Steroidal compounds, uses thereof and preparation methods therefor

Abstract

The present invention relates to steroid compounds, uses thereof and preparation methods thereof; it is expected that such compounds can effectively treat mental and neurological diseases and have good activity efficacy, pharmacokinetic (PK) properties, oral bioavailability, stability, safety, clearance rate and / or metabolic properties, etc.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceuticals, and specifically to steroid compounds, uses thereof and preparation methods therefor. Technical Background

[0002] Neuropsychiatric diseases, including anxiety disorders, depression, schizophrenia, etc., rank first in the total disease burden in China, accounting for approximately 20% of the total disease burden (Wang Juncheng et al., Chinese Health Service Management, 2009(5): 348 - 350). With the continuous development of the social modernization process, the accelerating work rhythm and increasing life pressure of people have led to a significant increase in the number of patients with various neuropsychiatric diseases, and the symptom progression has accelerated significantly. Therefore, it is more urgent to develop, research and produce drugs for treating neuropsychiatric diseases. At the same time, clinical data show that many patients are troubled by multiple neuropsychiatric diseases simultaneously. The high comorbidity rate of neuropsychiatric diseases poses a greater challenge to clinical treatment.

[0003] In animals, GABA (gamma - aminobutyric acid) is only present in nerve tissues. Immunological studies have shown that the region with the highest concentration is the substantia nigra in the brain. GABA is an important inhibitory neurotransmitter that has been relatively well - studied at present. It participates in various metabolic activities and has high physiological activity. The GABA system is the main inhibitory signaling pathway in the brain and central nervous system, and is of great significance for regulating the function of the central nervous system. The GABA(A) receptor (GABAAR) is an ionotropic receptor and a ligand - gated ion channel. Its endogenous ligand is gamma - aminobutyric acid (GABA), the main inhibitory neurotransmitter in the central nervous system. After the activation of the GABA(A) receptor, it selectively allows Cl - to pass through its pore. Cl - flows out of the neuron cell when the internal voltage is less than the resting potential, and flows into the neuron cell when the internal voltage is greater than the resting potential (i.e., - 75 mV). This can successfully reduce the chance of action potential occurrence and has an inhibitory effect on neurotransmission. Anxiety disorders 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 results have confirmed that the GABAergic system and the GABA(A) receptor play important roles in the etiopathology of these three diseases, indicating that a pathophysiological process related to the GABAergic system may be one of the common determinants of these 3 diseases. The GABAergic system and the GABA(A) receptor have been confirmed to participate in the pathological processes of anxiety, depression and schizophrenia at the molecular level, pre - clinical and clinical levels, and the GABA(A) receptor has long been an important drug target for the treatment of these diseases.

[0004] Clinical trials have confirmed that, in addition to binding to GABA itself to form a receptor complex (GRC) that alters brain excitability, the GABA(A) receptor can also bind to small molecule compounds with some specific structures, such as barbiturate drugs (trade name Seconal) and benzodiazepine drugs (trade name diazepam), etc. These drugs bind to specific allosteric sites on the GABA(A) receptor respectively to produce their therapeutic effects. In addition, there are also studies showing that there are unique sites for steroid (steroidal) compounds on the GABA(A) receptor (Lan, N.C. et al., Neurochem.Res. 16:347-356 (1991)).

[0005] Neuroactive steroids can occur endogenously. The most potent endogenous neuroactive steroids are 3α-hydroxy-5-reduced pregnane-20-one and 3α-21-dihydroxy-5-reduced pregnane-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.L. et al, J Pharmacol. Exp. Ther. 241:346-353 (1987)). 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.W. and McEwen, B.S., et al, 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. It has been demonstrated in many literatures that the levels of progesterone and its metabolites decrease before the onset of menstruation. Some physical symptoms that recur monthly before the onset of menstruation have also been demonstrated in many literatures. These symptoms that have become associated with premenstrual syndrome (PMS) include stress (tension), anxiety, and migraine (Dalton, K., Premenstrual Syndrome and Progesterone Therapy, 2nd edition, Chicago Year book, Chicago (1984)). Subjects with PMS have symptoms that recur monthly before menstruation and disappear after menstruation.

[0006] Additional studies have also found that a decrease in progesterone is associated with an increase in the seizure frequency of female epilepsy patients, i.e., catamenial epilepsy (Laidlaw, J., Lancet, 1235 - 1237 (1956)). A more direct association has been observed for a decrease in progesterone metabolites (Rosciszewska et al., J. Neurol. Neurosurg. Psych.. 49:47 - 51 (1986)). Additionally, for patients with primary generalized petit mal epilepsy, the disease incidence has been related to the incidence of premenstrual symptoms (Backstrom, T. et al, J. Psychosom. Obstet. Gynaecol. 2:8 - 20 (1983)). The steroid deoxycorticosterone has been found to be effective in treating epilepsy patients related to the menstrual cycle (Aird, R. B. and Gordan, G., J. Amer. Med. Soc. 145:715 - 719 (1951)).

[0007] In addition to the above - mentioned conditions, postpartum depression (PND) is also associated with low progesterone levels. Immediately after childbirth, progesterone levels drop sharply, leading to the onset of PND. The symptoms of PND range from mild depression to psychosis requiring hospitalization. PND is also associated with severe anxiety and irritability. Depression related to PND cannot be treated with classical antidepressants, and the probability of developing PMS in women experiencing PND is increased (Dalton, K., Premenstrual Syndrome and Progesterone Therapy, 2nd edition, Chicago Yearbook, Chicago (1984)). These observations more or less imply a key role of progesterone and deoxycorticosterone, especially their metabolites, in regulating the excitatory state of the brain, and their related clinical indications are shown as catamenial epilepsy, PMS, and PND.

[0008] Many studies are based on the GABA(A) receptor in order to obtain drugs that can effectively treat related diseases. CN103958540A, CN10533928A, etc. disclose a series of neurosteroid compounds for treating psychoneurological diseases. GABA(A): a subtype of the γ - aminobutyric acid receptor (GABAR). There are 3 receptor subtypes of GABAR, including GABA(A), GABA(B), and GABA(C). The GABA(A) receptor is a chloride channel.

[0009] GABA(A) is composed of five subunits (polypeptide chains) that combine to form a pentagonal hetero-oligomeric structure. Among the five subunits, two pairs are the same, so a specific GABA(A) receptor contains three types of subunits, commonly α, β, and γ. Each subunit has several different subtypes. For example, α has α1, α2, α3, α4, α5, α6; β has β1, β2, β3; γ has γ1, γ2, γ3, γ4. In addition, there are also subunits such as δ, ε, ρ1-3, θ, π, etc., which can form pentamers with α and β.

[0010] According to statistics, there are currently 23 different subunit combinations of GABA(A) receptors. The specific combination form of ligands has a certain relationship with the distribution area in brain neurons. For example, GABA(A) receptors containing the γ subunit are mainly distributed within synapses, and GABA(A) receptors containing the δ subunit are mainly distributed extrasynaptic (Activation of GABA(A) Receptors: Views from Outside the Synaptic Cleft, Neuron. vol 56, 2007, 763-770); according to existing research (Endogenous neurosteroids regulate GABAA receptors through two discrete transmembrane sites: Nature. Vol 444. 23 November. 2006. 486-489), endogenous steroid compounds such as tetrahydrodeoxycorticosterone (THDOC) can directly activate the receptor subtype α1β2γ2 within synapses at high concentrations (micromolar levels) (hereinafter referred to as GABA A1 ), and at lower concentrations (submicromolar levels), it can enhance the effect amplitude of GABA on α1β2γ2.

[0011]

[0012] However, there are no reports of direct activation of extrasynaptic receptor subtypes (such as α4β3δ, hereinafter referred to as GABA A4 ) by steroid compounds. SUMMARY OF THE INVENTION

[0013] One of the objectives of the present invention is to provide a steroid compound with better efficacy, which can more effectively treat mental and neurological diseases. The compounds of the present invention are expected to provide good activity potency, pharmacokinetic (PK) properties, oral bioavailability, formulatability, stability, safety, clearance rate, and / or metabolic properties, and reduce side effects such as anesthesia and sedation.

[0014] For the above purposes, the present invention provides a compound of formula I and its pharmaceutically acceptable salts:

[0015]

[0016] Wherein R1 is hydrogen, a substituted or unsubstituted C1-6 alkyl group, a substituted or unsubstituted C1-6 alkoxy group, a substituted or unsubstituted C2-6 alkenyl group, a substituted or unsubstituted C2-6 alkynyl group, or a substituted or unsubstituted C3-6 carbocyclic group;

[0017] R2 is hydrogen, halogen, a substituted or unsubstituted C1-6 alkyl group, a substituted or unsubstituted C1-6 alkoxy group, a substituted or unsubstituted

[0018] C2-6 alkenyl group, a substituted or unsubstituted C2-6 alkynyl group, a substituted or unsubstituted C3-6 carbocyclic group;

[0019] R3 is hydrogen, an unsubstituted C1-6 alkyl group or -CH2OR a , wherein R a is hydrogen, a substituted or unsubstituted C1-6 alkyl group, a substituted or unsubstituted C1-6 alkoxy group, a substituted or unsubstituted C2-6 alkenyl group, a substituted or unsubstituted C2-6 alkynyl group, a substituted

[0020] or unsubstituted C3-6 carbocyclic group;

[0021] L is -C(Rb)(Rb)-, each Rb is independently hydrogen or a C1-C6 alkyl group, n is an integer from 0 to 3,

[0022] R4 is halogen or a substituted or unsubstituted heteroaryl group or heterocyclic group.

[0023] For a further preferred compound, R1 is hydrogen, a substituted or unsubstituted C1-6 alkyl group; R2 is hydrogen, a substituted or unsubstituted C1-6 alkyl group; R3 is hydrogen, an unsubstituted C1-6 alkyl group; Rb is hydrogen, n is an integer from 1 to 2; R4 is a heteroaryl group and is optionally substituted by cyano, nitro, hydroxy, halo, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, -C(O)Rd, -C(O)N(Re)(Rf), -C(O)O(Rd), -N(Re)(Rf), -OC(O)N(Re)(Rf), -OC(O)O(Rd), -OC(O)Rd, -S(O) 0-2 Rd, -S(O) 0-2 ORd or -S(O) 0-2 N(Re)(Rf); each Rd is hydrogen or a C1-C6 alkyl group; each Re and Rf are independently hydrogen, a C1-C6 alkyl group, a carbocyclic group, a heterocyclic group, an aryl group, a heteroaryl group.

[0024] Further preferred compounds, R1 is hydrogen or methyl; R2 is hydrogen; R3 is hydrogen; L is CH2; R4 is a monocyclic or bicyclic heteroaryl or a monocyclic or bicyclic heterocyclic group, wherein the heteroatom is oxygen or nitrogen or sulfur, and the bicyclic is selected from spiro or fused rings.

[0025] Further preferred compounds, wherein R4 is a 5- or 6-membered heteroaryl containing 2-4 nitrogen atoms, and is optionally substituted by cyano, nitro, hydroxyl, halogen, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl.

[0026] Or as shown in the compound of formula II:

[0027]

[0028]

[0029] Wherein, R1 is hydrogen, C1-3 alkyl substituted or unsubstituted by halogen; R2 is hydrogen, halogen, C1-6 alkyl or C1-6 alkoxy substituted or unsubstituted by halogen; X is CH2, N, O or S; R5 is selected from the following groups:

[0030]

[0031] Wherein R6 is H, halogen, CN, CF3, NO2, C1-6 alkyl or C1-6 alkoxy substituted or unsubstituted by halogen.

[0032] Or, the compounds of the present invention are preferably selected from:

[0033]

[0034]

[0035]

[0036] The present invention also provides a pharmaceutical composition, which comprises any one of the above-mentioned compounds or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

[0037] The compounds described herein can act as GABA modulators, for example, affecting the GABA(A) receptor in a positive or negative manner. As a modulator of the excitability of the central nervous system (CNS), when mediated by its ability to regulate the GABA(A) receptor, such compounds are expected to have CNS activity.

[0038] The present invention further provides the use of the above-mentioned compound or pharmaceutical composition in the preparation of a medicament for preventing or treating a nervous system disease; wherein the nervous system disease is preferably selected from: sleep disorder, mood disorder, schizophrenia spectrum disorder, spastic disorder, memory disorder and / or cognitive disorder, movement disorder, personality disorder, autism spectrum disorder, pain, traumatic brain injury, vascular disease, substance use disorder and / or truncation syndrome or tinnitus; the mood disorder is depression; wherein the depression is disruptive mood dysregulation disorder, major depressive disorder, persistent depressive disorder, premenstrual dysphoric disorder, substance- or drug-induced disorder, disorder due to another medical condition, other specified depressive disorder and unspecified depressive disorder, preferably mild depression, moderate depression, severe depression or postpartum depression. Further preferably severe depression or postpartum depression. And the above-mentioned compound or pharmaceutical composition can be administered by means such as oral, subcutaneous, intravenous or intramuscular administration.

[0039] Definitions

[0040] Chemical Definitions

[0041] The compounds described herein may include one or more asymmetric centers and may thus exist in various isomeric forms, for example, enantiomeric and / or diastereomeric forms. For example, the compounds described herein may be individual enantiomers, diastereomers or geometric isomers, or may be in the form of a mixture of stereoisomers, including racemic mixtures and mixtures enriched in one or more stereoisomers. The isomers can be separated from the mixture 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 the preferred isomers can be prepared by asymmetric synthesis.

[0042] The following terms are intended to have the meanings provided below and are useful in understanding the specification and intended scope of the present invention. When describing the present invention, it may include compounds, pharmaceutical compositions containing the compounds, and test methods for the compounds and compositions. The term definitions involved in the present invention can be referred to the following description, and any part defined below can be substituted by many substituents, and the corresponding definitions are within the scope listed below, including such substituted parts. Unless otherwise stated, the term "substituted" is defined as follows.

[0043] "Alkyl" refers to a straight-chain or branched-chain saturated hydrocarbon group having 1 to 20 carbon atoms ("C1-20 alkyl"). In some embodiments, the alkyl has 1 to 12 carbon atoms ("C1-12 alkyl"). In some embodiments, the alkyl has 1 to 10 carbon atoms ("C1-10 alkyl"). In some embodiments, the alkyl has 1 to 9 carbon atoms ("C1-9 alkyl"). In some embodiments, the alkyl has 1 to 8 carbon atoms ("C1-8 alkyl"). In some embodiments, the alkyl has 1 to 7 carbon atoms ("C1-7 alkyl"). In some embodiments, the alkyl has 1 to 6 carbon atoms ("C1-6 alkyl", which also refers to "lower alkyl" herein). In some embodiments, the alkyl has 1 to 5 carbon atoms ("C1-5 alkyl"). In some embodiments, the alkyl has 1 to 4 carbon atoms ("C1-4 alkyl"). In some embodiments, the alkyl has 1 to 3 carbon atoms ("C1-3 alkyl"). In some embodiments, the alkyl has 1 to 2 carbon atoms ("C1-2 alkyl"). In some embodiments, the alkyl has 1 carbon atom ("C1 alkyl"). In some embodiments, the alkyl has 2 to 6 carbon atoms ("C2-6 alkyl"). Examples of C1-6 alkyl 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 include n-heptyl (C7), n-octyl (C8), and so on. Unless otherwise specified, each alkyl is independently optionally substituted, i.e., unsubstituted ("unsubstituted alkyl") or substituted by one or more substituents ("substituted alkyl"); for example, 1 to 5 substituents, 1 to 3 substituents, or 1 substituent. In some embodiments, the alkyl is unsubstituted C1-10 alkyl (e.g., -CH3). In some embodiments, the alkyl is substituted C1-10 alkyl.

[0044] "Alkenyl" refers to a straight-chain or branched hydrocarbon group 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) ("C2-20 alkenyl"). In some embodiments, the alkenyl does not contain any triple bonds. In some embodiments, the alkenyl has 2 to 10 carbon atoms ("C2-10 alkenyl"). In some embodiments, the alkenyl has 2 to 9 carbon atoms ("C2-9 alkenyl"). In some embodiments, the alkenyl has 2 to 8 carbon atoms ("C2-8 alkenyl"). In some embodiments, the alkenyl has 2 to 7 carbon atoms ("C2-7 alkenyl"). In some embodiments, the alkenyl has 2 to 6 carbon atoms ("C2-6 alkenyl"). In some embodiments, the alkenyl has 2 to 5 carbon atoms ("C2-5 alkenyl"). In some embodiments, the alkenyl has 2 to 4 carbon atoms ("C2-4 alkenyl"). In some embodiments, the alkenyl has 2 to 3 carbon atoms ("C2-3 alkenyl"). In some embodiments, the alkenyl has 2 carbon atoms ("C2 alkenyl"). One or more carbon-carbon double bonds can be internal (e.g., in 2-butenyl) or terminal (e.g., in 1-butenyl). Examples of C2-4 alkenyls include: vinyl (C2), 1-propenyl (C3), 2-propenyl (C3), 1-butenyl (C4), 2-butenyl (C4), butadienyl (C4), etc. Examples of C2-6 alkenyls include: the above C2-4 alkenyls, as well as pentenyl (C5), pentadienyl (C5), hexenyl (C6), etc. Other examples of alkenyls include: heptenyl (C7), octenyl (C8), octatrieneyl (C8), etc. Unless otherwise specified, each alkenyl is independently optionally substituted, i.e., unsubstituted ("unsubstituted alkenyl") or substituted with one or more substituents ("substituted alkenyl"); e.g., 1 to 5 substituents, 1 to 3 substituents, or 1 substituent. In some embodiments, the alkenyl is an unsubstituted C2-10 alkenyl. In some embodiments, the alkenyl is a substituted C2-10 alkenyl.

[0045] "Alkynyl" refers to a straight-chain or branched hydrocarbon group ("C2-20 alkynyl") having from 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 alkynyl does not contain any double bonds. In some embodiments, the alkynyl has from 2 to 10 carbon atoms ("C2-10 alkynyl"). In some embodiments, the alkynyl has from 2 to 9 carbon atoms ("C2-9 alkynyl"). In some embodiments, the alkynyl has from 2 to 8 carbon atoms ("C2-8 alkynyl"). In some embodiments, the alkynyl has from 2 to 7 carbon atoms ("C2-7 alkynyl"). In some embodiments, the alkynyl has from 2 to 6 carbon atoms ("C2-6 alkynyl"). In some embodiments, the alkynyl has from 2 to 5 carbon atoms ("C2-5 alkynyl"). In some embodiments, the alkynyl has from 2 to 4 carbon atoms ("C2-4 alkynyl"). In some embodiments, the alkynyl has from 2 to 3 carbon atoms ("C2-3 alkynyl"). In some embodiments, the alkynyl has 2 carbon atoms ("C2 alkynyl"). One or more carbon triple bonds can be internal (e.g., in 2-butynyl) or terminal (e.g., in 1-butynyl). Examples of C2-4 alkynyl include, but are not limited to: ethynyl (C2), 1-propynyl (C3), 2-propynyl (C3), 1-butynyl (C4), 2-butynyl (C4), and the like. Examples of C2-6 alkynyl include: the above C2-4 alkynyl, and pentynyl (C5), hexynyl (C6), and the like. Other examples of alkynyl include heptynyl (C7), octynyl (C8), and the like. Unless otherwise specified, each alkynyl is independently optionally substituted, i.e., unsubstituted ("unsubstituted alkynyl") or substituted with one or more substituents ("substituted alkynyl"); e.g., 1 to 5 substituents, 1 to 3 substituents, or 1 substituent. In some embodiments, the alkynyl is an unsubstituted C2-10 alkynyl. In some embodiments, the alkynyl is a substituted C2-10 alkynyl.

[0046] "Aryl" refers to a group ("C6-14 aryl") having a monocyclic or polycyclic (e.g., bicyclic or tricyclic) 4n+2 aromatic ring system (e.g., having 6, 10, or 14 π electrons shared in a cyclic arrangement) with 6-14 ring carbon atoms and zero heteroatoms in an aromatic ring system. In some embodiments, the aryl has six ring carbon atoms ("C6 aryl"; e.g., phenyl). In some embodiments, the aryl has ten ring carbon atoms ("C10 aryl"; e.g., naphthyl, e.g., 1-naphthyl and 2-naphthyl). In some embodiments, the aryl has fourteen ring carbon atoms ("C14 aryl"; e.g., anthryl). "Aryl" also includes ring systems in which the above aryl ring is fused to one or more carbocyclic or heterocyclic groups, where the moiety or point of attachment 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 aryls include, but are not limited to, groups derived from: aceanthrylene, acenaphthylene, acephenanthrylene, anthracene, azulene, benzene, coronene, fluoranthene, fluorene, hexacene, indane, indene, naphthalene, octacene, octaphene, ovalene, penta-2,4-diene, pentacene, pentaphene, pentaphenylene, perylene, phenalene, phenanthrene, picene, heptacene, pyrene, picene, rubicene, benzophenanthrene, and terphenyl. Specifically, aryl includes phenyl, naphthyl, indenyl, and tetrahydronaphthyl. Unless otherwise specified, each aryl is independently optionally substituted, i.e., unsubstituted ("unsubstituted aryl") or substituted with one or more substituents ("substituted aryl"). In some embodiments, the aryl is an unsubstituted C6-14 aryl. In some embodiments, the aryl is a substituted C6-14 aryl. In some embodiments, the aryl is substituted with one or more groups selected from halogen, C1-C8 alkyl, C1-C8 haloalkyl, cyano, hydroxy, C1-C8 alkoxy, and amino.

[0047] "Heteroaryl" refers to a 5- to 10-membered monocyclic or bicyclic 4n+2 aromatic ring system having ring carbon atoms and 1 to 4 ring heteroatoms in an aromatic ring system (e.g., having 6 or 10 π electrons shared in a cyclic arrangement), where each heteroatom is independently selected from nitrogen, oxygen, and sulfur ("5-10 membered heteroaryl"). In heteroaryls containing one or more nitrogen atoms, the point of attachment can be a carbon or nitrogen atom, provided that the valence allows. The heteroaryl bicyclic system can include one or more heteroatoms in one or both rings. "Heteroaryl" includes ring systems in which the above-described heteroaryl ring is fused to one or more carbocyclic or heterocyclic groups, where the point of attachment 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 above-described heteroaryl ring is fused to one or more aryl groups, where the point of attachment 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. For bicyclic heteroaryls in which one ring does not contain a heteroatom (e.g., indolyl, quinolinyl, carbazolyl, etc.), the point of attachment can be on either ring, i.e., on the ring with the heteroatom (e.g., 2-indolyl) or on the ring without the heteroatom (e.g., 5-indolyl). In some embodiments, heteroaryl is a 5- to 10-membered aromatic ring system having ring carbon atoms and 1 to 4 ring heteroatoms (present in an aromatic ring system), where each heteroatom is independently selected from nitrogen, oxygen, and sulfur ("5-10 membered heteroaryl"). In some embodiments, heteroaryl is a 5- to 8-membered aromatic ring system having ring carbon atoms and 1 to 4 ring heteroatoms (present in an aromatic ring system), where each heteroatom is independently selected from nitrogen, oxygen, and sulfur ("5-8 membered heteroaryl"). In some embodiments, heteroaryl is a 5- to 6-membered aromatic ring system having ring carbon atoms and 1 to 4 ring heteroatoms (present in an aromatic ring system), where each heteroatom is independently selected from nitrogen, oxygen, and sulfur ("5-6 membered heteroaryl"). In some embodiments, the 5-6 membered heteroaryl has 1 to 3 ring heteroatoms selected from nitrogen, oxygen, and sulfur. In some embodiments, the 5-6 membered heteroaryl has 1 to 2 ring heteroatoms selected from nitrogen, oxygen, and sulfur. In some embodiments, the 5-6 membered heteroaryl has 1 ring heteroatom selected from nitrogen, oxygen, and sulfur. Unless otherwise specified, each heteroaryl is independently optionally substituted, i.e., unsubstituted ("unsubstituted heteroaryl") or substituted with one or more substituents ("substituted heteroaryl"). In some embodiments, heteroaryl is an unsubstituted 5- to 14-membered heteroaryl. In some embodiments, heteroaryl is a substituted 5- to 14-membered heteroaryl. 5-membered heteroaryls containing one heteroatom include, but are not limited to: pyrrolyl, furyl, and thienyl. 5-membered heteroaryls containing two heteroatoms include, but are not limited to: imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, and isothiazolyl. 5-membered heteroaryls containing three heteroatoms include, but are not limited to: triazolyl, oxadiazolyl, and thiadiazolyl.Five-membered heteroaryls containing four heteroatoms include, but are not limited to, tetrazolyl. Six-membered heteroaryls containing one heteroatom include, but are not limited to, pyridyl. Six-membered heteroaryls containing two heteroatoms include, but are not limited to, pyridazinyl, pyrimidinyl, and pyrazinyl. Six-membered heteroaryls containing three or four heteroatoms include, but are not limited to, triazinyl and tetrazinyl, respectively. Seven-membered heteroaryls containing one heteroatom include, but are not limited to, azepinyl, oxepinyl, and thiepinyl. 5,6-Bicyclic heteroaryls include, but are not limited to, indolyl, isoindolyl, indazolyl, benzotriazolyl, benzothienyl, isobenzothienyl, benzofuranyl, isobenzofuranyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl, benzoxadiazolyl, benzothiazolyl, benzisothiazolyl, benzothiadiazolyl, indolizinyl, and purinyl. 6,6-Bicyclic heteroaryls include, but are not limited to, naphthyridinyl, pteridinyl, quinolinyl, isoquinolinyl, cinnolinyl, quinoxalinyl, phthalazinyl, and quinazolinyl.

[0048] "Carbocyclic group" or "carbocycle" refers to a non-aromatic cycloalkyl hydrocarbon group having 3 to 10 ring carbon atoms and zero heteroatoms in a non-aromatic ring system ("C3-10 carbocyclic group"). In some embodiments, the carbocyclic group has 3 to 8 ring carbon atoms ("C3-8 carbocyclic group"). In some embodiments, the carbocyclic group has 3 to 6 ring carbon atoms ("C3-6 carbocyclic group"). In some embodiments, the carbocyclic group has 3 to 6 ring carbon atoms ("C3-6 carbocyclic group"). In some embodiments, the carbocyclic group has 5 to 10 ring carbon atoms ("C5-10 carbocyclic group"). Exemplary C3-6 carbocyclic groups include, but are not limited to: cyclopropyl (C3), cyclopropenyl (C3), cyclobutyl (C4), cyclobutenyl (C4), cyclopentyl (C5), cyclopentenyl (C5), cyclohexyl (C6), cyclohexenyl (C6), cyclohexadienyl (C6), and the like. Exemplary C3-8 carbocyclic groups include, but are not limited to: the above C3-6 carbocyclic groups, and cycloheptyl (C7), cycloheptenyl (C7), cycloheptadienyl (C7), cycloheptatrienyl (C7), cyclooctyl (C8), cyclooctenyl (C8), bicyclo[2.2.1]heptyl (C7), bicyclo[2.2.2]octyl (C8), and the like. Exemplary C3-10 carbocyclic groups include, but are not limited to: the above C3-8 carbocyclic groups, and cyclononyl (C9), cyclononenyl (C9), cyclodecyl (C10), cyclodecenyl (C10), octahydro-1H-indenyl (C9), decahydronaphthyl (C10), spiro[4.5]decyl (C10), and the like. As illustrated by the foregoing examples, in some embodiments, the carbocyclic group is a monocyclic ("monocyclic carbocyclic group") or a carbocyclic group containing a fused ring system, a bridged ring system, or a spiro ring system, such as a bicyclic system ("bicyclic carbocyclic group"), and can be a saturated or partially unsaturated carbocyclic group. "Carbocyclic group" also includes a ring system in which the above carbocyclic group ring is fused to one or more aryl or heteroaryl groups, wherein the point of attachment is on the carbocyclic group ring, and in such cases, the number of carbons continues to represent the number of carbons in the carbocyclic system. Unless otherwise specified, each of the carbocyclic groups is independently optionally substituted, i.e., unsubstituted ("unsubstituted carbocyclic group") or substituted with one or more substituents ("substituted carbocyclic group"). 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 group" is a monocyclic saturated carbocyclic group having 3 to 10 ring carbon atoms ("C3-10 cycloalkyl"). In some embodiments, the cycloalkyl has 3 to 8 ring carbon atoms ("C3-8 cycloalkyl"). In some embodiments, the cycloalkyl has 3 to 6 ring carbon atoms ("C3-6 cycloalkyl"). In some embodiments, the cycloalkyl has 5 to 6 ring carbon atoms ("C5-6 cycloalkyl"). In some embodiments, the cycloalkyl has 5 to 10 ring carbon atoms ("C5-10 cycloalkyl").Examples of C5-6 cycloalkyl include cyclopentyl (C5) and cyclohexyl (C6). Examples of C3-6 cycloalkyl include the above C5-6 cycloalkyl, as well as cyclopropyl (C3) and cyclobutyl (C4). Examples of C3-8 cycloalkyl include the above C3-6 cycloalkyl, as well as cycloheptyl (C7) and cyclooctyl (C8). Unless otherwise specified, in each case, the cycloalkyl is independently unsubstituted ("unsubstituted cycloalkyl") or substituted with one or more substituents ("substituted cycloalkyl"). In some embodiments, the cycloalkyl is unsubstituted C3-10 cycloalkyl. In some embodiments, the cycloalkyl is substituted C3-10 cycloalkyl.

[0049] "Heterocyclic group" or "heterocycle" refers to a group of atoms of a 3- to 10-membered non-aromatic ring system having ring carbon atoms and 1 to 4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, sulfur, boron, phosphorus, and silicon ("3- to 10-membered heterocyclic group"). In a heterocyclic group containing one or more nitrogen atoms, the point of attachment can be a carbon or nitrogen atom, provided that the valence allows. The heterocyclic group can be monocyclic ("monocyclic heterocyclic group") or a fused ring system, bridged ring system, or spiro ring system, such as a bicyclic system ("bicyclic heterocyclic group"), and can be a saturated or partially unsaturated heterocyclic group. The bicyclic system of the heterocyclic group can include one or more heteroatoms in one or both rings. "Heterocyclic group" also includes a ring system in which the above heterocyclic group ring is fused to one or more carbocyclic groups, wherein the point of attachment is on the carbocyclic group or the heterocyclic group ring, or a ring system in which the above heterocyclic group ring is fused to one or more aryl or heteroaryl groups, wherein the point of attachment is on the heterocyclic group ring, and in such cases, the number of ring members continues to represent the number of ring members in the heterocyclic group ring system. Unless otherwise specified, 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- to 10-membered heterocyclic group. In some embodiments, the heterocyclic group is a substituted 3- to 10-membered heterocyclic group. In some embodiments, the heterocyclic group is a 5- to 10-membered non-aromatic ring system having ring carbon atoms and 1-4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, sulfur, boron, phosphorus, and silicon ("5- to 10-membered heterocyclic group"). In some embodiments, the heterocyclic group is a 5- to 8-membered non-aromatic ring system having ring carbon atoms and 1-4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur ("5- to 8-membered heterocyclic group"). In some embodiments, the heterocyclic group is a 5- to 6-membered non-aromatic ring system having ring carbon atoms and 1-4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur ("5- to 6-membered heterocyclic group"). In some embodiments, the 5- to 6-membered heterocyclic group has 1-3 ring heteroatoms selected from nitrogen, oxygen, and sulfur. In some embodiments, the 5- to 6-membered heterocyclic group has 1-2 ring heteroatoms selected from nitrogen, oxygen, and sulfur. In some embodiments, the 5- to 6-membered heterocyclic group has one ring heteroatom selected from nitrogen, oxygen, and sulfur.

[0050] "Hetero" when used to describe a compound or a group present in a compound 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 hydrocarbon groups: for example, alkyl, e.g., heteroalkyl; cycloalkyl, e.g., heterocyclic group; aryl, e.g., heteroaryl; cycloalkenyl, e.g., cycloheteroalkenyl, etc.; which have 1 to 5 heteroatoms, especially 1 to 3 heteroatoms.

[0051] "Alkoxy" means the group -OR, where R is a substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted carbocyclic group, substituted or unsubstituted heterocyclic group, substituted or unsubstituted aryl or substituted or unsubstituted heteroaryl. Specific alkoxies are methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, tert-butoxy, sec-butoxy, n-pentyloxy, n-hexyloxy and 1,2-dimethylbutoxy. Specific alkoxies are lower alkoxies, i.e., having 1 to 6 carbon atoms. Further specific alkoxies have 1-4 carbon atoms. Exemplary "substituted alkoxies" include, but are not limited to: -O-(CH 2 )t(C6-C10 aryl), -O-(CH2)t(5-10 membered heteroaryl), -O-(CH2)t(C3-C10 cycloalkyl) and -O-(CH2)t(4-10 membered heterocyclic group), where t is an integer from 0 to 4, and any aryl, heteroaryl, cycloalkyl or heterocyclic group present may itself be substituted by unsubstituted C1-C4 alkyl, halogen, unsubstituted C1-C4 alkoxy, unsubstituted C1-C4 haloalkyl, unsubstituted C1-C4 hydroxyalkyl, or unsubstituted C1-C4 haloalkoxy or hydroxy.

[0052] "Carboxy" means the group -C(O)OH.

[0053] "Cyano" means the group -CN.

[0054] "Halo" or "halogen" means fluorine (F), chlorine (Cl), bromine (Br) and iodine (I). In some embodiments, the halogen group is fluorine or chlorine.

[0055] "Hydroxy" means the group -OH.

[0056] "Nitro" means the group -NO2.

[0057] Other definitions

[0058] The term "pharmaceutically acceptable salts" refers to those salts that, within the scope of sound medical judgment, are suitable for contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response, etc., and are commensurate with a reasonable benefit / risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, pharmaceutically acceptable salts are described in detail by Berge et al. in J. Pharmaceutical Sciences (1977) 66:1-19. The pharmaceutically acceptable salts of the compounds of the present 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 the reaction of an amino group with an inorganic acid, such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, and perchloric acid, or with an organic acid, such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid, or malonic acid, or salts formed using methods employed in the art, such as, for example, an ion exchange method. Other pharmaceutically acceptable salts include: adipates, alginates, ascorbates, aspartates, benzenesulfonates, benzoates, bisulfates, borates, butyrates, camphorates, camphorsulfonates, citrates, cyclopentanepropionates, digluconates, dodecylsulfates, ethanesulfonates, formates, fumarates, gluconates, glycerophosphates, glucuronates, heptanoates, hexanoates, hydroiodides, 2-hydroxyethanesulfonates, lactates, lactobionates, laurates, laurylsulfates, malates, maleates, malonates, methanesulfonates, 2-naphthalenesulfonates, nicotinates, nitrates, oleates, oxalates, palmitates, pamoates, pectates, persulfates, 3-phenylpropionates, phosphates, picrates, pivalates, propionates, stearates, succinates, sulfates, tartrates, thiocyanates, p-toluenesulfonates, undecanoates, valerates, and the like. 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 salts, and the like. If appropriate, further pharmaceutically acceptable salts include non-toxic ammonium salts, quaternary ammonium salts, and amine cations formed using counterions, such as halide ions, hydroxide, carboxylate, sulfate, phosphate, nitrate, lower alkylsulfonate, and arylsulfonate. BRIEF DESCRIPTION OF THE DRAWINGS

[0059] Appendix Figure 1 It is a graph showing the changes in the plasma and brain contents of the reference substance over time.

[0060] Appendix Figure 2 It is a graph showing the changes in the plasma and brain contents of KH022 over time. DETAILED DESCRIPTION OF THE INVENTION

[0061] In order to better explain the technical solution of the present invention, the present invention provides synthetic or biological examples of some compounds, which do not further limit the scope of the invention.

[0062] Materials and Methods

[0063] Using the following general methods and procedures, the compounds provided herein can be prepared from readily available starting materials. It should be understood that under the given typical or preferred process conditions (i.e., reaction temperature, time, molar ratio of reactants, solvent, pressure, etc.), other process conditions can also be used unless otherwise specified. The optimal reaction conditions can vary depending on the specific reactants or solvents used, but such conditions can be determined by those skilled in the art through routine optimization.

[0064] In addition, it is obvious to those skilled in the art that conventional protecting groups may be required to prevent certain functional groups from undergoing unwanted reactions. The selection of suitable protecting groups for specific functional groups and the suitable conditions for protection and deprotection are well known in the art.

[0065] The compounds provided herein can be separated and purified by known standard methods. Such methods include (but are not limited to) recrystallization, column chromatography, HPLC, or supercritical fluid chromatography (SFC). Regarding the preparation details of the representative pyrazoles listed herein, the following reaction routes are provided. The compounds provided herein can be prepared by those skilled in the art of organic synthesis from known or commercially available starting materials and reagents.

[0066] The compounds of the present invention can be prepared according to the methods described in the art and using suitable reagents, starting materials, and purification methods known to those skilled in the art.

[0067] Synthesis of Compound KH001 in Example 1

[0068]

[0069] Compound KHC-2: Potassium cyanide (600.0 g, 9.2 mol) and acetic acid (640 mL) were added to a methanol solution (4 L) of compound KHC-1 (100.0 g, 0.3 mol), and the reaction mixture was stirred at room temperature overnight. Then the reaction solution was poured into water, extracted with ethyl acetate, the organic phase was dried over anhydrous sodium sulfate, and concentrated in vacuo. The resulting crude product was dissolved in acetic acid (2.5 L), and platinum dioxide (24.0 g, 0.1 mol) was added to the reaction solution, and hydrogenated with hydrogen. TLC showed that the reaction was complete. The reaction solution was filtered, the filtrate was concentrated to about 200 mL, diluted with 2.4 L of ice water, and an aqueous solution of sodium nitrite (46.0 g, 0.7 mol) was slowly added dropwise to the mixture. After the addition was completed, the mixture was stirred at room temperature for 15 hours. Then it was extracted with ethyl acetate, the combined organic layers were washed with brine, dried, and concentrated. The crude product was purified by silica gel column chromatography to obtain compound KHC-2 (57.3 g, yield 55%).

[0070] Compound KHC-3: Under a nitrogen atmosphere and at -78 °C, ethynylmagnesium bromide (395 mL, 0.5 M) was added dropwise to a solution of Compound KHC-2 (57.3 g, 164.5 mmol) in tetrahydrofuran (1 L). Then the temperature was gradually raised to room temperature and stirred overnight at room temperature. A saturated aqueous ammonium chloride solution was slowly added dropwise to the above mixture, followed by extraction with ethyl acetate. The organic phase was washed with brine, dried over anhydrous sodium sulfate, concentrated, and the resulting crude product was purified by silica gel column chromatography to obtain Compound KHC-3 (33.7 g, yield 55%).

[0071] Compound KHC-4: Compound KHC-3 (33.7 g, 90.5 mmol) was dissolved in a mixed solvent of acetic acid (200 mL) and water (20 mL), and then Dowex-50 (10.0 g) was added and the mixture was heated under reflux. After monitoring the completion of the reaction, it was cooled to room temperature, the resin was filtered, washed with ethyl acetate, the filtrate was neutralized with a 40% aqueous sodium hydroxide solution, the organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the resulting crude product was purified by silica gel column chromatography to obtain Compound KHC-4 (23.6 g, yield 70%).

[0072] Compound KHC-5: Compound KHC-4 (23.6 g, 63.3 mmol) and palladium black (1.2 g) (in tetrahydrofuran (200 mL)) were hydrogenated with hydrogen. After stirring overnight at room temperature, the mixture was filtered through a diatomaceous earth pad, washed with ethyl acetate, and the filtrate was concentrated in vacuo to give the crude compound. Recrystallization from acetone gave Compound KHC-5 (19.0 g, yield 80%).

[0073] Compound KH001: An aqueous sodium hydroxide solution (50 mL, 1 M) was slowly added dropwise to Compound KHC-5 (19.0 g, 50.8 mmol) in tetrahydrofuran (200 mL). After stirring overnight at room temperature, the pH was adjusted with dilute hydrochloric acid, extracted with ethyl acetate, the organic phase was washed with brine, dried over anhydrous sodium sulfate, concentrated, and the resulting crude product was purified by silica gel column chromatography to obtain Compound KHC (13.5 g, yield 80%). 1 H NMR (400 MHz, CDCl3), δ 4.06 (s, 1H), 2.32 - 2.28 (dd, 1H), 2.15 (s, 3H), 1.84 - 1.77 (m, 2H), 1.74 - 1.61 (m, 5H), 1.61 - 1.54 (m, 3H), 1.48 - 1.46 (m, 1H), 1.33 - 1.14 (m, 10H), 1.02 - 0.95 (m, 1H), 0.93 (s, 3H), 0.85 - 0.79 (m, 3H), 0.75 (s, 3H).

[0074] Example 2 Synthesis of Compound KH002

[0075]

[0076] Compound KHC-6: Dissolve the compound KH001 (1.5 g, 4.5 mmol) synthesized in Example 1 in methanol (40 mL) until clear. Stir and cool in an ice bath to an internal temperature of 5 °C, then slowly add bromine (1.5 g, 9.2 mmol). Remove the ice bath and react at room temperature for 4 hours. The orange color of the reaction solution turns light yellow. TLC monitoring shows that the reaction is complete. Add 30 mL of water to the reaction solution, extract with ethyl acetate (2 × 35 mL), combine the organic phases, wash with saturated brine, dry over anhydrous sodium sulfate, and evaporate the solvent under reduced pressure to obtain a crude light yellow solid. Purify by flash column chromatography on silica gel (eluted with dichloromethane) to obtain a white solid (1.0 g, 56.7%).

[0077] Compound KH002: Dissolve compound KHC-6 (80 mg, 0.2 mmol) in tetrahydrofuran (5 mL) until clear. Add K2CO3 (83 mg, 0.6 mmol) and 4-cyanopyrazole (56 mg, 0.6 mmol), and react overnight at room temperature. TLC monitoring shows that the reaction is complete. Add 10 mL of water to the reaction solution, extract with ethyl acetate (2 × 20 mL), combine the organic phases, wash with saturated brine, dry over anhydrous sodium sulfate, and evaporate the solvent under reduced pressure to obtain a crude light yellow solid. Purify by flash column chromatography on silica gel (eluted with dichloromethane / methanol = 100:1) and then recrystallize (ethyl acetate / petroleum ether = 1:10) to obtain a white solid (36 mg, 42.5%). 1 H NMR (400 MHz, CDCl3) δ 7.82 (s, 1H), 7.80 (s, 1H), 5.06 - 4.93 (m, 2H), 4.05 - 4.04 (m, 1H), 2.32 (dd, 1H), 1.88 - 1.83 (m, 2H), 1.79 - 1.66 (m, 4H), 1.64 - 1.58 (m, 2H), 1.54 - 1.44 (m, 4H), 1.39 - 1.13 (m, 9H), 1.04 - 0.97 (m, 1H), 0.94 (s, 3H), 0.85 - 0.75 (m, 3H), 0.74 (s, 3H).

[0078] Synthesis of Compound KH003 in Example 3

[0079]

[0080] Compound KH003: Compound KHC-6 (50 mg, 0.1 mmol) was dissolved in tetrahydrofuran (5 mL) until clear. Potassium carbonate (50 mg, 0.3 mmol) and 1H-triazole (25 mg, 0.3 mmol) were added. The reaction was carried out overnight at room temperature. TLC monitoring showed that the reaction was complete. 10 mL of water was added to the reaction solution, and it was extracted with ethyl acetate (2 × 20 mL). The combined organic phases were washed with saturated brine and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure to obtain a crude yellow solid. It was purified by flash chromatography on a silica gel column (eluted with dichloromethane / methanol = 150:1) and then recrystallized (ethyl acetate / petroleum ether = 1:10) to obtain a white solid (17 mg, 35.5%). 1 1H NMR (400 MHz, CDCl3) δ 7.76 (s, 1H), 7.60 (s, 1H), 5.31 - 5.17 (m, 2H), 4.06 - 4.05 (m, 1H), 2.37 (d, 1H), 1.87 - 1.61 (m, 8H), 1.52 - 1.10 (m, 13H), 1.04 - 0.98 (m, 1H), 0.95 (s, 3H), 0.92 - 0.79 (m, 3H), 0.74 (s, 3H).

[0081] Example 4 Synthesis of Compounds KH004 and KH005

[0082]

[0083] Compounds KH004 and KH005: Compound KHC-6 (103 mg, 0.2 mmol) was dissolved in acetonitrile (15 mL) until clear. Cesium carbonate (244 mg, 0.7 mmol) and 1H-tetrazole (53 mg, 0.7 mmol) were added. The reaction was carried out overnight at room temperature. TLC monitoring showed that the reaction was complete. 10 mL of water was added to the reaction solution, and it was extracted with ethyl acetate (2 × 25 mL). The combined organic phases were washed with saturated brine and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure to obtain a crude yellow solid. It was purified by flash chromatography on a silica gel column (eluted with dichloromethane / methanol = 150:1 to 90:1) to obtain two compounds, which were then recrystallized separately (ethyl acetate / petroleum ether = 1:10) to obtain white solid 1 (24 mg, 24.0%) and white solid 2 (36 mg, 36.0%). White solid 1 (KH004) 11H NMR (400 MHz, CDCl3) δ 8.57 (s, 1H), 5.50 (s, 2H), 4.06 - 4.05 (m, 1H), 2.35 (dd, 1H), 1.92 - 1.81 (m, 3H), 1.74 - 1.68 (m, 3H), 1.64 - 1.57 (m, 4H), 1.46 - 1.11 (m, 10H), 1.05 - 0.98 (m, 2H), 0.96 (s, 3H), 0.88 - 0.78 (m, 3H), 0.75 (s, 3H). White solid 2 (KH005) 1 1H NMR (400 MHz, CDCl3) δ 8.71 (s, 1H), 5.38 - 5.23 (m, 2H), 4.06 - 4.05 (m, 1H), 2.41 (dd, 1H), 2.28 (dd, 1H), 1.91 - 1.72 (m, 4H), 1.68 - 1.51 (m, 7H), 1.47 - 1.11 (m, 10H), 1.06 - 0.99 (m, 1H), 0.95 (s, 3H), 0.91 - 0.78 (m, 3H), 0.75 (s, 3H).

[0084] Synthesis of compound KH006 in Example 5

[0085]

[0086] Compound KH006: Compound KHC - 6 (50 mg, 0.1 mmol) was dissolved in acetonitrile (8 mL) until clear. Cs2CO3 (117 mg, 0.3 mmol) and 1H - pyrazole (24 mg, 0.3 mmol) were added. The reaction was carried out overnight at room temperature. TLC monitoring showed that the reaction was complete. 10 mL of water was added to the reaction solution, and it was extracted with ethyl acetate (2 × 20 mL). The combined organic phases were washed with saturated brine and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure to obtain a crude pale yellow solid. It was purified by flash chromatography on a silica gel column (eluted with dichloromethane / methanol = 100:1), and then recrystallized (ethyl acetate / petroleum ether = 1:10) to obtain a white solid (24 mg, 50.2%). 1 1H NMR (400 MHz, CDCl3) δ 7.56 (d, 1H), 7.39 (d, 1H), 6.34 (s, 1H), 5.03 - 4.92 (m, 2H), 4.05 - 4.04 (m, 1H), 2.28 (dd, 1H), 1.86 - 1.73 (m, 4H), 1.71 - 1.61 (m, 3H), 1.60 - 1.49 (m, 4H), 1.48 - 1.09 (m, 10H), 1.03 - 0.97 (m, 1H), 0.95 (s, 3H), 0.81 - 0.76 (m, 3H), 0.74 (s, 3H).

[0087] Synthesis of Compound KH007 in Example 6

[0088]

[0089] Compound KH007: Compound KHC-6 (150 mg, 0.3 mmol) was dissolved in tetrahydrofuran (10 mL) until clear, then K2CO3 (152 mg, 1.1 mmol) and 1H-imidazole (75 mg, 1.1 mmol) were added. The reaction was carried out overnight at room temperature. TLC monitoring showed that the reaction was complete. 10 mL of water was added to the reaction solution, and it was extracted with ethyl acetate (2×20 mL). The combined organic phases were washed with saturated brine and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure to obtain a crude yellow solid. It was purified by flash column chromatography on silica gel (eluted with dichloromethane / methanol = 30:1), and then recrystallized (with ethyl acetate) to obtain a white solid (35 mg, 24.1%). 1 H NMR (400 MHz, CDCl3) δ 7.47 (s, 1H), 7.11 (s, 1H), 6.85 (s, 1H), 4.74 (d, 2H), 4.06 - 4.05 (m, 1H), 2.30 (dd, 1H), 1.88 - 1.68 (m, 4H), 1.64 - 1.56 (m, 3H), 1.52 - 1.45 (m, 4H), 1.40 - 1.11 (m, 10H), 1.04 - 0.97 (m, 1H), 0.95 (s, 3H), 0.83 - 0.77 (m, 3H), 0.75 (s, 3H).

[0090] Synthesis of Compound KH008 in Example 7

[0091]

[0092] Compound KH008: Compound KHC-6 (100 mg, 0.2 mmol) was dissolved in tetrahydrofuran (10 mL) until clear, then K2CO3 (97 mg, 0.7 mmol) and 3-cyanopyrazole (68 mg, 0.7 mmol) were added. The reaction was carried out at 70 °C for 2 hours. TLC monitoring showed that the reaction was complete. 10 mL of water was added to the reaction solution, and it was extracted with ethyl acetate (2×20 mL). The combined organic phases were washed with saturated brine and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure to obtain a crude yellow solid. It was purified by preparative TLC chromatography (developing solvent: petroleum ether / acetone = 3:1), and then recrystallized (with acetonitrile) to obtain a white solid (20 mg, 19.4%). 11H NMR (400 MHz, CDCl3) δ 7.44 (d, 1H), 6.72 (d, 1H), 6.34 (s, 1H), 5.00 (q, 2H), 4.06 - 4.05 (m, 1H), 2.32 (dd, 1H), 1.88 - 1.68 (m, 4H), 1.63 - 1.60 (m, 3H), 1.50 - 1.45 (m, 4H), 1.41 - 1.11 (m, 10H), 1.04 - 0.97 (m, 1H), 0.95 (s, 3H), 0.87 - 0.79 (m, 3H), 0.75 (s, 3H).

[0093] Synthesis of Compound KH009 in Example 8

[0094]

[0095] Compound KH009: Compound KHC-6 (50 mg, 0.1 mmol) was dissolved in acetonitrile (5 mL) until clear. Cs2CO3 (120 mg, 0.3 mmol) and 1H-indole (68 mg, 0.7 mmol) were added. The reaction was carried out overnight at room temperature. TLC monitoring showed that the reaction was complete. 10 mL of water was added to the reaction solution, and it was extracted with ethyl acetate (2 × 20 mL). The combined organic phases were washed with saturated brine and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure to obtain a crude yellow solid. It was purified by preparative TLC chromatography (eluent: dichloromethane / ethyl acetate = 10:1) to obtain a light yellow solid (34 mg, 62.4%). 1 1H NMR (400 MHz, CDCl3) δ 7.65 - 7.63 (m, 1H), 7.21 - 7.18 (m, 1H), 7.13 - 7.10 (m, 2H), 7.01 - 7.00 (m, 1H), 6.57 (d, 1H), 4.83 (d, 2H), 4.06 - 4.05 (m, 1H), 2.30 (dd, 1H), 1.82 - 1.72 (m, 4H), 1.64 - 1.61 (m, 3H), 1.51 - 1.45 (m, 4H), 1.41 - 1.11 (m, 10H), 1.01 - 0.96 (m, 1H), 0.96 (s, 3H), 0.92 - 0.77 (m, 3H), 0.74 (s, 3H).

[0096] Synthesis of Compound KH010 in Example 9

[0097]

[0098] Compound KH010: Compound KHC-6 (100 mg, 0.2 mmol) was dissolved in acetonitrile (5 mL) until clear. K2CO3 (101 mg, 0.7 mmol) and 4-(trifluoromethyl)-1H-pyrazole (35 mg, 0.2 mmol) were added. The mixture was heated to 50 °C and reacted for 2 hours. TLC monitoring showed that the reaction was complete. 10 mL of water was added to the reaction solution, and it was extracted with ethyl acetate (2 × 20 mL). The combined organic phases were washed with saturated brine and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure to obtain a crude yellow solid. It was purified by flash chromatography on silica gel (eluent: petroleum ether / acetone = 7:1), and then recrystallized (acetonitrile) to obtain a white solid (61 mg, 53.1%). 1 H NMR (400 MHz, CDCl3) δ 7.71 (d, 2H), 4.98 (q, 2H), 4.06 - 4.05 (m, 1H), 2.32 (dd, 1H), 1.87 - 1.68 (m, 4H), 1.64 - 1.57 (m, 3H), 1.52 - 1.45 (m, 4H), 1.41 - 1.11 (m, 10H), 1.04 - 0.97 (m, 1H), 0.95 (s, 3H), 0.87 - 0.82 (m, 3H), 0.74 (s, 3H).

[0099] Synthesis of Compound KH011 in Example 10

[0100]

[0101] Compound KH011: Compound KHC-6 (50 mg, 0.1 mmol) was dissolved in acetonitrile (5 mL) until clear. Cs2CO3 (120 mg, 0.3 mmol) and 1H-indazole (43 mg, 0.3 mmol) were added. The mixture was stirred at room temperature overnight. TLC monitoring showed that the reaction was complete. 10 mL of water was added to the reaction solution, and it was extracted with ethyl acetate (2 × 20 mL). The combined organic phases were washed with saturated brine and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure to obtain a crude yellow solid. It was purified by preparative TLC chromatography (eluent: petroleum ether / acetone = 3:1), and then recrystallized (acetonitrile) to obtain a white solid (20 mg, 36.6%). 11H NMR (400 MHz, CDCl3) δ 8.11 (s, 1H), 7.78 (d, 1H), 7.45 - 7.41 (m, 1H), 7.23 - 7.19 (m, 2H), 5.31 - 5.20 (m, 2H), 4.06 - 4.05 (m, 1H), 2.35 (dd, 1H), 1.87–1.72 (m, 4H), 1.67 - 1.63 (m, 3H), 1.54 - 1.45 (m, 4H), 1.42 - 1.14 (m, 10H), 1.01 - 0.98 (m, 1H), 0.96 (s, 3H), 0.93 - 0.78 (m, 3H), 0.75 (s, 3H).

[0102] Synthesis of Compound KH012 in Example 11

[0103]

[0104] Compound KH012: Compound KHC - 6 (50 mg, 0.1 mmol) was dissolved in acetonitrile (5 mL) until clear. Cs2CO3 (120 mg, 0.3 mmol) and 1H - purine (44 mg, 0.3 mmol) were added. The mixture was stirred at room temperature overnight. TLC monitoring showed that the reaction was complete. 10 mL of water was added to the reaction solution, and it was extracted with ethyl acetate (2 × 20 mL). The combined organic phases were washed with saturated brine and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure to obtain a crude pale yellow solid. It was purified by preparative TLC chromatography (eluent: dichloromethane / methanol = 20:1), and then recrystallized (acetonitrile) to obtain a white solid (5 mg, 9.1%). 1 1H NMR (400 MHz, CDCl3) δ 9.22 (s, 1H), 9.06 (s, 1H), 8.25 (s, 1H), 5.18 (q, 2H), 4.07 - 4.06 (m, 1H), 2.49 (dd, 1H), 1.93 - 1.72 (m, 4H), 1.68 - 1.65 (m, 3H), 1.58 - 1.45 (m, 4H), 1.42 - 1.13 (m, 10H), 1.06 - 1.00 (m, 1H), 0.96 (s, 3H), 0.90 - 0.80 (m, 3H), 0.74 (s, 3H).

[0105] Synthesis of Compound KH013 in Example 12

[0106]

[0107] Compound KH013: Compound KHC-6 (80 mg, 0.2 mmol) was dissolved in acetonitrile (8 mL) until clear. Cs2CO3 (196 mg, 0.6 mmol) and 4-chloropyrazole (62 mg, 0.6 mmol) were added. The reaction was carried out overnight at room temperature. TLC monitoring showed that the reaction was complete. 15 mL of water was added to the reaction solution, and the mixture was extracted with ethyl acetate (2 * 30 mL). The combined organic phases were washed with saturated brine and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure to obtain a crude yellow solid. It was purified by flash chromatography on silica gel (eluted with dichloromethane / methanol = 100:1) and then recrystallized (ethyl acetate / petroleum ether = 1:10) to obtain a white solid (28 mg, 32.3%). 1 H NMR δ7.46(s,1H),7.38(s,1H),4.96 - 4.83(m,2H),4.06 - 4.05(m,1H),2.19(dd,1H),1.86 - 1.71(m,4H),1.64 - 1.55(m,3H),1.51 - 1.45(m,4H),1.45 - 1.10(m,10H),1.03 - 0.96(m,1H),0.95(s,3H),0.86 - 0.77(m,3H),0.74(s,3H).

[0108] Synthesis of Compound KH014 in Example 13

[0109]

[0110] Compound KH014: Compound KHC-6 (60 mg, 0.15 mmol) was dissolved in 5 mL of N,N-dimethylformamide until clear. 1M NaOH (1 mL, 1.00 mmol) was added. The reaction was carried out overnight at room temperature. TLC monitoring showed that the reaction was complete. 10 mL of water was added to the reaction solution, and the mixture was extracted with ethyl acetate (2 * 20 mL). The combined organic phases were washed with saturated sodium chloride solution and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure to obtain a crude product. It was purified using a thick preparative TLC silica gel plate (developed with dichloromethane / methanol = 50:1) to obtain a white solid (12 mg, 22.92%).

[0111] 1 H NMR(400MHz,DMSO-d6)δ4.94 - 4.89(m,1H),4.18(t,1H),4.06 - 4.02(m,2H),3.80(t,1H),2.27(dd,1H),1.77–0.65(m,27H),0.68(s,3H).

[0112] Synthesis of Compound KH015 in Example 14

[0113]

[0114] Compound KH015: Compound KHC-6 (60 mg, 0.15 mmol) was dissolved in 5 mL of acetonitrile until clear. Cs2CO3 (98 mg, 0.30 mmol) and 6-chloropurine (46 mg, 0.30 mmol) were added. The reaction was carried out overnight at room temperature. TLC monitoring showed that the reaction was complete. 10 mL of water was added to the reaction solution, and it was extracted with ethyl acetate (2×20 mL). The combined organic phases were washed with saturated sodium chloride solution and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure to obtain a crude product. The crude product was purified by preparative TLC silica gel thick plate (developed with dichloromethane / methanol = 40:1) to obtain a white solid (39 mg, 53.61%).

[0115] 1 H NMR (400 MHz, DMSO-d6) δ 8.60 (s, 1H), 8.37 (s, 1H), 5.41 (q, 2H), 4.20 (d, 1H), 3.79 (t, 1H), 2.56 (dd, 1H), 1.80–0.75 (m, 27H), 0.69 (s, 3H).

[0116] Synthesis of Compound KH016 in Example 15

[0117]

[0118] Compound KH016: Compound KHC-6 (60 mg, 0.15 mmol) was dissolved in 5 mL of acetonitrile until clear. Cs2CO3 (98 mg, 0.30 mmol) and 2-mercaptothiophene (17 mg, 0.30 mmol) were added. The reaction was carried out overnight at room temperature. TLC monitoring showed that the reaction was complete. 10 mL of water was added to the reaction solution, and it was extracted with ethyl acetate (2×20 mL). The combined organic phases were washed with saturated sodium chloride solution and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure to obtain a crude product. The crude product was purified by preparative TLC silica gel thick plate (developed with dichloromethane / methanol = 40:1) to obtain a white solid (19 mg, 28.34%).

[0119] 1 H NMR (400 MHz, CDCl3) δ 7.35 (d, 1H), 7.14 (d, 1H), 6.95 (dd, 1H), 4.04 (t, 1H), 3.62 (q, 2H), 2.45 (dd, 1H), 1.65–0.81 (m, 27H), 0.73 (s, 3H).

[0120] Synthesis of Compound KH017 in Example 16

[0121]

[0122] Compound KH017: Compound KHC-6 (60 mg, 0.15 mmol) was dissolved in 5 mL of acetonitrile until clear. Cs2CO3 (98 mg, 0.30 mmol) and 6-methoxypurine (45 mg, 0.30 mmol) were added. The reaction was carried out overnight at room temperature. TLC monitoring showed that the reaction was complete. 10 mL of water was added to the reaction solution, and it was extracted with ethyl acetate (2×20 mL). The combined organic phases were washed with saturated sodium chloride solution and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure to obtain the crude product. The crude product was purified by preparative TLC silica gel thick plate (developed with dichloromethane / methanol = 40:1) to obtain a white solid (46 mg, 63.76%).

[0123] 1 H NMR (400 MHz, DMSO-d6) δ 8.57 (s, 1H), 8.32 (s, 1H), 5.37 (q, 2H), 4.20 (d, 1H), 3.91 (s, 3H), 3.79 (t, 1H), 2.53 (dd, 1H), 1.79–0.75 (m, 27H), 0.69 (s, 3H).

[0124] Synthesis of Compounds KH018 and KH019 in Example 17

[0125]

[0126] Compounds KH018 and KH019: Compound KHC-6 (60 mg, 0.15 mmol) was dissolved in 5 mL of acetonitrile until clear. Cs2CO3 (98 mg, 0.30 mmol) and 5-methyl-1H-tetrazole (25 mg, 0.30 mmol) were added. The reaction was carried out overnight at room temperature. TLC monitoring showed that the reaction was complete. 10 mL of water was added to the reaction solution, and it was extracted with ethyl acetate (2×20 mL). The combined organic phases were washed with saturated sodium chloride solution and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure to obtain the crude product. The crude product was purified by preparative TLC silica gel thick plate (developed with dichloromethane / methanol = 40:1) to obtain white solid KH6070053-1 (13 mg, 20.88%) and white solid KH6070053-2 (27 mg, 43.37%) respectively.

[0127] 1 H NMR (400 MHz, CDCl3) δ 4.39 (s, 2H), 4.05 (t, 1H), 2.56 (s, 3H), 2.34 (dd, 1H), 1.92–

[0128] 0.76 (m, 27H), 0.74 (s, 3H).

[0129] 11H NMR (400 MHz, CDCl3) δ 5.15 (dd, 2H), 4.06 (t, 1H), 2.43 (s, 3H), 2.39 (dd, 1H), 1.95–

[0130] 0.78 (m, 27H), 0.75 (s, 3H).

[0131] Synthesis of Compound KH020 in Example 18

[0132]

[0133] Compound KH020: 60 mg (0.15 mmol) of compound KHC-6 was added to 5 mL of acetonitrile and dissolved to clarity. Cs2CO3 (98 mg, 0.30 mmol) and 2-oxa-6-aza-spiro[3.3]heptane (30 mg, 0.30 mmol) were added. The reaction was carried out overnight at room temperature. TLC monitoring showed that the reaction was complete. 10 mL of water was added to the reaction solution, and the mixture was extracted with ethyl acetate (2 × 20 mL). The combined organic phases were washed with saturated sodium chloride solution and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure to obtain a crude product. The crude product was purified by preparative TLC silica gel thick plate (developed with dichloromethane / methanol = 40:1) to obtain a white solid (21 mg, 32.56%).

[0134] 1 1H NMR (400 MHz, CDCl3) δ 4.75 (s, 4H), 4.04 (t, 1H), 3.57 (dd, 4H), 3.40 (dd, 2H), 2.13 (dd, 1H), 1.90–0.75 (m, 27H), 0.73 (s, 3H).

[0135] Synthesis of Compound KH021 in Example 19

[0136]

[0137] Compound KHC-7: 0.70 g (2.11 mmol) of compound KH001 was added to 100 mL of dichloromethane and dissolved. DMP (1.78 g, 4.20 mmol) was added in batches. The reaction was carried out overnight at room temperature under N2 protection. TLC monitoring showed that the reaction was complete. 40 mL of water was added to the reaction solution, and the mixture was extracted with ethyl acetate (2 × 80 mL). The combined organic phases were washed with saturated sodium chloride solution and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure to obtain a crude product. The crude product was purified by flash column chromatography on silica gel (eluted with petroleum ether / acetone = 40:1 → 30:1) to obtain white solid KHC-7 (0.62 g, 89.47%).

[0138] Compound KH021: Take FeCl3 (0.61 g, 3.75 mmol) and LiCl (0.34 g, 8.07 mmol) into a 100 mL three-necked flask, under N2 protection, add 40 mL of anhydrous tetrahydrofuran, stir at room temperature for 5 minutes, then transfer to -45 °C, slowly dropwise add CH3MgBr (15.20 mL, 15.20 mmol), stir at about -40 °C for 15 minutes; dropwise add an anhydrous tetrahydrofuran solution of compound KHC-7 (0.62 g, 1.88 mmol), after the addition is complete, warm up to about -20 °C and stir the reaction for 2 hours. TLC monitoring shows that the reaction is complete. Add about 40 mL of saturated NH4Cl to quench the reaction solution, extract with ethyl acetate (2×80 mL), combine the organic phases, wash with saturated sodium chloride solution, dry over anhydrous sodium sulfate, and evaporate the solvent under reduced pressure to obtain the crude product. Purify by flash chromatography on a silica gel column (eluting with petroleum ether / acetone = 40:1 → 35:1) to obtain white solid KH021 (0.52 g, 78.50%).

[0139] 1 H NMR (400 MHz, CDCl3) 2.29 (dd, 1H), 2.13 (s, 3H), 1.88–1.25 (m, 21H), 1.20 (s, 3H), 0.95 (s, 3H), 0.88–0.78 (m, 3H), 0.70 (s, 3H).

[0140] Example 20 Synthesis of Compounds KH022 and KH023

[0141]

[0142]

[0143] Compounds KHC-8 and KHC-9: Take compound KH021 (1.25 g, 3.60 mmol), dissolve it in 40 mL of methanol until clear, and then dropwise add bromine (0.86 g, 5.40 mmol). React at room temperature for 4 hours, the orange color of the reaction solution turns light yellow, and TLC monitoring shows that the reaction is complete. Add 30 mL of water to the reaction solution, extract with ethyl acetate (2×65 mL), combine the organic phases, wash with saturated sodium chloride solution, dry over anhydrous sodium sulfate, and evaporate the solvent under reduced pressure to obtain the crude product of light yellow solid. Purify by flash chromatography on a silica gel column (eluting with dichloromethane) to obtain white solid KHC-8 (0.93 g, 60.78%) and white solid KHC-9 (0.36 g, 19.89%).

[0144] Compounds KH022 and KH023: Compound KHC-8 (100 mg, 0.24 mmol) was dissolved in 5 mL of acetonitrile until clear. Cs2CO3 (234 mg, 0.72 mmol) and 1H-tetrazole (50 mg, 0.72 mmol) were added. The reaction was carried out overnight at room temperature. TLC monitoring showed that the reaction was complete. 10 mL of water was added to the reaction solution, and it was extracted with ethyl acetate (2×20 mL). The combined organic phases were washed with saturated sodium chloride solution and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure to obtain the crude product. It was purified using a thick preparative TLC silica gel plate (developed with dichloromethane / methanol = 30:1) to obtain white solid KH022 (20 mg, 20.08%) and white solid KH023 (47 mg, 47.19%) respectively.

[0145] 1 1H NMR (400 MHz, CDCl3) δ 8.56 (s, 1H), 5.54–5.45 (m, 2H), 2.35 (dd, 1H), 1.89–1.25 (m, 21H), 1.20 (s, 3H), 0.95 (s, 3H), 0.88–0.80 (m, 3H), 0.72 (s, 3H).

[0146] 1 1H NMR (400 MHz, CDCl3) δ 8.71 (s, 1H), 5.30 (dd, 2H), 2.35 (dd, 1H), 1.83–1.23 (m, 21H), 1.20 (s, 3H), 0.95 (s, 3H), 0.87–0.78 (m, 3H), 0.72 (s, 3H).

[0147] Example 21 Synthesis of Compound KH024

[0148]

[0149] Compound KH024: Compound KHC-8 (60 mg, 0.14 mmol) was dissolved in 5 mL of acetonitrile until clear. Cs2CO3 (91 mg, 0.28 mmol) and 3-cyanopyrazole (26 mg, 0.28 mmol) were added. The reaction was carried out overnight at room temperature. TLC monitoring showed that the reaction was complete. 10 mL of water was added to the reaction solution, and it was extracted with ethyl acetate (2×20 mL). The combined organic phases were washed with saturated sodium chloride solution and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure to obtain the crude product. It was purified using a thick preparative TLC silica gel plate (developed with dichloromethane / methanol = 40:1) to obtain a white solid (16 mg, 26.09%).

[0150] 11H NMR (400 MHz, CDCl3) δ 7.45 (d, 1H), 6.73 (d, 1H), 5.06 (q, 2H), 2.30 (dd, 1H), 1.86–

[0151] 1.23 (m, 21H), 1.20 (s, 3H), 0.95 (s, 3H), 0.87–0.77 (m, 3H), 0.72 (s, 3H).

[0152] Synthesis of Compound KH025 in Example 22

[0153]

[0154] Compound KH025: Take compound KHC-8 (100 mg, 0.24 mmol), add 5 mL of dimethyl sulfoxide to dissolve and clarify, add Cs2CO3 (235 mg, 0.72 mmol) and 3-hydroxybenzisoxazole (97 mg, 0.72 mmol). React at room temperature overnight. TLC monitoring shows that the reaction is complete. Add 10 mL of water to the reaction solution, extract with ethyl acetate (2 × 20 mL), combine the organic phases, wash with saturated sodium chloride solution, dry over anhydrous sodium sulfate, and evaporate the solvent under reduced pressure to obtain the crude product. Purify with a thick preparative TLC silica gel plate (developed with petroleum ether / acetone = 5:1) to obtain a white solid (70 mg, 60.76%).

[0155] 1 1H NMR (400 MHz, CDCl3) δ 7.72 (d, 1H), 7.58–7.49 (m, 1H), 7.42 (d, 1H), 7.29 (d, 1H), 5.11–4.95 (m, 2H), 2.35 (dd, 1H), 1.87–1.22 (m, 21H), 1.20 (s, 3H), 0.99 (s, 3H), 0.87–0.80 (m, 3H), 0.72 (s, 3H).

[0156] Synthesis of Compound KH026 in Example 23

[0157]

[0158] Compound KH026: Take compound KHC-8 (53 mg, 0.12 mmol), dissolve it in 5 mL of acetonitrile until clear, and add K2CO3 (100 mg, 0.72 mmol) and 2-mercaptobenzoxazole (24 mg, 0.16 mmol). React overnight at room temperature. TLC monitoring shows that the reaction is complete. Add 10 mL of water to the reaction solution, extract with ethyl acetate (2 * 20 mL), combine the organic phases, wash with saturated sodium chloride solution, dry over anhydrous sodium sulfate, and evaporate the solvent under reduced pressure to obtain the crude product. Purify it using a thick preparative TLC silica gel plate (developed with petroleum ether / acetone = 6:1) to obtain a white solid (30 mg, 50.40%).

[0159] 1 H NMR (400 MHz, CDCl3) δ 7.55 (d, 1H), 7.42 (d, 1H), 7.34 - 7.24 (m, 2H), 4.33 (m, 2H), 2.54 (dd, 1H), 1.83–1.23 (m, 21H), 1.18 (s, 3H), 0.94 (s, 3H), 0.85–0.78 (m, 3H), 0.70 (s, 3H).

[0160] Synthesis of Compound KH027 in Example 24

[0161]

[0162] Compound KH027: Take compound KHC-9 (110 mg, 0.22 mmol), dissolve it in 10 mL of acetonitrile until clear, and add Cs2CO3 (215 mg, 0.66 mmol) and 4-cyanopyrazole (61 mg, 0.66 mmol). React overnight at room temperature. TLC monitoring shows that the reaction is complete. Add 10 mL of water to the reaction solution, extract with ethyl acetate (2 * 20 mL), combine the organic phases, wash with saturated sodium chloride solution, dry over anhydrous sodium sulfate, and evaporate the solvent under reduced pressure to obtain the crude product. Purify it using a thick preparative TLC silica gel plate (developed with dichloromethane / methanol = 70:1) to obtain a white solid (76 mg, 65.30%).

[0163] 1 H NMR (400 MHz, CDCl3) δ 8.23 (s, 1H), 8.13 (s, 1H), 7.92 (s, 1H), 7.83 (s, 1H), 7.03 (s, 1H), 2.11 (dd, 1H), 1.83–1.23 (m, 21H), 1.20 (s, 3H), 0.91 (s, 3H), 0.82–0.75 (m, 3H), 0.70 (s, 3H).

[0164] Synthesis of Compound KH028 in Example 25

[0165]

[0166] Compound KH028: Take compound KHC-8 (70 mg, 0.16 mmol), add 8 mL of dimethyl sulfoxide to dissolve and clarify, then add Cs2CO3 (160 mg, 0.49 mmol) and 2-hydroxybenzoxazole (66 mg, 0.49 mmol). React at room temperature overnight. TLC monitoring shows that the reaction is complete. Add 10 mL of water to the reaction solution, extract with ethyl acetate (2 * 20 mL), combine the organic phases, wash with saturated sodium chloride solution, dry over anhydrous sodium sulfate, and evaporate the solvent under reduced pressure to obtain the crude product. Purify using a thick preparative TLC silica gel plate (developed with dichloromethane / methanol = 60:1) to obtain a white solid (55 mg, 71.61%).

[0167] 1 H NMR (400 MHz, CDCl3) δ 7.24–7.18 (m, 1H), 7.17–7.06 (m, 2H), 6.75–6.64 (m, 1H), 4.58 (dd, 2H), 2.38 (dd, 1H), 1.92–1.22 (m, 21H), 1.20 (s, 3H), 0.97 (s, 3H), 0.89 - 0.78 (m, 3H), 0.72 (s, 3H).

[0168] Synthesis of Compound KH029 in Example 26

[0169]

[0170] Compound KH029: Take compound KHC-8 (60 mg, 0.14 mmol), add 5 mL of dimethyl sulfoxide to dissolve and clarify, then add Cs2CO3 (138 mg, 0.42 mmol) and 8-methyl-4-hydroxyquinazoline (68 mg, 0.42 mmol). React at room temperature overnight. TLC monitoring shows that the reaction is complete. Add 10 mL of water to the reaction solution, extract with ethyl acetate (2 * 20 mL), combine the organic phases, wash with saturated sodium chloride solution, dry over anhydrous sodium sulfate, and evaporate the solvent under reduced pressure to obtain the crude product. Purify using a thick preparative TLC silica gel plate (developed with dichloromethane / methanol = 60:1) to obtain a white solid (50 mg, 70.72%).

[0171] 11H NMR (400 MHz, CDCl3) δ 8.13 (d, 1H), 7.90 (s, 1H), 7.61 (d, 1H), 7.39 (t, 1H), 4.81 (s, 2H), 2.63 (s, 3H), 2.43 (dd, 1H), 1.88–1.22 (m, 21H), 1.20 (s, 3H), 0.97 (s, 3H), 0.90–0.81 (m, 3H), 0.72 (s, 3H).

[0172] Synthesis of Compound KH030 in Example 27

[0173]

[0174] Compound KH030: Take compound KHC-8 (50 mg, 0.12 mmol), add 5 mL of dimethyl sulfoxide to dissolve and clarify, add Cs2CO3 (78 mg, 0.24 mmol) and 3-hydroxy-4-methyl-5-methoxy-4H-1,2,4-triazole (31 mg, 0.24 mmol). React at room temperature overnight. TLC monitoring shows that the reaction is complete. Add 10 mL of water to the reaction solution, extract with ethyl acetate (2×20 mL), combine the organic phases, wash with saturated sodium chloride solution, dry over anhydrous sodium sulfate, and evaporate the solvent under reduced pressure to obtain the crude product. Purify with a thick preparative TLC silica gel plate (developed with petroleum ether / acetone = 4:1) to obtain a white solid (20 mg, 35.16%).

[0175] 1 1H NMR (400 MHz, CDCl3) δ 4.47 (q, 2H), 3.93 (s, 3H), 3.12 (s, 3H), 2.28 (dd, 1H), 1.86–

[0176] 1.22 (m, 21H), 1.19 (s, 3H), 0.94 (s, 3H), 0.85–0.78 (m, 3H), 0.70 (s, 3H).

[0177] Synthesis of Compound KH031 in Example 28

[0178]

[0179] Compound KH031: Take compound KHC-8 (60 mg, 0.14 mmol), add 5 mL of acetonitrile to dissolve and clarify, then add K2CO3 (58 mg, 0.42 mmol) and 1-methyl-5-mercapto-1H-tetrazole (68 mg, 0.42 mmol). React at room temperature overnight. TLC monitoring shows that the reaction is complete. Add 10 mL of water to the reaction solution, extract with ethyl acetate (2 * 20 mL), combine the organic phases, wash with saturated sodium chloride solution, dry over anhydrous sodium sulfate, and evaporate the solvent under reduced pressure to obtain the crude product. Purify using a thick preparative TLC silica gel plate (developed with petroleum ether / acetone = 4:1) to obtain a white solid (50 mg, 77.47%).

[0180] 1 H NMR (400 MHz, CDCl3) 4.40 (dd, 2H), 3.97 (s, 3H), 2.47 (dd, 1H), 1.88–1.22 (m, 21H), 1.19 (s, 3H), 0.93 (s, 3H), 0.87–0.79 (m, 3H), 0.71 (s, 3H).

[0181] Example 29 Synthesis of Compound KH032

[0182]

[0183] Compound KH032: Take compound KHC-8 (50 mg, 0.12 mmol), add 5 mL of dimethyl sulfoxide to dissolve and clarify, then add K2CO3 (50 mg, 0.36 mmol) and 2-hydroxy-1H-imidazole (30 mg, 0.36 mmol). React at room temperature overnight. TLC monitoring shows that the reaction is complete. Add 10 mL of water to the reaction solution, extract with ethyl acetate (2 * 20 mL), combine the organic phases, wash with saturated sodium chloride solution, dry over anhydrous sodium sulfate, and evaporate the solvent under reduced pressure to obtain the crude product. Purify using a thick preparative TLC silica gel plate (developed with petroleum ether / acetone = 2:1) to obtain a white solid (11 mg, 21.37%).

[0184] 1 H NMR (400 MHz, CDCl3) δ 9.26 (s, 1H), 6.32 (s, 1H), 6.17 (s, 1H), 4.53–4.32 (m, 2H), 2.31 (dd, 1H), 1.86–1.25 (m, 21H), 1.21 (s, 3H), 0.95 (s, 3H), 0.88–0.78 (m, 3H), 0.71 (s, 3H).

[0185] Example 30 Synthesis of Compound KH033

[0186]

[0187] Compound KH033: Take compound KHC-8 (60 mg, 0.14 mmol), add 5 mL of dimethyl sulfoxide to dissolve and clarify, then add Cs2CO3 (138 mg, 0.42 mmol) and 2-hydroxyquinoline (61 mg, 0.42 mmol). React at room temperature overnight. TLC monitoring shows that the reaction is complete. Add 10 mL of water to the reaction solution, extract with ethyl acetate (2 × 20 mL), combine the organic phases, wash with saturated sodium chloride solution, dry over anhydrous sodium sulfate, and evaporate the solvent under reduced pressure to obtain the crude product. Purify using a thick preparative TLC silica gel plate (developed with petroleum ether / acetone = 4:1) to obtain a white solid (15 mg, 21.87%).

[0188] 1 H NMR (400 MHz, CDCl3) δ 8.05 (d, 1H), 7.75 (dd, 2H), 7.61 (t, 1H), 7.39 (t, 1H), 7.05 (d, 1H), 5.15 (s, 2H), 2.43 (dd, 1H), 1.98–1.23 (m, 21H), 1.21 (s, 3H), 0.97 (s, 3H), 0.91–0.78 (m, 3H), 0.74 (s, 3H).

[0189] Synthesis of Compound KH034 in Example 31

[0190]

[0191] Compound KH034: Take compound KHC-8 (60 mg, 0.14 mmol), add 5 mL of dimethyl sulfoxide to dissolve and clarify, then add K2CO3 (58 mg, 0.42 mmol) and 3-hydroxy-1H-1,2,4-triazole (36 mg, 0.42 mmol). React at room temperature overnight. TLC monitoring shows that the reaction is complete. Add 10 mL of water to the reaction solution, extract with ethyl acetate (2 × 20 mL), combine the organic phases, wash with saturated sodium chloride solution, dry over anhydrous sodium sulfate, and evaporate the solvent under reduced pressure to obtain the crude product. Purify using a thick preparative TLC silica gel plate (developed with dichloromethane / methanol = 40:1) to obtain a white solid (40 mg, 66.45%).

[0192] 1 H NMR (400 MHz, CDCl3) δ 9.30 (s, 1H), 7.42 (s, 1H), 4.47 (q, 2H), 2.33 (dd, 1H), 1.87–

[0193] 1.25 (m, 21H), 1.20 (s, 3H), 0.95 (s, 3H), 0.87–0.79 (m, 3H), 0.71 (s, 3H).

[0194] Synthesis of Compound KH035 in Example 32

[0195]

[0196] Compound KH035: Take compound KHC-8 (80 mg, 0.19 mmol), add 6 mL of dimethyl sulfoxide to dissolve and clarify, then add Cs2CO3 (184 mg, 0.56 mmol) and 8-fluoro-4-hydroxyquinazoline (92 mg, 0.56 mmol). React at room temperature overnight. TLC monitoring shows that the reaction is complete. Add 10 mL of water to the reaction solution, extract with ethyl acetate (2 × 20 mL), combine the organic phases, wash with saturated sodium chloride solution, dry over anhydrous sodium sulfate, and evaporate the solvent under reduced pressure to obtain the crude product. Purify with a thick preparative TLC silica gel plate (developed with petroleum ether / acetone = 2:1) to obtain a white solid (65 mg, 67.21%).

[0197] 1 H NMR (400 MHz, CDCl3) δ 8.06 (d, 1H), 7.88 (s, 1H), 7.55–7.40 (m, 2H), 4.79 (m, 2H), 2.43 (dd, 1H), 1.88–1.22 (m, 21H), 1.21 (s, 3H), 0.96 (s, 3H), 0.90–0.81 (m, 3H), 0.72 (s, 3H).

[0198] Synthesis of Compound KH036 in Example 33

[0199]

[0200] Compound KH036: Take compound KHC-8 (60 mg, 0.14 mmol), add 5 mL of dimethyl sulfoxide to dissolve and clarify, then add K2CO3 (58 mg, 0.42 mmol) and 6-fluoropyrido[3,4-d]pyrimidin-4-one (69 mg, 0.42 mmol). React at room temperature overnight. TLC monitoring shows that the reaction is complete. Add 10 mL of water to the reaction solution, extract with ethyl acetate (2 × 20 mL), combine the organic phases, wash with saturated sodium chloride solution, dry over anhydrous sodium sulfate, and evaporate the solvent under reduced pressure to obtain the crude product. Purify with a thick preparative TLC silica gel plate (developed with dichloromethane / methanol = 40:1) to obtain a white solid (40 mg, 56.02%).

[0201] 11H NMR (400 MHz, CDCl3) δ 8.82 (s, 1H), 7.80 (s, 1H), 7.66 (d, 1H), 4.80 (q, 2H), 2.43 (dd, 1H), 1.89–1.23 (m, 21H), 1.20 (s, 3H), 0.95 (s, 3H), 0.90–0.80 (m, 3H), 0.71 (s, 3H).

[0202] Synthesis of Compound KH037 in Example 34

[0203]

[0204] Compound KH037: Take compound KHC-8 (60 mg, 0.14 mmol), add 5 mL of acetonitrile to dissolve and clarify, add K2CO3 (58 mg, 0.42 mmol) and 4-methyl-4H-3-mercapto-1,2,4-triazole (49 mg, 0.42 mmol). React at room temperature overnight. TLC monitoring shows that the reaction is complete. Add 10 mL of water to the reaction solution, extract with ethyl acetate (2×20 mL), combine the organic phases, wash with saturated sodium chloride solution, dry over anhydrous sodium sulfate, and evaporate the solvent under reduced pressure to obtain the crude product. Purify with a thick preparative TLC silica gel plate (developed with petroleum ether / acetone = 3:2) to obtain a white solid (53 mg, 82.30%).

[0205] 1 1H NMR (400 MHz, CDCl3) δ 8.16 (s, 1H), 4.34 (dd, 2H), 3.65 (s, 3H), 2.45 (dd, 1H), 1.88–1.22 (m, 21H), 1.19 (s, 3H), 0.91 (s, 3H), 0.87–0.79 (m, 3H), 0.70 (s, 3H).

[0206] Synthesis of Compound KH038 in Example 35

[0207]

[0208] Compound KH038: Take compound KHC-8 (60 mg, 0.14 mmol), add 5 mL of dimethyl sulfoxide to dissolve and clarify, add Cs2CO3 (138 mg, 0.42 mmol) and 3-hydroxyisoxazole (36 mg, 0.42 mmol). React at room temperature overnight. TLC monitoring shows that the reaction is complete. Add 10 mL of water to the reaction solution, extract with ethyl acetate (2×20 mL), combine the organic phases, wash with saturated sodium chloride solution, dry over anhydrous sodium sulfate, and evaporate the solvent under reduced pressure to obtain the crude product. Purify by flash chromatography on a silica gel column (eluted with petroleum ether / acetone = 15:1) to obtain a white solid (39 mg, 65.00%).

[0209] 1 1H NMR (300 MHz, CDCl3) δ 8.12 (d, 1H), 6.06 (d, 1H), 4.87 (s, 2H), 2.29 (dd, 1H), 1.90–1.22 (m, 21H), 1.20 (s, 3H), 0.97 (s, 3H), 0.90–0.74 (m, 3H), 0.71 (s, 3H).

[0210] Synthesis of Compound KH039 in Example 36

[0211]

[0212] Compound KH039: Take compound KHC-8 (75 mg, 0.18 mmol), add 5 mL of acetonitrile to dissolve and clarify, then add Cs2CO3 (171 mg, 0.53 mmol) and 5-phenyltetrazole (77 mg, 0.53 mmol). React at room temperature overnight. TLC monitoring shows that the reaction is complete. Add 10 mL of water to the reaction solution, extract with ethyl acetate (2 × 20 mL), combine the organic phases, wash with saturated sodium chloride solution, dry over anhydrous sodium sulfate, and evaporate the solvent under reduced pressure to obtain the crude product. Purify with a thick preparative TLC silica gel plate (developed with petroleum ether / acetone = 3:1) to obtain a white solid (79 mg, 89.39%).

[0213] 1 1H NMR (400 MHz, CDCl3) δ 8.19–8.07 (m, 2H), 7.55–7.42 (m, 3H), 5.54–5.40 (m, 2H), 2.37 (dd, 1H), 1.92–1.23 (m, 21H), 1.21 (s, 3H), 0.97 (s, 3H), 0.90–0.78 (m, 3H), 0.73 (s, 3H).

[0214] Synthesis of Compound KH040 in Example 37

[0215]

[0216] Compound KH040: Take compound KHC-8 (60 mg, 0.14 mmol), add 5 mL of acetonitrile to dissolve and clarify, then add Cs2CO3 (91 mg, 0.28 mmol) and 4-cyanopyrazole (26 mg, 0.28 mmol). React at room temperature overnight. TLC monitoring shows that the reaction is complete. Add 10 mL of water to the reaction solution, extract with ethyl acetate (2 × 20 mL), combine the organic phases, wash with saturated sodium chloride solution, dry over anhydrous sodium sulfate, and evaporate the solvent under reduced pressure to obtain the crude product. Purify with a thick preparative TLC silica gel plate (developed with dichloromethane / methanol = 40:1) to obtain a white solid (16 mg, 26.09%).

[0217] 1 1H NMR (400 MHz, CDCl3) δ 7.82 (s, 1H), 7.81 (s, 1H), 5.00 (q, 2H), 2.32 (dd, 1H), 1.88–

[0218] 1.22 (m, 21H), 1.20 (s, 3H), 0.94 (s, 3H), 0.87–0.79 (m, 3H), 0.71 (s, 3H).

[0219] Synthesis of Compound KH041 in Example 38

[0220]

[0221] Compound KH041: Take compound KHC-8 (70 mg, 0.16 mmol), dissolve it in 5 mL of acetonitrile until clear, add Cs2CO3 (160 mg, 0.49 mmol) and 2-mercaptothiophene (57 mg, 0.49 mmol). React at room temperature overnight. TLC monitoring shows that the reaction is complete. Add 10 mL of water to the reaction solution, extract with ethyl acetate (2 × 20 mL), combine the organic phases, wash with saturated sodium chloride solution, dry over anhydrous sodium sulfate, and evaporate the solvent under reduced pressure to obtain the crude product. Purify by flash chromatography on a silica gel column (eluted with dichloromethane / petroleum ether = 1:1) to obtain an off-white solid (27 mg, 35.53%).

[0222] 1 1H NMR (400 MHz, CDCl3) δ 7.35 (dd, 1H), 7.14 (dd, 1H), 6.95 (dd, 1H), 3.62 (dd, 2H), 1.90–1.21 (m, 21H), 1.20 (s, 3H), 0.90 (s, 3H), 0.84 - 0.74 (m, 3H), 0.70 (s, 3H).

[0223] Synthesis of Compound KH042 in Example 39

[0224]

[0225] Compound KH042: Take compound KHC-8 (150 mg, 0.35 mmol), add 30 mL of acetone, 15 mL of water and K2CO3 (49 mg, 0.35 mmol), and dissolve until clear. React under reflux at 70 °C overnight. TLC monitoring shows that the reaction is complete. Add 10 mL of water to the reaction solution, extract with ethyl acetate (2 * 30 mL), combine the organic phases, wash with saturated sodium chloride solution, dry over anhydrous sodium sulfate, and evaporate the solvent under reduced pressure to obtain the crude product. Purify using a thick preparative TLC silica gel plate (developed with dichloromethane / methanol = 50:1) to obtain a white solid (21 mg, 16.54%).

[0226] 1 H NMR (400 MHz, DMSO-d6) δ 4.92 (t, 1H), 4.06 - 4.04 (m, 2H), 3.85 (t, 1H), 2.27 (dd, 1H), 1.82–1.10 (m, 21H), 1.05 (s, 3H), 0.87 (s, 3H), 0.85 - 0.70 (m, 3H), 0.65 (s, 3H).

[0227] Synthesis of Compounds KH043 and KH044 in Example 39

[0228]

[0229] Compounds KH043 and KH044: Take compound KHC-8 (60 mg, 0.14 mmol), dissolve it in 5 mL of acetonitrile until clear, add Cs2CO3 (91 mg, 0.28 mmol) and 1H-triazole (15 mg, 0.21 mmol). React at room temperature overnight. TLC monitoring shows that the reaction is complete. Add 10 mL of water to the reaction solution, extract with ethyl acetate (2 * 20 mL), combine the organic phases, wash with saturated sodium chloride solution, dry over anhydrous sodium sulfate, and evaporate the solvent under reduced pressure to obtain the crude product. Purify using a thick preparative TLC silica gel plate (developed with dichloromethane / methanol = 40:1) to obtain white solid KH6070100-1 (12 mg, 20.70%) and white solid KH6070100-2 (27 mg, 46.58%) respectively.

[0230] 1 H NMR (400 MHz, CDCl3) δ 7.68 (s, 2H), 5.34–5.20 (m, 2H), 2.25 (dd, 1H), 1.88–1.23 (m, 21H), 1.20 (s, 3H), 0.95 (s, 3H), 0.89–0.79 (m, 3H), 0.71 (s, 3H).

[0231] 11H NMR (400 MHz, CDCl3) δ 7.76 (s, 1H), 7.61 (s, 1H), 5.25 (dd, 2H), 2.37 (dd, 1H), 1.86–1.22 (m, 21H), 1.20 (s, 3H), 0.94 (s, 3H), 0.86–0.76 (m, 3H), 0.71 (s, 3H).

[0232] Synthesis of Compound KH045 in Example 40

[0233] Compound KHC-11: Take compound KHC-10 (150.0 g, 183.8 mmol), dissolve it in 500 mL of tetrahydrofuran, add 10% Pd / C (6.0 g), replace the reaction solution with H2 three times and carry out the hydrogenation reaction, stir at room temperature for 24 hours. Filter the reaction solution, evaporate the solvent under reduced pressure from the filtrate to obtain the crude product. Pulp with a petroleum ether:acetone = 1:1 solution to obtain compound KHC-11 (150.0 g, 99.26%).

[0234] Compound KHC-12: Take compound KHC-11 (150.0 g, 547.4 mmol), dissolve it in 1000 mL of methanol, add I2 (14.0 g, 54.8 mmol), heat and stir at 60 °C for 12 hours, concentrate the reaction solution, and purify it by column chromatography (PE:EA = 10:1) to obtain white solid compound KHC-12 (130.0 g, 74.20%).

[0235] Compound KHC-13: Take compound KHC-12 (130.0 g, 406.2 mmol), dissolve it in 1000 mL of N,N-dimethylformamide, add t-BuOK (91.0 g, 812.4 mmol) and trimethylsulfonium iodide (165.8 g, 812.7 mmol), stir the reaction solution at room temperature, and react overnight under N2 protection. Add brine to the reaction solution, extract with ethyl acetate (1.0 L × 3), combine the organic phases, dry, and remove the solvent to obtain white solid compound KHC-13 (120.0 g, 88.50%).

[0236] Compound KHC-14: Take compound KHC-13 (120.0 g, 359.4 mmol), dissolve it in 800 mL of ethanol and 160 mL of water, add sodium azide (70.2 mg, 1077.9 mmol) and ammonium chloride (67.2 g, 1257.9 mmol), heat and stir overnight at 90 °C. Add brine to the reaction solution, extract with ethyl acetate (1.0 L × 3), combine the organic phases, dry, remove the solvent, and purify by column chromatography (PE:EA = 10:1) to obtain white solid compound KHC-14 (130.0 g, 95.59%).

[0237] Compound KHC-15: Take compound KHC-14 (125.6 g, 332.4 mmol), dissolve it in 1000 mL of acetonitrile, add sodium iodide (249.3 g, 1662.0 mmol), stir at room temperature for 0.5 hour, dropwise add TMSCl (144.9 g, 1329.6 mmol), and react the reaction solution at room temperature for 4 hours. Add brine to the reaction solution, extract with ethyl acetate (1000 mL × 3), combine the organic phases, dry, remove the solvent, and purify by column chromatography (PE:EA = 10:1) to obtain white solid compound KHC-15 (72.0 g, 75.20%). Compound KHC-16: Take compound KHC-15 (72.0 g, 250.0 mmol), dissolve it in 1000 mL of methanol, add I2 (19.0 g, 75.0 mmol), heat and stir at 60 °C for 12 hours, concentrate the reaction solution, and purify by column chromatography (PE:EA = 10:1) to obtain white solid compound KHC-16 (53.0 g, 63.50%).

[0238] Compound KHC-17: Take trimethylsilylacetylene (77.8 g, 793.0 mmol), add 1000 mL of tetrahydrofuran, add n-butyllithium solution (1.6 M, 329.0 mmol, 205.6 ml) at -78 °C, and stir for 2 hours. Take compound KHC-16 (53.0 g, 158.6 mmol) and dissolve it in 400 mL of tetrahydrofuran, add it to the reaction solution, and stir at -78 °C for 2 hours. Gradually warm the reaction solution to 0 °C and pour it into saturated ammonium chloride solution, extract with ethyl acetate (800 mL × 3). Concentrate the reaction solution, and purify by column chromatography (PE:EA = 10:1) to obtain white solid compound KHC-17 (56.0 g, 81.55%).

[0239] Compound KHC-18: Take compound KHC-17 (56.0 g, 129.3 mmol), dissolve it in 800 mL of tetrahydrofuran, dropwise add 1.0 M TBAF solution (129.3 mL, 129.3 mmol) at 0 °C, and stir and react for 1 hour. Add brine to the reaction solution, extract with ethyl acetate (1000 mL × 3), combine the organic phases, dry, remove the solvent, and purify by column chromatography (PE:EA = 5:1) to obtain white solid compound KHC-18 (43.0 g, 92.30%).

[0240] Compound KHC-19: Dissolve compound KHC-18 (38.0 g, 105.6 mmol) in 800 mL of tetrahydrofuran, add 1 N HCl solution to adjust the acidity to pH = 3, and stir at room temperature for 12 hours. Add brine (1.0 L) to the reaction solution, extract with ethyl acetate (1000 mL × 3), combine the organic phases, dry, remove the solvent, and purify by column chromatography (PE:EA = 5:1) to obtain white solid compound KHC-19 (32.0 g, 96.50%).

[0241] Compound KHC-20: Take methanesulfonic acid (148.0 g, 1541.6 mmol) into 1000 mL of dichloromethane. Take compound KHC-19 (32.0 g, 101.9 mmol) dissolved in 1000 mL of dichloromethane and drop it into the reaction solution. Heat to 60 °C and stir for 2 hours. Cool the reaction solution to room temperature, add brine (1.0 L), extract with dichloromethane (1000 mL × 3), combine the organic phases, dry, remove the solvent, and purify by column chromatography (PE:EA = 10:1) to obtain white solid compound KHC-20 (20.0 g, 62.50%). Compound KHC-21: Take compound KHC-20 (20.0 g, 63.7 mmol), dissolve it in 500 mL of tetrahydrofuran, add 10% Pd / C (3.0 g), replace the reaction solution with H2 three times for hydrogenation reaction, and stir at room temperature for 2 days. Filter the reaction solution, evaporate the solvent under reduced pressure from the filtrate to obtain the crude product. Purify by column chromatography (PE:EA = 15:1) to obtain white solid compound KHC-21 (17.0 g, 84.40%).

[0242] Compound KHC045: Take FeCl3 (23.0 g, 142.0 mmol) and LiCl (13.5 g, 321.4 mmol) into a 2000 mL three-necked flask, protect with N2, add 800 mL of anhydrous tetrahydrofuran, stir at room temperature for 20 minutes, then transfer to -40 °C, and slowly dropwise add CH3MgBr (3 M, 190 mL, 570.0 mmol). Stir at about -40 °C for 30 minutes; dropwise add a solution of compound KHC-21 (17.0 g, 53.8 mmol) in anhydrous tetrahydrofuran (200 mL). After the addition is complete, warm up to about -20 °C and stir for 4 hours. TLC monitoring shows that the reaction is complete. Quench the reaction solution with about 40 mL of saturated NH4Cl, extract with ethyl acetate (1.0 L × 3), wash the combined organic phases with saturated sodium chloride solution, dry with anhydrous sodium sulfate, evaporate the solvent under reduced pressure to obtain the crude product. Purify by column chromatography (PE:EA = 15:1) to obtain white solid compound KH045 (10.2 g, 57.10%).

[0243] 11H NMR (400 MHz, CDCl3) δ 2.34–2.25 (m, 1H), 2.14 (s, 3H), 2.00 (t, J = 11.4 Hz, 1H), 1.87–1.78 (m, 1H), 1.75–1.54 (m, 9H), 1.51–1.43 (m, 2H), 1.39–1.30 (m, 4H), 1.27–1.13 (m, 7H), 1.09–0.84 (m, 8H).

[0244]

[0245] Synthesis of Compound KH046 in Example 41

[0246]

[0247] Compounds KHC-22 and KHC-23: Take compound KH045 (1.50 g, 4.50 mmol), add 40 mL of methanol to dissolve and clarify, then dropwise add bromine (1.08 g, 6.75 mmol). React at room temperature for 4 hours. The color of the reaction solution changes from orange to light yellow. TLC monitoring shows that the reaction is complete. Add 30 mL of water to the reaction solution, extract with ethyl acetate (2 × 65 mL), combine the organic phases, wash with saturated sodium chloride solution, dry over anhydrous sodium sulfate, and evaporate the solvent under reduced pressure to obtain a crude yellow solid. Purify by flash column chromatography on silica gel (eluted with dichloromethane) to obtain white solid KHC-22 (1.19 g, 64.34%) and white solid KHC-23 (0.33 g, 17.84%).

[0248] Compound KHC046: Take compound KHC-22 (50 mg, 0.12 mmol), add 5 mL of dimethyl sulfoxide to dissolve and clarify, add K2CO3 (21 mg, 0.15 mmol) and 8-fluoro-4-hydroxyquinazoline (25 mg, 0.15 mmol). React at room temperature overnight. TLC monitoring shows that the reaction is complete. Add 10 mL of water to the reaction solution, extract with ethyl acetate (2 × 20 mL), combine the organic phases, wash with saturated sodium chloride solution, dry over anhydrous sodium sulfate, and evaporate the solvent under reduced pressure to obtain a crude product. Purify by preparative TLC silica gel thick plate (developed with petroleum ether / acetone = 4:1) to obtain a white solid (33 mg, 55.56%).

[0249] 1 1H NMR (400 MHz, CDCl3) δ 8.06 (d, 1H), 7.88 (s, 1H), 7.55–7.40 (m, 2H), 4.80 (q, 2H), 2.44 (dd, 1H), 2.04 - 2.01 (m, 1H), 1.91 - 1.88 (m, 1H), 1.84–1.16 (m, 21H), 1.15–0.80 (m, 8H).

[0250] Synthesis of Compound KH047 in Example 42

[0251]

[0252] Compound KH047: Take compound KHC-22 (50 mg, 0.12 mmol), add 5 mL of acetonitrile to dissolve and clarify, add Cs2CO3 (49 mg, 0.15 mmol) and 5-mercapto-1-methyl-1H-tetrazole (17 mg, 0.15 mmol). React overnight at room temperature. TLC monitoring shows that the reaction is complete. Add 10 mL of water to the reaction solution, extract with ethyl acetate (2 × 20 mL), combine the organic phases, wash with saturated sodium chloride solution, dry over anhydrous sodium sulfate, and evaporate the solvent under reduced pressure to obtain the crude product. Purify with a thick preparative TLC silica gel plate (developed with petroleum ether / acetone = 4:1) to obtain a white solid (51 mg, 50.12%).

[0253] 1 H NMR (400 MHz, CDCl3) 4.40 (dd, 2H), 3.97 (s, 3H), 2.48 (dd, 1H), 2.05 - 1.99 (m, 1H), 1.93 - 1.82 (m, 1H), 1.83–1.14 (m, 21H), 1.13–0.70 (m, 8H).

[0254] Synthesis of Compound KH048 in Example 43

[0255]

[0256] Compound KH048: Take compound KHC-22 (60 mg, 0.14 mmol), add 5 mL of dimethyl sulfoxide to dissolve and clarify, add K2CO3 (58 mg, 0.42 mmol) and 6-fluoropyrido[3,4-d]pyrimidin-4-one (69 mg, 0.42 mmol). React overnight at room temperature. TLC monitoring shows that the reaction is complete. Add 10 mL of water to the reaction solution, extract with ethyl acetate (2 × 20 mL), combine the organic phases, wash with saturated sodium chloride solution, dry over anhydrous sodium sulfate, and evaporate the solvent under reduced pressure to obtain the crude product. Purify with a thick preparative TLC silica gel plate (developed with dichloromethane / methanol = 40:1) to obtain a white solid (37 mg, 53.28%).

[0257] 11H NMR (400 MHz, CDCl3) δ 8.82 (s, 1H), 7.79 (s, 1H), 7.66 (d, 1H), 4.80 (dd, 2H), 2.44 (dd, 1H), 2.05 - 1.99 (m, 1H), 1.93 - 1.82 (m, 1H), 1.83–1.14 (m, 21H), 1.13–0.70 (m, 8H).

[0258] Synthesis of Compound KH049 in Example 44

[0259]

[0260] Compound KH049: 60 mg (0.15 mmol) of compound KHC-22 was dissolved in 5 mL of acetonitrile until clear. Cs2CO3 (62 mg, 0.19 mmol) and 2-mercaptobenzoxazole (29 mg, 0.19 mmol) were added. The reaction was carried out overnight at room temperature. TLC monitoring showed that the reaction was complete. 10 mL of water was added to the reaction solution, and it was extracted with ethyl acetate (2 × 20 mL). The combined organic phases were washed with saturated sodium chloride solution and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure to obtain a crude product. It was purified by preparative TLC silica gel thick plate (developed with petroleum ether / acetone = 4:1) to obtain a white solid (42 mg, 42.92%).

[0261] 1 1H NMR (400 MHz, CDCl3) δ 7.55 (d, 1H), 7.42 (d, 1H), 7.34 - 7.24 (m, 2H), 4.33 (dd, 2H), 2.57 (dd, 1H), 2.05 - 1.99 (m, 1H), 1.93 - 1.82 (m, 1H), 1.83–1.14 (m, 21H), 1.13–0.70 (m, 8H).

[0262] Synthesis of Compound KH050 in Example 45

[0263]

[0264] Compound KH050: Compound KHC-22 (60 mg, 0.15 mmol) was dissolved in 5 mL of acetonitrile until clear. Cs2CO3 (62 mg, 0.19 mmol) and 3-mercapto-4-methyl-4H-1,2,4-triazole (22 mg, 0.19 mmol) were added. The reaction was carried out overnight at room temperature. TLC monitoring showed that the reaction was complete. 10 mL of water was added to the reaction solution, and the mixture was extracted with ethyl acetate (2×20 mL). The combined organic phases were washed with saturated sodium chloride solution and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure to obtain a crude product. The crude product was purified by preparative TLC silica gel thick plate (developed with dichloromethane / methanol = 70:1) to obtain a white solid (17 mg, 25.41%).

[0265] 1 H NMR (400 MHz, CDCl3) δ 8.22 (s, 1H), 4.37 (dd, 2H), 3.68 (s, 3H), 2.50 (dd, 1H), 2.10–1.97 (m, 1H), 2.06-2.02 (m, 1H), 1.79–1.14 (m, 21H), 1.12–0.79 (m, 8H).

[0266] Example 46 Synthesis of Compound KH051

[0267]

[0268] Compound KH051: Take compound KHC-22 (60 mg, 0.14 mmol), dissolve it in 5 mL of dimethyl sulfoxide until clear, add K2CO3 (58 mg, 0.42 mmol) and 3-hydroxy-1H-1,2,4-triazole (36 mg, 0.42 mmol). The reaction was carried out overnight at room temperature. TLC monitoring showed that the reaction was complete. 10 mL of water was added to the reaction solution, and the mixture was extracted with ethyl acetate (2×20 mL). The combined organic phases were washed with saturated sodium chloride solution and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure to obtain a crude product. The crude product was purified by preparative TLC silica gel thick plate (developed with dichloromethane / methanol = 40:1) to obtain a white solid (27 mg, 46.36%).

[0269] 1 H NMR (400 MHz, CDCl3) δ 9.57 (s, 1H), 7.42 (s, 1H), 4.46 (q, 2H), 2.34 (dd, 1H), 2.07–

[0270] 1.96 (m, 1H), 1.88-1.82 (m, 1H), 1.79 (s, 21H), 1.11–0.81 (m, 8H).

[0271] Example 47 Synthesis of Compounds KH052 and KH053

[0272]

[0273] Compounds KH052 and KH053: Compound KHC-22 (100 mg, 0.24 mmol) was dissolved in 5 mL of acetonitrile until clear. Cs2CO3 (234 mg, 0.72 mmol) and 1H-tetrazole (50 mg, 0.72 mmol) were added. The reaction was carried out overnight at room temperature. TLC monitoring showed that the reaction was complete. 10 mL of water was added to the reaction solution, and the mixture was extracted with ethyl acetate (2×20 mL). The combined organic phases were washed with saturated sodium chloride solution and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure to obtain the crude product. The crude product was purified by preparative TLC silica gel thick plate (developed with dichloromethane / methanol = 30:1), and white solids KH6070069-1 (19 mg, 19.74%) and KH6070069-2 (44 mg, 45.72%) were obtained respectively.

[0274] 1 H NMR (400 MHz, CDCl3) δ 8.57 (s, 1H), 5.49 (s, 2H), 2.33 (dd, 1H), 2.05 - 2.01 (m, 1H), 1.94–1.84 (m, 1H), 1.83–1.15 (m, 21H), 1.14–0.82 (m, 8H).

[0275] 1 H NMR (400 MHz, CDCl3) δ 8.72 (s, 1H), 5.29 (dd, 2H), 2.41 (dd, 1H), 2.06 - 2.01 (m, 1H), 1.95–1.85 (m, 1H), 1.85–1.15 (m, 21H), 1.14–0.82 (m, 8H).

[0276] Synthesis of Compound KH054 in Example 48

[0277]

[0278] Compound KH054: Compound KHC-22 (60 mg, 0.15 mmol) was dissolved in 5 mL of acetonitrile until clear. Cs2CO3 (62 mg, 0.19 mmol) and 2-hydroxyoxazole[4,5-b]pyridine (26 mg, 0.19 mmol) were added. The reaction was carried out overnight at room temperature. TLC monitoring showed that the reaction was complete. 10 mL of water was added to the reaction solution, and the mixture was extracted with ethyl acetate (2×20 mL). The combined organic phases were washed with saturated sodium chloride solution and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure to obtain the crude product. The crude product was purified by preparative TLC silica gel thick plate (developed with dichloromethane / methanol = 70:1), and a white solid (17 mg, 42.92%) was obtained.

[0279] 1 1H NMR (400 MHz, CDCl3) δ 7.25 (d, 1H), 7.01 (d, 1H), 6.69 (t, 1H), 5.11 (q, 2H), 2.40 (dd, 1H), 2.08–1.98 (m, 1H), 1.93–1.83 (m, 1H), 1.82–1.14 (m, 21H), 1.15–0.81 (m, 8H).

[0280] Synthesis of Compound KH055 in Example 49

[0281]

[0282] Compound KH055: To a solution of compound KHC-22 (100 mg, 0.24 mmol) in 5 mL of acetonitrile was added Cs2CO3 (156 mg, 0.48 mmol) and 4-cyanopyrazole (45 mg, 0.48 mmol). The reaction was stirred overnight at room temperature. TLC monitoring showed that the reaction was complete. The reaction mixture was added to 10 mL of water and extracted with ethyl acetate (2 × 20 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 to obtain a crude product. The crude product was purified by preparative TLC silica gel thick plate (developed with dichloromethane / methanol = 30:1) to obtain a white solid (51 mg, 50.12%).

[0283] 1 1H NMR (400 MHz, CDCl3) δ 7.83 (s, 1H), 7.81 (s, 1H), 4.99 (q, 2H), 2.32 (dd, 1H), 2.05-2.01 (m, 1H), 1.92–1.83 (m, 1H), 1.82–1.15 (m, 21H), 1.14–0.81 (m, 8H).

[0284] Synthesis of Compound KH056 in Example 50

[0285]

[0286] Compound KH056: To a solution of compound KHC-23 (40 mg, 0.10 mmol) in 5 mL of acetonitrile was added Cs2CO3 (98 mg, 0.30 mmol) and 4-cyanopyrazole (28 mg, 0.30 mmol). The reaction was stirred overnight at room temperature. TLC monitoring showed that the reaction was complete. The reaction mixture was added to 10 mL of water and extracted with ethyl acetate (2 × 20 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 to obtain a crude product. The crude product was purified by preparative TLC silica gel thick plate (developed with dichloromethane / methanol = 30:1) to obtain a white solid (12 mg, 23.30%).

[0287] 1 1H NMR (400 MHz, CDCl3) δ 8.22 (s, 1H), 8.14 (s, 1H), 7.92 (s, 1H), 7.84 (s, 1H), 7.02 (s, 1H), 2.10 (dd, 1H), 2.05 - 2.00 (m, 1H), 1.87–1.78 (m, 1H), 1.78–1.14 (m, 21H), 1.12–0.79 (m, 8H).

[0288] Synthesis of Compound KH057 in Example 51

[0289]

[0290] Compound KH057: 60 mg (0.15 mmol) of compound KHC-22 was added to 5 mL of dimethyl sulfoxide and dissolved to clarity. Cs2CO3 (143 mg, 0.44 mmol) and 2-hydroxybenzoxazole (59 mg, 0.44 mmol) were added. The reaction was carried out overnight at room temperature. TLC monitoring showed that the reaction was complete. 10 mL of water was added to the reaction solution, and it was extracted with ethyl acetate (2 × 20 mL). The combined organic phases were washed with saturated sodium chloride solution and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure to obtain a crude product. It was purified by preparative TLC silica gel thick plate (developed with petroleum ether / acetone = 3:1) to obtain a white solid (30 mg, 42.92%).

[0291] 1 1H NMR (400 MHz, CDCl3) δ 7.24–7.19 (m, 1H), 7.17–7.07 (m, 2H), 6.75–6.65 (m, 1H), 4.58 (s, 2H), 2.40 (dd, 1H), 2.08–1.98 (m, 1H), 1.93–1.83 (m, 1H), 1.82–1.14 (m, 21H), 1.15–0.81 (m, 8H).

[0292] Synthesis of Compound KH058 in Example 52

[0293]

[0294] Compound KH058: Take compound KHC-22 (80 mg, 0.19 mmol), add 5 mL of dimethyl sulfoxide to dissolve and clarify, then add Cs2CO3 (190 mg, 0.58 mmol) and 8-methyl-4-quinazolinone (93 mg, 0.58 mmol). React at room temperature overnight. TLC monitoring shows that the reaction is complete. Add 10 mL of water to the reaction solution, extract with ethyl acetate (2×20 mL), combine the organic phases, wash with saturated sodium chloride solution, dry over anhydrous sodium sulfate, and evaporate the solvent under reduced pressure to obtain the crude product. Purify using a thick preparative TLC silica gel plate (developed with petroleum ether / acetone = 3:1) to obtain a white solid (28 mg, 30.01%).

[0295] 1 H NMR (400 MHz, CDCl3) δ 8.13 (d, 1H), 7.88 (s, 1H), 7.61 (d, 1H), 7.39 (t, 1H), 4.80 (dd, 2H), 3.49 (s, 3H), 2.45 (dd, 1H), 2.03–2.01 (m, 1H), 1.92–1.84 (m, 1H), 1.83–1.16 (m, 21H), 1.14–0.80 (m, 8H).

[0296] Synthesis of Compound KH059 in Example 53

[0297]

[0298] Compound KH059: Take compound KHC-22 (80 mg, 0.19 mmol), add 5 mL of dimethyl sulfoxide to dissolve and clarify, then add Cs2CO3 (190 mg, 0.58 mmol) and 8-hydroxy-1,7-naphthyridine (85 mg, 0.58 mmol). React at room temperature overnight. TLC monitoring shows that the reaction is complete. Add 10 mL of water to the reaction solution, extract with ethyl acetate (2×20 mL), combine the organic phases, wash with saturated sodium chloride solution, dry over anhydrous sodium sulfate, and evaporate the solvent under reduced pressure to obtain the crude product. Purify using a thick preparative TLC silica gel plate (developed with petroleum ether / acetone = 3:1) to obtain a white solid (28 mg, 30.89%).

[0299] 1 H NMR (400 MHz, CDCl3) δ 8.86 (d, 1H), 7.88 (d, 1H), 7.54 (dd, 1H), 6.96 (d, 1H), 6.45 (d, 1H), 4.86 (s, 2H), 2.45 (dd, 1H), 2.09–1.95 (m, 1H), 1.87 - 1.83 (m, 1H), 1.80–1.15 (m, 21H), 1.12–0.76 (m, 8H).

[0300] Synthesis of Compound KH060 in Example 54

[0301]

[0302] Compound KH060: Take compound KHC-22 (100 mg, 0.24 mmol), add 5 mL of dimethyl sulfoxide to dissolve and clarify, then add Cs2CO3 (156 mg, 0.48 mmol) and 3-hydroxy-4-methyl-5-methoxy-4H-1,2,4-triazole (62 mg, 0.48 mmol). React at room temperature overnight. TLC monitoring shows that the reaction is complete. Add 10 mL of water to the reaction solution, extract with ethyl acetate (2 × 20 mL), combine the organic phases, wash with saturated sodium chloride solution, dry over anhydrous sodium sulfate, and evaporate the solvent under reduced pressure to obtain the crude product. Purify using a thick preparative TLC silica gel plate (developed with petroleum ether / acetone = 4:1) to obtain a white solid (56 mg, 50.72%).

[0303] 1 H NMR (400 MHz, CDCl3) δ 4.54–4.40 (m, 2H), 3.93 (s, 3H), 3.12 (s, 3H), 2.29 (dd, 1H), 2.03-1.99 (m, 1H), 1.85-1.82 (m, 1H), 1.77–1.13 (m, 21H), 1.10–0.81 (m, 8H).

[0304] Synthesis of Compound KH061 in Example 55

[0305]

[0306] Compound KH061: Take compound KHC-22 (80 mg, 0.19 mmol), add 5 mL of dimethyl sulfoxide to dissolve and clarify, then add Cs2CO3 (190 mg, 0.58 mmol) and 2-hydroxyquinoline (84 mg, 0.58 mmol). React at room temperature overnight. TLC monitoring shows that the reaction is complete. Add 10 mL of water to the reaction solution, extract with ethyl acetate (2 × 20 mL), combine the organic phases, wash with saturated sodium chloride solution, dry over anhydrous sodium sulfate, and evaporate the solvent under reduced pressure to obtain the crude product. Purify using a thick preparative TLC silica gel plate (developed with petroleum ether / acetone = 4:1) to obtain a white solid (45 mg, 49.76%).

[0307] 11H NMR (400 MHz, CDCl3) δ 8.05 (d, 1H), 7.75 (dd, 2H), 7.61 (t, 1H), 7.39 (t, 1H), 7.05 (d, 1H), 5.15 (s, 2H), 2.43 (dd, 1H), 2.04 - 2.00 (m, 1H), 1.86 - 1.83 (m, 1H), 1.78–1.12 (m, 21H), 1.10–0.80 (m, 8H).

[0308] Example 56 In vitro cell activity assay

[0309] The structures of the comparative compounds used in the present invention are as follows:

[0310]

[0311] The specific synthesis process was prepared with reference to CN105339381B.

[0312] The present invention uses recombinant GABA A patch-clamp electrophysiology methods of two subtypes, α1β2γ2 and α4β3δ, of the receptor to evaluate the in vitro activity of the compounds.

[0313] 56.1 Electrophysiological detection solution

[0314] Composition of extracellular and intracellular solutions:

[0315] Extracellular solution: 140 mM NaCl, 5 mM CsCl, 2 mM CaCl2·2H2O, 1 mM MgCl2·6H2O, 5 mM HEPES, 10 mM D-Glucose, pH = 7.4 (NaOH).

[0316] Intracellular solution: 130 mM CsCl, 0.1 mM CaCl2·2H2O, 2 mM MgCl2·6H2O, 1.1 mM EGTA, 5 mM Na 2- ATP, 10 mM HEPES, pH = 7.2 (CsOH).

[0317] After the prepared intracellular solution was dispensed into 1 mL tubes each, it was stored frozen at -20 °C in a refrigerator. Freshly thawed electrode internal solution was used for each experiment, and all intracellular solutions were used up within three months.

[0318] 56.2 Cell lines with stable ion channel expression:

[0319]

[0320] 56.3 Cell culture

[0321] 56.3.1 GABA A(α1β2γ2) Cell Culture

[0322] Stable expression of GABA A The HEK293 cell line stably expressing the (α1β2γ2) receptor was cultured in DMEM medium containing 10% fetal bovine serum, 800 μg / mL G418, 200 μg / mL Hygromycin B, and 100 μg / mL Zeocin at a culture temperature of 37°C and a carbon dioxide concentration of 5%.

[0323] Cell passage: Remove the old medium and wash once with PBS, then add 1 mL of 0.25%-Trypsin-EDTA solution and incubate at 37°C for 0.5 min. When the cells detach from the bottom of the dish, add 5 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 1000 rpm for 5 min to collect the cells. For amplification or maintenance culture, inoculate the cells into a 6-cm cell culture dish, and the cell seeding amount for each cell culture dish is 2.5×10 5 cells (final volume: 5 mL).

[0324] To maintain the electrophysiological activity of the cells, the cell density must not exceed 80%.

[0325] Before the experiment, the cells were separated with 0.25%-Trypsin-EDTA, and 8×10 3 cells were seeded onto cover slips and cultured in a 24-well plate (final volume: 500 μL). After 18 h, patch clamp detection was performed on the cells.

[0326] 56.3.2 GABA A (α4β3δ) Cell Culture

[0327] HEK293 cells were cultured in DMEM medium containing 10% fetal bovine serum at a culture temperature of 37°C and a carbon dioxide concentration of 5%.

[0328] Cell passage: Remove the old medium and wash once with PBS, then add 1 mL of 0.25%-Trypsin-EDTA solution and incubate at 37°C for 0.5 min. When the cells detach from the bottom of the dish, add 5 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 1000 rpm for 5 min to collect the cells. For amplification or maintenance culture, inoculate the cells into a 6-cm cell culture dish, and the cell seeding amount for each dish is 2.5×10 5 (final volume: 5 mL). Cells for transfection and patch clamp detection were inoculated into a 24-well plate with pre-placed cover slips, 8×10 3 cells per well.

[0329] Transfection: Transfect GABA on the second day A4 (The ratio of the three plasmids of α4, β3, and δ is 1:1:1). Use the X-tremeGENE HP DAN Transfection Reagent. The ratio of plasmid to transfection reagent is 1 μg: 2 μL. The dosage of plasmid per well in a 24-well plate is 0.5 μg, and the dosage of transfection reagent is 1 μL. The following are the specific steps for transfecting 12 wells: Add 600 μL of Opti-MEM to a sterile centrifuge tube, add 6 μg of plasmid, and mix well; then add 12 μL of transfection reagent, mix well, and incubate at room temperature for 15 min. Then add the transfection complex dropwise to the cells, 50 μL per well, and gently shake to mix well.

[0330] Change the medium on the third day and perform patch clamp detection on the fourth day.

[0331] 56.4 Patch clamp detection

[0332] Use a microelectrode puller (P97, Sutter Instruments) to pull the capillary glass tube (BF150-86-10, Sutter Instruments) into a recording electrode. Under an inverted microscope (IX71, Olympus), operate the microelectrode manipulator (MP285, Sutter Instruments) to bring the recording electrode into contact with the cell, and apply negative pressure suction to form a GΩ seal. After forming a GΩ seal, perform rapid capacitance compensation, and then continue to apply negative pressure to break the cell membrane to form a whole-cell recording mode. Then perform slow capacitance compensation and record the membrane capacitance and series resistance. Do not apply leakage compensation.

[0333] Place the cover glass with cells in the recording bath in the inverted microscope. The test article working solution and the external solution without the compound are perfused from low concentration to high concentration in sequence through the recording bath by gravity to act on the cells, and liquid exchange is performed using a vacuum pump during recording. Multiple data are detected and repeated for each concentration. All electrophysiological experiments are carried out at room temperature.

[0334] Record GABA using whole-cell patch clamp A1 The voltage stimulation protocol for receptor current is as follows: When the whole-cell seal is formed, the cell membrane voltage is clamped at -70 mV. Record GABA current in Gap-free mode A1 When the cell is stable, start to stimulate with 3 μΜ GABA. After incubating for 30 s at the detected concentration, sequentially add a mixture of 3 μΜ GABA and different concentrations of the compound, and finally add 100 μΜ GABA. The time interval between each drug administration is 2 min. The experimental data are collected by an EPC-10 amplifier (HEKA) and stored in the PatchMaster (HEKA) software.

[0335] Whole-cell patch clamp recording of GABA A4 The voltage stimulation protocol for receptor current is as follows: When a whole-cell seal is formed, the cell membrane voltage is clamped at -70 mV. Record the GABA A4 current in Gap-free mode. After the cell is stable, start to stimulate with 10 nM GABA. After incubating for 30 s at the detected concentration, sequentially administer a mixture of 10 nM GABA and different concentrations of the compound, and finally administer 10 μΜ GABA. The time interval between each drug administration is 2 min. The experimental data is collected by an EPC-10 amplifier (HEKA) and stored in the PatchMaster (HEKA) software.

[0336] 56.5 Data quality criteria

[0337] The following criteria are used to determine whether the data is acceptable:

[0338] (1) Electrode resistance < 5 MΩ;

[0339] (2) Seal resistance > 1 GΩ;

[0340] (3) Access resistance at the beginning < 15 MΩ;

[0341] (4) Access resistance at the end < 15 MΩ;

[0342] (5) The current has no obvious spontaneous decay;

[0343] (6) There is no obvious leakage current at a membrane potential of -70 mV;

[0344] 56.6 Data analysis

[0345] When detecting GABA A1 the currents of 3 μΜ GABA and the mixtures with different drug concentrations are normalized to the current of 100 μΜ GABA Then calculate the activation rate corresponding to each drug concentration.

[0346] When detecting GABA A4 the currents of 10 nM GABA and the mixtures with different drug concentrations are normalized to the current of 10 μM GABA Then calculate the activation rate corresponding to each drug concentration. Calculate the mean and standard error for each concentration. And calculate the half-activation concentration of each compound using the following equation:

[0347] Y = Bottom + Bottom / (1 + 10^((LogEC 50 -X) × HillSlope))

[0348] The dose-dependent effect was non-linearly fitted with the above equation, where EC 50 is the half-activation concentration and Hillslope represents the Hill coefficient. Curve fitting and the calculation of EC 50 were completed using Graphpad 5.0 software.

[0349] 56.7 In vitro activity results

[0350] Table 1: Electrophysiological evaluation of the agonistic effect of different compounds on GABA A1 channels

[0351]

[0352]

[0353] A: >80%, B: 60%-80% (including 60%), C: 40%-60% (including 40%), D: 20%-40% (including 20%), E: 0-20%. Table 1 shows the efficacy of the test samples at concentrations of 0.1 μM and 1 μM, that is, the ratio of the peak current generated when the test samples act on the GABA A1 receptor together with 3 μmol GABA at these two concentrations to the peak current generated when 100 μmol of GABA exists alone, multiplied by 100%.

[0354] Table 2: Electrophysiological evaluation of the agonistic effect of different compounds on GABA A1 channels

[0355] Compound <![CDATA[EC 50 (nmol)]]> <![CDATA[E max > KH002 463.9 74.6％ KH003 478.1 72.8％ KH004 403.7 106.1％ KH022 821.1 93.1％ KH025 619.9 93.7％ Comparative compound 1261 84.2％

[0356] E max is defined as the ratio of the peak current generated when the test sample coexists with 3 μmol GABA to the peak current generated when 100 μmol of GABA exists alone, multiplied by 100%.

[0357] Table 3: Electrophysiological evaluation of the agonistic effect of different compounds on GABA A4 channels

[0358]

[0359] N / A indicates that the current response does not change with concentration.

[0360] E max is defined as the ratio of the peak current generated when the test sample exists alone and when it coexists with 10 nmol GABA to the peak current generated when 10 μmol of GABA exists alone, multiplied by 100%.

[0361] Pharmacokinetics in Brain and Plasma after Administration in Example 57

[0362] 1. Experimental Protocol

[0363] 1.1 Test Drugs

[0364] The KH022 compound of this application and the reference compound (the reference compound described in Example 56).

[0365] 1.2 Test Animals

[0366] The test animals were CD-1 mice.

[0367] 1.3 Administration

[0368] 30% SBECD in water was used as the solvent for preparing the gavage administration preparation. On the first day of the experiment, the animals in Group 1 were intraperitoneally injected with the solutions of KH022 and the reference compound, and the administration volume was 5 mL / kg; the body weight of the animals was weighed before administration, and the administration volume was calculated according to the body weight. Whole blood samples (about 0.03 mL per group) were collected at the specified time by puncturing the saphenous vein (or other suitable blood collection sites). After blood sample collection, the samples were centrifuged at 3200 g for 10 minutes at 4°C, and the supernatant plasma was aspirated and quickly placed in dry ice, and maintained at -20°C or lower for LC-MS / MS analysis. After the animals were euthanized by CO2 at each time point, the brain tissues were taken, rinsed with physiological saline and dried, and then homogenized with a homogenate of 15 mM PBS (pH 7.4):MeOH = 2:1 with a volume 4 times the brain weight (g). After homogenization, the samples were transferred to labeled centrifuge tubes and quickly placed in dry ice, and maintained at -20°C or lower for LC-MS / MS analysis.

[0369] 2. Data Analysis

[0370] The HPLC-UV method was used to determine the concentration of the administration preparation. The calibration curve included at least 6 concentration levels, and R 2 ≥0.999. The concentration of the test compound in plasma was determined by high performance liquid chromatography-tandem mass spectrometry (LC-MS / MS). The retention times of the compound and the internal standard, chromatogram collection, and chromatogram integration were processed using the software Analyst (Applied Biosystems), and the data statistics were processed using the software Watson LIMS (Thermo Fisher Scientific) or Analyst (Applied Biosystems). The unit of the analyte concentration in the sample was ng / mL, and 3 significant figures were retained. All values expressed as percentages (such as % deviation and % coefficient of variation, etc.) were retained to one decimal place. WinNonlin was used TMThe non-compartmental model of the pharmacokinetic software Version 6.3 (Pharsight, Mountain View, CA) processes plasma concentrations and calculates pharmacokinetic parameters using the linear log trapezoidal method.

[0371] 3. Test Results

[0372] As shown in Figure 1 and Figure 2 shown, at the same dosing dose, compared with the control (the substance could no longer be detected in the brain at 8 h), the compound KH022 had a slower clearance rate in the mouse brain, and the content in the mouse brain at 2 h was 2.5 times that of the control.

[0373] 57.3 Mouse Anti-Anxiety Test

[0374] 57.3.1 Light-Dark Box Shuttle Experiment: The mouse light-dark shuttle box experiment is designed based on the fact that mice like to move in the dark box, but the exploratory habit of animals prompts them to try to explore the light box, and the bright light stimulation in the light box inhibits the exploratory activities of animals, thus making the animals show states similar to neurosis such as anxiety and avoidance of the dark. The anti-anxiety effect can increase the retention time of experimental animals in the light box, and can evaluate the activity of animals, thereby reflecting the anti-anxiety efficacy of drugs. The experimental animals included in the study were randomly divided into 5 groups according to body weight, including a negative control group and two different dose groups (1, 3 mg / kg) of the test compound (KH004 or KH022), with 10 animals in each group, all male. The animals were all pre-treated with the corresponding dose of the test article for 5 consecutive days, and behavioral tests were performed 30 min after the last dose. The retention time in the light box was selected as the main index to evaluate the anti-anxiety effects of KH004 and KH022.

[0375] The test results are shown in Table 4:

[0376] Table 4: Effects of KH004 and KH022 on the Retention Time of Experimental Animals in the Light Box

[0377]

[0378] Note: The data in the table are all expressed as mean ± standard deviation (Mean ± SD). Statistical analysis was performed on 10 animals in each test group. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 represents the statistical analysis results of different test groups and the animals in the negative control group.

[0379] 57.3.2 Elevated Plus-Maze Experiment: The elevated plus-maze experiment utilizes the conflict state formed by the exploratory characteristics of animals in a new environment and the fear of the high and open arms. After the experimental animals are placed in the maze, they will actively explore the open arms, but at the same time, they are afraid of the high and open environment in the open arms, thus forming anxiety-like symptoms. Anxiolytics can relieve the anxiety conflict state of animals in this experimental system. The experimental animals were randomly divided into 4 groups according to body weight, including a negative control group and two different dose groups (1 mg / kg, 3 mg / kg) of the test compounds (KH004 or KH022). Each group had 10 animals, all male. The animals were all pre-treated with the corresponding dose of the test article for 5 consecutive days. Behavioral tests were performed 30 min after the last administration. The percentage of the latency of the number of times the animals entered the open arms (Latency) was used as the main index to evaluate the anxiolytic effects of KH004 and KH022. The experimental results are shown in Table 5.

[0380] Table 5: Effects of Different Doses of KH004 and KH022 on the Retention Time of Mice in the Open Arms

[0381]

[0382] Note: The data in the table are all expressed as mean ± standard deviation (Mean ± SD). Statistical analysis was performed on 10 animals in each test group. ** P < 0.01, *** P < 0.001, **** P < 0.0001 represents the results of the statistical analysis of the animals in different test groups and the negative control group. Experimental conclusion: KH004 and KH022 significantly increased the percentage of the retention time of animals in the open arms, showing anxiolytic effects.

[0383] 57.3.3 Experiment on the Side Effects of Animals during Administration

[0384] Since drugs acting on GABA(A) will produce behavioral manifestations similar to anesthesia (side effects), the side effects of animals during administration were observed during the elevated plus-maze experiment. There were 10 mice in each group, and three behavioral manifestations were observed: gait, activity ability, and prone behavior. "+" represents the severity, and three "+" represent the most severe, with the recovery time to normal movement > 60 minutes. The positive drug was the control compound described in Example 56. The experimental results are shown in Table 6 below.

[0385] As can be seen from Table 6, compared with the positive drug, the side effects of the KH004 and KH022 compounds of the present application are significantly smaller.

[0386]

Claims

1. A compound of formula I or a pharmaceutically acceptable salt thereof, characterized in that As shown by the compound of formula I: wherein R1 is hydrogen or methyl; R2 is hydrogen; R3 is methyl; L is -CH2-; n is 1; R4 is: wherein R6 is H, halogen, CN, CF3, NO2, or C1-6 alkyl or C1-6 alkoxy which is substituted or unsubstituted by halogen.

2. A pharmaceutical composition, which comprises the compound of claim 1 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

3. Use of the compound or a pharmaceutically acceptable salt thereof as described in claim 1 in the preparation of a drug for regulating GABA.

4. Use of the compound or a pharmaceutically acceptable salt thereof as described in claim 1 in the preparation of a drug for preventing or treating nervous system diseases.

5. The use according to claim 4, wherein The nervous system diseases are selected from: sleep disorders, mood disorders, schizophrenia spectrum disorders, spastic disorders, memory disorders and / or cognitive disorders, movement disorders, personality disorders, autism spectrum disorders, pain, traumatic brain injury, vascular diseases, substance use disorders and / or truncation syndrome or tinnitus.

6. The use according to claim 5, characterized in that The mood disorder is depression; wherein the depression is disruptive mood dysregulation disorder, major depressive disorder, persistent depressive disorder, premenstrual dysphoric disorder, substance- or drug-induced disorder.

7. The use according to claim 6, characterized in that The nervous system diseases are selected from mild depression, moderate depression, severe depression or postpartum depression.

8. The pharmaceutical composition according to claim 2, wherein The pharmaceutical composition can be administered orally, subcutaneously, intravenously or intramuscularly.

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