Steroid compound as well as application and preparation method thereof

Abstract

The invention relates to a steroidal compound as well as application and a preparation method thereof. The compound is expected to be capable of effectively treating psychiatric and neurological diseases, and has good active efficacy, pharmacokinetic (PK) performance, oral bioavailability, stability, safety, clearance rate and / or metabolic performance and the like.

Description

[0001] This application is a divisional application of the patent filed on January 7, 2020, with application number 202010015190.X and invention title 'steroidal compounds, uses and preparation methods thereof'. Technical Field

[0002] This invention relates to the field of pharmaceuticals, specifically to steroidal compounds, their uses, and methods of preparation. Technical Background

[0003] Neuropsychiatric disorders, including anxiety disorders, depression, schizophrenia, etc., rank first in the total disease burden in my country, accounting for about 20% of the total disease burden (Wang Juncheng et al., China Health Service Management, 2009(5):348-350). With the continuous development of social modernization, the pace of work and the increase in life pressure have led to a significant increase in patients with various neuropsychiatric disorders, and the progression of symptoms has accelerated significantly. Therefore, the development, research and production of drugs for neuropsychiatric disorders have become more urgent. At the same time, clinical data show that many patients are troubled by multiple neuropsychiatric disorders at the same time. The high comorbidity rate of neuropsychiatric disorders makes clinical treatment face greater challenges.

[0004] In animals, GABA (gamma-aminobutyric acid) exists only in nerve tissue, and immunological studies have shown that its highest concentration is found in the substantia nigra of the brain. GABA is an important inhibitory neurotransmitter that has been extensively studied; it participates in various metabolic activities and possesses high physiological activity. The GABA system is a major inhibitory signaling pathway in the brain and central nervous system, playing a crucial role in regulating central nervous system function. The GABA(A) receptor (GABAAR) is an ionotropic receptor and ligand-gated ion channel. Its endogenous ligand, γ-aminobutyric acid (GABA), is the main inhibitory neurotransmitter in the central nervous system. After activation, the GABA(A) receptor selectively allows Cl... - Through its pores. Cl -Action potentials flow out of neurons when the internal voltage is below the resting potential and flow into neurons when the internal voltage is above the resting potential (i.e., -75mV). This successfully reduces the chance of action potentials and inhibits neurotransmission. Anxiety and depression have a very high comorbidity rate and are considered to have overlapping and co-occurring phenomena. Anxiety and depression are also important mood symptoms of schizophrenia. Research has confirmed that the GABAergic system and GABA(A) receptors play important roles in the pathogenesis of these three diseases, indicating that a pathophysiological process related to the GABAergic system may be one of the common determinants of these three diseases. The GABAergic system and GABA(A) receptors have been shown to participate in the pathological processes of anxiety, depression, and schizophrenia at the molecular, preclinical, and clinical levels, and GABA(A) receptors have long been considered an important drug target for the treatment of these diseases.

[0005] Clinical trials have confirmed that, in addition to binding to GABA itself to form receptor complexes (GRCs) and altering brain excitability, GABA(A) receptors can also bind to other small molecule compounds with specific structures, such as barbiturates (brand name: Socotoxin) and benzodiazepines. Drugs such as diazepam (trade name) bind to specific allosteric sites on the GABA(A) receptor to produce their therapeutic effects. In addition, studies have shown that there are also unique sites on the GABA(A) receptor for steroidal compounds (Lan, NC et al., Neurochem. Res. 16:347-356 (1991)).

[0006] Neuroactive steroids can be produced endogenously. The most potent endogenous neuroactive steroids are 3α-hydroxy-5-reductorpregnane-20-one and 3α-21-dihydroxy-5-reductorpregnane-20-one, which are metabolites of the hormonal steroids progesterone and deoxycorticosterone, respectively. The ability of these steroid metabolites to alter brain excitability was recognized in 1986 (Majewska, M.D. et al., Science 232:1004-1007 (1986); Harrison, N. Letal., J. Pharmacol. Exp. Ther. 241:346-353 (1987)). The ovarian hormone progesterone and its metabolites have been shown to have a profound effect on brain excitability (Backstrom, T. et al., Acta Obstet. Gynecol. Scand. Suppl. 130:19-24 (1985); Pfaff, D. Wand McEwen, B. Setal. Science 219:808-814 (1983); Gyermek et al., J Med Chem. 11:117 (1968); Lambert, J. et al., Trends Pharmacol. Sci. 8:224-227 (1987)). The levels of progesterone and its metabolites change with the phases of the menstrual cycle. Numerous studies have demonstrated that progesterone and its metabolites levels decrease before the onset of menstruation. The recurring monthly physical symptoms that precede the onset of menstruation have also been documented in numerous studies. These symptoms, which have become associated with premenstrual syndrome (PMS), include stress (tension), anxiety, and migraines (Dalton, K., Premenstrual Syndrome and Progesterone Therapy, 2nd edition, Chicago Year book, Chicago (1984)). Subjects with PMS experience symptoms that recur monthly, present before menstruation and absent after menstruation.

[0007] Other studies have found that decreased progesterone levels are associated with an increased frequency of seizures in women with epilepsy, specifically menstrual epilepsy (Laidlaw, J., Lancet, 1235-1237 (1956)). A more direct association has been observed with decreased progesterone metabolites (Rosciszewska et al., J. Neurol. Neurosurg. Psych. 49:47-51 (1986)). Furthermore, the incidence of primary generalized absence seizures has been correlated with the incidence of premenstrual symptoms in patients with this condition (Backstrom, T. et al., J. Psychosom. Obstet. Gynaecol. 2:8-20 (1983)). The steroid deoxycorticosterone has been found to be effective in treating patients with epilepsy associated with the menstrual cycle (Aird, R.B. and Gordan, G., J. Amer. Med. Soc. 145:715-719 (1951)).

[0008] In addition to the conditions mentioned above, postpartum depression (PND) is also associated with low progesterone levels. Following delivery, progesterone levels drop sharply and immediately, leading to the onset of PND. Symptoms of PND range from mild depression to psychosis requiring hospitalization. PND is also associated with severe anxiety and irritability. Depression associated with PND cannot be treated with classic antidepressants, and women who experience PND have an increased probability of developing PMS (Dalton, K., Premenstrual Syndrome and Progesterone Therapy, 2nd edition, Chicago Yearbook, Chicago (1984)). These observations more or less suggest the key role of progesterone and deoxycorticosterone, especially their metabolites, in regulating brain excitability, with related clinical indications including menstrual epilepsy, PMS, and PND.

[0009] Many studies are based on GABA(A) receptors in order to obtain drugs that can effectively treat related diseases. CN103958540A, CN10533928A, and others have disclosed a series of neurosteroid compounds for the treatment of neuropsychiatric diseases. Summary of the Invention

[0010] One objective of this invention is to provide a steroid compound with improved efficacy, enabling more effective treatment of neuropsychiatric disorders. The compounds of this invention are expected to offer favorable activity, pharmacokinetic (PK) properties, oral bioavailability, formulatability, stability, safety, clearance, and / or metabolic performance.

[0011] To achieve the above objectives, the present invention provides a compound of formula I and a pharmaceutically acceptable salt thereof:

[0012]

[0013] R1 is selected from substituted or unsubstituted five-membered aromatic heterocycles, substituted or unsubstituted thiazolyl groups, and substituted or substituted benzothiophene groups;

[0014] R2 is a hydrogen atom or a methyl group.

[0015] The substituted or unsubstituted five-membered aromatic heterocycle is preferably selected from...

[0016] or double substitution

[0017] or tri-substitution

[0018] Where X is O or S;

[0019] R3, R4, and R5 are each independently hydrogen, substituted or unsubstituted C1-6 alkyl, halogen, -NO2, -CN, -OR, -N(R)2, -C(=O)R, -C(=O)OR, -OC(=O)R, -OC(=O)OR, -C(=O)N(R)2, -N(R)C(=O)R, -OC(=O)N(R)2, -N(R)C(=O)OR, -N(R)C(=O)N(R)2, -SR, -S(O)R, for example -S(=O)R, -S(=O)2R, -S(=O)2OR, -OS(=O)2R, -S(=O)2N(R)2, -N(R)S(=O)2R, substituted or unsubstituted C2-6 alkenyl, substituted or unsubstituted C2-6 alkynyl.

[0020] The substituted or unsubstituted thiazolyl group is preferably from a monosubstituted group.

[0021] or double substitution

[0022] R3 and R4 are each independently hydrogen, substituted or unsubstituted C1-6 alkyl, halogen, -NO2, -CN, -OR, -N(R)2, -C(=O)R, -C(=O)OR, -OC(=O)R, -OC(=O)OR, -C(=O)N(R)2, -N(R)C(=O)R, -OC(=O)N(R)2, -N(R)C(=O)OR, -N(R)C(=O)N(R)2, -SR, -S(O)R, for example -S(=O)R, -S(=O)2R, -S(=O)2OR, -OS(=O)2R, -S(=O)2N(R)2, -N(R)S(=O)2R, substituted or unsubstituted C2-6 alkenyl, substituted or unsubstituted C2-6 alkynyl.

[0023] The benzene-substituted or unsubstituted thiazolyl group is preferably selected from...

[0024] Wherein R3 is hydrogen, substituted or unsubstituted C1-6 alkyl, halogen, -NO2, -CN, -OR, -N(R)2, -C(=O)R, -C(=O)OR, -OC(=O)R, -OC(=O)OR, -C(=O)N(R)2, -N(R)C(=O)R, -OC(=O)N(R)2, -N(R)C(=O)OR, -N(R)C(=O)N(R)2, -SR, -S(O)R, for example -S(=O)R, -S(=O)2R, -S(=O)2OR, -OS(=O)2R, -S(=O)2N(R)2, -N(R)S(=O)2R, substituted or unsubstituted C2-6 alkenyl, substituted or unsubstituted C2-6 alkynyl.

[0025] The compound of formula I and its pharmaceutically acceptable salt are selected from...

[0026]

[0027]

[0028]

[0029] The present invention also provides a pharmaceutical composition comprising any of the compounds described above or their pharmaceutically acceptable salts, and pharmaceutically usable excipients.

[0030] The compounds described herein can act as GABA modulators, for example, affecting GABA(A) receptors in a positive or negative manner. As regulators of central nervous system (CNS) excitability, such compounds are expected to possess CNS activity when mediated by their ability to modulate GABA(A) receptors.

[0031] The present invention further provides the use of the above-mentioned compounds or pharmaceutical compositions in the preparation of medicaments for the prevention or treatment of nervous system diseases; wherein the nervous system diseases are preferably selected from: sleep disorders, mood disorders, schizophrenia spectrum disorders, spastic disorders, memory disorders and / or cognitive disorders, motor disorders, personality disorders, autism spectrum disorders, pain, traumatic brain injury, vascular diseases, substance abuse disorders and / or truncation syndromes or tinnitus; wherein the mood disorder is depression; preferably major depressive disorder or postpartum depression.

[0032] Furthermore, the aforementioned compounds or pharmaceutical compositions can be administered via oral, subcutaneous, intravenous, or intramuscular routes.

[0033] definition

[0034] Chemical definition

[0035] The compounds described herein may include one or more asymmetric centers and therefore may exist in a variety of isomeric forms, such as enantiomers and / or diastereomers. For example, the compounds described herein may be individual enantiomers, diastereomers, or geometric isomers, or may be in the form of mixtures of stereoisomers, including racemic mixtures and mixtures rich in one or more stereoisomers. Isomers can be separated from mixtures by methods known to those skilled in the art, including chiral high-performance liquid chromatography (HPLC) and the formation and crystallization of chiral salts; or preferred isomers may be prepared by asymmetric synthesis.

[0036] The following terms are intended to have the meanings provided below and are useful in understanding the specification and intended scope of the invention. When describing the invention, it may include compounds, pharmaceutical compositions containing said compounds, and methods for testing said compounds and compositions. Definitions of terms used in this invention can be referenced to the following description, and any portion defined below may be substituted with a number of substituents, and the corresponding definitions within their ranges listed below include such substituted portions. Unless otherwise stated, the term "substitution" is defined as follows.

[0037] "alkyl" refers to a straight-chain or branched saturated hydrocarbon group having 1 to 20 carbon atoms ("C 1-20 alkyl"). In some embodiments, the alkyl group has 1 to 12 carbon atoms ("C 1-12 alkyl"). In some embodiments, the alkyl group has 1 to 10 carbon atoms ("C 1-10 alkyl"). In some embodiments, the alkyl group has 1 to 9 carbon atoms ("C 1-9 alkyl"). In some embodiments, the alkyl group has 1 to 8 carbon atoms ("C 1-8 alkyl"). In some embodiments, the alkyl group has 1 to 7 carbon atoms ("C 1-7 alkyl"). In some embodiments, the alkyl group has 1 to 6 carbon atoms ("C 1-6 alkyl", which also refers to "lower alkyl"). In some embodiments, the alkyl group has 1 to 5 carbon atoms ("C 1-5 alkyl"). In some embodiments, the alkyl group has 1 to 4 carbon atoms ("C 1-4 alkyl"). In some embodiments, the alkyl group has 1 to 3 carbon atoms ("C 1-3 alkyl"). In some embodiments, the alkyl group has 1 to 2 carbon atoms (“C1-2 alkyl”). In some embodiments, the alkyl group has 1 carbon atom (“C1 alkyl”). In some embodiments, the alkyl group has 2 to 6 carbon atoms (“C2-6 alkyl”). Examples of C1-6 alkyl groups include: methyl (C1), ethyl (C2), n-propyl (C3), isopropyl (C3), n-butyl (C4), tert-butyl (C4), sec-butyl (C4), isobutyl (C4), n-pentyl (C5), 3-pentyl (C5), pentyl (C5), neopentyl (C5), 3-methyl-2-butyl (C5), tert-pentyl (C5), and n-hexyl (C6). Other examples of alkyl groups include n-heptyl (C7), n-octyl (C8), and so on. Unless otherwise stated, each alkyl group is optionally substituted independently, i.e., unsubstituted (“unsubstituted alkyl”) or substituted with one or more substituents (“substituted alkyl”); for example, 1 to 5 substituents, 1 to 3 substituents, or 1 substituent. In some embodiments, the alkyl group is an unsubstituted C1-10 alkyl group (e.g., -CH3). In some embodiments, the alkyl group is a substituted C1-10 alkyl group.

[0038] "Alkenyl" refers to a straight-chain or branched hydrocarbon group ("C 2-20 alkenyl") having 2 to 20 carbon atoms, one or more carbon-carbon double bonds (e.g., 1, 2, 3, or 4 carbon-carbon double bonds), and optionally one or more carbon-carbon triple bonds (e.g., 1, 2, 3, or 4 carbon-carbon triple bonds). In some embodiments, the alkenyl group does not contain any triple bonds. In some embodiments, the alkenyl group has 2 to 10 carbon atoms ("C 2-10 alkenyl"). In some embodiments, the alkenyl group has 2 to 9 carbon atoms ("C 2-9 alkenyl"). In some embodiments, the alkenyl group has 2 to 8 carbon atoms ("C 2-8 alkenyl"). In some embodiments, the alkenyl group has 2 to 7 carbon atoms ("C 2-7 alkenyl"). In some embodiments, the alkenyl group has 2 to 6 carbon atoms ("C 2-6 alkenyl"). In some embodiments, the alkenyl group has 2 to 5 carbon atoms ("C 2-5 alkenyl"). In some embodiments, the alkenyl group has 2 to 4 carbon atoms (“C 2-4 alkenyl”). In some embodiments, the alkenyl group has 2 to 3 carbon atoms (“C 2-3 alkenyl”). In some embodiments, the alkenyl group has 2 carbon atoms (“C 2 alkenyl”). One or more carbon-carbon double bonds may be internal (e.g., in 2-butenyl) or terminal (e.g., in 1-butenyl). Examples of C 2-4 alkenyl groups include: vinyl (C 2), 1-propenyl (C 3), 2-propenyl (C 3), 1-butenyl (C 4), 2-butenyl (C 4), butadienyl (C 4), etc. Examples of C 2-6 alkenyl groups include: the C 2-4 alkenyl groups described above, as well as pentenyl (C 5), pentadienyl (C 5), hexenyl (C 6), etc. Other examples of alkenyl groups include heptenyl (C 7), octenyl (C 8), octetrinyl (C 8), and so on. Unless otherwise stated, each alkenyl group is optionally substituted independently, i.e., unsubstituted (“unsubstituted alkenyl”) or substituted with one or more substituents (“substituted alkenyl”); for example, 1 to 5 substituents, 1 to 3 substituents, or 1 substituent. In some embodiments, the alkenyl group is an unsubstituted C 2-10 alkenyl group. In some embodiments, the alkenyl group is a substituted C 2-10 alkenyl group.

[0039] "Alynyl" refers to a straight-chain or branched hydrocarbon group ("C 2-20 ynyl") having 2 to 20 carbon atoms, one or more carbon-carbon triple bonds (e.g., 1, 2, 3, or 4 carbon-carbon triple bonds), and optionally one or more carbon-carbon double bonds (e.g., 1, 2, 3, or 4 carbon-carbon double bonds). In some embodiments, the ynyl group does not contain any double bonds. In some embodiments, the ynyl group has 2 to 10 carbon atoms ("C 2-10 ynyl"). In some embodiments, the ynyl group has 2 to 9 carbon atoms ("C 2-9 ynyl"). In some embodiments, the ynyl group has 2 to 8 carbon atoms ("C 2-8 ynyl"). In some embodiments, the ynyl group has 2 to 7 carbon atoms ("C 2-7 ynyl"). In some embodiments, the ynyl group has 2 to 6 carbon atoms ("C 2-6 ynyl"). In some embodiments, the ynyl group has 2 to 5 carbon atoms ("C 2-5 ynyl"). In some embodiments, the alkynyl group has 2 to 4 carbon atoms (“C 2-4 alkynyl”). In some embodiments, the alkynyl group has 2 to 3 carbon atoms (“C 2-3 alkynyl”). In some embodiments, the alkynyl group has 2 carbon atoms (“C 2 alkynyl”). One or more carbon triple bonds may be internal (e.g., in 2-butynyl) or terminal (e.g., in 1-butynyl). Examples of C 2-4 alkynyl groups include, but are not limited to: ethynyl (C 2), 1-propynyl (C 3), 2-propynyl (C 3), 1-butynyl (C 4), 2-butynyl (C 4), etc. Examples of C 2-6 alkenyl groups include: the aforementioned C 2-4 alkynyl groups, as well as pentyynyl (C 5), hexynyl (C 6), etc. Other examples of alkynyl groups include heptyynyl (C 7), octyynyl (C 8), etc. Unless otherwise stated, each of the ynyl groups is optionally substituted independently, i.e., unsubstituted (“unsubstituted ynyl”) or substituted with one or more substituents (“substituted ynyl”); for example, 1 to 5 substituents, 1 to 3 substituents, or 1 substituent. In some embodiments, the ynyl group is an unsubstituted C2-10 ynyl group. In some embodiments, the ynyl group is a substituted C2-10 ynyl group.

[0040] "Aryl" refers to a group ("C 6-14 aryl") having a monocyclic or polycyclic (e.g., bicyclic or tricyclic) 4n+2 aromatic ring system (e.g., having 6, 10, or 14 shared π electrons arranged in a ring) providing 6-14 ring carbon atoms and zero heteroatoms in an aromatic ring system. In some embodiments, the aryl has six ring carbon atoms ("C 6 aryl"; e.g., phenyl). In some embodiments, the aryl has ten ring carbon atoms ("C 10 aryl"; e.g., naphthyl, e.g., 1-naphthyl and 2-naphthyl). In some embodiments, the aryl has fourteen ring carbon atoms ("C 14 aryl"; e.g., anthracene). "Aryl" also includes ring systems in which the aforementioned aryl ring is fused with one or more carbocyclic or heterocyclic groups, wherein the atomic group or connecting point is on the aryl ring, in which case the number of carbon atoms continues to represent the number of carbon atoms in the aryl ring system. Typical aryl groups include, but are not limited to, those derived from the following groups: anthracene, acenaphthene, phenanthrylene, anthracene, azulene, benzene, fluoranthene, phenanthracene, indene, indene, naphthalene, octaphenyl, octane, octane, oleophane, pentadiene, pentane, pentaphenyl, pentaphenyl, pentaphenyl, perylene, phenanthracene, phenanthrene, phenanthrene, heptaphenyl, pyrene, pinane, rubigin, benzophenanthrene, and ternaphthalene. Specifically, aryl groups include phenyl, naphthyl, indene, and tetrahydronaphthyl. Unless otherwise stated, each aryl group is optionally substituted independently, i.e., unsubstituted (“unsubstituted aryl”) or substituted by one or more substituents (“substituted aryl”). In some embodiments, the aryl group is an unsubstituted C6-14 aryl group. In some embodiments, the aryl group is a substituted C6-14 aryl group. In some embodiments, the aryl group is substituted with one or more groups selected from halogen, C1-C8 alkyl, C1-C8 haloalkyl, cyano, hydroxyl, C1-C8 alkoxy, and amino.

[0041] "Heteroaryl" refers to a 4n+2 aromatic ring system (e.g., having 6 or 10 shared π electrons arranged in a ring) having a 5-10 membered monocyclic or bicyclic ring providing a cyclic carbon atom and 1-4 cyclic heteroatoms in the aromatic ring system, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur ("5-10 membered heteroaryl"). In heteroaryls containing one or more nitrogen atoms, the linkage can be a carbon or nitrogen atom, provided the valence allows. A heteroaryl bicyclic system may include one or more heteroatoms in one or both rings. "Heteroaryl" includes a ring system in which the aforementioned heteroaryl ring is fused with one or more carbocyclic or heterocyclic groups, wherein the linkage is on the heteroaryl ring, and in such cases, the number of ring members continues to represent the number of ring members in the heteroaryl ring system. "Heteroaryl" also includes ring systems in which the aforementioned heteroaryl ring is fused with one or more aryl groups, wherein the connection point is on the aryl or heteroaryl ring, and in such cases, the number of ring members represents the number of ring members in the fused (aryl / heteroaryl) ring system. A bicyclic heteroaryl (e.g., indolyl, quinolinyl, carbazolyl, etc.) in which one ring does not contain a heteroatom can have the connection point on either ring, i.e., on the ring with the heteroatom (e.g., 2-indolyl) or the ring without the heteroatom (e.g., 5-indolyl). In some embodiments, the heteroaryl is a 5-10 membered aromatic ring system having a ring carbon atom and 1-4 ring heteroatoms (present in the aromatic ring system), wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur ("5-10 membered heteroaryl"). In some embodiments, the heteroaryl group is a 5-8 membered aromatic ring system having a cyclic carbon atom and 1-4 cyclic heteroatoms (present in an aromatic ring system), wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur (“5-8 membered heteroaryl”). In some embodiments, the heteroaryl group is a 5-6 membered aromatic ring system having a cyclic carbon atom and 1-4 cyclic heteroatoms (present in an aromatic ring system), wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur (“5-6 membered heteroaryl”). In some embodiments, the 5-6 membered heteroaryl group has 1-3 cyclic heteroatoms selected from nitrogen, oxygen, and sulfur. In some embodiments, the 5-6 membered heteroaryl group has 1-2 cyclic heteroatoms selected from nitrogen, oxygen, and sulfur. In some embodiments, the 5-6 membered heteroaryl group has 1 cyclic heteroatom selected from nitrogen, oxygen, and sulfur. Unless otherwise stated, each heteroaryl group is optionally substituted independently, i.e., unsubstituted (“unsubstituted heteroaryl”) or substituted with one or more substituents (“substituted heteroaryl”). In some embodiments, the heteroaryl group is an unsubstituted 5-14 membered heteroaryl group. In some embodiments, the heteroaryl group is a substituted 5-14 membered heteroaryl group. 5-membered heteroaryl groups containing one heteroatom include, but are not limited to: pyrrole, furanyl, and thiophene. 5-membered heteroaryl groups containing two heteroatoms include, but are not limited to: imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, and isothiazolyl. 5-membered heteroaryl groups containing three heteroatoms include, but are not limited to: triazolyl, oxadiazolyl, and thiazolyl.Five-membered heteroaryl groups containing four heteroatoms include, but are not limited to, tetrazolyl. Six-membered heteroaryl groups containing one heteroatom include, but are not limited to, pyridyl. Six-membered heteroaryl groups containing two heteroatoms include, but are not limited to, pyridazinyl, pyrimidinyl, and pyrazinyl. Six-membered heteroaryl groups containing three or four heteroatoms include, but are not limited to, triazinyl and tetraazinyl. Seven-membered heteroaryl groups containing one heteroatom include, but are not limited to, azirmonoheptatrienyl, oxadiazinyl, and thioheptatrienyl. 5,6-Bicyclic heteroaryl groups include, but are not limited to, indolyl, isoyindolyl, indazole, benzotriazolyl, benzothiopheneyl, isobenzothiopheneyl, benzofuranyl, benzoisofuranyl, benzoimidazolyl, benzoxazolyl, benzoisoxazolyl, benzoxadiazolyl, benzothiazolyl, benzoisothiazolyl, benzothiadiazolyl, indazinyl, and purinel. 6,6-Bicyclic heteroaryl groups include, but are not limited to: naphthidyl, pteridyl, quinolinyl, isoquinolinyl, zolinyl, quinoxalinyl, phthalazinyl, and quinazolinyl.

[0042] "Carbocyclic group" or "carbocyclic" refers to a non-aromatic cyclic hydrocarbon group ("C 3-10 carbocyclic group") having 3 to 10 ring carbon atoms and zero heteroatoms in a non-aromatic ring system. In some embodiments, the carbocyclic group has 3 to 8 ring carbon atoms ("C 3-8 carbocyclic group"). In some embodiments, the carbocyclic group has 3 to 6 ring carbon atoms ("C 3-6 carbocyclic group"). In some embodiments, the carbocyclic group has 3 to 6 ring carbon atoms ("C 3-6 carbocyclic group"). In some embodiments, the carbocyclic group has 5 to 10 ring carbon atoms ("C 5-10 carbocyclic group"). Exemplary C3-6 carbon cyclogroups include, but are not limited to: cyclopropyl (C3), cyclopropenyl (C3), cyclobutyl (C4), cyclobutenyl (C4), cyclopentyl (C5), cyclopentenyl (C5), cyclohexyl (C6), cyclohexenyl (C6), cyclohexadienyl (C6), etc. Exemplary C3-8 carbon cyclogroups include, but are not limited to: the above-mentioned C3-6 carbon cyclogroups, as well as cycloheptyl (C7), cycloheptenyl (C7), cycloheptadienyl (C7), cyclohepttrienyl (C7), cyclooctyl (C8), cyclooctenyl (C8), bicyclo[2.2.1]heptyl (C7), bicyclo[2.2.2]octyl (C8), etc. Exemplary C3-10 carbocyclic groups include, but are not limited to, the aforementioned C3-8 carbocyclic groups, as well as cyclononyl (C9), cyclononenyl (C9), cyclodecyl (C10), cyclodecenyl (C10), octahydro-1H-indenyl (C9), decahydronaphthyl (C10), spiro[4.5]decyl (C10), and so on. As illustrated in the foregoing examples, in some embodiments, the carbocyclic group is a monocyclic (“monocyclic carbocyclic”) or a carbocyclic group comprising a fused ring system, a bridged ring system, or a spirocyclic system, such as a bicyclic system (“bicyclic carbocyclic”), and may be saturated or may be partially unsaturated. “Carbocyclic” also includes a ring system in which the aforementioned carbocyclic ring is fused with one or more aryl or heteroaryl groups, wherein the connection point is on the carbocyclic ring, and in such cases, the number of carbons continues to represent the number of carbons in the carbocyclic system. Unless otherwise stated, each carbocyclic group is independently optionally substituted, i.e., unsubstituted (“unsubstituted carbocyclic”) or substituted with one or more substituents (“substituted carbocyclic”). In some embodiments, the carbocyclic group is an unsubstituted C3-10 carbocyclic group. In some embodiments, the carbocyclic group is a substituted C3-10 carbocyclic group. In some embodiments, “carbocyclic” is a monocyclic saturated carbocyclic group having 3 to 10 ring carbon atoms (“C3-10 cycloalkyl”). In some embodiments, the cycloalkyl group has 3 to 8 ring carbon atoms (“C3-8 cycloalkyl”). In some embodiments, the cycloalkyl group has 3 to 6 ring carbon atoms (“C3-6 cycloalkyl”). In some embodiments, the cycloalkyl group has 5 to 6 ring carbon atoms (“C5-6 cycloalkyl”).In some embodiments, the cycloalkyl group has 5 to 10 cyclic carbon atoms (“C 5-10 cycloalkyl”). Examples of C5-6 cycloalkyl groups include cyclopentyl (C 5) and cyclohexyl (C 6). Examples of C 3-6 cycloalkyl groups include the aforementioned C 5-6 cycloalkyl groups, as well as cyclopropyl (C 3) and cyclobutyl (C 4). Examples of C 3-8 cycloalkyl groups include the aforementioned C 3-6 cycloalkyl groups, as well as cycloheptyl (C 7) and cyclooctyl (C 8). Unless otherwise stated, each cycloalkyl group is independently unsubstituted (“unsubstituted cycloalkyl”) or substituted with one or more substituents (“substituted cycloalkyl”). In some embodiments, the cycloalkyl group is an unsubstituted C 3-10 cycloalkyl group. In some embodiments, the cycloalkyl group is a substituted C 3-10 cycloalkyl group.

[0043] A "heterocyclic group" or "heterocycle" refers to a non-aromatic ring system consisting of 3 to 10 members, having a ring carbon atom and 1 to 4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, sulfur, boron, phosphorus, and silicon ("3-10 membered heterocyclic groups"). In heterocyclic groups containing one or more nitrogen atoms, the bonding point can be either a carbon or nitrogen atom, provided the valence allows. Heterocyclic groups can be monocyclic ("monocyclic heterocyclic group") or fused ring, bridged ring, or spirocyclic systems, such as bicyclic systems ("bicyclic heterocyclic group"), and can be saturated or partially unsaturated. A bicyclic heterocyclic system can include one or more heteroatoms in one or both rings. "Heterocyclic group" also includes ring systems in which the aforementioned heterocyclic ring is fused with one or more carbocyclic groups, wherein the connection point is on the carbocyclic or heterocyclic ring, or ring systems in which the aforementioned heterocyclic ring is fused with one or more aryl or heteroaryl groups, wherein the connection point is on the heterocyclic ring, and in such cases, the number of ring members continues to represent the number of ring members in the heterocyclic ring system. Unless otherwise stated, each of the heterocyclic groups is independently optionally substituted, i.e., unsubstituted ("unsubstituted heterocyclic group") or substituted by one or more substituents ("substituted heterocyclic group"). In some embodiments, the heterocyclic group is an unsubstituted 3-10 membered heterocyclic group. In some embodiments, the heterocyclic group is a substituted 3-10 membered heterocyclic group. In some embodiments, the heterocyclic group is a 5-10 membered non-aromatic ring system having a ring carbon atom and 1-4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, sulfur, boron, phosphorus, and silicon ("5-10 membered heterocyclic group"). In some embodiments, the heterocyclic group is a 5-8 membered non-aromatic ring system having a cyclic carbon atom and 1-4 cyclic heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur (“5-8 membered heterocyclic group”). In some embodiments, the heterocyclic group is a 5-6 membered non-aromatic ring system having a cyclic carbon atom and 1-4 cyclic heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur (“5-6 membered heterocyclic group”). In some embodiments, the 5-6 membered heterocyclic group has 1-3 cyclic heteroatoms selected from nitrogen, oxygen, and sulfur. In some embodiments, the 5-6 membered heterocyclic group has 1-2 cyclic heteroatoms selected from nitrogen, oxygen, and sulfur. In some embodiments, the 5-6 membered heterocyclic group has one cyclic heteroatom selected from nitrogen, oxygen, and sulfur.

[0044] When used to describe a compound or a group present on a compound, "hetero" means that one or more carbon atoms in the compound or group have been replaced by nitrogen, oxygen, or sulfur heteroatoms. "Hetero" can be applied to any of the above-mentioned hydrocarbon groups: for example, alkyl, such as heteroalkyl; cycloalkyl, such as heterocyclic; aryl, such as heteroaryl; cycloalkenyl, such as cycloheterenyl, etc.; having 1 to 5 heteroatoms, especially 1 to 3 heteroatoms.

[0045] "Alkoxy" refers to the group -OR, where R is a substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted carbocyclic, substituted or unsubstituted heterocyclic, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl. Specific alkoxy groups are methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, tert-butoxy, sec-butoxy, n-pentoxy, n-hexoxy, and 1,2-dimethylbutoxy. Specific alkoxy groups are lower alkoxy groups, i.e., having 1 to 6 carbon atoms. Further specific alkoxy groups have 1 to 4 carbon atoms. Exemplary "substituted alkoxy groups" include, but are not limited to: -O-(CH 2 -O-(CH2)t (C6-C10 aryl), -O-(CH2)t (5-10 heteroaryl), -O-(CH2)t (C3-C10 cycloalkyl), and -O-(CH2)t (4-10 heterocyclic), where t is an integer from 0 to 4, and any aryl, heteroaryl, cycloalkyl, or heterocyclic element present may be substituted by an unsubstituted C1-C4 alkyl, halogen, unsubstituted C1-C4 alkoxy, unsubstituted C1-C4 haloalkyl, unsubstituted C1-C4 hydroxyalkyl, or unsubstituted C1-C4 haloalkoxy or hydroxyl.

[0046] "Carboxyl group" refers to the group -C(O)OH.

[0047] "Cyano" refers to the group -CN.

[0048] "Halogen" or "halogen" refers to fluorine (F), chlorine (Cl), bromine (Br), and iodine (I). In some embodiments, the halogen group is fluorine or chlorine. "Hydroxy" refers to the -OH group.

[0049] "Nitro" refers to the group -NO2.

[0050] Other definitions

[0051] The term "pharmaceutically acceptable salt" refers to those salts that, within the bounds of reliable medical judgment, are suitable for contact with the tissues of humans and lower animals without excessive toxicity, irritation, allergic reactions, etc., and in proportion to a reasonable benefit / risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, Berge et al. describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences (1977) 66:1-19. Pharmaceutically acceptable salts of the compounds of this invention include salts derived from suitable inorganic and organic acids and bases. Examples of pharmaceutically acceptable non-toxic acid addition salts are salts formed by amino groups with inorganic acids, such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, and perchloric acid; or salts formed with organic acids, such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid, or malonic acid; or salts formed using methods used in the art, such as ion exchange methods. Other pharmaceutically acceptable salts include: adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentylpropionate, diglucuronate, dodecyl sulfate, ethanesulfonate, formate, fumarate, gluconate, glyceryl phosphate, glucuronate, hemisulfate, heptaate, hexanoate, hydroiodate, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, dihydroxynaphthalate, pectin ester, persulfate, 3-phenylpropionate, phosphate, picrate, pentanoate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate, etc. Pharmaceutically acceptable salts derived from suitable bases include alkali metal, alkaline earth metal, ammonium, and tetraalkylammonium salts. Representative alkali metal or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and so on. Further pharmaceutically acceptable salts, where appropriate, include non-toxic ammonium salts, quaternary ammonium salts, and amine cations formed using counterions such as halide, hydroxide, carboxyl, sulfate, phosphate, nitrate, lower alkyl sulfonate, and aryl sulfonate. Detailed Implementation

[0052] To better explain the technical solution of the present invention, the present invention provides some examples of the synthesis or biological embodiments of the compounds, which are not intended to further limit the scope of protection of the invention.

[0053] Raw materials and methods

[0054] The compounds described herein can be prepared from readily available starting materials using the following general methods and processes. It should be understood that other process conditions may be used under the given typical or preferred process conditions (i.e., reaction temperature, time, molar ratio of reactants, solvent, pressure, etc.) unless otherwise stated. Optimal reaction conditions may vary depending on the specific reactants or solvents used, but such conditions can be determined by those skilled in the art through conventional optimization.

[0055] Furthermore, it will be apparent to those skilled in the art that conventional protecting groups may be necessary to prevent certain functional groups from undergoing undesirable reactions. The selection of suitable protecting groups for specific functional groups and suitable conditions for protection and deprotection is well known in the art.

[0056] The compounds described herein can be isolated and purified using known standard methods. These methods include (but are not limited to) recrystallization, column chromatography, HPLC, or supercritical fluid chromatography (SFC). The following reaction routes are provided for the preparation details of the representative pyrazoles listed herein. The compounds described herein can be prepared by those skilled in the art of organic synthesis using known or commercially available starting materials and reagents.

[0057] The compounds of the present invention can be prepared according to the methods described in the art, using suitable reagents, raw materials and purification methods known to those skilled in the art.

[0058] Example 1: Synthesis of compound KHL

[0059]

[0060] Compound KHL-2: HBr (1.0 mL) was added to a suspension of compound KHL-1 (30.0 g, 110.3 mmol) and 10% Pd / C (1.5 g) in tetrahydrofuran (300 mL). The mixture was degassed three times with hydrogen and stirred at room temperature for 24 hours under a hydrogen atmosphere. After the reaction was complete, the mixture was filtered together with the same reaction mixture (40.0 g) and washed with tetrahydrofuran to give the crude product. The crude product was slurried with petroleum ether / acetone (1:1) to give compound KHL-2 (60.0 g, 85.1%) as a white solid.

[0061] Compound KHL-3: Iodine (2.8 g, 11.0 mmol) was added to a methanol (500 mL) solution of compound KHL-2 (30 g, 109.5 mmol), and the mixture was stirred at 60 °C for 12 hours. Then, pyridine (8.7 g, 110.0 mmol) was added. After the reaction was complete, the solvent was removed by vacuum concentration with the same reaction mixture (30.0 g), and the residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 10:1) to give a white solid compound KHL-3 (52.0 g, 74.2%).

[0062] Compound KHL-4: Ethyltriphenylphosphine bromide (192.9 g, 520.0 mmol) was added to a solution of potassium tert-butoxide (54.6 g, 487.5 mmol) in tetrahydrofuran (800 mL) at 0 °C. After stirring at 60 °C for 3 hours, a solution of compound KHL-3 (52.0 g, 162.5 mmol) in tetrahydrofuran (200 mL) was added, and stirring was continued at 60 °C for 2 hours. After the reaction was complete, the mixture was cooled to room temperature, poured into a saturated NH4Cl solution, and extracted with ethyl acetate (2 L * 3). The combined organic phases were concentrated under vacuum, and the residue was purified by silica gel column chromatography (petroleum ether) to give a white solid compound KHL-4 (48.0 g, 89.0%).

[0063] Compound KHL-5: A solution of anhydrous tetrahydrofuran (1 L) of compound KHL-4 (48.0 g, 144.6 mmol) was dissolved in 216.9 mL of BH3-THF solution (2.0 M, 433.8 mmol). After reacting at room temperature for 1 hour, the reaction mixture was cooled to 0 °C and slowly quenched successively with 216.9 mL of 10% NaOH solution and 260.3 mL of 30% H2O2 solution. The mixture was stirred at room temperature for 1 hour and extracted with ethyl acetate (2 L * 3). The combined organic layers were washed successively with 10% Na2S2O3 solution and saturated NaCl solution, dried over Na2SO4, and the solvent was removed by vacuum concentration. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 5:1) to give a white solid compound KHL-5 (45.0 g, 88.9%).

[0064] Compound KHL-6: A solution of compound KHL-5 (45.0 g, 128.6 mmol) in tetrahydrofuran (500 mL) was added to adjust the pH to 3 with 1 N HCl solution. After stirring at room temperature for 12 hours, saturated NaCl (1 L) was added, and the mixture was extracted with ethyl acetate (1 L * 3). The organic phases were combined and concentrated to remove the solvent. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 5:1) to give a white solid compound KHL-6 (38.0 g, 97.2%).

[0065] Compound KHL-7: An n-BuLi solution (1.6 M, 164.5 mmol, 102.8 mL) was added to a tetrahydrofuran (200 mL) solution of trimethylsilylacetylene (16.2 g, 164.5 mmol). After stirring at 0 °C for 2 hours, the reaction mixture was cooled to -78 °C, and a tetrahydrofuran (50 mL) solution of compound KHL-6 (10.0 g, 32.9 mmol) was added, followed by stirring for another 2 hours. The reaction mixture was heated to 0 °C, poured into a saturated NH4Cl solution, and extracted with ethyl acetate (500 mL * 3). The organic phases were combined, concentrated to remove the solvent, and the residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 5:1) to give a white solid compound KHL-7 (7.0 g, 52.9%). Compound KHL-8: PCC (62.0 g, 288.6 mmol) was added in portions to a 1 L dichloromethane solution of compound KHL-7 (58.0 g, 144.3 mmol) at 0 °C. After stirring at room temperature for 3 hours, the mixture was filtered and washed with dichloromethane. The organic phase was washed successively with water in 10% Na₂S₂O₃ solution, dried over anhydrous Na₂SO₄, concentrated to remove the solvent, and the residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 10:1) to give a white solid (47.0 g). The solid was then slurried with a petroleum ether / ethyl acetate = 50:1 solvent to give a white solid compound KHL-8 (31.0 g, 53.7%).

[0066] Compound KHL: At 0 °C, TBAF solution (1.0 M, 77.5 mmol, 77.5 mL) was added dropwise to a tetrahydrofuran (500 mL) solution of compound KHL-8 (31.0 g, 77.5 mmol), and the reaction was stirred for 1 hour. The reaction mixture was then added to 1 N HCl solution (77.5 mL) and saturated saline solution (1 L), and extracted with ethyl acetate (1 L * 3). The organic layers were combined, concentrated to remove the solvent, and the residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 5:1) to give a white solid compound L (24.5 g, 96.4%).

[0067] 1 H NMR (400MHz, CDCl3) δ2.55(t,J=8.7Hz,1H),2.50(s,1H),2.17(d,J=12.2Hz,2H),2.13(s,3H),2.10 –1.98(m,2H),1.90(t,J=12.9Hz,2H),1.78–1.38(m,12H),1.33–1.01(m,6H),0.63(s,3H).Mass:m / z 370[M+MeCN+1] +

[0068] Example 2: Synthesis of compound KHL-0021

[0069]

[0070] Compound KHL (100 mg, 0.30 mmol) was mixed with 4'-bromo-2,2,2-trifluoroacetophenone (77 mg, 0.30 mmol), Et3N (0.4 mL), (PPh3)2PdCl2 (4 mg), and CuI (4 mg), followed by 5 mL of dichloromethane. The mixture was reacted at room temperature for 6 hours under nitrogen protection. TLC monitoring showed that the reaction was complete. 20 mL of water was added to the reaction mixture, and the mixture was extracted with dichloromethane (2 x 25 mL). The combined organic phases were washed with saturated sodium chloride water, dried over anhydrous sodium sulfate, and the solvent was removed by vacuum distillation. The mixture was purified using a thick preparative plate (acetone / petroleum ether = 1:2 development) to give a waxy solid (43 mg, 28.7%).

[0071] 1 H NMR(400MHz,Chloroform-d)δ8.01(dt,2H),7.61–7.55(m,2H),2.55(t,1H),2.22–2.14(m,2H),2.12(s,3 H),2.07–1.92(m,3H),1.88–1.80(m,1H),1.66(m,8H),1.51–1.40(m,3H),1.33–1.03(m,6H),0.63(s,3H).

[0072] Example 3: Synthesis of compound KHL-0024

[0073]

[0074] Compound KHL (100 mg, 0.30 mmol) was mixed with 2-bromo-5-methylthiophene (54 mg, 0.30 mmol), Et3N (92 mg, 0.91 mmol), (PPh3)2PdCl2 (11 mg), and CuI (3 mg), and 5 mL of acetonitrile was added. The mixture was heated at 60 °C for 8 hours under nitrogen protection. TLC monitoring showed that the reaction was complete. 20 mL of water was added to the reaction solution, and the mixture was extracted with ethyl acetate (2 x 25 mL). The combined organic phases were washed with saturated sodium chloride water, dried over anhydrous sodium sulfate, and the solvent was removed by vacuum distillation. The mixture was purified by thick preparative agar (ethyl acetate / petroleum ether = 1:3 development) to give a white solid (50 mg, 38.8%).

[0075] 1H NMR(400MHz,Chloroform-d)δ6.98(d,1H),6.61(dd,1H),2.54(t,1H),2.45(s,3H),2.20–2.13(m,1H),2.12( s,3H),2.07–1.87(m,3H),1.83–1.77(m,1H),1.77–1.49(m,9H),1.43(m,3H),1.34–1.01(m,6H),0.62(s,3H).

[0076] Example 4: Synthesis of compound KHL-0025

[0077]

[0078] Compound KHL (150 mg, 0.46 mmol) was mixed with 1-(4-bromo-2-thiophene) ethyl ketone (94 mg, 0.46 mmol), Et3N (139 mg, 1.38 mmol), (PPh3)2PdCl2 (32 mg), and CuI (9 mg), and 5 mL of acetonitrile was added. The mixture was heated at 50 °C for 8 hours under nitrogen protection. TLC monitoring showed that the reaction was complete. 20 mL of water was added to the reaction solution, and the mixture was extracted with ethyl acetate (2 x 25 mL). The combined organic phases were washed with saturated sodium chloride water, dried over anhydrous sodium sulfate, and the solvent was removed by vacuum distillation. The mixture was purified by thick preparative agar (ethyl acetate / petroleum ether = 1:3 development) to give a white solid (104 mg, 50.5%).

[0079] 1 H NMR(400MHz,Chloroform-d)δ7.72–7.62(m,2H),2.55(m,4H),2.20–2.14(m,1H),2.12(s,3H),2.0 9–1.90(m,3H),1.84–1.78(m,1H),1.77–1.51(m,9H),1.45(m,3H),1.33–1.03(m,6H),0.62(s,3H).

[0080] Synthesis of compound KHL-0026 in Example 5

[0081]

[0082] Compound KHL (150 mg, 0.46 mmol) was added to 2-bromo-4-methylthiophene (89 mg, 0.50 mmol), Et3N (139 mg, 1.37 mmol), (PPh3)2PdCl2 (16 mg), and CuI (5 mg). 5 mL of acetonitrile was added, and the mixture was heated at 60 °C for 6 hours under nitrogen protection. The reaction solution changed from a pale yellow and clear liquid to a reddish-brown solution. TLC monitoring showed that the reaction was complete. 30 mL of water was added to the reaction solution, and the mixture was extracted with ethyl acetate (2 x 35 mL). The combined organic phases were washed with saturated sodium chloride water, dried over anhydrous sodium sulfate, and the solvent was removed by vacuum distillation. The solution was purified by rapid silica gel column chromatography (eluting with dichloromethane) to give a white, foamy solid (105 mg). Further purification using a thick preparative plate yielded a white, foamy solid (50 mg, 25.6%).

[0083] 1 H NMR(400MHz,Chloroform-d)δ7.00(d,1H),6.81(t,1H),2.54(t,1H),2.21(s,3H),2.19–2.14(m,2H),2 .12(s,3H),2.08–1.88(m,3H),1.83–1.77(m,1H),1.77–1.38(m,11H),1.33–1.01(m,6H),0.62(s,3H).

[0084] Synthesis of compound KHL-0027 in Example 6

[0085] Compound KHL (150 mg, 0.46 mmol) was mixed with 4-bromo-2-carboxythiophene (87 mg, 0.46 mmol), Et3N (373 mg, 3.68 mmol), (PPh3)2PdCl2 (6 mg), and CuI (5 mg). 5 mL of acetonitrile was added, and the mixture was heated at 70 °C for 3 hours under nitrogen protection. The reaction solution changed from a pale yellow, clear liquid to a black, turbid solution. TLC monitoring showed that the reaction was complete. 20 mL of water was added to the reaction solution, and the mixture was extracted with ethyl acetate (2 x 25 mL). The combined organic phases were washed with saturated sodium chloride water, dried over anhydrous sodium sulfate, and the solvent was removed under reduced pressure to obtain 448 mg of a pale yellow crude solid. The crude solid was purified by rapid silica gel column chromatography (eluting with dichloromethane / ethyl acetate = 30:1) to give 152 mg (75.44%).

[0086] 1H NMR(400MHz,Chloroform-d)δ9.88(d,1H),7.78(t,1H),7.75(d,1H),2.54(t,1H),2.26–2.14(m,2H),2.1 2(s,3H),2.07–1.90(m,3H),1.82(m,1H),1.76–1.51(m,8H),1.45(m,3H),1.34–1.02(m,6H),0.62(s,3H).

[0087] Synthesis of compound KHL-0028 in Example 7

[0088]

[0089] Compound KHL (164 mg, 0.50 mmol) was mixed with 4-bromo-2-trifluoroacetylthiophene (129 mg, 0.50 mmol), Et3N (405 mg, 4.00 mmol), (PPh3)2PdCl2 (7 mg), and CuI (6 mg), and 5 mL of acetonitrile was added. The mixture was heated at 70 °C for 6 hours under nitrogen protection. TLC monitoring showed that the reaction was complete. 20 mL of water was added to the reaction solution, and the mixture was extracted with ethyl acetate (2 x 25 mL). The combined organic phases were washed with saturated sodium chloride water, dried over anhydrous sodium sulfate, and the solvent was removed by vacuum distillation. The mixture was purified by thick preparative agar (acetone / petroleum ether = 1:2 development) to give a white solid (51 mg, 20.2%).

[0090] 1 H NMR(400MHz,Chloroform-d)δ7.91(q,1H),7.90(d,1H),2.54(t,1H),2.22–2.14(m,1H),2.12(s ,3H),2.09–1.91(m,3H),1.86–1.79(m,1H),1.78–1.38(m,12H),1.32–1.04(m,6H),0.63(s,3H).

[0091] Synthesis of compound KHL-0029 in Example 8

[0092]

[0093] Compound KHL (164 mg, 0.50 mmol) was added to 4-bromothiazole (82 mg, 0.50 mmol), Et3N (405 mg, 4.00 mmol), (PPh3)2PdCl2 (7 mg), and CuI (6 mg), followed by 5 mL of acetonitrile. The mixture was heated at 60 °C for 6 hours under nitrogen protection. The reaction solution, initially pale yellow and clear, turned into a brown solution. TLC monitoring showed that the reaction was complete. 20 mL of water was added to the reaction solution, and the mixture was extracted with ethyl acetate (2 x 25 mL). The combined organic phases were washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and the solvent was removed under reduced pressure. The solution was purified using a preparative plate (acetone / petroleum ether = 1:2 development) to give a white solid (54 mg, 26.3%).

[0094] 1 ¹H NMR (400MHz, Chloroform-d) δ 8.78 (d, 1H), 7.50 (d, 1H), 2.54 (t, 1H), 2.20–2.14 (m, 1H), 2.12 (s, 3H), 2.06–1.82 (m, 4H), 1.80–1.37 (m, 10H), 1.34–1.00 (m, 8H), 0.62 (s, 3H). Example 9: Synthesis of compound KHL-0032

[0095]

[0096] Compound KHL (150 mg, 0.46 mmol) was mixed with 2-bromo-5-chlorothiophene (90 mg, 0.46 mmol), Et3N (139 mg, 1.38 mmol), (PPh3)2PdCl2 (32 mg), and CuI (9 mg), followed by 5 mL of acetonitrile. The mixture was heated at 50 °C for 8 hours under nitrogen protection. TLC monitoring showed that the reaction was complete. 20 mL of water was added to the reaction solution, and the mixture was extracted with ethyl acetate (2 x 25 mL). The combined organic phases were washed with saturated sodium chloride water, dried over anhydrous sodium sulfate, and the solvent was removed by vacuum distillation. The mixture was purified by thick preparative agar (ethyl acetate / petroleum ether = 1:3 development) to give a white solid (49 mg, 24.1%).

[0097] 1 H NMR(400MHz,Chloroform-d)δ6.95(d,1H),6.77(d,1H),2.54(t,1H),2.24–2.13(m,2H),2.12(s,3H), 2.07–1.88(m,3H),1.80(m,1H),1.77–1.50(m,8H),1.49–1.39(m,3H),1.32–1.02(m,6H),0.62(s,3H).

[0098] Example 10 Synthesis of compound KHL-0033

[0099] Compound KHL (150 mg, 0.46 mmol) was mixed with 2-bromofuran (67 mg, 0.46 mmol), Et3N (139 mg, 1.38 mmol), (PPh3)2PdCl2 (32 mg), and CuI (9 mg), followed by 5 mL of acetonitrile. The mixture was heated at 50 °C for 8 hours under nitrogen protection. TLC monitoring showed that the reaction was complete. 20 mL of water was added to the reaction solution, and the mixture was extracted with ethyl acetate (2 x 25 mL). The combined organic phases were washed with saturated sodium chloride water, dried over anhydrous sodium sulfate, and the solvent was removed by vacuum distillation. The mixture was purified by thick preparative agar (ethyl acetate / petroleum ether = 1:3 development) to give a white solid (25 mg, 13.9%).

[0100] 1 H NMR(400MHz,Chloroform-d)δ7.37(d,1H),6.57(d,1H),6.38(dd,1H),2.54(t,1H),2.22–2.12(m,2H), 2.12(s,3H),2.06–1.89(m,3H),1.86–1.79(m,1H),1.78–1.39(m,11H),1.35–1.01(m,6H),0.62(s,3H).

[0101] Example 11 Synthesis of compound KHL-0035

[0102] Compound KHL (150 mg, 0.46 mmol) was mixed with 5-bromobenzothiophene (97 mg, 0.46 mmol), Et3N (139 mg, 1.38 mmol), (PPh3)2PdCl2 (32 mg), and CuI (9 mg), followed by 5 mL of acetonitrile. The mixture was heated at 60 °C for 8 hours under nitrogen protection. TLC monitoring showed that the reaction was complete. 20 mL of water was added to the reaction solution, and the mixture was extracted with ethyl acetate (2 x 25 mL). The combined organic phases were washed with saturated sodium chloride water, dried over anhydrous sodium sulfate, and the solvent was removed by vacuum distillation. The mixture was purified by thick preparative agar (ethyl acetate / petroleum ether = 1:3 development) to give a white solid (24 mg, 11.3%).

[0103] 1H NMR(400MHz,Chloroform-d)δ7.94–7.87(m,1H),7.81(d,1H),7.47(d,1H),7.39(dd,1H),7.30(d,1H),2.55(t,1H),2.16(m,2 H),2.12(s,3H),2.07–1.91(m,3H),1.89–1.81(m,1H),1.81–1.52(m,8H),1.52–1.39(m,3H),1.35–1.02(m,6H),0.63(s,3H).

[0104] The comparative compound used in this invention has the following structure:

[0105]

[0106] The specific synthesis process is based on CN105339381B.

[0107] Example 12 In vitro activity assay

[0108] This invention uses a patch-clamp electrophysiological method based on the α1β2γ2 subtype of the recombinant GABA(A) receptor to evaluate the in vitro activity of compounds.

[0109] 12.1 Materials and Methods

[0110] 12.1.1 Cell Culture

[0111] GABA A (α1β2γ2) cell culture

[0112] ●Genetic information:

[0113] α1: GABRA1, NM_000806

[0114] β2: GABRB2, NM_021911

[0115] γ2: GABRG2, NM_198904

[0116] ● Stable expression of GABA A The HEK293 cell line containing the (α1β2γ2) receptor was cultured in DMEM medium containing 10% fetal bovine serum, 800 μg / mL LG418, 200 μg / mL Hygromycin B, and 100 μg / mL Zeocin at 37°C and 5% carbon dioxide.

[0117] ● Cell passage: Remove the old culture medium and wash once with PBS, then add 1 mL of 0.25% Trypsin-EDTA solution.

[0118] Incubate at 37°C for 0.5 min. Once the cells detach from the bottom of the dish, add 5 mL of preheated (37°C) complete culture medium. Gently pipette the cell suspension to separate any aggregated cells. Transfer the cell suspension to a sterile centrifuge tube and centrifuge at 1000 rpm for 5 min to collect the cells. For expansion or maintenance culture, seed the cells in 6 cm cell culture dishes at a density of 2.5 × 10⁶ cells per dish. 5 cells (final volume: 5 mL).

[0119] To maintain the electrophysiological activity of cells, the cell density must not exceed 80%.

[0120] ● Before the experiment, cells were separated using 0.25% Trypsin-EDTA, and 8×10 3 Cells were seeded onto coverslips and cultured in 24-well plates (final volume: 500 μL). After 18 hours, patch-clamp assays were performed. All procedures were performed in accordance with the standard operating procedures for cell culture of Beijing Aisiyipu Biotechnology Co., Ltd.

[0121] 12.2 Electrophysiological Recording

[0122] 12.2.1 Intracellular and extracellular fluid

[0123] ● Extracellular fluid: 140mM NaCl, 5mM CsCl, 2mM CaCl2·2H2O, 1mM MgCl2·6H2O, 5mM MEPES, 10mM D-Glucose, pH=7.4 (NaOH)

[0124] ●Intracellular fluid: 130mM CsCl, 0.1mM CaCl2·2H2O, 2mM MgCl2·6H2O, 1.1mM EGTA, 5mM

[0125] Na 2- ATP, 10mM HEPES, pH=7.2 (CsOH)

[0126] After the intracellular fluid was prepared, it was dispensed into 1 mL tubes and stored at -20°C. Freshly thawed electrode fluid was used daily for the experiment, and all intracellular fluid was used up within three months.

[0127] 12.2.2 Experimental methods refer to

[0128] A capillary glass tube (BF150-86-10, Sutter Instruments) was drawn into a recording electrode using a microelectrode drawing instrument (P97, Sutter Instruments). Under an inverted microscope (IX71, Olympus), the microelectrode manipulator (MP285, Sutter Instruments) was used to bring the recording electrode into contact with the cell, and negative pressure was applied to aspirate and form a GΩ seal. After forming the GΩ seal, rapid capacitance compensation was performed, and then negative pressure was continued to rupture the cell membrane, establishing a whole-cell recording mode. Slow capacitance compensation was then performed, and membrane capacitance and series resistance were recorded. No leakage compensation was applied.

[0129] A coverslip containing cells was placed in the recording bath of an inverted microscope. The working solution of the test sample and the external solution without compounds were sequentially flowed through the recording bath from low to high concentration using gravity perfusion, thus acting on the cells. Liquid exchange was performed using a vacuum pump during recording. Multiple data were collected for each concentration. All electrophysiological experiments were performed at room temperature.

[0130] Whole-cell patch-clamp recording of GABA A1 The voltage stimulation protocol for receptor currents is as follows: after whole-cell sealing, the cell membrane voltage is clamped at -70 mV. GABA is recorded in gap-free mode. A1 Current. After the cells stabilized, 3 μM GABA was administered to stimulate them. After incubation at the tested concentration for 30 s, mixtures of 3 μM GABA and different concentrations of compounds were administered sequentially, with a final administration of 100 μM GABA. The time interval between each administration was 2 min. The experimental data were acquired by an EPC-10 amplifier (HEKA) and stored in PatchMaster (HEKA) software.

[0131] 12.3 Data Quality Standards

[0132] The following criteria are used to determine whether data is acceptable:

[0133] (1) Electrode resistance <5MΩ

[0134] (2) Sealing resistance > 1 GΩ

[0135] (3) Initial connection resistance <15MΩ

[0136] (4) Connection resistor ends <15MΩ

[0137] (5) The current does not show obvious spontaneous decay.

[0138] (6) No significant leakage current was observed at a membrane potential of -70mV.

[0139] 12.4 Data Analysis

[0140] In detecting GABA A1 At that time, the currents of 3 μM GABA and mixtures prepared with different drug concentrations were standardized with the current of 100 μM GABA. Then, the activating rate for each drug concentration was calculated. The mean and standard error were calculated for each concentration. The half-activating concentration for each compound was then calculated using the following equation:

[0141] Y=Bottom+Bottom / (1+10^((LogEC 50 -X)×HillSlope))

[0142] The dose-dependent effect was nonlinearly fitted using the above equations, where EC 50 The half-activation concentration is represented by Hillslope, and the coefficient of performance (EC) is also shown. Curve fitting and EC are also discussed. 50 The calculations were performed using Graphpad 5.0 software.

[0143] 12.5 Results

[0144] Table 1 shows the efficacy of the test sample at two concentrations of 0.1 μM and 1 μM, and the ratio of the peak current generated by the test sample at these two concentrations when acting on the GABA(A) receptor in conjunction with 3 μmol GABA to the peak current generated when 100 μmol GABA is present alone, multiplied by 100%.

[0145] Table 1: Effects of different compounds on GABA (A) Electrophysiological evaluation of channel activating effect

[0146] Compound numbering 0.1μM 1μM KHL-0021 F F KHL-0024 E A KHL-0025 F D KHL-0026 F B KHL-0027 F E KHL-0028 F E KHL-0029 E C KHL-0032 E C KHL-0033 F D KHL-0035 B B Comparative compounds F D

[0147] A: >60%, B: 50%-60% (inclusive), C: 40%-50% (inclusive), D: 30%-40% (inclusive), E: 20%-30% (inclusive), F: ≤20%

[0148] Table 2: Effects of different compounds on GABA (A) Electrophysiological evaluation of channel activating effect

[0149] compound EC50 (nmol) Emax KHL-0026 1601 control compound 1261 84.2％

[0150] Emax is defined as the ratio of the peak current generated when the test sample is present with 3 μmol GABA to the peak current generated when 100 μmol GABA is present alone, multiplied by 100%.

Claims

1. A compound of formula I and a pharmaceutically acceptable salt thereof: R1 is selected from substituted or unsubstituted five-membered aromatic heterocycles, substituted or unsubstituted thiazolyl groups, and substituted or substituted benzothiophene groups; R2 is a hydrogen atom or a methyl group.

2. The compound of formula I according to claim 1 and its pharmaceutically acceptable salt, characterized in that... The substituted or unsubstituted five-membered aromatic heterocycle is selected from monosubstituted ones. or double substitution or tri-substitution Where X is O or S; R3, R4, and R5 are each independently hydrogen, substituted or unsubstituted C1-6 alkyl, halogen, -NO2, -CN, -OR, -N(R)2, -C(=O)R, -C(=O)OR, -OC(=O)R, -OC(=O)OR, -C(=O)N(R)2, -N(R)C(=O)R, -OC(=O)N(R)2, -N(R)C(=O)OR, -N(R)C(=O)N(R)2, -SR, -S(O)R, for example -S(=O)R, -S(=O)2R, -S(=O)2OR, -OS(=O)2R, -S(=O)2N(R)2, -N(R)S(=O)2R, substituted or unsubstituted C2-6 alkenyl, substituted or unsubstituted C2-6 alkynyl.

3. The compound of formula I according to claim 1 and its pharmaceutically acceptable salt, characterized in that... The substituted or unsubstituted thiazolyl group is selected from monosubstituted groups. or double substitution R3 and R 45 Each is independently hydrogen, substituted or unsubstituted C1-6 alkyl, halogen, -NO2, -CN, -OR, -N(R)2, -C(=O)R, -C(=O)OR, -OC(=O)R, -OC(=O)OR, -C(=O)N(R)2, -N(R)C(=O)R, -OC(=O)N(R)2, -N(R)C(=O)OR, -N(R)C(=O)N(R)2, -SR, -S(O)R, for example -S(=O)R, -S(=O)2R, -S(=O)2OR, -OS(=O)2R, -S(=O)2N(R)2, -N(R)S(=O)2R, substituted or unsubstituted C2-6 alkenyl, substituted or unsubstituted C2-6 alkynyl.

4. The compound of formula I according to claim 1 and its pharmaceutically acceptable salt, characterized in that... The benzene-substituted or unsubstituted thiazolyl group is selected from... in R3 is hydrogen, substituted or unsubstituted C1-6 alkyl, halogen, -NO2, -CN, -OR, -N(R)2, -C(=O)R, -C(=O)OR, -OC(=O)R, -OC(=O)OR, -C(=O)N(R)2, -N(R)C(=O)R, -OC(=O)N(R)2, -N(R)C(=O)OR, -N(R)C(=O)N(R)2, -SR, -S(O)R, for example -S(=O)R, -S(=O)2R, -S(=O)2OR, -OS(=O)2R, -S(=O)2N(R)2, -N(R)S(=O)2R, substituted or unsubstituted C2-6 alkenyl, substituted or unsubstituted C2-6 alkynyl.

5. The compound of formula I according to any one of claims 1-4 and its pharmaceutically acceptable salt, characterized in that:

6. A pharmaceutical composition comprising a compound of any one of claims 1-5 or a pharmaceutically acceptable salt thereof, and a pharmaceutically usable excipient.

7. The use of the compound of formula I as described in any one of claims 1-5 and its pharmaceutically acceptable salt, and the use of the pharmaceutical composition of claim 6 in the preparation of a medicament for the prevention or treatment of nervous system diseases.

8. The use according to claim 7, characterized in that... The neurological disorders are selected from: sleep disorders, mood disorders, schizophrenia spectrum disorders, spastic disorders, memory disorders and / or cognitive disorders, motor disorders, personality disorders, autism spectrum disorders, pain, traumatic brain injury, vascular diseases, substance abuse disorders and / or truncation syndrome, or tinnitus.

9. The use according to claim 8, characterized in that... The mood disorder is depression; preferably major depression or postpartum depression.

10. The pharmaceutical composition according to claim 6, characterized in that... The pharmaceutical composition can be administered orally, subcutaneously, intravenously, or intramuscularly.

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