STEROID COMPOUNDS, THEIR USE AND PREPARATION METHODS - Patent application

Abstract

To provide a steroid compound that is expected to effectively treat mental and neurological diseases and offer superior active efficacy, pharmacokinetic (PK) performance, oral bioavailability, stability, safety, clearance rate, and / or metabolic performance and the like, uses thereof and preparation methods therefor.SOLUTION: The present invention provides a pharmaceutical composition comprising a compound represented by formula (II) with a specific substituent, or a pharmaceutically acceptable salt thereof.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to the field of pharmaceuticals, in particular to steroid compounds, their uses and methods for their preparation. [Background technology]

[0002] Neuropsychiatric disorders, including anxiety disorders, depression, and schizophrenia, are the leading cause of illness in China, accounting for approximately 20% of all illnesses (Wang Juncheng et al., China Health Service Management, 2009(5): 348-350). With the relentless modernization of society, accelerating work schedules, and increasing lifestyle demands, the number of patients with various neuropsychiatric disorders has increased dramatically, and the progression of symptoms has accelerated significantly, making the development, research, and production of new drugs to treat these disorders more urgent. Meanwhile, clinical data suggest that many patients suffer from multiple neuropsychiatric disorders simultaneously. The high co-morbidity of neuropsychiatric disorders poses greater challenges for clinical treatment.

[0003] In animals, GABA (gamma-aminobutyric acid) (γ-aminobutyric acid) is found exclusively in neural tissue, and immunological studies have shown that the highest concentrations of GABA are found in the substantia nigra. GABA is an important inhibitory neurotransmitter and has been extensively studied. GABA is involved in various metabolic activities and has high physiological activity. The GABA system is the major inhibitory signaling pathway in the brain and central nervous system and is of great significance for central nervous system function. GABA(A) receptors (GABAARs) are ion receptors and ligand-gated ion channels. Its endogenous ligand, γ-aminobutyric acid (GABA), the main inhibitory neurotransmitter, is found in the central nervous system. After activation of GABA(A) receptors, extracellular Cl2 is released. - is selectively transported through the pore of the GABA receptor. -GABAergic pathways exit neuronal cells when their internal potential is lower than the resting potential and enter neuronal cells when their internal potential is higher than the resting potential (i.e., -75 mV). This effectively reduces the likelihood of action potentials and inhibits neurotransmission. Anxiety and depression have a high co-morbidity rate and are thought to overlap and co-occur. Anxiety and depression are also important psychiatric symptoms of schizophrenia. Research findings have confirmed the important role of the GABAergic system and GABA(A) receptors in the etiology and pathogenesis of these three disorders, suggesting that pathophysiological processes related to the GABAergic system may be one of the determinants of all three disorders. The GABAergic system and GABA(A) receptors have been shown to be involved in the pathological processes of anxiety, depression, and schizophrenia at molecular, preclinical, and clinical levels, and GABA(A) receptors have long been used as important drug targets in the treatment of these disorders.

[0004] In addition to binding to GABA itself and forming receptor complexes (GRCs) to alter brain excitability, clinical trials have confirmed that GABA(A) receptors can also bind to other small molecules, such as barbiturates (trade name: secobarbital sodium) and benzodiazepines (trade name: diazepam). These drugs each exert their therapeutic effects by binding to specific allosteric sites on the GABA(A) receptor. Furthermore, studies have shown that the GABA(A) receptor contains a unique binding site for steroid compounds (Lan, NC et al., Neurochem. Res. 16: 347-356 (1991)).

[0005] Neuroactive steroids can be endogenously produced.The most potent endogenous neuroactive steroids are 3α-hydroxy-5-reduced pregnan-20-one and 3α-21-dihydroxy-5-reduced pregnan-20-one, which are the metabolites of hormonal steroids progesterone and deoxycorticosterone, respectively.The ability of these steroid metabolites to change brain excitability was recognized in 1986 (Majewska, MD et al., Science 232: 1004-1007 (1986); Harrison, NL et al., J. Pharmacol. Exp. Ther.241:346-353 (1987)). The ovarian hormone progesterone and its metabolites have been demonstrated to have profound effects on brain excitability (Backstrom, T. et al, Acta Obstet. Gynecol. Scand. Suppl. 130: 19-24 (1985); Pfaff, DW and McEwen, BS, 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)). Concentrations of progesterone and its metabolites vary depending on the phase of the menstrual cycle. It is well documented that concentrations of progesterone and its metabolites decrease before the onset of menstruation. It is also well documented that certain physical symptoms recur monthly before the onset of menstruation. These symptoms have come to be associated with premenstrual syndrome (PMS) and include stress (tension), anxiety, and migraines (Dalton, K., Premenstrual Syndrome and Progesterone Therapy, 2nd edition, Chicago Yearbook, Chicago (1984)). Subjects with PMS have symptoms that recur monthly, are present before menstruation, and are not present after menstruation.

[0006] Other studies have found that decreased progesterone levels are associated with increased seizure frequency in female epileptic patients (Laidlaw, J., Lancet, 1235-1237 (1956)). A more direct correlation has been observed with decreased progesterone metabolites (Rosciszewska et al., J. Neurol. Neurosurg. Psych. 49:47-51 (1986)). Also, in subjects with primary generalized petit mal seizure epilepsy, the temporal incidence of seizures correlates with the incidence of premenstrual syndrome symptoms (Backstrom, T. et al., J. Psychosom. Obstet. Gynaecol. 2:8-20 (1983)). The steroid deoxycorticosterone has been found to be effective in treating subjects with epileptic seizures that correlate with the menstrual cycle (Aird, R.B. and Gordan, G, J. Amer. Med. Soc. 145:715-719 (1951)).

[0007] Another syndrome associated with low progesterone levels is postpartum depression (PPD). Immediately after childbirth, progesterone levels drop dramatically, leading to the onset of PPD. PPD symptoms range from mild depression to psychosis requiring hospitalization. PPD is also associated with severe anxiety and irritability. PPD is not amenable to traditional antidepressant treatment, and women experiencing PPD show an increased incidence of PMS (Dalton, K., Premenstrual Syndrome and Progesterone Therapy, 2nd edition, Chicago Yearbook, Chicago (1984)). Collectively, these observations imply that progesterone and deoxycorticosterone, and more specifically their metabolites, play an important role in the homeostatic regulation of brain excitability, which is manifested by increased seizure activity or symptoms associated with menstrual-related epilepsy, PMS, and PPD.

[0008] Many studies have focused on the GABA(A) receptor to obtain drugs that can effectively treat related diseases. CN103958540A, CN10533928A, etc. have disclosed a series of neurosteroid compounds for the treatment of neuropsychiatric disorders. GABA(A) is a subtype of the gamma-aminobutyric acid receptor (GABAR), which has three subtypes: GABA(A), GABA(B), and GABA(C). GABA(A) is a chloride ion channel.

[0009] GABA(A) is composed of five subunits (polypeptide chains) that form a heteropentameric structure. Two pairs of the five subunits are identical, so a specific GABA(A) receptor has three types of subunits, commonly known as α, β, and γ. Each subunit has several different subtypes: α has α1, α2, α3, α4, α5, and α6; β has β1, β2, and β3; and γ has γ1, γ2, γ3, and γ4. Furthermore, δ, ε, and ρ 1~3 There are subunits such as , θ, π, and can form pentamers with α and β.

[0010] Statistics show that there are a total of 23 GABA(A) receptors with different subunit combinations. The specific isoform of a receptor is related to its distribution in the brain. For example, GABA(A) receptors containing the gamma subunit are primarily located inside the synapse, while GABA(A) receptors containing the delta subunit are primarily located outside the synapse (extrasynaptic). (Activation of GABA(A) Receptors: Views from Outside the Synaptic Cleft, Neuron, Vol 56, 2007, 763-770); one study (Endogenous neurosteroids regulate GABA(A) receptors through two discrete transmembrane sites: Nature. Vol 444. 23 November 2006. 486-489) showed that endogenous steroids such as tetrahydrodeoxycorticosterone (THDOC) at high concentrations (micromolar levels) inhibit the activation of synaptic GABA receptors α1β2γ2 (hereafter referred to as GABA receptors). A1 We showed that it directly activates the GABA receptor α1β2γ2, while at low concentrations (submicromolar levels) it enhances the GABA receptor α1β2γ2.

[0011] [ka]

[0012] However, extrasynaptic receptor subtypes (such as α4β3δ, hereafter referred to as GABA A4 There have been no reports of steroid compounds that can directly activate the steroid hormone α-glucanase (hereinafter referred to as α-glucanase). Summary of the Invention

[0013] One of the objectives of the present invention is to provide steroid compounds that can more effectively treat neurological disorders. The compounds of the present invention are expected to provide good efficacy, pharmacokinetic (PK) properties, oral bioavailability, formulation feasibility, stability, safety, clearance, and / or metabolism, and to reduce side effects such as sensory anesthesia and sedation.

[0014] With the above objects in mind, the present invention provides a compound of formula (I), or a pharmaceutically acceptable salt thereof:

[0015] [ka]

[0016] During the ceremony, R1 is hydrogen, substituted or unsubstituted C1-6 alkyl, substituted or unsubstituted C1-6 alkoxy, substituted or unsubstituted C2-6 alkenyl, substituted or unsubstituted C2-6 alkynyl, or substituted or unsubstituted C3-6 carbocyclyl.

[0017] R2 is hydrogen, halogen, substituted or unsubstituted C1-6 alkyl, substituted or unsubstituted C1-6 alkoxy, substituted or unsubstituted C2-6 alkenyl, substituted or unsubstituted C2-6 alkynyl, substituted or unsubstituted C3-6 carbocyclyl.

[0018] R3 is hydrogen, unsubstituted C1-6 alkyl, or -CH2OR a where R a is hydrogen, substituted or unsubstituted C1-6 alkyl, substituted or unsubstituted C1-6 alkoxy, substituted or unsubstituted C2-6 alkenyl, substituted or unsubstituted C2-6 alkynyl, substituted or unsubstituted C3-6 carbocyclyl.

[0019] L is —C(Rb)(Rb)—, where each Rb is independently hydrogen or C1-C6 alkyl, and n is an integer of 0-3.

[0020] R4 is halogen, or substituted or unsubstituted heteroaryl or heterocyclyl.

[0021] Further preferred compounds are R1 is hydrogen, substituted or unsubstituted C1-6 alkyl; R2 is hydrogen, substituted or unsubstituted C1-6 alkyl; R3 is hydrogen, unsubstituted C1-6 alkyl; Rb is hydrogen and n is an integer from 1 to 2; R4 is optionally 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 heteroaryl substituted with N(Re)(Rf); Each Rd is independently hydrogen or C1-C6 alkyl; Each of Re and Rf is independently hydrogen, C1-C6 alkyl, carbocyclyl, heterocyclyl, aryl, or heteroaryl.

[0022] Further preferred compounds are 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 heterocyclyl, each heteroatom being oxygen, nitrogen, or sulfur, and the bicyclic rings may be spiro or fused.

[0023] Further preferred compounds are R4 is a 5- or 6-membered heteroaryl containing 2 to 4 nitrogen atoms, optionally substituted with cyano, nitro, hydroxy, halo, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl.

[0024] Or a compound of formula II:

[0025] [ka]

[0026] During the ceremony, R1 is hydrogen or a halogen-substituted or unsubstituted C1-3 alkyl group; R2 is hydrogen, halogen, a halogen-substituted or unsubstituted C1-6 alkyl group, or a halogen-substituted or unsubstituted C1-6 alkoxy group; X is CH, N, O, or S; R5 is selected from the group consisting of:

[0027] [ka]

[0028] During the ceremony, R6 is H, halogen, CN, CF3, NO2, halogen-substituted or unsubstituted C1-6 alkyl, or halogen-substituted or unsubstituted C1-6 alkoxy.

[0029] Alternatively, preferred compounds are selected from the following:

[0030] [ka] JPEG0007775397000006.jpg35169

[0031] [ka]

[0032] [ka]

[0033] The present invention also provides pharmaceutical compositions comprising any of the above compounds, or pharmaceutically acceptable salts thereof, or a pharmaceutically acceptable excipient.

[0034] The present invention also provides compounds described herein that can act as GABA modulators, such as by positively or negatively affecting GABA(A) receptors. As modulators of central nervous system (CNS) excitability, such compounds are expected to have CNS activity by virtue of their ability to modulate GABA(A) receptors.

[0035] The present invention further provides the use of the compounds or pharmaceutical compositions described herein in the prevention or treatment of neurological disorders. The neurological disorders are preferably selected from sleep disorders, mood disorders, schizophrenia spectrum disorders, spastic disorders, memory and / or cognitive disorders, movement disorders, personality disorders, autism spectrum disorders, pain, traumatic brain injury, vascular diseases, substance abuse disorders and / or withdrawal syndromes, or tinnitus; the mood disorders are depression, including major mood disorders, severe depressive disorders, persistent depressive disorders, premenstrual syndrome, substance- or drug-induced disorders, physical illness disorders, other specified depressive disorders, and unspecified depressive disorders, preferably mild depression, moderate depression, severe depression, or postpartum depression. The compounds or pharmaceutical compositions can be administered orally, subcutaneously, intravenously, or intramuscularly.

[0036] [Definition] [Chemical definition] The compounds described herein may contain one or more asymmetric centers and therefore may exist in various isomeric forms, such as enantiomers and / or diastereomers. For example, the compounds described herein may be in the form of individual enantiomers, diastereomers, or geometric isomers, or may be in the form of a mixture of stereoisomers. Such mixtures include racemic mixtures and mixtures enriched in one or more stereoisomers. Isomers can be isolated from mixtures by methods known to those skilled in the art, including chiral high-pressure liquid chromatography (HPLC) and the formation and crystallization of chiral salts; or preferred isomers can be prepared by asymmetric synthesis.

[0037] The following terms are intended to have the following meanings provided therewith and are useful for understanding the description and intended scope of the present invention. When describing the present invention, it can include compounds, pharmaceutical compositions containing the compounds, and compounds and compositions of test methods. For definitions of terms included in the present invention, reference can be made to the description below, and any of the moieties defined below may be substituted with many substituents. Corresponding definitions are within their scope listed below, including such substituted moieties. Unless otherwise specified, the term "substituted" is as defined below.

[0038] "Alkyl" refers to a straight-chain or branched saturated hydrocarbon group having 1 to 20 carbon atoms ("C1-20 alkyl"). In some embodiments, an alkyl group has 1 to 12 carbon atoms ("C1-12 alkyl"). In some embodiments, an alkyl group has 1 to 10 carbon atoms ("C1-10 alkyl"). In some embodiments, an alkyl group has 1 to 9 carbon atoms ("C1-9 alkyl"). In some embodiments, an alkyl group has 1 to 8 carbon atoms ("C1-8 alkyl"). In some embodiments, an alkyl group has 1 to 7 carbon atoms ("C1-7 alkyl"). In some embodiments, an alkyl group has 1 to 6 carbon atoms ("C1-6 alkyl", also referred to herein as "lower alkyl"). In some embodiments, an alkyl group has 1 to 5 carbon atoms ("C1-5 alkyl"). In some embodiments, an alkyl group has 1 to 4 carbon atoms ("C1-4 alkyl"). In some embodiments, an alkyl group has 1 to 3 carbon atoms ("C1-3 alkyl"). In some embodiments, an alkyl group has 1 to 2 carbon atoms ("C1-2 alkyl"). In some embodiments, an alkyl group has 1 carbon atom ("C1 alkyl"). In some embodiments, an alkyl group has 2 to 6 carbon atoms ("C2-6 alkyl"). Examples of C1-6 alkyl groups include methyl (C1), ethyl (C2), n-propyl (C3), isopropyl (C3), n-butyl (C4), tert-butyl (C4), sec-butyl (C4), isobutyl (C4), n-pentyl (C5), 3-pentanyl (C5), neopentyl (C5), 3-methyl-2-butanyl (C5), tertiary amyl (C5), and n-hexyl (C6). Further examples of alkyl groups include n-heptyl (C7), n-octyl (C8), and the like. Unless otherwise specified, each example alkyl group is independently optionally substituted, i.e., unsubstituted (an "unsubstituted alkyl") or substituted (a "substituted alkyl") with one or more substituents, e.g., 1 to 5 substituents, 1 to 3 substituents, or 1 substituent.In certain embodiments, the alkyl group is an unsubstituted C1-10 alkyl (eg, -CH3). In certain embodiments, the alkyl group is a substituted C1-10 alkyl.

[0039] "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 certain embodiments, the alkenyl does not contain any triple bonds. In some embodiments, an alkenyl group has 2 to 10 carbon atoms ("C2-10 alkenyl"). In some embodiments, an alkenyl group has 2 to 9 carbon atoms ("C2-9 alkenyl"). In some embodiments, an alkenyl group has 2 to 8 carbon atoms ("C2-8 alkenyl"). In some embodiments, an alkenyl group has 2 to 7 carbon atoms ("C2-7 alkenyl"). In some embodiments, an alkenyl group has 2 to 6 carbon atoms ("C2-6 alkenyl"). In some embodiments, an alkenyl group has 2 to 5 carbon atoms ("C2-5 alkenyl"). In some embodiments, an alkenyl group has 2 to 4 carbon atoms ("C2-4 alkenyl"). In some embodiments, an alkenyl group has 2 to 3 carbon atoms ("C2-3 alkenyl"). In some embodiments, an alkenyl group has 2 carbon atoms ("C2 alkenyl"). The one or more carbon-carbon double bonds can be internal (e.g., 2-butenyl) or terminal (e.g., 1-butenyl). Examples of C2-4 alkenyl groups include ethenyl (C2), 1-propenyl (C3), 2-propenyl (C3), 1-butenyl (C4), 2-butenyl (C4), butadienyl (C4), and the like. Examples of C2-6 alkenyl groups include the C2-4 alkenyl groups mentioned above, as well as pentenyl (C5), pentadienyl (C5), hexenyl (C6), etc. Further examples of alkenyl include heptenyl (C7), octenyl (C8), octatrienyl (C8), etc.Unless otherwise specified, each exemplary alkenyl group is independently optionally substituted, i.e., unsubstituted ("unsubstituted alkenyl") or substituted ("substituted alkenyl") with one or more substituents, e.g., 1 to 5 substituents, 1 to 3 substituents, or 1 substituent. In certain embodiments, an alkenyl group is an unsubstituted C2-10 alkenyl. In certain embodiments, an alkenyl group is a substituted C2-10 alkenyl.

[0040] "Alkynyl" refers to a straight-chain or branched hydrocarbon group having 2 to 20 carbon atoms, one or more carbon-carbon triple bonds (e.g., 1, 2, 3, or 4 carbon-carbon triple bonds), and optionally one or more carbon-carbon double bonds (e.g., 1, 2, 3, or 4 carbon-carbon double bonds) ("C2-20 alkynyl"). In certain embodiments, the alkynyl does not contain any double bonds. In some embodiments, an alkynyl group has 2 to 10 carbon atoms ("C2-10 alkynyl"). In some embodiments, an alkynyl group has 2 to 9 carbon atoms ("C2-9 alkynyl"). In some embodiments, an alkynyl group has 2 to 8 carbon atoms ("C2-8 alkynyl"). In some embodiments, an alkynyl group has 2 to 7 carbon atoms ("C2-7 alkynyl"). In some embodiments, an alkynyl group has 2 to 6 carbon atoms ("C2-6 alkynyl"). In some embodiments, an alkynyl group has 2 to 5 carbon atoms ("C2-5 alkynyl"). In some embodiments, an alkynyl group has 2 to 4 carbon atoms ("C2-4 alkynyl"). In some embodiments, an alkynyl group has 2 to 3 carbon atoms ("C2-3 alkynyl"). In some embodiments, an alkynyl group has 2 carbon atoms ("C2 alkynyl"). The one or more carbon-carbon triple bonds can be internal (e.g., 2-butynyl) or terminal (e.g., 1-butynyl). Examples of C2-4 alkynyl groups 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 alkenyl groups include the C2-4 alkynyl groups mentioned above, as well as pentynyl (C5), hexynyl (C6), etc. Further examples of alkynyl groups include heptynyl (C7), octynyl (C8), etc. Unless otherwise specified, each example alkynyl group is independently optionally substituted, i.e., unsubstituted (an "unsubstituted alkynyl") or substituted (a "substituted alkynyl") with one or more substituents, e.g., 1 to 5 substituents, 1 to 3 substituents, or 1 substituent.In certain embodiments, the alkynyl group is unsubstituted C2_10 alkynyl. In certain embodiments, the alkynyl group is substituted C2_10 alkynyl.

[0041] "Aryl" refers to 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 the cyclic arrangement) having 6 to 14 ring carbon atoms and zero heteroatoms in the aromatic ring system ("C6-14 aryl"). In some embodiments, an aryl group has a ring of 6 carbon atoms ("C6 aryl"; e.g., phenyl). In some embodiments, an aryl group has 10 ring carbon atoms ("C10 aryl"; e.g., naphthyl, such as 1-naphthyl and 2-naphthyl). In some embodiments, an aryl group has 14 ring carbon atoms ("C14 aryl"; e.g., anthracyl). "Aryl" also includes ring systems in which an aryl ring is fused with one or more carbocyclyl or heterocyclyl groups, as defined above, where the radical or point of attachment is on the aryl ring, and in such cases, the number of carbon atoms continues to represent the number of carbon atoms in the aryl ring system. Typical aryl groups include, but are not limited to, groups derived from aceanthrylene, acenaphthylene, acephenanthrylene, anthracene, azulene, benzene, chrysene, coronene, fluoranthene, fluorene, hexacene, hexaphene, hexalene, as-indacene, s-indacene, indane, indene, naphthalene, octacene, octaphene, octalene, ovalene, penta-2,4-diene, pentacene, pentalene, pentaphene, perylene, phenalene, phenanthrene, picene, pleiadene, pyrene, pyranthrene, rubicene, triphenylene, and trinaphthalene. Particular aryl groups include phenyl, naphthyl, indenyl, and tetrahydronaphthyl. Unless otherwise specified, each exemplary aryl group is independently optionally substituted, i.e., unsubstituted ("unsubstituted aryl") or substituted ("substituted aryl") with one or more substituents. In certain embodiments, the aryl group is unsubstituted C6-14 aryl. In certain embodiments, the aryl group is substituted C6-14 aryl.

[0042] "Heteroaryl" refers to a radical of a 5-10-membered monocyclic or bicyclic 4n+2 aromatic ring system (e.g., having 6 or 10 π-electrons shared in the cyclic arrangement) having ring carbon atoms and 1-4 heteroatoms in the aromatic ring system ("5-10-membered heteroaryl"). Here, each heteroatom is independently selected from nitrogen, oxygen, and sulfur. In heteroaryl groups containing one or more nitrogen atoms, the point of attachment can be a carbon atom or a nitrogen atom, if valence allows. Heteroaryl bicyclic ring systems can contain one or more heteroatoms in one or both rings. "Heteroaryl" includes ring systems in which a heteroaryl ring, as defined above, is fused with one or more carbocyclyl or heterocyclyl 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 a heteroaryl ring, as defined above, is fused with one or more aryl groups. Here, the point of attachment is on either the aryl ring or the heteroaryl ring; in such cases, the number of ring members refers to the number of ring members in the fused (aryl / heteroaryl) ring system. Bicyclic heteroaryl groups in which one ring does not contain a heteroatom (e.g., indolyl, quinolinyl, carbazolyl, etc.) can have the point of attachment on either ring, i.e., the ring containing the heteroatom (e.g., 2-indolyl) or the ring containing no heteroatoms (e.g., 5-indolyl). In some embodiments, the heteroaryl group is a 5- to 10-membered aromatic ring system ("5- to 10-membered heteroaryl") having ring carbon atoms and 1 to 4 ring heteroatoms in the aromatic ring system, where each heteroatom is independently selected from nitrogen, oxygen, and sulfur. In some embodiments, the heteroaryl group is a 5- to 8-membered aromatic ring system ("5- to 8-membered heteroaryl") having ring carbon atoms and 1 to 4 ring heteroatoms in the aromatic ring system, where each heteroatom is independently selected from nitrogen, oxygen, and sulfur. In some embodiments, the heteroaryl group is a 5-6 membered aromatic ring system having ring carbon atoms and 1-4 ring heteroatoms in the aromatic ring system (a "5-6 membered heteroaryl").wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur. In some embodiments, the 5- to 6-membered heteroaryl has 1 to 3 ring heteroatoms selected from nitrogen, oxygen, and sulfur. In some embodiments, the 5- to 6-membered heteroaryl has 1 to 2 ring heteroatoms selected from nitrogen, oxygen, and sulfur. In some embodiments, the 5- to 6-membered heteroaryl has 1 ring heteroatom selected from nitrogen, oxygen, and sulfur. Unless otherwise specified, each exemplary heteroaryl group is independently optionally substituted, i.e., unsubstituted ("unsubstituted heteroaryl") or substituted ("substituted heteroaryl") with one or more substituents. In certain embodiments, the heteroaryl group is an unsubstituted 5- to 14-membered heteroaryl. In certain embodiments, the heteroaryl group is a substituted 5- to 14-membered heteroaryl. Exemplary 5-membered heteroaryl groups containing 1 heteroatom include, but are not limited to, pyrrolyl, furanyl, and thiophenyl. Exemplary 5-membered heteroaryl groups containing two heteroatoms include, but are not limited to, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, and isothiazolyl. Exemplary 5-membered heteroaryl groups containing three heteroatoms include, but are not limited to, triazolyl, oxadiazolyl, and thiadiazolyl. Exemplary 5-membered heteroaryl groups containing four heteroatoms include, but are not limited to, tetrazolyl. Exemplary 6-membered heteroaryl groups containing one heteroatom include, but are not limited to, pyridinyl. Exemplary 6-membered heteroaryl groups containing two heteroatoms include, but are not limited to, pyridazinyl, pyrimidinyl, and pyrazinyl. Exemplary 6-membered heteroaryl groups containing three or four heteroatoms include, but are not limited to, triazinyl and tetrazinyl, respectively. Exemplary 7-membered heteroaryl groups containing one heteroatom include, but are not limited to, azepinyl, oxepinyl, and thiepinyl.Exemplary 5,6-bicyclic heteroaryl groups include, but are not limited to, indolyl, isoindolyl, indazolyl, benzotriazolyl, benzothiophenyl, isobenzothiophenyl, benzofuranyl, benzisofuranyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl, benzoxadiazolyl, benzothiazolyl, benzisothiazolyl, benzothiadiazolyl, indolizinyl, and purinyl. Exemplary 6,6-bicyclic heteroaryl groups include, but are not limited to, naphthyridinyl, pteridinyl, quinolinyl, isoquinolinyl, cinnolinyl, quinoxalinyl, phthalazinyl, and quinazolinyl.

[0043] "Carbocyclyl" or "carbocyclic" refers to the radical of a non-aromatic cyclic hydrocarbon group having 3 to 10 ring carbon atoms ("C3-10 carbocyclyl") and zero heteroatoms in the non-aromatic ring system. In some embodiments, a carbocyclyl group has 3 to 8 ring carbon atoms ("C3-8 carbocyclyl"). In some embodiments, a carbocyclyl group has 3 to 6 ring carbon atoms ("C3-6 carbocyclyl"). In some embodiments, a carbocyclyl group has 3 to 6 ring carbon atoms ("C3-6 carbocyclyl"). In some embodiments, a carbocyclyl group has 5 to 10 ring carbon atoms ("C5-10 carbocyclyl"). Exemplary C3-6 carbocyclyl 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), etc. Exemplary C3-8 carbocyclyl groups include, but are not limited to, the above C3-6 carbocyclic groups as well as cycloheptyl (C7), cycloheptenyl (C7), cycloheptadienyl (C7), cycloheptatrienyl (C7), cyclooctyl (C8), cyclooctenyl (C8), bicyclo[2.2.1]heptyl (C7), bicyclo[2.2.2]octyl (C8), etc. Exemplary C3-10 carbocyclyl groups include, but are not limited to, the C3-8 carbocyclyl groups described above, as well as 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 examples above, in some embodiments, a carbocyclyl group is a carbocyclyl group containing a monocyclic ring ("monocyclic carbocyclyl") or a fused, bridged, or spiro ring system, such as a bicyclic ring system ("bicyclic carbocyclyl"). A carbocyclyl group can be a saturated or partially unsaturated carbocyclyl group."Carbocyclyl" also includes ring systems in which a carbocyclic ring as described above is fused to one or more aryl or heteroaryl groups, where the point of attachment is on the carbocyclic ring, and in such cases the number of carbons continues to refer to the number of carbons in the carbocyclic ring system. Unless otherwise specified, each carbocyclyl group is independently optionally substituted, i.e., unsubstituted ("unsubstituted carbocyclyl") or substituted ("substituted carbocyclyl") with one or more substituents. In some embodiments, a carbocyclyl is an unsubstituted C3-10 carbocyclyl. In some embodiments, a carbocyclyl is a substituted C3-10 carbocyclyl. In some embodiments, a "carbocyclyl" is a monocyclic saturated carbocyclyl having 3 to 10 ring carbon atoms ("C3-10 cycloalkyl"). In some embodiments, a cycloalkyl group has 3 to 8 ring carbon atoms ("C3-8 cycloalkyl"). In some embodiments, a cycloalkyl group has 3 to 6 ring carbon atoms ("C cycloalkyl"). In some embodiments, a cycloalkyl group has 5 to 6 ring carbon atoms ("C cycloalkyl"). In some embodiments, a cycloalkyl group has 5 to 10 ring carbon atoms ("C cycloalkyl"). Examples of C cycloalkyl groups include cyclopentyl (C) and cyclohexyl (C). Examples of C cycloalkyl groups include the C cycloalkyl groups described above, as well as cyclopropyl (C) and cyclobutyl (C). Examples of C cycloalkyl groups include the C cycloalkyl groups described above, as well as cycloheptyl (C) and cyclooctyl (C). Unless otherwise specified, each example cycloalkyl group is independently unsubstituted ("unsubstituted cycloalkyl") or substituted ("substituted cycloalkyl") with one or more substituents. In some embodiments, a cycloalkyl is an unsubstituted C cycloalkyl. In some embodiments, the cycloalkyl is a substituted C3_10 cycloalkyl.

[0044] "Heterocyclyl" or "heterocyclic" refers to a radical of a 3- to 10-membered non-aromatic ring system having ring carbon atoms and 1 to 4 ring heteroatoms ("3- to 10-membered heterocyclyl"), where each heteroatom is independently selected from nitrogen, oxygen, sulfur, boron, phosphorus, and silicon. In heterocyclic groups containing one or more nitrogen atoms, the point of attachment can be at a carbon atom or a nitrogen atom, if valence allows. Heterocyclyl groups can be either monocyclic ("monocyclic heterocyclyl") or fused, bridged, or spiro ring systems, such as bicyclic systems ("bicyclic heterocyclyl"). Heterocyclyl groups can be saturated or partially unsaturated. Heterocyclyl bicyclic ring systems can contain one or more heteroatoms in one or both rings. "Heterocyclyl" also includes ring systems in which a heterocyclyl ring, as defined above, is fused to one or more carbocyclyl groups, with the point of attachment being on the carbocyclyl or heterocyclyl ring, or in which a heterocyclyl ring, as defined above, is fused to one or more aryl or heteroaryl groups, with the point of attachment being on the heterocyclyl ring. In such cases, the number of ring members continues to refer to the number of ring members in the heterocyclyl ring system. Unless otherwise specified, each heterocyclyl instance is independently optionally substituted, i.e., unsubstituted ("unsubstituted heterocyclyl") or substituted ("substituted heterocyclyl") with one or more substituents. In certain embodiments, a heterocyclyl group is an unsubstituted 3- to 10-membered heterocyclyl. In certain embodiments, a heterocyclyl group is a substituted 3- to 10-membered heterocyclyl. In some embodiments, a heterocyclyl group is a 5- to 10-membered non-aromatic ring system having ring carbon atoms and 1-4 ring heteroatoms ("5- to 10-membered heterocyclyl"), where each heteroatom is independently selected from nitrogen, oxygen, sulfur, boron, phosphorus, and silicon. In some embodiments, a heterocyclyl group is a 5- to 8-membered non-aromatic ring system having ring carbon atoms and 1-4 ring heteroatoms ("5- to 8-membered heterocyclyl"), where each heteroatom is independently selected from nitrogen, oxygen, and sulfur.In some embodiments, a heterocyclyl group is a 5- to 6-membered non-aromatic ring system ("5- to 6-membered heterocyclyl") having ring carbon atoms and 1 to 4 ring heteroatoms, where each heteroatom is independently selected from nitrogen, oxygen, and sulfur. In some embodiments, a 5- to 6-membered heterocyclyl has 1 to 3 ring heteroatoms selected from nitrogen, oxygen, and sulfur. In some embodiments, a 5- to 6-membered heterocyclyl has 1 to 2 ring heteroatoms selected from nitrogen, oxygen, and sulfur. In some embodiments, a 5- to 6-membered heterocyclyl has 1 ring heteroatom selected from nitrogen, oxygen, and sulfur.

[0045] "Hetero," when used to describe a compound or a group present on a compound, means that one or more carbon atoms in the compound or group are replaced with a nitrogen, oxygen, or sulfur heteroatom. Hetero can apply to any of the above hydrocarbyls: for example, alkyl, such as heteroalkyl; cycloalkyl, such as heterocyclyl; aryl, such as heteroaryl; cycloalkenyl, such as cycloheteroalkenyl; and the hydrocarbyl has 1 to 5 heteroatoms, especially 1 to 3 heteroatoms.

[0046] "Alkoxy" refers to the group -OR, where R is substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted carbocyclyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl. Particular alkoxy groups are methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, tert-butoxy, sec-butoxy, n-pentoxy, n-hexoxy, and 1,2-dimethylbutoxy. Particular alkoxy groups are lower alkoxy, i.e., those having 1 to 6 carbon atoms. More particular alkoxy groups have 1 to 4 carbon atoms. Exemplary substituted alkoxy groups include, but are not limited to, -O-(CH2) t (C6~C 10aryl), -O-(CH2) t (5-10 membered heteroaryl), -O-(CH2) t (C3~C 10 cycloalkyl), and -O-(CH2) t (4- to 10-membered heterocyclyl); where t is an integer from 0 to 4. Optionally substituted groups include any aryl, heteroaryl, cycloalkyl, or heterocyclyl group present, which may itself be substituted with an unsubstituted C1-C4 alkyl group, halo, an unsubstituted C1-C4 alkoxy group, an unsubstituted C1-C4 haloalkyl group, an unsubstituted C1-C4 hydroxyalkyl group, or an unsubstituted C1-C4 haloalkoxy group, or a hydroxy group.

[0047] "Carboxy" refers to -C(O)OH.

[0048] "Cyano" refers to -CN.

[0049] "Halo" or "halogen" refers to fluorine (F), chlorine (Cl), bromine (Br), and iodine (I). In certain embodiments, a halo group is either a fluorine or chlorine atom.

[0050] "Hydroxy" refers to --OH.

[0051] "Nitro" refers to -NO2.

[0052] [Other definitions] The term "pharmaceutically acceptable salt" refers to those salts that, within the scope of sound medical judgment, are suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response, and the like, commensurate with a reasonable benefit / risk ratio. Pharmaceutically acceptable salts are known in the art. For example, Berge et al. describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences (1977) 66:1-19. Pharmaceutically acceptable salts of the compounds of the present invention include those derived from suitable inorganic and organic acids and bases. Examples of pharmaceutically acceptable non-toxic acid addition salts are salts of amino groups. Such salts are formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, and perchloric acid, or with organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid, or malonic acid, or by using other methods used in the art, such as ion exchange. Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecyl sulfate, ethanesulfonate, formate, fumarate, glucoheptanoate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, and 2-hydroxy-ethanesulfonate. Pharmaceutically acceptable salts include, but are not limited to, lactobionate, lactate, laurate, laurate sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate, and the like. Pharmaceutically acceptable salts derived from appropriate bases include alkali metal, alkaline earth metal, ammonium, and N(C1-4 alkyl)4 salts.Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, etc. Further pharmaceutically acceptable salts include non-toxic ammonium, quaternary ammonium, and amine cations, formed where appropriate with counterions such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, lower alkylsulfonates, and arylsulfonates. [Brief explanation of the drawings]

[0053] [Figure 1] FIG. 1 is a graph showing the time course of changes in plasma and brain controls. [Figure 2] FIG. 2 is a graph showing the time course of KH022 in plasma and brain. DETAILED DESCRIPTION OF THE INVENTION

[0054] In order to better illustrate the technical concept of the present invention, the present invention provides some compound synthesis or biological examples, but does not further limit the protection scope of the present invention.

[0055] 〔material and method〕 The compounds provided herein can be prepared from readily available starting materials using the following general methods and procedures. Under typical or preferred process conditions (i.e., reaction temperature, time, molar ratio of reactants, solvent, pressure, etc.), it is understood that other process conditions can also be used unless otherwise specified. Optimal reaction conditions may vary depending on the specific reactants or solvents used. However, such conditions can be determined by one skilled in the art based on routine optimization.

[0056] Additionally, it will be apparent to one skilled in the art that conventional protecting groups may be required to prevent undesired reactions of certain functional groups. It is known in the art how to select an appropriate protecting group for a particular functional group, as well as suitable conditions for protection and deprotection.

[0057] The compounds provided herein can be isolated and purified by known standard methods, including, but not limited to, recrystallization, column chromatography, HPLC, or supercritical fluid chromatography (SFC). Details of the preparation of representative pyrazoles herein are provided in the following reaction schemes.

[0058] The compounds provided herein can be prepared according to methods described in the art using the appropriate reagents, starting materials and purification methods known to those skilled in the art.

[0059] Example 1: Synthesis of compound KH001

[0060] [ka]

[0061] Compound KHC-2: KCN (600.0 g, 9.2 mol) and acetic acid (640 mL) were added to a solution of compound KHC-1 (100.0 g, 0.3 mol) in methanol (4 L), and the reaction mixture was stirred at room temperature (RT) overnight. The reaction mixture was poured into water and extracted with ethyl acetate (EA). The organic phase was dried over anhydrous sodium sulfate and concentrated under vacuum to obtain the crude product, which was dissolved in acetic acid (2.5 L). Platinum dioxide (24.0 g, 0.1 mol) was added to the reaction mixture, and hydrogenation was carried out using H2 gas. When TLC showed the reaction was complete, the reaction mixture was filtered, and the filtrate was concentrated to approximately 200 mL and diluted with ice water (2.4 L). An aqueous solution of NaNO2 (46.0 g, 0.7 mol) was slowly added dropwise to the mixture, and the mixture was stirred at room temperature for 15 hours and extracted with EA. The combined organic layers were washed with brine, dried, concentrated, and purified by silica gel column flash chromatography to give KHC-2 (57.3 g, 55% yield).

[0062] Compound KHC-3: Ethynylmagnesium bromide (395 mL, 0.5 M) was added dropwise to a solution of compound KHC-2 (57.3 g, 164.5 mmol) in THF (1 L) at −78° C. under a N atmosphere, then gradually warmed to RT and stirred at RT overnight. Saturated aqueous NH4Cl was slowly added to the mixture, followed by extraction with EA. The organic phase was washed with brine, dried over anhydrous Na2SO4, and concentrated. The resulting crude product was purified by silica gel column flash chromatography to give compound KHC-3 (33.7 g, 55% yield).

[0063] 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 heated to reflux. After the reaction was completed, the mixture was cooled to RT, filtered, and washed with EA. The filtrate was neutralized with 40% aqueous NaOH solution, and the organic layer was washed with saturated brine, dried over anhydrous NaSO, and concentrated. The crude product was purified by silica gel column flash chromatography to give compound KHC-4 (23.6 g, 70% yield).

[0064] Compound KHC-5: Compound KHC-4 (23.6 g, 63.3 mmol) and palladium black (1.2 g) in THF (200 mL) were stirred at RT overnight and then hydrogenated with H gas. The mixture was filtered through a pad of Celite, washed with EA, and the filtrate was concentrated in vacuo to give the crude compound. The crude compound was recrystallized from acetone to give compound KHC-5 (19.0 g, 80% yield).

[0065] Compound KH001: Aqueous NaOH solution (50 mL, 1 M) was slowly added to a solution of compound KHC-5 (19.0 g, 50.8 mmol) in THF (200 ml) and stirred overnight at RT. The pH of the reaction mixture was adjusted with dilute hydrochloric acid, followed by extraction with EA. The organic phase was washed with brine, dried over anhydrous Na2SO4, and concentrated to give the crude product. The crude product was purified by silica gel column flash chromatography to give compound KH001 (13.5 g, 80% yield). 1 H NMR(400MHz,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).

[0066] Example 2: Synthesis of compound KH002

[0067] [ka]

[0068] Compound KHC-6: Compound KH001 (1.5 g, 4.5 mmol) synthesized in Example 1 was dissolved in CHOH (40 mL) until the solution became clear, and then cooled to 5 °C in ice water. Bromine (1.5 g, 9.2 mmol) was then added dropwise to the cooled solution. The ice bath was removed, and the reaction was carried out at RT for 4 h, during which the reaction solution changed from orange to pale yellow. When TLC monitoring of the reaction showed the reaction was complete, water (30 mL) was added to the reaction solution, and extraction was carried out with EA (2 × 35 mL). The combined organic layer was washed with saturated brine and dried over anhydrous NaSO. The solvent was distilled under reduced pressure to obtain a pale yellow crude solid. This crude solid was purified by silica gel column flash chromatography (eluent: CHCl) to obtain a white solid (1.0 g, 56.7% yield).

[0069] Compound KH002: Compound KHC-6 (80 mg, 0.2 mmol) was dissolved in THF (5 mL) until the solution became clear, followed by the addition of K2CO3 (83 mg, 0.6 mmol) and 4-cyanopyrazole (56 mg, 0.6 mmol). The reaction was carried out at room temperature overnight. After TLC monitoring of the reaction showed that the reaction was complete, the reaction mixture was washed with 10 mL of water and extracted with EA (2 x 20 mL). The combined organic layer was washed with saturated brine and dried over anhydrous Na2SO4. The solvent was distilled under reduced pressure to obtain a pale yellow crude solid. The crude solid was purified by silica gel column flash chromatography (eluent: CHCl2 / CH3OH = 100:1) and then recrystallized (EA / PE = 1:10) to obtain a white solid (36 mg, 42.5% yield). 1H 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).

[0070] Example 3: Synthesis of compound KH003

[0071] [ka]

[0072] Compound KH003: Compound KHC-6 (50 mg, 0.1 mmol) was dissolved in THF (5 mL) until the solution became clear, and then K2CO3 (50 mg, 0.3 mmol) and 1H-triazole (25 mg, 0.3 mmol) were added. The reaction was carried out at room temperature overnight. After TLC monitoring of the reaction showed that the reaction was complete, the reaction mixture was washed with 10 mL of water and extracted with EA (2 × 20 mL). The combined organic layer was washed with saturated brine and dried over anhydrous Na2SO4. The solvent was distilled under reduced pressure to obtain a pale yellow crude solid. The crude solid was purified by silica gel column flash chromatography (eluent: CHCl2 / CH3OH = 150:1) and then recrystallized (EA / PE = 1:10) to obtain a white solid (17 mg, 35.5% yield). 1 H 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).

[0073] Example 4: Synthesis of compounds KH004 and KH005

[0074] [ka]

[0075] Compounds KH004 and KH005: Compound KHC-6 (103 mg, 0.2 mmol) was dissolved in CHCN (15 mL) until the solution became clear, and then CsCO (244 mg, 0.7 mmol) and 1H-tetrazolium (53 mg, 0.7 mmol) were added. The reaction was carried out at room temperature. When TLC monitoring of the reaction showed that the reaction was complete, the reaction mixture was washed with 10 mL of water and extracted with EA (2 × 25 mL). The combined organic layer was washed with saturated brine and dried over anhydrous NaSO. The solvent was distilled under reduced pressure to give a pale yellow crude solid. The crude solid was purified by silica gel column flash chromatography (eluent: CHCl / CHOH = 150:1 to 90:1) to give two compounds. These two compounds were each recrystallized (EA / PE=1:10) to give white solid 1 (24 mg, 24.0% yield) and white solid 2 (36 mg, 36.0% yield). 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(KH₀O₅): 1 H 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).

[0076] [Example 5: Synthesis of compound KH006]

[0077]

change

[0078] Compound KH006: Compound KHC-6 (50 mg, 0.1 mmol) was dissolved in CHCN (8 mL) until the solution became clear, and then CsCO (117 mg, 0.3 mmol) and 1H-pyrazole (24 mg, 0.3 mmol) were added. The reaction was carried out at room temperature overnight. When TLC monitoring of the reaction showed that the reaction was complete, the reaction mixture was washed with 10 mL of water and extracted with EA (2 × 20 mL). The combined organic layer was washed with saturated brine and dried over anhydrous NaSO. The solvent was distilled under reduced pressure to give a pale yellow crude solid. The crude solid was purified by flash chromatography on a silica gel column (eluent: CHCl / CHOH = 100:1) and recrystallized (EA / PE = 1:10) to give a white solid (24 mg, 50.2% yield). 1 H 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).

[0079] Example 6: Synthesis of compound KH007

[0080] [ka]

[0081] Compound KH007: Compound KHC-6 (150 mg, 0.3 mmol) was dissolved in THF (10 mL), followed by the addition of K2CO3 (152 mg, 1.1 mmol) and 1H-imidazole (75 mg, 1.1 mmol). The reaction was carried out at room temperature overnight. When TLC monitoring of the reaction showed the reaction was complete, water (10 mL) was added to the reaction solution, and extraction was carried out with EA (2 × 20 mL). The combined organic layer was washed with saturated brine and dried over anhydrous Na2SO4. The solvent was distilled under reduced pressure to obtain a pale yellow crude solid. The crude solid was purified by silica gel column flash chromatography (eluent: CHCl2 / CH3OH = 30:1) and then recrystallized (EA) to obtain a white solid (35 mg, 24.1% yield). 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).

[0082] Example 7: Synthesis of compound KH008

[0083] [ka]

[0084] Compound KH008: Compound KHC-6 (100 mg, 0.2 mmol) was dissolved in THF (10 mL), followed by the addition of K2CO3 (97 mg, 0.7 mmol) and 3-cyanopyrazole (68 mg, 0.7 mmol). The reaction was carried out at 70 °C for 2 hours or more. When TLC monitoring of the reaction showed that the reaction was complete, water (10 mL) was added to the reaction solution, and extraction was carried out with EA (2 × 20 mL). The combined organic layer was washed with saturated brine and dried over anhydrous Na2SO4. The solvent was distilled under reduced pressure to obtain a pale yellow crude solid. The crude solid was purified by TLC (eluent: PE / acetone = 3:1) and then recrystallized (CH3CN) to obtain a white solid (20 mg, 19.4% yield). 1 H 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).

[0085] Example 8: Synthesis of compound KH009

[0086] [ka]

[0087] Compound KH009: Compound KHC-6 (50 mg, 0.1 mmol) was thoroughly dissolved in CHCN (5 mL), and then CsCO (120 mg, 0.3 mmol) and 1H-indole (68 mg, 0.7 mmol) were added. The reaction was carried out at room temperature overnight. When TLC monitoring of the reaction showed that the reaction was complete, water (10 mL) was added to the reaction solution and extracted with EA (2 × 20 mL). The combined organic layer was washed with saturated brine and dried over anhydrous NaSO. The solvent was distilled under reduced pressure to obtain a pale yellow crude solid. The crude solid was purified by TLC (eluent: CHCl / EA = 10:1) to obtain a pale yellow solid (34 mg, 62.4% yield). 1 H 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).

[0088] Example 9: Synthesis of compound KH010

[0089] [ka]

[0090] Compound KH010: Compound KHC-6 (100 mg, 0.2 mmol) was dissolved in CH3CN (5 mL), followed by the addition of K2CO3 (101 mg, 0.7 mmol) and 4-(trifluoromethyl)-1H-pyrazole (35 mg, 0.2 mmol). The reaction was carried out at 50 °C for 2 hours or more. When TLC monitoring of the reaction showed the reaction was complete, water (10 mL) was added to the reaction solution, and extraction was carried out with EA (2 × 20 mL). The combined organic layer was washed with saturated brine and dried over anhydrous Na2SO4. The solvent was distilled under reduced pressure to give a pale yellow crude solid. The crude solid was purified by silica gel column flash chromatography (eluent: PE / acetone = 7:1) and then recrystallized (CH3CN) to give a white solid (61 mg, 53.1% yield). 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).

[0091] Example 10: Synthesis of compound KH011

[0092] [ka]

[0093] Compound KH011: Compound KHC-6 (50 mg, 0.1 mmol) was dissolved in CHCN (5 mL), and then CsCO (120 mg, 0.3 mmol) and 1H-indazole (43 mg, 0.3 mmol) were added. The reaction was carried out at room temperature overnight. When TLC monitoring of the reaction showed that the reaction was complete, water (10 mL) was added to the reaction solution and extracted with EA (2 × 20 mL). The combined organic layer was washed with saturated brine and dried over anhydrous NaSO. The solvent was distilled under reduced pressure to obtain a pale yellow crude solid. The crude solid was purified by TLC (eluent: PE / acetone = 3:1) and then recrystallized (CHCN) to obtain a white solid (20 mg, 36.6% yield). 1 H 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).

[0094] Example 11: Synthesis of compound KH012

[0095] [ka]

[0096] Compound KH012: Compound KHC-6 (50 mg, 0.1 mmol) was dissolved in CHCN (5 mL), and then CsCO (120 mg, 0.3 mmol) and 1H-purine (44 mg, 0.3 mmol) were added. The reaction was carried out at room temperature overnight. When TLC monitoring of the reaction showed that the reaction was complete, water (10 mL) was added to the reaction solution, and the mixture was extracted with EA (2 × 20 mL). The combined organic layer was washed with saturated brine and dried over anhydrous NaSO. The solvent was distilled under reduced pressure to obtain the crude product as a pale yellow solid. The crude product was purified by TLC (eluent: CHCl / CHOH = 20:1) and then recrystallized (CHCN) to obtain a white solid (5 mg, 9.1% yield). 1 H 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).

[0097] Example 12: Synthesis of compound KH013

[0098] [ka]

[0099] Compound KH013: Compound KHC-6 (80 mg, 0.2 mmol) was dissolved in CHCN (8 mL), followed by the addition of CsCO (196 mg, 0.6 mmol) and 4-chloropyrazole (62 mg, 0.6 mmol). The reaction was carried out at room temperature overnight. When TLC monitoring of the reaction showed the reaction was complete, 15 mL of water was added to the reaction mixture, and the mixture was extracted with EA (2 × 30 mL). The combined organic layer was washed with saturated brine and dried over anhydrous NaSO. The solvent was distilled under reduced pressure to give a pale yellow crude solid. The crude solid was purified by silica gel column flash chromatography (eluent: CHCl / CHOH = 100:1) and then recrystallized (EA / PE = 1:10) to give a white solid (28 mg, 32.3% yield). 1 H NMR (400MHz, CDCl3) δ 7.46 (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).

[0100] Example 13: Synthesis of compound KH014

[0101] [ka]

[0102] Compound KH014: Compound KHC-6 (60 mg, 0.15 mmol) was dissolved in DMF (5 mL), and then NaOH (1 mL, 1.0 mmol) was added. The reaction was carried out at room temperature overnight. When TLC monitoring of the reaction showed that the reaction was complete, 10 mL of water was added to the reaction mixture, and the mixture was extracted with EA (2 × 20 mL). The combined organic layer was washed with saturated NaCl and dried over anhydrous NaSO. The solvent was distilled under reduced pressure to obtain the crude product. The crude product was purified by TLC (eluent: CH2Cl2 / CH3OH = 50:1) to obtain a white solid (12 mg, yield 22.92%). 1 H NMR (400 MHz, DMSO-d6) δ 4.94 - 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).

[0103] Example 14: Synthesis of compound KH015

[0104] [ka]

[0105] Compound KH015: Compound KHC-6 (60 mg, 0.15 mmol) was dissolved in CHCN (5 mL), and then CsCO (98 mg, 0.30 mmol) and 6-chloropurine (46 mg, 0.30 mmol) were added. The reaction was carried out at room temperature overnight. When TLC monitoring of the reaction showed that the reaction was complete, 10 mL of water was added to the reaction mixture, and the mixture was extracted with EA (2 × 20 mL). The combined organic layer was washed with saturated NaCl and dried over anhydrous NaSO. The solvent was distilled under reduced pressure to obtain the crude product. The crude product was purified by TLC (eluent: CHCl / CHOH = 40:1) to obtain a white solid (39 mg, yield 53.61%). 1H 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) Example 15: Synthesis of compound KH016

[0106] [ka]

[0107] Compound KH016: KHC-6 (60 mg, 0.15 mmol) was dissolved in 5 mL of CH3CN, followed by the addition of Cs2CO3 (98 mg, 0.30 mmol) and 2-mercaptothiophene (17 mg, 0.30 mmol). The reaction was carried out at room temperature overnight. When TLC monitoring of the reaction showed that the reaction was complete, 10 mL of water was added to the reaction mixture, and the mixture was extracted with EA (2 × 20 mL). The combined organic layer was washed with saturated NaCl and dried over anhydrous Na2SO4. The solvent was distilled under reduced pressure to obtain the crude product. The crude product was purified by TLC (eluent: CH2Cl2 / CH3OH = 40:1) to obtain a white solid (19 mg, 28.34% yield). 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).

[0108] Example 16: Synthesis of compound KH017

[0109] [ka]

[0110] Compound KH017: KHC-6 (60 mg, 0.15 mmol) was dissolved in 5 mL of CH3CN, followed by the addition of Cs2CO3 (98 mg, 0.30 mmol) and 6-methoxypurine (45 mg, 0.30 mmol). The reaction was carried out at room temperature overnight. When TLC monitoring of the reaction showed that the reaction was complete, 10 mL of water was added to the reaction mixture, and the mixture was extracted with EA (2 × 20 mL). The combined organic layer was washed with saturated NaCl and dried over anhydrous Na2SO4. The solvent was distilled under reduced pressure to obtain the crude product. The crude product was purified by TLC (eluent: CHCl2 / CH3OH = 40:1) to obtain a white solid (46 mg, yield 63.76%). 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).

[0111] Example 17: Synthesis of compounds KH018 and KH019

[0112] [ka]

[0113] Compounds KH018 and KH019: KHC-6 (60 mg, 0.15 mmol) was dissolved in 5 mL of CH3CN, followed by the addition of Cs2CO3 (98 mg, 0.30 mmol) and 5-methyl-1H-tetrazole (25 mg, 0.30 mmol). The reaction was carried out at room temperature overnight. When TLC monitoring of the reaction showed that the reaction was complete, 10 mL of water was added to the reaction mixture, and the mixture was extracted with EA (2 × 20 mL). The combined organic layer was washed with saturated NaCl and dried over anhydrous Na2SO4. The solvent was distilled under reduced pressure to obtain the crude product. The crude product was purified by TLC (eluent: CHCl2 / CH3OH = 40:1) to obtain KH6070053-1 (13 mg, 20.88%) as a white solid, and KH6070053-2 (27 mg, 43.37%) as a white solid, respectively. 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 - 0.76 (m, 27H), 0.74 (s, 3H). 1 H NMR (400 MHz, CDCl3) δ 5.15 (dd, 2H), 4.06 (t, 1H), 2.43 (s, 3H), 2.39 (dd, 1H), 1.95 - 0.78 (m, 27H), 0.75 (s, 3H).

[0114] Example 18: Synthesis of compound KH020

[0115] [ka]

[0116] Compound KH020: KHC-6 (60 mg, 0.15 mmol) was dissolved in 5 mL of CH3CN, followed by the addition of Cs2CO3 (98 mg, 0.30 mmol) and 2-oxa-6-aza-spiro[3,3]heptane (30 mg, 0.30 mmol). The reaction was carried out at room temperature overnight. When TLC monitoring of the reaction showed that the reaction was complete, 10 mL of water was added to the reaction mixture, and the mixture was extracted with EA (2 × 20 mL). The combined organic layer was washed with saturated NaCl and dried over anhydrous Na2SO4. The solvent was distilled under reduced pressure to obtain the crude product. The crude product was purified by TLC (eluent: CHCl2 / CH3OH = 40:1) to obtain a white solid (21 mg, 32.56% yield). 1 H 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).

[0117] Example 19: Synthesis of compound KH021

[0118] [ka]

[0119] Compound KHC-7: KH001 (0.70 g, 2.11 mmol) was dissolved in 100 mL of CHCl, and DMP (1.78 g, 4.20 mmol) was added in batches. The reaction was carried out overnight at room temperature under a N atmosphere. When TLC monitoring of the reaction showed that the reaction was complete, 40 mL of water was added to the reaction mixture, and the mixture was extracted with EA (2 × 80 mL). The combined organic layer was washed with saturated NaCl and dried over anhydrous NaSO. The solvent was distilled under reduced pressure to obtain the crude product. The crude product was purified by flash chromatography on silica gel (eluent: PE / acetone = 40:1 → 30:1) to obtain KHC-7 (0.62 g, 89.47% yield) as a white solid.

[0120] Compound KH021: FeCl3 (0.61 g, 3.75 mmol) and LiCl (0.34 g, 8.07 mmol) were mixed and added to a 100 mL three-neck flask under a N2 atmosphere. Anhydrous THF (40 mL) was then added, and the mixture was stirred at RT for 5 min and cooled to -45 °C. CH3MgBr (15.20 mL, 15.20 mmol) was then added dropwise, and the mixture was stirred at -40 °C for 15 min, followed by the dropwise addition of compound KHC-7 (0.62 g, 1.88 mmol) dissolved in anhydrous THF. The reaction was heated to approximately -20 °C and stirred for 2 h. When TLC monitoring of the reaction showed the reaction was complete, the reaction solution was quenched with 40 mL of saturated aqueous NH4Cl solution, and the mixture was extracted with EA (2 * 80 mL). The combined organic layer was washed with saturated NaCl and dried over anhydrous Na2SO4. The solvent was distilled under reduced pressure to give a crude product, which was purified by flash chromatography on silica gel (eluent: PE / acetone = 40:1 to 35:1) to give a white solid KH021 (0.52 g, yield 78.50%). 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).

[0121] Example 20: Synthesis of compounds KH022 and KH023

[0122] [ka]

[0123] [ka]

[0124] Compounds KHC-8 and KHC-9: Compound KH021 (1.25 g, 3.60 mmol) was dissolved in 40 mL of methanol until clear, and then bromine (0.86 g, 5.40 mmol) was added dropwise. After 4 hours of reaction at room temperature, the reaction solution turned from orange to pale yellow, and TLC monitoring indicated that the reaction was complete. 30 mL of water was added to the reaction mixture, and the mixture was extracted with EA (2 × 65 mL). The combined organic layer was washed with saturated sodium chloride and dried over anhydrous sodium sulfate. The solvent was distilled under reduced pressure to obtain a pale yellow crude solid. The crude solid was purified by flash chromatography on silica gel (eluted with dichloromethane) to obtain white solid KHC-8 (0.93 g, 60.78%) and white solid KHC-9 (0.36 g, 19.89%).

[0125] Compounds KH022 and KH023: Compound KHC-8 (100 mg, 0.24 mmol) was dissolved in 5 mL of acetonitrile until clear, and Cs2CO3 (234 mg, 0.72 mmol) and 1H-tetrazole (50 mg, 0.72 mmol) were added. The reaction was carried out at room temperature overnight. TLC monitoring showed that the reaction was complete. 10 mL of water was added to the reaction mixture, which was then extracted with ethyl acetate (2 × 20 mL). The combined organic layer was washed with saturated sodium chloride and dried over anhydrous sodium sulfate. The solvent was distilled under reduced pressure to obtain the crude product. The product was purified using thin-layer chromatography (TLC) on a thick silica gel preparative plate (eluted with dichloromethane / methanol = 30:1) to obtain white solids KH022 (20 mg, 20.08%) and KH023 (47 mg, 47.19%), respectively. 1 H 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). 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).

[0126] Example 21: Synthesis of compound KH024

[0127] [ka]

[0128] Compound KH024: KHC-8 (60 mg, 0.14 mmol) was dissolved in 5 mL of CH3CN, followed by the addition of Cs2CO3 (91 mg, 0.28 mmol) and 3-cyanopyrazole (26 mg, 0.28 mmol). The reaction was carried out at room temperature overnight. When TLC monitoring showed the reaction was complete, 10 mL of water was added to the reaction mixture, and the mixture was extracted with EA (2 x 20 mL). The combined organic layer was washed with saturated NaCl and dried over anhydrous Na2SO4. The solvent was distilled under reduced pressure to obtain the crude product. The crude product was purified by TLC (eluent: CH2Cl2 / CH3OH = 40:1) to obtain a white solid, KH024 (16 mg, 26.09%). 1 H NMR (400 MHz, CDCl3) 7.45 (d, 1H), 6.73 (d, 1H), 5.06 (q, 2H), 2.30 (dd, 1H), 1.86 - 1.23 (m, 21H), 1.20 (s, 3H), 0.95 (s, 3H), 0.87 - 0.77 (m, 3H), 0.72 (s, 3H).

[0129] Example 22: Synthesis of compound KH025

[0130] [ka]

[0131] Compound KH025: KHC-8 (100 mg, 0.24 mmol) was dissolved in 5 mL of DMSO, and then Cs2CO3 (235 mg, 0.72 mmol) and 3-hydroxybenzisoxazole (97 mg, 0.72 mmol) were added. The reaction was carried out at room temperature overnight. When TLC monitoring showed that the reaction was complete, 10 mL of water was added to the reaction mixture, and the mixture was extracted with EA (2 × 20 mL). The combined organic layer was washed with saturated NaCl and dried over anhydrous Na2SO4. The solvent was distilled under reduced pressure to obtain the crude product. The crude product was purified by TLC (eluent: PE / acetone = 5:1) to obtain a white solid KH025 (70 mg, 60.76%). 1 H 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) Example 23: Synthesis of compound KH026

[0132] [ka]

[0133] Compound KH026: KHC-8 (53 mg, 0.12 mmol) was dissolved in 5 mL of CH3CN, followed by the addition of K2CO3 (100 mg, 0.72 mmol) and 2-mercaptobenzoxazole (24 mg, 0.16 mmol). The reaction was carried out at room temperature overnight. When TLC monitoring showed that the reaction was complete, 10 mL of water was added to the reaction mixture, and the mixture was extracted with EA (2 x 20 mL). The combined organic layer was washed with saturated NaCl and dried over anhydrous Na2SO4. The solvent was distilled under reduced pressure to obtain the crude product. The crude product was purified by TLC (eluent: PE / acetone = 6:1) to obtain a white solid KH026 (30 mg, 50.40%). 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).

[0134] Example 24: Synthesis of compound KH027

[0135] [ka]

[0136] Compound KH027: KHC-9 (110 mg, 0.22 mmol) was dissolved in 10 mL of CH3CN, followed by the addition of Cs2CO3 (215 mg, 0.66 mmol) and 4-cyanopyrazole (61 mg, 0.66 mmol). The reaction was carried out at room temperature overnight. When TLC monitoring showed the reaction was complete, 10 mL of water was added to the reaction mixture, and the mixture was extracted with EA (2 x 20 mL). The combined organic layer was washed with saturated NaCl and dried over anhydrous Na2SO4. The solvent was distilled under reduced pressure to obtain the crude product. The crude product was purified by TLC (eluent: CH2Cl2 / CH3OH = 6:1) to obtain a white solid KH027 (76 mg, 65.30%). 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).

[0137] Example 25: Synthesis of compound KH028

[0138] [ka]

[0139] Compound KH028: KHC-8 (70 mg, 0.16 mmol) was dissolved in 8 mL of DMSO, followed by the addition of CsCO (160 mg, 0.49 mmol) and 2-hydroxybenzoxazole (66 mg, 0.49 mmol). The reaction was carried out at room temperature overnight. When TLC monitoring showed the reaction was complete, 10 mL of water was added to the reaction mixture, and the mixture was extracted with EA (2 × 20 mL). The combined organic layer was washed with saturated NaCl and dried over anhydrous NaSO. The solvent was distilled under reduced pressure to obtain the crude product. The crude product was purified by TLC (eluent: CHCl / CHOH = 60:1) to obtain a white solid, KH028 (55 mg, 71.61%). 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).

[0140] Example 26: Synthesis of compound KH029

[0141] [ka]

[0142] Compound KH029: KHC-8 (60 mg, 0.14 mmol) was dissolved in 5 mL of DMSO, and then CsCO (138 mg, 0.42 mmol) and 8-methyl-4-hydroxyquinazoline (68 mg, 0.42 mmol) were added. The reaction was carried out at room temperature overnight. When TLC monitoring showed that the reaction was complete, 10 mL of water was added to the reaction mixture, and the mixture was extracted with EA (2 × 20 mL). The combined organic layer was washed with saturated NaCl and dried over anhydrous NaSO. The solvent was distilled under reduced pressure to obtain the crude product. The crude product was purified by TLC (eluent: CHCl / CHOH = 60:1) to obtain a white solid, KH029 (50 mg, 70.72%). 1 H 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).

[0143] Example 27: Synthesis of compound KH030

[0144] [ka]

[0145] Compound KH030: KHC-8 (50 mg, 0.12 mmol) was dissolved in 5 mL of DMSO, followed by the addition of CsCO (78 mg, 0.24 mmol) and 3-hydroxy-4-methyl-5-methoxy-4H-1,2,4-triazole (31 mg, 0.24 mmol). The reaction was carried out at room temperature overnight. When TLC monitoring showed the reaction was complete, 10 mL of water was added to the reaction mixture, and the mixture was extracted with EA (2 × 20 mL). The combined organic layer was washed with saturated NaCl and dried over anhydrous NaSO. The solvent was distilled under reduced pressure to obtain the crude product. The crude product was purified by TLC (eluent: PE / acetone = 4:1) to obtain a white solid, KH030 (20 mg, 35.16%). 1 H NMR (400 MHz, CDCl3) δ 4.47 (q, 2H), 3.93 (s, 3H), 3.12 (s, 3H), 2.28 (dd, 1H), 1.86 - 1.22 (m, 21H), 1.19 (s, 3H), 0.94 (s, 3H), 0.85 - 0.78 (m, 3H), 0.70 (s, 3H).

[0146] Example 28: Synthesis of compound KH031

[0147] [ka]

[0148] Compound KH031: KHC-8 (60 mg, 0.14 mmol) was dissolved in 5 mL of CH3CN, followed by the addition of K2CO3 (58 mg, 0.42 mmol) and 1-methyl-5-mercapto-1H-tetrazole (68 mg, 0.42 mmol). The reaction was carried out at room temperature. When TLC monitoring showed the reaction was complete, 10 mL of water was added to the reaction mixture, and the mixture was extracted with EA (2 x 20 mL). The combined organic layer was washed with saturated NaCl and dried over anhydrous Na2SO4. The solvent was distilled under reduced pressure to obtain the crude product. The crude product was purified by TLC (eluent: PE / acetone = 4:1) to obtain a white solid, KH031 (50 mg, 77.47%). 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).

[0149] Example 29: Synthesis of compound KH032

[0150] [ka]

[0151] Compound KH032: KHC-8 (50 mg, 0.12 mmol) was dissolved in 5 mL of DMSO, followed by the addition of K2CO3 (50 mg, 0.36 mmol) and 1-hydroxy-1H-imidazole (30 mg, 0.36 mmol). The reaction was carried out at room temperature overnight. When TLC monitoring of the reaction showed that the reaction was complete, water (10 mL) was added to the reaction mixture, and the mixture was extracted with EA (2 × 20 mL). The combined organic layer was washed with saturated NaCl and dried over anhydrous Na2SO4. The solvent was distilled under reduced pressure to obtain the crude product. The crude product was purified by TLC (eluent: PE / acetone = 2:1) to obtain a white solid, KH032 (11 mg, 21.37%). 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).

[0152] Example 30: Synthesis of compound KH033

[0153] [ka]

[0154] Compound KH033: KHC-8 (60 mg, 0.14 mmol) was dissolved in 5 mL of DMSO, followed by the addition of CsCO (138 mg, 0.42 mmol) and 2-hydroxyquinoline (61 mg, 0.42 mmol). The reaction was carried out overnight at room temperature. When TLC monitoring of the reaction showed that the reaction was complete, water (10 mL) was added to the reaction mixture, and the mixture was extracted with EA (2 × 20 mL). The combined organic layer was washed with saturated NaCl and dried over anhydrous NaSO. The solvent was distilled under reduced pressure to obtain the crude product. The crude product was purified by TLC (eluent: PE / acetone = 4:1) to obtain a white solid, KH033 (15 mg, 21.87%). 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).

[0155] Example 31: Synthesis of compound KH034

[0156] [ka]

[0157] Compound KH034: KHC-8 (60 mg, 0.14 mmol) was dissolved in 5 mL of DMSO, followed by the addition of K2CO3 (58 mg, 0.42 mmol) and 3-hydroxy-1H-1,2,4-triazole (36 mg, 0.42 mmol). The reaction was carried out at room temperature overnight. When TLC monitoring of the reaction showed that the reaction was complete, water (10 mL) was added to the reaction mixture, and the mixture was extracted with EA (2 × 20 mL). The combined organic layer was washed with saturated NaCl and dried over anhydrous Na2SO4. The solvent was distilled under reduced pressure to obtain the crude product. The crude product was purified by TLC (eluent: CHCl2 / CH3OH = 40:1) to obtain a white solid, KH034 (40 mg, 66.45%). 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 - 1.25 (m, 21H), 1.20 (s, 3H), 0.95 (s, 3H), 0.87 - 0.79 (m, 3H), 0.71 (s, 3H).

[0158] Example 32: Synthesis of compound KH035

[0159] [ka]

[0160] Compound KH035: KHC-8 (80 mg, 0.19 mmol) was dissolved in 6 mL of DMSO, followed by the addition of Cs2CO3 (184 mg, 0.56 mmol) and 8-fluoro-4-hydroxyquinazoline (92 mg, 0.56 mmol). The reaction was carried out at room temperature overnight. When TLC monitoring of the reaction showed that the reaction was complete, water (10 mL) was added to the reaction mixture, and the mixture was extracted with EA (2 × 20 mL). The combined organic layer was washed with saturated NaCl and dried over anhydrous Na2SO4. The solvent was distilled under reduced pressure to obtain the crude product. The crude product was purified by TLC (eluent: PE / acetone = 2:1) to obtain a white solid, KH035 (65 mg, 67.21%). 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).

[0161] Example 33: Synthesis of compound KH036

[0162] [ka]

[0163] Compound KH036: KHC-8 (60 mg, 0.14 mmol) was dissolved in 5 mL of DMSO, followed by the addition of K2CO3 (58 mg, 0.42 mmol) and 6-fluoropyridine[3,4-d]pyrimidin-4-one (69 mg, 0.42 mmol). The reaction was carried out overnight at room temperature. When TLC monitoring of the reaction showed that the reaction was complete, water (10 mL) was added to the reaction mixture, and the mixture was extracted with EA (2 × 20 mL). The combined organic layer was washed with saturated NaCl and dried over anhydrous Na2SO4. The solvent was distilled under reduced pressure to obtain the crude product. The crude product was purified by TLC (eluent: CHCl2 / CH3OH = 40:1) to obtain a white solid, KH036 (40 mg, 56.02%). 1 H 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).

[0164] Example 34: Synthesis of compound KH037

[0165] [ka]

[0166] Compound KH037: KHC-8 (60 mg, 0.14 mmol) was dissolved in 5 mL of CH3CN, followed by the addition of K2CO3 (58 mg, 0.42 mmol) and 4-methyl-4H-3-mercapto-1,2,4-triazole (49 mg, 0.42 mmol). The reaction was carried out at room temperature overnight. When TLC monitoring of the reaction showed that the reaction was complete, water (10 mL) was added to the reaction mixture, and the mixture was extracted with EA (2 x 20 mL). The combined organic layer was washed with saturated NaCl and dried over anhydrous Na2SO4. The solvent was distilled under reduced pressure to obtain the crude product. The crude product was purified by TLC (eluent: PE / acetone = 3:2) to obtain a white solid, KH037 (53 mg, 82.30%). 1 H 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).

[0167] Example 35: Synthesis of compound KH038

[0168] [ka]

[0169] Compound KH038: KHC-8 (60 mg, 0.14 mmol) was dissolved in 5 mL of DMSO, and then Cs2CO3 (138 mg, 0.42 mmol) and 3-hydroxyisoxazole (36 mg, 0.42 mmol) were added. The reaction was carried out at room temperature overnight. When TLC monitoring of the reaction showed that the reaction was complete, water (10 mL) was added to the reaction mixture, and the mixture was extracted with EA (2 × 20 mL). The combined organic layer was washed with saturated NaCl and dried over anhydrous Na2SO4. The solvent was distilled under reduced pressure to obtain the crude product. The crude product was purified by TLC (eluent: PE / acetone = 15:1) to obtain a white solid KH038 (39 mg, 65.00%). 1 H 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).

[0170] Example 36: Synthesis of compound KH039

[0171] [ka]

[0172] Compound KH039: KHC-8 (75 mg, 0.18 mmol) was dissolved in 5 ml of CH3CN, followed by the addition of Cs2CO3 (171 mg, 0.53 mmol) and 5-phenyltetrazole (77 mg, 0.53 mmol). The reaction was carried out at room temperature overnight. When TLC monitoring of the reaction showed that the reaction was complete, water (10 mL) was added to the reaction mixture, and the mixture was extracted with EA (2 × 20 mL). The combined organic layer was washed with saturated NaCl and dried over anhydrous Na2SO4. The solvent was distilled under reduced pressure to obtain the crude product. The crude product was purified by TLC (eluent: PE / acetone = 3:1) to obtain a white solid, KH039 (79 mg, 89.39%). 1 H 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).

[0173] Example 37: Synthesis of compound KH040

[0174] [ka]

[0175] Compound KH040: KHC-8 (60 mg, 0.14 mmol) was dissolved in 5 mL of CH3CN, followed by the addition of Cs2CO3 (91 mg, 0.28 mmol) and 4-cyanopyrazole (26 mg, 0.28 mmol). The reaction was carried out at room temperature overnight. When TLC monitoring of the reaction showed that the reaction was complete, water (10 mL) was added to the reaction mixture, and the mixture was extracted with EA (2 × 20 mL). The combined organic layer was washed with saturated NaCl and dried over anhydrous Na2SO4. The solvent was distilled under reduced pressure to obtain the crude product. The crude product was purified by TLC (eluent: CH2Cl2 / CH3OH = 40:1) to obtain a white solid, KH040 (16 mg, 26.09%). 1 H NMR (400 MHz, CDCl3) δ 7.82 (s, 1H), 7.81 (s, 1H), 5.00 (q, 2H), 2.32 (dd, 1H), 1.88 - 1.22 (m, 21H), 1.20 (s, 3H), 0.94 (s, 3H), 0.87 - 0.79 (m, 3H), 0.71 (s, 3H).

[0176] Example 38: Synthesis of compound KH041

[0177] [ka]

[0178] Compound KH041: KHC-8 (70 mg, 0.16 mmol) was dissolved in 5 mL of CH3CN, followed by the addition of Cs2CO3 (160 mg, 0.49 mmol) and 2-mercaptothiophenone (57 mg, 0.49 mmol). The reaction was carried out at room temperature overnight. When TLC monitoring of the reaction showed that the reaction was complete, water (10 mL) was added to the reaction mixture, and the mixture was extracted with EA (2 x 20 mL). The combined organic layer was washed with saturated NaCl and dried over anhydrous Na2SO4. The solvent was distilled under reduced pressure to obtain the crude product. The crude product was purified by TLC (eluent: CHCl2 / PE = 1:1) to obtain a gray solid KH041 (27 mg, 35.53%). 1 H 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).

[0179] Example 39: Synthesis of compound KH042

[0180] [ka]

[0181] Compound KH042: KHC-8 (150 mg, 0.35 mmol) was dissolved in water (15 mL) and acetone (30 mL), and then K2CO3 (49 mg, 0.35 mmol) was added. The reaction was carried out at room temperature overnight. When TLC monitoring of the reaction showed that the reaction was complete, water (10 mL) was added to the reaction mixture, and the mixture was extracted with EA (2 x 30 mL). The combined organic layer was washed with saturated NaCl and dried over anhydrous Na2SO4. The solvent was distilled under reduced pressure to obtain the crude product. The crude product was purified by TLC (eluent: CH2Cl2 / CH3OH = 50:1) to obtain a white solid KH042 (21 mg, 16.54%). 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).

[0182] Example 39: Synthesis of compounds KH043 and KH044

[0183] [ka]

[0184] Compounds KH043 and KH044: KHC-8 (60 mg, 0.14 mmol) was dissolved in 5 mL of CH3CN, followed by the addition of Cs2CO3 (91 mg, 0.28 mmol) and 1H-triazole (15 mg, 0.21 mmol). The reaction was carried out at room temperature overnight. When TLC monitoring of the reaction indicated completion of the reaction, water (10 mL) was added to the reaction mixture, and the mixture was extracted with EA (2 × 20 mL). The combined organic layer was washed with saturated NaCl and dried over anhydrous Na2SO4. The solvent was distilled under reduced pressure to obtain the crude product. The crude product was purified by TLC (eluent: CHCl2 / CH3OH = 40:1) to obtain white solid KH6070100-1 (12 mg, 20.70%) and white solid KH6070100-2 (27 mg, 46.58%), respectively. 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). 1 H 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).

[0185] Example 40: Synthesis of compound KH045 Compound KHC-11: Compound KHC-10 (150.0 g, 183.8 mmol) was dissolved in THF (500 mL), and 10% Pd / C (6.0 g) was added to the solution. The solution was then hydrogenated under H2 gas and stirred at room temperature for 24 hours, after which it was filtered. The filtrate was concentrated in vacuo to obtain a crude product. The crude product was then stirred with a solution (eluent: PE:acetone = 1:1) to obtain compound KHC-11 (150.0 g, yield 99.26%).

[0186] Compound KHC-12: KHC-11 (150.0 g, 547.4 mmol) was dissolved in 1000 mL of CHOH, and then I (14.0 g, 54.8 mmol) was added to the solution. The solution was heated to 60 °C and stirred for 12 h. The solution was then concentrated under vacuum, and the residue was purified by silica gel column flash chromatography (eluent: PE:EA = 10:1) to give a white solid, KHC-12 (130.0 g, 74.20%).

[0187] Compound KHC-13: KHC-12 (130.0 g, 406.2 mmol) was dissolved in 1000 ml of DMF, and t-BuOK (91.0 g, 812.4 mmol) and trimethyliodonium sulfide (165.8 g, 812.7 mmol) were added to the solution. The solution was stirred at room temperature under N gas for 12 hours. Subsequently, brine was added to the reaction mixture, and the mixture was extracted with EA (3 × 1 L). The organic layers were combined and concentrated under vacuum to give a white solid, KHC-13 (120.0 g, 88.50%).

[0188] Compound KHC-14: KHC-13 (120.0 g, 359.4 mmol) was dissolved in 900 mL of ethanol and 160 mL of water, and NaN (70.2 g, 1077.9 mmol) and NH Cl (67.2 g, 1257.9 mmol) were added to the solution. oThe mixture was stirred at 0°C overnight. Brine was added to the reaction mixture, and the mixture was extracted with EA (3 x 1 L). The organic layers were combined and concentrated in vacuo. The residue was purified by silica gel column flash chromatography (eluent: PE:EA = 10:1) to give a white solid KHC-14 (130.0 g, 95.59%).

[0189] Compound KHC-15: KHC-14 (125.6 g, 332.4 mmol) was dissolved in 1000 mL of CH3CN, and NaI (249.3 g, 1662.0 mmol) was added to the solution. After stirring the solution at room temperature for 30 minutes, TMSCl (144.9 g, 1329.6 mmol) was added dropwise to the reaction solution. The reaction was stirred at room temperature for 4 hours. Brine was added to the reaction mixture, and the mixture was extracted with EA (3 x 1 L). The organic layers were combined and concentrated under vacuum. The residue was purified by silica gel column flash chromatography (eluent: PE:EA = 10:1) to give a white solid, KHC-15 (72.0 g, 75.20%).

[0190] Compound KHC-16: KHC-15 (72.0 g, 250.0 mmol) was dissolved in 1000 mL of CHOH, and I (19.0 g, 75.0 mmol) was added to the solution. The reaction solution was heated to 60 °C and stirred for 12 h, then concentrated under vacuum. The residue was purified by silica gel column flash chromatography (eluent: PE:EA = 10:1) to give a white solid, KHC-16 (53.0 g, 63.50%).

[0191] Compound KHC-17: Trimethylsilylacetylene (77.8 g, 793.0 mmol) was taken up in 1000 mL of THF, and n-butyllithium solution (1.6 m, 329.0 mmol, 205.6 ml) was added at -78 °C. The reaction solution was stirred for 2 hours. Subsequently, compound KHC-16 (53.0 g, 158.6 mmol) in 400 mL of THF was added to the reaction solution. The mixture was stirred at -78 °C for 2 hours, gradually heated to 0 °C, and then poured into saturated NH4Cl solution. The mixture was extracted with EA (800 mL x 3). The organic layers were combined and concentrated under vacuum. The residue was purified by chromatography column (eluent: PE:EA = 10:1) to obtain a white solid KHC-17 (56.0 g, 81.55%).

[0192] Compound KHC-18: KHC-17 (56.0 g, 129.3 mmol) in 800 mL of THF was mixed with 1.0 M TBAF (129.3 mmol, 129.3 ml) at 0 °C, and the mixture was stirred for 1 hour. Brine was added to the reaction mixture, and the mixture was extracted with EA (3 × 1 L). The organic layers were combined and concentrated under vacuum. The residue was purified by silica gel column flash chromatography (eluent: PE:EA = 5:1) to give a white solid KHC-18 (43.0 g, 92.30%).

[0193] Compound KHC-19: KHC-18 (38.0 g, 105.6 mmol) in 800 mL of THF was mixed with 1.0 M HCl to pH = 3, and the mixture was stirred at room temperature for 12 hours. Brine (1.0 L) was added to the reaction mixture, and the mixture was extracted with EA (3 x 1 L). The organic layers were combined and concentrated in vacuo. The residue was purified by silica gel column flash chromatography (eluent: PE:EA = 5:1) to give a white solid, KHC-19 (32.0 g, 96.50%).

[0194] Compound KHC-20: Additionally, methanesulfonic acid (148.0 g, 1541.6 mmol) was mixed with 1000 mL of CHCl, and KHC-19 (32.0 g, 101.9 mmol) in CHCl (1000 mL) was added dropwise to the solution. o The mixture was stirred at RT for 2 h and cooled to room temperature. Brine (1.0 L) was added to the reaction mixture, and the mixture was extracted with CHCl (3×1.0 L). The organic layers were combined and concentrated in vacuo. The residue was purified by silica gel column flash chromatography (eluent: PE:EA=10:1) to give a white solid, KHC-20 (20.0 g, 62.50%).

[0195] Compound KHC-21: Compound KHC-20 (20.0 g, 63.7 mmol) was dissolved in 500 mL of THF, and 10% Pd / C (3.0 g) was added to the solution. The solution was then hydrogenated under H2 gas. The reaction solution was stirred at room temperature for 2 days and then filtered. The filtrate was concentrated under vacuum to obtain a crude product. The crude product was then purified by silica gel column flash chromatography (eluent: PE:EA = 15:1) to obtain a white solid, KHC-21 (17.0 g, 84.40%).

[0196] Compound KH045: In a 2000 mL three-neck flask, under N2, FeCl3 (23.0 g, 142.0 mmol) and LiCl (13.5 g, 321.4 mmol) were mixed and 800 mL of anhydrous THF was added. The mixture was stirred at room temperature for 20 minutes and then cooled to -40 °C. CH3MgBr (3 M, 190 mL, 570.0 mmol) was added dropwise to the solution, followed by -40 °C. o Stirring was continued for 30 minutes at -20 °C. KHC-21 (17.0 g, 53.8 mmol) in anhydrous THF (200 mL) was added dropwise to the above reaction solution, and then the solution was cooled to -20 °C. oThe mixture was heated to °C and stirred for 4 hours. When TLC monitoring of the reaction showed that the reaction was complete, the reaction solution was quenched with saturated NH4Cl (40 mL) and the mixture was extracted with EA (3 x 1.0 L). The combined organic layer was washed with saturated brine I and dried over anhydrous Na2SO4. Concentration under vacuum gave the crude product. The crude product was then purified by flash chromatography on silica gel (eluent: PE / EA = 15:1) to give a white solid KH045 (10.2 g, yield 57.10%). 1 H 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).

[0197] [ka]

[0198] Example 41: Synthesis of compound KH046

[0199] [ka]

[0200] Compound KHC-22 and Compound KHC-23: Compound KH045 (1.50 g, 4.50 mmol) was dissolved in 40 mL of CHOH, and Br (1.08 g, 6.75 mmol) was added dropwise to the solution. After reacting at room temperature for 4 hours, the reaction solution changed color from orange to pale yellow. When TLC monitoring of the reaction showed that the reaction was complete, water (30 mL) was added to the reaction mixture, and the mixture was extracted with EA (2 × 65 mL). The combined organic layer was washed with saturated NaCl and dried over anhydrous NaSO. The solvent was distilled under reduced pressure to obtain the crude product. The crude product was purified by flash chromatography on silica gel (eluent: CHCl) to obtain white solid KHC-22 (1.19 g, 64.34%) and white solid KHC-23 (0.33 g, 17.84%).

[0201] Compound KH046: KHC-22 (50 mg, 0.12 mmol) was dissolved in 5 mL of DMSO, followed by the addition of K2CO3 (21 mg, 0.15 mmol) and 8-fluoro-4-hydroxyquinazoline (25 mg, 0.15 mmol). The reaction was carried out overnight at room temperature. When TLC monitoring of the reaction showed that the reaction was complete, water (10 mL) was added to the reaction mixture, and the mixture was extracted with EA (2 x 20 mL). The combined organic layer was washed with saturated NaCl and dried over anhydrous Na2SO4. The solvent was distilled under reduced pressure to obtain the crude product. The crude product was purified by TLC (eluent: PE / acetone = 4:1) to obtain a white solid KH046 (33 mg, 55.56%). 1 H 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), 1.1 5 - 0.80 (m, 8H).

[0202] Example 42: Synthesis of compound KH047

[0203] [ka]

[0204] Compound KH047: KHC-22 (50 mg, 0.12 mmol) was dissolved in CH3CN (5 mL), followed by the addition of Cs2CO3 (49 mg, 0.15 mmol) and 5-mercapto-1-methyl-1H-tetrazolium (17 mg, 0.15 mmol). The reaction was carried out at room temperature overnight. When TLC monitoring of the reaction showed that the reaction was complete, water (10 mL) was added to the reaction mixture, and the mixture was extracted with EA (2 x 20 mL). The combined organic layer was washed with saturated NaCl and dried over anhydrous Na2SO4. The solvent was distilled under reduced pressure to obtain the crude product. The crude product was purified by TLC (eluent: PE / acetone = 4:1) to obtain a white solid, KH047 (51 mg, 50.12%). 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).

[0205] Example 43: Synthesis of compound KH048

[0206] [ka]

[0207] Compound KH048: KHC-22 (60 mg, 0.14 mmol) was dissolved in 5 mL of DMSO, followed by the addition of K2CO3 (58 mg, 0.42 mmol) and 6-fluoropyridine[3,4-d]pyrimidin-4-one (69 mg, 0.42 mmol). The reaction was carried out overnight at room temperature. When TLC monitoring of the reaction showed that the reaction was complete, water (10 mL) was added to the reaction mixture, and the mixture was extracted with EA (2 × 20 mL). The combined organic layer was washed with saturated NaCl and dried over anhydrous Na2SO4. The solvent was distilled under reduced pressure to obtain the crude product. The crude product was purified by TLC (eluent: PE / CH3OH = 40:1) to obtain a white solid KH048 (37 mg, 50.12%). 1 H 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).

[0208] Example 44: Synthesis of compound KH049

[0209] [ka]

[0210] Compound KH049: KHC-22 (60 mg, 0.15 mmol) was dissolved in 5 mL of CH3CN, followed by the addition of Cs2CO3 (62 mg, 0.19 mmol) and 2-mercaptobenzoxazole (29 mg, 0.19 mmol). The reaction was carried out at room temperature overnight. When TLC monitoring of the reaction showed that the reaction was complete, water (10 mL) was added to the reaction mixture, and the mixture was extracted with EA (2 x 20 mL). The combined organic layer was washed with saturated NaCl and dried over anhydrous Na2SO4. The solvent was distilled under reduced pressure to obtain the crude product. The crude product was purified by TLC (eluent: PE / acetone = 4:1) to obtain a white solid, KH049 (42 mg, 42.92%). 1 H 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).

[0211] Example 45: Synthesis of compound KH050

[0212] [ka]

[0213] Compound KH050: KHC-22 (60 mg, 0.15 mmol) was dissolved in 5 mL of CH3CN, followed by the addition of Cs2CO3 (62 mg, 0.19 mmol) and 3-mercapto-4-methyl-4H-1,2,4-triazole (22 mg, 0.19 mmol). The reaction was carried out at room temperature overnight. When TLC monitoring of the reaction showed that the reaction was complete, water (10 mL) was added to the reaction mixture, and the mixture was extracted with EA (2 × 20 mL). The combined organic layer was washed with saturated NaCl and dried over anhydrous Na2SO4. The solvent was distilled under reduced pressure to obtain the crude product. The crude product was purified by TLC (eluent: CHCl2 / CH3OH = 70:1) to obtain a white solid, KH050 (17 mg, 25.41%). 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).

[0214] Example 46: Synthesis of compound KH051

[0215] [ka]

[0216] Compound KH051: KHC-22 (60 mg, 0.15 mmol) was dissolved in 5 mL of DMSO, followed by the addition of 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. When TLC monitoring of the reaction showed that the reaction was complete, water (10 mL) was added to the reaction mixture, and the mixture was extracted with EA (2 × 20 mL). The combined organic layer was washed with saturated NaCl and dried over anhydrous Na2SO4. The solvent was distilled under reduced pressure to obtain the crude product. The crude product was purified by TLC (eluent: CHCl2 / CH3OH = 40:1) to obtain a white solid, KH051 (27 mg, 46.36%). 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 - 1.96 (m, 1H), 1.88 - 1.82 (m, 1H), 1.79 (s, 21H), 1.11 - 0.81 (m, 8H).

[0217] Example 47: Synthesis of Compound KH052 and Compound KH053

[0218] [ka]

[0219] Compound KH052 and Compound KH053: KHC-22 (100 mg, 0.24 mmol) was dissolved in 5 mL of CH3CN, followed by the addition of Cs2CO3 (234 mg, 0.72 mmol) and 1H-tetrazole (50 mg, 0.72 mmol). The reaction was carried out at room temperature overnight. When TLC monitoring of the reaction showed that the reaction was complete, water (10 mL) was added to the reaction mixture, and the mixture was extracted with EA (2 × 20 mL). The combined organic layer was washed with saturated NaCl and dried with anhydrous Na2SO4. The solvent was distilled under reduced pressure to obtain the crude product. The crude product was purified by TLC (eluent: CHCl2 / CH3OH = 30:1) to obtain white solid KH6070069-1 (19 mg, 19.74%) and white solid KH6070069-2 (44 mg, 45.72%), respectively. 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). 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).

[0220] Example 48: Synthesis of compound KH054

[0221] [ka]

[0222] Compound KH054: KHC-22 (60 mg, 0.15 mmol) was dissolved in 5 mL of CH3CN, followed by the addition of Cs2CO3 (62 mg, 0.19 mmol) and 2-hydroxyoxazole[4,5-b]pyridine (26 mg, 0.19 mmol). The reaction was carried out at room temperature overnight. When TLC monitoring of the reaction showed that the reaction was complete, water (10 mL) was added to the reaction mixture, and the mixture was extracted with EA (2 × 20 mL). The combined organic layer was washed with saturated NaCl and dried over anhydrous Na2SO4. The solvent was distilled under reduced pressure to obtain the crude product. The crude product was purified by TLC (eluent: CHCl2 / CH3OH = 70:1) to obtain a white solid, KH054 (17 mg, 42.92%). 1 H 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).

[0223] Example 49: Synthesis of compound KH055

[0224] [ka]

[0225] Compound KH055: KHC-22 (60 mg, 0.15 mmol) was dissolved in 5 mL of CH3CN, followed by the addition of Cs2CO3 (156 mg, 0.48 mmol) and 4-cyanopyrazole (45 mg, 0.48 mmol). The reaction was carried out at room temperature overnight. When TLC monitoring of the reaction showed that the reaction was complete, water (10 mL) was added to the reaction mixture, and the mixture was extracted with EA (2 × 20 mL). The combined organic layer was washed with saturated NaCl and dried over anhydrous Na2SO4. The solvent was distilled under reduced pressure to obtain the crude product. The crude product was purified by TLC (eluent: CHCl2 / CH3OH = 30:1) to obtain a white solid, KH055 (51 mg, 50.12%). 1 H 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).

[0226] Example 50: Synthesis of compound KH056

[0227] [ka]

[0228] Compound KH056: KHC-23 (40 mg, 0.10 mmol) was dissolved in 5 mL of CH3CN, followed by the addition of Cs2CO3 (198 mg, 0.3 mmol) and 4-cyanopyrazole (28 mg, 0.30 mmol). The reaction was carried out at room temperature overnight. When TLC monitoring of the reaction showed that the reaction was complete, water (10 mL) was added to the reaction mixture, and the mixture was extracted with EA (2 × 20 mL). The combined organic layer was washed with saturated NaCl and dried over anhydrous Na2SO4. The solvent was distilled under reduced pressure to obtain the crude product. The crude product was purified by TLC (eluent: CH2Cl2 / CH3OH = 30:1) to obtain a white solid, KH056 (12 mg, 23.30%). 1 H 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).

[0229] Example 51: Synthesis of compound KH057

[0230] [ka]

[0231] Compound KH057: KHC-22 (60 mg, 0.15 mmol) was dissolved in 5 mL of DMSO, followed by the addition of Cs2CO3 (143 mg, 0.44 mmol) and 2-hydroxybenzoxazole (59 mg, 0.44 mmol). The reaction was carried out overnight at room temperature. When TLC monitoring of the reaction showed that the reaction was complete, water (10 mL) was added to the reaction mixture, and the mixture was extracted with EA (2 × 20 mL). The combined organic layer was washed with saturated NaCl and dried over anhydrous Na2SO4. The solvent was distilled under reduced pressure to obtain the crude product. The crude product was purified by TLC (eluent: PE / acetone = 3:1) to obtain a white solid, KH057 (30 mg, 42.92%). 1 H 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).

[0232] Example 52: Synthesis of compound KH058

[0233] [ka]

[0234] Compound KH058: KHC-22 (80 mg, 0.19 mmol) was dissolved in 5 mL of DMSO, followed by the addition of CsCO (190 mg, 0.58 mmol) and 8-methyl-4-quinazolinone (93 mg, 0.58 mmol). The reaction was carried out overnight at room temperature. When TLC monitoring of the reaction showed that the reaction was complete, water (10 mL) was added to the reaction mixture, and the mixture was extracted with EA (2 × 20 mL). The combined organic layer was washed with saturated NaCl and dried over anhydrous NaSO. The solvent was distilled under reduced pressure to obtain the crude product. The crude product was purified by TLC (eluent: PE / acetone = 3:1) to obtain a white solid, KH058 (28 mg, 30.01%). 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).

[0235] Example 53: Synthesis of compound KH059

[0236] [ka]

[0237] Compound KH059: KHC-22 (80 mg, 0.19 mmol) was dissolved in 5 mL of DMSO, followed by the addition of CsCO (190 mg, 0.58 mmol) and 8-hydroxy-1,7-naphthyridine (85 mg, 0.58 mmol). The reaction was carried out overnight at room temperature. When TLC monitoring of the reaction showed that the reaction was complete, water (10 mL) was added to the reaction mixture, and the mixture was extracted with EA (2 × 20 mL). The combined organic layer was washed with saturated NaCl and dried over anhydrous NaSO. The solvent was distilled under reduced pressure to obtain the crude product. The crude product was purified by TLC (eluent: PE / acetone = 3:1) to obtain a white solid, KH058 (28 mg, 30.89%). 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).

[0238] Example 54: Synthesis of compound KH060

[0239] [ka]

[0240] Compound KH060: KHC-22 (100 mg, 0.24 mmol) was dissolved in 5 mL of DMSO, followed by the addition of CsCO (156 mg, 0.48 mmol) and 3-hydroxy-4-methyl-5-methoxy-4H-1,2,4-triazole (62 mg, 0.48 mmol). The reaction was carried out at room temperature overnight. When TLC monitoring of the reaction showed that the reaction was complete, water (10 mL) was added to the reaction mixture, and the mixture was extracted with EA (2 × 20 mL). The combined organic layer was washed with saturated NaCl and dried over anhydrous NaSO. The solvent was distilled under reduced pressure to obtain the crude product. The crude product was purified by TLC (eluent: PE / acetone = 4:1) to obtain a white solid, KH058 (56 mg, 50.72%). 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).

[0241] Example 55: Synthesis of compound KH061

[0242] [ka]

[0243] Compound KH061: KHC-22 (80 mg, 0.19 mmol) was dissolved in 5 mL of DMSO, followed by the addition of Cs2CO3 (156 mg, 0.48 mmol) and 2-hydroxyquinoline (84 mg, 0.58 mmol). The reaction was carried out at room temperature overnight. When TLC monitoring of the reaction showed that the reaction was complete, water (10 mL) was added to the reaction mixture, and the mixture was extracted with EA (2 × 20 mL). The combined organic layer was washed with saturated NaCl and dried over anhydrous Na2SO4. The solvent was distilled under reduced pressure to obtain the crude product. The crude product was purified by TLC (eluent: PE / acetone = 4:1) to obtain a white solid KH058 (45 mg, 49.76%). 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), 2.04 - 2.00 (m, 1H), 1.86 - 1.83 (m, 1H), 1.78 - 1.12 (m, 21H), 1.10 - 0.80 (m, 8H).

[0244] Example 56: In vitro cell viability assay The control compounds used in the present invention have the chemical structures set forth below:

[0245] [ka]

[0246] The detailed synthesis of the control compound was prepared with reference to CN105339381B.

[0247] The present invention relates to recombinant GABA A Two subunits of the receptor, namely α1β2γ2 and α4β3δ, are applied to evaluate the performance of exemplary compounds in vitro by patch clamp electrophysiology.

[0248] 56.1 Electrophysiological Test Solutions (Extracellular and intracellular fluid composition) Extracellular solution: 140mM NaCl, 5mM CsCl, 2mM CaCl2·2H2O, 1mM MgCl2·6H2O, 5mM HEPES, 10mM D-glucose, pH=7.4 (NaOH).

[0249] Intracellular solution: 130mM CsCl, 0.1mM CaCl2·H2O, 2mM MgCl2·6H2O, 1.1mM EGTA, 5mM Na2-ATP, 10mM HEPES, pH=7.2 (CsOH).

[0250] The prepared intracellular solution was dispensed into 1 mL aliquots per tube, frozen, and stored in a refrigerator at -20°C. Freshly thawed intracellular solution was used daily for experiments. All intracellular solution was consumed within 3 months.

[0251] [56.2 Cell lines with stable expression of ion channels]

[0252] [Table 1]

[0253] [56.3 Cell culture] [56.3.1 GABA A (α1β2γ2) cell culture] GABA A HEK293 cell lines stably expressing the (α1β2γ2) receptor were cultured in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum, 800 μg / mL G418, 200 μg / mL hygromycin B, and 100 μg / mL Zeocin®. Cells were grown in a humidified incubator at 37°C with 5% carbon dioxide.

[0254] Cell passage: The old medium was removed, and the cells were washed once with PBS. Subsequently, 1 mL of 0.25% trypsin-EDTA solution was added, and the cells were incubated at 37°C for 0.5 minutes. After the cells were detached from the bottom of the dish, 5 mL of complete medium preheated to 37°C was added. Cell aggregates were dissociated by gentle pipetting. The cell suspension was transferred to a sterile centrifuge tube, and then spun at 1000 rpm for 5 minutes to collect the cells. 2.5 x 10 cells were plated onto a 6 cm cell culture dish (final medium volume: 5 mL). 5 The cells were expanded or maintained by seeding.

[0255] To maintain electrophysiological performance, cell density should not exceed 80%.

[0256] Before manual patch clamp testing, cells were detached using 0.25% trypsin-EDTA solution. 3 Cells were plated on cover slips and cultured in 24-well plates (final medium volume: 500 μL) and tested by patch clamp after 18 hours.

[0257] [56.3.2 GABA A (α4β3δ) cell culture] The HEK293 cell line was cultured in DMEM containing 10% fetal bovine serum at 37°C under 5% carbon dioxide.

[0258] Cell passage: The old medium was removed, and the cells were washed once with PBS. Subsequently, 1 mL of 0.25% trypsin-EDTA solution was added to digest the cells. The culture dish was incubated at 37°C for 0.5 minutes in a preheated incubator. As soon as the cells detached, approximately 5 mL of complete medium preheated at 37°C was added. Cell aggregates were dissociated by gentle pipetting. The cell suspension was transferred to a sterile centrifuge tube and then spun at approximately 1000 rpm for 5 minutes. 2.5 x 10 cells were placed in a 6 cm dish (final medium volume: 5 mL). 5 Cells were expanded or maintained by seeding 8 × 10 cells for transfection.3 Cells were plated onto cover slips and cultured in 24-well plates (final medium volume: 5 mL).

[0259] Transfection: On day 2, GABA A4 Transfection was performed using the X-tremeGENE HP DNA Transfection Reagent Kit (α4, β3, δ plasmid ratio 1:1:1). The ratio of plasmid to transfection reagent was 1 μg:2 μL. The amount of plasmid in a 24-well plate was 0.5 μg per well, and 1 μL of transfection reagent. To prepare 12 wells, the following specific steps were taken: 600 μL of Opti-MEM was added to a sterile centrifuge tube along with 6 μg of plasmid and mixed thoroughly. Subsequently, 12 μL of transfection reagent was added. After incubation at room temperature for 15 minutes, the mixture was mixed thoroughly. Subsequently, the transfection complex was inoculated onto the cells (50 μL per well), and the transfection complex was mixed by gently swirling.

[0260] The mixture was replaced on the third day, and patch clamping was performed on the fourth day.

[0261] 56.4 Patch Clamp Testing Glass pipettes (BF 150-86-10, Sutter Instruments) were pulled using a micropipette puller (P97, Sutter Instruments). Under a microscope (IX71, Olympus), the glass pipette was manipulated using a micromanipulator (MP285, Sutter Instruments). After contacting the cell, a high seal resistance (GΩ) was achieved by applying slight suction. After achieving a high seal, fast capacitance (pF) compensation was performed to rupture the membrane. After achieving whole-cell mode, cell capacitance (pF) compensation was performed from whole-cell capacitance compensation. No leakage current subtraction was performed.

[0262] Test and control solutions were flowed via a gravity-fed solution delivery system into a recording chamber attached to the stage of an inverted microscope. Solutions were collected from the recording chamber by vacuum suction during the experiment. Each concentration was tested multiple times. All experiments were performed at room temperature.

[0263] GABA A1 The experimental protocol for whole-cell model current recording of receptors was as follows: at tight seal, cells were held at a holding potential of -70 mV, and peak currents were recorded in gap-free mode. Cells were stimulated with 3 μM GABA for active currents. After 30 seconds of test substance application, 3 μM GABA was applied to each concentration of test substance, followed by a 2-minute washout with extracellular solution. Finally, 100 μM GABA was used as a positive control. Data were collected using an EPC-10 amplifier and stored in PatchMaster (HEKA) software.

[0264] GABA A4 The experimental protocol for whole-cell model current recording of receptors was as follows: at tight seal, cells were held at a holding potential of -70 mV, and peak currents were recorded in gap-free mode. Cells were stimulated with 10 nM GABA for activation currents. After 30 s of test substance application, various concentrations of test substance were applied with 10 nM GABA, followed by a 2-minute washout with extracellular solution. Finally, 10 μM GABA was used as a positive control. Data were collected using an EPC-10 amplifier and stored in PatchMaster (HEKA) software.

[0265] [56.5 Data Quality Standards] The following criteria are used to determine whether the data are acceptable: (1) Electrode resistance<5MΩ (2) Seal resistance > 1GΩ (3) Starting access resistance <15MΩ (4) Access resistance end <15MΩ (5) There is no obvious natural current decay.

[0266] (6) When the membrane potential is -70mV, there is no obvious leakage current.

[0267] 56.6 Data Analysis GABA A1 For the detection of 3 μM GABA currents and various concentrations of test compounds, the currents were normalized to the currents at 100 μM GABA.

[0268]

number

[0269] The activation rate corresponding to each concentration was then calculated. A4 In the detection of 10 nM GABA current and various concentrations of test compound, each current was normalized to 10 μM GABA current.

[0270]

number

[0271] The activation rate corresponding to each concentration was then calculated. The mean, standard deviation, and standard error were calculated for each test group. Nonlinear regression was used to calculate the EC50 value: Y=Bottom+(Top-Bottom) / (1+10^((LogEC50-X)×HillSlope)) To fit the data to a concentration response, EC50 was the concentration giving the half-maximal effect and Hillslope was the Hill coefficient. Curve fitting and EC50 calculations were performed using Graphpad 5.0 software.

[0272] [56.7 In vitro experiment results]

[0273] [Table 2]

[0274] A:>80%, B:60%-80% (including 60%), C:40%-60% (including 40%), D:20%-40% (including 20%), E:0-20%.

[0275] Table 1 shows the potency of exemplary compounds at concentrations of 0.1 μM and 1 μM. Potency is the ratio of GABA to the peak current produced only with 100 μmol of GABA. A1 The ratio was calculated based on the peak current generated at the receptor and multiplied by 100%. A1 The peak currents produced at the receptors were measured by the exemplary compounds at each of these two concentrations, which were comparable to those of GABA at 3 μmol. A1 The mechanism was to act on the receptor.

[0276] [Table 3]

[0277] E max is the ratio of GABA to the peak current produced only by 100 μmol of GABA A1 It is defined as the ratio of the peak current produced at the GABA receptor to the peak current produced at the receptor, multiplied by 100%. A1 The peak current generated at the receptor was measured by the GABA receptor when the exemplary compound was administered with 3 μmol of GABA. A1 It acts on the receptor.

[0278] [Table 4]

[0279] N / A indicates that the current does not change with concentration.

[0280] E max is the ratio of the peak current induced by GABA to that induced by GABA alone. A1 It is defined as the ratio of the peak current produced at the GABA receptor to the peak current produced at the receptor, multiplied by 100%. A1 The peak current generated at the receptor is A1It acts on the receptor alone or together with 10 nmol of GABA.

[0281] Example 57: Brain and plasma pharmacokinetics after administration [1. Protocol] 1.1 Test drug KH022 of the present invention and the control described in Example 56 [1.2 Animals] The experimental animals were CD-1 mice.

[0282] 1.3 Administration Preparation of test compounds for intragastric administration using 30% SBECD in water. On the first day of the experiment, animals in Group 1 were intraperitoneally injected with KH022 or control solution at a dose of 5 mL / kg. Before administration, animals were weighed, and the administration volume was calculated according to body weight. Whole blood samples (approximately 0.03 mL per group) were collected at specific times via saphenous vein puncture (or other appropriate collection site). Blood samples were immediately processed to plasma by centrifugation at 3,200 g for 10 minutes at approximately 4°C, quickly placed on dry ice, and maintained below -20°C until LC / MS / MS analysis. At each time point, animals were euthanized using CO2, and brain tissue was collected, rinsed, and dried with saline. It was homogenized with 4 times the brain weight (g) of 15 mM PBS (pH 7.4):MeOH = 2:1. After homogenization, samples were transferred to labeled centrifuge tubes, quickly placed on dry ice, and stored at or below −20° C. for LC-MS / MS analysis.

[0283] 2. Data Analysis The concentration of the test compound in the dose formulation samples was determined by LC / UV, and the calibration curve included at least six calibration standards and was calculated using the R 2The correlation coefficient was ≥ 0.999. The concentrations of test compounds in biological matrices were measured using an LC / MS / MS method. The retention time of the test compound or internal standard, chromatogram acquisition, and chromatogram integration were processed by Software Analyst (Applied Biosystems), and data statistics were processed by Software Watson LIMS (Thermo Fisher Scientific) or Analyst (Applied Biosystems). The unit of analyte concentration in the sample was ng / mL, with three significant digits reserved, and all values ​​expressed as percentages (such as deviation % and coefficient of variation %) rounded to one decimal place. WinNonlin TM Plasma concentrations were treated using a non-compartmental model in Version 6.3 (Pharsight, Mountain View, CA) pharmacokinetic software, and pharmacokinetic parameters were calculated using the linear-logarithmic trapezoidal method.

[0284] [Experimental Results] As shown in Figures 1 and 2, at the same dose, KH022 was eliminated more slowly in the mouse brain than in the control (no detection of the control in the brain at 8 hours), and the content of KH022 in the mouse brain was 2.5 times that of the control at 2 hours.

[0285] 57.3 Anti-anxiety tests in mice [57.3.1 Light / Dark Swap Box] The light-dark alternating box is a standard and well-characterized assay for the anxiolytic effects of exemplary compounds. Light makes mice evasive and stressful, so mice naturally avoid the bright box. However, their exploratory behavior also encourages them to explore the bright box. An increase in the time spent in the bright box by exemplary compounds reflects the anxiolytic effects on mice. Experimental mice were randomly divided into five groups based on body weight, including a negative control group and two different dose groups (1 mg / kg and 3 mg / kg) of the test compound (KH004 or KH022). Each group contained 10 male mice. For the test intervention, the mice were administered the corresponding dose for five consecutive days, and the anxiolytic experiment was performed 30 minutes after the last administration. The retention time in the bright box was selected as the primary index for evaluating the anxiolytic effects of KH004 and KH022.

[0286] [Experimental Results]

[0287] [Table 5]

[0288] Note: All data in the table are expressed as mean ± standard deviation (mean ± SD). Statistical analysis was performed on 10 animals in each experimental group. * P<0.05, ** P<0.01, *** P<0.001, **** P<0.0001 represents a difference between experimental and negative control animals.

[0289] [57.3.2 Elevated Plus Maze (EPM) Test] The exploratory behavioral model (EPM) was used to assess general motor and anxiety-like behaviors. This model consisted of two open and two closed arms extending from a common central platform, forming an anxiety-like symptom. Anxiolytic compounds reduce the animals' anxiety-conflict state in the test system. Experimental mice were randomly divided into four groups based on body weight, including a negative control group and two different dose groups (1 mg / kg and 3 mg / kg) of the test compounds (KH004 or KH022). Each group contained 10 male mice. For the test intervention, mice were administered the corresponding dose for five consecutive days, and the anxiolytic experiment was conducted 30 minutes after the final administration. The percentage of time the mice spent in the open arms (latency time) was used as the primary index for evaluating the anxiolytic effects of KH004 and KH022. The results are shown in Table 5.

[0290] [Table 6]

[0291] Note: All data in the table are expressed as mean ± standard deviation (mean ± SD). Statistical analysis was performed on 10 animals in each experimental group. * P<0.05, ** P<0.01, *** P<0.001, **** P<0.0001 indicates differences between animals in each experimental group and the negative control group.Conclusion: KH004 and KH022 significantly increased the percentage of time spent in the open arms of the animals, demonstrating anxiolytic effects.

[0292] [57.3.3 Experiments on side effects during administration] Because drugs acting on GABA(A) receptors can cause behavioral paralysis (side effects), mice were observed for side effects during the elevated plus maze experiment. Ten mice per group were observed for three behavioral manifestations: walking, agility, and prone behavior. "+" indicates severity. Three "+" marks indicate the most severe symptoms, with the time required to resume normal behavior exceeding 60 minutes. The positive drug was the control compound described in Example 56, and the results are shown in Table 6.

[0293] KH004 and KH022 have fewer side effects compared to the positive drugs in Table 6.

[0294] [Table 7]

Claims

1. A pharmaceutical composition comprising a compound of formula (II), or a pharmaceutically acceptable salt thereof, 【Chemistry 1】 During the ceremony, R1 is hydrogen or halogen-substituted or unsubstituted C1-3 alkyl; R2 is hydrogen, halogen, halogen-unsubstituted or substituted C1-6 alkyl, or halogen-unsubstituted or substituted C1-6 alkoxy; X is CH ２ and R5 is selected from the group consisting of: 【Chemistry 2】 During the ceremony, R6 is H, halogen, CN, CF3, NO2, halogen-unsubstituted or substituted C1-6 alkyl, or halogen-unsubstituted or substituted C1-6 alkoxy; Pharmaceutical compositions.

2. 1. A pharmaceutical composition comprising a compound, or a pharmaceutically acceptable salt thereof, The compound is selected from the group consisting of: 【Transformation 3】 【Chemistry 4】 Pharmaceutical compositions.

3. 10. The pharmaceutical composition of claim 1, comprising a pharmaceutically acceptable excipient.

4. The pharmaceutical composition of claim 2, comprising a pharmaceutically acceptable excipient.

5. 5. The pharmaceutical composition of claim 4, wherein the pharmaceutical composition is administered orally, subcutaneously, intravenously, or intramuscularly.

6. Use of the pharmaceutical composition according to any one of claims 1 to 5 in the preparation of a medicament for preventing or treating a neurological disease.

7. 7. The use according to claim 6, wherein the neurological disorder is selected from the group consisting of sleep disorders, mood disorders, schizophrenia spectrum disorders, spastic disorders, memory and / or cognitive disorders, movement disorders, personality disorders, autism spectrum disorders, pain, traumatic brain injury, vascular diseases, substance abuse disorders and amputation syndromes, or tinnitus.

8. the mood disorder is depression, 8. The use of claim 7, wherein the depression is major mood disorder, severe depressive disorder, persistent depressive disorder, premenstrual syndrome, substance- or drug-induced disorder, other somatic disorder, or other specified depressive disorder and unspecified depressive disorder.

9. 7. The use of claim 6, wherein the neurological disorder is mild depression, moderate depression, severe depression, or postpartum depression.

Images