WO2025218764A1 - Compound as voltage-gated sodium channel inhibitor and use thereof - Google Patents

Abstract

The present invention relates to a voltage-gated sodium channel Nav1.8 selective inhibitor and the use thereof in the preparation of a related drug. The drug is used for treating diseases responsive to the inhibition of voltage-gated sodium channel NaV1.8, such as chronic pain, enterodynia, neuropathic pain, musculoskeletal pain, acute pain, inflammatory pain, cancer pain, idiopathic pain, post-operative pain, visceral pain, multiple sclerosis, Charcot-Marie-Tooth syndrome, incontinence, pathological cough or arrhythmia. Specifically, the present invention relates to a compound as shown in formula (X), and an isomer, nitrogen oxide, hydrate, solvate, metabolite, pharmaceutically acceptable salt or prodrug thereof.

Description

Compounds as voltage-gated sodium channel inhibitors and their uses

This application claims priority to:

CN202410483482.4, application date: April 19, 2024; CN202411125232.X, application date: August 15, 2024.

Technical Field

The present invention relates to the technical field of chemical medicine, and in particular to a compound used as a voltage-gated sodium channel inhibitor and its application.

Background Art

Pain plays an indispensable protective role in the body's normal life activities. As a warning signal, it reminds the body of potential dangers. But at the same time, pain is also a common clinical symptom. After the external stimulus that causes pain disappears, intense or persistent pain can cause physiological dysfunction and seriously affect the quality of life of the organism.

Pain originates from nociceptors in the peripheral nervous system, which are free nerve endings widely distributed in the skin, muscles, joints, and internal organs throughout the body. They can convert perceived thermal, mechanical, or chemical stimuli into nerve impulses (action potentials) and transmit them via afferent nerve fibers to their cell bodies located in the dorsal root ganglion (DRG), and ultimately to higher nerve centers, causing pain. The generation and conduction of action potentials in neurons, in turn, depend on voltage-gated sodium ion channels (NaV) on the cell membrane. When the cell membrane depolarizes, the sodium ion channels are activated, the channels open, causing an influx of sodium ions, further depolarizing the cell membrane, and leading to the generation of action potentials. Therefore, inhibiting abnormal sodium ion channel activity can help treat and relieve pain.

NaV1.8 is primarily expressed in sensory ganglia of the peripheral nervous system, such as the dorsal root ganglia (DRG), and small DRG neurons expressing NaV1.8 include pain receptors involved in pain signaling. NaV1.8 mediates large-amplitude action potentials in small neurons of the dorsal root ganglia, which are required for rapid repetitive action potentials in pain receptors and spontaneous activity of damaged neurons. Knockdown of NaV1.8 in rats has been achieved using antisense DNA or small interfering RNA and achieved almost complete reversal of neuropathic pain in spinal nerve ligation and chronic constriction injury models. Therefore, NaV1.8 channels are considered promising targets for analgesics and are expected to play a role in conditions where pain exceeds their usefulness, such as neuropathic pain, inflammatory pain, and postoperative/spontaneous pain. Moreover, since NaV 1.8 is primarily limited to neurons that sense pain, selective NaV 1.8 inhibitors may avoid the adverse events commonly seen with non-selective NaV blockers.

In addition to pain, NaV1.8 channels are also believed to be involved in diseases such as multiple sclerosis, arrhythmias, cough, and pruritus. Multiple sclerosis is an inflammatory demyelinating disease originating in the central nervous system, and its exact pathogenesis remains to be elucidated. Purkinje fibers in the cerebellum of normal people do not express NaV1.8 channels, but NaV1.8 expression is upregulated in the cerebellum of patients with multiple sclerosis. In the cardiovascular system, NaV1.8 has been shown to be expressed in cardiac nerves such as Purkinje fibers and is considered a potential therapeutic target for cardiovascular diseases such as arrhythmias. NaV1.8 is expressed in the vagal plexus associated with coughing. During pathological coughing, NaV1.8 phosphorylation levels and expression levels increase, participating in the cough reflex. In mammalian itch perception, itch factors such as histamine released by lymphocytes and mast cells can activate NaV1.8 channels. Knocking out NaV1.8 in mice can effectively alleviate histamine- and endothelin-induced itch behavior.

At present, NaV1.8 selective inhibitors, VERTEX's VX-150 and Suzetrigine have achieved positive results in clinical trials for patients with acute pain, diabetic peripheral neuropathy, etc., but there are currently no products targeting this target on the market. The development of highly selective voltage-gated sodium channel NaV 1.8 inhibitors is of great clinical significance.

Summary of the Invention

The present invention provides a compound, or a pharmaceutical composition thereof, that is a selective inhibitor of NaV1.8. The present invention further relates to the use of the compound or pharmaceutical composition thereof for preparing a medicament for treating a disease and/or condition by inhibiting NaV1.8. The present invention further describes a method for synthesizing the compound. The compound of the present invention exhibits excellent biological activity and pharmacokinetic properties.

Specifically: In one aspect, a compound is disclosed, which is a compound as shown in formula (X), or a stereoisomer, geometric isomer, tautomer, nitrogen oxide, hydrate, solvate, metabolite, pharmaceutically acceptable salt or prodrug of the compound shown in formula (X),

in:

Ring A is phenyl, 5-10 membered heteroaryl or 5-10 membered heterocyclyl;

Each R ^e is independently D, F, Cl, Br, I, CN, hydroxy, nitro, -N(R ^a ) ₂ , -(C _1-6 alkylene)N(R ^a ) ₂ , -S(O)(═NH)C _1-6 alkyl, -C(O)OR ^a , -C(O)N(R ^a ) ₂ , -(C _1-6 alkylene)C(O)N(R ^a ) ₂ , -C(O)R ^a , -S(O) ₂ NR ^a , C _1-6 alkyl, C _1-6 alkoxy, C _1-6 hydroxyalkyl, C _1-6 haloalkyl, C _1-6 haloalkoxy, C _1-6 alkylamino, C _1-6 alkylthio, C _3-6 cycloalkyl or 3-6 membered heterocyclyl, wherein the C _1-6 alkylene, C _1-6 alkyl, C 1-6 alkoxy, C _1-6 hydroxyalkyl, C _1-6 haloalkyl, C _1-6 haloalkoxy, C _1-6 alkylamino, C _1-6 alkylthio, C _3-6 cycloalkyl and 3-6 _membered heterocyclyl may be optionally substituted with 1, 2 or 3 substituents selected from D, F, Cl, Br, I, CN, hydroxy, amino, nitro, oxo, hydroxy, C _1-3 alkyl, C _1-3 alkylamino, C _1-3 haloalkyl, C _1-3 hydroxyalkyl and C _1-3 alkoxy;

each ^Ra is independently H, D, hydroxy, _C1-6 alkyl, _C1-6 alkoxy, _C3-6 cycloalkyl and 3-6 membered heterocyclyl, wherein the _C1-6 alkyl, _C1-6 alkoxy, _C3-6 cycloalkyl and _3-6 membered heterocyclyl may be optionally substituted with 1, 2 or 3 substituents selected from D, F, Cl, Br, I, CN, hydroxy, amino, nitro, oxo and _C1-3 alkyl; ^R1 , ^R2 , R3, ^R4 and ^R5 are each independently H, D, F, Cl, Br, I, CN, hydroxy, nitro, amino, C1-6 alkyl, _C1-6 alkoxy, C1-6 alkylthio, _C2-6 alkenyl, _C2-6 alkynyl, C3-6 cycloalkyl or _3-6 membered heterocyclyl, wherein the C1-6 alkyl, C1-6 alkoxy, _C3-6 cycloalkyl and _3-6 membered heterocyclyl may be optionally substituted with ¹ , 2 or 3 substituents selected from D, F, Cl, Br, I, CN, _hydroxy , amino, nitro, oxo and C1-3 _alkyl . _1-6 alkylthio, C _2-6 alkenyl, C _2-6 alkynyl, C _3-6 cycloalkyl and 3-6 membered heterocyclyl may be optionally substituted with 1, 2 or 3 substituents selected from D, F, Cl, Br, I, CN, nitro, amino, hydroxy, C _1-3 alkyl, C _1-3 haloalkyl, C _1-3 hydroxyalkyl and C _1-3 alkoxy;

or R ³ and R ⁵ together with the carbon atom to which they are attached form a C _3-6 cycloalkyl group;

R ⁶ is CN, -S(O)C _1-6 alkyl, -S(O) ₂ C _1-6 alkyl, -CH=NOC _1-6 alkyl, C _2-6 alkynyl, C _1-6 alkylthio or -L ₁ -L ₂ -R ^c , wherein the C _1-6 alkyl, C _2-6 alkynyl and C _1-6 alkylthio may be independently and optionally substituted with 1, 2 or 3 substituents selected from D, F, Cl, Br, I, oxo and hydroxy;

L ₁ is a bond, O or S;

_L2 is a bond, _C1-6 alkylene or -( _C1-6 alkylene) _-C1-6 alkoxy, wherein the _C1-6 alkylene and _C1-6 alkoxy may be independently optionally substituted with 1 or 3 substituents selected from D, F, Cl, Br, I, oxo and hydroxy;

R ^c is -ON=, -P(O)(C _1-6 alkyl) ₂ , C _2-6 alkynyl or -O-(3-8 membered heterocyclyl), wherein the C _1-6 alkyl, C _2-6 alkynyl and 3-8 membered heterocyclyl may be independently and optionally substituted with 1, 2, 3, 4, 5 or 6 substituents selected from D, F, Cl, Br, I, hydroxy, oxo, nitro, amino, alkylamino, C _1-6 alkyl, C _1-6 alkoxy, C _1-6 alkylthio, C _3-6 cycloalkyl and 3-6 membered heterocyclyl _;

^R7 and ^R8 are each independently H, D, F, Cl, Br, I, CN, hydroxyl, amino, nitro, _C1-6 alkyl, _C1-6 alkoxy, C3-6 cycloalkyl and _3-6 membered heterocyclyl, and the _C1-6 alkyl, _C1-6 alkoxy, _C3-6 cycloalkyl and 3-6 membered heterocyclyl may be optionally substituted with 1, 2 or 3 substituents selected from D, F, Cl, Br, I, CN, hydroxyl, amino, nitro and _C1-3 alkoxy.

n is 0, 1, 2 or 3.

Wherein, ring A can be phenyl, pyridazine, pyrazine, pyridine or pyrimidine.

In some embodiments, it is a compound of formula (I), (II) or (III), or a stereoisomer, geometric isomer, tautomer, nitrogen oxide, hydrate, solvate, metabolite, pharmaceutically acceptable salt or prodrug of a compound of formula (I), (II) or (III),

in:

X is CR ¹⁰ or N; Y is CR ¹³ or N;

^R9 and ^R10 are each independently H, D, F, Cl, Br, I, CN, amino, nitro, _C1-6 alkyl, _C1-6 alkoxy, _C1-6 haloalkyl or _C1-6 haloalkoxy;

R ¹¹ , R ¹² and R ¹³ are each independently H, D, F, Cl, Br, I, CN, hydroxy, nitro, -N(R ^a ) ₂ , -(C _1-6 alkylene)N(R ^a ) _{2 ,} -S(O)(═NH)C _1-6 alkyl, -C(O)OR ^a , -C(O)N(R ^a ) ₂ , -(C _1-6 alkylene)C(O)N(R ^a ) ₂ , -C(O)R ^a , -S(O) ₂ NR ^a , C _1-6 alkyl, C _1-6 alkoxy, C _1-6 hydroxyalkyl, C _1-6 haloalkyl, C _1-6 haloalkoxy, C _1-6 alkylamino, C _1-6 alkylthio, C _3-6 cycloalkyl or 3-6 membered heterocyclyl, wherein the C _1-6 alkylene, C _1-6 alkyl, C 1-6 alkoxy, C _1-6 hydroxyalkyl, C _1-6 haloalkyl, C _1-6 haloalkoxy, C _1-6 alkylamino, C _1-6 alkylthio, C _3-6 cycloalkyl and 3-6 _membered heterocyclyl may be optionally substituted with 1, 2 or 3 substituents selected from D, F, Cl, Br, I, CN, hydroxy, amino, nitro, oxo, hydroxy, C _1-3 alkyl, C _1-3 alkylamino, C _1-3 haloalkyl, C _1-3 hydroxyalkyl and C _1-3 alkoxy;

Each ^Ra is independently H, D, hydroxy, _C1-6 alkyl, _C1-6 alkoxy, _C3-6 cycloalkyl or 3-6 membered heterocyclyl, and the _C1-6 alkyl, _C1-6 alkoxy, _C3-6 cycloalkyl and 3-6 membered heterocyclyl may be optionally substituted with 1, 2 or 3 substituents selected _from D, F, Cl, Br, I, CN, hydroxy, amino, nitro, oxo and _C1-3 alkyl.

In some embodiments, wherein R ¹ , R ² , R ³ , R ⁴ and R ⁵ are each independently H, D, F, Cl, Br, I, hydroxy, methyl, ethyl, CH ₂ F, CHF ₂ , CF ₃ , -CH ₂ OCH ₃ or -CH ₂ OH.

In some embodiments, wherein R ⁹ , R ¹⁰ and R ¹¹ are each independently H, D, F, Cl, Br, I, CN, amino, nitro, methyl, ethyl, methoxy, trifluoromethyl or trifluoromethoxy;

R ¹² and R ¹³ are each independently H, D, F, Cl, Br, I, CN, hydroxyl, amino, nitro, methyl, ethyl, CH ₂ F, CHF _2, -OCH ₃ , -OCH ₂ CH ₃ , -C(O)NH ₂ , -C(O)NHOH, -C(O)NHOCH ₃ , -CH ₂ OH, -CH(OH)CH ₂ OH, -C(O)NHCH ₃ , -CH(OH)(CH ₃ ) ₂ , -S(O)(═NH)CH _{3 ,} -S(O) ₂ NH ₂ ,

In some embodiments, wherein R ⁷ and R ⁸ are each independently H, D, F, Cl, Br, I, hydroxy, methyl, ethyl, CH ₂ F, CHF ₂ or CF ₃ .

In some embodiments, wherein R ⁶ is CN, -S(O)C _1-3 alkyl, -S(O) ₂ C _1-3 alkyl-CH=NOC _1-3 alkyl, -C _2-6 alkynyl, C _1-3 alkylthio or -L ₁ -L ₂ -R ^c , said C _1-3 alkyl, C _1-3 alkylthio and C _2-6 alkynyl may be independently optionally substituted with 1, 2 or 3 substituents selected from D, F, Cl, Br, I, oxo and hydroxy;

L ₁ is a bond, O or S;

L ₂ is a bond, C _1-3 alkylene or -(C _1-3 alkylene)-C _1-3 alkoxy;

R ^c is -ON=, -P(O)(CH ₃ ) ₂ , C _2-6 alkynyl or -O-(3-8 membered heterocyclyl); the C _2-6 alkynyl and 3-8 membered heterocyclyl may be independently and optionally substituted with 1, 2, 3, 4, 5 or 6 substituents selected from D, F, Cl, Br, I, hydroxy, oxo, nitro, amino, C _1-3 alkyl, C _1-3 alkoxy, C _1-3 alkylthio, C _3-6 cycloalkyl and 3-6 membered heterocyclyl.

In some embodiments, wherein R ⁶ is CN, -S(O)CH ₃ , -S(O) ₂ CH ₃ , -P(O)(CH ₃ ) _2,

In some embodiments, the compounds disclosed herein have a structure represented by formula (IV) or (V):

wherein R ^c is a C _2-6 alkynyl group, which may be independently and optionally substituted by 1, ₂ , 3, 4, 5 or 6 substituents selected from D, F, Cl, Br, I, hydroxyl, nitro, amino, C _1-6 alkyl, C _1-6 alkoxy, C _1-6 alkylthio, C _3-6 cycloalkyl and 3-6 membered heterocyclyl.

In some embodiments, it is a compound having one of the following structures or a stereoisomer, geometric isomer, tautomer, N-oxide, hydrate, solvate, metabolite, pharmaceutically acceptable salt or prodrug having one of the following structures:

In one aspect, the present invention relates to a pharmaceutical composition comprising a compound of formula (I) of the present invention, or a stereoisomer, geometric isomer, tautomer, nitrogen oxide, hydrate, solvate, metabolite, pharmaceutically acceptable salt or prodrug thereof, and a pharmaceutically acceptable carrier, excipient, diluent, adjuvant, vehicle or a combination thereof.

In one aspect, the present invention discloses the use of the compound and pharmaceutical composition in the preparation of a medicament for treating a disease responsive to inhibition of the voltage-gated sodium channel NaV1.8, wherein the disease is chronic pain, intestinal pain, neuropathic pain, musculoskeletal pain, acute pain, inflammatory pain, cancer pain, idiopathic pain, postoperative pain, visceral pain, multiple sclerosis, Chuck-Mare-Douglas syndrome, incontinence, pathological cough, or cardiac arrhythmia.

The foregoing description only summarizes certain aspects of the present invention, but is not intended to limit the present invention to these aspects. These and other aspects will be described in more detail and fully below.

Definitions and General Terms

The present invention will list the literature corresponding to the specific content of the invention in detail, and the examples are accompanied by diagrams of structural formulas and chemical formulas. The present invention is intended to cover all options, variations and equivalents that may be included in the existing invention field as defined by the claims. Those skilled in the art will recognize many methods and materials similar or equivalent to those described herein, which can be applied to the practice of the present invention. The present invention is in no way limited to the description of methods and materials. There are many documents and similar materials that differ or conflict with the present application, including but not limited to the definition of terms, the usage of terms, the technology described, or the scope controlled by the present application.

The following definitions shall apply unless otherwise indicated. For purposes of the present invention, the chemical elements are defined according to the Periodic Table of the Elements, CAS version, and Handbook of Chemicals, 75th Ed, 1994. Additionally, general principles of organic chemistry are described in "Organic Chemistry," by Thomas Sorrell, University Science Books, Sausalito: 1999, and "March's Advanced Organic Chemistry," by Michael B. Smith and Jerry March, John Wiley & Sons, New York: 2007, all of which are hereby incorporated by reference.

The term "comprising" is an open expression, that is, including the contents specified in the present invention, but not excluding other contents.

Compounds as described herein may optionally be substituted with one or more substituents, as described in the general formulae of the present invention, or as described in the specific examples, subclasses, and classes of compounds encompassed by the present invention. It should be understood that the term "optionally substituted" is used interchangeably with the term "substituted or unsubstituted." In general, the term "optionally," whether preceded by the term "substituted," indicates that one or more hydrogen atoms in a given structure are replaced with the specified substituent. Unless otherwise indicated, an optionally substituted group may have a substituent at each substitutable position of the group. When more than one position in a given formula can be substituted with one or more substituents selected from the specified group, the substituents may be the same or different at each position. The substituents may be, but are not limited to, hydrogen, F, Cl, Br, I, nitro, cyano, oxo (=O), hydroxy, alkyl, hydroxyalkyl, alkylamino, aminoalkyl, haloalkoxy, cycloalkyl, amino, aryl, heterocyclyl, heteroaryl, alkenyl, alkynyl, cycloalkyloxy, alkoxy, alkoxyalkyl, haloalkyl, -COOH, -alkylene-C(=O)O-alkyl, -alkylene-S(=O) ₂ -alkyl, -alkylene-S(=O) ₂ -amino, -S(=O) ₂ -alkyl, -S(=O) ₂ -amino, -S(=O) _2OH , -O-alkylene-C(=O)O-alkyl, -O-alkylene-S(=O) ₂ -alkyl, -O-alkylene-S(=O) ₂ -amino, -O-alkylene-S(=O) _2OH , -C(=O) _NH2 , -C(=O)NH-alkyl, -C(=O)N(alkyl)-alkyl, -C(=O)NHS(=O) ₂ -alkyl, -C(=O)NHS(=O) ₂ -amino, -C(=O)NHS(=O) ₂ OH, -N(haloalkyl)-alkyl, -N(alkyl)-S(=O) ₂ -alkyl, -NHS(=O) ₂ -alkyl, -NHS(=O) ₂ -haloalkyl, -N(alkyl)S(=O) ₂ -haloalkyl, -N(alkyl)S(=O) ₂ -alkylamino, -NHC(=O)-alkyl, -NHC(=O)-haloalkyl, -N(alkyl)C(=O)-haloalkyl, -N(alkyl)C(=O)-alkylamino, -N(alkyl)C(=O)O-alkyl, -NHC(=O)O-alkyl, -NHC(=O)O-haloalkyl, -N(alkyl)C(=O)O-haloalkyl, -N(alkyl)C(=O)O-aminoalkyl, -NHC(=O)-NH ₂ , -NHC(=O)NH-(alkyl), -NHC(=O)NH(haloalkyl), -NHC(=O)N(alkyl)-alkyl, -OC(=O)-alkyl, -OC(=O)-amino, -OC(=O)-alkylamino, -OC(=O)-aminoalkyl, -OC(=O)-alkoxy, -C(=O)N(alkyl)S(=O) ₂ -C(═O)NH-alkyl, -C(═O)N(alkyl)-alkyl _, -C(═N-alkyl)-NH ₂ , -C(═O)NH-alkylene-S(═O) ₂ OH, -C(═O)NHC(═O)OH, -C(═O)NHC(═O)O-alkyl, -C( _═O )N(alkyl)C(═O)O-alkyl _, -C(═O)NH-alkylene-C(═O)OH, and -C(═O)NH-alkylene-C(═O)O-alkyl, and the like.

The term "alkyl" as used herein includes saturated linear or branched monovalent hydrocarbon groups of 1-20 carbon atoms, or 1-10 carbon atoms, or 1-6 carbon atoms, or 1-4 carbon atoms, or 1-3 carbon atoms, or 1-2 carbon atoms, wherein the alkyl group may be independently optionally substituted with one or more substituents described herein. Further examples of alkyl groups include, but are not limited to, methyl (Me, -CH ₃ ), ethyl (Et, -CH ₂ CH ₃ ), n-propyl (n-Pr, -CH ₂ CH ₂ CH ₃ ), isopropyl (i-Pr, -CH(CH ₃ ) ₂ ), n-butyl (n-Bu, -CH ₂ CH ₂ CH ₂ CH ₃ ), isobutyl (i-Bu, -CH ₂ CH(CH ₃ ) ₂ ), sec-butyl (s-Bu, -CH(CH ₃ )CH ₂ CH ₃ ), tert-butyl (t-Bu, -C(CH ₃ ) ₃ ), n-pentyl (-CH ₂ CH ₂ CH ₂ CH ₂ CH ₃ ), 2-pentyl (-CH(CH ₃ )CH ₂ CH ₂ CH ₃ ), 3-pentyl (-CH(CH ₂ CH ₃ ) ₂ ), 2-methyl-2-butyl (-C(CH ₃ ) ₂ CH ₂ CH ₃ ), 3-methyl-2-butyl (-CH(CH ₃ )CH(CH ₃ ) ₂ ), 3-methyl-1-butyl (-CH ₂ CH ₂ CH(CH ₃ ) ₂ ), 2-methyl-1-butyl (-CH ₂ CH(CH ₃ )CH ₂ CH ₃ ), n-hexyl (-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CH ₃ ), 2-hexyl (-CH(CH ₃ )CH ₂ CH ₂ CH ₂ CH ₃ ), 3-hexyl (-CH(CH ₂ CH ₃ )(CH ₂ CH ₂ CH ₃ )), 2-methyl-2-pentyl (-C(CH ₃ ) ₂ CH ₂ CH ₂ CH ₃ ), 3-methyl-2-pentyl (-CH(CH ₃ )CH(CH ₃ )CH ₂ CH ₃ ), 4-methyl-2-pentyl (-CH(CH ₃ )CH ₂ CH(CH ₃ ) ₂ ), 3-methyl-3-pentyl (-C(CH ₃ )(CH ₂ CH ₃ ) ₂ ), 2-methyl-3-pentyl (-CH(CH ₂ CH ₃ )CH(CH ₃ ) ₂ ), 2,3-dimethyl-2-butyl (-C(CH ₃ ) ₂ CH(CH ₃ ) ₂ ), 3,3-dimethyl-2-butyl (-CH(CH ₃ )C(CH ₃ ) ₃ ), n-heptyl, and n-octyl, among others. The term "alkyl" and its prefix "alkane" as used herein include both straight and branched saturated carbon chains.

The term "alkylene" refers to a saturated divalent hydrocarbon radical derived by removing two hydrogen atoms from a saturated straight-chain or branched hydrocarbon radical. Unless otherwise specified, an alkylene group contains 1-12 carbon atoms. In some embodiments, an alkylene group contains 1-6 carbon atoms; in other embodiments, an alkylene group contains 1-4 carbon atoms; in yet other embodiments, an alkylene group contains 1-3 carbon atoms; and in yet other embodiments, an alkylene group contains 1-2 carbon atoms. Examples include _{methylene (-CH2-), ethylene (-CH2CH2- ) ,} isopropylene (-CH( _CH3 ) _CH2- ), and the like.

The term "alkenyl" refers to a linear or branched monovalent hydrocarbon group of 2-12 carbon atoms, or 2-8 carbon atoms, or 2-6 carbon atoms, or 2-4 carbon atoms, wherein at least one position is unsaturated, i.e., one C—C is an sp ² double bond, wherein the alkenyl group may be independently and optionally substituted with one or more substituents described herein, including groups with "trans", "cis" or "E", "Z" orientations, wherein specific examples of alkenyl include, but are not limited to, vinyl (-CH=CH ₂ ), allyl (-CH ₂ CH=CH ₂ ), and the like.

The term "alkynyl" refers to a linear or branched monovalent hydrocarbon group of 2-12 carbon atoms, or 2-8 carbon atoms, or 2-6 carbon atoms, or 2-4 carbon atoms, wherein at least one position is unsaturated, i.e., one C≡C is an sp triple bond, wherein the alkynyl group may be independently and optionally substituted with one or more substituents described herein, wherein specific examples of alkynyl include, but are not limited to, ethynyl (-C≡CH), propargyl ( _-CH2C≡CH ), and the like.

The term "heteroatom" means one or more of O, S, N, P and Si, including C, N, S and P in any oxidation state; in the form of primary, secondary, tertiary amines and quaternary ammonium salts; or in the form of a nitrogen atom in a heterocyclic ring in which the hydrogen is substituted, for example, N (such as N in 3,4-dihydro-2H-pyrrolyl), NH (such as NH in pyrrolidinyl) or NR (such as NR in N-substituted pyrrolidinyl); or in the form of a _{-CH2- in a heterocyclic ring in which the -CH2-} is oxidized to form -C(=O)-.

The term "halogen" refers to F, Cl, Br or I.

The term "deuterium" refers to heavy hydrogen, D.

As used herein, the term "unsaturated" means that the moiety contains one or more degrees of unsaturation.

As used herein, the term "alkoxy" or "alkyloxy" refers to an alkyl group, as defined herein, attached to the rest of the molecule through an oxygen atom. In some embodiments, the alkoxy group is a C _1-4 alkoxy group; examples include, but are not limited to, methoxy, ethoxy, propoxy, and butoxy. The alkoxy group may be independently unsubstituted or substituted with one or more substituents as described herein.

As used herein, the term "alkylthio" or "alkylthio" refers to an alkyl group, as defined herein, attached to the remainder of the molecule through a sulfur atom. In some embodiments, the alkylthio group is a C _1-4 alkylthio group; examples include, but are not limited to, methylthio, ethylthio, propylthio, and butylthio. The alkylthio group may independently be unsubstituted or substituted with one or more substituents described herein.

As used herein, the term "alkylamino" or "alkylamino" refers to an alkyl group, as defined herein, attached to the remainder of the compound molecule via a nitrogen atom. In some embodiments, the alkylamino group is a _C1-4 alkylamino group; such examples include, but are not limited to, methylamino, ethylamino, propylamino, and butylamino. The alkylamino group may be independently unsubstituted or substituted with one or more substituents as described herein.

The term "cycloalkyl" or "cycloalkane" refers to a monovalent or polyvalent monocyclic, bicyclic, or tricyclic carbon ring system containing 3-12 carbon atoms, which may be saturated or contain one or more unsaturated bonds, but never aromatic. "Cycloalkyl" or "cycloalkane" may also include bridged and spirocyclic rings. In one embodiment, the cycloalkyl group contains 3-10 carbon atoms; in another embodiment, the cycloalkyl group contains 3-8 carbon atoms; and in yet another embodiment, the cycloalkyl group contains 3-6 carbon atoms. Examples include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cyclohexenyl. The cycloalkyl groups may independently be unsubstituted or substituted with one or more substituents described herein.

The terms "heterocyclyl" and "heterocycle" are used interchangeably herein and refer to saturated or partially unsaturated monocyclic, bicyclic, or tricyclic rings containing 3-12 ring atoms, never aromatic, in which at least one ring atom is a heteroatom. "Heterocyclyl" or "heterocycle" may also include bridged heterocycles and spiro heterocycles. In one embodiment, "heterocyclyl" or "heterocycle" contains 3-10 ring atoms; in one embodiment, "heterocyclyl" or "heterocycle" contains 3-8 ring atoms; in another embodiment, "heterocyclyl" or "heterocycle" contains 5-8 ring atoms; in yet another embodiment, "heterocyclyl" or "heterocycle" contains 3-6 ring atoms; in yet another embodiment, "heterocyclyl" or "heterocycle" contains 5-6 ring atoms; in yet another embodiment, "heterocyclyl" or "heterocycle" contains 4-6 ring atoms. Unless otherwise specified, heterocyclyl may be carbon or nitrogen radicals, and heteroatoms have the meanings described herein. Examples of heterocyclic groups include, but are not limited to, oxiranyl, azetidinyl, oxetanyl, thietanyl, pyrrolidinyl, 2-pyrrolinyl, 3-pyrrolinyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothiophenyl, dihydrothiophenyl, 1,3-dioxolane, dithiolanyl, tetrahydropyranyl, dihydropyranyl, 2H-pyranyl, 4H-pyranyl, tetrahydrothiopyranyl, piperidinyl, morpholinyl, thiomorpholinyl, piperazinyl, dioxanyl, dithianyl, thioxanyl, homopiperazinyl, homopiperidinyl, oxepanyl, thiepanyl, oxazepine Base, diazepine thiazolinone Examples of heterocyclic groups in which the -CH ₂ - group is replaced by -C(═O)- include, but are not limited to, 2-oxopyrrolidinyl, oxo-1,3-thiazolidinyl, 2-piperidinyl, 3,5-dioxopiperidinyl, pyrimidinedione, and 5,6-dihydropyridin-2(1H)-one. Examples of heterocyclic groups in which the sulfur atom is oxidized include, but are not limited to, sulfolane and 1,1-dioxothiomorpholinyl. The heterocyclic group may be optionally substituted with one or more substituents described herein.

The term "aryl" refers to monocyclic, bicyclic, and tricyclic carbocyclic ring systems containing 6-14 ring atoms, or 6-12 ring atoms, or 6-10 ring atoms, wherein at least one ring is aromatic, wherein each ring comprises 3-7 ring atoms, and has one or more points of attachment to the rest of the molecule. The term "aryl" can be used interchangeably with the term "aromatic ring". Examples of aryl groups include phenyl, naphthyl, and anthracenyl. The aryl groups may be independently optionally substituted with one or more substituents described herein.

The term "heteroaryl" refers to monocyclic, bicyclic, and tricyclic ring systems containing 5-12 ring atoms, or 5-10 ring atoms, or 5-6 ring atoms, wherein at least one ring system is aromatic and at least one ring system contains one or more heteroatoms, wherein each ring contains 5-7 ring atoms and has one or more points of attachment to the rest of the molecule. The term "heteroaryl" can be used interchangeably with the terms "heteroaromatic ring" or "heteroaromatic compound". The heteroaryl group is optionally substituted with one or more substituents described herein. In one embodiment, the 5-10 heteroaryl group contains 1, 2, 3, or 4 heteroatoms independently selected from O, S, and N, wherein the nitrogen atom can be further oxidized.

Examples of heteroaryl groups include, but are not limited to, furanyl, imidazolyl (e.g., N-imidazolyl, 2-imidazolyl, 4-imidazolyl, 5-imidazolyl), isoxazolyl, oxazolyl (e.g., 2-oxazolyl, 4-oxazolyl, 5-oxazolyl), pyrrolyl (e.g., N-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl), pyridinyl, pyrimidinyl (e.g., 2-pyrimidinyl, 4-pyrimidinyl, 5-pyrimidinyl), pyridazinyl, thiazole 1,2,3-oxadiazolyl, 1,2,5-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,3-triazolyl, 1,2,3-thiodiazolyl, 1,3,4-thiodiazolyl, 1,2,5-thiodiazolyl, 1,2,3 ... oxadiazole, pyrazinyl, 1,3,5-triazinyl; also include the following bicyclic rings, but are in no way limited to these bicyclic rings: benzimidazolyl, benzofuranyl, benzothiophenyl, indolyl (such as 2-indolyl), purinyl, quinolyl (such as 2-quinolyl, 3-quinolyl, 4-quinolyl), 1,2,3,4-tetrahydroisoquinolyl, 1,3-benzodioxolyl, indolinyl, isoquinolyl (such as 1-isoquinolyl), [1,2,4]triazolo[4,3-b]pyridazinyl, [1,2,4]triazolo[1,5-a]pyrimidinyl and [1,2,4]triazolo[1,5-a]pyridinyl, and the like.

The term "haloalkyl" or "haloalkoxy" refers to an alkyl or alkoxy group substituted with one or more halogen atoms. Examples include, but are not limited to, trifluoromethyl, trifluoromethoxy, and the like.

The term "halocycloalkyl" refers to a cycloalkyl group substituted with one or more halogen atoms. Examples include, but are not limited to, 1,1-difluorocyclopropane, 1-chloro-2-fluorocyclopropane, and the like.

As described herein, a substituent group is attached to a ring by a bond to form a ring system, which indicates that the substituent group can be substituted at any substitutable position on the ring. For example, formula (a) indicates that the substituent group R can be substituted at any substitutable position on the pyridine ring.

As described herein, a wavy line intersecting a bond in a chemical structure represents the point in the chemical structure at which the atom to which the wavy bond is attached is attached to the remainder of the molecule or to the remainder of a fragment of a molecule.

In addition, it should be noted that, unless otherwise explicitly stated, the descriptions used throughout this document, “each ... and ... are independently,” “... and ... are each independently,” and “... and ... are respectively independently,” are interchangeable and should be understood in a broad sense. They may mean that in different groups, the specific options expressed by the same symbols do not affect each other, or that in the same group, the specific options expressed by the same symbols do not affect each other.

Unless otherwise indicated, the structural formulas described herein include all isomeric forms (e.g., enantiomers, diastereomers, geometric isomers, or conformational isomers): for example, R and S configurations containing asymmetric centers, (Z) and (E) isomers of double bonds, and (Z) and (E) conformational isomers. Therefore, individual stereochemical isomers of the compounds of the present invention, or mixtures of such enantiomers, diastereomers, geometric isomers, or conformational isomers thereof, are within the scope of the present invention.

Unless otherwise indicated, the structural formulas and compounds described herein include all isomeric forms (e.g., enantiomers, diastereomers, geometric isomers, or conformers), N-oxides, hydrates, solvates, metabolites, pharmaceutically acceptable salts, and prodrugs. Therefore, individual stereochemical isomers, enantiomers, diastereomers, geometric isomers, conformers, N-oxides, hydrates, solvates, metabolites, pharmaceutically acceptable salts, and prodrugs of the compounds of the present invention are also within the scope of the present invention. Furthermore, unless otherwise indicated, the structural formulas of the compounds described herein include enriched isotopes of one or more different atoms.

"Metabolite" refers to a product obtained by metabolism in vivo of a specific compound described herein, or a pharmaceutically acceptable salt, analog, or derivative thereof, which exhibits similar activity in vivo or in vitro as the compound of formula (I). The metabolites of a compound can be identified by techniques known in the art, and their activity can be characterized by assays as described herein. Such products can be obtained by administering the compound through oxidation, reduction, hydrolysis, amidation, deamidation, esterification, defatting, or enzymatic cleavage. Accordingly, the present invention includes metabolites of a compound, including metabolites produced by contacting a compound of the present invention with a mammal for a period of time.

The definitions and conventions used in this invention are generally those of S.P. Parker, Ed., McGraw-Hill Dictionary of Chemical Terms (1984) McGraw-Hill Book Company, New York; and Eliel, E. and Wilen, S., "Stereochemistry of Organic Compounds", John Wiley & Sons, Inc., New York, 1994. The compounds of this invention may contain asymmetric centers or chiral centers and therefore exist as different stereoisomers. All stereoisomeric forms of the compounds of this invention, including but not limited to diastereomers, enantiomers, atropisomers, and mixtures thereof, such as racemic mixtures, form part of this invention. Many organic compounds exist in optically active forms, i.e., they have the ability to rotate the plane of plane-polarized light. When describing an optically active compound, the prefix D, L or R, S is used to indicate the absolute configuration of the molecule about its chiral center. The prefixes d, l, or (+), (-) are used to designate the sign of rotation of plane-polarized light in a compound. (-) or l means the compound is levorotatory, and the prefix (+) or d means the compound is dextrorotatory. These stereoisomers have the same chemical structure, but their stereostructures are different. Specific stereoisomers can be enantiomers, and a mixture of isomers is often called an enantiomeric mixture. A 50:50 mixture of enantiomers is called a racemic mixture or racemate, which may result in a lack of stereoselectivity or stereospecificity during chemical reactions. The terms "racemic mixture" and "racemate" refer to an equimolar mixture of two enantiomers that lacks optical activity.

The term "tautomer" or "tautomeric form" refers to structural isomers of different energies that are interconvertible via a low energy barrier. For example, proton tautomers (i.e., prototropic tautomers) include interconversions via proton migration, such as keto-enol and imine-enamine isomerizations. Valence tautomers include interconversions by reorganization of bonding electrons.

As used herein, "pharmaceutically acceptable salts" refer to organic and inorganic salts of the compounds of the present invention. Pharmaceutically acceptable salts are well known in the art, as described in S.M. Berge et al., describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 66:1-19, 1977. Pharmaceutically acceptable salts formed with non-toxic acids include, but are not limited to, inorganic acid salts formed by reaction with amino groups, such as hydrochlorides, hydrobromides, phosphates, sulfates, and perchlorates; organic acid salts, such as acetates, oxalates, maleates, tartrates, citrates, succinates, and malonates; or salts obtained by other methods described in the literature, such as ion exchange. Other pharmaceutically acceptable salts include adipate, malate, 2-hydroxypropionate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, cyclopentylpropionate, digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, picrate, pivalate, propionate, stearate, thiocyanate, p-toluenesulfonate, undecanoate, valerate, and the like. Salts derived from appropriate bases include alkali metal, alkaline earth metal, ammonium, and N+(C _1-4 alkyl) ₄ salts. The present invention also contemplates quaternary ammonium salts formed from any compound containing a nitrogen group. Water-soluble or oil-soluble or dispersible products can be obtained by quaternization. Alkali metals or alkaline earth metals that can form salts include sodium, lithium, potassium, calcium, magnesium, and the like. Pharmaceutically acceptable salts further include suitable, non-toxic ammonium, quaternary ammonium salts, and amine cations formed with counterions such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, C _1-8 sulfonates, and aromatic sulfonates.

The "hydrate" of the present invention refers to an association compound formed when the solvent molecule is water.

The "solvate" of the present invention refers to an association formed between one or more solvent molecules and the compound of the present invention. Solvents that form solvates include, but are not limited to, water, isopropanol, ethanol, methanol, dimethyl sulfoxide, ethyl acetate, acetic acid, and aminoethanol.

"Esters" herein refer to esters of compounds of formula (I) containing hydroxy groups that are hydrolyzable in vivo. Such esters are, for example, pharmaceutically acceptable esters that hydrolyze in the human or animal body to produce the parent alcohol. Examples of in vivo hydrolyzable esters of compounds of formula (I) containing hydroxy groups include, but are not limited to, phosphate, acetoxymethoxy, 2,2-dimethylpropionyloxymethoxy, alkanoyl, benzoyl, phenylacetyl, alkoxycarbonyl, dialkylcarbamoyl, and N-(dialkylaminoethyl)-N-alkylcarbamoyl groups.

The term "nitrogen oxide" as used herein refers to a compound containing several amine functional groups in which one or more nitrogen atoms are oxidized to form an N-oxide. Specific examples of N-oxides are N-oxides of tertiary amines or N-oxides of nitrogen atoms of nitrogen-containing heterocyclic rings. The corresponding amines can be treated with an oxidizing agent such as hydrogen peroxide or a peracid (e.g., peroxycarboxylic acid) to form N-oxides (see Advanced Organic Chemistry, Wiley Interscience, 4th edition, Jerry March, pages 1977). In particular, N-oxides can be prepared by the method of L.W. Deady (Syn. Comm. 1977, 7, 509-514), for example, by reacting the amine compound with m-chloroperbenzoic acid (MCPBA) in an inert solvent (e.g., dichloromethane).

The term "prodrug" as used herein refers to a compound that is converted in vivo to a compound represented by formula (I). Such conversion is affected by hydrolysis of the prodrug in the blood or by enzymatic conversion of the prodrug to the parent structure in the blood or tissues. The prodrug compound of the present invention may be an ester. In the prior art, esters that can be used as prodrugs include phenyl esters, aliphatic (C _1-24 ) esters, acyloxymethyl esters, carbonates, carbamates, and amino acid esters. For example, if a compound of the present invention contains a hydroxyl group, it can be acylated to obtain a prodrug form. Other prodrug forms include phosphate esters, such as these phosphate ester compounds that are obtained by phosphorylation of a hydroxyl group on the parent. For a complete discussion of prodrugs, see T. Higuchi and V. Stella, Pro-drugs as Novel Delivery Systems, Vol. 14 of the ACSSymposium Series, Edward B. Roche, ed., Bioreversible Carriers in Drug Design, American Pharmaceutical Association and Pergamon Press, 1987, J. Rautio et al, Prodrugs: Design and Clinical Applications, Nature Review Drug Discovery, 2008, 7, 255-270, and SJ Hecker et al, Prodrugs of Phosphates and Phosphonates, Journal of Medicinal Chemistry, 2008, 51, 2328-2345.

Unless otherwise indicated herein or the context clearly indicates a contrary meaning, the terms "a", "an", "the" and similar terms used in the context of the present invention (especially in the context of the claims) may be construed to include both the singular and the plural.

General synthesis process

To illustrate the present invention, the following examples are listed. However, it should be understood that the present invention is not limited to these examples, which are only provided to provide methods for practicing the present invention.

Generally, the compounds of the present invention can be prepared by the methods described herein, wherein the substituents are as defined herein unless otherwise specified. The following reaction schemes and examples are provided to further illustrate the present invention.

Those skilled in the art will recognize that the chemical reactions described herein can be used to appropriately prepare other compounds of the present invention, and that other methods for preparing the compounds of the present invention are considered to be within the scope of the present invention. For example, the synthesis of non-exemplified compounds according to the present invention can be successfully accomplished by those skilled in the art through modifications, such as appropriate protection of interfering groups, by utilizing other known reagents in addition to those described herein, or by making conventional modifications to the reaction conditions. In addition, the reactions disclosed herein or known reaction conditions are also generally applicable to the preparation of other compounds of the present invention.

Unless otherwise specified, the raw materials and reagents used in the following examples are derived from commercially available or known synthetic routes in the literature.

The equipment and testing conditions are described as follows: ^{1H NMR spectra were recorded using a Bruker 500 MHz NMR spectrometer. 1H} NMR spectra were recorded using CDCl ₃ , DMSO-d ₆ , CD ₃ OD, or acetone-d ₆ as solvents (in ppm), with TMS (0 ppm) or chloroform (7.26 ppm) as the reference standard. When multiple peaks are present, the following abbreviations are used: s (singlet), d (doublet), t (triplet), q (quartet), m (multiplet), br (broadened), brs (broadened singlet), dd (doublet of doublets), and dt (doublet of triplets). Coupling constants, J, are expressed in Hertz (Hz).

Low-resolution mass spectrometry (MS) data were measured using an Agilent G6125C quadrupole HPLC-MS (column model: XBridge BEH C18, 4.6 x 50 mm, 2.5 μm, 6 min, flow rate: 1 mL/min. Mobile phase: 0%-95% (CH ₃ CN) in (H ₂ O containing 0.1% formic acid:CH ₃ CN = 90:10), electrospray ionization (ESI), detection at 210 nm/254 nm, and DAD.

Compounds were named according to conventional nomenclature in the art or using software, and commercially available compounds were named according to the supplier's catalog.

DETAILED DESCRIPTION

The present invention is described below with reference to specific examples. It should be noted that these examples are merely illustrative and do not limit the present invention in any way.

Example 1

Preparation of compound 1-2

Under a nitrogen atmosphere, compound 1-1 (40 g, 253 mmol) was dissolved in THF (200 mL). A solution of methyllithium bromide in ether (506 mL, 759 mmol) was slowly added dropwise at zero degrees Celsius. After the addition was complete, the temperature was slowly raised to room temperature and stirred for 16 hours. After the reaction was complete, the reaction solution was cooled to 0°C and quenched by the addition of water (200 mL). After stirring for 1 hour, the solution was extracted with dichloromethane (200 mL x 5). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and distilled at 60-100°C until no more fractions were released. The remaining portion in the flask was the THF solution of compound 1-2.

¹ H NMR (400MHz, CDCl ₃ ): δ2.36 (s, 3H), 1.52 (s, 3H).

Preparation of Compounds 1-3

The preparation route of compound 1-3 is as follows:

Compound 1-4 (33.2 mL, 299.40 mmol), compound 1-5 (61.90 g, 329.34 mmol), potassium carbonate (140.68 g, 1017.96 mmol), and cuprous oxide (1.30 g, 8.98 mmol) were added sequentially to toluene (1000 mL). Tetrakistriphenylphosphine palladium (10.38 g, 8.98 mmol) was added under a nitrogen atmosphere, and the mixture was stirred at 25°C for 16 hours. The reaction solution was concentrated, water (500 mL) was added, and the mixture was extracted with ethyl acetate (500 mL × 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product, which was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 8:1) to obtain compound 1-6.

Compound 1-6 (29 g, 126 mmol) was dissolved in THF (250 mL), and a lithium hydroxide aqueous solution (144.9 mL, 289.74 mmol, 2 mol/L) was added. The reaction mixture was heated to 50°C and stirred for 1 hour. After completion of the reaction, the mixture was cooled to room temperature and washed with dichloromethane (200 mL × 1). The aqueous phase was adjusted to pH 1 with dilute hydrochloric acid (1 mol/L) and extracted with dichloromethane (100 mL × 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain compound 1-3.

Preparation of Compound 1-7

Under a nitrogen atmosphere, compound 1-3 (7.72 g, 38.2 mmol) was dissolved in acetonitrile (80 mL). Carbonyldiimidazole (6.50 g, 40.08 mmol) was added at 0°C. After stirring the mixture for 1 hour, potassium carbonate (6.59 g, 47.7 mmol) and compound 1-2 (26.6 g, 38.2 mmol, 22.4% content) were added sequentially. The reaction solution was warmed to 35°C and stirred for 10 hours. After completion of the reaction, the solid was removed by filtration, and the filtrate was concentrated under reduced pressure to obtain the crude product, which was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 10:1) to obtain compound 1-7.

¹ H NMR (400MHz, CDCl ₃ ): δ7.00-6.87 (m, 2H), 3.92 (d, J = 2.1Hz, 3H), 2.05 (s, 3H), 1.76 (s, 3H).

Preparation of Compound 1-8

Under a nitrogen atmosphere, palladium on carbon (3 g, 10% content) was added to 17.5 mL of isopropanol, and compound 1-7 (3 g, 9.31 mmol) was added with stirring at room temperature. The reaction solution was stirred at 30°C under a hydrogen atmosphere (225 psi) for 30 hours. The reaction solution was filtered through celite, and the filter cake was washed sequentially with 100 mL of isopropanol and 20 mL of dichloromethane. After concentration under reduced pressure, it was dried twice with 50 mL of toluene to obtain compound 1-8.

¹ H NMR (400MHz, CDCl ₃ ): δ7.00-6.93(m,1H),6.92-6.84(m,1H),4.49(d,J＝9.0Hz,1H),4.04(d,J ＝2.9Hz,3H),2.96-2.83(m,1H),1.72(s,3H),0.81(dd,J＝2.1,4.9Hz,3H);

m/z (ESI): [M+H] ⁺ = 325.1.

Preparation of Compound 1-9

Under a nitrogen atmosphere, diisobutylaluminum hydride (13.0 mL, 13.0 mmol, 1 mol/L toluene solution) was slowly added dropwise to a 30 mL toluene solution of compound 1-8 (2.8 g, 8.64 mmol) at -30°C. After the addition was complete, the reaction solution was stirred at -30°C for 1.5 hours. After completion of the reaction, 100 mL of ethyl acetate was added to dilute the mixture, the temperature was raised to 0°C, and saturated aqueous ammonium chloride was added dropwise to quench the mixture. The mixture was stirred for 0.5 hours and filtered through celite. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to provide compound 1-9.

¹ H NMR (400MHz, CDCl ₃ ): δ6.95-6.89(m,1H),6.89-6.80(m,1H),5.80(t,J＝3.5Hz,1H),4.00(d,J＝2.6 Hz,3H),3.88-3.78(m,1H),2.99-2.84(m,2H),1.65(s,3H),0.90-0.77(m,3H).

Preparation of Compound 1-10

Compound 1-9 (2.4 g, 7.36 mmol), 4-dimethylaminopyridine (1.35 g, 11.03 mmol), and acetic anhydride (1.0 mL, 11.0 mmol) were added sequentially to 40 mL of dichloromethane with stirring at room temperature. The reaction mixture was stirred at 20°C for 1 hour. After completion of the reaction, the mixture was slowly poured into a 30% aqueous ammonium chloride solution (100 mL) to quench the reaction. The organic phase was washed with a 10% aqueous sodium carbonate solution (100 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to yield compound 1-10.

¹ H NMR (400MHz, CDCl ₃ ): δ7.00-6.91(m,1H),6.91-6.81(m,1H),6.56(d,J＝2.3Hz,1H),4.01-3.99(m,3H),3.96(dd, J＝2.0,8.9Hz,1H),2.96-2.86(m,1H),2.11(s,3H),1.62(s,3H),0.87(dd,J＝1.9,7.6Hz,3H).

Preparation of Compound 1-11

Under a nitrogen atmosphere, compound 1-10 (2.6 g, 7.06 mmol) was added to 40 mL of dichloromethane. After cooling to -30°C, trimethylsilyl cyanide (2.8 mL, 21.2 mmol) was slowly added to the reaction solution. After stirring for 5 minutes, boron trifluoride etherate (7.8 mL, 28.2 mmol) was added dropwise. The reaction solution was stirred at -30-0°C for 2 hours. After completion of the reaction, potassium hydroxide aqueous solution (2 mol/L, 100 mL) was slowly added dropwise. The mixture was extracted with dichloromethane (50 mL x 2). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain compound 1-11.

¹ H NMR (400MHz, CDCl ₃ ): δ6.93-6.85(m,1H),6.78-6.76(m,1H),5.08-5.01(m,1H),4.26-4.21(m,1H),4.08-4.05(m,3H),2.89 -2.81(m,1H),1.64(s,3H),0.83-0.79(m,3H).

Preparation of Compound 1-12

Compound 1-11 (2.4 g, 7.16 mmol) was added to 20 mL of ethanol, and a 20 mL, 2 mol/L potassium hydroxide aqueous solution was added dropwise with stirring. The reaction mixture was heated to 75°C and stirred for 16 hours. After completion of the reaction, the mixture was cooled and diluted with 100 mL of water. The ethanol was then concentrated under reduced pressure to remove the ethanol. 50 mL of ethyl acetate was added to the concentrate, and the aqueous phase was adjusted to pH 5 with 6 M hydrochloric acid. The mixture was extracted with ethyl acetate (50 mL x 2). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by reverse-phase column chromatography (mobile phase A: water (0.1% formic acid) - B: acetonitrile; elution gradient: B: 70%-80%; elution over 15 minutes) to obtain compound 1-12.

¹ H NMR (400MHz, DMSO-d ₆ )δ13.13-12.80(m,1H),7.22-7.16(m,1H),7.16-7.09(m,1H),4.98(d,J＝10.4Hz,1H),4.08(dd ,J＝7.9,10.8Hz,1H),3.93(d,J＝2.1Hz,3H),2.73-2.60(m,1H),1.53(s,3H),0.74-0.64(m,3H).

m/z(ESI):[MH] ^- ＝353.2.

Preparation of Compound 1-13

Compound 1-12 (2.1 g, 5.93 mmol) was added to 4 mL of toluene, stirred and heated to 60 ° C, and then R-phenylethylamine (0.86 g, 7.11 mmol) in 4 mL of toluene was slowly added dropwise. After the addition was complete, the reaction temperature was lowered to 50 ° C and stirred for 1 hour. The reaction temperature was slowly cooled to 20 ° C and stirred at 20 ° C for 8 hours. The mixture was filtered, the filter cake was washed with 5 mL of toluene, and dried to obtain compound 1-13.

¹ H NMR (400MHz, MeOD): δ7.50-7.41(m,5H),7.13(ddd,J＝2.1,6.0,8.6Hz,1H),6.96-6.87(m,1H),4.73(d,J＝10.4Hz,1H),4.48 -4.38(m,1H),4.09(dd,J＝8.3,10.1Hz,1H),3.96(d,J＝2.1Hz,3H),2.58-2.48(m,1H),1.65-1.57(m,6H),0.78-0.70(m,3H).

Preparation of Compound 1-14

Compound 1-13 (2.25 g, 4.73 mmol) was added to a mixed solution of 20 mL of isopropanol and 40 mL of n-heptane, and 40 mL of dichloromethane and 40 mL of aqueous hydrochloric acid (2-mol/L) were added in sequence with stirring. The reaction solution was stirred for half an hour and concentrated under reduced pressure to remove the low-boiling solvent. 50 mL of ethyl acetate was added to the residual liquid, and the liquid was extracted. The organic phase was washed with 20 mL of water, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain an oil (1.76 g). Quinine (1.56 g, 4.80 mmol) was added to a 4 mL dichloromethane solution containing the above oil. 4 mL of isopropanol was added and the temperature was raised to 70°C (low-boiling dichloromethane was evaporated). After the reaction solution temperature dropped to 65°C, 12 mL of n-heptane was slowly added. After stirring at 65°C for 1 hour, the reaction solution was slowly cooled to 20°C. After stirring at 20°C for 2 hours, the solution was filtered and the filter cake was washed with isopropanol/n-heptane = 1/3 (3 mL) and dried under reduced pressure to obtain compound 1-14.

¹ H NMR (400MHz, CDCl ₃ )δ8.68(d,J＝4.5Hz,1H),7.84(d,J＝9.1Hz,1H),7.60(d,J＝4.4Hz,1H),7.18(dd,J＝2.6,9.3Hz,1H),7.10(ddd,J＝1.8,6.1,8.4Hz, 1H),6.84-6.75(m,2H),6.16(s,1H),5.52(ddd,J＝6.7,10.4,17.2Hz,1H),5.06-4.92(m,2H),4.85(d,J＝10.1Hz,1H),4.37-4.23(m ,1H),4.17-4.07(m,1H),3.93(d,J＝2.5Hz,3H),3.70(s,3H),3.39-3.24(m,2H),3.07-2.91(m,2H),2.60-2.48(m,1H),2.58(d,J＝ 1.5Hz,1H),2.15-2.04(m,1H),2.03-1.93(m,2H),1.80-1.68(m,1H),1.65(s,3H),1.27-1.10(m,1H),0.78(dd,J＝1.8,7.3Hz,3H).

Preparation of Compound 1-15

20 mL of dilute hydrochloric acid (2.5 mol/L) was added dropwise to a solution of compound 1-14 (1.6 g, 2.36 mmol) in 20 mL of dichloromethane. The mixture was stirred at 20°C for half an hour and then allowed to stand for separation. 30 mL of dilute hydrochloric acid (2.5 mol/L) was added to the organic phase again and the phases separated. The combined aqueous phases were extracted with dichloromethane (20 mL × 2). The combined organic phases were washed with 20 mL of water, dried over anhydrous sodium sulfate, and concentrated to obtain a white solid (600 mg). Under a nitrogen atmosphere, DMF (0.1 mL, 0.34 mmol) and oxalyl chloride (0.3 mL, 3.39 mmol) were added dropwise in sequence to a solution of the above white solid (600 mg, 1.69 mmol) in 10 mL of dichloromethane at zero degrees Celsius. The reaction solution was warmed to room temperature, stirred for 2 hours, and concentrated under reduced pressure to obtain compound 1-15.

Preparation of Compound 1-16

Under a nitrogen atmosphere, compound 1-15 (200 mg, 0.54 mmol) was slowly added to methyl 4-amino-2-picolinate (163.3 mg, 1.07 mmol) and triethylamine (0.2 mL, 1.61 mmol) in 4 mL of dichloromethane at zero degrees Celsius. The mixture was warmed to 20°C and stirred for 2 hours. After completion of the reaction, 20 mL of water was added dropwise to quench the reaction. The mixture was extracted with dichloromethane (10 mL x 2). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product, which was then purified by reverse-phase column chromatography (mobile phase A: water (0.1% ammonia) - B: acetonitrile; elution gradient: B: 70%-80%; elution over 15 minutes) to obtain compound 1-16.

¹ H NMR (400MHz, CDCl ₃ ): δ8.64(d,J＝5.4Hz,1H),8.58(s,1H),8.09(d,J＝2.1Hz,1H),7.94(dd,J＝2.3,5.5Hz,1H),7.12-7.05(m,1H),6.97-6. 88(m,1H),5.04(d,J=11.1Hz,1H),4.21-4.06(m,2H),2.84-2.70(m,1H),1.70(s,3H),1.56(s,4H),0.86-0.75(m,3H);

m/z (ESI): [M+H] ⁺ = 489.2.

Preparation of Compound 1-17

Compound 1-16 (210 mg, 0.43 mmol) was added to a 7 mol/L ammonia/methanol solution (6.1 mL, 43.00 mmol), and the reaction mixture was incubated at 40°C in a tetrafluoroethylene flask for 24 hours. After completion of the reaction, the crude product was concentrated and purified by preparative HPLC (column model: Phenomenex luna C18 150*25 mm*10 μm; mobile phase: [A: water (0.1% formic acid)-B: acetonitrile; elution gradient: B%: 40%-70% over 10 minutes) to obtain compound 1-17.

¹ H NMR (400MHz, DMSO-d ₆ ): δ10.74 (s, 1H), 8.49 (d, J = 5.5Hz, 1H), 8.28 (d, J = 2.0Hz, 1H), 8.07 (br d,J＝2.1Hz,1H),7.83(dd,J＝2.1,5.5Hz,1H),7.63(s,1H),7.22-7.12(m,2H),5.10(d,J＝10.3Hz,1H),4. 25(dd,J＝7.9,10.1Hz,1H),3.94(d,J＝2.0Hz,3H),2.83-2.71(m,1H),1.61(s,3H),0.73(d,J＝6.5Hz,3H);

m/z (ESI): [M+H] ⁺ = 474.4.

Preparation of Compound 1-18

Under a nitrogen atmosphere, boron tribromide (0.30 mL, 0.32 mmol) was slowly added dropwise to a solution of compound 1-17 (100 mg, 0.21 mmol) in 2 mL of dichloromethane at zero degrees Celsius. After the addition was complete, the mixture was stirred at zero degrees Celsius for 2 hours. After completion of the reaction, the reaction solution was poured into 50 mL of saturated aqueous sodium bicarbonate solution to quench the reaction. The aqueous phase was extracted with ethyl acetate (20 mL x 2). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product, which was then purified on thin-layer silica gel (dichloromethane/methanol = 10:1) to obtain compound 1-18.

¹ H NMR (400MHz, DMSO-d ₆ ): δ10.74(s,1H),10.53-10.41(m,1H),8.49(d,J＝5.5Hz,1H),8.27(s,1H),8.06(s,1H),7.83(dd,J＝2.1,5.5Hz,1H),7.62(br s,1H),7.03(t,J＝7.1Hz,1H),6.86(q,J＝8.9Hz,1H),5.10(d,J＝10.3Hz,1H),4.30-4.20(m,1H),2.90-2.78(m 1H),2.54(d,J＝1.0Hz,2H),1.60(s,3H),0.71(d,J＝6.3Hz,3H);

m/z(ESI):[M+H] ⁺ =460.2.

Preparation of Example 1

Under nitrogen atmosphere, propargyl bromide (2.38 mg, 0.02 mmol) was slowly added dropwise to a 1 mL DMF solution of compound 1-18 (10 mg, 0.02 mmol) and potassium carbonate (13.8 mg, 0.02 mmol). After the addition was complete, the temperature was raised to 60°C and the reaction was allowed to proceed for 2 hours. The reaction solution was cooled and filtered, and the filtrate was purified by preparative high performance liquid separation (column model: Waters Xbridge 150*25mm*5μm; mobile phase A: water (ammonia water)-B: acetonitrile; elution gradient: B%: 42%-72%; elution time: 11 minutes) to obtain Example 1.

¹ H NMR (400 MHz, CDCl ₃ ): δ8.68(s,1H),8.46(d,J＝5.6Hz,1H),8.16(dd,J＝2.2,5.6Hz,1H),7.94(d, J＝2.1Hz,1H),7.84(s,1H),7.19-7.11(m,1H),7.05-6.94(m,1H),5.67-5.56( m,1H),5.02(d,J＝11.4Hz,1H),4.93-4.74(m,2H),4.26(dd,J＝7.8,11.3Hz,1 H),2.90-2.79(m,1H),2.38(t,J＝2.4Hz,1H),1.72(s,3H),0.86-0.77(m,3H);

m/z (ESI): [M+H] ⁺ = 498.2.

Example 2

Under nitrogen atmosphere, 1-bromo-2-butyne (14.5 mg, 0.11 mmol) was slowly added dropwise to a 1 mL DMF solution containing compound 1-18 (50 mg, 0.11 mmol) and potassium carbonate (75.2 mg, 0.54 mmol). The reaction solution was heated to 60°C for 2 hours. After the reaction was completed, the reaction solution was cooled to room temperature and filtered. The filtrate was purified by preparative HPLC separation (column model: Waters Xbridge 150*25 mm*5 μm; mobile phase A: water (ammonium bicarbonate)-B: acetonitrile; elution gradient: B%: 50%-80% elution over 9 minutes) to obtain Example 2.

¹ H NMR (400 MHz, DMSO-d ₆ )δ10.72(s,1H),8.49(d,J＝5.6Hz,1H),8.28(d,J＝2.0Hz,1H),8.10-8.02(m ,1H),7.84(dd,J＝2.0,5.6Hz,1H),7.62(d,J＝2.0Hz,1H),7.28-7.14(m,2H) ,5.12(d,J＝10.4Hz,1H),4.93-4.78(m,2H),4.37(dd,J＝7.6,10.3Hz,1H),2 .90-2.70(m,1H),1.75(t,J＝2.4Hz,3H),1.62(s,3H),0.73(d,J＝6.4Hz,3H);

m/z (ESI): [M+H] ⁺ = 512.2.

Example 3

Preparation of compound 3-1

Under a nitrogen atmosphere, compound 1-18 (200 mg, 0.04 mmol) and triethylamine (0.1 mL, 0.65 mmol) were dissolved in 3 mL of dichloromethane. The mixture was stirred at 0°C for 10 minutes. Trifluoromethanesulfonic anhydride (72.3 μL, 0.44 mmol) was added dropwise. After the addition was complete, the reaction was continued at 0°C for 1 hour. After completion of the reaction, the reaction solution was concentrated under reduced pressure to obtain a crude product, which was then isolated and purified by silica gel column chromatography (petroleum ether:ethyl acetate = 1:1) to obtain compound 3-1.

m/z(ESI):[M+H] ⁺ =592.1.

Preparation of compound 3-2

Under nitrogen atmosphere, 1,1'-bis(diphenylphosphino)ferrocenepalladium(II) chloride (6.19 mg, 0.01 mmol), cuprous iodide (3.22 mg, 0.02 mmol), and triethylamine (25.66 mg, 0.25 mmol) were added sequentially to 0.5 mL of DMF solvent containing compound 3-1 (50 mg, 0.08 mmol) and triisopropylsilyl acetylene (0.1 mL, 0.25 mmol). The reaction mixture was heated to 100°C and stirred for 6 hours. After the reaction was completed, the reaction mixture was cooled to room temperature and filtered through a short silica gel (washed with ethyl acetate). The filtrate was concentrated under reduced pressure to obtain the crude product, which was purified by reverse phase column chromatography (0.1% formic acid system) to obtain compound 3-2.

¹ H NMR (400 MHz, CDCl ₃ ): δ8.61(s,1H),8.47(d,J＝5.6Hz,1H),8.18(dd,J＝2.1,5.4Hz,1H),7.89(d,J＝ 2.0Hz,1H),7.86(d,J＝4.5Hz,1H),7.18(d,J＝5.7Hz,2H),5.59(d,J＝2.9Hz,1H), 5.03(d,J＝11.2Hz,1H),4.32(dd,J＝8.2,11.0Hz,2H),2.97(t,J＝7.6Hz,1H),2. 37(s,1H),2.03(d,J=6.6Hz,1H),1.68(s,3H),1.10(s,18H),0.87-0.81(m,3H);

m/z (ESI): [M+H] ⁺ = 624.2.

Preparation of Example 3

Under nitrogen atmosphere, cesium fluoride (30.38 mg, 0.20 mmol) was added to a solution of compound 3-2 (13 mg, 0.02 mmol) in 0.5 mL of DMF, and the reaction mixture was stirred at 25°C for 1 hour. After completion of the reaction, the reaction mixture was filtered, and the filtrate was purified by preparative HPLC (Waters Xbridge 150*25 mm*5 μm; mobile phase A: water (ammonium bicarbonate)-B: acetonitrile; elution gradient: B%: 45%-75%; elution over 9 minutes) to obtain Example 3.

¹ H NMR (400MHz, CDCl ₃ ): δ8.67(s,1H),8.48(d,J＝5.5Hz,1H),8.17(dd,J＝2.0,5.4Hz,1H),7.94(d,J＝1.6Hz,1H),7.87(s,1H),7.26-7.17(m,2H),5.61(d,J＝ 2.3Hz,1H),5.06(d,J＝11.0Hz,1H),4.27(dd,J＝7.9,11.2Hz,1H),3.64(s,1H),2.97(t,J＝7.6Hz,1H),1.71(s,3H),0.88-0.74(m,3H);

m/z (ESI): [M+H] ⁺ = 468.2.

Example 4

Preparation of compound 4-2

The synthetic route of compound 4-2 is as follows:

Under nitrogen atmosphere, compound 4-1 (3.00 g, 18.4 mmol) was added dropwise to a 50 mL DMF solution containing 1,2-dibromoethane (14.5 g, 77.2 mmol) and triethylamine (5.60 mL, 40.5 mmol). The mixture was stirred at 20°C for 16 hours. After the reaction was completed, the reaction solution was poured into 100 mL of water and the mixture was extracted with ethyl acetate (50 mL × 2). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product, which was then separated and purified by silica gel column chromatography (eluted with petroleum ether) to obtain compound 4-2.

¹ H NMR (400MHz, DMSO-d ₆ ): δ7.87 (s, 4H), 4.45 (t, J = 6.0Hz, 2H), 3.77-3.77 (m, 1H), 3.75 (t, J = 6.0Hz, 1H).

Preparation of compound 4-3

Under nitrogen, compound 4-2 (58.8 mg, 0.22 mmol) was added to a 2 mL DMF solution of compound 1-18 (100 mg, 0.22 mmol) and potassium carbonate (150 mg, 1.09 mmol). The reaction mixture was heated to 60°C and stirred for 6.5 hours. After the reaction was complete, the reaction mixture was cooled to room temperature, filtered, and the filtrate was purified by reverse-phase column chromatography (0.1% formic acid) to obtain compound 4-3.

¹ H NMR (400 MHz, CDCl ₃ ): δ8.75(s,1H),8.35(d,J＝5.6Hz,1H),8.23(dd,J＝2.4,5.6Hz,1H),7.80(d,J ＝2.0Hz,2H),7.70(s,4H),7.25-7.17(m,1H),7.02-6.92(m,1H),5.82-5.71(m, 1H),5.00(d,J＝11.6Hz,1H),4.74-4.62(m,2H),4.51(dd,J＝8.0,10.4Hz,1H),4 .44-4.25(m,2H),3.07-2.97(m,1H),1.78(s,3H),0.80(dd,J＝2.0,5.6Hz,3H);

m/z (ESI): [M+H] ⁺ = 649.2.

Preparation of compound 4-4

Under nitrogen atmosphere, hydrazine hydrate (14.2 mg, 0.28 mmol) was added to a solution of compound 4-3 (90.0 mg, 0.14 mmol) in 3 mL of ethanol. The reaction mixture was heated to 65°C and stirred for 1 hour. After the reaction was complete, the reaction mixture was cooled to room temperature, filtered, and the filtrate was concentrated to obtain compound 4-4.

¹ H NMR (400MHz, CDCl ₃ ): δ8.70-8.64(m,1H),8.47(d,J＝5.6Hz,1H),8.13(dd,J＝2.4,5.6Hz,1H),7.95(d,J ＝2.0Hz,1H),7.87-7.80(m,1H),7.13-7.06(m,1H),6.99(d,J＝2.4Hz,1H),5.62-5.5 6(m,1H),5.02(d,J＝11.2Hz,1H),4.49-4.40(m,1H),4.38-4.23(m,2H),3.96-3.88( m,1H),3.86-3.79(m,1H),2.88-2.74(m,1H),1.29-1.23(m,3H),0.83-0.77(m,3H).

Preparation of Example 4

Under a nitrogen atmosphere, pyridine p-toluenesulfonate (0.27 mg) and paraformaldehyde (6.37 mg, 0.21 mmol) were added sequentially to a 3 mL dichloromethane solution of compound 4-4 (55 mg, 0.11 mmol), and the reaction solution was heated to 40 ° C and stirred for 4 hours. After the reaction was completed, water (20 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (10 mL × 2). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product, which was purified by preparative high performance liquid phase separation (column model: Waters Xbridge 150*25mm*5um; mobile phase A: water (ammonium bicarbonate)-B: acetonitrile; gradient: B: 48%-78% elution for 10 minutes) to obtain Example 4.

¹ H NMR (400MHz, DMSO-d ₆ ): δ10.69(s,1H),8.49(d,J＝5.6Hz,1H),8.28(d,J＝2.0Hz,1H),8.07(d,J＝2.0Hz, 1H),7.84(dd,J＝2.4,5.6Hz,1H),7.62(d,J＝2.0Hz,1H),7.20-7.13(m,2H),7.05(d ,J＝7.6Hz,1H),6.58(d,J＝7.6Hz,1H),5.12(d,J＝10.6Hz,1H),4.43(dd,J＝4.8,6.8 Hz,1H),4.37-4.23(m,4H),2.88-2.76(m,1H),1.60(s,3H),0.70(d,J＝6.0Hz,3H);

m/z (ESI): [M+H] ⁺ = 531.2.

Example 5

Preparation of compound 5-2

The synthetic route of compound 5-2 is as follows:

Under nitrogen atmosphere, tert-butyldimethylsilyl chloride (865 mg, 5.74 mmol) was added to a 5 mL DMF solution of compound 5-1 (300 mg, 2.87 mmol) and imidazole (586 mg, 8.61 mmol), and the reaction solution was stirred at 20°C for 3 hours. After the reaction was completed, water (50 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (25 mL × 2). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product, which was separated and purified by silica gel column chromatography (eluted with petroleum ether) to obtain compound 5-2.

¹ H NMR (400MHz, CDCl ₃ ): δ4.37 (t, J = 2.0 Hz, 2H), 4.18 (t, J = 2.0 Hz, 2H), 0.92 (s, 9H), 0.13 (s, 6H).

Preparation of compound 5-3

Under nitrogen atmosphere, a DMF solution (1 mL) of compound 5-2 (28.6 mg, 0.13 mmol) was slowly added to a suspension of compound 1-18 (60 mg, 0.13 mmol) and potassium carbonate (90.3 mg, 0.65 mmol) in DMF (1 mL). The reaction mixture was heated to 60°C and stirred for 2 hours. After completion of the reaction, the reaction mixture was cooled to room temperature and filtered. The filtrate was isolated and purified using reverse phase preparative chromatography (mobile phase A: water (0.1% ammonia) - B: acetonitrile; elution gradient: B: 100%; elution over 3 minutes) to obtain compound 5-3.

¹ H NMR (400 MHz, DMSO-d ₆ ): δ10.73(s,1H),8.48(d,J＝5.6Hz,1H),8.27(d,J＝2.0Hz,1H),8.05(d,J＝2 .4Hz,1H),7.95(s,4H),7.84(dd,J＝2.4,5.6Hz,1H),7.61(d,J＝2.4Hz,1H), 7.26-7.17(m,2H),5.12(d,J＝10.4Hz,1H),5.04-4.92(m,2H),4.33(dd,J＝7 .6,10.2Hz,1H),4.28-4.22(m,2H),1.61(s,3H),0.77(s,9H),-0.06(s,6H);

m/z (ESI): [M+H] ⁺ = 642.3.

Preparation of Example 5

Compound 5-3 (25 mg, 0.04 mmol) was dissolved in a methanol solution of hydrogen chloride (2 mL, 2 mol/L) and stirred at 25°C for 0.5 h. After completion of the reaction, the mixture was concentrated, diluted with 2 mL of methanol, and adjusted to pH 8 with ammonia water. The mixture was purified by preparative HPLC (column model: Waters Xbridge 150*25 mm*5 μm; mobile phase A: water (ammonium bicarbonate)-B: acetonitrile; elution gradient: B: 38%-68%; elution time: 9 min) to obtain Example 5.

¹ H NMR (400MHz, DMSO-d ₆ ): δ10.73 (s, 1H), 8.49 (d, J = 5.6 Hz, 1H), 8.26 (d, J = 2.0 Hz, 1H), 8.08 (d, J = 2.0 Hz, 1H), 7.84 (dd, J = 2.0, 5.6 Hz, 1H), 7.63 (br d,J＝1.6Hz,1H),7.28-7.14(m,2H),5.21(t,J＝6.0Hz,1H),5.11(d,J＝10.4Hz,1H),4.94(d,J＝1.6Hz,2H) ,4.34(dd,J＝7.6,10.4Hz,1H),4.10-4.03(m,2H),2.90-2.79(m,1H),1.62(s,3H),0.74(d,J＝6.4Hz,3H);

m/z (ESI): [M+H] ⁺ = 528.2.

Example 6

Preparation of compound 6-2

Under a nitrogen atmosphere, methyllithium (5.3 mL, 8.41 mmol) was slowly added dropwise to compound 6-1 (1.0 g, 8.41 mmol) in 15 mL of tetrahydrofuran at -78°C. The reaction mixture was stirred at -78°C for 5 minutes, followed by the slow addition of acetone (6.2 mL, 84.06 mmol). After the addition was complete, the reaction mixture was stirred at -78°C for 10 minutes. After completion of the reaction, 30 mL of saturated aqueous ammonium chloride was added to quench the reaction. The aqueous phase was extracted with ethyl acetate (30 mL x 3). The combined organic phases were washed with 50 mL of saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product, which was then purified by silica gel column chromatography (petroleum ether:ethyl acetate = 10:1) to obtain compound 6-2.

¹ H NMR (400MHz, CDCl ₃ ): δ3.94 (s, 2H), 1.53 (s, 6H).

Preparation of Example 6

Under nitrogen atmosphere, potassium carbonate (63.18 mg, 0.46 mmol) and compound 6-2 (16.29 mg, 0.09 mmol) were added to a 1 mL DMF solution of compound 1-18 (50 mg, 0.09 mmol), and the reaction solution was stirred at 60°C for 2 hours. After the reaction was completed, the solution was filtered, and the filtrate was purified by preparative HPLC (column model: Waters Xbridge 150*25 mm*5 μm; mobile phase A: water (ammonium bicarbonate)-B: acetonitrile; elution gradient: B: 35% to 65%; elution time 10 minutes) to obtain Example 6.

¹ H NMR (400 MHz, DMSO-d ₆ ): δ10.74(s,1H),8.49(d,J＝5.6Hz,1H),8.27(d,J＝2.0Hz,1H),8.06(d,J＝2.0Hz ,1H),7.84(dd,J＝2.0,5.6Hz,1H),7.61(d,J＝1.6Hz,1H),7.24-7.15(m,2H),5.3 1(s,1H),5.12(d,J＝10.6Hz,1H),4.94(s,2H),4.28(dd,J＝7.2,10.2Hz,1H),2.8 5(t,J＝7.2Hz,1H),1.63(s,3H),1.27(s,3H),1.24(s,3H),0.74(d,J＝5.6Hz,3H);

m/z (ESI): [M+H] ⁺ = 556.3.

Example 7

Under a nitrogen atmosphere, 2-methyl-3-butyn-2-ol (0.1 mL, 0.89 mmol) and _K₂CO₃ ( _451.27 mg, 3.27 mmol) were sequentially added to a solution of compound 1-18 (300 mg, 0.65 mmol) in DMF (6 mL). The reaction solution was reacted at 100°C for 2 hours. Compound 2 (0.1 mL, 0.89 mmol) was added to the reaction solution, and the reaction solution was continued at 100°C for 2 hours. After completion of the reaction, the reaction solution was cooled to room temperature and filtered. The filtrate was purified by reverse phase preparative purification (Waters Xbridge 150×25 mm× ₅ μm, water (water ( _NH₄HCO₃ )-ACN, 52%-82%, 16 min) to obtain Example 7.

¹ H NMR (400MHz, DMSO-d ₆ ): δ10.76 (br s,1H),8.48(d,J＝5.52Hz,1H),8.25(d,J＝1.88Hz,1H),8.08(d,J＝2.00Hz,1H),7.82(dd,J＝5.40,1.88Hz,1H),7.63(br s,1H),7.16-7.29(m,2H),5.09(d,J＝10.52Hz,1H),4.43(dd,J＝10.32,7.60Hz,1H), 3.54(s,1H),2.80(t,J＝7.20Hz,1H),1.70(s,3H),1.67(s,3H),1.61(s,3H),0.70(br d, J = 5.12 Hz, 3H);

m/z(ESI):[M+H] ⁺ =526.1.

Example 8

Preparation of compound 8-2

Under a nitrogen atmosphere, methanesulfonic anhydride (0.80 mL, 6.42 mmol) was slowly added to a solution of compound 8-1 (300 mg, 4.28 mmol) and triethylamine (1.8 mL, 12.8 mmol) in dichloromethane (5 mL). The reaction mixture was stirred at 20°C for 2 hours. After completion of the reaction, the reaction mixture was concentrated under reduced pressure, diluted with water (50 mL), and extracted with ethyl acetate (25 mL x 2). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain compound 8-2.

¹ H NMR (400MHz, CDCl ₃ ): δ5.30 (dq, J=2.0, 6.8Hz, 1H), 3.13 (s, 3H), 2.71 (d, J=2.0Hz, 1H), 1.67 (d, J=6.8Hz, 3H).

Preparation of Example 8

Under nitrogen atmosphere, a solution of compound 8-2 (6.45 mg, 0.04 mmol) in DMF (0.5 mL) was slowly added to a solution of compound 1-18 (100 mg, 0.22 mmol) and potassium carbonate (150 mg, 1.09 mmol) in DMF (0.5 mL). The reaction solution was stirred at 60°C for 16 hours. After completion of the reaction, the reaction solution was filtered, and the filtrate was purified by reverse phase preparative purification (column: Waters Xbridge 150×25 mm×5 μm, water (NH ₄ HCO ₃ )-ACN, 40%-70%, 15 min) to obtain Example 8.

¹ H NMR (400MHz, DMSO-d ₆ ): δ10.70 (s, 1H), 8.48 (d, J = 5.6Hz, 1H), 8.25 (d, J = 2.0Hz, 1H), 8.07 (d, J = 2.0Hz, 1H), 7.82 (dd, J = 2.0, 5.6Hz, 1H), 7.62 (br d,J＝2.0Hz,1H),7.27-7.14(m,2H),5.25(dq,J＝2.0,6.4Hz,1H),5.10(d,J＝10.8Hz,1H),4.31 (dd,J＝7.6,10.8Hz,1H),3.67(d,J＝2.0Hz,1H),2.85-2.73(m,1H),1.66-1.57(m,6H),0.77(br d,J=6.0Hz,3H);

m/z (ESI): [M+H] ⁺ = 512.2.

Example 9

Under a nitrogen atmosphere, cyanomethylenetri-n-butylphosphine (132.4 mg, 0.55 mmol) and (S)-(-)-3-butyn-2-ol (38.45 mg, 0.55 mmol) were added to a solution of compound 1-18 (60 mg, 0.11 mmol) in 1,4-dioxane (2 mL). The reaction was stirred at 40°C for 16 hours. After completion, the reaction was quenched with water (40 mL) and extracted with ethyl acetate (30 mL × 2). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product, which was then purified by reverse phase preparative purification (Column: Waters Xbridge 150×25 mm×5 μm; Condition: water (NH ₄ HCO ₃ )-ACN; Begin B: 43, End B: 73; Gradient Time (min): 10; 100% B Hold Time (min): 4, Flow Rate (ml/min): 25) to obtain Example 9.

¹ H NMR (400MHz, DMSO-d ₆ ): δ10.72 (s, 1H), 8.49 (d, J = 5.6 Hz, 1H), 8.26 (d, J = 2.0 Hz, 1H), 8.07 (d, J = 2.0 Hz, 1H), 7.84 (dd, J = 2.0, 5.6 Hz, 1H), 7.63 (br s,1H),7.30-7.15(m,2H),5.16-5.06(m,2H),4.45(dd,J＝7.2,10.2Hz,1H),3.4 1(d,J＝2.0Hz,1H),2.91-2.79(m,1H),1.66-1.59(m,6H),0.70(d,J＝6.0Hz,3H);

m/z(ESI):[M+H] ⁺ =512.1.

Example 10

Preparation of compound 10-2

Under a nitrogen atmosphere, tert-butyldimethylsilyl chloride (1.92 g, 12.80 mmol) was added to a solution of compound 10-1 (0.50 mL, 6.40 mmol) and imidazole (1.31 g, 19.2 mmol) in DMF (10 mL). The reaction mixture was stirred at 20°C for 16 hours. After completion of the reaction, the reaction mixture was quenched with water (100 mL) and extracted with ethyl acetate (50 mL x 2). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product, which was then isolated and purified by column chromatography (eluted with petroleum ether) to obtain compound 10-2.

¹ H NMR (400MHz, CDCl ₃ ): δ3.74 (t, J=6.4Hz, 2H), 2.68-2.59 (m, 2H), 1.57 (s, 1H), 0.93-0.90 (m, 9H), 0.10-0.07 (m, 6H).

Preparation of compound 10-3

Under nitrogen atmosphere, tris(dibenzylideneacetone)dipalladium (31.0 mg, 0.03 mmol) and 4,5-bis(diphenylphosphino-9,9-dimethylxanthene) (39.1 mg, 0.07 mmol) were added to a solution of compound 3-1 (200 mg, 0.34 mmol), compound 10-2 (130 mg, 0.68 mmol), and N,N-diisopropylethylamine (0.3 mL, 2.03 mmol) in 1,4-dioxane (4 mL). The reaction solution was stirred at 100°C for 16 hours. After completion of the reaction, the mixture was cooled to room temperature, quenched with water (50 mL), and extracted with ethyl acetate (20 mL × 2). The combined organic phases were washed with brine (20 mL), separated, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product, which was separated and purified by column chromatography (petroleum ether/ethyl acetate = 10/1 to 2/1) to obtain compound 10-3.

¹ H NMR (400MHz, CDCl ₃ ): δ＝8.71(s,1H),8.46(d,J＝5.6Hz,1H),8.16(dd,J＝2.0,5.6Hz,1H),7.95(d,J＝2.0Hz ,1H),7.85(d,J＝4.0Hz,1H),7.24-7.16(m,2H),5.66(d,J＝3.6Hz,1H),5.06(d,J＝11.2H z,1H),4.54(dd,J＝8.0,11.2Hz,1H),3.73-3.65(m,2H),3.15(dq,J＝4.8,7.2Hz,3H),3 .04-2.95(m,2H),2.83(m,1H),0.81(s,9H),0.78-0.73(m,3H),0.04(d,J=10.0Hz,6H);

m/z (ESI): [M+H] ⁺ = 634.1.

Preparation of Example 10

Under a nitrogen atmosphere, compound 10-3 (95 mg, 0.15 mmol) was added to a solution of methanolic hydrochloric acid (2N, 5 mL), and the reaction mixture was reacted at 17°C for 0.5 hour. After completion of the reaction, the reaction mixture was concentrated under reduced pressure, and the residue was diluted with methanol (2 mL), adjusted to pH 8 with aqueous ammonia, and then purified by reverse phase preparative purification (column: Waters Xbridge 150*25mm*5um, water ( _{NH4HCO3 )} -ACN, 38%-68%, 15 min) to obtain Example 10.

¹ H NMR (400MHz, DMSO-d ₆ ): δ10.68 (s, 1H), 8.48 (d, J = 5.6Hz, 1H), 8.26 (d, J = 2.0Hz, 1H), 8.09-8.03 (m, 1H), 7.83 (dd, J = 2.4, 5.6Hz, 1H), 7.62 (br d,J＝2.0Hz,1H),7.55-7.43(m,1H),7.29(dd,J＝5.2,8.0Hz,1H),5.17(d,J＝10.4Hz,1H),4.84(t,J＝5.6Hz,1H), 4.69(dd,J＝7.6,10.4Hz,1H),3.51-3.39(m,2H),2.96-2.89(m,2H),2.84(t,J＝7.2Hz,1H),1.64(s,3H),0.70(br d,J=6.0Hz,3H);

m/z(ESI):[M+H] ⁺ =520.2.

Example 11

Preparation of compound 11-2

Under nitrogen atmosphere, triethylamine (223.8 μL, 1.61 mmol) and 11-1 (81.67 mg, 0.54 mmol) were added sequentially to compound 1-15 (200 mg, 0.54 mmol) in 4 mL of dichloromethane. The reaction solution was stirred at 25°C for 1 hour. After completion of the reaction, the crude product was concentrated and purified by reverse preparative (0.1% FA) to obtain compound 11-2.

m/z(ESI):[M+H] ⁺ =489.1.

Preparation of compound 11-3

Boron tribromide (0.1 mL, 0.24 mmol) was slowly added dropwise to 2 mL of dichloromethane containing compound 11-2 (80 mg, 0.16 mmol) under a nitrogen atmosphere at zero degrees Celsius. The reaction solution was stirred at 25°C for 1.5 hours. After the reaction was completed, 3 mL of dichloromethane was added for dilution, and 4 mL of saturated sodium bicarbonate aqueous solution was added dropwise for quenching. The mixture was extracted with dichloromethane (5 mL × 3). The combined organic phases were washed with 20 mL of saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product, which was purified and isolated by reverse preparative method (0.1% FA) to obtain 11-3.

LC-MS (ESI): [M+H] ⁺ = 475.1.

Preparation of Example 11

Under a nitrogen atmosphere, 3-bromopropyne (12.54 mg, 0.11 mmol) was added to a 1 mL N,N-dimethylformamide solution containing compound 11-3 (50 mg, 0.11 mmol) and K ₂ CO ₃ (72.82 mg, 0.53 mmol). The reaction mixture was incubated at 60°C for 2 hours. After completion of the reaction, the mixture was cooled to room temperature and filtered. The filtrate was purified by reverse phase preparative purification (Waters Xbridge 150×25mm×5um, water (NH ₄ HCO ₃ )-ACN, 45%-75% over 10 min) to obtain Example 11.

¹ H NMR (400 MHz, DMSO-d ₆ ): δ10.54(s,1H),8.34(d,J＝5.52Hz,1H),7.86(d,J＝1.88Hz,1H),7.49(dd, J＝5.52,2.12Hz,1H),7.11-7.28(m,2H),5.18(s,1H),5.09(d,J＝10.52Hz,1H ),4.86-4.99(m,2H),4.33(dd,J＝10.36,7.52Hz,1H),3.65(t,J＝2.36Hz,1H ),2.84(t,J＝7.44Hz,1H),1.60(s,3H),1.39(s,6H),0.73(d,J＝6.36Hz,3H);

m/z(ESI):[M+H] ⁺ =513.1.

Example 12

Preparation of compound 12-2

Under nitrogen atmosphere, deuterated lithium aluminum hydride (2.6 mL, 5.12 mmol, 2 M THF solution) was slowly added dropwise to a solution of compound 12-1 (1 g, 6.40 mmol) in 40 mL of tetrahydrofuran at 0°C. The reaction was stirred at 0°C for 1 hour. After completion of the reaction, the reaction was quenched with 1 mL of deuterated water, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product, which was purified by column chromatography (petroleum ether:ethyl acetate = 10:1) to obtain compound 12-2.

¹ H NMR (400MHz, CDCl ₃ ): δ1.67 (s, 1H), 0.20-0.17 (m, 9H).

Preparation of compound 12-3

Under a nitrogen atmosphere, tributyl cyanomethylene phosphate (176.54 mg, 0.77 mmol) and compound 12-2 (95.28 mg, 0.73 mmol) were added sequentially to a solution of compound 1-18 (80 mg, 0.15 mmol) in 2 mL of dioxane. The reaction was stirred at 40°C for 16 hours. After completion, the reaction was quenched with 10 mL of water and extracted with ethyl acetate (10 mL x 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product, which was then purified by silica gel column chromatography (petroleum ether:ethyl acetate = 2:1) to isolate compound 12-3.

¹ H NMR (400MHz, CDCl ₃ ): δ8.71-8.67(m,1H),8.47(d,J＝5.6Hz,1H),8.18(dd,J＝2.0,5.6Hz,1H),7.92(d,J＝2.0Hz,1H),7.88(br d,J＝2.0Hz,1H),7.17-7.11(m,1H),6.99-6.93(m,1H),5.62-5.56(m,1H),5.04(d,J＝11.2Hz,1H), 4.18(dd,J＝6.8,10.8Hz,1H),2.91(t,J＝7.6Hz,1H),1.72(s,3H),0.84-0.80(m,3H),0.09(s,9H);

m/z (ESI): [M+H] ⁺ = 572.2.

Preparation of Example 12

Under a nitrogen atmosphere, tetrabutylammonium fluoride (235 μL, 0.23 mmol, 1 M THF solution) was added to a solution of compound 12-3 (60 mg, 0.08 mmol) in 2 mL of tetrahydrofuran, and the reaction mixture was stirred at 22.5°C for 1.5 hours. After completion of the reaction, the mixture was quenched with 40 mL of water and extracted with ethyl acetate (20 mL × 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product, which was then purified by reverse phase preparative purification (Column: Waters Xbridge 150 × 25 mm × 5 μm; Condition: water (NH ₄ HCO ₃ )-ACN; Begin B: 43, End B: 73; Gradient Time (min): 10; 100% B Hold Time (min): 4, Flow Rate (ml/min): 25) to obtain Example 12.

¹ H NMR (400MHz, DMSO-d ₆ ): δ10.71 (br s,1H),8.48(d,J＝5.6Hz,1H),8.26(d,J＝2.0Hz,1H),8.06(d,J＝2.0Hz,1H),7.83(dd,J＝2.0,5.6Hz,1H),7.61(d,J＝2.0Hz,1H),7.28-7 .15(m,2H),5.11(d,J＝10.2Hz,1H),4.33(dd,J＝7.2,10.2Hz,1H),3.63(s,1H),2.90-2.78(m,1H),1.62(s,3H),0.74(d,J＝6.8Hz,3H);

m/z(ESI):[M+H] ⁺ =500.1.

Example 13

Under a nitrogen atmosphere, 4-bromobutyne (193.81 mg, 1.46 mmol) and potassium carbonate (225.64 mg, 1.63 mmol) were sequentially added to a 3 mL DMF solution of compound 1-18 (150 mg, 0.33 mmol). The reaction mixture was reacted at 80°C for 16 hours. After completion of the reaction, the reaction mixture was cooled to room temperature and filtered. The filtrate was purified by reverse phase preparative purification (Waters Xbridge 150×25mm×5um, water (water ( _{NH4HCO3 )} -ACN, 48%-78%, 25 min)) to obtain Example 13.

¹ H NMR (400MHz, DMSO-d ₆ ): δ10.69(s,1H),8.49(d,J＝5.52Hz,1H),8.28(d,J＝2.00Hz,1H),8.06(s,1H),7.84( dd,J＝5.56,2.20Hz,1H),7.62(d,J＝2.24Hz,1H),7.23-7.13(m,2H),5.12(d,J＝10.52H z,1H),4.35(dd,J＝10.28,7.64Hz,1H),4.30-4.23(m,1H),4.20-4.13(m,1H),2.89-2. 87(m,1H),2.69-2.65(m,2H),2.47-2.43(m,1H),1.63(s,3H),0.72(d,J=6.76Hz,3H);

m/z(ESI):[M+H]+＝512.1.

Example 14

Preparation of compound 14-2

Under nitrogen at 0°C, sodium hydride (14.5 mg, 0.36 mmol) was slowly added to a 5 mL DMF solution of compound 14-1 (0.4 mL, 8.06 mmol). The reaction solution was stirred at 0°C for half an hour, and then propargyl bromide (642 mg, 5.40 mmol) was added dropwise. After the addition was complete, the reaction solution was warmed to 50°C and stirred for 16 hours. After the reaction was completed, the reaction solution was added dropwise to ice-cold saturated ammonium chloride (0.0 mL) to quench the reaction. The mixture was extracted with ethyl acetate (20 mL × 2). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product, which was separated and purified by silica gel column chromatography (petroleum ether/ethyl acetate = 2/1) to obtain compound 14-2.

¹ H NMR (400MHz, DMSO-d ₆ ): δ4.65 (t, J = 5.4Hz, 1H), 4.13 (d, J = 2.4Hz, 2H), 3.53-3.47 (m, 2H), 3.47-3.43 (m, 2H), 3.41 (t, J = 2.4Hz, 1H).

Preparation of Example 14

Under a nitrogen atmosphere, tributyl cyanomethylene phosphate (131 mg, 0.54 mmol) was added to a 2 mL dioxane solution containing compound 1-18 (50.0 mg, 0.11 mmol) and compound 14-2 (54.5 mg, 0.54 mmol). The reaction mixture was heated to 40°C for 2 hours. After completion, the reaction mixture was quenched with 50 mL of water and extracted with ethyl acetate (20 mL x 2). The combined organic phases were washed with brine (20 mL), and the aqueous phase was extracted again with ethyl acetate (20 mL). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by reverse phase preparative purification (column: Waters Xbridge 150×25 mm×5 μm, water (NH ₄ HCO ₃ )-ACN, 40%-70%, 15 min) to obtain Example 14.

¹ H NMR (400MHz, DMSO-d ₆ ): δ10.70(s,1H),8.49(d,J＝5.6Hz,1H),8.27(d,J＝2.0Hz,1H),8.07(d,J＝2.0Hz,1H), 7.84(dd,J＝2.0,5.6Hz,1H),7.62(d,J＝2.0Hz,1H),7.22-7.12(m,2H),5.11(d,J＝10.8H z,1H),4.36-4.27(m,2H),4.27-4.21(m,1H),4.19(t,J＝2.4Hz,2H),3.75(td,J＝2.8,5. 2Hz,2H),3.44(t,J＝2.4Hz,1H),2.91-2.80(m,1H),1.62(s,3H),0.71(d,J＝6.0Hz,3H);

m/z (ESI): [M+H] ⁺ = 542.2.

Example 15

Preparation of compound 15-2

Under a nitrogen atmosphere, n-butyllithium (6.8 mL, 17.12 mmol, 2.5 M) was slowly added dropwise to a solution of compound 15-1 (1 g, 14.27 mmol) in tetrahydrofuran (10 mL) at -78°C. The reaction solution was stirred at -78°C for 1 hour, then paraformaldehyde (2.06 g, 22.83 mmol) was added. The reaction solution was slowly warmed to 25°C and stirred for 16 hours. After completion of the reaction, the reaction solution was cooled to 0°C and quenched with saturated aqueous ammonium chloride (50 mL). The mixture was extracted with ethyl acetate (50 mL × 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product, which was then purified by silica gel column chromatography (petroleum ether/ethyl acetate = 2/1) to isolate compound 15-2.

¹ H NMR (400MHz, DMSO-d ₆ ): δ5.19 (t, J=5.2Hz, 1H), 4.12-4.08 (m, 4H), 3.25 (s, 3H).

Preparation of Example 15

Under nitrogen atmosphere, tributyl cyanomethylene phosphate (116.90 mg, 0.48 mmol) and compound 15-2 (48.49 mg, 0.48 mmol) were added sequentially to a solution of compound 1-18 (50 mg, 0.10 mmol) in dioxane (2 mL). The reaction mixture was stirred at 40°C for 12 hours. After completion of the reaction, the mixture was quenched with water (40 mL) and extracted with ethyl acetate (20 mL × 2). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was purified by reverse phase preparative chromatography (Column: Waters Xbridge 150 × 25 mm × 5 μm; water (NH ₄ HCO ₃ )-ACN; Begin B: 40, End B: 70; Gradient Time (min): 10; 100% B Hold Time (min): 4, Flow Rate (ml/min): 25) to obtain Example 15.

¹ H NMR (400 MHz, DMSO-d ₆ ): δ10.72(s,1H),8.49(d,J＝5.6Hz,1H),8.28(d,J＝2.0Hz,1H),8.07(d,J＝2.4H z,1H),7.84(dd,J＝2.0,5.6Hz,1H),7.62(d,J＝2.4Hz,1H),7.28-7.15(m,2H),5 .12(d,J＝10.4Hz,1H),5.06-4.93(m,2H),4.34(dd,J＝7.2,10.4Hz,1H),4.12-4 .01(m,2H),3.13(s,3H),2.88-2.80(m,1H),1.61(s,3H),0.73(d,J＝6.0Hz,3H);

m/z (ESI): [M+H] ⁺ = 542.2.

Example 16

Preparation of compound 16-2

Under a nitrogen atmosphere at 0°C, tert-butyldiphenylsilyl chloride (21.57 g, 78.47 mmol) was added to a solution of compound 16-1 (5 g, 71.34 mmol), 4-dimethylaminopyridine (0.87 g, 7.13 mmol), and triethylamine (19.8 mL, 142.67 mmol) in dichloromethane (50 mL). The reaction mixture was warmed to 25°C and stirred for 12 hours. After completion, the reaction was quenched with saturated aqueous ammonium chloride (50 mL). The aqueous phase was extracted with dichloromethane (50 mL × 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product, which was then isolated and purified by silica gel column chromatography (petroleum ether/ethyl acetate = 10/1) to obtain compound 16-2.

¹ H NMR (400MHz, CDCl ₃ ): δ7.77(td,J＝1.3,7.9Hz,2H),7.70(td,J＝1.3,7.9Hz,2H),7.51-7.37(m,6H),4.53-4.4 2(m,1H),2.35(dd,J＝0.6,2.0Hz,1H),1.41(dd,J＝0.9,6.4Hz,3H),1.10(d,J＝1.0Hz,9H).

Preparation of compound 16-3

Under a nitrogen atmosphere, methyl lithium etherate (15.2 mL, 24.31 mmol, 1.6 M) was slowly added dropwise to a solution of compound 16-2 (5 g, 16.21 mmol) in tetrahydrofuran (100 mL) at -70°C. The reaction solution was stirred at -70°C for 1 hour, and then paraformaldehyde (0.73 g, 24.31 mmol) was added portionwise to the reaction solution. The mixture was warmed to 22°C and stirred for 12 hours. After completion of the reaction, the reaction solution was cooled to 0°C and quenched by the dropwise addition of saturated aqueous ammonium chloride (50 mL). The aqueous phase was extracted with ethyl acetate (50 mL × 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product, which was then isolated and purified by silica gel column chromatography (petroleum ether/ethyl acetate = 10/1) to obtain compound 16-3.

¹ H NMR (400MHz, CDCl ₃ ): δ7.76(d,J＝6.6Hz,2H),7.70(d,J＝6.5Hz,2H),7.48-7.35(m,6H),4.54(q,J＝6.4 Hz,1H),4.10-4.06(m,2H),1.42(d,J=6.5Hz,3H),1.21-1.14(m,1H),1.08(s,9H).

Preparation of compound 16-4

Under nitrogen, tributyl cyanomethylene phosphate (236.44 mg, 0.98 mmol) was added to a solution of 1-18 (150 mg, 0.33 mmol) and compound 16-3 (1105.38 mg, 3.27 mmol) in 1,4-dioxane (1.5 mL). The reaction was stirred at 25°C for 12 hours. After completion of the reaction, the reaction solution was filtered, and the filtrate was purified by reverse phase column (0.1% formic acid system) to obtain compound 16-4.

¹ H NMR (400MHz, CDCl ₃ ): δ8.69(s,1H),8.42(d,J＝5.6Hz,1H),8.14(dd,J＝2.1,5.5Hz,1H),7.92(d,J＝2.0Hz,1H),7.89-7.8 4(m,1H),7.68(dd,J＝1.5,7.9Hz,2H),7.61(dd,J＝1.3,7.9Hz,2H),7.45-7.32(m,7H),7.15-7.08(m, 1H),7.00-6.90(m,1H),5.65(d,J＝4.1Hz,1H),5.01(d,J＝11.3Hz,1H),4.76(q,J＝15.1Hz,2H),4.42( q,J＝6.5Hz,1H),2.89-2.78(m,1H),1.64(s,3H),1.29-1.27(m,3H),1.01(s,9H),0.82-0.77(m,3H);

m/z (ESI): [M+H] ⁺ = 780.2.

Preparation of Example 16

Under nitrogen atmosphere, ammonium fluoride (189.98 mg, 5.13 mmol) was added to a solution of compound 16-4 (200 mg, 0.26 mmol) in methanol (4 mL), and the reaction was stirred at 25°C for 12 hours. After completion of the reaction, the reaction solution was filtered, and the filtrate was purified by reverse phase preparative purification (Phenomenex luna C18 150×40 mm×15 μm; mobile phase: water (FA)-ACN; B%: 42%-72%, 15 min) to obtain Example 16.

¹ H NMR (400 MHz, DMSO-d ₆ ): δ10.72(s,1H),8.49(d,J＝5.4Hz,1H),8.27(d,J＝1.9Hz,1H),8.07(d,J＝2.1 Hz,1H),7.84(dd,J＝2.2,5.6Hz,1H),7.62(d,J＝2.3Hz,1H),7.27-7.14(m,2H), 5.32(d,J＝5.4Hz,1H),5.12(d,J＝10.5Hz,1H),4.93(s,2H),4.40-4.22(m,2H) ,2.90-2.81(m,1H),1.62(s,3H),1.20(d,J＝6.5Hz,3H),0.74(d,J＝6.6Hz,3H);

m/z (ESI): [M+H] ⁺ = 542.2.

Example 17

Preparation of compound 17-2

Under a nitrogen atmosphere at zero degrees Celsius, tert-butyldiphenylsilyl chloride (8.63 g, 31.39 mmol) was added to a solution of compound 17-1 (2 g, 28.53 mmol), 4-dimethylaminopyridine (0.35 g, 2.85 mmol), and triethylamine (7.9 mL, 57.07 mmol) in dichloromethane (20 mL). The reaction was stirred at 25°C for 12 hours. After completion, the reaction was quenched with saturated aqueous ammonium chloride (20 mL). The aqueous phase was extracted with dichloromethane (15 mL x 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to dryness under reduced pressure to obtain the crude product, which was then isolated and purified by silica gel column chromatography (petroleum ether/ethyl acetate = 10/1) to obtain compound 17-2.

¹ H NMR (400MHz, CDCl ₃ ): δ7.77(dd,J＝1.3,7.8Hz,2H),7.73-7.68(m,2H),7.48-7.36(m,6H),4.47(dq ,J＝2.0,6.5Hz,1H),2.35(d,J＝2.1Hz,1H),1.41(d,J＝6.5Hz,3H),1.10(s,9H).

Preparation of compound 17-3

Under a nitrogen atmosphere, methyl lithium etherate (4.9 mL, 7.78 mmol, 1.6 M) was slowly added dropwise to a solution of compound 17-2 (1.6 g, 5.19 mmol) in tetrahydrofuran (30 mL) at -70°C. The reaction mixture was stirred at -70°C for 1 hour, and then paraformaldehyde (0.47 g, 15.56 mmol) was added portionwise to the reaction mixture. The mixture was warmed to 27°C and stirred for 12 hours. After completion of the reaction, the reaction mixture was cooled to 0°C and quenched by the dropwise addition of saturated aqueous ammonium chloride (15 mL). The aqueous phase was extracted with ethyl acetate (20 mL × 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to dryness under reduced pressure to obtain the crude product, which was then purified by silica gel column chromatography (petroleum ether/ethyl acetate = 10/1) to obtain compound 17-3.

¹ H NMR (400MHz, CDCl ₃ ): δ7.77(dd,J＝1.1,7.6Hz,2H),7.73-7.68(m,2H),7.49-7.35(m,6H),4.6 0-4.50(m,1H),4.09(d,J＝1.4Hz,2H),1.42(d,J＝6.5Hz,3H),1.08(s,9H).

Preparation of compound 17-4

Under nitrogen atmosphere, tributyl cyanomethylene phosphate (157.62 mg, 0.65 mmol) was added to a solution of 1-18 (100 mg, 0.22 mmol) and compound 17-3 (736.92 mg, 2.18 mmol) in 1,4-dioxane (1 mL). The reaction mixture was stirred at 25°C for 12 hours. The reaction mixture was filtered, and the filtrate was purified by reverse-phase column chromatography (0.1% formic acid) to obtain compound 17-4.

¹ H NMR (400MHz, CDCl ₃ ): δ8.61(s,1H),8.36(d,J＝5.6Hz,1H),8.08(dd,J＝2.2,5.6Hz,1H),7.85(d,J＝2.0Hz,1H),7.84-7.79(m,1H),7 .61(dd,J＝1.3,7.9Hz,2H),7.58-7.52(m,2H),7.39-7.23(m,7H),7.09-7.02(m,1H),6.94-6.83(m,1H),5.56(br d,J＝3.5Hz,1H),4.94(d,J＝11.3Hz,1H),4.80-4.58(m,2H),4.35(q,J＝6.5Hz,1H),2. 79-2.67(m,1H),1.55(s,3H),1.20(s,3H),0.95(s,9H),0.70(dd,J＝1.9,5.3Hz,3H);

m/z (ESI): [M+H] ⁺ = 780.3.

Preparation of Example 17

Under nitrogen atmosphere, ammonium fluoride (132.98 mg, 3.59 mmol) was added to a solution of compound 17-4 (140 mg, 0.18 mmol) in methanol (3 mL), and the reaction solution was stirred at 25°C for 12 hours. After completion of the reaction, the reaction solution was filtered, and the filtrate was purified by reverse phase preparative purification (Phenomenex luna C18 150*25mm*10um; mobile phase: water (FA)-ACN; B%: 42%-72%, 15 min) to obtain Example 17.

¹ H NMR (400 MHz, DMSO-d ₆ ): δ10.73(s,1H),8.49(d,J＝5.5Hz,1H),8.27(d,J＝2.0Hz,1H),8.06(d,J＝2.1 Hz,1H),7.84(dd,J＝2.1,5.5Hz,1H),7.62(d,J＝2.0Hz,1H),7.28-7.15(m,2H), 5.32(d,J＝5.4Hz,1H),5.12(d,J＝10.5Hz,1H),4.94(s,2H),4.40-4.23(m,2H) ,2.90-2.79(m,1H),1.63(s,3H),1.15(d,J＝6.6Hz,3H),0.74(d,J＝6.1Hz,3H);

m/z(ESI):[M+H] ⁺ =542.1.

Example 18

Preparation of compound 18-2

Under a nitrogen atmosphere, lithium aluminum hydride (0.34 g, 8.15 mmol) was slowly added portionwise to a solution of compound 18-1 (1 g, 10.19 mmol) in diethyl ether (15 mL) at 0°C. The reaction was stirred at 0°C for 1 hour. Deuterated water (1 mL) was added to the reaction solution for quenching, and the mixture was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product, which was then purified by silica gel column chromatography (petroleum ether/ethyl acetate = 10/1) to obtain compound 18-2.

¹ H NMR (400MHz, CDCl ₃ ): δ1.85 (s, 3H).

Preparation of Example 18

Under nitrogen atmosphere, tributyl cyanomethylene phosphate (231.18 mg, 0.96 mmol) and compound 18-2 (138.12 mg, 1.92 mmol) were added to a solution of compound 1-18 (100 mg, 0.19 mmol) in dioxane (1 mL). The reaction solution was heated to 40°C and stirred for 16 hours. After completion, water (30 mL) was added to quench the reaction and the mixture was extracted with ethyl acetate (30 mL × 2). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was then purified by reverse phase preparative purification (Column: Waters Xbridge 150×25 mm×5 μm; Condition: water (NH ₄ HCO ₃ )-ACN; Begin B: 43, End B: 73; Gradient Time (min): 15; 100% B Hold Time (min): 4, Flow Rate (ml/min): 25) to obtain Example 18.

¹ H NMR (400MHz, DMSO-d ₆ ): δ10.72(s,1H),8.49(d,J＝5.6Hz,1H),8.28(d,J＝2.0Hz,1H),8.06(d,J＝2.0Hz,1H),7.84(dd,J＝2.0,5.6Hz,1H),7.62(d,J＝2.0Hz,1H),7 .28-7.14(m,2H),5.12(d,J＝10.8Hz,1H),4.37(dd,J＝7.2,10.8Hz,1H),2.90-2.79(m,1H),1.75(s,3H),1.62(s,3H),0.73(d,J＝6.0Hz,3H);

m/z(ESI):[M+H] ⁺ =514.1.

Example 19

Preparation of compound 19-2

Under a nitrogen atmosphere, n-butyllithium (14.1 mL, 35.2 mmol, 2.5 M) was slowly added dropwise to a solution of compound 19-1 (5.00 g, 29.4 mmol) in tetrahydrofuran (150 mL) at -40°C. The reaction solution was stirred at -40°C for 0.5 hours, and then chloroformate (3.3 mL, 43.2 mmol) was added dropwise. The mixture was stirred at -40°C for 2 hours. The reaction solution was slowly added dropwise to a saturated aqueous ammonium chloride solution (50 mL) precooled to 0°C. The aqueous phase was extracted with ethyl acetate (150 mL x 2). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product, which was then separated and purified by silica gel column chromatography (petroleum ether/ethyl acetate = 50/1) to obtain compound 19-2.

¹ H NMR (400MHz, CDCl ₃ ): δ4.44(s,2H), 3.79(s,3H), 0.92(s,9H), 0.14(s,6H).

Preparation of compound 19-3

Under nitrogen atmosphere, lithium aluminum deuteride (0.29 g, 7.01 mmol) was slowly added portionwise to a solution of compound 19-2 (2.00 g, 8.76 mmol) in tetrahydrofuran (100 mL) at -70 ° C. The mixture was stirred at -70 ° C for 2 hours, warmed to 0 ° C, and quenched by slowly adding 1 mL of deuterated water dropwise. After stirring for 10 minutes, the mixture was warmed to 25 ° C. Anhydrous sodium sulfate was added to dryness, filtered, and the filtrate was concentrated under reduced pressure to dryness to obtain a crude product, which was purified by silica gel column chromatography (petroleum ether/ethyl acetate = 10/1) to obtain compound 19-3.

¹ H NMR (400MHz, DMSO-d ₆ ): δ5.12 (s, 1H), 4.32 (s, 2H), 0.87 (s, 9H), 0.08 (s, 6H).

Preparation of compound 19-4

Under a nitrogen atmosphere, methanesulfonic anhydride (194 mg, 1.11 mmol) was slowly added to a solution of compound 19-3 (300 mg, 1.48 mmol) and triethylamine (0.6 mL, 4.45 mmol) in dichloromethane (3 mL). The reaction was stirred at 25°C for 2 hours. After completion, 20 mL of water was added to quench the reaction. The aqueous phase was extracted with dichloromethane (10 mL x 2). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain compound 19-4.

¹ H NMR (400MHz, DMSO-d ₆ ) δ = 4.40 (s, 2H), 3.23 (s, 3H), 0.87 (s, 10H), 0.09 (s, 6H).

Preparation of compound 19-5

Under nitrogen atmosphere, compound 19-4 (183 mg, 0.65 mmol) was added to a solution of compound 1-18 (100 mg, 0.22 mmol) and potassium carbonate (150 mg, 1.09 mmol) in N-N, dimethylformamide (2 mL), and the reaction solution was stirred at 60 ° C for 12 hours. After the reaction was completed, the reaction solution was cooled to room temperature and diluted with 20 mL of water. The aqueous phase was extracted with ethyl acetate (10 mL × 2). The combined organic phases were dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated to dryness under reduced pressure to obtain the crude product, which was separated and purified by reverse phase preparative (0.1% FA/ACN, 80% to 90% over 10 min) to obtain compound 19-5.

¹ H NMR (400MHz, CDCl ₃ ): δ8.66(s,1H),8.46(d,J＝5.6Hz,1H),8.16(dd,J＝2.4,5.6Hz,1H),7.92(d,J ＝2.0Hz,1H),7.88-7.81(m,1H),7.16-7.10(m,1H),7.03-6.91(m,1H),5.58(br s,1H),5.02(d,J＝11.2Hz,1H),4.21(d,J＝1.6Hz,2H),2.93-2.84(m,1H),1.71(s,3H),0.85(s,9H),0.83-0.78(m,3H),0.02(s,6H);

m/z (ESI): [M+H] ⁺ = 644.3.

Preparation of Example 19

Under nitrogen atmosphere, 3 mL of hydrochloric acid/methanol (2 M) solution containing compound 19-5 (60.0 mg, 0.09 mmol) was stirred at 25°C for 0.5 hours. After the reaction was completed, the solution was concentrated under reduced pressure to obtain the crude product, which was separated and purified by reverse phase preparation (column: Waters Xbridge 150×25mm×5um, 30% to 60% over 10 min) to obtain Example 19.

¹ H NMR (400 MHz, DMSO-d ₆ ): δ10.72(s,1H),8.49(d,J＝5.6Hz,1H),8.26(d,J＝2.0Hz,1H),8.07(d,J＝2.4 Hz,1H),7.84(dd,J＝2.4,5.6Hz,1H),7.62(d,J＝2.0Hz,1H),7.26-7.14(m,2H) ,5.19(t,J＝6.0Hz,1H),5.11(d,J＝10.4Hz,1H),4.34(dd,J＝7.6,10.4Hz,1H), 4.06(d,J＝4.0Hz,2H),2.92-2.80(m,1H),1.62(s,3H),0.74(d,J＝6.0Hz,3H);

m/z(ESI):[M+H] ⁺ =530.1.

Example 20

Preparation of Example 20

Under nitrogen atmosphere, deuterated water (0.2 mL, 9.30 mmol) and potassium carbonate (15.42 mg, 0.11 mmol) were added to 1 mL of acetonitrile solution containing 50 mg of Example 12 (0.09 mmol), and the reaction solution was stirred at 24°C for 12 hours. After the reaction was completed, the reaction solution was filtered, and the filtrate was separated and purified by reverse phase preparation (column: Phenomenex 150×25 mm×10 μm, Condition: water (FA)-CAN; Begin B: 48, End B: 78, Gradient Time (min): 11, 100% B Hold Time (min): 4, Flow Rate (ml/min): 2) to obtain Example 20.

¹ H NMR (400MHz, DMSO-d ₆ ): δ10.71(s,1H),8.49(d,J＝5.4Hz,1H),8.27(d,J＝2.0Hz,1H),8.06(d,J＝2.4Hz,1H),7.83(dd,J＝2.4,5.4Hz,1H),7.62(d,J＝2.0Hz, 1H),7.28-7.14(m,2H),5.12(d,J＝10.2Hz,1H),4.33(dd,J＝7.6,10.2Hz,1H),2.90-2.78(m,1H),1.62(s,3H),0.74(d,J＝6.0Hz,3H);

m/z(ESI):[M+H] ⁺ =501.1.

Example 21

Preparation of compound 21-1

Under nitrogen atmosphere, compound 1-15 (200 mg, 0.54 mmol) was added to methanol (2 mL), and the mixture was stirred at 25°C for 2 hours. The reaction solution was concentrated to dryness to obtain compound 21-1.

m/z (ESI): [M+H] ⁺ = 369.0.

Preparation of compound 21-2

Under a nitrogen atmosphere, boron tribromide (2.04 g, 8.15 mmol) was added dropwise to a solution of compound 21-1 (2.00 g, 5.43 mmol) in dichloromethane (40 mL) at 0°C, and the mixture was stirred at 25°C for 1.5 hours. Saturated aqueous sodium bicarbonate (20 mL) was slowly added dropwise to the reaction solution, and the mixture was extracted with dichloromethane (30 mL x 3). The combined organic phases were washed with saturated brine (20 mL x 2), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product, which was then isolated and purified by silica gel column chromatography (petroleum ether/ethyl acetate = 5/1) to obtain compound 21-2.

m/z(ESI):[M+H] ⁺ =355.1.

Preparation of compound 21-3

Propargyl bromide (0.34 g, 2.82 mmol) was added to a suspension of compound 21-2 (1.00 g, 2.82 mmol) and potassium carbonate (1.95 g, 14.1 L) in DMF (10 mL). The mixture was stirred at 60°C for 2 hours. The reaction mixture was filtered, and the filtrate was purified by reverse-phase column chromatography (0.1% formic acid) to obtain compound 21-3.

m/z(ESI):[M+H] ⁺ =393.1.

Preparation of compound 21-4

To a solution of compound 21-3 (930 mg, 2.37 mmol) in THF (12 mL), methanol (4 mL), and water (4 mL) was added lithium hydroxide (497.33 mg, 11.85 mmol). The mixture was stirred at 25°C for 1 hour. The reaction mixture was diluted with water (30 mL) and the pH was adjusted to 4 with dilute hydrochloric acid (1 mol/L). The mixture was extracted with dichloromethane (30 mL x 3). The combined organic phases were washed with saturated brine (20 mL x 2), dried over anhydrous sodium sulfate, filtered, and concentrated to yield compound 21-4.

m/z(ESI):[MH] ^- =377.0.

Preparation of compound 21-5

To a solution of compound 21-4 (70 mg, 0.19 mmol) in DCM (2 mL) were added DMF (2.9 μL, 0.04 mmol) and oxalyl chloride (31.8 μL, 0.37 mmol). The mixture was stirred at 25°C for 1 hour. The mixture was concentrated to give compound 21-5.

Preparation of compound 21-6

Preparation of compound 21-8

Under a nitrogen atmosphere, di-tert-butyl dicarbonate (2.8 mL, 13.1 mmol) and triethylamine (1.8 mL, 13.1 mmol) were added to a solution of compound 21-7 (1.00 g, 6.57 mmol) and 4-dimethylaminopyridine (0.40 g, 3.29 mmol) in 10 mL of dichloromethane. The reaction was stirred at 26°C for 16 hours. Additional di-tert-butyl dicarbonate (2.8 mL, 13.1 mmol) was added, and the reaction mixture was warmed to 40°C and stirred for another 16 hours. After completion of the reaction, the mixture was diluted with 200 mL of water, separated, and the aqueous phase was extracted with ethyl acetate (100 mL x 2). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product, which was then isolated and purified by silica gel column chromatography (petroleum ether/ethyl acetate = 10/1 to 5/1) to obtain compound 21-8.

¹ H NMR (400MHz, CDCl ₃ ): δ8.57 (d, J = 5.6 Hz, 1H), 8.03 (d, J = 2.4 Hz, 1H), 7.71 (dd, J = 2.0, 5.6 Hz, 1H), 7.00 (s, 1H), 4.00 (s, 3H), 1.54 (s, 9H);

m/z (ESI): [M+H] ⁺ = 253.0.

Preparation of compound 21-9

At 0°C, m-chloroperbenzoic acid (965.69 mg, 4.76 mmol, 85% content) was slowly added portionwise to a solution of compound 21-8 (600 mg, 2.38 mmol) in 10 mL of dichloromethane. The reaction mixture was heated to 26°C and stirred for 16 hours. After completion of the reaction, the reaction mixture was filtered, and the filtrate was slowly added dropwise to 50 mL of saturated sodium sulfite aqueous solution to quench the reaction. After a negative result was obtained using starch potassium iodide test paper, the aqueous phase was extracted with dichloromethane (20 mL x 2). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. Compound 21-9 was obtained by silica gel column chromatography (petroleum ether/ethyl acetate = 10/1 to 100% methanol).

¹ H NMR (400MHz, CDCl ₃ ): δ8.13 (d, J = 7.2 Hz, 1H), 7.86 (s, 1H), 7.53 (d, J = 4.8 Hz, 1H), 3.96 (s, 3H), 1.51 (s, 9H);

m/z (ESI): [M+H] ⁺ = 269.0.

Preparation of compound 21-6

Under nitrogen atmosphere, trifluoroacetic acid (2.00 mL, 26.9 mmol) was slowly added dropwise to a 2 mL dichloromethane solution of compound 21-9 (170 mg, 0.63 mmol). The mixture was stirred at 25 °C for 2 hours and then concentrated. The pH of the residue was adjusted to 10 with triethylamine and purified by reverse phase preparation (0.1% FA/ACN, 100% water over 3 min) to obtain compound 21-6.

Preparation of compound 21-10

Under nitrogen atmosphere, compound 21-5 (70.0 mg, 0.18 mmol) was added to a 0.5 mL dichloromethane solution of compound 21-6 (148 mg, 0.88 mmol) and triethylamine (0.1 mL, 0.88 mmol), and the reaction solution was stirred at 25°C for 1.5 hours. After the reaction was completed, the reaction solution was quenched with 20 mL of water, and the aqueous phase was extracted with dichloromethane (10 mL × 2). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product, which was separated and purified by reverse phase preparative (0.1% FA/ACN, 50% to 60% over 5 min) to obtain compound 21-10.

m/z (ESI): [M+H] ⁺ = 529.2.

Preparation of Example 21

Under nitrogen atmosphere, compound 21-10 (70.0 mg, 0.13 mmol) was added to 10 mL of ammonia methanol (7 M). The reaction solution was stirred at 25 ° C for 16 hours and then concentrated to obtain the crude product. The crude product was separated and purified by reverse phase preparation (column: Waters Xbridge 150×25mm×5um, 32% to 62% over 11min) to obtain Example 21.

¹ H NMR (400 MHz, DMSO-d ₆ ): δ＝10.78(s,1H),10.59(d,J＝4.4Hz,1H),8.52(d,J＝3.2Hz,1H),8.31(d,J＝ 7.2Hz,1H),8.23(d,J＝4.4Hz,1H),7.88(dd,J＝3.2,7.2Hz,1H),7.26-7.16(m, 2H),5.10(d,J＝10.4Hz,1H),4.98-4.85(m,2H),4.32(dd,J＝7.6,10.4Hz,1H), 3.63(t,J＝2.4Hz,1H),2.88-2.66(m,1H),1.62(s,3H),0.73(d,J＝6.0Hz,3H);

m/z(ESI):[M+H] ⁺ =514.1.

Examples 22 and 23

Preparation of compound 22-2

Under nitrogen atmosphere, cesium carbonate (20.55 g, 63.08 mmol) and [1,1-bis(diphenylphosphino)ferrocene]dichloropalladium dichloromethane (1.29 g, 1.58 mmol) were added to a mixed solution of 150 mL 2-methyltetrahydrofuran and 15 mL water containing compound 22-1 (5 g, 31.54 mmol) and potassium ethylene trifluoroborate (5.07 g, 37.85 mmol). The reaction solution was heated to 90 ° C and stirred for 4 hours. After the reaction was completed, the reaction solution was cooled to room temperature and concentrated. 50 mL of water was added to the residue and diluted. The mixture was extracted with ethyl acetate (50 mL × 3). The combined organic phases were washed with 100 mL of saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by reverse phase preparative (0.1% FA system) and silica gel column chromatography (petroleum ether/ethyl acetate = 5/1 to 2/1) to obtain compound 22-2.

¹ H NMR (400MHz, CDCl ₃ ): δ9.39(d,J＝2.4Hz,1H),8.44(dd,J＝2.4,8.4Hz,1H),7.48(d,J＝8.4Hz,1H), 6.91(dd,J＝10.8,17.6Hz,1H), 6.46(d,J＝17.6Hz,1H), 5.75(d,J＝10.8Hz,1H).

Preparation of compound 22-3

Under a nitrogen atmosphere, osmium tetroxide (63.55 mg, 0.25 mmol) in 4.3 mL of aqueous solution was slowly added dropwise to a mixture of N-methylmorpholine oxide (4353.92 mg, 37.17 mmol) and compound 22-2 (1860.00 mg, 12.39 mmol) in 4.3 mL of water and 16 mL of acetone. After the addition was complete, the reaction solution was stirred at 25°C for 3 hours. After completion of the reaction, the reaction solution was cooled to zero degrees Celsius and quenched by the addition of 50 mL of saturated sodium sulfite aqueous solution. After stirring for 15 minutes, the aqueous phase was extracted with ethyl acetate (50 mL x 15). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product, which was then purified and isolated by silica gel column chromatography (petroleum ether/ethyl acetate = 5/1 to 0/1) to obtain compound 22-3.

¹ H NMR (400MHz, DMSO-d ₆ ): δ9.29(d,J＝2.4Hz,1H),8.58(dd,J＝2.8,8.8Hz,1H),7.77(d,J＝8.8Hz,1H),5.76(d,J＝5.2 Hz,1H),4.79(t,J＝6.0Hz,1H),4.72(q,J＝5.2Hz,1H),3.78-3.69(m,1H),3.62-3.53(m,1H).

Preparation of compound 22-4

Under nitrogen atmosphere, p-toluenesulfonic acid (120 mg, 0.67 mmol) and 2,2-dimethoxypropane (2.1 mL, 16.70 mmol) were added to a mixed solution of compound 22-3 (1230.00 mg, 6.68 mmol) in 20 mL of 2-methyltetrahydrofuran and 20 mL of acetone. The reaction solution was stirred at 25 ° C for 16 hours. After the reaction was completed, 10 mL of saturated sodium bicarbonate aqueous solution was added to quench the reaction, and 50 mL of water was added to dilute it. The aqueous phase was extracted with ethyl acetate (50 mL × 3), and the combined organic phases were washed with 100 mL of saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product, which was separated and purified by silica gel column chromatography (petroleum ether/ethyl acetate = 5/1) to obtain compound 22-4.

¹ H NMR (400MHz, DMSO-d ₆ ): δ9.32(d,J＝2.4Hz,1H),8.63(dd,J＝2.4,8.4Hz,1H),7.76(d,J＝8.8Hz,1H),5.27(t,J＝6.4 Hz,1H),4.45(dd,J＝7.2,8.4Hz,1H),3.91(dd,J＝6.0,8.4Hz,1H),1.46(s,3H),1.43(s,3H);

m/z (ESI): [M+H] ⁺ = 224.9.

Preparation of compound 22-5

Under an argon atmosphere, 10% palladium-on-carbon catalyst (wet) (100 mg) was added to a solution of compound 22-4 (700 mg, 3.12 mmol) in 28 mL of ethyl acetate. After hydrogen replacement three times, the reaction solution was stirred at 25°C under a hydrogen atmosphere (15 psi) for 6 hours. After completion of the reaction, the reaction solution was filtered through celite, and the filtrate was concentrated under reduced pressure to obtain compound 22-5.

Preparation of compound 22-6

Under a nitrogen atmosphere, triethylamine (559.4 μL, 4.02 mmol) was added to a 10 mL dichloromethane solution containing compound 1-15 (500 mg, 1.34 mmol) and compound 22-5 (260.56 mg, 1.34 mmol). The reaction mixture was allowed to react at 25°C for 1 hour. After completion of the reaction, 10 mL of water was added to dilute the reaction mixture. The aqueous phase was extracted with dichloromethane (15 mL × 3). The combined organic phases were washed with 30 mL of saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product, which was then separated and purified by silica gel column chromatography (petroleum ether/ethyl acetate = 5/1 to 2/1) to obtain compound 22-6.

¹ H NMR (400MHz, DMSO-d ₆ ): δ10.44(s,1H),8.70(dd,J＝2.4,5.2Hz,1H),8.05(ddd,J＝2.4,5.2,8.4Hz,1H),7 .44(d,J＝8.4Hz,1H),7.22-7.10(m,2H),5.17-4.99(m,2H),4.32(dd,J＝6.8,8.0Hz, 1H), 4.23 (dd, J＝7.6, 10.4Hz, 1H), 3.94 (d, J＝2.0Hz, 3H), 3.81 (dd, J＝6.8, 8.0Hz, 1H ),2.79-2.72(m,1H),1.60(s,3H),1.41(s,3H),1.38(s,3H),0.73(d,J＝6.0Hz,3H);

m/z (ESI): [M+H] ⁺ = 531.2.

Preparation of compound 22-7

Under a nitrogen atmosphere at zero degrees Celsius, boron tribromide (0.8 mL, 1.67 mmol) was slowly added to compound 22-6 (590 mg, 1.11 mmol) in 12 mL of dichloromethane. The reaction mixture was heated to 25°C and stirred for 1.5 hours. After completion, the reaction was quenched by the addition of 10 mL of saturated sodium bicarbonate solution. The aqueous phase was extracted with dichloromethane (20 mL × 3). The combined organic phases were washed with 40 mL of saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The product was then separated and purified by silica gel column chromatography (petroleum ether/ethyl acetate = 5/1) and reverse phase column chromatography (0.1% formic acid system) to obtain compound 22-7.

¹ H NMR (400MHz, DMSO-d6): δ10.49 (br s,1H),8.66(dd,J＝2.4,9.2Hz,1H),8.35(s,1H),7.99(dt,J＝2.4,8.8Hz,1H),7.42(d,J＝ 8.4Hz,1H),7.01(t,J＝6.8Hz,1H),6.80(q,J＝8.8Hz,1H),5.57-5.20(m,1H),5.10(d,J＝1 0.4Hz,1H),4.89-4.59(m,1H),4.53(dd,J＝4.4,6.8Hz,1H),4.22(dd,J＝7.6,10.0Hz,1H) ,3.62(dd,J＝4.0,10.8Hz,2H),2.83(t,J＝7.2Hz,1H),1.58(s,3H),0.70(d,J＝6.0Hz,3H);

m/z (ESI): [M+H] ⁺ = 477.2.

Preparation of Examples 22 and 23

Under nitrogen atmosphere, propargyl bromide (32.6 μL, 0.38 mmol) was slowly added to a 1.5 mL NN, dimethylformamide solution of compound 22-7 (90 mg, 0.19 mmol) and potassium carbonate (130.54 mg, 0.94 mmol). The reaction solution was heated to 60° C. and stirred for 2 hours. After completion of the reaction, the reaction solution was cooled to room temperature and filtered. The filtrate was purified by reverse phase preparative purification (Waters Xbridge 150×25 mm×5 um; mobile phase: water _{( NH4HCO3} )-ACN; B%: 40%-60%, 25 min) to obtain a mixture of Example 22 and Example _23. The mixture was separated by SFC (Column: DAICEL CHIRALCEL OX (250 mm×30 mm×10 um); Condition: CO2 _- EtOH (0.1% _NH3H2O ); B%: 15-15; Gradient Time (min): 6.2; 100% B Hold Time (min): 0; Flow Rate (ml/min): 150) to obtain two single isomers.

Example 22 (retention time = 1.505 minutes):

¹ H NMR (400MHz, DMSO-d ₆ ): δ10.36(s,1H),8.63(d,J＝2.4Hz,1H),7.99(dd,J＝2.4,8.4Hz,1H),7.42(d,J＝8.4Hz,1H),7.27-7.15 (m,2H),5.33(d,J＝4.4Hz,1H),5.10(d,J＝10.4Hz,1H),4.92(dd,J＝2.4,4.0Hz,2H),4.64(t,J＝5.6Hz,1H ), 4.53(td,J＝4.4,6.8Hz,1H), 4.31(dd,J＝7.6,10.4Hz,1H), 3.66(t,J＝2.4Hz,1H), 3.62(ddd,J＝4.4,6. 0,10.8Hz,1H),3.43(td,J＝6.4,11.2Hz,1H),2.83(t,J＝7.6Hz,1H),1.60(s,3H),0.74(d,J＝6.0Hz,3H);

m/z (ESI): [M+H] ⁺ = 515.3.

Example 23 (retention time = 1.649 minutes):

¹ H NMR (400MHz, DMSO-d6) δ = 10.35 (s, 1H), 8.66 (d, J = 2.4Hz, 1H), 7.97 (dd, J = 2.4, 8.4Hz, 1H), 7.42 (d, J = 8.4 Hz,1H),7.26-7.15(m,2H),5.33(d,J＝4.8Hz,1H),5.10(d,J＝10.4Hz,1H),4.97-4.86(m,2H),4.64(t,J＝5. 6Hz, 1H), 4.53 (td, J＝4.4, 6.8Hz, 1H), 4.31 (dd, J＝7.6, 10.4Hz, 1H), 3.66 (t, J＝2.4Hz, 1H), 3.62 (ddd, J＝4 .4,6.4,10.7Hz,1H),3.43(td,J＝6.4,10.8Hz,1H),2.89-2.77(m,1H),1.61(s,3H),0.74(d,J＝5.8Hz,3H);

m/z (ESI): [M+H] ⁺ = 515.2.

Examples 24 and 25

Preparation of compound 24-1

Under a nitrogen atmosphere, cyanomethylenetri-n-butylphosphine (7.15 g, 29.64 mmol) was added to a solution of compound 21-2 (2.10 g, 5.93 mmol) and compound 12-2 (3.86 g, 29.64 mmol) in 80 mL of dioxane. The reaction mixture was heated to 40°C and stirred for 16 hours. After completion of the reaction, the mixture was cooled to room temperature and quenched by the addition of 100 mL of water. The aqueous phase was extracted with ethyl acetate (50 mL x 2). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product, which was then purified by silica gel column chromatography (petroleum ether/ethyl acetate = 50/1 to 30/1) to obtain compound 24-1.

¹ H NMR (400MHz, CDCl ₃ ): δ6.98-6.92(m,1H),6.92-6.86(m,1H),4.92(d,J＝10.4Hz,1H),4.28-4.19(m,1 H),3.71(s,3H),2.91-2.69(m,1H),1.65(s,3H),0.81-0.74(m,3H),0.15(s,9H).

Preparation of compound 24-2

Under a nitrogen atmosphere, a solution of lithium hydroxide (1.01 g, 24.1 mmol) in 8 mL of water was added dropwise to a solution of compound 24-1 (2.25 g, 4.82 mmol) in 24 mL of tetrahydrofuran and 8 mL of methanol. The reaction mixture was stirred at 26°C for 1 hour. After completion of the reaction, the mixture was concentrated under reduced pressure to remove the low-boiling solvent. The pH of the residue was adjusted to 5 with 2M dilute hydrochloric acid. The aqueous phase was extracted with ethyl acetate (20 mL x 2). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product, which was then purified by silica gel column chromatography (petroleum ether/ethyl acetate = 10/1 to ethyl acetate/methanol = 5/1) to obtain compound 24-2.

¹ H NMR (400MHz, DMSO-d ₆ ): δ7.24-7.08(m,2H),4.76(d,J＝10.4Hz,1H),4.16-4.05(m,1H),3.64(s,1H),2.74-2.66(m,1H),1.51(s,3H),0.67(d,J＝6.0Hz,3H);

m/z(ESI):[MH] ^- ＝379.1.

Preparation of compound 24-3

Under nitrogen atmosphere, oxalyl chloride (67.7 μL, 0.79 mmol) was slowly added dropwise to a 2 mL dichloromethane solution of compound 24-2 (100 mg, 0.26 mmol) and N,N-dimethylformamide (2.0 μL, 0.03 mmol). The reaction mixture was reacted at 26°C for 2 hours. After completion of the reaction, the mixture was concentrated under reduced pressure to obtain compound 24-3.

Preparation of compound 24-5

Under a nitrogen atmosphere, a solution of compound 24-3 (100 mg, 0.25 mmol) in 1 mL of dichloromethane was added dropwise to a solution of compound 24-4 (70.3 mg, 0.5 mmol) and triethylamine (0.1 mL, 0.75 mmol) in 1 mL of dichloromethane. The reaction mixture was stirred at 26°C for 1 hour. After completion of the reaction, the mixture was diluted with 50 mL of water, and the aqueous phase was extracted with dichloromethane (20 mL x 2). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product, which was then purified by silica gel column chromatography (petroleum ether/ethyl acetate = 10/1 to 5/1) to obtain compound 24-5.

¹ H NMR (400MHz, CDCl ₃ ): δ8.39-8.28(m,2H),7.52(d,J＝2.0Hz,1H),7.16-7.11(m,1H),7.09(dd,J＝2.0,5.6Hz,1H),7.03-6.95(m,1H),4 .99(d,J＝11.2Hz,1H),4.20(s,1H),2.88-2.76(m,1H),2.54(s,3H),2.37(s,1H),1.70(s,3H),0.84-0.76(m,3H);

m/z (ESI): [M+H] ⁺ = 503.2.

Preparation of Examples 24 and 25

Under nitrogen atmosphere, iodobenzene diacetate (216.7 mg, 0.67 mmol) was added to a 4 mL methanol solution of compound 24-5 (160 mg, 0.32 mmol) and ammonium acetate (39.27 mg, 0.51 mmol), and the reaction solution was stirred at 26°C for 1 hour. After completion of the reaction, the product was diluted with 20 mL of water and concentrated under reduced pressure to remove the low-boiling solvent. The aqueous phase was extracted with ethyl acetate (10 mL × 2). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product, which was purified by reverse phase preparative (0.1% _NH4OH , 40% to 50% over 10 min) and then separated by SFC (column: DAICEL CHIRALPAK AD (250 mm × 30 mm, 10 μm), CO2 _- EtOH (0.1% _NH3 · _H2O ), 25% over 4.1 min, flow rate: 60 mL/min) to obtain two single isomers.

Example 24 (retention time = 1.265 minutes):

¹ H NMR (400MHz, DMSO-d ₆ ): δ10.87(s,1H),8.57(d,J＝5.6Hz,1H),8.35(d,J＝2.0Hz,1H),7.82(dd,J＝2.0,5.6Hz,1H),7.30-7.13(m,2H),5.13( d,J＝10.4Hz,1H),4.39-4.27(m,2H),3.63(s,1H),3.11(s,3H),2.89-2.78(m,1H),1.61(s,3H),0.74(d,J＝6.4Hz,3H);

m/z(ESI):[M+H] ⁺ =534.1.

Example 25 (retention time = 1.412 minutes):

¹ H NMR (400MHz, DMSO-d6): δ10.87(s,1H),8.57(d,J＝5.6Hz,1H),8.35(d,J＝2.0Hz,1H),7.82(dd,J＝2.0,5.6Hz,1H),7.29-7.14(m,2H), 5.13(d,J＝10.4Hz,1H),4.39-4.28(m,2H),3.63(s,1H),3.11(d,J＝1.2Hz,3H),2.90-2.78(m,1H),1.61(s,3H),0.74(d,J＝6.0Hz,3H);

m/z(ESI):[M+H] ⁺ =534.1.

Example 26

Preparation of compound 26-2

Under a nitrogen atmosphere, compound 26-1 (380 mg, 2.55 mmol) was added to 10 mL of a 6 M ammonia methanol solution, and the reaction mixture was stirred at 25°C for 20 hours. The reaction mixture was concentrated under reduced pressure to obtain a crude product, which was then separated and purified by silica gel column chromatography (petroleum ether/ethyl acetate = 1/1 to 1/1, dichloromethane/methanol = 20/1 to 10/1) to obtain compound 26-2.

¹ H NMR (400MHz, DMSO-d ₆ ): δ7.83(brs,1H),7.32(brs,1H),7.06(d,J＝8.0Hz,1H),6.73(d,J＝2.4Hz,1H),6.5 8(dd,J＝2.4,8.0Hz,1H),5.16(s,2H),5.07(t,J＝5.6Hz,1H),4.36(d,J＝5.6Hz,2H).

Preparation of compound 26-3

Under nitrogen atmosphere, imidazole (40.97 mg, 0.60 mmol) and tert-butyldimethylsilyl chloride (68.02 mg, 0.45 mmol) were added to a solution of compound 26-2 (50.0 mg, 0.30 mmol) in 1 mL of N,N-dimethylformamide, and the reaction mixture was reacted at 25°C for 12 hours. After completion of the reaction, 20 mL of water was added to quench the reaction, and the aqueous phase was extracted with ethyl acetate (20 mL × 2). The combined organic phases were washed with 20 mL of saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product, which was separated and purified by silica gel column chromatography (petroleum ether/ethyl acetate = 5/1 to 0/1) to obtain compound 26-3.

¹ H NMR (400MHz, DMSO-d ₆ ): δ7.61(brs,1H),7.21(brs,1H),7.09(d,J＝8.0Hz,1H),6.67(d,J＝2.4Hz,1H), 6.58(dd,J＝2.4,8.0Hz,1H),5.14(s,2H),4.65(s,2H),0.86(s,9H),0.04(s,6H).

Preparation of compound 26-4

Under a nitrogen atmosphere, compound 26-3 (53.97 mg, 0.19 mmol) and triethylamine (87.1 μL, 0.63 mmol) were added to a 2 mL dichloromethane solution of compound 24-3 (50.0 mg, 0.13 mmol). The reaction mixture was allowed to react at 25°C for 1 hour. After completion, 20 mL of water was added to quench the reaction. The aqueous phase was extracted with dichloromethane (20 mL × 2). The combined organic phases were washed with 20 mL of saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product, which was then separated and purified by silica gel column chromatography (petroleum ether/ethyl acetate = 5/1 to 1/1) to obtain compound 26-4.

¹ H NMR (400MHz, CDCl ₃ ): δ8.47(s,1H),8.03-7.93(m,2H),7.75(d,J＝2.4Hz,1H),7.29(s,1H),7.20-7.15(m,1H),7.02-6.94(m,1H),5.68(brs,1H),5.00(d,J＝11.2 Hz,1H),4.78(s,2H),4.25(dd,J＝7.6,11.2Hz,1H),2.88-2.75(m,1H), 2.38(s,1H),1.71(s,3H),0.88(s,9H),0.83-0.78(m,3H),0.10(s,6H);

m/z (ESI): [M+H] ⁺ = 643.3.

Preparation of Example 26

Under a nitrogen atmosphere, a tetrabutylammonium fluoride solution in tetrahydrofuran (133.4 μL, 0.13 mmol, 1 M) was added to a 1 mL tetrahydrofuran solution of compound 26-4 (60.0 mg, 0.09 mmol). The reaction mixture was allowed to react at 25°C for 0.5 h. After completion of the reaction, the crude product was concentrated and purified via reverse phase preparative chromatography (Phenomenex luna C18 150×25 mm×10 μm; mobile phase: water (FA)-ACN; B%: 45%-75%, 25 min) and secondary reverse phase preparative chromatography (Waters Xbridge 150×25 mm×5 μm; mobile phase: water (NH ₄ HCO ₃ )-ACN; B%: 42%-72%, 25 min) to afford Example 26.

¹ H NMR (400MHz, DMSO-d ₆ ): δ10.29(s,1H),7.86(brs,1H),7.72(d,J＝2.0Hz,1H),7.65(dd,J＝2.0,8.0Hz,1H),7.48-7.37(m,2H),7.27-7.12(m,2H),5.24(brs,1H),5.07 (d,J＝10.4Hz,1H),4.53(d,J＝3.6Hz,2H),4.32(dd,J＝7.6,10.4Hz,1H),3.64(s,1H),2.83(t,J＝7.2Hz,1H),1.60(s,3H),0.73(d,J＝6.0Hz,3H);

m/z(ESI):[M-17] ⁺ =511.1.

Example 27

Preparation of compound 27-1

Under a nitrogen atmosphere, triethylamine (365.5 μL, 2.63 mmol) was added to a solution of compound 24-3 (103.67 mg, 0.26 mmol) in dichloromethane (1 mL). A solution of methyl 4-aminopyridine-2-carboxylate (80.01 mg, 0.53 mmol) in dichloromethane (0.5 mL) was then added dropwise. The reaction mixture was stirred at 26°C for 2 hours. After completion of the reaction, the mixture was diluted with 30 mL of water, and the aqueous phase was extracted with dichloromethane (10 mL x 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product, which was then separated and purified by silica gel column chromatography (petroleum ether/ethyl acetate = 1/1) to obtain compound 27-1.

¹ H NMR (400MHz, CDCl ₃ ): δ8.63(d,J＝5.6Hz,1H),8.58(s,1H),8.08(d,J＝2.0Hz,1H),7.93(dd,J＝5.6,2.0Hz,1H),7.05-7.19(m,1H),6.93-7.05(m, 1H),5.03(d,J＝11.2Hz,1H),4.21-4.32(m,1H),4.01(s,3H),2.73-2.94(m,1H),2.38(s,1H),1.72(s,3H),0.76-0.87(m,3H);

m/z(ESI):[MH] ^- ＝513.2.

Preparation of Example 27

Under nitrogen atmosphere, potassium cyanide (2 mg, 0.03 mmol) and hydroxylamine aqueous solution (0.5 mL, 8.16 mmol, 50% aqueous solution) were added sequentially to a mixed solution of compound 27-1 (30 mg, 0.06 mmol) in 0.5 mL of methanol and 0.5 mL of tetrahydrofuran. The reaction solution was stirred at 26°C for 12 hours. After completion of the reaction, the reaction solution was poured into 20 mL of saturated sodium bicarbonate aqueous solution to quench the reaction. The mixture was extracted with dichloromethane (10 mL x 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product, which was purified by reverse phase preparation (Column Phenomenex luna C18 150*25 mm*10 um; mobile phase: water (TFA)-ACN; B%: 28%-58%, 14 min) to obtain Example 27.

¹ H NMR (400MHz, DMSO-d ₆ ): δ11.38(s,1H),10.70(s,1H),8.45(d,J＝5.6Hz,1H),8.22(d,J＝2.0Hz,1H),7.81(dd,J＝5.6,2.0Hz,1H),7.07-7. 38(m,2H),5.11(d,J＝10.4Hz,1H).4.10-4.45(m,1H),3.63(s,2H),2.84(s,1H),1.62(s,3H),0.74(d,J＝6.0Hz,3H);

m/z(ESI):[M+H] ⁺ =516.1.

Example 28

Preparation of compound 28-1

Under a nitrogen atmosphere, triethylamine (348.6 μL, 2.51 mmol) and 4-aminopyridine-2-carbonitrile (59.75 mg, 0.50 mmol) were added sequentially to a solution of compound 24-3 (100 mg, 0.25 mmol) in 1 mL of dichloromethane. The reaction mixture was stirred at 26°C for 2 hours. After completion, the reaction was diluted with 10 mL of water and extracted with dichloromethane (10 mL x 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product, which was then purified on thin-layer silica gel (petroleum ether/ethyl acetate = 1/1) to obtain compound 28-1.

¹ H NMR (400MHz, CDCl ₃ ): δ8.67-8.53(m,2H),8.08-7.99(m,1H),7.70-7.60(m,1H),7.16-7.07(m,1H),7.06-6.91(m,1H),5.13-4 .94(m,1H),4.31-4.18(m,1H),2.91-2.78(m,1H),2.44-2.36(m,1H),1.75-1.67(m,3H),0.86-0.76(m,3H);

m/z(ESI):[M+H] ⁺ =482.1.

Preparation of Example 28

Under nitrogen atmosphere, hydroxylamine hydrochloride (7.69 mg, 0.11 mmol) and sodium carbonate (23.47 mg, 0.22 mmol) were added sequentially to a 2 mL ethanol solution of compound 28-1 (41 mg, 0.09 mmol). The reaction solution was stirred at 25°C for 1 hour. After completion of the reaction, the filtrate was filtered, and the pH of the filtrate was adjusted to 5 with 2N hydrochloric acid. The product was purified by reverse phase chromatography (Column Phenomenex luna C18 150*25mm*10um Condition water (FA)-CAN Begin B 43 End B 73 Gradient Time (min) 11 100% B Hold Time (min) 4 Flow Rate (ml/min) 25) to obtain Example 28.

¹ H NMR (400MHz, DMSO-d ₆ ): δ10.74-10.50(m,1H),9.93-9.75(m,1H),8.47-8.37(m,1H),8.22-8.08(m,1H),7.69-7.56(m,1H),7.32-7.06(m,2H),5.93- 5.65(m,2H),5.22-4.91(m,1H),4.48-4.15(m,1H),3.67-3.58(m,1H),2.89-2.78(m,1H),1.70-1.53(m,3H),0.87-0.59(m,3H);

m/z (ESI): [M+H] ⁺ = 515.2.

Example 29

Preparation of Example 29

Under a nitrogen atmosphere, triethylamine (139.4 μL, 1.00 mmol) was added to 0.5 mL of dichloromethane containing compound 24-3 (100 mg, 0.25 mmol), followed by the addition of 4-aminopyridine-2-sulfonamide (45.17 mg, 0.26 mmol), and the mixture was stirred at 26°C for 2 hours. After completion of the reaction, the mixture was quenched with 20 mL of water and extracted with dichloromethane (10 mL × 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product, which was then purified by reverse phase preparative chromatography (Column Waters Xbridge 150*25 mm*5 μm; mobile phase: water (NH ₃ H ₂ O)-ACN; B%: 38%-68%, 12 min) to obtain Example 29.

¹ H NMR (400MHz, DMSO-d ₆ ): δ10.52(s,1H),8.09-8.31(m,1H),7.75(dt,J＝7.6,2.0Hz,1H),7.44-7.61(m,2H),7.36(s,2H),7.08-7.28(m,2H) ,5.10(d,J＝10.4Hz,1H),4.34(dd,J＝10.32,7.44Hz,1H),3.64(s,1H),2.85(s,1H),1.62(s,3H),0.63-0.83(m,3H);

m/z (ESI): [M+H] ⁺ = 535.2.

Examples 30 and 31

Preparation of compound 30-1

Under a nitrogen atmosphere, a solution of compound 24-3 (300 mg, 0.75 mmol) in 2 mL of dichloromethane was added dropwise to a solution of 3-(methylthio)aniline (35.16 mg, 0.25 mmol) and triethylamine (0.3 mL, 2.26 mmol) in 2 mL of dichloromethane. The reaction mixture was stirred at 26°C for 2 hours. After completion of the reaction, the mixture was diluted with 50 mL of water, and the aqueous phase was extracted with dichloromethane (20 mL x 2). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product, which was then purified by silica gel column chromatography (petroleum ether/ethyl acetate = 20/1 to 5/1) to obtain compound 30-1.

¹ H NMR (400MHz, CDCl ₃ ): δ8.30(s,1H),7.57-7.51(m,1H),7.26-7.20(m,2H),7.20-7.14(m,1H),7.03-6.94(m,2H),5.00(d,J＝11.2Hz, 1H),4.28(dd,J=7.6,11.2Hz,1H),2.88-2.76(m,1H),2.47(s,3H),2.37(s,1H),1.70(s,3H),0.84-0.76(m,3H);

m/z(ESI):[M+H] ⁺ =502.1.

Preparation of Examples 30 and 31

Under a nitrogen atmosphere, diacetoxyiodobenzene (407 mg, 1.26 mmol) was added to a solution of compound 30-1 (300 mg, 0.60 mmol) and ammonium acetate (73.8 mg, 0.96 mmol) in 5 mL of methanol. The reaction mixture was stirred at 26°C for 1 hour. After completion of the reaction, the low-boiling solvent was removed by concentration under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography (petroleum ether/ethyl acetate = 1/1 to 0/1) and reverse phase preparative chromatography (0.1% FA/ACN, 50% to 60% over 5 min) to afford a mixture of Examples 30 and 31. The mixture was then separated by SFC (column: DAICEL CHIRALPAK AD (250 mm × 30 mm, 10 μm), CO₂ _{- EtOH} (0.1% _NH₃H₂O ), 30% to 30% over 2.7 min, 60 mL/min) to afford two distinct isomers.

Example 30 (retention time = 0.915 minutes):

¹ H NMR (400MHz, DMSO-d ₆ ): δ10.53(s,1H),8.24(s,1H),7.85(d,J＝8.4Hz,1H),7.63(d,J＝8.0Hz,1H),7.58-7.49(m,1H),7.27-7.14(m,2H),5.09(d,J＝10.4Hz, 1H),4.33(dd,J＝7.6,10.4Hz,1H),4.22-4.15(m,1H),3.64(s,1H),3.02(s,3H),2.92-2.77(m,1H),1.61(s,3H),0.74(d,J＝6.0Hz,3H);

m/z (ESI): [M+H] ⁺ = 533.3.

Example 31 (retention time = 1.165 minutes):

¹ H NMR (400MHz, DMSO-d ₆ ): δ10.54(s,1H),8.24(t,J＝1.8Hz,1H),7.88-7.82(m,1H),7.63(d,J＝8.0Hz,1H),7.58-7.49(m,1H),7.30-7.11(m,2H),5.09(d,J＝10.4Hz ,1H),4.33(dd,J＝7.6,10.4Hz,1H),4.19(s,1H),3.64(s,1H),3.02(d,J＝0.8Hz,3H),2.90-2.77(m,1H),1.61(s,3H),0.74(d,J＝6.0Hz,3H);

m/z (ESI): [M+H] ⁺ = 533.2.

Example 32

Preparation of compound 32-2

Under nitrogen, tert-butyldimethylsilyl chloride (865 mg, 5.74 mmol) was added to a solution of compound 32-1 (300 mg, 2.87 mmol) and imidazole (586 mg, 8.61 mmol) in 5 mL of N,N-dimethylformamide. The reaction mixture was allowed to react at 20°C for 3 hours. After completion, the reaction was quenched with 50 mL of water and extracted with ethyl acetate (25 mL x 2). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product, which was then purified by column chromatography (eluted with petroleum ether) to afford compound 32-2.

¹ H NMR (400MHz, CDCl ₃ ): δ4.37 (t, J = 2.0 Hz, 2H), 4.18 (t, J = 2.0 Hz, 2H), 0.92 (s, 9H), 0.13 (s, 6H).

Preparation of compound 32-3

Under nitrogen atmosphere, a solution of compound 32-2 (28.6 mg, 0.13 mmol) in N,N-dimethylformamide (1 mL) was slowly added to a solution of compound 1-18 (60 mg, 0.13 mmol) and potassium carbonate (90.3 mg, 0.65 mmol) in 1 mL of N,N-dimethylformamide. The reaction mixture was heated to 60°C and stirred for 2 hours. After completion of the reaction, the reaction mixture was cooled and filtered, and the filtrate was separated by reverse phase preparative separation to obtain compound 32-3.

Preparation of Example 32

Compound 32-3 (25 mg, 0.04 mmol) was dissolved in methanolic hydrochloric acid (2 mL, 2 M) and stirred at 25°C for 0.5 hour. After completion of the reaction, the mixture was concentrated, diluted with 2 mL of methanol, and adjusted to pH 8 with aqueous ammonia. The mixture was separated and purified using reverse phase preparative chromatography (column: Waters Xbridge 150×25 mm×5 μm, water (NH ₄ HCO ₃ )-ACN, 38%-68% over 9 min) to obtain Example 32.

¹ H NMR (400MHz, DMSO-d ₆ ): δ10.73(s,1H),8.49(d,J＝5.6Hz,1H),8.26(d,J＝2.0Hz,1H),8.08(d,J＝2.0Hz,1 H),7.84(dd,J＝2.0,5.6Hz,1H),7.63(d,J＝1.6Hz,1H),7.28-7.14(m,2H),5.21(t, J＝6.0Hz,1H),5.11(d,J＝10.4Hz,1H),4.94(d,J＝1.6Hz,2H),4.34(dd,J＝7.6,10.4 Hz,1H),4.10-4.03(m,2H),2.91-2.79(m,1H),1.62(s,3H),0.74(d,J＝6.4Hz,3H);

m/z (ESI): [M+H] ⁺ = 528.2.

Example 33

Preparation of Example 33

Under nitrogen atmosphere, [1,1-bis(diphenylphosphino)ferrocene]dichloropalladium dichloromethane (13.81 mg, 0.02 mmol), cuprous iodide (6.48 mg, 0.03 mmol), triethylamine (117.5 μL, 0.85 mmol) and compound 33-1 (142.23 mg, 1.69 mmol) were added to a 2 mL N,N-dimethylformamide solution of compound 3-1 (100 mg, 0.17 mmol), and the reaction solution was stirred at 100 ° C for 16 hours. After completion of the reaction, the reaction solution was cooled to room temperature, quenched with 40 mL of water, and extracted with ethyl acetate (30 mL × 3). The combined organic phases were washed with 50 mL of saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product, which was then purified by reverse phase preparative chromatography (Column: Waters Xbridge 150 × 25 mm × 5 μm; Condition: water (NH ₄ HCO ₃ )-ACN; Begin B: 38, End B: 68; Gradient Time (min): 10; 100% B Hold Time (min): 4, Flow Rate (ml/min): 25) to obtain Example 33.

¹ H NMR (400 MHz, DMSO-d ₆ ): δ10.75(s,1H),8.49(d,J＝5.4Hz,1H),8.29(d,J＝2.0Hz,1H),8.06(d,J＝1.6Hz ,1H),7.84(dd,J＝2.0,5.4Hz,1H),7.62(d,J＝1.6Hz,1H),7.53-7.45(m,1H),7.2 8(dd,J＝4.0,8.8Hz,1H),5.69(s,1H),5.22(d,J＝10.2Hz,1H),4.36(dd,J＝7.6,1 0.2Hz,1H),2.96(t,J=7.6Hz,1H),1.64(s,3H),1.51(s,6H),0.75-0.68(m,3H);

m/z (ESI): [M-17] ⁺ = 508.2.

Example 34

Preparation of compound 34-2

Under a nitrogen atmosphere, benzoyl chloride (322.7 μL, 2.78 mmol) and triethylamine (478.0 μL, 3.44 mmol) were added to a solution of compound 34-1 (500 mg, 2.65 mmol) in 10 mL of dichloromethane. The reaction was stirred at 25°C for 12 hours. After completion, the reaction was quenched with 30 mL of 1N sodium hydroxide solution. The aqueous phase was extracted with dichloromethane (20 mL x 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product, which was then separated by column chromatography (petroleum ether/ethyl acetate = 10/1) to obtain compound 34-2.

¹ H NMR (400MHz, CDCl ₃ ): δ8.79 (s, 2H), 8.14 (dd, J = 1.2, 8.0Hz, 2H), 7.63-7.56 (m, 1H), 7.50-7.44 (m, 2H), 5.53 (s, 2H);

m/z (ESI): [M+H] ⁺ = 294.4.

Preparation of compound 34-3

Under a nitrogen atmosphere, cesium carbonate (607.20 mg, 1.86 mmol), (±)-2,2-bis(diphenylphosphino)-1,1-binaphthyl (178.53 mg, 0.29 mmol), palladium acetate (32.18 mg, 0.14 mmol), and benzophenone imine (312 mg, 1.72 mmol) were added to a 10 mL toluene solution of compound 34-2 (440 mg, 1.43 mmol). The reaction solution was stirred at 120°C for 12 hours. After completion of the reaction, the reaction solution was cooled to room temperature, quenched with 60 mL of water, and extracted with ethyl acetate (30 mL x 3). The combined organic phases were washed with 50 mL of saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product, which was then separated by column chromatography (petroleum ether/ethyl acetate = 5/1) to obtain compound 34-3.

¹ H NMR (400MHz, CDCl ₃ ): δ8.18(s,2H),8.14-8.10(m,2H),7.81-7.74(m,2H),7.58-7.55(m,1H),7.52(d,J＝7.2 Hz,1H),7.48-7.42(m,4H),7.38-7.32(m,3H),7.13(dd,J=2.0,7.2Hz,2H),5.47(s,2H);

m/z(ESI):[M+H] ⁺ =394.1.

Preparation of compound 34-4

Under a nitrogen atmosphere, hydroxylamine hydrochloride (250.83 mg, 3.61 mmol) and sodium acetate (434.27 mg, 5.29 mmol) were added to a 10 mL methanol solution of compound 34-3 (600 mg, 1.20 mmol). The reaction mixture was stirred at 25°C for 7 hours. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to obtain the crude product, which was then separated by column chromatography (petroleum ether/ethyl acetate = 5/1 to 1/1) to obtain compound 34-4.

¹ H NMR (400MHz, CDCl ₃ ): δ8.22 (s, 2H), 8.12 (d, J = 7.2Hz, 2H), 7.60-7.54 (m, 1H), 7.48-7.41 (m, 2H), 5.47 (s, 2H);

m/z(ESI):[M+23] ⁺ =252.1.

Preparation of compound 34-5

Under nitrogen atmosphere, 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (120.62 mg, 0.32 mmol), N,N-diisopropylethylamine (69.9 μL, 0.42 mmol) and compound 34-4 (64.93 mg, 0.25 mmol) were added to a solution of compound 21-4 (80 mg, 0.21 mmol) in 2 mL of N,N-dimethylformamide. The reaction solution was stirred at 25°C for 16 hours. After completion of the reaction, the mixture was quenched with 40 mL of water and extracted with ethyl acetate (30 mL × 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product, which was then separated by column chromatography (petroleum ether/ethyl acetate = 5/1 to 3/1) to obtain compound 34-5.

¹ H NMR (400 MHz, CDCl ₃ ): δ8.97(s,2H),8.38(s,1H),8.16-8.09(m,2H),7.63-7.55(m,1H),7.50-7 .41(m,2H),7.17-7.09(m,1H),7.04-6.94(m,1H),5.53(s,2H),5.04(d,J＝11 .2Hz,1H),4.93-4.87(m,1H),4.83-4.78(m,1H),4.26(dd,J＝7.6,11.2Hz,1H ),2.89-2.81(m,1H),2.44(t,J＝2.4Hz,1H),1.70(s,3H),0.84-0.79(m,3H);

m/z(ESI):[M+H] ⁺ =590.2.

Preparation of Example 34

Under a nitrogen atmosphere, lithium hydroxide (38.05 mg, 0.91 mmol) was added to a 3 mL methanol solution of compound 34-5 (110 mg, 0.18 mmol), and the reaction mixture was stirred at 25°C for 1 hour. After completion of the reaction, the mixture was concentrated under reduced pressure to obtain a crude product, which was then purified by reverse phase preparative purification (Column: Waters Xbridge 150×25 mm×5 μm; Condition: water (NH ₄ HCO ₃ )-ACN; Begin B: 23, End B: 53; Gradient Time (min): 10; 100% B Hold Time (min): 4, Flow Rate (ml/min): 25) to obtain Example 34.

1H NMR (400MHz, DMSO-d ₆ ): δ10.46(s,1H),8.98(s,2H),7.27-7.16(m,2H),5.26(t,J＝6.0Hz,1H),5.15(d,J＝10.4Hz,1H),4.98-4.85(m,2H),4.55(d ,J＝6.0Hz,2H),4.32(dd,J＝7.2,10.2Hz,1H),3.67(t,J＝2.0Hz,1H),2.89-2.78(m,1H),1.62(s,3H),0.74(d,J＝6.0Hz,3H);

m/z(ESI):[M+H] ⁺ =486.1.

Example 35

Preparation of compound 35-2

Under nitrogen atmosphere, triethylamine (94.1 mg, 0.93 mmol), tert-butyldimethyl(2-propynyloxy)silane (317 mg, 1.86 mmol), (diphenylphosphino)ferrocenepalladium chloride (13.6 mg, 0.02 mmol), and cuprous iodide (7.08 mg, 0.04 mmol) were added sequentially to a solution of compound 3-1 (110 mg, 0.19 mmol) in 2 mL of N,N-dimethylformamide. The reaction mixture was stirred at 80°C for 48 hours. After completion of the reaction, the reaction mixture was cooled to room temperature, diluted with 30 mL of water, and extracted with ethyl acetate (15 mL × 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product, which was then isolated and purified by reverse phase preparative (0.1% FA/ACN, 80% to 90% over 5 min) to obtain compound 35-2.

m/z(ESI):[MH] ⁺ ＝612.2

Preparation of Example 35

Under nitrogen atmosphere, compound 35-2 (14 mg, 0.02 mmol) was added to a 2 M solution of methanolic hydrochloric acid (2 mL), and the reaction solution was stirred at 25°C for 0.5 h. After completion of the reaction, the reaction solution was concentrated to dryness to obtain the crude product, which was separated and purified by reverse phase preparation (column: Phenomenex luna C18 150×25mm×10um, 35% to 65% over 9 min) and (column: Waters Xbridge 150×25mm×5um, 35% to 65% over 11 min) to obtain Example 35.

¹ H NMR (400MHz, DMSO-d ₆ ): δ10.78 (br s,1H),8.47(d,J＝5.2Hz,1H),8.27(s,1H),8.08-8.03(m,1H),7.83(d,J ＝3.6Hz,1H),7.60(s,1H),7.52-7.45(m,1H),7.30(dd,J＝4.4,8.5Hz,1H ),5.67-5.45(m,1H),5.20(d,J＝10.4Hz,1H),4.42(s,2H),4.36(dd,J＝7.6,10.3Hz,1H),3.04-2.92(m,1H),1.62(s,3H),0.71(d,J＝6.4Hz,3H);

m/z (ESI): [M+H] ⁺ = 498.2.

Examples 36 and 37

Preparation of compound 36-2

Under a nitrogen atmosphere, compound 36-1 (35.34 mg, 0.25 mmol) was added to a 2 mL dichloromethane solution containing compound 21-5 (100 mg, 0.25 mmol) and triethylamine (105.1 μL, 0.76 mmol). The reaction mixture was allowed to react at 25°C for 1 hour. After completion, 3 mL of water was added to dilute and quench the reaction. The aqueous phase was extracted with dichloromethane (10 mL × 3). The combined organic phases were washed with 5 mL of saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product, which was then isolated and purified by reverse phase preparative chromatography (0.1% formic acid) to obtain compound 36-2.

¹ H NMR (400MHz, CDCl ₃ ): δ8.38-8.29(m,2H),7.51(d,J＝1.6Hz,1H),7.09(dd,J＝1.6,5.6Hz,1H),7.03-6.96(m,1H),4.99(d,J＝11.2Hz,1H),4.92-4.86(m,1H),4.83 -4.77(m,1H),4.25(dd,J＝7.8,11.2Hz,1H),2.88-2.77(m,1H),2.54(s,3H),2.38(t,J＝2.4Hz,1H),1.70(s,3H),0.80(dd,J＝1.6,7.4Hz,3H);

m/z (ESI): [M+H] ⁺ = 501.1.

Preparation of Examples 36 and 37

Under nitrogen atmosphere, iodobenzene diacetate (40.80 mg, 0.13 mmol) and ammonium acetate (7.39 mg, 0.10 mmol) were added sequentially to a 1 mL methanol solution of compound 36-2 (30 mg, 0.06 mmol), and the reaction solution was reacted at 25°C for 18 hours. After completion of the reaction, the reaction solution was concentrated to obtain a crude product, which was purified _by reverse phase preparative purification (Waters Xbridge 150×25 mm×5 um; mobile phase: water ( _NH4HCO3 )-ACN; B%: 43%-63%, 25 min) to obtain a mixture of Example 36 and Example _37. The mixture was separated by SFC (Column DAICEL CHIRALPAK AD (250×50 mm×10 um); Condition: CO2 _- EtOH (0.1% _NH3H2O ); B%: 20-20; Gradient Time (min): 3.6; 100% B Hold Time (min): 0; Flow Rate (ml/min): 120) to obtain two single isomers.

Example 36 (retention time = 1.273 minutes):

¹ H NMR (400MHz, DMSO-d ₆ ): δ10.88(s,1H),8.57(d,J＝5.6Hz,1H),8.36(d,J＝2.0Hz,1H),7.82(dd,J＝2.0,5.6Hz,1H),7.27-7.17(m,2H),5.13(d,J＝10.4Hz,1H),4.9 5-4.89(m,2H),4.37-4.30(m,2H),3.64(t,J＝2.4Hz,1H),3.11(d,J＝0.8Hz,3H),2.84(t,J＝7.2Hz,1H),1.61(s,3H),0.74(d,J＝6.4Hz,3H);

m/z(ESI):[M+H] ⁺ =532.1.

Example 37 (retention time = 1.420 minutes):

¹ H NMR (400MHz, DMSO-d ₆ ): δ10.89(s,1H),8.57(d,J＝5.6Hz,1H),8.35(d,J＝2.0Hz,1H),7.82(dd,J＝2.0,5.6Hz,1H),7.27-7.16(m,2H),5.13(d,J＝10.4Hz,1H),4.9 2(t,J＝2.8Hz,2H),4.39-4.29(m,2H),3.64(t,J＝2.4Hz,1H),3.11(d,J＝0.8Hz,3H),2.87-2.80(m,1H),1.61(s,3H),0.74(d,J＝5.6Hz,3H);

m/z (ESI): [M+H] ⁺ = 532.2.

Example 38

Preparation of Example 38

Under a nitrogen atmosphere, tris(dibenzylideneacetone)dipalladium (7.74 mg, 0.01 mmol), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (4.89 mg, 0.01 mmol), and N,N-diisopropylethylamine (41.9 μL, 0.25 mmol) were added sequentially to compound 3-1 (50 mg, 0.08 mmol) and dimethylphosphine oxide (13.20 mg, 0.17 mmol) in 2 mL of dioxane. The reaction mixture was heated to 100°C and stirred for 15 hours. After completion of the reaction, the reaction mixture was cooled to room temperature and filtered through a short silica gel column (washed with ethyl acetate). The filtrate was concentrated under reduced pressure to obtain the crude product, which was then purified by reverse phase preparative chromatography (Waters Xbridge 150×25 mm×5 μm; mobile phase: water (NH ₄ HCO ₃ )-ACN; B%: 23%-53%, 25 min) to obtain Example 38.

¹ H NMR (400MHz, DMSO-d ₆ ): δ10.60(s,1H),8.48(d,J＝5.5Hz,1H),8.28(d,J＝2.1Hz,1H),8.06(d,J＝2.1Hz,1H),7.83(dd,J＝2.2,5.6Hz,1H),7.72-7.59(m,2H),7.46-7. 39(m,1H),5.48(dd,J＝7.1,10.8Hz,1H),5.15(d,J＝10.8Hz,1H),2.90(t,J＝7.4Hz,1H),1.85-1.76(m,6H),1.58(s,3H),0.76(d,J＝6.5Hz,3H);

m/z(ESI):[M+H] ⁺ =520.2.

Example 39

Preparation of Example 39

Under a nitrogen atmosphere, triethylamine (139.4 μL, 1.00 mmol) was added to 0.5 mL of dichloromethane containing compound 24-3 (100 mg, 0.25 mmol), followed by the addition of 4-aminopyridine-2-sulfonamide (45.17 mg, 0.26 mmol), and the mixture was stirred at 26°C for 2 hours. After completion of the reaction, the mixture was quenched with 20 mL of water and extracted with dichloromethane (10 mL × 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product, which was then purified by reverse phase preparative chromatography (Column Waters Xbridge 150*25 mm*5 μm; mobile phase: water (NH ₃ H ₂ O)-ACN; B%: 38%-68%, 12 min) to obtain Example 39.

¹ H NMR (400MHz, DMSO-d ₆ ): δ10.52(s,1H),8.09-8.31(m,1H),7.75(dt,J＝7.6,2.0Hz,1H),7.44-7.61(m,2H),7.36(s,2H),7.08-7.28(m,2H) ,5.10(d,J＝10.4Hz,1H),4.34(dd,J＝10.32,7.44Hz,1H),3.64(s,1H),2.85(s,1H),1.62(s,3H),0.63-0.83(m,3H);

m/z (ESI): [M+H] ⁺ = 535.2.

Example 40

Preparation of compound 40-2

Under a nitrogen atmosphere, triethylamine (57.5 μL, 0.41 mmol) and compound 40-1 (43.28 mg, 0.21 mmol) were added to a 1 mL dichloromethane solution of compound 24-3 (55.0 mg, 0.14 mmol). The reaction mixture was allowed to react at 25°C for 1 hour. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to obtain the crude product, which was then isolated and purified by silica gel column chromatography (petroleum ether/ethyl acetate = 3/1) to obtain compound 40-2.

¹ H NMR (400MHz, CDCl ₃ ): δ8.51(s,1H),7.82-7.77(m,3H),7.20-7.12(m,1H),7.03-6.94(m,1H),5.02(d,J＝11.2Hz,1H),4.26(dd,J ＝7.6,11.2Hz,1H),3.90(s,3H),3.89(s,3H),2.90-2.78(m,1H),2.37(s,1H),1.71(s,3H),0.85-0.78(m,3H);

m/z (ESI): [M+H] ⁺ = 572.2.

Preparation of Example 40

Under nitrogen atmosphere, compound 40-2 (50.0 mg, 0.08 mmol) was dissolved in 5 mL of ammonia methanol (7 M) solution, and the reaction mixture was reacted at 60°C for 17 hours. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to obtain the crude product, which was then isolated and purified via reverse phase preparative chromatography (Column Phenomenex luna C18 150 × 25 mm × 10 μm; mobile phase: water (FA)-ACN; B%: 43%-73%, 11 min) to obtain Example 40.

¹ H NMR (400MHz, METHANOL-d ₄ ): δ8.54(s,1H),7.82(d,J＝2.0Hz,1H),7.73(dd,J＝2.0,8.4Hz,1H),7.59(d,J＝8.4Hz,1H),7.23-7.13(m,1H),7.09-6.99(m,1 H),5.08(d,J＝10.8Hz,1H),4.43(dd,J＝8.0,10.2Hz,1H),2.93(s,1H),2.90-2.84(m,1H),1.68(s,3H),0.83(d,J＝5.6Hz,3H);

m/z (ESI): [M+H] ⁺ = 542.2.

Example 41

Preparation of Example 41

Under nitrogen atmosphere, m-chloroperbenzoic acid (55.0 mg, 0.32 mmol) was added to a solution of compound 24-5 (80 mg, 0.16 mmol) in 3 mL of dichloromethane at 0°C, and the reaction solution was stirred at 0°C for 2 hours. After completion of the reaction, the reaction solution was slowly added to 50 mL of saturated aqueous sodium sulfite solution for quenching. The aqueous phase was negative when measured with starch potassium iodide paper, and then extracted with dichloromethane (20 mL×2). The combined organic phases were washed with 20 mL of saturated sodium bicarbonate, _dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product, which was separated and purified by reverse phase preparation (column: Waters Xbridge C18150*25mm*5um; mobile phase: [A: _H2O ( _10mM NH4HCO3); B: ACN]; B%: 48.00%-78.00%, 15.00min; flow rate: 25.00ml/min) to obtain Example 41.

¹ H NMR (400MHz, DMSO-d ₆ ): δ10.90(s,1H),8.63(d,J＝5.6Hz,1H),8.36(d,J＝2.0Hz,1H),7.91(dd,J＝2.0,5.6Hz,1H),7.30-7.15(m,2H),5.14(d,J＝1 0.4Hz,1H),4.33(dd,J＝7.6,10.4Hz,1H),3.62(s,1H),3.24(s,3H),2.90-2.77(m,1H),1.62(s,3H),0.74(d,J＝6.4Hz,3H);

m/z (ESI): [M+H] ⁺ = 535.2.

Example 42

Preparation of compound 42-2

Under nitrogen atmosphere, dimethylphosphine oxide (676.69 mg, 8.67 mmol), potassium phosphate (3.07 g, 14.45 mmol), tris(dibenzylideneacetone)dipalladium (317.58 mg, 0.35 mmol), and 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (401.34 mg, 0.69 mmol) were added to a solution of compound 42-1 (1.0 g, 5.78 mmol) in 20 mL of N,N-dimethylformamide. The reaction mixture was heated to 140°C and reacted for 16 hours. After the reaction was completed, the reaction mixture was cooled to room temperature and filtered. The filter cake was washed twice with 10 mL of ethyl acetate, and the filtrate was concentrated under reduced pressure to obtain the crude product, which was purified by silica gel column chromatography (dichloromethane/methanol = 10:1) to obtain compound 42-2.

¹ H NMR (400MHz, DMSO-d ₆ ): δ8.09 (d, J = 5.6 Hz, 1H), 7.10 (dd, J = 2.4, 7.2 Hz, 1H), 6.52 (td, J = 2.0, 5.6 Hz, 1H), 6.34 (s, 2H), 1.56 (s, 3H), 1.53 (s, 3H);

m/z (ESI): [M+H] ⁺ = 171.2.

Preparation of Example 42

Under a nitrogen atmosphere, compound 42-2 (42.93 mg, 0.21 mmol) and triethylamine (57.5 μL, 0.41 mmol) were sequentially added to a 1 mL dichloromethane solution of compound 24-3 (55 mg, 0.14 mmol). The reaction mixture was reacted at 25°C for 1 hour. After completion of the reaction, the mixture was concentrated under reduced pressure to obtain a crude product, which was then purified by reverse phase preparative chromatography (Waters Xbridge C18 column, 150×25 mm×5 μm, aqueous phase-organic phase H ₂ O (10 mM NH ₄ HCO ₃ )-ACN, B%: 33%-63%, 15 min) to obtain Example 42.

¹ H NMR (400MHz, DMSO-d ₆ ): δ10.74(s,1H),8.61(d,J＝5.6Hz,1H),8.21(dd,J＝2.0,6.4Hz,1H),7.77(td,J＝2.0,5.6Hz,1H),7.27-7.13(m,2H),5.12 (d,J＝10.6Hz,1H),4.33(dd,J＝7.2,10.2Hz,1H),3.65(s,1H),2.91-2.77(m,1H),1.70-1.51(m,9H),0.74(d,J＝6.0Hz,3H);

m/z (ESI): [M+H] ⁺ = 533.2.

Example 43

Preparation of compound 43-3

Under a nitrogen atmosphere, compound 43-2 (2.69 g, 20.34 mmol), triphenylphosphine (5.34 g, 20.34 mmol), and diisopropyl azodicarboxylate (4.0 mL, 20.34 mmol) were added to a 30 mL toluene solution of compound 43-1 (1.9 g, 13.56 mmol). The reaction mixture was heated to 80°C and allowed to react for 3 hours. After completion of the reaction, the reaction mixture was cooled to room temperature and concentrated under reduced pressure to obtain the crude product, which was then separated by column chromatography (petroleum ether/ethyl acetate = 10:1) to obtain compound 43-3.

¹ H NMR (400MHz, CDCl ₃ ): δ8.39(d,J＝5.6Hz,1H),7.61(dd,J＝2.0,5.6Hz,1H),7.53(d,J＝2.0Hz,1H),4.57-4.39( m,3H),4.17(dd,J＝6.4,8.4Hz,1H),3.88(dd,J＝6.0,8.4Hz,1H),1.47(s,3H),1.41(s,3H).

Preparation of compound 43-4

Under a nitrogen atmosphere, iron powder (2.42 g, 43.42 mmol) and ammonium chloride (2.32 g, 43.42 mmol) were added to compound 43-3 (2.3 g, 8.68 mmol) dissolved in 20 mL of ethanol and 4 mL of aqueous solution. The reaction mixture was heated to 80°C and allowed to react for 2 hours. After completion of the reaction, the reaction mixture was cooled to room temperature, diluted with 40 mL of ethyl acetate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product, which was then isolated by column chromatography (petroleum ether/ethyl acetate = 10/1 to 1/1) to obtain compound 43-4.

¹ H NMR (400MHz, DMSO-d ₆ ): δ7.59(d,J＝6.0Hz,1H),6.17(dd,J＝2.0,6.0Hz,1H),5.93(s,2H),5.81(d,J＝1.6Hz,1H),4.37-4.29(m,1 H),4.20-4.09(m,2H),4.03(dd,J＝6.4,8.0Hz,1H),3.69(dd,J＝6.4,8.0Hz,1H),1.33(s,3H),1.28(s,3H);

m/z (ESI): [M+H] ⁺ = 225.2.

Preparation of compound 43-5

Under a nitrogen atmosphere, compound 43-5 (86.97 mg, 0.38 mmol) and triethylamine (104.6 μL, 0.75 mmol) were added to a solution of compound 24-3 (100 mg, 0.25 mmol) in 1 mL of dichloromethane. The reaction mixture was allowed to react at 25°C for 1 hour. After completion, the reaction was quenched with 20 mL of water and extracted with dichloromethane (20 mL x 2). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product, which was then separated by silica gel column chromatography (petroleum ether/ethyl acetate = 10/1 to 3/1) to obtain compound 43-5.

¹ H NMR (400 MHz, CDCl ₃ ): δ8.37(s,1H),8.03(d,J＝6.4Hz,1H),7.17-7.10(m,1H),7.06-6.95(m,3H),4 .98(d,J＝11.2Hz,1H),4.49-4.42(m,1H),4.37-4.30(m,2H),4.25(dd,J＝8.0,11 .3Hz,1H),4.12(dd,J＝6.0,8.4Hz,1H),3.85(dd,J＝6.0,8.4Hz,1H),2.89-2.77 (m,1H),2.37(s,1H),1.69(s,3H),1.45(s,3H),1.39(s,3H),0.83-0.78(m,3H);

m/z (ESI): [M+H] ⁺ = 587.2.

Preparation of Example 43

Under a nitrogen atmosphere, trifluoroacetic acid (0.2 mL, 2.69 mmol) was added to a 2 mL dichloromethane solution of compound 43-5 (50 mg, 0.08 mmol), and the reaction mixture was allowed to react at 26°C for 1 hour. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to obtain the crude product, which was then purified via reverse phase preparative chromatography (column: Waters Xbridge C18 150*25 mm*5 μm; mobile phase: [A: H ₂ O (10 mM NH ₄ HCO ₃ ); B: ACN]; B%: 40.00%-70.00%, 15.00 min; flow rate: 25.00 ml/min) to obtain Example 43.

¹ H NMR (400MHz, DMSO-d ₆ ): δ10.49(s,1H),8.01(d,J＝5.6Hz,1H),7.26-7.19(m,1H),7.17-7.10(m,3H),5.08(d,J＝ 10.4Hz,1H),4.87(d,J＝5.2Hz,1H),4.60(t,J＝5.6Hz,1H),4.32(dd,J＝7.6,10.0Hz,1H),4 .22(dd,J＝4.4,10.8Hz,1H),4.09(dd,J＝6.0,10.8Hz,1H),3.75(qd,J＝5.6,10.8Hz,1H),3 .63(s,1H),3.40(t,J＝5.6Hz,2H),2.90-2.77(m,1H),1.59(s,3H),0.73(d,J＝6.0Hz,3H);

m/z (ESI): [M+H] ⁺ = 547.2.

Example 44

Preparation of Example 44

Under a nitrogen atmosphere, compound 24-3 (424 mg, 1.06 mmol) was dissolved in dichloromethane (6 mL), and triethylamine (0.5 mL, 3.6 mmol) was added, followed by compound 44-1 (127 mg, 1.06 mmol). The reaction solution was stirred at room temperature for 3 hours. After completion of the reaction, the reaction solution was quenched by adding 20 mL of saturated aqueous sodium bicarbonate solution. The aqueous phase was extracted with dichloromethane (3 × 20 mL), and the combined organic phases were washed with saturated brine (1 × 30 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was isolated and purified by column chromatography (petroleum ether/ethyl acetate = 10/1 to 1/1) to obtain Example 44.

LC-MS (ESI): [M+H] ⁺ = 482.1.

Example 45

Preparation of Example 45

Compound 45-1 (17.95 mg, 0.16 mmol) and triethylamine (87.1 μL, 0.63 mmol) were dissolved in DMF (1 mL) and stirred at room temperature for 5 minutes. A solution of compound 24-3 (50 mg, 0.13 mmol) in DMF (1 mL) was then slowly added dropwise to the mixture, followed by stirring at room temperature for 2 hours. After the reaction was complete, water (10 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (10 mL × 3). The organic phase was washed with saturated sodium chloride aqueous solution (10 mL × 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by thin layer silica gel preparation (dichloromethane:methanol=10:1) and reverse phase preparation (Phenomenex luna C18 150×25mm×10um; flow rate: 25mL/min; gradient: 47%-67% B over 10min; mobile phase A: 0.225% aqueous methanoic acid, mobile phase B: acetonitrile) to obtain Example 45.

¹ H NMR (400MHz, CDCl ₃ ): δ8.42-8.27(m,1H),7.15-6.94(m,3H),6.75-6.62(m,2H),5.06-4.93(m,1H),4.36-4.2 0(m,1H),2.88-2.78(m,1H),2.47-2.40(m,1H),1.36-1.18(m,3H),0.80(d,J＝5.2Hz,3H);

m/z(ESI):[M+H] ⁺ =473.1.

Example 46

Preparation of Example 46

Under nitrogen atmosphere, sodium methoxide in methanol (0.01 mL, 0.05 mmol) was added to Example 44 (100 mg, 0.21 mmol) in 5 mL of methanol. The reaction was stirred at room temperature for 2 hours, followed by the addition of ammonium chloride (23 mg, 0.42 mmol), and the temperature was raised to 40°C and stirring continued for 12 hours. The reaction was concentrated, and the crude product was purified via reverse elution (Phenomenex luna C18 150×25 mm×10 μm; flow rate: 25 mL/min; gradient: 23%-53% B over 10 min; mobile phase A: 0.225% aqueous methanoic acid, mobile phase B: acetonitrile) to afford Example 46.

¹ H NMR (400MHz, DMSO-d ₆ ): δ8.61(d,J＝5.6Hz,1H),8.47-8.44(m,2H),7.89-7.87(m,1H),7.23-7.21(m,2H),5.19(d,J＝10 .8Hz,1H),4.38-4.33(m,1H),3.66(s,1H),2.89-2.81(m,1H),1.62(s,3H),0.75(d,J＝6.0Hz,3H);

m/z(ESI):[M+H] ⁺ =499.1.

Example 47

Preparation of Example 47

Under nitrogen atmosphere, Example 44 (100 mg, 0.21 mmol) was dissolved in 2 mL of 1,4-dioxane, and triethylamine (0.058 mL, 0.42 mmol) and dimethylamine hydrochloride (33.87 mg, 0.42 mmol) were added sequentially. The reaction solution was heated to 90°C and stirred for 16 hours. The reaction solution was concentrated, and the crude product was purified via reverse elution (Phenomenex luna C18 150×25 mm×10 μm; flow rate: 25 mL/min; gradient: 24%-54% B over 10 min; mobile phase A: 0.225% aqueous methanoic acid, mobile phase B: acetonitrile) to give Example 47.

¹ H NMR (400MHz, DMSO-d ₆ ): δ11.55-11.48(m,1H),8.57(d,J＝5.6Hz,1H),8.44(s,1H),8.04(d,J＝1.6Hz,1H),7.78-7.76(m,1H),7.30-7.26(m,1H),7.23-7.17 (m,1H),5.26(d,J＝10.4Hz,1H),4.37-4.33(m,1H),3.66(s,1H),3.02(s,6H),2.87-2.78(m,1H),1.61(s,3H),0.75(d,J＝6.4Hz,3H);

m/z (ESI): [M+H] ⁺ = 527.2.

Example 48

Preparation of compound 48-1

Compound 12-2 (500 mg, 3.85 mmol), imidazole (800 mg, 11.76 mmol), and tert-butyldiphenylsilyl chloride (2.4 g, 8.73 mmol) were dissolved in anhydrous DCM (30 mL) and stirred at room temperature for 12 hours. The reaction mixture was concentrated to obtain a crude product, which was purified by silica gel column chromatography (eluted with petroleum ether) to obtain compound 48-1.

Preparation of compound 48-2

Compound 48-1 (900 mg, 2.44 mmol) and potassium carbonate (1.1 g, 7.96 mmol) were dissolved in DMF (12 mL) and stirred at room temperature for 24 hours. The reaction mixture was added with ethyl acetate (100 mL), washed once with water (30 mL), and once with saturated brine (30 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and dried. Compound 48-2 was then purified by silica gel column chromatography.

¹ H NMR (400MHz, CDCl3): δ7.56-7.59(m,1H), 7.26-7.50(m,1H), 2.24(s,1H), 1.12-1.34(m,9H).

Preparation of compound 48-3

Under a nitrogen atmosphere, compound 48-2 (370 mg, 1.25 mmol) was dissolved in THF (1 mL) at -50°C. A solution of n-butyllithium in n-hexane (1.0 mL, 2.50 mmol) was slowly added, and the mixture was stirred for 0.5 hours. A solution of deuterated iodomethane (370 mg, 2.55 mmol) in THF (1 mL) was added, and the mixture was stirred for 1 hour. The reaction mixture was quenched by pouring it into a saturated aqueous solution of ammonium chloride (100 mL), and the mixture was extracted with ethyl acetate (100 mL x 3). The organic phase was combined with saturated brine (100 mL), washed once, dried over anhydrous sodium sulfate, filtered, and dried to obtain compound 48-3.

¹ H NMR (400MHz, CDCl ₃ ): δ7.77-7.79(m,1H), 7.44-7.50(m,1H), 1.12-1.34(m,9H).

Preparation of compound 48-4

Compound 48-3 (370 mg, 1.18 mmol) was dissolved in a solution of tetrabutylammonium fluoride in tetrahydrofuran (5 mL, 5.00 mmol) and stirred at room temperature for 0.5 hours. The reaction mixture was diluted with ethyl acetate (100 mL) and then washed once with water (100 mL) and once with saturated brine (100 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and dried to afford compound 48-4.

Preparation of Example 48

Under nitrogen atmosphere, triphenylphosphine (70 mg, 0.11 mmol), compound 1-18 (50 mg, 0.11 mmol) and compound 48-4 (10 mg, 0.13 mmol) were dissolved in THF (1.5 mL). Diisopropyl azodicarboxylate (30 mg, 0.15 mmol) was added under ice-water bath and stirred at room temperature overnight. Ethyl acetate (20 mL) was added to the reaction solution, and the mixture was washed with water (10 mL x 2) and saturated brine (10 mL) once. The organic phase was dried over anhydrous sodium sulfate, filtered, and spin-dried to obtain the crude product, which was purified via reverse PCR (Phenomenex luna C18 150 × 25 mm × 10 μm; flow rate: 25 mL/min; gradient: 48%-78% B over 10 min; mobile phase A: 0.225% aqueous methanoic acid, mobile phase B: acetonitrile) to obtain Example 48.

m/z(ESI):[M+H] ⁺ =517.1.

Example 49

Preparation of Example 49

Example 24 (50 mg, 0.09 mmol) was placed in a 50 mL single-necked bottle. DCM (3 mL) was added at room temperature to replace the argon atmosphere. Trimethyloxonium tetrafluoroborate (7 mg, 0.05 mmol) was added to the solution, and stirred at room temperature for 6 hours. The reaction solution was spin-dried to obtain a crude product, which was purified by thin-layer silica gel chromatography (petroleum ether:ethyl acetate = 1:1) to obtain Example 49.

¹ H NMR (400MHz, DMSO-d ₆ ): δ10.91(s,1H),8.62(d,J＝5.5Hz,1H),8.32(d,J＝1.9Hz,1H),7.86(dd,J＝5.5,2.1Hz,1H),7.32–7.12(m,2H),5.14(d,J＝10.4Hz ,1H),4.34(dd,J＝10.4,7.5Hz,1H),3.63(s,1H),3.16(s,3H),2.93–2.76(m,1H),2.45(s,3H),1.62(s,3H),0.74(d,J＝6.2Hz,3H);

m/z (ESI): [M+H] ⁺ = 548.4.

Example 50

Preparation of compound 50-1

Under a nitrogen atmosphere, a dichloromethane solution of boron tribromide (12.7 mL, 25.4 mmol) was slowly added dropwise to a 60 mL dichloromethane solution of compound 1-12 (4.5 g, 12.7 mmol) at 0°C. The reaction mixture was heated to 25°C and stirred for 1 hour. After completion of the reaction, the reaction mixture was poured into 100 mL of saturated sodium bicarbonate aqueous solution and extracted once with 50 mL of dichloromethane. The organic phase was discarded, and the pH of the aqueous phase was adjusted to 3-4 with 30 mL of dilute hydrochloric acid. The aqueous phase was extracted with dichloromethane (50 mL x 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain compound 50-1.

¹ H NMR (400MHz, DMSO-d ₆ ): δ7.11-6.96(m,1H),6.92-6.70(m,1H),4.97(d,J＝10.4Hz,1H),4.09(dd,J＝10.4,7.6Hz,1H),2.73(s,1H),1.52(s,3H),0.67(d,J＝6.0Hz,3H).

Preparation of compound 50-2

Under a nitrogen atmosphere, 2,2,2-trichloro-1-[(2-methylprop-2-yl)oxy]ethane-1-imine (10.82 g, 49.52 mmol) was added to a solution of compound 50-1 (3.37 g, 9.90 mmol) in 35 mL of dichloromethane. The reaction solution was stirred at 25°C for 1 hour. After completion of the reaction, 20 mL of water was added to quench the reaction. The mixture was extracted with dichloromethane (50 mL x 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product, which was then isolated and purified by silica gel column chromatography (petroleum ether/ethyl acetate = 10/1 to 5/1) to obtain compound 50-2.

¹ H NMR (400MHz, DMSO-d ₆ ): δ7.04(t,J＝6.8Hz,1H),6.91-6.74(m,1H),4.90(d,J＝10.4Hz,1H),4.04(dd,J＝10.8,7. 2Hz,1H),2.70(t,J＝7.6Hz,1H),1.52(d,J＝2.0Hz,5H),1.27(s,9H),0.70(d,J＝5.6Hz,3H).

Preparation of compound 50-3

Under a nitrogen atmosphere, trifluoromethanesulfonic anhydride (4.409 g, 15.63 mmol) was added to a solution of triethylamine (3.163 g, 31.26 mmol) and compound 50-2 (4.130 g, 10.42 mmol) in 40 mL of dichloromethane at 0°C. The reaction mixture was allowed to react at 0°C for 2 hours. After completion of the reaction, the reaction mixture was poured into 100 mL of water, and the aqueous phase was extracted with dichloromethane (50 mL x 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product, which was then isolated and purified by silica gel column chromatography (petroleum ether/ethyl acetate = 5/1 to 2/1) to obtain compound 50-3.

¹ H NMR (400MHz, CDCl ₃ ): δ7.35-7.13(m,2H),4.79(d,J＝9.6Hz,1H),4.23-4.03(m,1H),2.78(t,J＝ 7.6Hz,1H),1.63(s,3H),1.35(s,9H),1.23-1.30(m,2H),0.90-0.78(m,3H).

Preparation of compound 50-4

Under nitrogen atmosphere, potassium vinyl fluoroborate (2.23 g, 16.65 mmol), cesium carbonate (5.426 g, 16.65 mmol) and RuPhos Pd G3 (696 mg, 0.83 mmol) were added sequentially to a solution of compound 50-3 (4.4 g, 8.33 mmol) in 9 mL of water and 45 mL of toluene. The reaction solution was heated to 85°C and reacted for 5 hours. After the reaction was completed, the reaction solution was cooled to room temperature and diluted with 50 mL of water. The aqueous phase was extracted with ethyl acetate (50 mL × 3). The combined organic phases were dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to obtain the crude product, which was separated and purified by silica gel column chromatography (petroleum ether/ethyl acetate = 20/1-5/1) to obtain compound 50-4.

¹ H NMR (400MHz, CDCl ₃ ): δ7.15-6.98(m,2H),6.56(dd,J＝17.6,11.6Hz,1H),5.84-5.46(m,2H),4.89-4.75(m,2H),4. 06-3.94(m,1H),2.58(s,1H),1.56(s,3H),1.31(s,9H),1.29-1.27(m,1H).0.81-0.72(m,3H).

Preparation of compound 50-5

Ozone was introduced into a 50 mL dichloromethane solution containing compound 50-4 (3.287 g, 8.09 mmol). After the reaction solution turned blue, nitrogen was introduced until the solution returned to yellow. Triphenylphosphine (3.18 g, 12.13 mmol) was added, and the reaction solution was slowly heated to 25°C and allowed to react for 12 hours. After completion of the reaction, the reaction solution was evaporated to dryness under reduced pressure to obtain the crude product, which was then isolated and purified by silica gel column chromatography (petroleum ether/ethyl acetate = 20/1 to 10/1) to obtain compound 50-5.

¹ H NMR (400MHz, CDCl ₃ ): δ10.52(s,1H),7.47-7.38(m,1H),7.27(s,2H),4.88(d,J＝10.6Hz,1H),4 .76-4.63(m,1H),2.87(s,1H),1.68(s,3H),1.32(s,9H),0.78-0.69(m,3H).

Preparation of compound 50-6

Under a nitrogen atmosphere, sodium borohydride (92.64 mg, 2.45 mmol) was slowly added to a solution of compound 50-5 (0.5 g, 1.22 mmol) in 6 mL of tetrahydrofuran at 0°C. The reaction mixture was warmed to 25°C and stirred for 1 hour. After completion, the reaction was quenched by the addition of 10 mL of saturated aqueous ammonium chloride. The aqueous phase was extracted with ethyl acetate (10 mL x 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product, which was then isolated and purified by silica gel column chromatography (petroleum ether/ethyl acetate = 10/1 to 5/1) to obtain compound 50-6.

Preparation of compound 50-7

Under a nitrogen atmosphere, sodium hydride (102 mg, 2.56 mmol, 60% content) was slowly added to a solution of compound 50-6 (350 mg, 0.85 mmol) in 6 mL of tetrahydrofuran at 0°C. After stirring for 10 minutes, (3-bromoprop-1-ynyl)trimethylsilane (244 mg, 1.28 mmol) was added. The reaction solution was warmed to 25°C and stirred for 6 hours. After completion, the reaction was quenched by the addition of 5 mL of saturated aqueous ammonium chloride solution and adjusted to pH 3-4 with 0.5 M hydrochloric acid. The aqueous phase was extracted with dichloromethane (10 mL x 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product, which was then isolated and purified by silica gel column chromatography (petroleum ether/ethyl acetate = 10/1 to 1/1) to obtain compound 50-7.

Preparation of compound 50-8

Under nitrogen atmosphere, N,N-dimethylformamide (2.3 μL, 0.03 mmol) and oxalyl chloride (114.52 mg, 0.90 mmol) were added sequentially to 4 mL of dichloromethane solvent containing compound 50-7 (118 mg, 0.30 mmol). The reaction solution was reacted at 25°C for 1 hour to obtain a solution containing compound 50-8, which was directly used in the next reaction.

Preparation of compound 50-9

Under a nitrogen atmosphere, triethylamine (416.2 μL, 2.99 mmol) was added to the reaction mixture from the previous step (pH > 8), followed by methyl 4-aminopyridine-2-carboxylate (136.68 mg, 0.90 mmol). The reaction mixture was stirred at 25°C for 1 hour. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to obtain the crude product, which was then isolated and purified by silica gel column chromatography (petroleum ether/ethyl acetate = 5/1-1/2) to obtain compound 50-9.

Preparation of Example 50

Under a nitrogen atmosphere, compound 50-9 (74 mg, 0.14 mmol) was added to a 7.0 M ammonia methanol solution (3 mL, 21 mmol), and the reaction mixture was incubated at 25°C for 12 hours. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to obtain the crude product, which was then purified via reverse phase chromatography (column: Boston Green ODS 150*30mm*5um; mobile phase: [A: _H2O (0.225% FA); B: ACN]; B%: 48.00%-78.00%, 11.00 min; flow rate: 25.00 mL/min) to obtain Example 50.

¹ H NMR (400MHz, DMSO-d ₆ ): δ10.62(s,1H),8.48(d,J＝5.6Hz,1H),8.27(d,J＝2.0Hz,1H),8.05(s,1H),7.81(d d,J＝5.6,2.0Hz,1H),7.60(d,J＝2.0Hz,1H),7.51-7.37(m,1H),7.35-7.23(m,1H),5. 16(d,J＝10.8Hz,1H),4.65(d,J＝2.0Hz,2H),4.40-4.30(m,1H),4.24(dd,J＝4.4,2.4H z,2H),3.55(s,1H),3.52-3.49(m,1H),2.82(s,1H),1.64(s,3H),0.83-0.64(m,3H);

m/z(ESI):[M+H] ⁺ =512.1.

Example 51

Preparation of Example 51

Compound 1-18 (200 mg, 0.44 mmol) was dissolved in DMF (2 mL), and potassium carbonate (180.50 mg, 1.31 mmol), chloromethyl methyl sulfide (126.13 mg, 1.31 mmol), and potassium iodide (72.27 mg, 0.44 mmol) were added. The reaction was allowed to react at 25°C for 1 hour. The reaction solution was added to 20 mL of aqueous solution and extracted with dichloromethane. The organic phase was separated and washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and dried. The crude product was purified by reverse phase preparative purification (Phenomenex luna C18 150×25 mm×10 μm; flow rate: 25 mL/min; gradient: 52%-72% Bover 10 min; mobile phase A: 0.225% aqueous methanoic acid, mobile phase B: acetonitrile) to obtain Example 51.

¹ H NMR (400 MHz, DMSO-d ₆ ): δ10.71(s,1H),8.49(d,J＝6.0Hz,1H),8.27(d,J＝2.4Hz,1H),8.07(d,J＝2.0Hz ,1H),7.82(dd,J＝2.0,5.6Hz,1H),7.63(d,J＝2.0Hz,1H),7.21(d,J＝6.4Hz,2H),5 .48-5.38(m,1H),5.37-5.31(m,1H),5.12(d,J＝10.4Hz,1H),4.38(dd,J＝7.6,10. 0Hz,1H),2.83(t,J＝7.6Hz,1H),2.22(s,3H),1.62(s,3H),0.73(d,J＝6.0Hz,3H);

m/z(ESI):[M+H] ⁺ =520.1.

Example 52

Preparation of Example 52

Compound 52-1 (70 mg, 0.26 mmol), compound 1-18 (120 mg, 0.26 mmol), potassium carbonate (110 mg, 0.80 mmol) and sodium iodide (50 mg, 0.33 mmol) were dissolved in DMF (5 mL) and stirred at room temperature for 12 hours. The reaction solution was poured into water (100 mL) to quench the mixture, and then extracted with ethyl acetate (100 mL x 2). The organic phase was washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and dried. The crude product was purified by reverse transpiration (Phenomenex luna C18 150×25 mm×10 μm; flow rate: 25 mL/min; gradient: 45%-75% B over 15 min; mobile phase A: 0.225% aqueous methanoic acid, mobile phase B: acetonitrile) to obtain Example 52.

¹ H NMR (400MHz, DMSO-d ₆ ): δ10.72(s,1H),8.49(d,J＝5.6Hz,1H),8.26(d,J＝2.0Hz,1H),8.08(s,1H),7.84(dd,J＝2.0,5.6Hz,1H),7.64(s,1H),7.61-7.52(m,1H ),7.43-7.35(m,1H),5.17(d,J＝10.2Hz,1H),4.79-4.71(m,1H),4.26-4.18(m,1H),2.73-4.64(m,1H),1.60(s,3H),0.78–0.70(m,3H);

m/z (ESI): [M+H] ⁺ = 534.0.

Examples 53 and 54

Preparation of compound 53-1

A solution of ethynylmagnesium bromide in tetrahydrofuran (1.9 mL, 0.96 mmol, 0.5 M) was slowly added dropwise to a solution of 50-5 (300 mg, 0.73 mmol) in THF (10 mL) at 0°C. The reaction mixture was naturally warmed from 0°C to 25°C and stirred for 2 hours. After completion, aqueous ammonium chloride (3 mL) was slowly added dropwise to the reaction mixture at 0°C to quench the reaction. The mixture was diluted with water (10 mL) and extracted with ethyl acetate (10 mL x 3). The organic phase was dried over anhydrous sodium sulfate, filtered, and dried by rotary evaporation. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 5:1-3:1) to obtain compound 53-1.

Preparation of compound 53-2

Diethylaminosulfur trifluoride (0.1 mL, 607.73 μmol) was slowly added dropwise to a solution of 53-1 (220 mg, 506.45 μmol) in DCM (5 mL) at -20°C. The reaction mixture was naturally warmed from 0°C to 25°C and allowed to react for 2 hours. The reaction mixture was slowly added dropwise to saturated sodium bicarbonate to quench the mixture. The mixture was extracted with dichloromethane (5 mL x 3). The organic phase was dried and concentrated, and the crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 10:1 to 3:1) to obtain compound 53-2.

Preparation of compound 53-3

Compound 53-2 (195 mg, 0.45 mmol) was dissolved in DCM (5 mL), trifluoroacetic acid (1 mL) was added, and the mixture was reacted at 25° C. for 1 hour. The reaction solution was directly concentrated to obtain compound 53-3.

Preparation of compound 53-4

Compound 53-3 (150 mg, 0.39 mmol) and DMF (3.1 μL, 0.04 mmol) were dissolved in DCM (5 mL), and oxalyl chloride (169.4 μL, 1.97 mmol) was slowly added under ice-cooling. The reaction mixture was stirred at room temperature for 3 hours. The reaction mixture was concentrated to obtain compound 53-4.

Preparation of compound 53-5

Compound 53-4 (140 mg, 0.35 mmol) was dissolved in DCM (0.5 mL). Triethylamine (0.2 mL, 1.76 mmol) and methyl 4-aminopyridine-2-carboxylate (53.42 mg, 0.35 mmol) were slowly added under ice-cooling. The mixture was stirred at room temperature for 3 hours. The reaction solution was concentrated, and the crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 1:1) to obtain compound 53-5.

Preparation of Examples 53 and 54

Compound 53-5 (20 mg, 0.04 mmol) and ammonia methanol (16.55 mg, 0.97 mmol) were dissolved in MeOH (0.5 mL), and the reaction was stirred at 25°C for 3 h. The reaction solution was concentrated, and the crude product was purified by reverse phase preparative chromatography (Phenomenex luna C18 150×25 mm×10 μm; flow rate: 25 mL/min; gradient: 49%–69% B over 10 min; mobile phase A: 0.225% aqueous methanoic acid, mobile phase B: acetonitrile) to obtain two isomers.

Example 53 (retention time is short):

¹ H NMR (400 MHz, DMSO-d ₆ ): δ10.56(s,1H),8.48(d,J＝5.6Hz,1H),8.26(d,J＝2.0Hz,1H),8.06(d,J＝2 .4Hz,1H),7.82(dd,J＝2.0,5.2Hz,1H),7.67-7.56(m,2H),7.40-7.35(m,1H ),6.85-6.69(m,1H),5.16(d,J＝10.0Hz,1H),4.46(t,J＝9.2Hz,1H),4.28(d d,J＝2.0,5.0Hz,1H),2.94-2.85(m,1H),1.66(s,3H),0.76(d,J＝7.2Hz,3H);

m/z(ESI):[M+H] ⁺ =500.2.

Example 54 (longer retention time):

¹ H NMR: (400 MHz, DMSO-d ₆ ): δ10.58(s,1H),8.48(d,J＝5.2Hz,1H),8.27(d,J＝2.0Hz,1H),8.06(d,J＝1.2 Hz,1H),7.86(dd,J＝2.2,5.6Hz,1H),7.65-7.56(m,2H),7.40(dd,J＝4.2,8.8H z,1H),6.93-6.77(m,1H),5.23(d,J＝10.0Hz,1H),4.63(t,J＝9.2Hz,1H),4.09 (dd,J＝2.2,5.0Hz,1H),2.67-2.61(m,1H),1.67(s,3H),0.69(d,J＝6.4Hz,3H);

m/z (ESI): [M+H] ⁺ = 500.2.

Example 55

Preparation of compound 52-1

Under a nitrogen atmosphere, a solution of n-butyllithium in n-hexane (33 mL, 82.50 mmol) was slowly added dropwise to a solution of compound 55-1 (13.3 mL, 71.28 mmol) in THF (80 mL) at -70°C. After stirring for 1 hour, compound 55-2 (23 g, 109.62 mmol) was slowly added dropwise to the reaction system. The temperature was returned to 20°C and stirring was continued for 15 hours. The reaction solution was poured into an icy aqueous ammonium chloride solution (100 mL), and the mixture was extracted with petroleum ether (200 mL x 2). The organic phases were combined, washed once with water and once with saturated brine, dried over anhydrous sodium sulfate, filtered, and spin-dried to obtain the crude product. The crude product was dissolved in 50 mL of petroleum ether, filtered through silica gel powder (10 cm thick), and the filtrate was concentrated to dryness to obtain compound 52-1.

Preparation of compound 55-3

Compound 52-1 (10 g, 37.15 mmol) was dissolved in tetrahydrofuran (10 mL) and water (40 mL). Indium powder (6 g, 52.26 mmol) was added and ultrasonically vibrated for 8 hours. Aqueous formaldehyde solution (16 g, 197.14 mmol) was added to the reaction mixture and stirring continued for 32 hours. The reaction mixture was diluted with ethyl acetate (300 mL), filtered, and the filtrate was allowed to stand for separation. The upper ethyl acetate phase was separated and washed once with saturated brine. The organic phase was dried over anhydrous sodium sulfate, filtered, and spin-dried to obtain the product, which was purified by silica gel column chromatography to obtain compound 55-3.

Preparation of compound 55-4

Compound 55-3 (500 mg, 2.27 mmol) and pyridine (0.5 mL, 6.18 mmol) were dissolved in DCM (10 mL). Trifluoromethanesulfonic anhydride (0.5 mL, 3.01 mmol) was slowly added under ice-cooling and stirred at room temperature for 1 hour. The reaction mixture was diluted with ethyl acetate (200 mL) and washed sequentially with water (100 mL × 2) and saturated brine (100 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and dried to obtain compound 55-4.

Preparation of Example 55

Compound 55-4 (130 mg, 0.37 mmol) and compound 1-18 (170 mg, 0.37 mmol) were dissolved in DMF (5 mL), potassium carbonate (180 mg, 1.30 mmol) was added, and the mixture was stirred at room temperature for 12 hours. The reaction solution was poured into water (100 mL), and the mixture was extracted with ethyl acetate (100 mL x 2). The organic phase was dried over anhydrous sodium sulfate, filtered, and dried to obtain the crude product, which was purified by reverse preparation (Phenomenex luna C18 150×25 mm×10 μm; flow rate: 25 mL/min; gradient: 42%-72% B over 15 min; mobile phase A: 0.225% aqueous methanoic acid, mobile phase B: acetonitrile) to obtain Example 55.

m/z(ESI):[M+H] ⁺ =548.1.

Examples 56 and 57

Preparation of compound 56-1

Under nitrogen atmosphere, triethylamine (1.1 mL, 7.87 mmol) was added to 24-3 (314 mg, 0.79 mmol) in 3 mL of dichloromethane, followed by the addition of a solution of compound 22-5 (198.83 mg, 1.02 mmol) in 3 mL of DCM. The mixture was stirred at 26°C for 2 hours. After completion of the reaction, 10 mL of water was added to quench the reaction, followed by extraction with dichloromethane (10 mL × 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product, which was purified by silica gel column chromatography (petroleum ether/ethyl acetate = 1/1) to obtain compound 56-1.

Preparation of Examples 56 and 57

Under nitrogen atmosphere, trifluoroacetic acid (800.8 μL, 10.78 mmol) was added to a 9 mL dichloromethane solution of compound 56-1 (300 mg, 0.54 mmol), and the reaction solution was reacted at 25 degrees for 2 hours. After completion of the reaction, the reaction solution was concentrated to obtain a crude product, which was purified by reverse phase preparative chromatography (Waters Xbridge 150×25mm _× 5um, water (water( _NH4HCO3 )-ACN, 38%-68% over 12 min) to obtain a mixture of Example 56 and Example 57. SFC separation (column: DAICEL CHIRALCEL OX (250mm*30mm, 10um); mobile phase: [A: _CO2 ; B: EtOH (0.1% _{NH3H2O )} ]; B%: 15.00%-15.00%, 60.00min; flow rate: 150.00g/min) gave two single isomers.

Example 56 (retention time = 1.497 minutes)

¹ H NMR (400MHz, DMSO-d ₆ ): δ10.36(s,1H),8.71-8.56(m,1H),8.09-7.88(m,1H),7.50-7.37(m ,1H),7.29-7.08(m,2H),5.44-5.29(m,1H),5.16-5.04(m,1H),4.70-4 .60(m,1H),4.57-4.48(m,1H),4.40-4.26(m,1H),3.70-3.57(m,2H), 3.49-3.39(m,1H),2.91-2.75(m,1H),1.61(s,3H),0.81-0.63(m,3H);

m/z (ESI): [M+H] ⁺ = 517.3.

Example 57 (retention time = 1.655 minutes)

¹ H NMR (400MHz, DMSO-d ₆ ): δ10.37(s,1H),8.72-8.59(m,1H),7.97(dd,J＝2.4,8.6Hz,1H),7.42(d,J＝8.5H z,1H),7.29-7.10(m,2H),5.34(d,J＝4.8Hz,1H),5.10(d,J＝10.5Hz,1H),4.64(br t,J＝5.8Hz,1H),4.59-4.47(m,1H),4.31(dd,J＝7.5,10.3Hz,1H),3.71-3.57(m,2H),3 .44(td,J＝5.9,11.4Hz,1H),2.83(t,J＝7.3Hz,1H),1.61(s,3H),0.74(d,J＝6.0Hz,3H);

m/z: [M+H] ⁺ = 517.1.

Examples 58 and 59

Preparation of compound 58-1

To a solution of compound 50-1 (500 mg, 1.47 mmol) in DCM (5 mL) were added DMF (10 mg, 0.15 mmol) and oxalyl chloride (0.4 mL, 4.41 mmol). The mixture was stirred at 25°C for 1 hour. The mixture was concentrated, and under a nitrogen atmosphere, triethylamine (0.6 mL, 4.35 mmol) and the above concentrate were added sequentially to a solution of compound 56-5 (337.89 mg, 1.74 mmol) in 10 mL of dichloromethane. The reaction mixture was stirred at 26°C for 16 hours. After completion of the reaction, the mixture was diluted with 30 mL of water and extracted with ethyl acetate (30 mL x 2). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product, which was then purified by silica gel column chromatography (petroleum ether/ethyl acetate = 10/1 to 1/1) to obtain compound 58-1.

¹ H NMR (400 MHz, CDCl ₃ ): δ＝8.56(d,J＝2.4Hz,1H),8.41(s,1H),8.13(dd,J＝2.4,8.4Hz,1H),7.50(d ,J＝8.4Hz,1H),7.09(t,J＝6.8Hz,1H),6.83-6.72(m,1H),6.31(s,1H),5.18(t ,J＝6.8Hz,1H),5.14-5.03(m,1H),4.44(t,J＝7.6Hz,1H),3.91(t,J＝7.6Hz,1H ),2.92-2.79(m,1H),1.70(s,3H),1.51(d,J＝14.4Hz,6H),0.87-0.78(m,3H);

m/z (ESI): [M+H] ⁺ = 517.2.

Preparation of compound 58-2

Under a nitrogen atmosphere, trifluoromethanesulfonic anhydride (83.5 μL, 0.50 mmol) was added to a solution of compound 58-1 (280 mg, 0.61 mmol) in 4 mL of dichloromethane at 0°C. The reaction mixture was stirred at 0°C for 2 hours. After completion, the reaction was quenched by the addition of 20 mL of water and extracted with dichloromethane (30 mL x 2). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product, which was then purified by silica gel column chromatography (petroleum ether/ethyl acetate = 10/1 to 2/1) to obtain compound 58-2.

Preparation of compound 58-3

Under nitrogen atmosphere, ethynyl[tri(propan-2-yl)]silane (0.2 mL, 0.77 mmol), cuprous iodide (3.0 mg, 0.02 mmol), triethylamine (23.4 mg, 0.23 mmol) and 1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (8.46 mg, 0.01 mmol) were added sequentially to a solution of compound 58-2 (50 mg, 0.08 mmol) in 1.5 mL of N,N-dimethylformamide. The reaction mixture was heated to 100 degrees and stirred for 16 hours. After the reaction was completed, the mixture was cooled to room temperature, diluted with 20 mL of water and 20 mL of ethyl acetate, filtered, and the filtrate was extracted with ethyl acetate (10 mL × 2). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product, which was purified by thin-layer silica gel preparation (petroleum ether/ethyl acetate = 3/1) to obtain compound 58-3.

Preparation of compound 58-4

Under a nitrogen atmosphere, trifluoroacetic acid (262 μL, 3.53 mmol) was added to a solution of compound 58-3 (120 mg, 0.18 mmol) in 2 mL of dichloromethane. The reaction was stirred at 25°C for 0.5 h. After completion of the reaction, saturated sodium bicarbonate was slowly added dropwise to adjust the pH to 7. The aqueous phase was extracted with dichloromethane (20 mL x 2). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to yield compound 58-4.

Preparation of Examples 58 and 59

Under a nitrogen atmosphere, cesium fluoride (284 mg, 1.87 mmol) was added to a solution of compound 58-4 (120 mg, 0.19 mmol) in 2 mL of N,N-dimethylformamide. The reaction was stirred at 25°C for 0.5 h. After completion of the reaction, the reaction solution was filtered, and the filtrate was purified by reverse phase preparative chromatography (0.1% FA/ACN, 55%-65% over 5 min) to afford a mixture of Examples 58 and 59. SFC separation (column: DAICEL CHIRALCEL OX (250 mm x 30 mm, 10 μm); mobile phase: [A: CO ₂ ; B: EtOH (0.1% NH ₃ H ₂ O)]; B%: 20.00%-20.00%, 4.00 min; flow rate: 150.00 g/min) afforded two single isomers:

Example 58 (retention time = 1.587 minutes)

¹ H NMR (400MHz, DMSO-d ₆ ): δ10.39(s,1H),8.64(d,J＝2.4Hz,1H),7.99(dd,J＝2.4,8.6Hz,1H),7.61-7.50(m,1H),7.4 2(d,J＝8.6Hz,1H),7.30(dd,J＝4.4,8.4Hz,1H),5.34(d,J＝4.8Hz,1H),5.18(d,J＝10.0Hz,1H) ,5.03(s,1H),4.64(t,J＝6.0Hz,1H),4.53(td,J＝4.0,6.8Hz,1H),4.38(dd,J＝7.8,10.0Hz,1 H),3.68-3.58(m,1H),3.43(td,J＝6.0,11.6Hz,1H),2.97-2.84(m,1H),1.60(s,3H),0.73(br d,J=6.0Hz,3H);

m/z (ESI): [M+H] ⁺ = 485.2.

Example 59 (retention time = 1.726 minutes)

¹ H NMR (400MHz, DMSO-d ₆ ): δ10.45(s,1H),8.70-8.65(m,1H),8.02-7.95(m,1H),7.61-7.49(m,1H),7.42(d, J＝8.4Hz,1H),7.32(dd,J＝4.4,8.4Hz,1H),5.35(d,J＝4.8Hz,1H),5.20(d,J＝10.0Hz ,1H),5.02(s,1H),4.65(t,J＝6.0Hz,1H),4.58-4.49(m,1H),4.39(dd,J＝7.8,9.8Hz ,1H),3.68-3.59(m,1H),3.48-3.41(m,1H),2.96-2.82(m,1H),1.60(s,3H),0.73(br d,J=6.0Hz,3H);

m/z: [M+H] ⁺ = 485.1.

Example 60

Preparation of compound 60-2

Under a nitrogen atmosphere, a solution of compound 60-1 (0.64 g, 5.60 mmol) in acetonitrile was added dropwise to a solution of cuprous iodide (0.1 g, 0.51 mmol) and ethynyltrimethylsilane (0.7 mL, 5.09 mmol) in acetonitrile (5 mL). The reaction mixture was allowed to react at 25°C for 16 hours. After completion of the reaction, the reaction mixture was filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product, which was purified by silica gel column chromatography (petroleum ether/ethyl acetate = 10/1) to afford compound 60-2.

Preparation of compound 60-3

Under nitrogen atmosphere, deuterated lithium aluminum hydride (0.13 g, 3.04 mmol) was slowly added portionwise to a solution of compound 60-2 (0.7 g, 3.80 mmol) in 20 mL of THF at 0°C. The reaction mixture was allowed to react at zero°C for 1 hour. After completion, the reaction was quenched by slowly adding 1.5 mL of deuterated water dropwise at zero°C. The mixture was stirred for 10 minutes, then heated to 26°C, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product, which was purified by silica gel column chromatography (petroleum ether/ethyl acetate = 10/1) to afford compound 60-3.

Preparation of compound 60-4

Under a nitrogen atmosphere, cyanomethylene tributylphosphine (210.17 mg, 0.87 mmol) was added to a 2 mL dioxane solution containing compound 1-18 (80 mg, 0.17 mmol) and compound 60-3 (170 mg, 1.39 mmol). The reaction mixture was heated to 40°C and stirred for 16 hours. After completion of the reaction, the reaction mixture was cooled to room temperature, diluted with 10 mL of water, and extracted three times with ethyl acetate (5 mL x 3). The combined organic phases were washed with 10 mL of saturated sodium chloride solution, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to obtain the crude product, which was then purified by silica gel column chromatography (petroleum ether/ethyl acetate = 10/1 to 1/1) to obtain compound 60-4.

Preparation of Example 60

Under nitrogen atmosphere, tetrabutylammonium fluoride tetrahydrofuran solution (0.17 mL, 0.17 mmol) was added to a 1 mL THF solution of compound 60-4 (50 mg, 0.09 mmol), and the reaction solution was reacted at 25 degrees for 1 hour. After completion of the reaction, 10 mL of water was added to quench the reaction, and the mixture was extracted with ethyl acetate (5 mL × 3). The combined organic phases were dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated to obtain the crude product, which was purified by reverse phase preparation ((Phenomenex luna C18 150×25 mm×10 um); flow rate: 25 mL/min; gradient: 42%-72% B over 10 min; mobile phase A: 0.225% aqueous methanoic acid, mobile phase B: acetonitrile) to obtain Example 60.

¹ H NMR (400MHz, DMSO-d ₆ ): δ10.81-10.60(m,1H),8.55-8.46(m,1H),8.33-8.26(m,1H),8.11-8.01(m,1H),7.90-7.80(m,1H),7.69-7.58(m,1H),7.27- 7.07(m,2H),5.19-5.05(m,1H),4.44-4.26(m,1H),2.92-2.79(m,2H),2.70-2.60(m,2H),1.72-1.57(m,3H),0.79-0.64(m,3H);

m/z (ESI): [M+H] ⁺ = 514.0.

Example 61

Preparation of compound 61-1

To a solution of compound 50-5 (500 mg, 1.22 mmol) in tetrahydrofuran (5 mL) were added elemental iodine (396.70 mg, 2.45 mmol) and aqueous ammonia (2.5 mL, 30% content), and the reaction mixture was stirred at 30°C for 10 hours. The reaction mixture was quenched with 30 mL of water and extracted with ethyl acetate (30 mL × 2). The combined organic phases were washed sequentially with saturated aqueous sodium sulfite (30 mL × 2) and saturated aqueous sodium chloride (30 mL × 1). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by silica gel chromatography (petroleum ether:ethyl acetate = 10:1) to obtain compound 61-1.

¹ H NMR (400MHz, CDCl ₃ ): δ = 7.45 (q, J = 9.0Hz, 1H), 7.24 (dd, J = 4.3, 8.8Hz, 1H), 4.82 (d, J = 9.2Hz, 1H), 4. 15-4.12(m,1H),2.94-2.89(m,1H),1.66(s,3H),1.39(s,9H),0.88-0.79(m,3H).

Preparation of compound 61-2

To a solution of compound 61-1 (300 mg, 0.74 mmol) in DCM (10 mL) was added trifluoroacetic acid (2.00 mL), and the mixture was stirred at 20° C. for 5 hours. After the reaction was completed, the reaction solution was directly concentrated under reduced pressure to obtain compound 61-2.

m/z(ESI):[MH] ^- =348.0.

Preparation of compound 61-3

Compound 61-2 (260 mg, 0.74 mmol) and methyl 4-aminopyridine-2-carboxylate (169.90 mg, 1.12 mmol) were dissolved in acetonitrile (5 mL), and N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (313.31 mg, 1.12 mmol) was added. The mixture was reacted at 20°C for 16 hours. 30 mL of water was added to the reaction solution, and the mixture was extracted with ethyl acetate (30 mL x 3). The combined organic phases were washed with saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by silica gel chromatography (petroleum ether/ethyl acetate = 3/7) to obtain compound 61-3.

m/z (ESI): [M+H] ⁺ = 484.2.

Preparation of Example 61

Compound 61-3 (100 mg, 0.21 mmol) was added to ammonia methanol solution (2 mL, 6 M) and reacted at 20°C for 16 hours. After completion of the reaction, the reaction solution was directly concentrated under reduced pressure, and the crude product was purified via reverse phase chromatography (Phenomenex luna C18 150×25 mm×10 μm; flow rate: 25 mL/min; gradient: 42%-62% B over 10 min; mobile phase A: 0.225% aqueous methanoic acid, mobile phase B: acetonitrile) to obtain Example 61.

¹ H NMR (400MHz, DMSO-d ₆ ): δ10.68(s,1H),8.50(d,J＝5.6Hz,1H),8.30(d,J＝2.0Hz,1H),8.08(d,J＝2.0Hz,1H),7.95-7.87(m,1H),7.85(dd,J＝2.0,5.6Hz,1H),7.62(br s,1H),7.49(dd,J＝4.4,8.8Hz,1H),5.26(d,J＝9.2Hz,1H),4.31(t,J＝8.8Hz,1H),2.91(t,J＝7.6Hz,1H),1.62(s,3H),0.79(d,J＝6.4Hz,3H);

m/z (ESI): [M+H] ⁺ = 469.0.

Experimental Example 1 Manual patch clamp method to detect the inhibitory activity of the compound of the present invention on voltage-gated sodium channel NaV1.8

Cell culture and passaging: CHO cells stably expressing human NaV1.8 were cultured in Ham's F-12 medium containing 10% fetal bovine serum, 10 μg/mL blasticidin, 200 μg/mL hygromycin B, and 100 μg/mL zeocin. The cell culture temperature was 37°C and the carbon dioxide concentration was 5%. During cell passaging, the old culture medium was removed and the cells were washed once with PBS. Then, 0.25%-Trypsin-EDTA solution was added and incubated at 37°C. When the cells were observed to fall off from the bottom of the dish, an appropriate amount of complete culture medium preheated at 37°C was added. After the cells were blown off from the bottom of the dish, they were transferred to a sterile centrifuge tube and centrifuged at 1000 rpm for 5 minutes to collect the cells. The cells were then seeded into 6 cm cell culture dishes (2.5×10 ⁵ cells/dish, 5 mL culture medium) for expansion or maintenance culture. To maintain the electrophysiological activity of the cells, the cell density must not be lower than 80%.

Before patch clamp analysis, 0.25% trypsin-EDTA solution was added to CHO cells stably expressing human NaV1.8 to detach the cells and count them. 6.5×10 ³ cells were attached to coverslips and cultured in 24-well plates (final volume 500 μL). The cells were tested 18 hours later.

Compound sample preparation: Test compounds were prepared in dimethyl sulfoxide (DMSO) to a 100 mM stock solution. The solution was then diluted to various working concentrations in an extracellular solution containing 100 nM tetrodotoxin (TTX) (140 mM NaCl, 3.5 mM KCl _{, 1 mM} MgCl₂· _6H₂O , ₂ mM CaCl₂·2H₂O, ₁₀ mM D-glucose, 10 mM HEPES, _{1.25 mM} NaH₂PO₄·2H₂O, pH adjusted to 7.4 with NaOH). The DMSO concentration in each working solution was 0.1%. The working solutions were sonicated for 20 min prior to testing.

Patch clamp testing: First, a recording electrode is drawn from a glass capillary using a microelectrode puller. The electrode, filled with intracellular solution (50mM CsCl, 10mM NaCl, 10mM HEPES, 60mM CsF, 20mM EGTA, adjusted to pH 7.2 with CsOH), is then loaded into a microelectrode holder. Under an inverted microscope, the microelectrode manipulator is used to bring the recording electrode into contact with the cell. Negative pressure is applied to create a GΩ seal. Fast capacitance compensation is then applied, followed by continued negative pressure to rupture the cell membrane and establish whole-cell recording mode. Finally, slow capacitance compensation is performed and the relevant parameters are recorded. No leakage compensation is performed.

When the sodium current recorded by the whole cell is stable, the drug is administered, and each drug concentration is acted on for 5 minutes (or the current is stable) before the next concentration is detected. The coverslip with cells is placed in a recording bath under an inverted microscope, and the blank control external solution and the test compound working solution are flowed through the recording bath from low concentration to high concentration in sequence by gravity perfusion to act on the cells. A peristaltic pump is used for liquid exchange during recording. The current detected in the external solution without compound for each cell serves as its own control group. Each concentration is independently repeated 2 times. All electrophysiological tests are carried out at room temperature. Specifically, 2 concentrations (preliminary screening) or 5 concentrations (IC ₅₀ value calculation) are set for each test compound. The inhibitory activity of the test compound on NaV1.8 sodium channels is determined by calculating the relative percentage of the peak current generated after the test compound treats the cells and the peak current generated by the control group cells.

Voltage stimulation protocol for whole-cell patch-clamp recording of NaV1.8 sodium currents: After whole-cell patch-clamp formation, the cell is voltage-clamped at -120 mV. The voltage is first stepped from -130 mV to -10 mV in 10 mV steps and held for 5 seconds. A depolarizing pulse of 0 mV is then applied to obtain the half-inactivation voltage (Vhalf). The resting and half-inactivated states of sodium current are monitored using a double-pulse protocol. A first depolarizing pulse (TP1) is applied to 0 mV for 50 ms to measure the resting sodium current. The voltage is then adjusted to Vhalf and held for 5 seconds. The voltage is then returned to -120 mV and held for 20 ms. A second depolarizing pulse (TP2) is applied to 0 mV for 50 ms to measure the half-inactivated sodium current. Finally, the voltage is returned to the clamping voltage of -120 mV. Data are collected repeatedly every 20 ms to observe the effects of drugs on the peak sodium currents in the two distinct states.

The inhibitory activity of the example compounds on Na _V 1.8 channels was determined by the above test. The inhibition rate and IC ₅₀ at a concentration of 1 nM are shown in Tables 1 and 2.

Table 1. Inhibitory activity of the compounds in the examples on NaV1.8 channels (inhibition rate)

Table 2. Inhibitory activity of the example compounds on NaV1.8 channels (IC ₅₀ )

The experimental results show that the compounds of the present invention have a good inhibitory effect on NaV1.8.

Experimental Example 2: In vivo pharmacokinetic study was conducted on rats by single intravenous injection or oral administration.

Experimental method: Six male SD rats weighing 200-300 g were divided into two groups. One group received a single oral dose of 5 mg/kg, and the other group received a single tail vein dose of 1 mg/kg. The animals that received oral administration fasted overnight and resumed eating 4 hours after administration. The animals that received intravenous administration were free to eat. Blood was collected at 0.083, 0.25, 0.5, 1, 2, 4, 6, 8 and 24 hours after administration. After pretreatment, the plasma samples were detected by LC/MS/MS in MRM mode, and a suitable standard curve was established to quantify the target compound in the plasma sample to obtain the drug concentration-time curve. The pharmacokinetic parameters were calculated using the non-atrioventricular membrane model of WinNonlin software. The experimental results are shown in the following table:

Table 3. Pharmacokinetic parameters of the compounds of the present invention in rats

The experimental results show that the compounds of the present invention have high blood concentrations, high exposure amounts, and low clearance rates in rats, and have obvious pharmacokinetic advantages.

Experimental Example 3: In vivo pharmacokinetic study was conducted on mice by single intravenous injection or oral administration.

Experimental Methods: Six male C57BL/6 mice weighing 20-30 g were divided into two groups. One group received a single dose of 5 mg/kg via the tail vein, and the other group received a single oral dose of 1 mg/kg. Both groups of animals had free access to food. Blood was collected at 0.083, 0.25, 0.5, 1, 2, 4, 6, 8, and 24 hours after administration. After pretreatment, the plasma samples were detected by LC/MS/MS in MRM mode, and a suitable standard curve was established to quantify the target compound in the plasma sample to obtain the drug concentration-time curve. The pharmacokinetic parameters were calculated using the non-compartmental model of WinNonlin software. The experimental results are shown in the following table:

Table 4. Pharmacokinetic parameters of the compounds of the present invention in mice

The experimental results show that the compounds of the present invention have high blood concentrations and high exposure in mice, and have pharmacokinetic advantages.

Experimental Example 4: In vivo pharmacokinetic study was conducted on dogs by single intravenous injection or oral administration.

Experimental method: Six male beagle dogs weighing 9-11 kg were divided into two groups. One group received a single dose of 0.5 mg/kg via the tail vein, and the other group received a single oral dose of 1 mg/kg. The animals in the intravenous administration group were free to eat, while the animals in the oral administration group fasted overnight and resumed eating 4 hours after administration. Blood was collected from the intravenous administration group at 0.083, 0.25, 0.5, 1, 2, 4, 8, 24 and 48 hours after administration, and from the oral administration group at 0.25, 0.5, 1, 2, 4, 8, 24, 32 and 48 hours after administration. After pretreatment, the plasma samples were detected by LC/MS/MS in MRM mode, and a suitable standard curve was established to quantify the target compound in the plasma sample to obtain the drug concentration-time curve. The pharmacokinetic parameters were calculated using the non-compartmental model of WinNonlin software. The experimental results are shown in the following table:

Table 5. Pharmacokinetic parameters of the compounds of the present invention in dogs

The experimental results show that the example compounds of the present invention have low clearance rate in dogs, high exposure, high oral bioavailability, and have pharmacokinetic advantages.

Experimental Example 5: In vivo pharmacokinetic study was conducted on monkeys by single intravenous injection or oral administration.

Six male crab-eating macaques weighing 3.0-3.3 kg were divided into two groups. One group received a single intravenous dose of 1 mg/kg, and the other group received a single oral dose of 2 mg/kg. The animals in the intravenous administration group were free to eat, while the animals in the oral administration group fasted overnight and resumed eating 4 hours after administration. Blood was collected from the intravenous administration group at 0.083, 0.25, 0.5, 1, 2, 4, 8, 24 and 48 hours after administration, and from the oral administration group at 0.25, 0.5, 1, 2, 4, 8, 24, 32 and 48 hours after administration. After pretreatment, the plasma samples were detected by LC/MS/MS in MRM mode, and a suitable standard curve was established to quantify the target compound in the plasma sample to obtain the drug concentration-time curve. The pharmacokinetic parameters were calculated using the non-compartmental model of WinNonlin software. The experimental results are shown in the following table:

Table 6. Pharmacokinetic parameters of the compounds of the present invention in monkeys

The experimental results show that the compounds of the present invention have low clearance rate in dogs, high exposure, high oral bioavailability, and pharmacokinetic advantages.

Experimental Example 6: Solubility of the compounds of the present invention.

Experimental method: Take 15μL of 10mM DMSO stock solution and add it to a 96-well plate (n=2) containing 485μL phosphate buffer (PBS pH 7.4), simulated fasting intestinal fluid (FaSSIF pH 6.5), simulated fed intestinal fluid (FeSSIF pH 5.0), and simulated fasting gastric fluid (FaSSGF pH 1.6), respectively, to a final concentration of 300μM (containing 3% DMSO). After adding a stirring bar to each well, seal it and shake it in a constant temperature shaker at 25℃ 1100rpm for 2h. After the shaking is completed, filter the sample in the filter plate using a vacuum filter. Take 5μL of the filtrate, add 5μL DMSO, and then add an appropriate amount of ultrapure water and acetonitrile mixture (1:1) containing internal standard to dilute the corresponding multiples. Mix well and prepare for LC-MS/MS injection analysis. Take 5 μL of 300 μM DMSO standard solution, add 5 μL of different buffers, then add an appropriate amount of ultrapure water and acetonitrile mixed solution (1:1) containing the internal standard to dilute the solution to the corresponding multiple, mix well, and perform LC-MS/MS analysis together with the sample. The experimental results are shown in the table below.

The solubility values of the test samples and standard controls are calculated according to the following formula:

Table 7. Solubility of the compounds of the present invention

The experimental results show that the example compounds of the present invention have good solubility in FaSSIF, FESSIF and FaSSGF solutions.

Experimental Example 7: CYP inhibition by the compounds of the present invention.

Experimental Method: CYP450 isoenzyme-specific probe substrates were incubated with human liver microsomes and various concentrations of the compound. Reduced nicotinamide adenine dinucleotide phosphate (NADPH) was added to initiate the reaction. After the reaction, the samples were processed and the concentrations of metabolites produced by the probe substrates were quantitatively measured using liquid chromatography-tandem mass spectrometry (LC-MS/MS). The _IC50 values were then calculated. The experimental results are shown in the following table:

Table 8 Effects of the compounds of the present invention on the activity of human liver microsomal cytochrome P450

The experimental results show that the compounds of the present invention have weak inhibition on cytochrome P450 isoenzymes and low risk of drug-drug interaction.

Experimental Example 8: Metabolic stability of the compounds of the present invention in liver microsomes.

Experimental Method: An appropriate volume of human, monkey, dog, rat, or mouse liver microsomes (final protein concentration 0.5 mg/mL) and reduced nicotinamide adenine dinucleotide phosphate (NADPH, final concentration 1 mM) were mixed in 100 mM phosphate buffer (pH 7.4). After preincubation at 37°C with shaking for 10 minutes, an appropriate volume of the target compound (final concentration 1 μM) was added to initiate the reaction. The organic solvent content did not exceed 1%. Equal volumes of the incubation solution were removed after 0.5, 5, 15, 30, and 60 minutes, and the reaction was terminated by the addition of glacial acetonitrile containing an internal standard. After centrifugation at 3220 g for 40 minutes, the supernatant was diluted with ultrapure water and quantified by LC/MS/MS. The peak area ratio of the target compound to the internal standard at 0.5 minutes was defined as 100%, and the relative percentage of the target compound remaining at the remaining time points was calculated. The natural logarithm of the remaining percentage of the target compound at each time point was linearly regressed against the incubation time to determine the slope (k). The half-life was calculated using the formula t _1/2 = -0.693/k. The experimental results are shown in the following table:

Table 9. Metabolic stability of compounds in liver microsomes according to the present invention

The experimental results show that the compounds of the present invention are relatively stable in various types of liver microsomes.

Experimental Example 9: Metabolic stability of the compounds of the present invention in hepatocytes.

Experimental method: Human, monkey, dog, rat or mouse hepatocytes (cell density 0.5×10 ⁶ cells/mL) and the target compound (final concentration 1 μM) were mixed and incubated in a 37°C incubator with shaking for 120 minutes. The organic solvent did not exceed 1%. The same volume of incubation solution was taken after 0.5, 15, 30, 60, 90 and 120 minutes of reaction, and the reaction was terminated by adding an appropriate amount of glacial acetonitrile solution containing an internal standard. After the sample was centrifuged at 3220g for 45 minutes, the supernatant was diluted with ultrapure water and quantitatively analyzed by LC/MS/MS. The peak area ratio of the target compound to the internal standard at 0.5 minutes was taken as 100%, and the relative residual percentage of the target compound at the remaining time points was calculated. The natural logarithm of the residual percentage of the target compound at each time point was linearly regressed against the incubation time to obtain the slope (k), and the half-life was calculated using the formula t _1/2 = -0.693/k. The experimental results are shown in the following table:

Table 10. Metabolic stability of compounds in the examples of the present invention in hepatocytes

The experimental results show that the compounds of the present invention are relatively stable in various types of liver cells.

Experimental Example 10: Plasma protein binding rate of the compound of the present invention.

Experimental Method: Take a 10mM stock solution in DMSO and dilute it to a 1mM working solution. Add 3μL of the working solution of the test substance to 597μL of pre-incubated plasma and mix thoroughly. The final concentration of the test substance in plasma is 5μM. The final organic solvent content of the incubation system is 0.5%. After mixing, immediately transfer 50μL of the incubation system to a new 96-well plate as the zero-point sample and treat it in the same manner as the pre-incubated samples.

After assembling the equilibrium dialysis plate and dialysis membrane, add 120 μL of the test plasma sample and pH 7.4 phosphate buffer to the air space on both sides of the dialysis membrane, seal it with a membrane, and incubate it on a vortex shaker at 300 rpm in an incubator at 37°C and 5% CO2 for 6 hours. After the incubation is completed, remove the membrane and transfer 50 μL of sample from each chamber to a new 96-well plate. The remaining incubation system is also incubated for the corresponding time under the same conditions for the stability test of the test substance. After the incubation is completed, transfer 50 μL of the incubation system for processing, and the processing method is the same as that of the dialysis incubation plasma sample.

Add 50 μL of blank plasma to the removed buffer sample. Also add an equal volume of blank buffer to the removed plasma sample, vortex for 2 minutes to mix, and add 400 μL of quencher (acetonitrile containing 0.5 μM tolbutamide) to precipitate the protein. Vortex all samples for 10 minutes, then centrifuge at 3000g, 4°C for 30 minutes. Transfer 100 μL of supernatant to a new 96-well plate and centrifuge again at 3000g, 4°C for 30 minutes. Transfer 150 μL of supernatant to a new 96-well plate, add an equal volume of pure water to mix, and use for LC-MS/MS analysis.

All data were calculated using Microsoft Excel. Peak areas were determined from extracted ion chromatograms. The free and bound percentages were calculated from the peak area ratio of the test substance to the internal standard using the following formula:

The binding percentage (%) = 100 - the free percentage. The experimental results are shown in the following table:

Table 11. Plasma protein binding rates of the compounds of the present invention

Experimental Example 11: Efficacy Test of Examples 3, 12 and 24

1. Experimental Purpose

Male SD rats and male C57 BL/6J mice were used to establish a plantar incision pain model, and the analgesic effect of the disclosed compounds was evaluated by measuring the changes in the mechanical pain threshold of the animals.

2. Experimental drugs

Example 3, Example 12 and Example 24. 5% DMSO, 10% solutol, and 85% saline were added in sequence and vortexed to mix until clear.

3. Experimental Materials and Methods

3.1 Species, strain, age, and sex of experimental animals

SD rats, 6-8 weeks old, male; C57 BL/6J mice, 6-8 weeks old, male.

3.2 Experimental Animal Grouping

After adaptive feeding, SD rats were divided into the following groups:

After adaptive feeding, C57 BL/6J mice were divided into the following groups:

3.3 Experimental methods

Plantar incision pain modeling: The experimental animals were anesthetized and fixed on the operating table in a prone position. The soles of the hind limbs were flattened upwards, fixed with surgical tape, and disinfected. A longitudinal incision was made at the heel of the sole of the foot with a sterilized blade to cut the skin fascia toward the toe end. Insert one end of the curved forceps under the lateral edge of the flexor digitorum brevis and push the forceps into the inner side of the muscle to lift the flexor digitorum brevis. Make a longitudinal incision in the muscle with a blade, making sure to cut the muscle belly in half and remove the skin. Use 7-0 sutures for suturing and disinfection. The animals were returned to their original place and recovered from the surgery overnight. Oral gavage was performed, and mechanical pain measurement was performed 3 hours after the animals were administered.

Mechanical pain measurement (Ascending method): Place the experimental animal on a mechanical pain metal grid and let it rest for 30-60 minutes. The test begins when the animal stops looking around, exploring, and becomes relatively quiet. Once the experimental animal is quiet, slowly and gently stimulate the sole of the hind limb to be tested with a Von-Frey fiber, causing the fiber to bend. The animal's paw withdrawal reaction is observed for 2-3 seconds. The animal is stimulated one by one in ascending order of fiber weight. Each fiber weight is stimulated five times continuously, with at least 10 seconds between each stimulation. If fewer than three positive reactions occur, repeat the above procedure with a larger fiber. The first time three or more positive reactions occur, the fiber used is considered the pain threshold for that animal (each animal is tested twice, and the average is taken). If the experimental animal raises its paw, avoids, or licks its paw in response to the stimulation, it is marked as positive (×). Paw withdrawal reactions caused by physical activity are not counted. If none of the above manifestations occur, it is marked as negative (○).

Fiber weight: rat 0.6, 1.0, 1.4, 2.0, 4.0, 6.0, 8.0, 10.0, 15.0 (g), cut-off value is 15.0g; mouse 0.16, 0.40, 0.60, 1.00, 1.40, 2.00 (g), cut-off value is 2.00g.

3.4 Data Analysis

After the data were summarized and counted, SPSS data statistical software (version R26.0.0.0) was used to analyze the mean and standard error. Based on the SPSS statistical results, Graph Pad (version 8.0.2) software was used to draw images, and one-way ANOVA and t-test were used to test the data.

Threshold elevation percentage (%) = [( _Gt - _G0 )/ _G0 ] x 100 (%), wherein _Gt is the plantar pain threshold of animals in the drug-treated group, and _G0 is the plantar pain threshold of animals in the vehicle group.

4. Results

The analgesic efficacy of Example 3, Example 12 and Example 24 in the rat plantar incision pain model is shown in Table 12. The analgesic efficacy of Example 3, Example 12 and Example 24 in the mouse plantar incision pain model is shown in Table 13.

Table 12 Analgesic efficacy of the disclosed compounds in the rat plantar incision pain model

Table 13 Analgesic efficacy of the disclosed compounds in the plantar incision pain model in mice

5. Conclusion

The baseline pain thresholds of rats before surgery were as follows: 3 hours after surgery, the pain threshold of Example 3 at 30 mg/kg was 7.1±0.2 g, a significant increase of 97.2% (p<0.0001) compared to the pain threshold of the vehicle control group; the pain thresholds of Example 12 at 15 and 30 mg/kg were 7.0±0.3 g and 7.5±0.2 g, respectively, significantly increasing by 94.4% (p<0.0001) and 108.3% (p<0.0001) compared to the pain thresholds of the vehicle control group. The pain threshold of Example 12 at 30 mg/kg was slightly higher than that at 15 mg/kg, and the analgesic effect was close to saturation; the pain threshold of Example 24 at 30 mg/kg was 8.1±0.3 g, a significant increase of 125% (p<0.0001) compared to the pain threshold of the vehicle control group, and the analgesic effect was significantly higher than that of Example 3 at 30 mg/kg (p<0.05).

The baseline pain thresholds of mice before surgery were as follows: 3 hours after surgery, the pain threshold of Example 3 at 60 mg/kg was 0.70±0.05 g, a significant increase of 94.4% (p<0.01) compared to the pain threshold of the vehicle control group; the pain threshold of Example 12 at 60 mg/kg was 0.78±0.06 g, a significant increase of 116.7% (p<0.001) compared to the pain threshold of the vehicle control group; and the pain thresholds of Example 24 at 30 and 15 mg/kg were 0.74±0.08 g and 0.54±0.04 g, respectively, significantly increasing by 105.6% (p<0.001) and 50.0% compared to the pain threshold of the vehicle control group. The pain threshold of Example 24 at 30 mg/kg was significantly higher than that at 15 mg/kg (p<0.05), demonstrating a clear dose-dependent analgesic effect.

Experimental Example 12: Example 61 Pharmacological Efficacy Test

1. Experimental Purpose

The mechanical allodynia test was used to evaluate the efficacy of the compounds in the mouse plantar incision pain model.

2. Experimental drugs

The compound of Example 61 was added in sequence with 5% DMSO, 10% solutol, and 85% saline, and vortexed to mix until clear.

3. Experimental Materials and Methods

3.1 Experimental animal species, strain, age, and sex: C57 BL/6 mice, 6-8 weeks old, male.

3.2 Experimental Animal Grouping

After adaptive breeding, C57 BL/6 mice were divided into the following groups:

3.3 Experimental methods

Plantar incision pain modeling: Use isoflurane to anesthetize the animal, and squeeze the animal's toes to confirm that the animal is fully anesthetized before surgery. Apply eye ointment to the animal's eyes to prevent the animal's cornea from drying out. Use iodine tincture and 70% ethanol to disinfect the sole of the left foot three times, and start the surgery after the skin is dry. Starting from 2mm from the heel, make an incision about 5mm long longitudinally toward the toe. After cutting the skin, lift the flexor digitorum brevis muscle and cause longitudinal blunt injury. Suture the wound and disinfect. After the animal is fully awake (free to move), put the animal back in the cage. 10 animals did not undergo mouse incision pain surgery, as a contrast.

Mechanical allodynia test (up-down method): One day after modeling, and 1 hour, 3 hours, and 6 hours after drug administration, the left hind paw of all model mice was tested for mechanical allodynia. Mice were individually placed in a plexiglass box with a mesh bottom to ensure that the mouse's foot was accessible for testing. The mice were acclimated for 15 minutes before testing. After acclimation, the center of the sole of the left hind paw of the mouse was tested using a test fiber. The test fiber included 8 test strengths: 2.36 (0.02g), 2.44 (0.04g), 2.83 (0.07g), 3.22 (0.16g), 3.61 (0.4g), 3.84 (0.6g), 4.08 (1g), and 4.17 (1.4g). During the test, the test fiber was pressed vertically against the skin and force was applied to bend the fiber for 6-8 seconds, with 5 seconds between each test. During the test, a rapid withdrawal of the animal's foot was recorded as a pain response. If the animal withdraws its paw when the test fiber leaves its skin, it is also recorded as a pain response. If the animal moves or walks, it is not recorded as a pain response and the test should be repeated. The test is initially performed using a force of 3.22 (0.16g). If the animal reacts to pain, the next test is performed using a test fiber of a lower force. If the animal does not react to pain, the next test is performed using a test fiber of a higher force. The maximum force of the test fiber is 4.17 (1.4g). The test results are recorded in the table below. An X indicates a pain response and an O indicates no pain response.

Mechanical allodynia is expressed as the paw withdrawal threshold (PWT) in behavioral tests in mice and is calculated according to the following formula:

50% reaction threshold (g) = (10 ^{(Xf + kδ)} )/10,000

Xf = Final test fiber value used in the test

k = table value (refer to the literature Quantitative assessment of tactile allodynia in the rat paw. J Neurosci Methods. 1994 Jul;53(1):55-63.)

δ = mean difference

3.4 Data Analysis

After the data were summarized and counted, Prism (Graph pad software, Inc.) software was used to analyze the data, and one-way ANOVA was used to test the data.

4. Results

The analgesic efficacy of Example 61 in the mouse plantar incision pain model is shown in Table 14.

Table 14 Analgesic efficacy of the disclosed compounds in the plantar incision pain model in mice

Note: Data represent Mean ± SEM, *vs vehicle control, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

5. Conclusion

At different times after administration, the compound of Example 61 had significant analgesic effect in the plantar incision pain model of mice, and had a clear dose-dependency.

In the description of this specification, the reference terms "one embodiment", "some embodiments", "example", "specific example", or "some examples" mean that the specific features, structures, materials or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic representations of the above terms do not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described can be combined in any one or more embodiments or examples in a suitable manner. In addition, those skilled in the art can combine and combine different embodiments or examples described in this specification and features of different embodiments or examples without contradiction.

Claims (13)

A compound, which is a compound represented by formula (X), or a stereoisomer, geometric isomer, tautomer, nitrogen oxide, hydrate, solvate, metabolite, pharmaceutically acceptable salt or prodrug of the compound represented by formula (X),
in: Ring A is phenyl, 5-10 membered heteroaryl or 5-10 membered heterocyclyl; Each ^Re is independently H, D, F, Cl, Br, I, CN, hydroxy, nitro, -N(R ^a ) ₂ , -(C _1-6 alkylene)N(R ^a ) ₂ , -S(O)(═NH)C _1-6 alkyl, -C(O)OR ^a , -C(O)N(R ^a ) ₂ , -(C _1-6 alkylene)C(O)N(R ^a ) ₂ , -C(O)R ^a , -S(O) ₂ NR ^a , C _1-6 alkyl, C _1-6 alkoxy, C _1-6 hydroxyalkyl, C _1-6 haloalkyl, C _1-6 haloalkoxy, C _1-6 alkylamino, C _1-6 alkylthio, C _3-6 cycloalkyl or 3-6 membered heterocyclyl, wherein the C _1-6 alkylene, C _1-6 alkyl, C 1-6 alkoxy, C _1-6 hydroxyalkyl, C _1-6 haloalkyl, C _1-6 haloalkoxy, C _1-6 alkylamino, C _1-6 alkylthio, C _3-6 cycloalkyl and 3-6 _membered heterocyclyl may be optionally substituted with 1, 2 or 3 substituents selected from D, F, Cl, Br, I, CN, hydroxy, amino, nitro, oxo, hydroxy, C _1-3 alkyl, C _1-3 alkylamino, C _1-3 haloalkyl, C _1-3 hydroxyalkyl and C _1-3 alkoxy; each ^Ra is independently H, D, hydroxy, _C1-6 alkyl, _C1-6 alkoxy, _C3-6 cycloalkyl and 3-6 membered heterocyclyl, wherein the _C1-6 alkyl, _C1-6 alkoxy, _C3-6 cycloalkyl and 3-6 membered heterocyclyl may be optionally substituted with 1, 2 or 3 substituents selected from D, F, Cl, Br, I, CN, hydroxy, amino, nitro, oxo and _C1-3 alkyl; R ¹ , R ² , R ³ , R ⁴ and R ⁵ are each independently H, D, F, Cl, Br, I, CN, hydroxy, nitro, amino, C _1-6 alkyl, C _{1-6 alkoxy, C 1-6 alkylthio, C 2-6 alkenyl} , C _2-6 alkynyl, C _3-6 cycloalkyl or 3-6 membered heterocyclyl, and the C _1-6 alkyl, C _1-6 alkoxy, C _1-6 alkylthio, C _2-6 alkenyl, C _2-6 alkynyl, C _3-6 cycloalkyl and 3-6 membered heterocyclyl may be optionally substituted with 1, 2 or 3 substituents selected from D, F, Cl, Br, I, CN, nitro, amino, hydroxy, C _1-3 alkyl, C _1-3 haloalkyl, C _1-3 hydroxyalkyl and C _1-3 alkoxy; or R ³ and R ⁵ together with the carbon atom to which they are attached form a C _3-6 cycloalkyl group; R ⁶ is CN, -S(O)C _1-6 alkyl, -S(O) ₂ C _1-6 alkyl, -CH=NOC _1-6 alkyl, C _2-6 alkynyl, C _1-6 alkylthio or -L ₁ -L ₂ -R ^c , wherein the C _1-6 alkyl, C _2-6 alkynyl and C _1-6 alkylthio may be independently and optionally substituted with 1, 2 or 3 substituents selected from D, F, Cl, Br, I, oxo and hydroxy; L ₁ is a bond, O or S; _L2 is a bond, _C1-6 alkylene or -( _C1-6 alkylene) _-C1-6 alkoxy, wherein the _C1-6 alkylene and _C1-6 alkoxy may be independently optionally substituted with 1 or 3 substituents selected from D, F, Cl, Br, I, oxo and hydroxy; R ^c is -ON=, -P(O)(C _1-6 alkyl) ₂ , C _2-6 alkynyl or -O-(3-8 membered heterocyclyl), wherein the C _1-6 alkyl, C _2-6 alkynyl and 3-8 membered heterocyclyl may be independently and optionally substituted with 1, 2, 3, 4, 5 or 6 substituents selected from D, F, Cl, Br, I, hydroxy, oxo, nitro, amino, alkylamino, C _1-6 alkyl, C _1-6 alkoxy, C _1-6 alkylthio, C _3-6 cycloalkyl and 3-6 membered heterocyclyl; ^R7 and ^R8 are each independently H, D, F, Cl, Br, I, CN, hydroxyl, amino, nitro, _C1-6 alkyl, _C1-6 alkoxy, _C3-6 cycloalkyl or 3-6 membered heterocyclyl, and the _C1-6 alkyl, _C1-6 alkoxy, _C3-6 cycloalkyl and 3-6 membered heterocyclyl may be optionally substituted with 1, 2 or 3 substituents selected from D, F, Cl, Br, I, CN, hydroxyl, amino, nitro and _C1-3 alkoxy. n is 1, 2 or 3. The compound according to claim 1, wherein ring A is phenyl, pyridazine, pyrazine, pyridine or pyrimidine. The compound according to claim 1, which is a compound represented by formula (I), (II) or (III), or a stereoisomer, geometric isomer, tautomer, nitrogen oxide, hydrate, solvate, metabolite, pharmaceutically acceptable salt or prodrug of a compound represented by formula (I), (II) or (III),
in: X is CR ¹⁰ or N; Y is CR ¹³ or N; ^R9 and ^R10 are each independently H, D, F, Cl, Br, I, CN, amino, nitro, _C1-6 alkyl, _C1-6 alkoxy, _C1-6 haloalkyl or _C1-6 haloalkoxy; R ¹¹ , R ¹² and R ¹³ are each independently H, D, F, Cl, Br, I, CN, hydroxy, nitro, -N(R ^a ) ₂ , -(C _1-6 alkylene)N(R ^a ) _{2 ,} -S(O)(═NH)C _1-6 alkyl, -C(O)OR ^a , -C(O)N(R ^a ) ₂ , -(C _1-6 alkylene)C(O)N(R ^a ) ₂ , -C(O)R ^a , -S(O) ₂ NR ^a , C _1-6 alkyl, C _1-6 alkoxy, C _1-6 hydroxyalkyl, C _1-6 haloalkyl, C _1-6 haloalkoxy, C _1-6 alkylamino, C _1-6 alkylthio, C _3-6 cycloalkyl or 3-6 membered heterocyclyl, wherein the C _1-6 alkylene, C _1-6 alkyl, C 1-6 alkoxy, C _1-6 hydroxyalkyl, C _1-6 haloalkyl, C _1-6 haloalkoxy, C _1-6 alkylamino, C _1-6 alkylthio, C _3-6 cycloalkyl and 3-6 _membered heterocyclyl may be optionally substituted with 1, 2 or 3 substituents selected from D, F, Cl, Br, I, CN, hydroxy, amino, nitro, oxo, hydroxy, C _1-3 alkyl, C _1-3 alkylamino, C _1-3 haloalkyl, C _1-3 hydroxyalkyl and C _1-3 alkoxy; Each ^Ra is independently H, D, hydroxy, _C1-6 alkyl, _C1-6 alkoxy, _C3-6 cycloalkyl or 3-6 membered heterocyclyl, and the _C1-6 alkyl, _C1-6 alkoxy, _C3-6 cycloalkyl and 3-6 membered heterocyclyl may be optionally substituted with 1, 2 or 3 substituents selected from D, F, Cl, Br, I, CN, hydroxy, amino, nitro, oxo and _C1-3 alkyl. The compound according to any one of claims 1 to 3, wherein R ¹ , R ² , R ³ , R ⁴ and R ⁵ are each independently H, D, F, Cl, Br, I, hydroxyl, methyl, ethyl, CH ₂ F, CHF ₂ , CF ₃ , -CH ₂ OCH ₃ or -CH ₂ OH. The compound according to any one of claims 1 to 4, wherein R ⁹ , R ¹⁰ and R ¹¹ are each independently H, D, F, Cl, Br, I, CN, amino, nitro, methyl, ethyl, methoxy, trifluoromethyl or trifluoromethoxy; R ¹² and R ¹³ are each independently H, D, F, Cl, Br, I, CN, hydroxyl, amino, nitro, methyl, ethyl, CH ₂ F, CHF ₂ , -OCH ₃ , -OCH ₂ CH ₃ , -C(O)NH ₂ , -C(O)NHOH, -C(O)NHOCH ₃ , -CH ₂ OH, -CH(OH)CH ₂ OH, -C(O)NHCH ₃ , -CH(OH)(CH ₃ ) ₂ , -S(O)(═NH)CH ₃ , -S(O) ₂ NH ₂ , The compound according to any one of claims 1 to 5, wherein R ⁷ and R ⁸ are each independently H, D, F, Cl, Br, I, hydroxyl, methyl, ethyl, CH ₂ F, CHF ₂ or CF ₃ . The compound according to any one of claims 1 to 6, wherein R ⁶ is CN, -S(O)C _1-3 alkyl, -S(O) ₂ C _1-3 alkyl-CH=NOC _1-3 alkyl, -C _2-6 alkynyl, C _1-3 alkylthio or -L ₁ -L ₂ -R ^c , and the C _1-3 alkyl, C _1-3 alkylthio and C _2-6 alkynyl may be independently optionally substituted with 1, 2 or 3 substituents selected from D, F, Cl, Br, I, oxo and hydroxy; L ₁ is a bond, O or S; L ₂ is a bond, C _1-3 alkylene or -(C _1-3 alkylene)-C _1-3 alkoxy; R ^c is -ON=, -P(O)(CH ₃ ) ₂ , C _2-6 alkynyl or -O-(3-8 membered heterocyclyl); the C _2-6 alkynyl and 3-8 membered heterocyclyl may be independently and optionally substituted with 1, 2, 3, 4, 5 or 6 substituents selected from D, F, Cl, Br, I, hydroxy, oxo, nitro, amino, C _1-3 alkyl, C _1-3 alkoxy, C _1-3 alkylthio, C _3-6 cycloalkyl and 3-6 membered heterocyclyl. The compound according to any one of claims 1 to 7, wherein R ₆ is CN, -S(O)CH ₃ , -S(O) ₂ CH ₃ , -C≡CH, -P(O)(CH ₃ ) _2, The compound according to claim 1-8, which has a structure shown in formula (IV) or (V):
wherein R ^c is a C _2-6 alkynyl group, which may be independently and optionally substituted by 1, ₂ , 3, 4, 5 or 6 substituents selected from D, F, Cl, Br, I, hydroxyl, nitro, amino, C _1-6 alkyl, C _1-6 alkoxy, C _1-6 alkylthio, C _3-6 cycloalkyl and 3-6 membered heterocyclyl. A compound having one of the following structures or a stereoisomer, geometric isomer, tautomer, nitrogen oxide, hydrate, solvate, metabolite, pharmaceutically acceptable salt or prodrug of a compound having one of the following structures:





A pharmaceutical composition comprising the compound according to any one of claims 1 to 10; the pharmaceutical composition optionally further comprises a pharmaceutically acceptable excipient, carrier, adjuvant or any combination thereof. Use of the compound according to any one of claims 1 to 10 or the pharmaceutical composition according to claim 11 in the preparation of a medicament for treating diseases responsive to inhibition of the voltage-gated sodium channel NaV1.8. The use according to claim 12, wherein the disease is chronic pain, intestinal pain, neuropathic pain, musculoskeletal pain, acute pain, inflammatory pain, cancer pain, idiopathic pain, postoperative pain, visceral pain, multiple sclerosis, Chuck-Marie-Doucet syndrome, incontinence, pathological cough or arrhythmia.