure to rupture the cell membrane to form a whole-cell recording mode. In the whole-cell recording mode, perform slow capacitance compensation and record the values of membrane capacitance and series resistance.

[0468] The voltage stimulation protocol for the hERG potassium current in the cells is as follows: The cell membrane is clamped at -80 mV, then depolarized from -80 mV to +30 mV and maintained for 2.5 seconds, and then quickly held at -50 mV for 4 seconds to evoke the tail current of the hERG channel. Collect data every 10 seconds. Use -50 mV for leak current detection. See Figure 3 .

[0469] The cover glass with seeded cells was placed in the recording chamber of an inverted microscope. The negative control and the test compound were perfused from low to high concentrations by gravity perfusion through the recording chamber in sequence to act on the cells rapidly. During the recording, a vacuum pump was used for continuous circulation of the external solution. The current detected in each cell's negative control was used as the control group for the cell itself. Each drug concentration was allowed to act for 5 minutes or until the current stabilized. All experiments were conducted at room temperature.

[0470] 5.4. Data Analysis

[0471] First, standardize the current after the action of each drug concentration Then calculate the corresponding inhibition rate Calculate basic statistics for each concentration, including the mean (Mean), standard deviation (SD), standard error (SE), and the number of replicates (n). Fit the dose-dependent curve with the following equation and calculate the half-inhibitory concentration (IC 50 )

[0472]

[0473] where C represents the concentration of the test compound, IC 50 represents the half-inhibitory concentration, and h represents the Hill coefficient. Curve fitting and the calculation of IC 50 were completed using GraphPad Prism 5.0 software.

[0474] 5.5. Experimental Results

[0475] The IC50 of pimavanserin in the hERG experiment was 208 nM, and the IC50s of NH-K-A19001, NH-K-A19005, and NH-K-A19006 in the hERG experiment were 206, 3173, and 1194 nM respectively. The cardiotoxicity of these three compounds was less than that of pimavanserin, indicating that the compounds of the present invention have lower cardiotoxicity compared to pimavanserin. The results are shown in the following table.

[0476] Table 6

[0477] Compound hERG (nM) NH-K-A19001 260 NH-K-A19005 3173 NH-K-A19006 1194 Pimavanserin 208

[0478] Test Example 6: Animal Experiment

[0479] 6.1. Test Method

[0480] 6.1.1 Experiment on the effect on the MPTP + MK-801 Parkinson's psychosis mouse model (anti-PDP pharmacodynamic model)

[0481] Animals were intraperitoneally injected with different doses of MPTP every morning for 5 consecutive days. On the morning of the 5th day, after injecting MPTP, 1.5 h later, pimavanserin or NS was intraperitoneally injected. Then, 0.5 h later, MK-801 at a dose of 0.3 mg / kg (or NS) was intraperitoneally injected. After another 0.25 h, the mice were placed in an open-field activity box (a black polyethylene box with dimensions of 29 cm × 29 cm × 30 cm) for video recording for 20 min. After the video recording ended, video analysis was performed to evaluate the activity of the mice.

[0482] 6.1.2 Effects on the climbing behavior of male mice induced by MPTP + APO (DA motor deterioration model)

[0483] Animals were intraperitoneally injected with different doses of MPTP every morning for 5 consecutive days. On the morning of the 5th day, after injecting MPTP, 1.5 h later, pimavanserin, clozapine, quetiapine or NS was intraperitoneally injected. Then, 0.5 h later, 1 mg / kg of APO was subcutaneously injected (the administration volume was 0.1 ml / 10 g body weight). Immediately after subcutaneous injection, the mice were placed in a climbing cage (self-made by the company, a cylindrical cage with a diameter of 13 cm and a height of 15 cm, made of stainless steel wire mesh with a diameter of about 0.1 cm, the bottom is a semi-transparent polyethylene board, and the cage cover is a stainless steel cage cover). The behavior was observed and scored at the 10-11th, 20-21st, and 30-31st minutes after injecting APO.

[0484] The scoring criteria were as follows: 0 points were given when all four feet were on the floor; 1 point was given when the two front feet were on the wire cage; 2 points were given when all four feet were on the wire cage.

[0485] 6.1.3 Experiment on sedative side effects investigation (anti-sedation model)

[0486] Qualified SPF-grade C57BL / 6j mice were randomly divided into 13 groups, with 8 mice in each group: blank group, pimavanserin, NH-K-A1900, NH-K-A19005, NH-K-A19006. According to the different doses of each group, different concentrations of solutions were prepared for intraperitoneal injection, and the final administration volume was 10 ml / kg.

[0487] Forty-five minutes after administering compounds such as pimavanserin, open-field activity detection was performed on all groups, and the movement conditions from 0 to 20 min were video-recorded. The movement distance in 20 min was analyzed using Top Scan 3.00 software. The inhibition rate of each group after administration relative to the blank group was calculated, and combined with the statistical conclusions, the sedative effects of the compounds were comprehensively evaluated.

[0488] 6.1.4 Test results

[0489] It can be seen from the experiment that the PDP pharmacodynamic effect, sedation, and motor deterioration ED of pimavanserin 50They were 0.37 mg / kg, 6.79 mg / kg, and >30 mg / kg respectively, and their sedation / PDP pharmacodynamic ratios were 18.35, and the movement deterioration / PDP pharmacodynamic ratios were >81.08; the PDP pharmacodynamics and sedation ED of NH-K-A19001 50 They were 0.33 mg / kg and 3.74 mg / kg respectively, and their sedation / PDP pharmacodynamic ratios were 11.33. The PDP pharmacodynamics of NH-K-A19005 was 1.85 mg / kg. The PDP pharmacodynamics and sedation ED of NH-K-A19006 50 They were 0.31 mg / kg and 11.9 mg / kg respectively. It can be seen that the pharmacodynamics of NH-K-A19001 and NH-K-A19006 were comparable to those of pimavanserin. The PDP pharmacodynamic dose of NH-K-A19005 was slightly higher than that of pimavanserin. In addition, the compounds of the present invention (such as NH-K-A19001, NH-K-A19005, and NH-K-A19016) did not have a DA mechanism of action, and no movement deterioration was observed.

[0490] Table 7

[0491]

[0492] Test Example 7: In vitro and in vivo experimental methods and data

[0493] 7.1. Mouse head-tossing test

[0494] 7.1.1 Test method

[0495] After mice were stratified by body weight, they were randomly divided into a model control group, a blank control group, and each dosing group. 1 h after the animals were given the vehicle or the drug by gavage, the animals were placed in a beaker (13 cm in diameter and 19 cm in height) lined with fresh bedding, and the modeling drug DOI ((±)-1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane hydrochloride, (±)-2,5-dimethoxy-4-iodoamphetamine hydrochloride) was injected intraperitoneally at a dose of 1 mg / kg, and the number of head tosses of the mice within 0 - 20 minutes after the intraperitoneal injection of DOI was recorded. The head-tossing behavior was defined as the rapid rotational twitching or wet dog-like shaking of the mouse's head, and this action should be distinguished from normal grooming or exploratory behaviors.

[0496] 7.1.2 Test data

[0497] The results of this test showed that the ED of pimavanserin in inhibiting DOI-induced head-tossing behavior in mice 50 was 0.39 mg / kg, and the ED of NH-K-A19001, NH-K-A19005, NH-K-A19006, and NH-K-A19012 in inhibiting DOI-induced head-tossing behavior in mice 50They were 0.06, 0.15, 0.012, and 0.30 mg / kg respectively, indicating that the compounds of the present invention have better effects against mental diseases and better drug efficacy. The detailed results are shown in the following table.

[0498] Table 8 ED of Compounds Such as Pimaserine in Inhibiting DOI-Induced Head-Tossing Behavior in Mice 50

[0499]

[0500]

[0501] Note: ED 50 is the median effective dose.

[0502] 7.2. MK-801-Induced Hyperactivity Test in Mice

[0503] 7.2.1 Test Method

[0504] After stratifying the mice by body weight, they were randomly divided into a model control group, a blank control group, and each dosing group. After administering the test article (or reference article), the mice were placed in an open-field activity box (a black polyethylene box with a specification of 29 cm × 29 cm × 30 cm) to adapt. 1 hour after gavage administration, 0.3 mg / kg of MK-801 was intraperitoneally injected, and then the mice were placed in the open-field activity box for video recording. The video recording time was 60 minutes. After the video recording ended, video analysis was performed to evaluate the activity of the mice.

[0505] 7.2.2 Test Results

[0506] The results of this test showed that the ED of pimaserine in inhibiting MK-801-induced hyperactivity behavior in mice 50 was 3.288 mg / kg, and the ED of NH-K-A19005, NH-K-A19006, and NH-K-A19012 in inhibiting MK-801-induced hyperactivity behavior in mice 50 were 1.01, 0.2648, and 3.728 mg / kg respectively, indicating that the compounds of the present invention have better effects against mental diseases and better drug efficacy. The detailed results are shown in the following table.

[0507] Table 9 ED of Compounds Such as Pimaserine in Inhibiting MK-801-Induced Hyperactivity Behavior in Mice 50

[0508] Compound <![CDATA[ED 50 (mg / kg)]]> Pimavanserin 3.288 NH-K-A19005 1.01 NH-K-A19006 0.2648 NH-K-A19012 3.728

[0509] Note: ED 50 Median effective dose.

[0510] 7.3. DOI ((±)-1-(2,5-Dimethoxy-4-iodophenyl)-2-aminopropane hydrochloride, (±)-2,5-Dimethoxy-4-iodoamphetamine hydrochloride) Induced PPI Injury Effect Test in Rats

[0511] 7.3.1 Test Method

[0512] Intervention Test:

[0513] Thirty minutes after intragastric administration (or administration of vehicle), and thirty minutes after subcutaneous injection of 0.5 mg / kg DOI into the neck of the animal, that is, sixty minutes after intragastric administration, the rats were placed in a startle reflex test chamber for testing.

[0514] The test method was referred to the literature and preliminary experiments, and the specific process was as follows: First, there was a 5-minute adaptation time (background sound of 62 dB). After the adaptation period, 5 separate startle reflex stimuli were given (block1, and the results were not included in the analysis to reduce the initial response of the animal to a plateau level). Subsequently, 4 different types of tests (block2) were presented in a pseudo-random manner, which were: 1) separate startle reflex stimulus (pulse-alone, 120 dB, lasting 20 ms); 2) separate prepulse stimulus 13 dB higher than the background sound (prepulse-alone, 75 dB, lasting 20 ms); 3) combined test of prepulse stimulus and startle reflex stimulus (prepulse + pulse, each lasting 20 ms, with an interval of 100 ms between them); 4) no-stimulus test with only background sound (no stimulus). Each test (trial) was presented 5 times, and the average interval between each test was 20 s (10 - 30 s).

[0515] The response amplitude of the separate startle reflex stimulus or the combined prepulse and startle reflex stimulus was expressed as the AVG (a special unit of the instrument) value, and the AVG value indirectly reflected the magnitude of the cowering response of the rat body.

[0516] Evaluation Index: PPI% = (1 - response amplitude of combined prepulse and startle reflex stimulus / response amplitude of separate startle reflex stimulus) × 100. The larger the value, the deeper the degree of inhibition.

[0517] Administration Test:

[0518] For the animals in each group above, the 0.5 mg / kg DOI was changed to normal saline (NS), and other operations including administration and testing were the same as those in the intervention test.

[0519] 7.3.2 Experimental Results

[0520] Prepulse inhibition (PPI) refers to the inhibitory effect of a weak stimulus that appears before a strong startle reflex stimulus (30 - 500 ms) on the startle reflex. Research has shown that the neural nuclei and pharmacological mechanisms that regulate PPI in humans and rodents are very similar, so it is a cross-species behavioral index. Clinical studies have also found that patients with schizophrenia have impaired PPI, and some antipsychotics can improve this. Based on these characteristics, this model has been widely used in studying the pathogenesis of schizophrenia and the pharmacological effects of antipsychotics, and also as a tool for screening antipsychotics, especially for predicting the efficacy against the negative symptoms and cognitive deficits of schizophrenia.

[0521] In this experiment, DOI at a dose of 0.5 mg / kg in rats significantly disrupted PPI (P < 0.05), and NH-K-A19006 at 0.3 mg / kg, NH-K-A19005 at 3 mg / kg, and pimozide at 3 mg / kg significantly reversed the PPI impairment induced by DOI in rats (P < 0.05), as shown in the following table. In the table, "MEAN" represents the mean value, "SD" represents the standard deviation, and "P" represents the P value. The values in the table represent the reversal of impairment, and the larger the value, the more the symptoms are alleviated compared with the model group.

[0522] This study showed that NH-K-A19006 at 0.3 mg / kg and NH-K-A19005 at 3 mg / kg significantly reversed the PPI impairment induced by DOI in rats, and had no significant effect on normal rats at these doses, suggesting that NH-K-A19005 and NH-K-A19006 are effective against mental diseases, and are effective against the negative symptoms and cognitive deficits of schizophrenia.

[0523] Table 10 Effects of NH-K-A19006 on PPI impairment induced by 0.5 mg / kg DOI in rats

[0524]

[0525]

[0526] Note: Model group vs blank group; each drug administration group vs model group.

[0527] Table 11 Effects of pimozide and NH-K-A19005 on PPI impairment induced by 0.5 mg / kg DOI in rats

[0528]

[0529] Note: Model group vs blank group; each drug administration group vs model group.

[0530] Table 12 Effects of NH-K-A19006 on PPI in normal rats

[0531]

[0532]

[0533] Note: Each drug administration group vs. the blank group.

[0534] Table 13 Effects of pimaserine and NH-K-A19005 on prepulse inhibition (PPI) in normal rats

[0535]

[0536] Note: Each drug administration group vs. the blank group.

[0537] Test Example 8 Experiment on the effect of DOI-induced head-tossing behavior in mice

[0538] Experimental method:

[0539] After stratifying ICR mice by body weight, they were randomly divided into: blank group, model group, A19006 citrate and phosphate: 0.003, 0.01, 0.03, 0.1 mg / kg groups, pimaserine L-tartrate: 0.1, 0.3, 1, 3 mg / kg groups, with 16 animals in each group (8 males and 8 females). 1 hour after intragastric administration of the test article (or reference article), the animals were placed in a beaker (13 cm in diameter and 19 cm in height) lined with fresh bedding, and the modeling drug DOI (10 mL / kg body weight) was intraperitoneally injected at a dose of 1 mg / kg. The number of head-tossing movements of the mice within 0 - 20 minutes after intraperitoneal injection of DOI was recorded. Head-tossing behavior was defined as rapid rotational twitching or wet dog-like shaking of the mouse's head, and this movement should be distinguished from normal grooming or exploratory behaviors.

[0540] Experimental results: The specific experimental results are shown in Table 14. Under the conditions of this experiment, the ED 50 values of A19006 citrate and phosphate in inhibiting head-tossing behavior in mice were 0.018 and 0.012 mg / kg respectively, and the ED 50 value of pimaserine L-tartrate in inhibiting head-tossing behavior in mice was 0.73 mg / kg, indicating that both A19006 citrate and phosphate had good activity and were superior to pimaserine L-tartrate.

[0541] Table 14 ED 50

[0542] #imgpt137#

[0543] Those skilled in the art will appreciate that many modifications and variations of the present invention can be made without departing from its spirit and scope. The specific embodiments described herein are provided by way of example only and are not meant to limit in any way. The true scope and spirit of the present invention are shown by the appended claims, and the description and examples are merely illustrative.

Claims

1. A salt of a compound represented by the following general formula I Wherein: n1 and n2 are integers from 1 to 3; R1 is selected from straight-chain or branched C1-C8 alkyl, C2-C8 alkenyl and C2-C8 alkynyl, and the alkyl, alkenyl and alkynyl are each independently and optionally substituted with substituents selected from halogen and C1-C8 haloalkyl; R2 is selected from hydrogen, halogen and C1-C8 haloalkyl; R3, R4, R5 and R6 are each independently selected from hydrogen, halogen and C1-C8 haloalkyl; R7 is selected from linear or branched C1-C8 alkyl, C3-C 10 cycloalkyl, and R8 and R9 are each independently selected from linear or branched C1-C8 alkyl, and the alkyl and cycloalkyl are optionally substituted with substituents selected from halogen and C1-C8 haloalkyl; Z is selected from CH2, O and NH; W is selected from CH and N; Q is N.

2. The salt of the compound represented by General Formula I as described in Claim 1, characterized in that: The salt is a phosphate or citrate, as shown in Formula II and Formula III: Wherein: y is 0.5, 1, 1.5, 2, 2.5 or 3; n1 and n2 are integers from 1 to 3; R1 is selected from straight-chain or branched C1-C8 alkyl, C2-C8 alkenyl and C2-C8 alkynyl, and the alkyl, alkenyl and alkynyl are each independently and optionally substituted with substituents selected from halogen and C1-C8 haloalkyl; R2 is selected from hydrogen, halogen and C1-C8 haloalkyl; R3, R4, R5 and R6 are each independently selected from hydrogen, halogen and C1-C8 haloalkyl; R7 is selected from linear or branched C1-C8 alkyl, C3-C 10 cycloalkyl and R8 and R9 are each independently selected from linear or branched C1-C8 alkyl, and the alkyl and cycloalkyl are optionally substituted with substituents selected from halogen and C1-C8 haloalkyl; Z is selected from CH2, O and NH; W is selected from CH and N; Q is N.

3. The salt of the compound represented by General Formula I according to claim 2, characterized in that: y is 0.5, 1, 1.5 or 2.

4. The salt of the compound represented by the general formula I according to any one of claims 1-3, characterized in that: The straight-chain or branched C1-C8 alkyl is a straight-chain or branched C1-C5 alkyl; and / or The C2-C8 alkenyl is a C2-C5 alkenyl; and / or The C2-C8 alkynyl is a C2-C5 alkynyl; and / or The C1-C8 haloalkyl is a C1-C5 haloalkyl; and / or The C3-C 10 cycloalkyl group is a C3-C6 cycloalkyl group.

5. The salt of the compound represented by General Formula I as described in Claim 4, characterized in that: The straight-chain or branched C1-C8 alkyl is a straight-chain or branched C1-C3 alkyl.

6. The salt of the compound represented by the general formula I according to claim 4, characterized in that: The halogen is selected from fluorine, chlorine, bromine and iodine; and / or The straight-chain or branched C1-C5 alkyl is selected from methyl, ethyl, propyl, isopropyl, butyl, isobutyl, pentyl and isopentyl; and / or The C3-C6 cycloalkyl is selected from cyclopropyl, cyclobutyl and cyclopentyl.

7. The salt of the compound represented by the general formula I according to claim 6, characterized in that: The halogen is fluorine or chlorine; and / or The straight-chain or branched C1-C5 alkyl is selected from methyl, ethyl, propyl, isopropyl and isobutyl; and / or The C3-C6 cycloalkyl is selected from cyclopropyl and cyclobutyl; The said is 8. The salt of the compound represented by General Formula I as described in Claim 1, characterized in that, Including phosphates or citrates selected from the following compounds:

9. A citrate or phosphate of A19006, characterized in that, The structure is as shown in Formula IV and V, 10. A polymorphic form I of A19006 citrate as shown in formula IV of claim 9, characterized in that, Using Cu-Kα radiation, an X-ray powder diffraction pattern expressed in terms of diffraction angle 2θ±0.2° is obtained, which shows characteristic peaks at 10.03, 12.62, 13.69, 14.68, 16.44, 17.19, 19.10, 20.27, 21.77, wherein the error range of 2θ of each characteristic peak is ±0.

2.

11. The crystalline form I according to claim 10, characterized in that, The X-ray powder diffraction pattern of Form I shows characteristic peaks at 17.67, 19.57, 21.30, 23.78, 24.03, 25.57, 26.22 expressed as angles of 2θ±0.2°, wherein the error range of 2θ for each characteristic peak is ±0.

2.

12. The crystalline form I according to claim 11, wherein The X-ray powder diffraction pattern of Form I is substantially as shown in Figure 1.

13. The crystalline form I according to any one of claims 10-12, characterized in that, The melting endothermic peak of the DSC of Form I is selected from 164.7–170.0 °C.

14. The crystalline form I according to claim 13, wherein The melting endothermic peak of the DSC of Form I is 167.7 °C.

15. A pharmaceutical composition, characterized in that Form I of the salt according to any one of claims 1-9 or the A19006 citrate according to any one of claims 10-14 and a pharmaceutically acceptable carrier.

16. Use of the salt according to any one of claims 1-9, Form I of the A19006 citrate according to any one of claims 10-14 or the pharmaceutical composition according to claim 15 in the preparation of a medicament for treating mental disorders.

17. The application according to claim 16, characterized in that, The mental disorders are schizophrenia, schizoaffective psychosis, Parkinson's disease, behavioral disturbances and psychosis associated with dementia (BPSD), delusional disorder, acute transient psychotic disorder, depressive disorder, bipolar disorder, generalized anxiety disorder, panic disorder, obsessive-compulsive disorder, social phobia disorder, agoraphobia disorder, post-traumatic stress disorder.

18. The application according to claim 17, wherein The mental disorders are Parkinson's disease, behavioral disturbances and psychosis associated with dementia (BPSD) or schizophrenia.

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