RU2826300C2 - Steroid compound, use thereof and method for preparation thereof - Google Patents

Abstract

FIELD: chemistry; pharmaceutics.

SUBSTANCE: group of inventions relates to pharmaceutical chemistry and includes individual steroid compounds specified in the claim, a pharmaceutical composition based thereon and use.

EFFECT: steroid compounds having GABA_A1 receptor agonist activity.

8 cl, 2 dwg, 6 tbl, 57 ex

Description

Field of technology

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

LEVEL OF TECHNOLOGY

[0002] Neuropsychiatric diseases, including anxiety, depression, schizophrenia and so on, rank first in the overall morbidity in China, accounting for about 20% of all diseases (Wang Juncheng et al., China Health Service Management, 2009 (5): 348-350). With the continuous social development, accelerated work rhythm and increased life pressure, the number of patients with various neuropsychiatric diseases has increased significantly and the progression of symptoms has accelerated significantly, which requires more urgent development, research and development of new drugs for the treatment of these diseases. Meanwhile, clinical data suggest that many patients suffer from various neuropsychiatric disorders at the same time. The high comorbidity of neuropsychiatric diseases makes the treatment in the clinic face great difficulties.

[0003] In animals, GABA (gamma-aminobutyric acid, γ-aminobutyric acid) is present only in nervous tissues, and immunological studies have shown that the highest concentration of GABA is in the substantia nigra. GABA is an important inhibitory neurotransmitter that has been studied extensively. It is involved in various metabolic functions and has high physiological significance. The GABA system is the main inhibitory signaling pathway in the brain and central nervous system, which is of great importance for the functioning of the central nervous system. The GABA(A) receptor (GABAAR) is an ion receptor and ligand-gated ion channel. Its endogenous ligand, γ-aminobutyric acid (GABA), is the main inhibitory neurotransmitter in the central nervous system. After activation of the GABA(A) receptor, extracellular Cl ^- can selectively pass through the pores of GABA receptors. Cl ^- exits nerve cells when the internal potential is less than the resting potential, and enters nerve cells when the internal potential is greater than the resting potential (i.e., -75 mV). It can effectively reduce the probability of action potentials and inhibit the transmission of nerve impulses. Anxiety and depression are very often comorbid, and they are thought to have overlapping and co-occurring mechanisms. Anxiety and depression are also important emotional symptoms in schizophrenia. Research findings support that the GABAergic system and the GABA(A) receptor play an important role in the etiology and pathology of these three diseases, indicating that the pathophysiological process associated with the GABAergic system may be one of the decisive factors in all three diseases. At the molecular level, preclinical and clinical levels, the GABAergic system and the GABA(A) receptor have been shown to be involved in the pathological processes of anxiety, depression and schizophrenia, and the GABA(A) receptor has long been used as an important drug target in the treatment of these diseases.

[0004] Clinical trials have confirmed that in addition to binding to GABA itself to form a receptor complex (GRC) to alter brain excitability, the GABA(A) receptor can bind to other low-molecular-weight compounds such as barbiturates (trade name secobarbital sodium), benzodiazepine (trade name diazepam), etc. These drugs bind to specific allosteric sites on the GABA(A) receptor and thus exert their therapeutic effects. Furthermore, studies have shown that the GABA(A) receptor has unique binding sites for steroid compounds (Lan, NC et al., Neurochem. Res. 16: 347-356 (1991)).

[0005] Neuroactive steroids can occur endogenously. The most potent endogenous neuroactive steroids are 3α-hydroxy-5-reduced pregnan-20-one and 3α-21-dihydroxy-5-reduced pregnan-20-one, metabolites of the hormonal steroids progesterone and deoxycorticosterone, respectively. The ability of these steroid metabolites to alter brain excitability was discovered in 1986 (Majewska, M. D. et al., Science 232: 1004-1007 (1986); Harrison, N. L. et al., J. Pharmacol. Exp. Ther, 241:346-353 (1987)). The ovarian hormone progesterone and its metabolites have been shown to have profound effects on brain excitability (Backstrom, T. et al., Acta Obstet. Gynecol. Scand. Suppl. 130: 19-24 (1985); Pfaff, D. W and McEwen, B. S., 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)). Levels of progesterone and its metabolites vary with the phases of the menstrual cycle. It is well documented that levels of progesterone and its metabolites decline before the onset of menstruation. The monthly recurrence of certain physical symptoms before the onset of menstruation has also been well documented. These symptoms, which have come to be associated with premenstrual syndrome (PMS), include stress (tension), anxiety, and migraine headaches (Dalton, K., Premenstrual Syndrome and Progesterone Therapy, 2nd edition, Chicago Yearbook, Chicago (1984)). Subjects with PMS experience symptoms that recur monthly and are present before menstruation and absent after menstruation.

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

[0007] A syndrome also associated with low progesterone levels is postpartum depression (PPD). Immediately after childbirth, progesterone levels drop sharply, resulting in PPD. Symptoms of PPD range from mild depression to psychosis requiring hospitalization. PPD is also associated with severe anxiety and irritability. PPD is not responsive to classical antidepressants, and women experiencing PPD have been shown to have an increased incidence of PMS (Dalton, K., Premenstrual Syndrome and Progesterone Therapy, 2nd edition, Chicago Yearbook, Chicago (1984)). Taken together, these observations imply a key role for progesterone and deoxycorticosterone, and in particular their metabolites, in the homeostatic regulation of brain excitability, which manifests itself as increased seizure activity or symptoms associated with catamenial epilepsy, PMS, and PDD.

[0008] In many studies to obtain drugs that can effectively treat similar diseases, GABA(A) receptors are used as a basis. CN103958540A, CN10533928A, etc. describe a series of neurological steroid compounds for the treatment of neuropsychiatric diseases. GABA(A): a subtype of γ-aminobutyric acid receptor (GABAR), GABAR occurs in three subtypes, including GABA(A), GABA(B), and GABA(C). GABA(A) is a chloride ion channel.

[0009] GABA(A) consists of five subunits (polypeptide chains) that form a heteropentameric structure. Among the five subunits, two pairs are identical, so that a specific GABA(A) receptor contains three types of subunits, commonly known as α, β, and γ. Each subunit has several different subtypes, for example, α is α ₁ , α ₂ , α ₃ , α ₄ , α ₅ , α ₆ , β is β ₁ , β ₂ , β ₃ , γ is γ ₁ , γ ₂ , γ ₃ , γ ₄ . Moreover, there are subunits such as δ, ε, ρ _1-3 , θ, π, etc., which can form pentamers with α and β.

[0010] According to statistics, there are a total of 23 GABA(A) receptors with different combinations of subunits. There is a relationship between the specific receptor isoform and the distribution area of neurons in the brain. For example, GABA(A) receptors containing the γ subunit are predominantly located inside synapses, whereas GABA(A) receptors containing the δ subunit are predominantly distributed outside synapses (extrasynaptic).

(Activation of GABA(A) Receptors: Views from Outside the Synaptic Cleft, Neuron, vol. 56, 2007, 763-770). The 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) can directly activate the GABA receptor α ₁ β ₂ γ ₂ in synapses (hereafter referred to as GABA _A1 ) at high concentrations (micromolar level), while potentiating the GABA receptor α ₁ β ₂ γ ₂ at lower concentrations (submicromolar level).

[0011]

[0012] However, there is no information on steroid compounds that can directly activate receptor subtypes outside synapses (such as α ₄ β ₃ δ, hereinafter referred to as GABA _A4 ).

BRIEF DESCRIPTION OF THE INVENTION

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

[0015] In view of the above objects, the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof:

[0016]

[0017] wherein 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;

[0018] 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;

[0019] R3 is hydrogen, unsubstituted C1-6 alkyl, or -CH ₂ OR ^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;

[0020] L is -C(Rb)(Rb)-, each Rb is independently hydrogen or C1-C6 alkyl, n is an integer from 0 to 3;

[0021] R4 is halogen or substituted or unsubstituted heteroaryl or heterocyclyl.

[0022] Further preferred are compounds wherein 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, n is an integer from 1 to 2; R4 is heteroaryl optionally substituted with cyano, nitro, hydroxy, halogen, 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) _0-2Rd , -S(O) _0-2ORd , or -S(O) _0-2N (Re)(Rf); each Rd is independently hydrogen or C1-C6 alkyl; each Re and Rf is independently hydrogen, C1-C6 alkyl, carbocyclyl, heterocyclyl, aryl, heteroaryl.

[0023] Further preferred are compounds wherein R1 is hydrogen or methyl; R2 is hydrogen; R3 is hydrogen; L is _-CH2- ; R4 is monocyclic or bicyclic heteroaryl or monocyclic or bicyclic heterocyclyl, each heteroatom is oxygen or nitrogen or sulfur, the bicyclic ring may be spiro or fused.

[0024] Further preferred are compounds wherein R4 is a five- or six-membered heteroaryl containing 2-4 nitrogen atoms and optionally containing cyano, nitro, hydroxy, halogen, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl as a substituent.

[0025] Or compounds corresponding to formula II:

[0026] (II)

[0027]

[0028] where R1 is hydrogen or a C1-3 alkyl group containing a halogen substituent or no substituents; R2 is hydrogen, a halogen, a C1-6 alkyl group containing a halogen substituent or no substituents, or a C1-6 alkoxy group containing a halogen substituent or no substituents; X is _CH2 , N, O or S; R5 is selected from the following groups:

[0029]

[0030] where R6 is H, halogen, CN, CF3, _NO2 , C1-6 alkyl, with or without halogen substituent, or C1-6 alkoxy, with or without halogen substituent.

[0031] or the preferred compound is selected from:

[0032]

[0033]

[0033]

[0034]

[0035] The present invention also provides pharmaceutical compositions comprising any of the above compounds or pharmaceutically acceptable salts of said compounds or pharmaceutically acceptable excipients.

[0036] The present invention also provides compounds described herein that can act as GABA modulators, such as positively or negatively affecting GABA(A) receptors. Such compounds, since they are modulators of central nervous system (CNS) excitability, are expected to exhibit activity in the CNS due to their ability to modulate GABA(A) receptors.

[0037] According to the present invention, there is further provided the use of the compounds described herein or the pharmaceutical compositions for the prevention or treatment of a neurological disease; wherein the neurological disease is preferably selected from a sleep disorder, a mood disorder, a schizophrenia spectrum disorder, a spastic disorder, a memory disorder and/or a cognitive disorder, a movement disorder, a personality disorder, an autism spectrum disorder, pain, traumatic brain injury, a vascular disorder, a substance abuse disorder, and/or a withdrawal syndrome or tinnitus; wherein the mood disorder is depression; wherein the depression is a disruptive mood disorder, a major depressive disorder, a persistent depressive disorder, a premenstrual dysphoria, a substance- or drug-induced disorder, a disorder due to physical illness, other specified depressive disorder and a non-specific depressive disorder, preferably mild depression, moderate depression, severe depression or postpartum depression. Severe depression or postpartum depression is preferable. And the above compounds or pharmaceutical compositions can be administered orally, subcutaneously, intravenously or intramuscularly.

[0038] Definitions

[0039] Definitions of chemical substances

[0040] The compounds described herein may contain one or more asymmetric centers and, therefore, may exist in different isomeric forms, such as enantiomers and/or diastereomers. For example, the compounds described herein may be in the form of a single enantiomer, diastereomer, or geometric isomer, or may be in the form of a mixture of stereoisomers, including racemic mixtures and mixtures enriched in one or more stereoisomers. The isomers can be separated from the mixture by methods known to those skilled in the art, including chiral high-pressure liquid chromatography (HPLC) and the formation and crystallization of chiral salts; or preferred isomers can be obtained by asymmetric synthesis.

[0041] The following terms are intended to have the following meanings as provided herein and are useful in understanding the present specification and the intended scope of the present invention. The present invention provides compounds, pharmaceutical compositions containing said compounds, and compounds and compositions of a test method. Definitions of terms used in the present invention can be found in the following description, any part defined below may contain a plurality of substituents, and the corresponding definitions are within the scope thereof provided below, including such substituted parts. Unless otherwise specified, the term "containing substituents" has the meaning provided below.

[0042] “Alkyl” refers to a straight or branched chain saturated hydrocarbon group containing from 1 to 20 carbon atoms (“C1-20 alkyl”). In some embodiments, the alkyl group contains from 1 to 12 carbon atoms (“C1-12 alkyl”). In some embodiments, the alkyl group contains from 1 to 10 carbon atoms (“C1-10 alkyl”). In some embodiments, the alkyl group contains from 1 to 9 carbon atoms (“C1-9 alkyl”). In some embodiments, the alkyl group contains from 1 to 8 carbon atoms (“C1-8 alkyl”). In some embodiments, the alkyl group contains from 1 to 7 carbon atoms (“C1-7 alkyl”). In some embodiments, the alkyl group contains from 1 to 6 carbon atoms (“C1-6 alkyl,” also referred to herein as “lower alkyl”). In some embodiments, the alkyl group contains from 1 to 5 carbon atoms (“C1-5 alkyl”). In some embodiments, the alkyl group contains from 1 to 4 carbon atoms (“C1-4 alkyl”). In some embodiments, the alkyl group contains from 1 to 3 carbon atoms (“C1-3 alkyl”). In some embodiments, the alkyl group contains from 1 to 2 carbon atoms (“C1-2 alkyl”). In some embodiments, the alkyl group contains 1 carbon atom (“C1 alkyl”). In some embodiments, the alkyl group contains from 2 to 6 carbon atoms (“C2-6 alkyl”). Examples of the C1-6 alkyl group include methyl (C1), ethyl (C2), n-propyl (C3), isopropyl (C3), n-butyl (C4), tert-butyl (C4), sec-butyl (C4), iso-butyl (C4), n-pentyl (C5), 3-pentanyl (C5), neopentyl (C5), 3-methyl-2-butanyl (C5), tertiary amyl (C5) and n-hexyl (C6). Further examples of the alkyl group include n-heptyl (C7), n-octyl (C8) and the like. Unless otherwise specified, each occurrence of the alkyl group is independently optionally substituted, i.e., unsubstituted ("unsubstituted alkyl") or substituted with one or more substituents ("substituted alkyl"); for example, 1 to 5 substituents, 1 to 3 substituents, or 1 substituent. In some embodiments, the alkyl group is an unsubstituted C1-10 alkyl (e.g., -CH3). In some embodiments, the alkyl group is a substituted C1-10 alkyl.

[0043] “Alkenyl” refers to a straight or branched chain hydrocarbon group having from 2 to 20 carbon atoms, one or more carbon-carbon double bonds (e.g., 1, 2, 3, or 4 carbon-carbon double bonds), and optionally one or more carbon-carbon triple bonds (e.g., 1, 2, 3, or 4 carbon-carbon triple bonds) (“C2-20 alkenyl”). In some embodiments, the alkenyl does not contain any triple bonds. In some embodiments, the alkenyl group contains from 2 to 10 carbon atoms (“C2-10 alkenyl”). In some embodiments, the alkenyl group contains from 2 to 9 carbon atoms (“C2-9 alkenyl”). In some embodiments, the alkenyl group contains from 2 to 8 carbon atoms (“C2-8 alkenyl”). In some embodiments, the alkenyl group comprises 2 to 7 carbon atoms ("C2-7 alkenyl"). In some embodiments, the alkenyl group comprises 2 to 6 carbon atoms ("C2-6 alkenyl"). In some embodiments, the alkenyl group comprises 2 to 5 carbon atoms ("C2-5 alkenyl"). In some embodiments, the alkenyl group comprises 2 to 4 carbon atoms ("C2-4 alkenyl"). In some embodiments, the alkenyl group comprises 2 to 3 carbon atoms ("C2-3 alkenyl"). In some embodiments, the alkenyl group comprises 2 carbon atoms ("C2 alkenyl"). The one or more carbon-carbon double bonds can be internal (such as in 2-butenyl) or terminal (such as in 1-butenyl). Examples of the C2-4 alkenyl group include ethenyl (C2), 1-propenyl (C3), 2-propenyl (C3), 1-butenyl (C4), 2-butenyl (C4), butadienyl (C4) and the like. Examples of the C2-6 alkenyl group include the above-mentioned C2-4 alkenyl groups as well as pentenyl (C5), pentadienyl (C5), hexenyl (C6) and the like. Further examples of alkenyl include heptenyl (C7), octenyl (C8), octatrienyl (C8) and the like. Unless otherwise indicated, each occurrence of an alkenyl group is independently optionally substituted, i.e., unsubstituted ("unsubstituted alkenyl") or substituted with one or more substituents ("substituted alkenyl"), such as 1 to 5 substituents, 1 to 3 substituents, or 1 substituent. In some embodiments, the alkenyl group is an unsubstituted C2-10 alkenyl. In some embodiments, the alkenyl group is a substituted C2-10 alkenyl.

[0044] “Alkynyl” refers to a straight or branched chain hydrocarbon group having from 2 to 20 carbon atoms, one or more carbon-carbon triple bonds (e.g., 1, 2, 3, or 4 carbon-carbon triple bonds), and optionally one or more carbon-carbon double bonds (e.g., 1, 2, 3, or 4 carbon-carbon double bonds) (“C2-20 alkynyl”). In some embodiments, an alkynyl does not contain any double bonds. In some embodiments, an alkynyl group contains from 2 to 10 carbon atoms (“C2-10 alkynyl”). In some embodiments, an alkynyl group contains from 2 to 9 carbon atoms (“C2-9 alkynyl”). In some embodiments, an alkynyl group contains from 2 to 8 carbon atoms (“C2-8 alkynyl”). In some embodiments, the alkynyl group comprises 2 to 7 carbon atoms ("C2-7 alkynyl"). In some embodiments, the alkynyl group comprises 2 to 6 carbon atoms ("C2-6 alkynyl"). In some embodiments, the alkynyl group comprises 2 to 5 carbon atoms ("C2-5 alkynyl"). In some embodiments, the alkynyl group comprises 2 to 4 carbon atoms ("C2-4 alkynyl"). In some embodiments, the alkynyl group comprises 2 to 3 carbon atoms ("C2-3 alkynyl"). In some embodiments, the alkynyl group comprises 2 carbon atoms ("C2 alkynyl"). The one or more carbon-carbon triple bonds can be internal (such as in 2-butynyl) or terminal (such as in 1-butynyl). Examples of the C2-4 alkynyl group 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 the C2-6 alkenyl group include the aforementioned C2-4 alkynyl group, as well as pentynyl (C5), hexynyl (C6) and the like. Further examples of the alkynyl group include heptynyl (C7), octynyl (C8) and the like. Unless otherwise specified, each occurrence of the alkynyl group is independently optionally substituted, i.e., unsubstituted ("unsubstituted alkynyl") or substituted ("substituted alkynyl") with one or more substituents; for example, 1 to 5 substituents, 1 to 3 substituents, or 1 substituent. In some embodiments, the alkynyl group is an unsubstituted C2-10 alkynyl. In some embodiments, the alkynyl group is a substituted C2-10 alkynyl.

[0045] "Aryl" refers to a monocyclic or polycyclic (e.g., bicyclic or tricyclic) 4n+2 aromatic ring system (e.g., containing 6, 10, or 14 π-electrons shared by the cyclic structure) containing 6-14 ring carbon atoms and zero heteroatoms located within the aromatic ring system ("C6-14 aryl"). In some embodiments, the aryl group contains a ring of six carbon atoms ("C6 aryl"; e.g., phenyl). In some embodiments, the aryl group contains ten ring carbon atoms ("C10 aryl"; e.g., naphthyl, such as 1-naphthyl and 2-naphthyl). In some embodiments, the aryl group contains fourteen ring carbon atoms ("C14 aryl"; e.g., anthracyl). "Aryl" also includes ring systems in which an aryl ring as defined above is fused to one or more carbocyclyl or heterocyclyl groups where the radical or point of attachment is on the aryl ring, and in such cases the number of carbon atoms continues to denote the number of carbon atoms in the aryl ring system. Common aryl groups include, but are not limited to, those derived from aceanthrilene, acenaphthylene, acephenanthrylene, anthracene, azulene, benzene, chrysene, coronene, fluoranthene, fluorene, hexacene, hexaphene, hexalene, ac-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. Specific aryl groups include phenyl, naphthyl, indenyl, and tetrahydronaphthyl. Unless otherwise indicated, each occurrence of an aryl group is independently optionally substituted, i.e., unsubstituted ("unsubstituted aryl") or substituted ("substituted aryl") with one or more substituents. In some embodiments, the aryl group is unsubstituted C6-14 aryl. In some embodiments, the aryl group is substituted C6-14 aryl.

[0046] "Heteroaryl" refers to a radical of a 5- to 10-membered monocyclic or bicyclic 4n+2 aromatic ring system (e.g., containing 6 or 10 π-electrons common to the cyclic structure) containing ring carbon atoms and 1-4 ring heteroatoms present in the aromatic ring system, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur ("5- to 10-membered heteroaryl"). In heteroaryl groups that contain one or more nitrogen atoms, the point of attachment may be a carbon or nitrogen atom, if valence permits. Heteroaryl bicyclic ring systems may contain one or more heteroatoms in one or both rings. "Heteroaryl" includes ring systems in which a heteroaryl ring as defined above is fused to one or more carbocyclyl or heterocyclyl groups, where the point of attachment is on the heteroaryl ring, in which cases the number of ring members continues to denote 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 to one or more aryl groups, where the point of attachment is on either the aryl or heteroaryl ring, in which cases the number of ring members denotes the number of ring members in the fused (aryl/heteroaryl) ring system. In bicyclic heteroaryl groups in which one ring does not contain a heteroatom (e.g., indolyl, quinolinyl, carbazolyl, and the like groups), the point of attachment can be on either ring, i.e., either on the ring bearing the heteroatom (e.g., 2-indolyl) or on the ring that does not contain the heteroatom (e.g., 5-indolyl). In some embodiments, the heteroaryl group is a 5- to 10-membered aromatic ring system containing ring carbon atoms and 1-4 ring heteroatoms located in the aromatic ring system, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur ("5- to 10-membered heteroaryl"). In some embodiments, the heteroaryl group is a 5- to 8-membered aromatic ring system comprising carbon ring atoms and 1-4 ring heteroatoms located within the aromatic ring system, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur ("5- to 8-membered heteroaryl"). In some embodiments, the heteroaryl group is a 5- to 6-membered aromatic ring system comprising carbon ring atoms and 1-4 ring heteroatoms located within the aromatic ring system, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur ("5- to 6-membered heteroaryl"). In some embodiments, the 5- to 6-membered heteroaryl comprises 1-3 ring heteroatoms selected from nitrogen, oxygen, and sulfur. In some embodiments, the 5- to 6-membered heteroaryl comprises 1-2 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 noted, each occurrence of the heteroaryl group is independently optionally substituted, i.e., unsubstituted ("unsubstituted heteroaryl") or substituted ("substituted heteroaryl") with one or more substituents. In some embodiments, the heteroaryl group is an unsubstituted 5- to 14-membered heteroaryl. In some embodiments, the heteroaryl group is substituted with a 5- to 14-membered heteroaryl. Representative 5-membered heteroaryl groups containing one heteroatom include, but are not limited to, pyrrolyl, furanyl, and thiophenyl. Representative 5-membered heteroaryl groups containing two heteroatoms include, but are not limited to, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, and isothiazolyl. Representative 5-membered heteroaryl groups containing three heteroatoms include, but are not limited to, triazolyl, oxadiazolyl, and thiadiazolyl. Representative 5-membered heteroaryl groups containing four heteroatoms include, but are not limited to, tetrazolyl. Representative 6-membered heteroaryl groups containing one heteroatom include, but are not limited to, pyridinyl. Representative 6-membered heteroaryl groups containing two heteroatoms include, but are not limited to, pyridazinyl, pyrimidinyl, and pyrazinyl. Representative 6-membered heteroaryl groups containing three or four heteroatoms include, but are not limited to, triazinyl and tetrazinyl, respectively. Representative 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, benzoisofuranyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl, benzoxadiazolyl, benzthiazolyl, benzisothiazolyl, benzthiadiazolyl, indolzinyl, and purinyl. Examples of 6,6-bicyclic heteroaryl groups include, but are not limited to, naphthyridinyl, pteridinyl, quinolinyl, isoquinolinyl, cinnolinyl, quinoxalinyl, phthalazinyl, and quinazolinyl.

[0047] "Carbocyclyl" or "carbocyclic" refers to a radical of a non-aromatic cyclic hydrocarbon group comprising 3 to 10 ring carbon atoms ("C3-10 carbocyclyl") and zero heteroatoms in the non-aromatic ring system. In some embodiments, the carbocyclyl group comprises 3 to 8 ring carbon atoms ("C3-8 carbocyclyl"). In some embodiments, the carbocyclyl group comprises 3 to 6 ring carbon atoms ("C3-6 carbocyclyl"). In some embodiments, the carbocyclyl group comprises 3 to 6 ring carbon atoms ("C3-6 carbocyclyl"). In some embodiments, the carbocyclyl group comprises 5 to 10 ring carbon atoms ("C5-10 carbocyclyl"). Typical 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), and the like. Typical C3-8 carbocyclyl groups include, but are not limited to, the above C3-6 carbocyclyl group and cycloheptyl (C7), cycloheptenyl (C7), cycloheptadienyl (C7), cycloheptatrienyl (C7), cyclooctyl (C8), cyclooctenyl (C8), bicyclo[2,2,1]heptyl (C7), bicyclo[2,2,2]octyl (C8), and the like. Typical C3-10 carbocyclyl groups include, but are not limited to, the above-mentioned C3-8 carbocyclyl group and cyclononyl (C9), cyclononenyl (C9), cyclodecyl (C10), cyclodecenyl (C10), octahydro-1H-indenyl (C9), decahydronaphthyl (C10), spiro[4,5]decyl (C10), etc. The preceding examples illustrate that in some embodiments, the carbocyclyl group is a monocyclic ring ("monocyclic carbocyclyl") or a carbocyclyl group that contains a fused ring system, a bridged ring system, or a spiro ring system, such as a bicyclic ring system ("bicyclic carbocyclyl"), and may be a saturated or partially unsaturated carbocyclyl group. "Carbocyclyl" also includes ring systems in which the aforementioned carbocyclic ring is fused to one or more aryl or heteroaryl groups, wherein the point of attachment is on the carbocyclic ring, in which cases the number of carbon atoms continues to be the number of carbon atoms in the carbocyclic ring system. Unless otherwise noted, each of the carbocyclyl groups is independently optionally substituted, i.e., unsubstituted (an "unsubstituted carbocyclyl") or substituted with one or more substituents (a "substituted carbocyclyl"). In some embodiments, the carbocyclyl is an unsubstituted C3-10 carbocyclyl. In some embodiments, the carbocyclyl is a substituted C3-10 carbocyclyl. In some embodiments, the "carbocyclyl" is a monocyclic saturated carbocyclyl ("C3-10 cycloalkyl") having from 3 to 10 ring carbon atoms. 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 ("C3-6 cycloalkyl"). In some embodiments, a cycloalkyl group has 5 to 6 ring carbon atoms ("C5-6 cycloalkyl"). In some embodiments, cycloalkyl groups have 5 to 10 ring carbon atoms ("C5-10 cycloalkyl"). Examples of C5-6 cycloalkyl include cyclopentyl (C5) and cyclohexyl (C6). Examples of a C3-6 cycloalkyl group include the above C5-6 cycloalkyl groups, as well as cyclopropyl (C3) and cyclobutyl (C4). Examples of a C3-8 cycloalkyl group include the above C3-6 cycloalkyl group, as well as cycloheptyl (C7) and cyclooctyl (C8). Unless otherwise indicated, each instance of a cycloalkyl group is independently unsubstituted ("unsubstituted cycloalkyl") or substituted ("substituted cycloalkyl") with one or more substituents. In some embodiments, cycloalkyl is unsubstituted C3-10 cycloalkyl. In some embodiments, cycloalkyl is substituted C3-10 cycloalkyl.

[0048] "Heterocyclyl" or "heterocyclic" refers to a radical of a 3- to 10-membered non-aromatic ring system containing ring carbon atoms and from 1 to 4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, sulfur, boron, phosphorus, and silicon ("3- to 10-membered heterocyclyl"). In heterocyclic groups containing one or more nitrogen atoms, the point of attachment may be a carbon or nitrogen atom, if valence permits. A heterocyclyl group may be either a monocyclic ("monocyclic heterocyclyl") or a fused, bridged, or spiro ring system, such as a bicyclic system ("bicyclic heterocyclyl"), and may be saturated or may be partially unsaturated. Heterocyclyl bicyclic ring systems may 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, wherein the point of attachment is on either the carbocyclyl or heterocyclyl ring, or ring systems in which a heterocyclyl ring, as defined above, is fused to one or more aryl or heteroaryl groups, wherein the point of attachment is on the heterocyclyl ring, in which cases the number of ring members continues to denote the number of ring members in the heterocyclyl ring system. Unless otherwise indicated, each instance of heterocyclyl is independently optionally substituted, i.e., unsubstituted ("unsubstituted heterocyclyl") or substituted ("substituted heterocyclyl") with one or more substituents. In some embodiments, the heterocyclyl group is an unsubstituted 3- to 10-membered heterocyclyl. In some embodiments, the heterocyclyl group is a substituted 3- to 10-membered heterocyclyl. In some embodiments, the heterocyclyl group is a 5- to 10-membered non-aromatic ring system comprising carbon ring atoms and 1-4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, sulfur, boron, phosphorus, and silicon ("5- to 10-membered heterocyclyl"). In some embodiments, the heterocyclyl group is a 5- to 8-membered non-aromatic ring system comprising carbon ring atoms and 1-4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur ("5- to 8-membered heterocyclyl"). In some embodiments, the heterocyclyl group is a 5- to 6-membered non-aromatic ring system comprising carbon ring atoms and 1-4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur ("5- to 6-membered heterocyclyl"). In some embodiments, the 5- to 6-membered heterocyclyl contains 1-3 ring heteroatoms selected from nitrogen, oxygen, and sulfur. In some embodiments, the 5- to 6-membered heterocyclyl contains 1-2 ring heteroatoms selected from nitrogen, oxygen, and sulfur. In some embodiments, the 5- to 6-membered heterocyclyl contains one ring heteroatom selected from nitrogen, oxygen, and sulfur.

[0049] "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 have been replaced by a nitrogen, oxygen or sulfur heteroatom. Hetero can be used with respect to any of the hydrocarbyls described above: for example, alkyl, for example heteroalkyl; cycloalkyl groups, for example heterocyclyl groups; aryl, for example heteroaryl; cycloalkenyl groups, for example cycloheteroalkenyl and the like; containing from 1 to 5 heteroatoms, in particular from 1 to 3 heteroatoms.

[0050] "Alkoxy" refers to the group -OR, wherein 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., with 1 to 6 carbon atoms. Additional specific alkoxy groups contain 1 to 4 carbon atoms. Typical substituted alkoxy groups include, but are not limited to, -O-( _CH2 ) _t ( _C6 - _C10 aryl), -O-( _CH2 ) _t (5-10 membered heteroaryl), -O-( _CH2 ) _t ( _C3 - _C10 cycloalkyl), and -O-( _CH2 ) _t (4-10 membered heterocyclyl), where t is an integer from 0 to 4 and any substituted groups present, including aryl, heteroaryl, cycloalkyl, or heterocyclyl groups, may themselves be substituted with an unsubstituted _C1 - _C4 alkyl group, a halogen, an unsubstituted _C1 - _C4 alkoxy group, an unsubstituted _C1 - _C4 haloalkyl group, an unsubstituted _C1 -C ₄ hydroxyalkyl group or an unsubstituted C ₁ -C ₄ haloalkoxy group or hydroxy group.

[0051] "Carboxy" refers to -C(O)OH.

[0052] "Cyano" refers to -CN.

[0053] "Halo" or "halogen" refers to a fluorine atom (F), a chlorine atom (Cl), a bromine atom (Br), and an iodine atom (I). In some embodiments, the halogen group is either a fluorine atom or a chlorine atom.

[0054] "Hydroxy" refers to -OH.

[0055] "Nitro" refers to -NO ₂ .

[0056] Other definitions

[0057] The term "pharmaceutically acceptable salt" refers to those salts which are medically acceptable for use in contact with the tissues of humans and lower animals without causing undue toxicity, irritation, allergic response, and the like, in accordance with a reasonable benefit/risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, Berge et al. describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences (1977) 66:1-19. Pharmaceutically acceptable salts of the compounds of the present invention include salts derived from suitable inorganic and organic acids and bases. Examples of pharmaceutically acceptable, non-toxic acid addition salts are salts of the amino group 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 obtained by 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, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxyethanesulfonate, lactobionate, lactate, laurate, lauryl 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 salts and the like. Pharmaceutically acceptable salts derived from suitable bases include alkali metal, alkaline earth metal, ammonium and N+(C1-4alkyl)4 salts. Examples of alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium and the like. Additional pharmaceutically acceptable salts include, where appropriate, non-toxic ammonium, quaternary ammonium and amine cations formed using counterions such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, lower alkyl sulfonate and aryl sulfonate.

BRIEF DESCRIPTION OF DRAWINGS

[0058] Figure 1 is a diagram showing changes in plasma and brain control over time.

[0059] Figure 2 is a graph showing changes in KH022 in plasma and brain over time.

DETAILED DESCRIPTION OF CERTAIN EMBODIMENTS OF THE PRESENT INVENTION

[0060] In order to better explain the technical scheme of the present invention, the present invention provides synthesis examples or biological examples of a part of the compounds, and they do not further limit the scope of legal protection of the present invention.

[0061] Materials and methods

[0062] The compounds disclosed herein can be prepared from readily available starting materials using the following general methods and procedures. It should be apparent that under the usual or preferred process conditions (i.e., reaction temperature, reaction time, molar ratio of reactants, solvent, pressure, etc.), other process conditions may also be used unless otherwise noted. Optimum reaction conditions may vary depending on the particular reactants or solvents used, but such conditions can be determined by those skilled in the art through routine optimization.

[0063] In addition, it will be apparent to those skilled in the art that conventional protecting groups may be required to prevent undesirable reactions of certain functional groups. Methods for selecting suitable protecting groups for a particular functional group and selecting suitable conditions for protecting and deprotecting the groups are well known in the art.

[0064] The compounds provided herein can be isolated and purified using known standard techniques, including but not limited to recrystallization, column chromatography, HPLC, or supercritical fluid chromatography (SFC). Details for the preparation of representative pyrazoles are provided herein in the following reaction schemes.

[0065] The compounds provided herein can be prepared according to methods described in the art using suitable reagents, starting materials and purification methods known to those skilled in the art.

[0066] Example 1: Synthesis of compound KH001:

THF

THF
[0067]

[0068] KHC-2 compound: KCN (600.0 g, 9.2 mol), acetic acid (640 mL) was added to a solution of KHC-1 compound (100.0 g, 0.3 mol) in methanol (4 L), and the reaction mixture was stirred overnight at room temperature (RT). The reaction solution was poured into water and extracted with ethyl acetate (EA). The organic phase was dried over anhydrous sodium sulfate and concentrated under vacuum to give a crude product, which was dissolved in acetic acid (2.5 L). Platinum dioxide (24.0 g, 0.1 mol) was added to the reaction solution, and hydrogenation was performed with H ₂ gas. Thin layer chromatography (TLC) showed the reaction to be complete and the reaction solution was filtered, then the filtered solution was concentrated to approximately 200 mL, diluted with ice-cold water (2.4 L). Aqueous _NaNO2 (46.0 g, 0.7 mol) was slowly added dropwise to the mixture, then the mixture was stirred for 15 h at RT and extracted with EA. The combined organic layers were washed with brine, dried, concentrated and purified by flash column chromatography on silica gel to give KHC-2 (57.3 g, 55% yield).

[0069] KHC-3 Compound: Ethynyl magnesium bromide (395 mL, 0.5 M) was added dropwise to a solution of KHC-2 (57.3 g, 164.5 mmol) in THF (1 L) under N ₂ atmosphere at -78 °C, then gradually warmed to RT, stirred overnight at RT. Saturated aqueous NH ₄ Cl solution was slowly added to the above mixture, followed by extraction with EA, washing the organic phase with brine, drying over anhydrous Na ₂ SO ₄ , concentrating and purifying the resulting crude product by flash chromatography on a silica gel column to give KHC-3 Compound (33.7 g, yield 55%).

[0070] 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), followed by refluxing with the addition of Dowex-50 (10.0 g). After completion of the reaction, cooling to RT, filtering the resin, washing with EA, neutralizing the filtrate with 40% aqueous NaOH, washing the organic layer with saturated brine, drying over anhydrous Na ₂ SO ₄ , concentrating and purifying the resulting crude product by flash chromatography on a silica gel column to give KHC-4 Compound (23.6 g, yield 70%)

[0071] KHC-5 Compound: KHC-4 (23.6 g, 63.3 mmol) and palladium black (1.2 g) in THF (200 mL) were hydrogenated with H ₂ gas, after stirring overnight at RT, the mixture was filtered through a celite pad, washed with EA and the filtrate was concentrated in vacuo to give the crude compound, which was recrystallized from acetone to give KHC-5 (19.0 g, 80% yield).

[0072] 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 solution was adjusted with dilute HCl, and then the solution was extracted with EA. The organic phase was washed with brine, dried over anhydrous Na ₂ SO ₄ and concentrated to give the crude product, which was purified by flash column chromatography on silica gel to give compound KH001 (13.5 g, yield 80%)

^1H NMR (400 MHz, CDCl ₃ ), 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 ).

[0073] Example 2: Synthesis of compound KH002:

[0074]

[0075] Compound KHC-6: Compound KH001 (1.5 g, 4.5 mmol) synthesized in Example 1 was dissolved in _CH3OH (40 mL) until the solution became clear, then the solution was cooled to 5°C on water ice. Then, bromine (1.5 g, 9.2 mmol) was added dropwise to the cooled solution. The ice bath was removed and the reaction was carried out at RT for 4 hours, the reaction solution turned from orange to light yellow. Monitoring the reaction progress by TLC showed that the reaction was complete, water (30 ml) was added to the reaction solution, extraction was performed with EA (2x35 ml), the combined organic layers were washed with saturated brine, dried over _{anhydrous Na2SO4} and the solvent was distilled off under reduced pressure to obtain a crude light yellow solid, which was purified by flash column chromatography on silica gel (eluent: _CH2Cl2 ) to obtain a white solid (1.0 g, yield _56.7 %).

[0076] Compound KH002: Compound KHC-6 (80 mg, 0.2 mmol) was dissolved in THF (5 mL) until the solution became clear, then K ₂ CO ₃ (83 mg, 0.6 mmol) and 4-cyanopyrazole (56 mg, 0.6 mmol) were added. The reaction was carried out at room temperature overnight. Monitoring the reaction progress by TLC showed that the reaction was complete, then the reaction mixture was washed with 10 ml of water and extracted with EA (2x20 ml), the combined organic layers were washed with saturated brine, dried over _{anhydrous Na2SO4} and the solvent was distilled off under reduced pressure to obtain a crude light yellow solid, which was purified by flash chromatography on a silica gel column ₍ eluent: _CH2Cl2 / _CH3OH =100:1), and then recrystallized (EA/petroleum ether (PE)=1:10) to obtain a white solid (36 mg, yield 42.5%). ^1H NMR (400 MHz, CDCl ₃ ) 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).

[0077] Example 3: Synthesis of compound KH003:

[0078]

[0079] Compound KH003: Compound KHC-6 (50 mg, 0.1 mmol) was dissolved in THF (5 mL) until the solution became clear, then K ₂ CO ₃ (50 mg, 0.3 mmol) and 1H-triazole (25 mg, 0.3 mmol) were added. The reaction was carried out at room temperature overnight. Monitoring the reaction progress by TLC showed that the reaction was complete, then the reaction mixture was washed with 10 ml of water, extracted with EA (2x20 ml), the combined organic layers were washed with saturated brine, dried over _{anhydrous Na2SO4} and the solvent was distilled off under reduced pressure to obtain a crude light yellow solid, which was purified by flash chromatography on a silica gel column ₍ eluent: _CH2Cl2 / _CH3OH =150:1), and then recrystallized (EA/PE=1:10) to obtain a white solid (17 mg, yield 35.5%). ^1H NMR (400 MHz, CDCl ₃ ) 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).

[0080] Example 4: Synthesis of compound KH004 and KH005:

CT
[0081]

[0082] Compound KH004 and KH005: Compound KHC-6 (103 mg, 0.2 mmol) was dissolved in CH ₃ CN (15 mL) until the solution became clear, then Cs ₂ CO ₃ (244 mg, 0.7 mmol) and 1H-tetrazole (53 mg, 0.7 mmol) were added. The reaction was carried out at room temperature. Monitoring the reaction progress by TLC 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 layers were washed with saturated brine, dried over _{anhydrous Na2SO4} and the solvent was distilled off under reduced pressure to give a crude light yellow solid, which was purified by flash column chromatography on silica gel (eluent: _CH2Cl2 / _CH3OH = from 150:1 to 90:1) to give two compounds _, which were respectively recrystallized (EA/PE=1:10) to give white solid one (24 mg, 24.0%) and white solid two (36 mg, yield 36.0%).

White solid 1 (KH004): ^1H NMR (400 MHz, _CDCl3 ) δ 8.57 (s, 1H), 5.50 (s, 2H), 4.06-4.05 (m, 1H), 2.35 (dd, 1H), 1.92-1.81 (m, 3H), 1.74-1.68 (m, 3H), 1.64-1.57 (m, 4H), 1.46-1.11 (m, 10H), 1.05-0.98 (m, 2H), 0.96 (s, 3H), 0.88-0.78 (m, 3H), 0.75 (s, 3H).

White solid 2 (KH005): ^1H NMR (400 MHz, _CDCl3 ) δ 8.71 (s, 1H), 5.38-5.23 (m, 2H), 4.06-4.05 (m, 1H), 2.41 (dd, 1H), 2.28 (dd, 1H), 1.91-1.72 (m, 4H), 1.68-1.51 (m, 7H), 1.47-1.11 (m, 10H), 1.06-0.99 (m, 1H), 0.95 (s, 3H), 0.91-0.78 (m, 3H), 0.75 (s, 3H).

[0083] Example 5: Synthesis of compound KH006:

CT
[0084]

[0085] Compound KH006: Compound KHC-6 (50 mg, 0.1 mmol) was dissolved in CH ₃ CN (8 mL), then Cs ₂ CO ₃ (117 mg, 0.3 mmol) and 1H-pyrazole (24 mg, 0.3 mmol) were added. The reaction was carried out at room temperature overnight. Monitoring the reaction progress by TLC 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 layers were washed with _saturated brine, dried over _{anhydrous Na2SO4} and the solvent was distilled off under reduced pressure to obtain a crude light yellow solid, which was purified by flash chromatography on a silica gel column (eluent: _CH2Cl2 / _CH3OH = 100:1) and recrystallized (EA/PE = 1:10) to obtain a white solid (24 mg, yield 50.2%). ^1H NMR (400 MHz, CDCl ₃ ) 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).

[0086] Example 6: Synthesis of compound KH007:

[0087]

[0088] Compound KH007: Compound KHC-6 (150 mg, 0.3 mmol) was dissolved in THF (10 mL), then K ₂ CO ₃ (152 mg, 1.1 mmol) and 1H-imidazole (75 mg, 1.1 mmol) were added. The reaction was carried out at room temperature overnight. Monitoring the reaction progress by TLC showed that the reaction was complete. Water (10 ml) was added to the reaction solution, extraction was performed with EA (2x20 ml), the combined organic layers _were washed with saturated brine, dried over _{anhydrous Na2SO4} and the solvent was distilled off under reduced pressure to obtain a crude light yellow solid, which was purified by flash chromatography on a silica gel column (eluent: _CH2Cl2 / _CH3OH =30:1) followed by recrystallization (EA) to obtain a white solid (35 mg, yield 24.1%). ^1H NMR (400 MHz, CDCl ₃ ) 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), 1.56 (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).

[0089] Example 7: Synthesis of compound KH008:

[0090]

[0091] Compound KH008: Compound KHC-6 (100 mg, 0.2 mmol) was dissolved in THF (10 mL), then K ₂ CO ₃ (97 mg, 0.7 mmol) and 3-cyanopyrazole (68 mg, 0.7 mmol) were added. The reaction was carried out at 70°C for 2 hours. Monitoring the reaction progress by TLC showed that the reaction was complete, water (10 ml) was added to the reaction solution, extraction was performed with EA (2x20 ml), the combined organic layers were washed with saturated brine, dried over _{anhydrous Na2SO4} and the solvent was distilled off under reduced pressure to obtain a crude light yellow solid, which was purified by TLC (eluent: PE/acetone = 3:1) followed by recrystallization ( _CH3CN ) to obtain a white solid (20 mg, yield 19.4%). ^1H NMR (400 MHz, CDCl ₃ ) δ 7.44 (d, 1H), 6.72 (d, 1H), 6.34 (s, 1H), 5.00 (k, 2H), 4.06-4.05 (m, 1H), 2.32 (dd, 1H), 1.88-1.68 (m, 4H), -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).

[0092] Example 8: Synthesis of compound KH009:

CT
[0093]

[0094] Compound KH009: Compound KHC-6 (50 mg, 0.1 mmol) was dissolved in CH ₃ CN (5 mL) until clear, then Cs ₂ CO ₃ (120 mg, 0.3 mmol) and 1H-indole (68 mg, 0.7 mmol) were added. The reaction was carried out at room temperature overnight. Monitoring the reaction progress by TLC showed that the reaction was complete. Water (10 ml) was added to the reaction solution, extraction was performed with EA (2×20 ml), the combined organic layers were washed with saturated brine, dried over _{anhydrous Na2SO4 ,} and the solvent was distilled off under reduced pressure to obtain a crude light yellow solid, which was purified by TLC (eluent: _CH2Cl2 /EA=10:1) to obtain a light yellow solid (34 mg, yield 62.4%). ^1H NMR (400 MHz, CDCl ₃ ) 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), 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).

[0095] Example 9: Synthesis of compound KH010:

[0096]

[0097] Compound KH010: Compound KHC-6 (100 mg, 0.2 mmol) was dissolved in CH ₃ CN (5 mL), then K ₂ CO ₃ (101 mg, 0.7 mmol) and 4-(trifluoromethyl)-1H-pyrazole (35 mg, 0.2 mmol) were added. The reaction was carried out at 50 °C for 2 h. Monitoring the reaction progress by TLC showed that the reaction was complete. Water (10 ml) was added to the reaction solution, extraction was performed with EA (2x20 ml), the combined organic layers were washed with saturated brine, dried over _{anhydrous Na2SO4} and the solvent was distilled off under reduced pressure to obtain a crude light yellow solid, which was purified by flash chromatography on a silica gel column (eluent: PE/acetone = 7:1) followed by recrystallization ( _CH3CN ) to obtain a white solid (61 mg, yield 53.1%). ^1H NMR (400 MHz, CDCl ₃ ) δ 7.71 (d, 2H), 4.98 (k, 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).

[0098] Example 10: Synthesis of Compound KH011:

CT
[0099]

[0100] Compound KH011: Compound KHC-6 (50 mg, 0.1 mmol) was dissolved in CH ₃ CN (5 mL), then Cs ₂ CO ₃ (120 mg, 0.3 mmol) and 1H-indazole (43 mg, 0.3 mmol) were added. The reaction was carried out at room temperature overnight. Monitoring the reaction progress by TLC showed that the reaction was complete. Water (10 ml) was added to the reaction solution, extraction was performed with EA (2×20 ml), the combined organic layers were washed with saturated brine, dried over _{anhydrous Na2SO4} and the solvent was distilled off under reduced pressure to obtain a crude light yellow solid, which was purified by TLC (eluent: PE/acetone = 3:1) followed by recrystallization ( _CH3CN ) to obtain a white solid (20 mg, 36.6%). ^1H NMR (400 MHz, CDCl ₃ ) 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).

[0101] Example 11: Synthesis of compound KH012:

[0102]

[0103] Compound KH012: Compound KHC-6 (50 mg, 0.1 mmol) was dissolved in CH ₃ CN (5 mL), then Cs ₂ CO ₃ (120 mg, 0.3 mmol) and 1H-purine (44 mg, 0.3 mmol) were added. The reaction was carried out at room temperature overnight. Monitoring the reaction progress by TLC showed that the reaction was complete. Water (10 ml) was added to the reaction solution, the mixture was extracted with EA (2x20 ml), the combined organic layers were washed with saturated brine _, dried over _{anhydrous Na2SO4} and the solvent was distilled off under reduced pressure to obtain the crude product as a light yellow solid, which was purified by TLC (eluent: _CH2Cl2 / _CH3OH =20:1), followed by recrystallization ( _CH3CN ) to obtain a white solid (5 mg, yield 9.1%). ^1H NMR (400 MHz, CDCl ₃ ) δ 9.22 (s, 1H), 9.06 (s, 1H), 8.25 (s, 1H), 5.18 (k, 2H), 4.07-4.06 (m, 1H), 2.49 (dd, 1H), 1.93-1.72 (m, 4H), -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).

[0104] Example 12: Synthesis of compound KH013:

[0105]

[0106] Compound KH013: Compound KHC-6 (80 mg, 0.2 mmol) was dissolved in CH ₃ CN (8 mL), then Cs ₂ CO ₃ (196 mg, 0.6 mmol) and 4-chloropyrazole (62 mg, 0.6 mmol) were added. The reaction was carried out at room temperature overnight. Monitoring the reaction progress by TLC showed that the reaction was complete. 15 ml of water were added to the reaction mixture, the mixture was extracted with EA (2x30 ml), the combined organic layers were washed with saturated brine, dried over _{anhydrous Na2SO4} and the solvent _was distilled off under reduced pressure to obtain a crude light yellow solid, which was purified by flash chromatography on a silica gel column (eluent: _CH2Cl2 / _CH3OH =100:1) and then recrystallized (EA/PE=1:10) to obtain a white solid (28 mg, yield 32.3%). ^1H NMR (400 MHz, CDCl ₃ ) 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).

[0107] Example 13: Synthesis of compound KH014:

[0108]

[0109] Compound KH014: Compound KHC-6 (60 mg, 0.15 mmol) was dissolved in DMF (5 mL), then NaOH (1 mL, 1.0 mmol) was added. The reaction was carried out at room temperature overnight. Monitoring the reaction progress by TLC showed that the reaction was complete. 10 mL of water was added to the reaction mixture, the mixture was extracted with EA (2 × 20 mL), the combined organic layers were washed with saturated NaCl, dried over anhydrous Na ₂ SO ₄ and the solvent was distilled off under reduced pressure to obtain a crude product, which was purified by TLC (eluent: CH ₂ Cl ₂ /CH ₃ OH = 50:1) to give a white solid (12 mg, yield 22.92%). ^1H 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).

[0111] Example 14: Synthesis of compound KH015:

[0112]

[0113] Compound KH015: Compound KHC-6 (60 mg, 0.15 mmol) was dissolved in CH ₃ CN (5 mL), then Cs ₂ CO ₃ (98 mg, 0.30 mmol) and 6-chloropurine (46 mg, 0.30 mmol) were added. The reaction was carried out at room temperature overnight. Monitoring the reaction progress by TLC showed that the reaction was complete. 10 ml of water were added to the reaction mixture, the mixture was extracted with EA (2 × 20 ml), the combined organic layers were washed with saturated NaCl, dried over anhydrous Na ₂ SO ₄ and the solvent was distilled off under reduced pressure to obtain a crude product, which was purified by TLC (eluent: CH ₂ Cl ₂ /CH ₃ OH = 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 (k, 2H), 4.20 (d, 1H), 3 .79 (t, 1H), 2.56 (dd, 1H), 1.80 - 0.75 (m, 27H), 0.69 (s, 3H).

[0115] Example 15: Synthesis of compound KH016:

[0116]

[0117] Compound KH016: KHC-6 (60 mg, 0.15 mmol) was dissolved in 5 mL of CH ₃ CN, then Cs ₂ CO ₃ (98 mg, 0.30 mmol) and 2-mercaptothiophene (17 mg, 0.30 mmol) were added. The reaction was carried out at room temperature overnight. Monitoring the reaction progress by TLC showed that the reaction was complete. 10 ml of water were added to the reaction mixture, the mixture was extracted with EA (2 × 20 ml), the combined organic layers were washed with saturated NaCl, dried over anhydrous Na ₂ SO ₄ and the solvent was distilled off under reduced pressure to obtain a crude product, which was purified by TLC (eluent: CH ₂ Cl ₂ /CH ₃ OH = 40:1) to obtain a white solid (19 mg, yield 28.34%).

^1H NMR (400 MHz, _CDCl3 ) δ 7.35 (d, 1H), 7.14 (d, 1H), 6.95 (dd, 1H), 4.04 (t, 1H), 3.62 (k, 2H), 2.45 (dd, 1H), 1.65 - 0.81 (m, 27H), 0.73 (s, 3H).

[0119] Example 16: Synthesis of compound KH017:

[0120]

[0121] Compound KH017: KHC-6 (60 mg, 0.15 mmol) was dissolved in 5 mL of CH ₃ CN, then Cs ₂ CO ₃ (98 mg, 0.30 mmol) and 6-methoxypurine (45 mg, 0.30 mmol) were added. The reaction was carried out at room temperature overnight. Monitoring the reaction progress by TLC showed that the reaction was complete. 10 ml of water were added to the reaction mixture, the mixture was extracted with EA (2 × 20 ml), the combined organic layers were washed with saturated NaCl, dried over anhydrous Na ₂ SO ₄ and the solvent was distilled off under reduced pressure to obtain a crude product, which was purified by TLC (eluent: CH ₂ Cl ₂ /CH ₃ OH = 40:1) to obtain a white solid (46 mg, yield 63.76%).

^1H NMR (400 MHz, DMSO -d6 ) δ 8.57 (s, 1H), 8.32 (s, 1H), 5.37 (k, 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).

[0123] Example 17: Synthesis of Compounds KH018 and KH019:

[0124]
CT

[0125] Compounds KH018 and KH019: KHC-6 (60 mg, 0.15 mmol) was dissolved in 5 mL of CH ₃ CN, then Cs ₂ CO ₃ (98 mg, 0.30 mmol) and 5-methyl-1H-tetrazole (25 mg, 0.30 mmol) were added. The reaction was carried out at room temperature overnight. Monitoring the reaction progress by TLC showed that the reaction was complete. To the reaction mixture was added 10 ml of water, the mixture was extracted with EA (2 × 20 ml), the combined organic layers were washed with saturated NaCl, dried over _{anhydrous Na2SO4} and the solvent was distilled off under reduced pressure to obtain a crude product, which _was purified by TLC (eluent: _CH2Cl2 / _CH3OH = 40:1) to give a white solid KH6070053-1 (13 mg, 20.88%) and KH6070053-2 (27 mg, 43.37%) as a white solid, respectively.

^1H NMR (400 MHz, CDCl ₃ ) 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).

^1H NMR (400 MHz, CDCl ₃ ) 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).

[0128] Example 18: Synthesis of compound KH020:

[0129]

[0130] Compound KH020: KHC-6 (60 mg, 0.15 mmol) was dissolved in 5 mL of CH ₃ CN, then Cs ₂ CO ₃ (98 mg, 0.30 mmol) and 2-oxa-6-aza-spiro[3,3]heptane (30 mg, 0.30 mmol) were added. The reaction was carried out at room temperature overnight. Monitoring the reaction progress by TLC showed that the reaction was complete. 10 ml of water were added to the reaction mixture, the mixture was extracted with EA (2 × 20 ml), the combined organic layers were washed with saturated NaCl, dried over anhydrous Na ₂ SO ₄ and the solvent was distilled off under reduced pressure to obtain a crude product, which was purified by TLC (eluent: CH ₂ Cl ₂ /CH ₃ OH = 40:1) to obtain a white solid (21 mg, yield 32.56%).

¹ H NMR (400 MHz, CDCl ₃ ) 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).

[0132] Example 19: Synthesis of compound KH021:

THF
[0133]

[0134] KHC-7:KH001 (0.70 g, 2.11 mmol) was dissolved in 100 mL _{of CH2Cl2} and DMP (1.78 g, 4.20 mmol) was added batchwise. The reaction was carried out at room temperature overnight under _N2 . TLC monitoring of the reaction progress showed that the reaction was complete. 40 ml of water were added to the reaction mixture, the mixture was extracted with EA (2x80 ml), the combined organic layers were washed with saturated NaCl, dried over _{anhydrous Na2SO4} and the solvent was distilled off under reduced pressure to obtain a crude product, which was purified by flash chromatography on silica gel (eluent: PE/acetone = 40:1 → 30:1) to obtain KHC-7 as a white solid (0.62 g, yield 89.47%).

[0135] 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-necked flask under _N2 atmosphere, then anhydrous THF (40 mL) was added and the mixture was stirred at RT for 5 min and cooled to -45°. Then _CH3MgBr (15.20 mL, 15.20 mmol) was added dropwise and the mixture was stirred at -40° for 15 min, then compound KHC-7 (0.62 g, 1.88 mmol) dissolved in anhydrous THF was added dropwise. The reaction mixture was warmed to about -20° and stirred for 2 h. Monitoring the reaction progress by TLC showed that 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 layers were washed with saturated NaCl, dried over _{anhydrous Na2SO4} , and the solvent was distilled off under reduced pressure to obtain a crude product, which was purified by silica gel flash chromatography (eluent: PE/acetone=40:1→35:1) to give a white solid KH021 (0.52 g, yield 78.50%).

^1H NMR (400 MHz, CDCl ₃ ) 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).

[0137] Example 20: Synthesis of Compounds KH022 and KH023:

CT
[0138]

[0139]

[0140] Compounds KHC-8 and KHC-9: Compound KH021 (1.25 g, 3.60 mmol) was dissolved in 40 mL of methanol until clear, 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 light yellow, and TLC monitoring showed that the reaction was complete. The reaction mixture was added with 30 ml of water, extracted with EA (2 × 65 ml), the combined organic layers were washed with saturated sodium chloride, dried over anhydrous sodium sulfate and the solvent was distilled off under reduced pressure to obtain a light yellow crude solid, which was purified by flash chromatography on silica gel (eluted with dichloromethane) to obtain a white solid KHC-8 (0.93 g, 60.78%) and a white solid KHC-9 (0.36 g, 19.89%).

[0141] Compounds KH022 and KH023: Compound KHC-8 (100 mg, 0.24 mmol) was dissolved in 5 mL of _acetonitrile until clear, _Cs2CO3 (234 mg, 0.72 mmol) and 1H-tetrazole (50 mg, 0.72 mmol) were added. The reaction was carried out at room temperature overnight and TLC monitoring showed that the reaction was complete. The reaction mixture was added 10 mL of water, extracted with ethyl acetate (2 x 20 mL), the combined organic layers were washed with saturated sodium chloride, dried over anhydrous sodium sulfate and the solvent was distilled off under reduced pressure to obtain the crude product. Thin layer chromatography on a thick silica gel preparative plate was used for purification (eluted with dichloromethane/methanol = 30:1) to give white solid KH022 (20 mg, 20.08%) and white solid KH023 (47 mg, 47.19%), respectively.

^1H NMR (400 MHz, _CDCl3 ) δ 8.56 (s, 1H), 5.54 - 5.45 (m, 2H), 2.35 (dd, 1H), 1.89 - 1.25 ( m, 21H), 1.20 (s, 3H), 0.95 (s, 3H), 0.88 - 0.80 (m, 3H), 0.72 (s, 3H).

^1H NMR (400 MHz, CDCl ₃ ) 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).

[0144] Example 21: Synthesis of compound KH024:

[0145]

[0146] Compound KH024: KHC _- 8 (60 mg, 0.14 mmol) was dissolved in 5 mL of CH3CN, then _Cs2CO3 (91 mg, 0.28 mmol) and 3-cyanopyrazole (26 mg, 0.28 mmol) were added. The reaction was carried out at room temperature overnight. Monitoring by TLC showed that the reaction was complete. 10 mL of water was added to the reaction mixture, the mixture was extracted with EA (2 × 20 mL), the combined organic layers were washed with saturated NaCl, dried over anhydrous _Na2SO4 and the solvent was distilled off under reduced pressure to obtain a crude product, which _was purified by TLC (eluent: _CH2Cl2 / _CH3OH = 40:1) to obtain a white solid KH024 (16 mg, 26.09%)

^1H NMR (400 MHz, CDCl ₃ ) 7.45 (d, 1H), 6.73 (d, 1H), 5.06 (k, 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).

[0148] Example 22: Synthesis of compound KH025:

[0149]

[0150] Compound KH025: KHC-8 (100 mg, 0.24 mmol) was dissolved in 5 ml of DMSO, then _Cs2CO3 ( ₂₃₅ mg, 0.72 mmol) and 3-hydroxybenzoisoxazole (97 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 were added to the reaction mixture, the mixture was extracted with EA (2 × 20 ml), the combined organic layers were washed with saturated NaCl, dried over _{anhydrous Na2SO4} and the solvent was distilled off under reduced pressure to obtain a crude product, which was purified by TLC (eluent: PE/acetone = 5:1) to give a white solid KH025 (70 mg, 60.76%).

^1H NMR (400 MHz, _CDCl3 ) δ 7.72 (d, 1H), 7.58 - 7.49 (m, 1H), 7.42 (d, 1H), 7.29 (d, 1H) , 5.11 - 4.95 (m, 2H), 2.35 (dd, 1H), 1.87 - 1.22 (m, 21H), 1.20 (s, 3H), 0.99 (s , 3H), 0.87 - 0.80 (m, 3H), 0.72 (s, 3H).

[0152] Example 23: Synthesis of compound KH026:

[0153]

[0154] Compound KH026: KHC _- 8 (53 mg, 0.12 mmol) was dissolved in 5 ml of _CH3CN , then _K2CO3 (100 mg, 0.72 mmol) and 2-mercaptobenzoxazole (24 mg, 0.16 mmol) were added. The reaction was carried out at room temperature overnight. TLC monitoring showed that the reaction was complete. 10 ml of water were added to the reaction mixture, the mixture was extracted with EA (2 × 20 ml), the combined organic layers were washed with saturated NaCl, dried over _{anhydrous Na2SO4} and the solvent was distilled off under reduced pressure to obtain a crude product, which was purified by TLC (eluent: PE/acetone = 6:1) to give a white solid KH026 (30 mg, 50.40%).

^1H NMR (400 MHz, CDCl ₃ ) 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).

[0156] Example 24: Synthesis of compound KH027:

[0157]

[0158] Compound KH027: KHC-9 (110 mg, 0.22 mmol) was dissolved in 10 mL of CH ₃ CN, then Cs ₂ CO ₃ (215 mg, 0.66 mmol) and 4-cyanopyrazole (61 mg, 0.66 mmol) were added. The reaction was carried out at room temperature overnight. TLC monitoring showed that the reaction was complete. 10 ml of water were added to the reaction mixture, the mixture was extracted with EA (2 × 20 ml), the combined organic layers were washed with saturated NaCl, dried over anhydrous Na ₂ SO ₄ and the solvent was distilled off under reduced pressure to obtain a crude product, which was purified by TLC (eluent: CH ₂ Cl ₂ /CH ₃ OH = 6:1) to obtain a white solid KH027 (76 mg, 65.30%).

^1H NMR (400 MHz, CDCl ₃ ) 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).

[0160] Example 25: Synthesis of Compound KH028:

[0161]

[0162] Compound KH028: KHC-8 (70 mg, 0.16 mmol) was dissolved in 8 mL of DMSO, then _Cs2CO3 ( ₁₆₀ mg, 0.49 mmol) and 2-hydroxybenzoxazole (66 mg, 0.49 mmol) were added. The reaction was carried out at room temperature overnight. TLC monitoring showed that the reaction was complete. 10 ml of water were added to the reaction mixture, the mixture was extracted with EA (2 × 20 ml), the combined organic layers were washed with saturated NaCl, dried over anhydrous Na ₂ SO ₄ and the solvent was distilled off under reduced pressure to obtain a crude product, which was purified by TLC (eluent: CH ₂ Cl ₂ /CH ₃ OH = 60:1) to obtain a white solid KH028 (55 mg, 71.61%).

^1H NMR (400 MHz, CDCl ₃ ) 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).

[0164] Example 26: Synthesis of compound KH029:

[0165]

[0166] Compound KH029: KHC-8 (60 mg, 0.14 mmol) was dissolved in 5 mL of DMSO, then _Cs2CO3 ( 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. TLC monitoring showed that the reaction was complete. 10 ml of water were added to the reaction mixture, the mixture was extracted with EA (2 × 20 ml), the combined organic layers were washed with saturated NaCl, dried over anhydrous Na ₂ SO ₄ and the solvent was distilled off under reduced pressure to obtain a crude product, which was purified by TLC (eluent: CH ₂ Cl ₂ /CH ₃ OH = 60:1) to give a white solid KH029 (50 mg, 70.72%).

^1H 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).

[0168] Example 27: Synthesis of compound KH030:

[0169]

[0170] Compound KH030: KHC _- 8 (50 mg, 0.12 mmol) was dissolved in 5 mL of DMSO, then _Cs2CO3 (78 mg, 0.24 mmol) and 3-hydroxy-4-methyl-5-methoxy-4H-1,2,4-triazole (31 mg, 0.24 mmol) were added. The reaction was carried out at room temperature overnight. TLC monitoring showed that the reaction was complete. 10 ml of water were added to the reaction mixture, the mixture was extracted with EA (2 × 20 ml), the combined organic layers were washed with saturated NaCl, dried over anhydrous Na ₂ SO ₄ and the solvent was distilled off under reduced pressure to obtain a crude product, which was purified by TLC (eluent: PE/acetone = 4:1) to obtain a white solid KH030 (20 mg, 35.16%).

^1H NMR (400 MHz, CDCl ₃ ) δ 4.47 (k, 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).

[0172] Example 28: Synthesis of Compound KH031:

[0173]

[0174] Compound KH031: KHC-8 (60 mg, 0.14 mmol) was dissolved in 5 mL of CH ₃ CN, then K ₂ CO ₃ (58 mg, 0.42 mmol) and 1-methyl-5-mercapto-1H-tetrazole (68 mg, 0.42 mmol) were added. The reaction was carried out at room temperature. TLC monitoring showed that the reaction was complete. 10 ml of water were added to the reaction mixture, the mixture was extracted with EA (2 × 20 ml), the combined organic layers were washed with saturated NaCl, dried over anhydrous Na ₂ SO ₄ and the solvent was distilled off under reduced pressure to obtain a crude product, which was purified by TLC (eluent: PE/acetone = 4:1) to obtain a white solid KH031 (50 mg, 77.47%).

^1H NMR (400 MHz, CDCl ₃ ) 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).

[0176] Example 29: Synthesis of Compound KH032:

[0177]

[0178] Compound KH032: KHC-8 (50 mg, 0.12 mmol) was dissolved in 5 mL of DMSO, then K ₂ CO ₃ (50 mg, 0.36 mmol) and 1-hydroxy-1H-imidazole (30 mg, 0.36 mmol) were added. The reaction was carried out at room temperature overnight. Monitoring the reaction progress by TLC showed that the reaction was complete. 10 mL of water were added to the reaction mixture, the mixture was extracted with EA (2 × 20 mL), the combined organic layers were washed with saturated NaCl, dried over anhydrous Na ₂ SO ₄ and the solvent was distilled off under reduced pressure to obtain a crude product, which was purified by TLC (eluent: PE/acetone = 2:1) to give a white solid KH032 (11 mg, 21.37%).

^1H NMR (400 MHz, CDCl ₃ ) 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).

[0180] Example 30: Synthesis of Compound KH033:

[0181]

[0182] Compound KH033: KHC-8 (60 mg, 0.14 mmol) was dissolved in 5 ml of DMSO, then _Cs2CO3 ( ₁₃₈ mg, 0.42 mmol) and 2-hydroxyquinoline (61 mg, 0.42 mmol) were added. The reaction was carried out at room temperature overnight. Monitoring the reaction progress by TLC showed that the reaction was complete. 10 ml of water were added to the reaction mixture, the mixture was extracted with EA (2 × 20 ml), the combined organic layers were washed with saturated NaCl, dried over _{anhydrous Na2SO4} and the solvent was distilled off under reduced pressure to obtain a crude product, which was purified by TLC (eluent: PE/acetone = 4:1) to give a white solid KH033 (15 mg, 21.87%).

^1H NMR (400 MHz, CDCl ₃ ) 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).

[0184] Example 31: Synthesis of compound KH034:

[0185]

[0186] Compound KH034: KHC-8 (60 mg, 0.14 mmol) was dissolved in 5 mL of DMSO, then K ₂ CO ₃ (58 mg, 0.42 mmol) and 3-hydroxy-1H-1,2,4-triazole (36 mg, 0.42 mmol) were added. The reaction was carried out at room temperature overnight. Monitoring the reaction progress by TLC showed that the reaction was complete. 10 ml of water were added to the reaction mixture, the mixture was extracted with EA (2 × 20 ml), the combined organic layers were washed with saturated NaCl, dried over anhydrous Na ₂ SO ₄ and the solvent was distilled off under reduced pressure to obtain a crude product, which was purified by TLC (eluent: CH ₂ Cl ₂ /CH ₃ OH = 40:1) to obtain a white solid KH034 (40 mg, 66.45%).

^1H NMR (400 MHz, CDCl ₃ ) δ 9.30 (s, 1H), 7.42 (s, 1H), 4.47 (k, 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).

[0188] Example 32: Synthesis of compound KH035:

[0189]

[0190] Compound KH035: KHC-8 (80 mg, 0.19 mmol) was dissolved in 6 mL of DMSO, then _Cs2CO3 ( ₁₈₄ mg, 0.56 mmol) and 8-fluoro-4-hydroxyquinazoline (92 mg, 0.56 mmol) were added. The reaction was carried out at room temperature overnight. Monitoring the reaction progress by TLC showed that the reaction was complete. 10 ml of water were added to the reaction mixture, the mixture was extracted with EA (2 × 20 ml), the combined organic layers were washed with saturated NaCl, dried over anhydrous Na ₂ SO ₄ and the solvent was distilled off under reduced pressure to obtain a crude product, which was purified by TLC (eluent: PE/acetone = 2:1) to obtain a white solid KH035 (65 mg, 67.21%).

^1H NMR (400 MHz, CDCl ₃ ) 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).

[0192] Example 33: Synthesis of Compound KH036:

[0193]

[0194] Compound KH036: KHC-8 (60 mg, 0.14 mmol) was dissolved in 5 mL of DMSO, then K ₂ CO ₃ (58 mg, 0.42 mmol) and 6-fluoropyridin[3,4-d]pyrimidin-4-one (69 mg, 0.42 mmol) were added. The reaction was carried out at room temperature overnight. Monitoring the reaction progress by TLC showed that the reaction was complete. 10 ml of water were added to the reaction mixture, the mixture was extracted with EA (2 × 20 ml), the combined organic layers were washed with saturated NaCl, dried over anhydrous Na ₂ SO ₄ and the solvent was distilled off under reduced pressure to obtain a crude product, which was purified by TLC (eluent: CH ₂ Cl ₂ /CH ₃ OH = 40:1) to give a white solid KH036 (40 mg, 56.02%).

^1H NMR (400 MHz, CDCl ₃ ) δ 8.82 (s, 1H), 7.80 (s, 1H), 7.66 (d, 1H), 4.80 (k, 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).

[0196] Example 34: Synthesis of Compound KH037:

[0197]

[0198] Compound KH037: KHC-8 (60 mg, 0.14 mmol) was dissolved in 5 mL of CH ₃ CN, then K ₂ CO ₃ (58 mg, 0.42 mmol) and 4-methyl-4H-3-mercapto-1,2,4-triazole (49 mg, 0.42 mmol) were added. The reaction was carried out at room temperature overnight. Monitoring the reaction progress by TLC showed that the reaction was complete. 10 ml of water were added to the reaction mixture, the mixture was extracted with EA (2x20 ml), the combined organic layers were washed with saturated NaCl, dried over _{anhydrous Na2SO4} and the solvent was distilled off under reduced pressure to obtain a crude product, which was purified by TLC (eluent: PE/acetone=3:2) to obtain a white solid KH037 (53 mg, 82.30%).

^1H NMR (400 MHz, CDCl ₃ ) 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).

[0200] Example 35: Synthesis of Compound KH038:

[0201]

[0202] Compound KH038: KHC-8 (60 mg, 0.14 mmol) was dissolved in 5 ml of DMSO, 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. Monitoring the reaction progress by TLC showed that the reaction was complete. 10 ml of water were added to the reaction mixture, the mixture was extracted with EA (2 × 20 ml), the combined organic layers were washed with saturated NaCl, dried over _{anhydrous Na2SO4} and the solvent was distilled off under reduced pressure to obtain a crude product, which was purified by TLC (eluent: PE/acetone = 15:1) to give a white solid KH038 (39 mg, 65.00%).

^1H NMR (300 MHz, CDCl ₃ ) 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).

[0204] Example 36: Synthesis of Compound KH039:

[0205]

[0206] Compound KH039: KHC-8 (75 mg, 0.18 mmol) was dissolved in 5 ml of _CH3CN , then _Cs2CO3 ( ₁₇₁ mg, 0.53 mmol) and 5-phenyltetrazole (77 mg, 0.53 mmol) were added. The reaction was carried out at room temperature overnight. Monitoring the reaction progress by TLC showed that the reaction was complete. 10 ml of water were added to the reaction mixture, the mixture was extracted with EA (2 × 20 ml), the combined organic layers were washed with saturated NaCl, dried over _{anhydrous Na2SO4} and the solvent was distilled off under reduced pressure to obtain a crude product, which was purified by TLC (eluent: PE/acetone = 3:1) to give a white solid KH039 (79 mg, 89.39%).

^1H NMR (400 MHz, CDCl ₃ ) 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).

[0208] Example 37: Synthesis of Compound KH040:

[0209]

[0210] Compound KH040: KHC _- 8 (60 mg, 0.14 mmol) was dissolved in 5 mL of _CH3CN , then _Cs2CO3 (91 mg, 0.28 mmol) and 4-cyanopyrazole (26 mg, 0.28 mmol) were added. The reaction was carried out at room temperature overnight. Monitoring the reaction progress by TLC showed that the reaction was complete. 10 ml of water were added to the reaction mixture, the mixture was extracted with EA (2 × 20 ml), the combined organic layers were washed with saturated NaCl, dried over anhydrous Na ₂ SO ₄ and the solvent was distilled off under reduced pressure to obtain a crude product, which was purified by TLC (eluent: CH ₂ Cl ₂ /CH3OH = 40:1) to give a white solid KH040 (16 mg, 26.09%). ^1H NMR (400 MHz, CDCl ₃ ) δ 7.82 (s, 1H), 7.81 (s, 1H), 5.00 (k, 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).

[0212] Example 38: Synthesis of compound KH041:

[0213]

[0214] Compound KH041: KHC-8 (70 mg, 0.16 mmol) was dissolved in 5 mL of CH ₃ CN, then Cs ₂ CO ₃ (160 mg, 0.49 mmol) and 2-mercaptothiophene (57 mg, 0.49 mmol) were added. The reaction was carried out at room temperature overnight. Monitoring the reaction progress by TLC showed that the reaction was complete. 10 ml of water were added to the reaction mixture, the mixture was extracted with EA (2 × 20 ml), the combined organic layers were washed with saturated NaCl, dried over anhydrous Na ₂ SO ₄ and the solvent was distilled off under reduced pressure to obtain a crude product, which was purified by TLC (eluent: CH ₂ Cl ₂ /PE = 1:1) to give a gray solid KH041 (27 mg, 35.53%). ^1H NMR (400 MHz, CDCl ₃ ) 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).

[0216] Example 39: Synthesis of compound KH042:

CT
[0217]

[0218] Compound KH042: KHC-8 (150 mg, 0.35 mmol) was dissolved in 15 mL of water and 30 mL of acetone, then K ₂ CO ₃ (49 mg, 0.35 mmol) was added. The reaction was carried out at room temperature overnight. Monitoring the reaction progress by TLC showed that the reaction was complete. 10 mL of water was added to the reaction mixture, the mixture was extracted with EA (2 × 30 mL), the combined organic layers were washed with saturated NaCl, dried over anhydrous Na ₂ SO ₄ and the solvent was distilled off under reduced pressure to obtain a crude product, which was purified by TLC (eluent: CH ₂ Cl ₂ /CH ₃ OH = 50:1) to give a white solid KH042 (21 mg, 16.54%). ^1H 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).

[0220] Example 39: Synthesis of compound KH043 and KH044:

CT
[0221]

[0222] Compounds KH043 and KH044: KHC-8 (60 mg, 0.14 mmol) was dissolved in 5 mL of CH ₃ CN, then Cs ₂ CO ₃ (91 mg, 0.28 mmol) and 1H-triazole (15 mg, 0.21 mmol) were added. The reaction was carried out at room temperature overnight. Monitoring the reaction progress by TLC showed that the reaction was complete. To the reaction mixture were added 10 ml of water, the mixture was extracted with EA (2 × 20 ml), the combined organic layers were washed with saturated NaCl, dried over _{anhydrous Na2SO4} and the solvent was distilled off under reduced pressure to obtain a crude product, which _was purified by TLC (eluent: _CH2Cl2 / _CH3OH =40:1) to give a white solid KH6070100-1 (12 mg, 20.70%) and a white solid KH6070100-2 (27 mg, 46.58%), respectively.

^1H NMR (400 MHz, CDCl ₃ ) 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).

^1H 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).

[0225] Example 40: Synthesis of compound KH045:

[0226] KHC-11 Compound: KHC-10 (150.0 g, 183.8 mmol) was dissolved in 500 mL of THF, and 10% Pd/C (6.0 g) was added to the solution. The solution was then hydrogenated under H ₂ atmosphere and continued to stir at RT for 24 h, and then filtered. The filtrate was concentrated under vacuum to obtain a crude product, which was then stirred with a solvent (eluent: PE: acetone = 1: 1) to obtain KHC-11 (150.0 g, yield 99.26%).

[0227] KHC-12 compound: KHC-11 (150.0 g, 547.4 mmol) was dissolved in 1000 mL of _CH3OH , then I2 (14.0 g, 54.8 mmol) was added to the solution, the solution was heated to 60° and continued to stir for 12 hours, then it was concentrated under vacuum and the residue was purified by flash chromatography on a silica gel column (eluent: PE:EA=10:1) to obtain a white solid KHC-12 (130.0 g, 74.20%).

[0228] KHC-13 Compound: KHC-12 (130.0 g, 406.2 mmol) was dissolved in 1000 mL DMF and t-BuOK (91.0 g, 812.4 mmol), trimethyliodonium sulfide (165.8 g, 812.7 mmol) were added to the solution, the solution was continued to stir at RT for 12 h under N ₂ atmosphere, then brine was added to the reaction mixture, 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%).

[0229] KHC-14 compound: KHC-13 (120.0 g, 359.4 mmol) was dissolved in 900 mL of ethanol and 160 mL of water. NaN ₃ (70.2 g, 1077.9 mmol) and NH ₄ Cl (67.2 g, 1257.9 mmol) were added to the solution, and the solution was continued to stir at 90 °C overnight. Brine was added to the reaction mixture, the mixture was extracted with EA (3 × 1 L), and the organic layers were combined and concentrated in vacuo. The residue was purified by flash column chromatography on silica gel (eluent: PE:EA=10:1) to give a white solid KHC-14 (130.0 g, 95.59%).

[0230] KHC-15: KHC-14 (125.6 g, 332.4 mmol) was dissolved in 1000 mL of CH ₃ CN, NaI (249.3 g, 1662.0 mmol) was added to the solution, the solution was continued to stir at RT for half an hour, then trimethylchlorosilane (TMSCl) (144.9, 1329.6 mmol) was added dropwise to the reaction solution and the reaction solution was continued to stir at RT for 4 hours. Brine was added to the reaction mixture, the mixture was extracted with EA (3 × 1 L), the organic layers were combined and concentrated in vacuo. The residue was purified by flash column chromatography on silica gel (eluent: PE:EA=10:1) to give a white solid KHC-15 (72.0 g, 75.20%).

[0231] KHC-16 compound: KHC-15 (72.0 g, 250.0 mmol) was dissolved in 1000 mL of _CH3OH , I2 (19.0 g, 75.0 mmol) was added to the solution, the reaction solution was heated to 60° and continued to stir for 12 hours, then the reaction mixture was concentrated under vacuum, the residue was purified by flash chromatography on a silica gel column (eluent: PE:EA=10:1) to give a white solid KHC-16 (53.0 g, 63.50%).

[0232] KHC-17 compound: Trimethylsilylacetylene (77.8 g, 793.0 mmol) in 1000 mL of THF was added, n-butyl lithium solution (1.6 M, 329.0 mmol, 205.6 mL) was added at -78°, and the reaction solution was continued to stir for 2 hours. Then, 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° for 2 hours and gradually warmed to 0°, then it was poured into a saturated NH ₄ Cl solution, and the mixture was extracted with EA (800 mL×3). The organic layers were combined and concentrated in vacuo, and the residue was purified by column chromatography (eluent: PE:EA = 10:1) to give a white solid KHC-17 (56.0 g, 81.55%).

[0233] KHC-18: KHC-17 (56.0 g, 129.3 mmol) in 800 mL of THF was mixed with 1.0 M tetrabutylammonium fluoride (TBAF) (129.3 mmol, 129.3 mL) at 0° and the mixture was continued to stir for 1 h. Brine was added to the reaction mixture and the mixture was extracted with EA (3 × 1 L). The organic layers were combined and concentrated in vacuo, the residue was purified by flash column chromatography on silica gel (eluent: PE:EA=5:1) to give a white solid KHC-18 (43.0 g, 92.30%).

[0234] KHC-19: KHC-18 (38.0 g, 105.6 mmol) in 800 mL THF was mixed with 1.0 M HCl until pH=3 and the mixture was continued stirring at RT for 12 h. Brine (1.0 L) was added to the reaction mixture and the mixture was extracted with EA (3×1 L). The organic layers were combined and concentrated in vacuo. The residue was then purified by flash column chromatography on silica gel (eluent: PE:EA=5:1) to give a white solid KHC-19 (32.0 g, 96.50%).

[0235] KHC-20 Compound: Methanesulfonic acid (148.0 g, 1541.6 mmol) was mixed with 1000 mL _{of CH2Cl2} , KHC-19 (32.0 g, 101.9 mmol) in 1000 mL _{of CH2Cl2} was added dropwise to the solution, the reaction solution was continued to stir at 60 °C for 2 h and cooled to RT. Brine (1.0 L) was added to the reaction mixture, the mixture _was extracted with _CH2Cl2 (3 × 1.0 L). The organic layers were combined and concentrated in vacuo. The residue was purified by flash column chromatography on silica gel (eluent: PE:EA=10:1) to give a white solid KHC-20 (20.0 g, 62.50%).

[0236] 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 H ₂ atmosphere. The reaction solution was continued to stir at RT for 2 days and then filtered. The filtrate was concentrated under vacuum to obtain a crude product, which was then purified by flash column chromatography on silica gel (eluent: PE:EA=15:1) to give a white solid KHC-21 (17.0 g, 84.40%).

[0237] Compound KH045: To a mixture of _FeCl3 (23.0 g, 142.0 mmol) and LiCl (13.5 g, 321.4 mmol) in a 2000 mL three-neck flask under _N2 atmosphere was added 800 mL of anhydrous THF, the mixture was continued to stir at RT for 20 min and then cooled to -40°C, _CH3MgBr (3 M, 190 mL, 570.0 mmol) was added dropwise to the solution, then the solution was continued to stir at -40°C for 30 min. KHC-21 (17.0 g, 53.8 mmol) in anhydrous THF (200 mL) was added dropwise to the above reaction solution, then the solution was warmed to -20°C and continued to stir for 4 h. Monitoring of the reaction progress by TLC showed that the reaction was complete. The reaction solution was quenched with saturated _NH4Cl (40 mL) and the mixture was extracted with EA (3×1.0 L). The combined organic layers were washed with saturated brine l, dried over _{anhydrous Na2SO4} and concentrated in vacuo to give the crude product, which 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%).

^1H NMR (400 MHz, CDCl ₃ ) 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).

[0239]

[0240] Example 41: Synthesis of Compound KH046:

[0241]

[0242] Compound KHC-22 and Compound KHC-23: Compound KH045 (1.50 g, 4.50 mmol) was dissolved in 40 mL of _CH3OH , and then _Br2 (1.08 g, 6.75 mmol) was added dropwise to the solution. The reaction solution turned from orange to light yellow at RT within 4 h, and monitoring the reaction progress by TLC showed that the reaction was complete. 30 mL of water were added to the reaction mixture, the mixture was extracted with EA (2 × 65 mL), the combined organic layers were washed with saturated NaCl, dried over _{anhydrous Na2SO4} , and the solvent was distilled off under reduced pressure to obtain a crude product, which was purified by flash chromatography on silica gel (eluent: _CH2Cl2 ) to give a white solid KHC- ₂₂ (1.19 g, 64.34%) and a white solid KHC-23 (0.33 g, 17.84%).

[0243] Compound KH046: KHC-22 (50 mg, 0.12 mmol) was dissolved in 5 mL of DMSO, then K ₂ CO ₃ (21 mg, 0.15 mmol) and 8-fluoro-4-hydroxyquinazoline (25 mg, 0.15 mmol) were added. The reaction was carried out at room temperature overnight. Monitoring the reaction progress by TLC showed that the reaction was complete. 10 ml of water were added to the reaction mixture, the mixture was extracted with EA (2 × 20 ml), the combined organic layers were washed with saturated NaCl, dried over anhydrous Na ₂ SO ₄ , and the solvent was distilled off under reduced pressure to obtain a crude product, which was purified by TLC (eluent: PE/acetone = 4:1) to give a white solid KH046 (33 mg, 55.56%).

^1H NMR (400 MHz, CDCl ₃ ) δ 8.06 (d, 1H), 7.88 (s, 1H), 7.55 - 7.40 (m, 2H), 4.80 (k, 2H) , 2.44 (dd, 1H), 2.04 - 2.01 (m, 1H), 1.91 - 1.88 (m, 1H), 1.84 - 1.16 (m, 21H), 1 .15 - 0.80 (m, 8H).

[0245] Example 42: Synthesis of Compound KH047

[0246]

[0247] Compound KH047: KHC-22 (50 mg, 0.12 mmol) was dissolved in 5 mL of CH ₃ CN, then Cs ₂ CO ₃ (49 mg, 0.15 mmol) and 5-mercapto-1-methyl-1H-tetrazole (17 mg, 0.15 mmol) were added. The reaction was carried out at room temperature overnight. Monitoring the reaction progress by TLC showed that the reaction was complete. 10 ml of water were added to the reaction mixture, the mixture was extracted with EA (2 × 20 ml), the combined organic layers were washed with saturated NaCl, dried over anhydrous Na ₂ SO ₄ and the solvent was distilled off under reduced pressure to obtain a crude product, which was purified by TLC (eluent: PE/acetone = 4:1) to give a white solid KH047 (51 mg, 50.12%).

^1H NMR (400 MHz, CDCl ₃ ) 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).

[0249] Example 43: Synthesis of Compound KH048

[0250]

[0251] Compound KH048: KHC-22 (60 mg, 0.14 mmol) was dissolved in 5 mL of DMSO, then K ₂ CO ₃ (58 mg, 0.42 mmol) and 6-fluoropyridin[3,4-d]pyrimidin-4-one (69 mg, 0.42 mmol) were added. The reaction was carried out at room temperature overnight. Monitoring the reaction progress by TLC showed that the reaction was complete. 10 ml of water were added to the reaction mixture, the mixture was extracted with EA (2 × 20 ml), the combined organic layers were washed with saturated NaCl, dried over anhydrous Na ₂ SO ₄ and the solvent was distilled off under reduced pressure to obtain a crude product, which was purified by TLC (eluent: PE/CH ₃ OH = 40:1) to give a white solid KH048 (37 mg, 50.12%).

^1H 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).

[0253] Example 44: Synthesis of Compound KH049

[0254]

[0255] Compound KH049: KHC-22 (60 mg, 0.15 mmol) was dissolved in 5 mL of CH ₃ CN, then Cs ₂ CO ₃ (62 mg, 0.19 mmol) and 2-mercaptobenzoxazole (29 mg, 0.19 mmol) were added. The reaction was carried out at room temperature overnight. Monitoring the reaction progress by TLC showed that the reaction was complete. 10 ml of water were added to the reaction mixture, the mixture was extracted with EA (2 × 20 ml), the combined organic layers were washed with saturated NaCl, dried over anhydrous Na ₂ SO ₄ and the solvent was distilled off under reduced pressure to obtain a crude product, which was purified by TLC (eluent: PE/acetone = 4:1) to give a white solid KH049 (42 mg, 42.92%).

^1H NMR (400 MHz, CDCl ₃ ) 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).

[0257] Example 45: Synthesis of Compound KH050

[0258]

[0259] Compound KH050: KHC-22 (60 mg, 0.15 mmol) was dissolved in 5 mL of CH ₃ CN, then Cs ₂ CO ₃ (62 mg, 0.19 mmol) and 3-mercapto-4-methyl-4H-1,2,4-triazole (22 mg, 0.19 mmol) were added. The reaction was carried out at room temperature overnight. Monitoring the reaction progress by TLC showed that the reaction was complete. 10 ml of water were added to the reaction mixture, the mixture was extracted with EA (2 × 20 ml), the combined organic layers were washed with saturated NaCl, dried over anhydrous Na ₂ SO ₄ and the solvent was distilled off under reduced pressure to obtain a crude product, which was purified by TLC (eluent: CH ₂ Cl ₂ /CH ₃ OH = 70:1) to give a white solid KH050 (17 mg, 25.41%).

^1H NMR (400 MHz, CDCl ₃ ) 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).

[0261] Example 46: Synthesis of Compound KH051

[0262]

[0263] Compound KH051: KHC-22 (60 mg, 0.15 mmol) was dissolved in 5 mL of DMSO, then K ₂ CO ₃ (58 mg, 0.42 mmol) and 3-hydroxy-1H-1,2,4-triazole (36 mg, 0.42 mmol) were added. The reaction was carried out at room temperature overnight. Monitoring the reaction progress by TLC showed that the reaction was complete. 10 ml of water were added to the reaction mixture, the mixture was extracted with EA (2 × 20 ml), the combined organic layers were washed with saturated NaCl, dried over anhydrous Na ₂ SO ₄ and the solvent was distilled off under reduced pressure to obtain a crude product, which was purified by TLC (eluent: CH ₂ Cl ₂ /CH ₃ OH = 40:1) to obtain a white solid KH051 (27 mg, 46.36%).

^1H NMR (400 MHz, _CDCl3 ) δ 9.57 (s, 1H), 7.42 (s, 1H), 4.46 (k, 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).

[0265] Example 47: Synthesis of Compound KH052 and Compound KH053

[0266]

[0267] Compound KH052 and compound KH053: KHC-22 (100 mg, 0.24 mmol) were dissolved in 5 mL of CH ₃ CN, then Cs ₂ CO ₃ (234 mg, 0.72 mmol) and 1H-tetrazole (50 mg, 0.72 mmol) were added. The reaction was carried out at room temperature overnight. Monitoring the reaction progress by TLC showed that the reaction was complete. To the reaction mixture were added 10 ml of water, the mixture was extracted with EA (2 × 20 ml), the combined organic layers were washed with saturated NaCl, dried over _{anhydrous Na2SO4} and the solvent _was distilled off under reduced pressure to obtain a crude product, which was purified by TLC (eluent: _CH2Cl2 / _CH3OH =30:1) to give a white solid KH6070069-1 (19 mg, 19.74%) and a white solid KH6070069-2 (44 mg, 45.72%), respectively.

^1H 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).

^1H NMR (400 MHz, CDCl ₃ ) 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).

[0270] Example 48: Synthesis of Compound KH054

[0271]

[0272] Compound KH054: KHC-22 (60 mg, 0.15 mmol) was dissolved in 5 mL of CH ₃ CN, then Cs ₂ CO ₃ (62 mg, 0.19 mmol) and 2-hydroxyoxazole[4,5-b]pyridine (26 mg, 0.19 mmol) were added. The reaction was carried out at room temperature overnight. Monitoring the reaction progress by TLC showed that the reaction was complete. 10 ml of water were added to the reaction mixture, the mixture was extracted with EA (2 × 20 ml), the combined organic layers were washed with saturated NaCl, dried over anhydrous Na ₂ SO ₄ and the solvent was distilled off under reduced pressure to obtain a crude product, which was purified by TLC (eluent: CH ₂ Cl ₂ /CH ₃ OH = 70:1) to obtain a white solid KH054 (17 mg, 42.92%).

^1H NMR (400 MHz, CDCl ₃ ) δ 7.25 (d, 1H), 7.01 (d, 1H), 6.69 (t, 1H), 5.11 (k, 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).

[0274] Example 49: Synthesis of Compound KH055

[0275]

[0276] Compound KH055: KHC-22 (60 mg, 0.15 mmol) was dissolved in 5 mL of CH ₃ CN, then Cs ₂ CO ₃ (156 mg, 0.48 mmol) and 4-cyanopyrazole (45 mg, 0.48 mmol) were added. The reaction was carried out at room temperature overnight. Monitoring the reaction progress by TLC showed that the reaction was complete. 10 ml of water were added to the reaction mixture, the mixture was extracted with EA (2 × 20 ml), the combined organic layers were washed with saturated NaCl, dried over anhydrous Na ₂ SO ₄ and the solvent was distilled off under reduced pressure to obtain a crude product, which was purified by TLC (eluent: CH ₂ Cl ₂ /CH ₃ OH = 30:1) to give a white solid KH055 (51 mg, 50.12%).

^1H NMR (400 MHz, _CDCl3 ) δ 7.83 (s, 1H), 7.81 (s, 1H), 4.99 (k, 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).

[0278] Example 50: Synthesis of Compound KH056

[0279]

[0280] Compound KH056: KHC-23 (40 mg, 0.10 mmol) was dissolved in 5 mL of CH ₃ CN, then Cs ₂ CO ₃ (198 mg, 0.3 mmol) and 4-cyanopyrazole (28 mg, 0.30 mmol) were added. The reaction was carried out at room temperature overnight. Monitoring the reaction progress by TLC showed that the reaction was complete. 10 ml of water were added to the reaction mixture, the mixture was extracted with EA (2 × 20 ml), the combined organic layers were washed with saturated NaCl, dried over anhydrous Na ₂ SO ₄ and the solvent was distilled off under reduced pressure to obtain a crude product, which was purified by TLC (eluent: CH ₂ Cl ₂ /CH ₃ OH = 30:1) to give a white solid KH056 (12 mg, 23.30%).

^1H NMR (400 MHz, CDCl ₃ ) 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).

[0282] Example 51: Synthesis of Compound KH057

[0283]

[0284] Compound KH057: KHC-22 (60 mg, 0.15 mmol) was dissolved in 5 ml of DMSO, then _Cs2CO3 ( ₁₄₃ mg, 0.44 mmol) and 2-hydroxybenzoxazole (59 mg, 0.44 mmol) were added. The reaction was carried out at room temperature overnight. Monitoring the reaction progress by TLC showed that the reaction was complete. 10 ml of water were added to the reaction mixture, the mixture was extracted with EA (2 × 20 ml), the combined organic layers were washed with saturated NaCl, dried over _{anhydrous Na2SO4} and the solvent was distilled off under reduced pressure to obtain a crude product, which was purified by TLC (eluent: PE/acetone = 3:1) to give a white solid KH057 (30 mg, 42.92%).

^1H NMR (400 MHz, CDCl ₃ ) 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).

[0286] Example 52: Synthesis of Compound KH058

[0287]

[0288] Compound KH058: KHC-22 (80 mg, 0.19 mmol) was dissolved in 5 mL of DMSO, then _Cs2CO3 ( ₁₉₀ mg, 0.58 mmol) and 8-methyl-4-quinazolinone (93 mg, 0.58 mmol) were added. The reaction was carried out at room temperature overnight. Monitoring the reaction progress by TLC showed that the reaction was complete. 10 ml of water were added to the reaction mixture, the mixture was extracted with EA (2 × 20 ml), the combined organic layers were washed with saturated NaCl, dried over anhydrous Na ₂ SO ₄ and the solvent was distilled off under reduced pressure to obtain a crude product, which was purified by TLC (eluent: PE/acetone = 3:1) to give a white solid KH058 (28 mg, 30.01%).

^1H 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).

[0290] Example 53: Synthesis of Compound KH059

[0291]

[0292] Compound KH059: KHC-22 (80 mg, 0.19 mmol) was dissolved in 5 mL of DMSO, then _Cs2CO3 ( ₁₉₀ mg, 0.58 mmol) and 8-hydroxy-1,7-naphthyridine (85 mg, 0.58 mmol) were added. The reaction was carried out at room temperature overnight. Monitoring the reaction progress by TLC showed that the reaction was complete. 10 ml of water were added to the reaction mixture, the mixture was extracted with EA (2 × 20 ml), the combined organic layers were washed with saturated NaCl, dried over anhydrous Na ₂ SO ₄ and the solvent was distilled off under reduced pressure to obtain a crude product, which was purified by TLC (eluent: PE/acetone = 3:1) to give a white solid KH058 (28 mg, 30.89%).

^1H 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).

[0294] Example 54: Synthesis of Compound KH060

[0295]

[0296] Compound KH060: KHC-22 (100 mg, 0.24 mmol) was dissolved in 5 mL of DMSO, then _Cs2CO3 ( ₁₅₆ mg, 0.48 mmol) and 3-hydroxy-4-methyl-5-methoxy-4H-1,2,4-triazole (62 mg, 0.48 mmol) were added. The reaction was carried out at room temperature overnight. Monitoring the reaction progress by TLC showed that the reaction was complete. 10 ml of water were added to the reaction mixture, the mixture was extracted with EA (2 × 20 ml), the combined organic layers were washed with saturated NaCl, dried over anhydrous Na ₂ SO ₄ and the solvent was distilled off under reduced pressure to obtain a crude product, which was purified by TLC (eluent: PE/acetone = 4:1) to give a white solid KH058 (56 mg, 50.72%).

^1H NMR (400 MHz, CDCl ₃ ) 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).

[0298] Example 55: Synthesis of Compound KH061

[0299]

[0300] Compound KH061: KHC-22 (80 mg, 0.19 mmol) was dissolved in 5 ml of DMSO, then _Cs2CO3 ( ₁₅₆ mg, 0.48 mmol) and 2-hydroxyquinoline (84 mg, 0.58 mmol) were added. The reaction was carried out at room temperature overnight. Monitoring the reaction progress by TLC showed that the reaction was complete. 10 ml of water were added to the reaction mixture, the mixture was extracted with EA (2 × 20 ml), the combined organic layers were washed with saturated NaCl, dried over _{anhydrous Na2SO4} and the solvent was distilled off under reduced pressure to obtain a crude product, which was purified by TLC (eluent: PE/acetone = 4:1) to give a white solid KH058 (45 mg, 49.76%).

^1H NMR (400 MHz, CDCl ₃ ) 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).

[0302] Example 56: In vitro cell viability assay

[0303] The control compound used in the present invention has the chemical structure shown below:

[0304]

[0305] The specific process for synthesizing the control compound was carried out according to CN105339381B.

[0306] The present invention utilizes two subunits of the recombinant GABA _A receptor, α ₁ β ₂ γ ₂ and α ₄ β ₃ δ, to evaluate the efficacy of representative compounds in vitro using the electrophysiological patch-clamp technique.

[0307] 56.1 Solution for electrophysiological study

[0308] Compositions of extracellular and intracellular fluid.

[0309] Extracellular fluid: 140 mM NaCl, 5 mM CsCl, 2 mM CaCl ₂ 2H ₂ O, 1 mM MgCl ₂ 6H ₂ O, 5 mM HEPES, 10 mM D-glucose, pH=7.4 (NaOH).

[0310] Intracellular fluid: 130 mM CsCl, 0.1 mM CaCl2 H ₂ O, 2 mM MgCl ₂ 6H ₂ O, 1.1 mM ethylene glycol tetraacetic acid (EGTA), 5 mM Na ₂ -ATP, 10 mM HEPES, pH=7.2 (CsOH).

[0311] The resulting intracellular fluid was divided into 1 ml aliquots per tube, frozen and stored in a freezer at -20°, and freshly thawed intracellular fluid was used in the experiment every day. All intracellular fluid was consumed within three months.

[0312] 56.2 Cell lines with stable expression of ion channels

[0313]

Ion channel Expressed gene Gene information Cell line GABA _A1
(α1β2γ2) GABRA1
GABRB2
GABRG2 NM_000806
NM_021911
NM_198904 HEK293 GABA _A4
(α4β3δ) GABRA4
GABRB3
GABRD NM_000809
NM_000814
NM_000815 HEK293

[0314] 56.3 Cell culture

[0315] 56.3.1 Cell culture with GABA _A (α1β2γ2)

[0316] HEK293 cells stably expressing GABA _A (α1β2γ2) receptors 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 humidity-controlled incubator at 37° with 5% carbon dioxide.

[0317] Cell passage: The old medium was removed and the cells were washed once with PBS. Then 1 ml of 0.25% trypsin-EDTA solution was added and incubated at 37° for 0.5 min. When the cells detached from the bottom of the dish, 5 ml of complete medium pre-warmed at 37° was added. Cell aggregates were separated by gentle pipetting. The cell suspension was transferred to a sterile centrifuge tube, and then the cell suspension was centrifuged at 1000 rpm for 5 min to collect the cells. Cells were expanded or maintained by seeding 2.5 x ¹⁰⁵ cells in a 6 cm cell culture dish (final medium volume: 5 ml).

[0318] To maintain electrophysiological activity, the cell density should not exceed 80%.

[0319] Before manual spot-clamp analysis, cells were detached using 0.25% trypsin-EDTA. Then, 8× ¹⁰³ cells were seeded on coverslips, cultured in a 24-well plate (final medium volume: 500 μl) and analyzed after 18 h by spot-clamp analysis.

[0320] 56.3.2 Cell culture with GABA _A (α4β3δ)

[0321] The HEK293 cell line was cultured in DMEM containing 10% fetal bovine serum, 5% carbon dioxide and 37°C.

[0322] Cell passage: The old medium was removed and the cells were washed once with PBS. Then, 1 ml of 0.25% trypsin-EDTA solution was added to the treated cells. The culture dish was incubated at 37°C in a pre-warmed incubator for 0.5 min. Once the cells were detached, approximately 5 ml of complete medium pre-warmed at 37°C was added. Cell aggregates were separated by gentle pipetting. The cell suspension was transferred to a sterile centrifuge tube, and then the cell suspension was centrifuged at approximately 1000 rpm for 5 min. Cells were expanded or maintained by seeding 2.5 x 10 ⁵ cells into a 6 cm dish (final medium volume: 5 ml). For transfection, 8× ¹⁰³ cells were seeded on coverslips and cultured in a 24-well plate (final medium volume: 5 ml).

[0323] Transfection: On day 2, GABA _A4 transfection was performed (1:1:1 ratio of plasmids α4, β3, δ). The X-tremeGENE HP DAN Transfection Reagent Kit was used. 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 the amount of transfection reagent was 1 μl. The following specific steps were performed to prepare 12 wells. 600 μl of Opti-MEM was added to a sterile centrifuge tube along with 6 μg of plasmid and allowed to mix well; Then 12 µl of transfection reagent were added, mixed well, followed by incubation for 15 min at room temperature. The transfection complex was then inoculated into the cells, 50 µl per well, and the transfection complex was gently vortexed and mixed.

[0324] This mixture was replaced on the third day and spot-clamp analysis was performed on the fourth day.

[0325] 56.4 Local potential clamp analysis

[0326] A glass pipette (BF 150-86-10, Sutter Instruments) was pulled using a micropipette puller (P97, Sutter Instruments). The glass pipette was controlled using a micromanipulator (MP285, Sutter Instruments) under a microscope (IX71, Olympus). After touching the cell, a slight negative pressure was applied to the pipette to achieve a high contact resistance (GΩ). Fast capacitance compensation (in pF) was performed after a high tight contact was established and the membrane was ruptured. Fast capacitance compensation (in pF) of the cell was performed as a result of whole-cell capacitance compensation after a “whole cell” configuration was achieved. No leakage subtraction was performed.

[0327] The test and control solutions were poured into a recording chamber mounted on the stage of an inverted microscope using a gravity-fed solution delivery system. During the experiment, the solutions were removed from the recording chamber by vacuum aspiration. Each concentration was tested multiple times. All tests were performed at room temperature.

[0328] In the whole-cell model _{, current through GABA A1} receptors was recorded using the experimental protocol described below. After establishing tight contact, the cell was held at -70 mV in the non-breaking mode to record the peak current. 3 μM GABA was added to the cell to activate the current. After adding test compounds, 3 μM GABA was added to each concentration of test compounds for 30 s, followed by a 2-min wash 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 software (HEKA).

[0329] In the whole-cell model _{, current through GABA A4} receptors was recorded using the experimental protocol described below. After establishing tight contact, the cell was held at -70 mV in the non-breaking mode to record the peak current. 10 nM GABA was added to the cell to activate the current. After adding test compounds, 10 nM GABA was added to each concentration of test compounds for 30 s, followed by a 2-min wash 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 software (HEKA).

[0330] 56.5 Data Quality Standard

[0331] The following criteria were used to determine whether the results were acceptable:

[0332] (1) Electrode resistance < 5 MOhm

[0333] (2) Contact resistance > 1 GΩ

[0334] (3) Access resistance at start < 15 Mohm

[0335] (4) Access resistance at end < 15 Mohm

[0336] (5) There is no obvious spontaneous decay of current.

[0337] (6) There is no apparent leakage current when the membrane potential is -70 mV.

[0338] 56.6 Analysis of Results

[0339] For GABA _A1 detection, the currents from 3 μM GABA and various concentrations of test compounds, respectively, were normalized to the current of 100 μM GABA (peak compound current)/(peak medium current), and then the activation rate corresponding to each concentration was calculated. For GABA _A4 detection, the currents from 10 nM GABA and various concentrations of test compounds, respectively, were normalized to the current of 10 μM GABA (peak compound current)/(peak medium current), and then the activation rate corresponding to each concentration was calculated. The mean values, standard deviation, and standard error were calculated for each test group. The EC50 value was calculated using nonlinear regression:

Y=at the bottom of the plateau + (at the top of the plateau - at the bottom of the plateau)/(1+10^((LogEC50-X) ×HillSlope))

To fit the data to a concentration-response curve, EC50 was the concentration at which the half-maximal effect was observed and HillSlope was the Hill coefficient. Curve fitting and EC50 calculations were performed using Graphpad 5.0 software.

[0343] 56.7 Results of in vitro experiments

[0344] Table 1. Electrophysiological assessment of the agonistic effect of typical compounds on the GABA _A1 receptor

[0345]

Connections 0.1 µM 1 µM KH001 D B KH002 E B KH003 D B KH004 D A KH005 D B KH006 D C KH007 E C KH008 D A KH009 D A KH010 E D KH011 D B KH012 E B KH013 E E KH014 E C KH015 E E KH016 E C KH017 E E KH018 E D KH019 E E KH020 E E KH021 E E KH022 E C KH023 E E KH024 E C KH025 E B KH026 E C KH027 E E KH028 E E KH029 E E KH030 E E KH031 E C KH032 E D KH033 E D KH034 E E KH035 E D KH036 E E KH037 E E KH038 E C KH039 E B KH040 E D KH041 E B KH042 E C KH043 E D KH045 E D KH047 E E KH048 E E KH051 E E KH052 E E Control E D

[0347] A: >80%, B: 60% - 80% (including 60%), C: 40% - 60% (including 40%), D: 20% - 40% (including 20%), E: 0 - 20%.

Table 1 shows the potency of representative compounds at concentrations of 0.1 μM and 1 μM. The potency was calculated based on the ratio of the peak current obtained at the GABA _A1 receptor to the peak current obtained with 100 μM GABA alone, and the ratio was multiplied by 100%. The peak current obtained at the GABA _A1 receptor reflected the effect of representative compounds on the GABA _A1 receptor together with 3 μM GABA at these two concentrations, respectively.

[0348] Table 2. Electrophysiological evaluation of typical compounds at the GABA _A1 receptor

[0349]

Typical connections EC ₅₀ (nmol) E _max KH002 463.9 74.6% KH003 478.1 72.8% KH004 403.7 106.1% KH022 821.1 93.1% KH025 619.9 93.7% Control 1261 84.2%

[0350] E _max was calculated as the ratio of the peak current obtained at the GABA _A1 receptor to the peak current obtained with 100 μmol GABA alone, and the ratio was multiplied by 100%. The peak current obtained at the GABA _A1 receptor reflected the effect of typical compounds on the GABA _A1 receptor together with 3 μmol GABA.

[0351] Table 3. Electrophysiological evaluation of typical compounds at the GABA _A1 receptor

[0352]

Typical connections EC ₅₀ (nmol) (without GABA) E _max
(without GABA) EC ₅₀ (nmol) (with 10 nM GABA) E _max (with 10 nM GABA) KH001 172.8 103.2% 132.2 91.7% KH004 171.6 158.4% 134.6 171.6% KH022 N/A 45.8% 385.5 88.6% KH025 947.1 84.6% 571.9 92.5% KH039 >4000 120.8% >4000 124.6% Control N/A 113.9% 218.7 148.7%

[0353] N/A indicates that the current did not change with concentration.

[0354] E _max was calculated as the ratio of the peak current obtained at the GABA _A1 receptor to the peak current obtained for GABA alone, and the ratio was multiplied by 100%. The peak current obtained at the GABA _A1 receptor reflected the effect of representative compounds alone on the GABA _A1 receptor or together with 10 nmol GABA, respectively.

[0355] Example 57. Pharmacokinetics in Brain and Plasma after Dosing

[0356] 1. Protocol

[0357] 1.1 Test medicinal products.

[0358] KH022 according to the present invention and the control described in Example 56.

[0359] 1.2 Animals

[0360] The experimental animals were CD-1 mice.

[0361] 1.3 Introduction

[0362] Test compounds were prepared with 30% sulfobutyl ether beta-cyclodextrin (SBECD) in water for intragastric administration. On the first day of the experiment, animals in the first group were administered 5 ml/kg of KH022 or control by intraperitoneal injection; animals were weighed before administration and the injection volume was calculated according to body weight. Whole blood samples (approximately 0.03 ml per group) were collected at specified times by puncture of the saphenous vein (or other sites suitable for blood collection). Blood samples were immediately processed to obtain plasma by centrifugation at approximately 4°C, 3200 g for 10 min, quickly placed on dry ice and stored at below -20°C until LC/MS/MS analysis. At each time point, animals were euthanized with CO2, and brain tissue was collected, washed with saline, dried, and homogenized with 4 times the brain weight (g) of 15 mM PBS (pH7.4):MeOH=2:1. After homogenization, samples were transferred to labeled centrifuge tubes and quickly placed on dry ice, stored at -20°C or below until LC/MS/MS analysis.

[0363] 2. Analysis of results

[0364] Test compound concentrations in input samples were determined by LC/UV and the calibration curve contained at least 6 calibration standards, ^R2 ≥ 0.999. Test compound concentrations in the biological medium were determined by LC/MS/MS. Retention times of test compounds or internal standards were calculated and chromatogram acquisition and chromatogram integration were performed using Analyst software (Applied Biosystems) and statistical processing of the results was performed in Watson LIMS (Thermo Fisher Scientific) or Analyst software (Applied Biosystems). Units of analyte concentration in the sample were ng/mL, with 3 significant digits retained, and all values expressed as percentages (e.g., % deviation and % coefficient of variation, etc.) were retained to one decimal place. Non-compartmental models of WinNonlin™ pharmacokinetic software version 6.3 (Pharsight, Mountain View, CA) were used to process plasma concentrations, and the linear log trapezoidal method was used to calculate pharmacokinetic parameters.

[0365] Experiment Results

[0366] Figure 1 and Figure 2 show that KH022 was cleared more slowly from the mouse brain than the control (control was not detected in the brain after 8 hours) at the same dosage, and the KH022 content in the mouse brain was 2.5 times higher than the control after 2 hours.

[0367] 57.3 Anxiety Absence Test in Mice

[0368] 57.3.1 Dark-Light Shuttle Chamber. The dark-light shuttle chamber test is a standard and well-characterized assay for the anxiolytic effects of representative compounds. Mice naturally avoid the light of the chamber because the light causes mice to become averse to it and experience stress, but the mice's propensity to explore also encourages them to attempt to explore the light chamber. The longer time spent in the light chamber following administration of representative compounds reflects their anxiolytic effects on mice. Experimental mice were randomly divided into 5 groups based on body weight, including a negative control group, two groups with different dosages (1 mg/kg and 3 mg/kg) of the test compounds (KH004 or KH022), and each group contained 10 male mice. Mice were treated with the appropriate dosage of the study intervention for 5 consecutive days prior to the anxiolytic experiments, which were performed 30 min after the last administration. The time spent in the light chamber was chosen as the main parameter to evaluate the anti-anxiety effects of KH004 and KH022.

[0369] Experiment Results

[0370] Table 4. Effect of KH004 and KH022 on time spent in the light chamber

[0371]

Groups Dosage
(mg/kg) Number of mice Time spent (%) Negative control group 0 10 26.69±8.38 KH022 1.0 10 50.00±4.59 ^**** 3.0 10 56.66±7.96 ^**** KH004 1.0 10 50.30±6.89 ^**** 3.0 10 53.91±9.37 ^****

[0372] Note: All results in the table are expressed as the mean ± standard deviation (mean ± SD). Statistical analysis was performed for 10 animals in each experimental group, and *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001 represent different experimental groups and the negative control animal group.

[0373] 57.3.2 Elevated Plus Maze (EPM) Testing. To analyze exploratory behavior, EPM was used to assess general locomotor and anxiety-like behavior, which consisted of two open arms and two closed arms extending from a common central platform, thus generating anxiety-like symptoms. The anxiolytic compound will relieve the conflict anxiety state of the animals in this test system. The experimental mice were randomly divided into 4 groups based on body weight, including a negative control group, two groups with different dosages (1 mg/kg and 3 mg/kg) of the test compounds (KH004 or KH022), and each group contained 10 male mice. The mice were administered the corresponding dosage of the study intervention for 5 consecutive days before the anxiolytic experiments, which were performed 30 min after the last administration. The percentage of time mice spent in the open arms was used as the primary endpoint to evaluate the anxiolytic effects of KH004 and KH022, and the results are presented in Table 5.

[0374] Table 5. Effect of different doses of KH004 and KH022 on the time mice spent in open arms

Groups Dosage
(mg/kg) Number of mice Time spent (%) Negative control group 0 10 10.79±6.24 KH022 1 10 31.40±16.41 ^** 3 10 39.46±11.82 ^**** KH004 1 10 27.21±11.15 ^*** 3 10 36.44±13.28 ^****

[0375]

[0376] Note: All results in the table are expressed as the mean ± standard deviation (mean ± SD). Statistical analysis was performed for 10 animals in each experimental group, and *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001 represent different experimental groups and the negative control animal group.

Conclusion: KH004 and KH022 significantly increased the percentage of time animals spent in the open arms, indicating their anxiolytic effect.

[0377] 57.3.3 Experiments to detect side effects during administration.

[0378] Because drugs that act on the GABA(A) receptor can cause behavioral anesthesia (side effects), side effects were observed in mice during administration in an elevated plus maze experiment. Each group of 10 mice was observed for three behavioral manifestations: gait, mobility, and proneness. "+" indicates severity, with three "+"s being the most severe, with a time to return to normal behavior of >60 minutes. The positive drug is the control compound as described in Example 56, and the results are presented in Table 6.

[0379] KH004 and KH022 had fewer side effects compared to the positive drug, as shown in Table 6.

[0380]

Table 6. Brief description of side effects observed in animals during administration

Dosage 3 mg/kg First day Second day Day three Day four Gait Mobility Being in a lying position Gait Mobility Being in a lying position Gait Mobility Being in a lying position Gait Mobility Being in a lying position Quantity Level Quantity Level Quantity Level Quantity Level Quantity Level Quantity Level Quantity Level Quantity Level Quantity Level Quantity Level Quantity Level Quantity Level Positive medicine 7 ++ 3 +++ 2 + 6 ++ 0 0 8 + 4 ++ 0 9 + 5 + 0 KH004 0 0 0 0 0 0 1 + 0 0 0 0 0 KH022 3 + 1 ++ 0 2 + 0 0 1 + 0 0 3 + 1 + 0

Claims (15)

1. A compound selected from the group consisting of: And . 2. A pharmaceutical composition having agonistic activity with respect to the GABA _A1 receptor, containing an effective amount of the compound according to claim 1 and a pharmaceutically acceptable excipient. 3. A pharmaceutical composition according to item 2, characterized in that the pharmaceutical composition is intended for oral, subcutaneous, intravenous or intramuscular administration. 4. Use of a compound according to claim 1 or a pharmaceutical composition according to claim 2 for the production of a medicinal product having agonistic activity with respect to the GABA _A1 receptor. 5. Use according to paragraph 4, characterized in that the medicinal product having agonistic activity in relation to the GABA _A1 receptor is used to regulate a neurological disease. 6. The use according to claim 5, wherein the neurological disease is selected from the group consisting of sleep disorders, mood disorders, schizophrenia spectrum disorders, spastic disorders, memory disorders and/or cognitive disorders, movement disorders, personality disorders, autism spectrum disorders, pain, traumatic brain injury, vascular diseases, disorders associated with psychoactive substance abuse, and withdrawal or tinnitus syndromes. 7. The use according to claim 6, wherein the mood disorder is depression, wherein the depression is disruptive mood disorder, major depressive disorder, persistent depressive disorder, premenstrual dysphoric disorder, substance- or drug-induced disorder, disorder due to other physical illnesses or other specified depressive disorder and non-specific depressive disorder. 8. The use according to paragraph 6, characterized in that the neurological disease is mild depression, moderate depression, severe depression or postpartum depression.