Macrocyclic compounds as egfr kinase inhibitors

Abstract

The present application relates to macrocyclic compounds as EGFR kinase inhibitors, specifically disclosed are compounds of formula (I) or pharmaceutically acceptable salts, stereoisomers, tautomers thereof, the preparation method, pharmaceutical compositions containing the compounds, and medical uses thereof.

Description

[0001] Cross-reference to related applications

[0002] This disclosure claims the rights and priorities of Chinese Patent Application No. 202110165647.X, filed with the State Intellectual Property Office of the People's Republic of China on February 6, 2021, and Chinese Patent Application No. 202110772169.9, filed with the State Intellectual Property Office of the People's Republic of China on July 8, 2021, the entire contents of which are hereby incorporated by reference. Technical Field

[0003] This application relates to macrocyclic compounds as EGFR kinase inhibitors, specifically disclosing compounds of Formula I or pharmaceutically acceptable salts, stereoisomers, and tautomers thereof, methods for their preparation, pharmaceutical compositions containing the compounds, and their pharmaceutical uses. Background Technology

[0004] EGFR (epidermal growth factor receptor)-TKI (tyrosine kinase inhibitor), as a small molecule inhibitor, inhibits the activation of tyrosine kinase by competitively binding to EGFR through endogenous ligands, thereby blocking the EGFR signaling pathway and ultimately producing a series of biological effects such as inhibiting the proliferation and metastasis of tumor cells and promoting tumor cell apoptosis. It is one of the main targets for lung cancer treatment.

[0005] Osimertinib (AZD9291) is a third-generation EGFR-TKI targeted drug. Although it has a high response rate against resistance caused by the T790M mutation, patients can still develop resistance (Clin Cancer Res; 21(17), 2015). In 2015, Nature Medicine, 21, 560–562, 2015, first reported an analysis of resistance in 15 patients with AZD9291. Among them, the acquisition of a third mutation, namely the EGFR C797S mutation, is one of the main mechanisms leading to resistance to the drug Osimertinib, accounting for about 40%. Providing patients with safer and more effective fourth-generation EGFR C797S / T790M inhibitors is of great research significance. Summary of the Invention

[0006] On the one hand, this application relates to compounds of formula I or their pharmaceutically acceptable salts, stereoisomers, and tautomers.

[0007]

[0008] Where X is selected from O, -N(R) c - or a single key;

[0009] L is selected from single bond, C 1-8 Alkylene, -CH= or C 2-8 alkenyl;

[0010] R 1 Selected from -NR a R b Contains 1, 2 or 3 3-10 membered heterocyclic alkyl groups selected from N, O or S, or -OC 1-8 Alkyl groups, wherein the heterocyclic alkyl group is optionally substituted with one or more R groups selected from C10. 1-8 Alkyl group, C substituted with one or more deuterium groups 1-8 Alkyl groups, C substituted with one or more halogens 1-8 Alkyl group, with one C 3-8 Cycloalkyl-substituted C 1-8 Alkyl, C 3-8 Cycloalkyl, halogen, cyano, amino, or hydroxyl;

[0011] R a R b and R c Selected independently from hydrogen and C 1-8 Alkyl or C 3-6 cycloalkyl;

[0012] R 2 Selected from hydrogen, halogen, hydroxyl, cyano, amino, nitro, C 1-8 Alkyl or C 1-8 Alkoxy, where C 1-8 Alkyl or C 1-8 The alkoxy group is optionally substituted by one or more R' groups, where R' is selected from halogen, hydroxyl, cyano, amino, or nitro groups;

[0013] R 3 C selected from hydrogen, halogens, or optionally substituted with one or more halogens 1-8 alkyl;

[0014] The condition is that when the fragment Selected from R 2 Not methyl;

[0015] And the above compounds do not contain and

[0016] On the other hand, this application relates to compounds of formula I or their pharmaceutically acceptable salts, stereoisomers, and tautomers.

[0017]

[0018] Where X is selected from O, -N(R) c -, -C(O)-, or a single bond;

[0019] L is selected from a single bond or C. 1-8 Alkylene;

[0020] R 1 Selected from -NR a R b It may contain one, two, or three 3- to 10-membered heterocyclic alkyl groups selected from N, O, or S, wherein the heterocyclic alkyl group is optionally substituted with one or more R groups selected from C. 1-8 Alkyl group, C substituted with one or more deuterium groups 1-8 Alkyl groups, C substituted with one or more halogens 1-8 Alkyl group, with one C 3-8 Cycloalkyl-substituted C 1-8 Alkyl, C 3-8 Cycloalkyl, halogen, cyano, amino, or hydroxyl;

[0021] R a R b and R c Selected independently from hydrogen and C 1-8 Alkyl or C 3-6 cycloalkyl;

[0022] R 2 Selected from hydrogen, halogen, hydroxyl, cyano, amino, nitro, C 1-8 Alkyl or C 1-8 Alkoxy, where C 1-8 Alkyl or C 1-8 The alkoxy group is optionally substituted by one or more R' groups, where R' is selected from halogen, hydroxyl, cyano, amino, or nitro groups;

[0023] R 3 C selected from hydrogen, halogens, or optionally substituted with one or more halogens 1-8 alkyl;

[0024] The condition is that when the fragment Selected from R 2 It is not methyl.

[0025] In some embodiments, X is selected from O, -NH-, or -C(O)-. In some embodiments, X is selected from O, -NH-, or a single bond. In some embodiments, X is selected from O. In some embodiments, X is selected from -NH-. In some embodiments, X is selected from -C(O)-. In some embodiments, X is selected from a single bond.

[0026] In other implementations, L is selected from single bonds, C 2-6 Alkylene, -CH= or C 2-6 Alkenyl group. In other embodiments, the L is selected from single bonds, C... 2-4 Alkylene, -CH= or C 2-4 Alkenyl group.

[0027] In some other implementations, L and X are not both selected from keys.

[0028] In other implementations, the structural segment -LX- is selected from -N(R c -, O, -CH2- or -CH=, and R 1 Selected from 3-10 membered heterocyclic alkyl groups containing 1, 2 or 3 selected from N, O or S, wherein the heterocyclic alkyl group is optionally substituted with one or more R.

[0029] In some implementations, the L is selected from a single bond or C. 1-6 Alkylene. In some embodiments, L is selected from a single bond, -CH2-, -CH2CH2-, -CH2CH2CH2-, or -CH2CH2CH2CH2-. In some embodiments, L is selected from a single bond. In some embodiments, L is selected from -CH2-. In some embodiments, L is selected from -CH2CH2-. In some embodiments, L is selected from -CH2CH2CH2-. In some embodiments, L is selected from -CH2CH2CH2CH2-.

[0030] In some implementations, the R a R b and R c Selected independently from hydrogen and C 1-6 Alkyl or C 3-6 Cycloalkyl. In some embodiments, the R... a R b and R c Selected independently from hydrogen and C 1-5 Alkyl or C 3-6 cycloalkyl; in some embodiments, the R a R b and R c Selected independently from hydrogen and C 1-4 Alkyl or C 3-5 cycloalkyl; in some embodiments, the R a R b and R c Selected independently from hydrogen and C 1-3 Alkyl or C 3-4 Cycloalkyl. In some embodiments, the R... a R band R c Each is independently selected from hydrogen or C. 1-3 Alkyl group. In some embodiments, the R... a R b and R c Each of the following is independently selected from hydrogen, methyl, ethyl, or n-propyl. In some embodiments, the R... a R b and R c Each is independently selected from hydrogen, methyl, or ethyl.

[0031] In some other implementations, the R 1 Selected from -NR a R b Contains 1, 2 or 3 3-8 membered heterocyclic alkyl groups selected from N, O or S, or -OC 1-6 The alkyl group, optionally substituted with one or more R groups, is further substituted with alkyl groups. In other embodiments, the R groups are... 1 Selected from -NR a R b Contains 1, 2 or 3 3-6 membered heterocyclic alkyl groups selected from N, O or S, or -OC 1-4 The alkyl group, optionally substituted with one or more R groups, is further substituted with alkyl groups. In other embodiments, the R groups are... 1 Selected from -NR a R b Contains 1, 2 or 3 5-6 membered heterocyclic alkyl groups selected from N, O or S, or -OC 1-3 Alkyl group, wherein the heterocyclic alkyl group is optionally substituted with one or more R groups.

[0032] In some other implementations, the R 1 Selected from -NR a R b or -OC 1-6 Alkyl; in some other embodiments, the R 1 Selected from -NR a R b or -OC 1-4 Alkyl; in some other embodiments, the R 1 Selected from -NR a R b or -OC 1-3 alkyl.

[0033] In some implementations, the R 1 The heterocyclic alkyl group is selected from 1, 2 or 3 tri-, quadri-, quinary, septial, septial, septial, octial, octial or decial heterocyclic alkyl groups selected from N, O or S, wherein the heterocyclic alkyl group is optionally substituted by one or more R groups.

[0034] In some implementations, the R1 The heterocyclic alkyl group is selected from 3-9 membered heterocyclic alkyl groups containing 1, 2 or 3 selected from N, O or S, wherein the heterocyclic alkyl group is optionally substituted by one or more R.

[0035] In some implementations, the R 1 The heterocyclic alkyl group is selected from 3-8 membered heterocyclic alkyl groups containing 1, 2 or 3 selected from N, O or S, wherein the heterocyclic alkyl group is optionally substituted by one or more R.

[0036] In some implementations, the R 1 The heterocyclic alkyl group is selected from 3-7 membered heterocyclic alkyl groups containing 1, 2 or 3 selected from N, O or S, wherein the heterocyclic alkyl group is optionally substituted by one or more R.

[0037] In some implementations, the R 1 The heterocyclic alkyl group is selected from 3- to 6-membered heterocyclic alkyl groups containing 1, 2, or 3 selected from N, O, or S, wherein the heterocyclic alkyl group is optionally substituted with one or more R groups.

[0038] In some implementations, the R 1 The heterocyclic alkyl group is selected from 4-6 membered heterocyclic alkyl groups containing 1, 2 or 3 N, O or S, wherein the heterocyclic alkyl group is optionally substituted by one or more R.

[0039] In some implementations, the R 1 The heterocyclic alkyl group is selected from 5-6 membered heterocyclic alkyl groups containing 1, 2 or 3 N, O or S, wherein the heterocyclic alkyl group is optionally substituted by one or more R.

[0040] In some implementations, the R 1 The heterocyclic alkyl group is selected from 5-membered heterocyclic alkyl groups containing 1, 2 or 3 N, O or S, wherein the heterocyclic alkyl group is optionally substituted by one or more R.

[0041] In some implementations, the R 1 The heterocyclic alkyl group is selected from 6-membered heterocyclic alkyl groups containing 1, 2 or 3 N, O or S, wherein the heterocyclic alkyl group is optionally substituted with one or more R.

[0042] In some implementations, R is selected from C. 1-7 Alkyl group, C substituted with one or more deuterium groups 1-7 Alkyl groups, C substituted with one or more halogens 1-7 Alkyl group, with one C 3-7 Cycloalkyl-substituted C 1-7 Alkyl, C 3-7 Cycloalkyl, halogen, cyano, amino, or hydroxyl.

[0043] In some implementations, R is selected from C.1-6 Alkyl group, C substituted with one or more deuterium groups 1-6 Alkyl groups, C substituted with one or more halogens 1-6 Alkyl group, with one C 3-6 Cycloalkyl-substituted C 1-6 Alkyl, C 3-6 Cycloalkyl, halogen, cyano, amino, or hydroxyl.

[0044] In some implementations, R is selected from C. 1-5 Alkyl group, C substituted with one or more deuterium groups 1-5 Alkyl groups, C substituted with one or more halogens 1-5 Alkyl group, with one C 3-6 Cycloalkyl-substituted C 1-5 Alkyl, C 3-6 Cycloalkyl, halogen, cyano, amino, or hydroxyl.

[0045] In some implementations, R is selected from C. 1-4 Alkyl groups, fully deuterated C 1-4 Alkyl groups, C substituted with one or more fluorine, chlorine, bromine or iodine compounds 1-4 Alkyl group, with one C 3-6 Cycloalkyl-substituted C 1-4 Alkyl, C 3-6 Cycloalkyl, fluorine, chlorine, bromine, iodine, cyano, amino, or hydroxyl.

[0046] In some implementations, R is selected from C. 1-3 Alkyl groups, fully deuterated C 1-3 Alkyl groups, C substituted with one or more fluorine, chlorine, bromine or iodine compounds 1-3 Alkyl group, with one C 3-5 Cycloalkyl-substituted C 1-3 Alkyl, C 3-5 Cycloalkyl, fluorine, chlorine, bromine, cyano, amino, or hydroxyl.

[0047] In some embodiments, R is selected from methyl, ethyl, n-propyl, isopropyl, -CD3, -C2D5, -C3D8, -CF3, -CH2CH2F, -CH2CHF2, -CH2CF3, -C2F5, -C3F8, and C. 3-5 Cycloalkyl-substituted methyl, cyclopropyl, cyclobutyl, cyclopentyl, fluorine, chlorine, bromine, cyano, amino, or hydroxyl groups.

[0048] In some embodiments, R is selected from methyl, ethyl, -CD3, -C2D5, -CF3, -CH2CH2F, -CH2CHF2, -CH2CF3, -C2F5, cyclopropylmethyl, cyclopropyl, fluorine, chlorine, bromine, cyano, amino, or hydroxyl.

[0049] In some embodiments, R is selected from methyl, ethyl, -CD3, -CF3, cyclopropyl, cyclopropylmethyl, cyano, amino, or hydroxy.

[0050] In some other embodiments, R is selected from C. 1-3 Alkyl groups, fully deuterated C 1-3 Alkyl groups, C substituted with one or more fluorine, chlorine, bromine or iodine compounds 1-3 Alkyl group, with one C 3-5 Cycloalkyl-substituted methyl or C 3-5 Cycloalkyl. In some embodiments, R is selected from methyl, ethyl, -CD3, -CF3, cyclopropyl, or cyclopropylmethyl.

[0051] In some implementations, the R 1 The heterocyclic alkyl group is selected from azirrobutyl, tetrahydropyrrolyl, piperidinyl, azirroheptyl, imidazolinyl, pyrazolinyl, piperazinyl, hexahydropyrimidinyl, oxazolinyl, isoxazolinyl, thiazolinyl, isothiazolinyl, or morpholinyl, and the heterocyclic alkyl group is optionally substituted with one or more R.

[0052] In some implementations, the R 1 The heterocyclic alkyl group is selected from The heterocyclic alkyl group is optionally substituted with one or more R.

[0053] In some implementations, the R 1 Selected from -N(CH3)2, -N(CH2CH3)2, -N(CH3)(CH2CH3), OCH3,

[0054] In some other implementations, the R 1 Selected from -N(CH3)2, -N(CH2CH3)2, -N(CH3)(CH2CH3),

[0055] In some implementations, the R 2 Selected from hydrogen, halogen, hydroxyl, cyano, amino, nitro, C 1-7 Alkyl or C 1-7 Alkoxy, where C 1-7 Alkyl or C 1-7 The alkoxy group is optionally substituted with one or more R'. In some embodiments, the R' is... 2 Selected from hydrogen, halogen, hydroxyl, cyano, amino, nitro, C 1-6 Alkyl or C 1-6 Alkoxy, where C1-6 Alkyl or C 1-6 The alkoxy group is optionally substituted with one or more R'. In some embodiments, the R' is... 2 Selected from hydrogen, halogen, hydroxyl, cyano, amino, nitro, C 1-5 Alkyl or C 1-5 Alkoxy, where C 1-5 Alkyl or C 1-5 The alkoxy group is optionally substituted with one or more R'. In some embodiments, the R' is... 2 Selected from hydrogen, fluorine, chlorine, bromine, iodine, hydroxyl, cyano, amino, nitro, C 1-4 Alkyl or C 1-4 Alkoxy, where C 1-4 Alkyl or C 1-4 The alkoxy group is optionally substituted with one or more R'. In some embodiments, the R' is... 2 Selected from hydrogen, fluorine, chlorine, bromine, hydroxyl, cyano, amino, nitro, C 1-3 Alkyl or C 1-3 Alkoxy, where C 1-3 Alkyl or C 1-3 The alkoxy group is optionally substituted with one or more R'. In some embodiments, the R' is... 2 Selected from hydrogen, fluorine, chlorine, bromine or C 1-3 Alkyl, wherein C 1-3 The alkyl group is optionally substituted with one or more R'. In some embodiments, the R... 2 Selected from hydrogen, fluorine, chlorine or C 1-3 Alkyl, wherein C 1-3 The alkyl group is optionally substituted with one or more R'. In some embodiments, the R... 2 Selected from fluorine or C 1-3 Alkyl, wherein C 1-3 The alkyl group is optionally substituted with one or more fluorine molecules. In some embodiments, the R... 2 The methyl group is selected from fluorine or optionally substituted with one or more fluorine molecules. In some embodiments, the R... 2 Selected from fluorine, methyl, or trifluoromethyl.

[0056] In some implementations, the R 2 Selected from halogens. In some embodiments, the R... 2 Selected from fluorine. In some embodiments, the R... 2 Selected from C that is optionally substituted with one or more halogens 1-3 Alkyl group. In some embodiments, the R... 2 Selected from C 1-3 Alkyl group. In some embodiments, the R... 2 Selected from methyl. In some embodiments, the R...2 Selected from C that has been substituted with one or more fluorine molecules 1-3 Alkyl group. In some embodiments, the R... 2 Selected from trifluoromethyl.

[0057] In some embodiments, R' is selected from fluorine, chlorine, bromine, hydroxyl, cyano, amino, or nitro. In some embodiments, R' is selected from fluorine, chlorine, or bromine. In some embodiments, R' is selected from fluorine or chlorine. In some embodiments, R' is selected from fluorine.

[0058] In some implementations, the R 3 C is selected from hydrogen, fluorine, chlorine, bromine, or optionally substituted with one or more halogens. 1-6 Alkyl; in some embodiments, the R 3 C atoms selected from hydrogen, fluorine, chlorine, bromine, or optionally substituted by one or more atoms selected from fluorine or chlorine. 1-5 Alkyl; in some embodiments, the R 3 C atoms selected from hydrogen, fluorine, chlorine, bromine, or optionally substituted by one or more atoms selected from fluorine or chlorine. 1-4 Alkyl; in some embodiments, the R 3 C atoms selected from hydrogen, fluorine, chlorine, or optionally substituted with one or more fluorine atoms 1-3 Alkyl group. In some embodiments, the R... 3 The R is selected from hydrogen, fluorine, chlorine, or a methyl group optionally substituted with one or more fluorine atoms. In some embodiments, the R... 3 Selected from hydrogen, fluorine, chlorine, or trifluoromethyl.

[0059] In some implementations, the R 3 Selected from halogens. In some embodiments, the R... 3 Selected from C substituted with one or more halogens 1-3 Alkyl group. In some embodiments, the R... 3 Selected from hydrogen. In some embodiments, the R... 3 Selected from fluorine. In some embodiments, the R... 3 Selected from chlorine. In some embodiments, the R... 3 Selected from trifluoromethyl.

[0060] In some implementations, structural fragments Selected from

[0061] In other implementations, structural fragments Selected from

[0062] In some implementations, structural fragments Selected from

[0063] In other implementations, structural fragments Selected from

[0064] In some implementations, this application includes the variables defined above and their implementations, as well as any combination thereof.

[0065] In some embodiments, the compound of formula I of this application, or its pharmaceutically acceptable salt, stereoisomer, or tautomer, is selected from compounds of formula II, III, IV, V, or VI, or their pharmaceutically acceptable salts, stereoisomers, or tautomers.

[0066]

[0067] Among them, L and R 1 R 2 Or R 3 The definition is the same as in this application. In some embodiments, this application provides the following compounds or their pharmaceutically acceptable salts, stereoisomers, and tautomers:

[0068]

[0069]

[0070]

[0071] On the other hand, this application relates to pharmaceutical compositions comprising a compound of Formula I of this application or a pharmaceutically acceptable salt, stereoisomer, or tautomer thereof. In some embodiments, the pharmaceutical compositions of this application further include pharmaceutically acceptable excipients.

[0072] On the other hand, this application relates to a method for treating EGFR-mediated diseases in mammals, comprising administering to a mammal, preferably a human, a therapeutically effective amount of a compound of formula I or a pharmaceutically acceptable salt, stereoisomer, tautomer, or pharmaceutical composition thereof.

[0073] On the other hand, this application relates to the use of a compound of formula I or a pharmaceutically acceptable salt, stereoisomer, tautomer, or pharmaceutical composition thereof in the preparation of a medicament for the prevention or treatment of EGFR-mediated diseases.

[0074] On the other hand, this application relates to the use of a compound of formula I or a pharmaceutically acceptable salt, stereoisomer, tautomer, or pharmaceutical composition thereof in the prevention or treatment of EGFR-mediated diseases.

[0075] On the other hand, this application relates to Formula I compounds or pharmaceutically acceptable salts, stereoisomers, tautomers, or pharmaceutical compositions thereof for the prevention or treatment of EGFR-mediated diseases.

[0076] In some embodiments, the EGFR-mediated diseases described in this application are selected from diseases mediated by EGFR mutations. The mutations are selected from one, two, three, or four of L858R, T790M, d19, and C797S; in some embodiments, the mutations are selected from two mutations, L858R and T790M; in some embodiments, the mutations are selected from two mutations, d19 and T790M. Further, in some embodiments, the mutations include the C797S mutation; even further, the mutations are selected from three mutations, L858R, T790M, and C797S; or the mutations are selected from three mutations, d19, T790M, and C797S.

[0077] In some implementations, the EGFR-mediated diseases described in this application are selected from cancer.

[0078] In some implementations, the EGFR-mediated diseases described in this application are selected from lung cancer.

[0079] In some implementations, the EGFR-mediated disease described in this application is selected from non-small cell lung cancer.

[0080] The compounds of this application have good kinase and cellular activity (including wild-type and mutant types, such as d19, T790M, C797S and L858R), stable in vitro liver microsomal metabolism, in vivo pharmacokinetic data and efficacy.

[0081] definition

[0082] Unless otherwise stated, the following terms as used in this application shall have the following meanings. A particular term should not be considered uncertain or unclear unless specifically defined, but should be understood in accordance with its ordinary meaning in the art. When a trade name appears herein, it is intended to refer to the corresponding product or its active ingredient.

[0083] The term "substitution" refers to the substitution of one or more hydrogen atoms on a specific atom by a substituent, provided that the valence state of the specific atom is normal and the resulting compound is stable. When the substituent is oxo (i.e., =O), it means that two hydrogen atoms are substituted; oxo substitution does not occur on aromatic groups.

[0084] The terms “optional” or “optionally” mean that the event or condition subsequently described may or may not occur, including both the occurrence and non-occurrence of said event or condition. For example, the ethyl group “optionally” being halogenated means that the ethyl group can be unsubstituted (CH2CH3), monosubstituted (e.g., CH2CH2F), polysubstituted (e.g., CHFCH2F, CH2CHF2, etc.), or fully substituted (CF2CF3). Those skilled in the art will understand that for any group containing one or more substituents, no substitution or substitution pattern that is spatially impossible and / or cannot be synthesized is introduced.

[0085] C in this article m-n This means that the part has an integer number of carbon atoms within a given range. For example, "C 1-6 "" means that the group can have 1 carbon atom, 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms or 6 carbon atoms.

[0086] When any variable (e.g., R) appears more than once in the composition or structure of a compound, its definition is independent in each case. Therefore, for example, if a group is substituted by two Rs, each R has an independent option.

[0087] When the number of a linking group is 0, such as -(CH2)0-, it indicates that the linking group is a covalent single bond.

[0088] When one of the variables is selected as a single bond, it means that the two groups it connects to are directly connected, such as XLR. 1 When L represents a single bond, it indicates that the structure is actually XR. 1 When multiple consecutive variables are selected from single bonds, it indicates that the two groups they connect are directly linked, for example... When both X and L are selected from single bonds, it indicates that the structure is actually...

[0089] When a substituent is cross-bonded to two atoms on a ring, it can bond to any atom on that ring. For example, structural units. This indicates that it can be substituted at any position on the cyclohexyl group or cyclohexadiene.

[0090] This indicates that the substituent is attached to other structures. For example, when R... 1 The heterocyclic alkyl group is selected from When, it means Connected to L: The term "halogen" or "halogen" refers to fluorine, chlorine, bromine, and iodine.

[0091] The term "hydroxyl group" refers to the -OH group.

[0092] The term "cyano" refers to the -CN group.

[0093] The term "amino" refers to the -NH2 group.

[0094] The term "nitro" refers to the -NO2 group.

[0095] The term "alkyl" refers to a compound with the general formula C10. n H 2n+1 The alkyl group. This alkyl group can be straight-chain or branched. For example, the term "C 1-6 "Alkyl" refers to an alkyl group containing 1 to 6 carbon atoms (e.g., methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, 1-methylbutyl, 2-methylbutyl, 3-methylbutyl, neopentyl, hexyl, 2-methylpentyl, etc.). Similarly, the alkyl portion (i.e., alkyl) of alkoxy, alkylamino, dialkylamino, alkylsulfonyl, and alkylthio groups has the same definition as above.

[0096] The term "alkylene" refers to a divalent group formed by removing one hydrogen atom from any position of an alkyl group. For example, the term "C..." 1-6 "Alkylene" refers to an alkylene containing 1 to 6 carbon atoms, such as methylene (-CH2-), ethylene including but not limited to -CH2CH2-, propylene including but not limited to -CH2CH2CH2- and -CH(CH3)CH2-, and butylene including but not limited to -CH2CH2CH2CH2-.

[0097] The term "alkenyl" refers to an unsaturated aliphatic hydrocarbon group consisting of a straight or branched chain of carbon and hydrogen atoms, having at least one double bond. Non-limiting examples of alkenyl groups include, but are not limited to, vinyl, 1-propenyl, 2-propenyl, 1-butenyl, isobutenyl, 1,3-butadienyl, etc.

[0098] The term "alkenyl" refers to a divalent group formed by removing one hydrogen atom from any position of an alkenyl group. For example, the term "C..." 2-8 "Alkenyl" refers to alkenyl groups containing 2 to 8 carbon atoms, such as vinylene (-CH=CH-), and butylene, including but not limited to -CH2CH=CHCH2- and -CH2CH2CH=CH-.

[0099] The term "alkoxy" refers to -O-alkyl.

[0100] The term "alkylamino" refers to -NH-alkyl.

[0101] The term "dialkylamino" refers to -N(alkyl)2.

[0102] The term "alkylsulfonyl" refers to -SO2-alkyl.

[0103] The term "cycloalkyl" refers to a fully saturated carbon ring that may exist as a monocyclic, bridged, or spirocyclic ring. Unless otherwise indicated, the carbon ring is typically a 3- to 10-membered ring. Non-limiting examples of cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, norbornyl (bicyclo[2.2.1]heptyl), bicyclo[2.2.2]octyl, adamantyl, etc.

[0104] The term "heterocyclic alkyl" refers to a fully saturated cyclic group that may exist as a monocyclic, bridged, or spirocyclic ring. Unless otherwise indicated, the heterocycle is typically a ring containing one to three heteroatoms (preferably one or two heteroatoms) independently selected from sulfur, oxygen, and / or nitrogen. Examples of 3-membered heterocyclic alkyl groups include, but are not limited to, ethylene oxide, cyclothioethylene, and cycloazoethylene; non-limiting examples of 4-membered heterocyclic alkyl groups include, but are not limited to, acridine, oxadiazolyl, and thiobutyl; examples of 5-membered heterocyclic alkyl groups include, but are not limited to, tetrahydrofuranyl, tetrahydrothiophenyl, pyrrolidinyl, isoxazolyl, oxazolyl, isothiazolyl, thiazolyl, imidazolyl, and tetrahydropyrazolyl; examples of 6-membered heterocyclic alkyl groups include, but are not limited to, piperidinyl, tetrahydropyranyl, tetrahydrothiaranyl, morpholinyl, piperazine, 1,4-thiaoxane, 1,4-dioxane, thiomorpholinyl, 1,3-dithiaalkyl, and 1,4-dithiaalkyl; and examples of 7-membered heterocyclic alkyl groups include, but are not limited to, azirheptanyl, oxeheptanyl, and thioheptanyl. Monocyclic heterocyclic alkyl groups having 5 or 6 ring atoms are preferred.

[0105] The term "treatment" means administering the compound or preparation described in this application to improve or eliminate a disease or one or more symptoms related to said disease, and includes:

[0106] (i) Suppress the disease or disease state, that is, curb its development;

[0107] (ii) Relieve the disease or disease state, even if the disease or disease state subsides.

[0108] The term “prevention” means administering the compound or formulation described in this application to prevent a disease or one or more symptoms associated with the disease, and includes preventing the occurrence of a disease or disease state in mammals, particularly when such mammals are susceptible to the disease state but have not yet been diagnosed with the disease state.

[0109] The term "effective amount" means the amount of the compound of this application used to (i) treat or prevent a particular disease, condition, or disorder; (ii) alleviate, improve, or eliminate one or more symptoms of a particular disease, condition, or disorder; or (iii) prevent or delay the onset of one or more symptoms of a particular disease, condition, or disorder described herein. The amount of the compound of this application constituting an "effective amount" varies depending on the compound, the disease state and its severity, the route of administration, and the age of the mammal to be treated, but may routinely be determined by a person skilled in the art based on their own knowledge and the present disclosure.

[0110] The term "pharmaceutical acceptable" refers to compounds, materials, compositions, and / or dosage forms that, within the bounds of reliable medical judgment, are suitable for use in contact with human and animal tissues without excessive toxicity, irritation, allergic reactions, or other problems or complications, in proportion to a reasonable benefit / risk ratio.

[0111] As pharmaceutically acceptable salts, for example, metal salts, ammonium salts, salts formed with organic bases, salts formed with inorganic acids, salts formed with organic acids, and salts formed with basic or acidic amino acids may be mentioned.

[0112] The term "pharmaceutical composition" refers to a mixture of one or more compounds of this application or their salts with pharmaceutically acceptable excipients. The purpose of a pharmaceutical composition is to facilitate the administration of the compounds of this application to an organism.

[0113] The term "pharmaceuticalally acceptable excipient" refers to excipients that do not cause significant irritation to the organism and do not impair the biological activity and properties of the active compound. Suitable excipients are well known to those skilled in the art, such as carbohydrates, waxes, water-soluble and / or water-swellable polymers, hydrophilic or hydrophobic materials, gelatin, oils, solvents, water, etc.

[0114] The word “comprise” or “include” and its English variants such as comprises or comprising should be understood in an open, non-exclusive sense, meaning “including but not limited to”.

[0115] The compounds and intermediates of this application may also exist in different tautomer forms, and all such forms are included within the scope of this application. The terms "tautomer" or "tautomer form" refer to structural isomers of different energies that can interconvert via low-barrier transitions. For example, proton tautomers (also known as proton transfer tautomers) include interconversions via proton migration, such as keto-enol and imine-enamine isomerization. A specific example of a proton tautomer is the imidazole moiety, where a proton can migrate between two ring nitrogens. Valence tautomers include interconversions via the recombination of some bonding electrons. Non-limiting examples of tautomers include, but are not limited to, those shown below.

[0116] This application also includes compounds of this application that are identical to those described herein, but with one or more atoms replaced by isotopes whose atomic weights or mass numbers differ from those commonly found in nature. Examples of isotopes that can be incorporated into compounds of this application include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, iodine, and chlorine, such as... 2 H, 3 H, 11 C 13 C 14 C 13 N、 15 N、 15 O、 17 O、 18 O、 31 P, 32 P, 35 S, 18 F, 123 I, 125 I and 36 Cl, etc.

[0117] Certain isotope-labeled compounds of this application (e.g., using...) 3 H and 14 Those labeled with C can be used in the analysis of compound and / or substrate tissue distribution. Tritiumization (i.e. 3 H) and carbon-14 (i.e. 14 C) Isotopes are particularly preferred due to their ease of preparation and detectability. Positron-emitting isotopes, such as... 15 O、 13 N、 11 C and 18 F can be used in positron emission tomography (PET) studies to determine substrate occupancy. The isotopically labeled compounds of this application can typically be prepared by replacing the unlabeled reagent with an isotopically labeled reagent using a procedure similar to those disclosed in the schemes and / or examples below.

[0118] In addition, heavier isotopes (such as deuterium) are used. 2 H)) substitution can provide certain therapeutic advantages resulting from higher metabolic stability (e.g., increased in vivo half-life or reduced dose requirement), and may therefore be preferred in certain situations, where deuterium substitution can be partial or complete, with partial deuterium substitution referring to at least one hydrogen being replaced by at least one deuterium.

[0119] The compounds of this application may be asymmetric, for example, having one or more stereoisomers. Unless otherwise stated, all stereoisomers include, such as enantiomers and diastereomers. The compounds containing asymmetric carbon atoms of this application can be isolated in optically active pure form or in racemic form. The optically active pure form can be resolved from a racemic mixture or synthesized using chiral starting materials or chiral reagents.

[0120] The pharmaceutical compositions of this application can be prepared by combining the compounds of this application with suitable pharmaceutically acceptable excipients, for example, in solid, semi-solid, liquid or gaseous formulations, such as tablets, pills, capsules, powders, granules, ointments, emulsions, suspensions, suppositories, injections, inhalers, gels, microspheres and aerosols.

[0121] Typical routes of administration for the compounds of this application or their pharmaceutically acceptable salts or pharmaceutical compositions thereof include, but are not limited to, oral, rectal, topical, inhalation, parenteral, sublingual, vaginal, intranasal, intraocular, intraperitoneal, intramuscular, subcutaneous, and intravenous administration.

[0122] The pharmaceutical composition of this application can be manufactured using methods well known in the art, such as conventional mixing, dissolving, granulation, sugar-coated pill making, grinding, emulsification, freeze drying, etc.

[0123] In some embodiments, the pharmaceutical composition is in oral form. For oral administration, the pharmaceutical composition can be formulated by mixing the active compound with pharmaceutically acceptable excipients well known in the art. These excipients enable the compounds of this application to be formulated into tablets, pills, lozenges, sugar-coated tablets, capsules, liquids, gels, pastes, suspensions, etc., for oral administration to patients.

[0124] Solid oral compositions can be prepared using conventional mixing, filling, or tableting methods. For example, they can be obtained by mixing the active compound with solid excipients, optionally milling the resulting mixture, adding other suitable excipients if necessary, and then processing the mixture into granules to obtain the core of a tablet or sugar-coated formulation. Suitable excipients include, but are not limited to, binders, diluents, disintegrants, lubricants, glidants, sweeteners, or flavoring agents.

[0125] The pharmaceutical composition may also be suitable for parenteral administration, such as in suitable unit dosage forms of sterile solutions, suspensions or lyophilized products.

[0126] In all methods of administration of the compounds of general formula I described herein, the daily dose is from 0.01 to 200 mg / kg body weight, in the form of single or separate doses.

[0127] The compounds of this application can be prepared by a variety of synthetic methods known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combining them with other chemical synthetic methods, and equivalent substitutions known to those skilled in the art. Preferred embodiments include, but are not limited to, the embodiments of this application.

[0128] The chemical reactions in the specific embodiments of this application are carried out in a suitable solvent, which must be suitable for the chemical changes and the reagents and materials required in this application. In order to obtain the compounds of this application, it is sometimes necessary for those skilled in the art to modify or select the synthesis steps or reaction process based on existing embodiments.

[0129] An important consideration in synthetic route planning in this field is the selection of appropriate protecting groups for reactive functional groups. For example, see Greene's Protective Groups in Organic Synthesis (4th Ed). Hoboken, New Jersey: John Wiley & Sons, Inc. All references cited in this application are incorporated herein by reference in their entirety.

[0130] This application uses the following abbreviations:

[0131] Pd(dppf)CH2Cl2 is the [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride dichloromethane complex; PdCl2(dppf) is the [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride; DMF represents N,N-dimethylformamide; TBTU represents O-benzotriazole-N,N,N',N'-tetramethylurea tetrafluoroboronic acid; DPPB represents 1,4-bis(diphenylphosphine)butane; Boc represents tert-butoxycarbonyl; TBAB represents tetrabutylammonium bromide; DMF represents N,N-dimethylformamide; NMI represents N-methylimidazolium; TCFH represents tetramethylchlorourea hexafluorophosphate. Detailed Implementation

[0132] For clarity, this application is further illustrated with examples, but these examples are not intended to limit the scope of this application. All reagents used in this application are commercially available and can be used without further purification.

[0133] Example 1:

[0134]

[0135]

[0136] (1) Preparation method of compound A:

[0137] 2-Chloro-6-methylisonicotinic acid methyl ester (20.0 g), 1-methyl-5-hydroxypyrazole (21.4 g), Pd(dppf)CH2Cl2 (2.64 g), and sodium carbonate (25.125 g) were dissolved in anisole (400 mL). The reaction solution was purged with argon three times and stirred at 130 °C for approximately 10 h. After the reaction was complete, the solution was cooled to room temperature, filtered, and the filter cake was rinsed with 80 mL of toluene. 60 mL of methanol was added to the filtrate, followed by the slow addition of 40 mL of 4M dioxane chloride solution. After the addition was complete, the solution was stirred at room temperature for 2 h. After stirring, the solution was filtered, and the filter cake was rinsed with 40 mL of toluene. The target compound was obtained and dried under vacuum at 50 °C for 8 h. Compound A was obtained, ESI-MS: m / z = 248.26 [M+H] + .

[0138] (2) Preparation method of compound B:

[0139] 2-Fluoro-4-bromonitrobenzene (9.17 g), (R)-5-amino-4-methyl-1-pentanol (43.78 mmol), and potassium carbonate (12.68 g) were dissolved in 200 mL of DMF and stirred at room temperature for 4 h until the reaction was complete. The mixture was filtered, and the filter cake was washed with 40 mL of methyl tert-butyl ether. The filtrate was concentrated at 40 °C until no liquid flowed out, yielding the target product B. No further purification was required, and it was used directly in the next reaction. ESI-MS: m / z = 317.18 [M+H] + .

[0140] (3) Preparation method of compound C:

[0141] Compound B (41.7 mmol) was dissolved in 300 mL of dichloromethane and stirred at room temperature. Triethylamine (6.33 g) was slowly added, and after the addition was complete, the reaction flask was placed in an ice-water bath and stirred for 10 min. Then, methanesulfonyl chloride (5.25 g) was slowly added dropwise. After the addition was complete, the reaction flask was placed in an ice-water bath and stirred for about 2 h until the reaction was complete. The mixture was filtered, and the filter cake was washed with 40 mL of methyl tert-butyl ether. The filtrate was extracted with methyl tert-butyl ether (50 mL x 3) and purified water (50 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated until no liquid flowed out to give compound C. No further purification was required, and it was used directly in the next reaction. ESI-MS: m / z = 395.03 [M+H] + .

[0142] (4) Preparation method of compound D:

[0143] Compound A (11.8 g), compound C (41.7 mmol), and potassium carbonate (14.4 g) were dissolved in 400 mL of DMF and reacted with stirring at 60 °C for approximately 12 h until the reaction was complete. The mixture was filtered, and the filter cake was washed with 40 mL of methyl tert-butyl ether. The filtrate was washed with purified water (50 mL x 3), and the aqueous phase was extracted with methyl tert-butyl ether (50 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated until no liquid eluent was observed. Column chromatography was used to obtain compound D. ESI-MS: m / z = 546.16 [M+H] + .

[0144] (5) Preparation method of compound E:

[0145] Compound D (5.0 g) was dissolved in 300 mL of methanol and stirred at room temperature for 20 min to ensure complete dispersion. Raney Ni was slowly added, and the reaction mixture was purged with nitrogen three times, then with hydrogen twice, while stirring under hydrogen atmosphere. The reaction proceeded to completion. The mixture was filtered, and the filter cake was washed with 50 mL of methanol. The filtrate was collected and concentrated at 40 °C until no liquid flowed out, yielding compound E. ESI-MS: m / z = 516.36 [M+H] + .

[0146] (6) Preparation method of compound F:

[0147] Compound E (8.0 g) and cyanogen bromide (3.3 g) were dissolved in 200 mL of dichloromethane and 200 mL of tert-butanol. The reaction was stirred at room temperature for 18 h. After the reaction was complete, the mixture was washed with purified water (200 mL), retaining the organic phase, and the aqueous phase was extracted with 50 mL of dichloromethane. The organic phases were combined and dried over anhydrous sodium sulfate. The solution was concentrated at 35 °C until no liquid eludes, yielding compound F. ESI-MS: m / z = 541.28 [M+H] + .

[0148] (7) Preparation method of compound G:

[0149] Compound F (8.3 g) was dissolved in 166 mL of tetrahydrofuran solution, and sodium hydroxide (2.45 g) was dissolved in 166 mL of purified water. The prepared sodium hydroxide aqueous solution was slowly added dropwise to the tetrahydrofuran solution of compound F. After the addition was complete, the reaction was stirred at room temperature for 4 h. After the reaction was complete, it was concentrated at 45 °C until no liquid flowed out. 6 M hydrochloric acid solution was added dropwise to the remaining reaction solution until the pH was adjusted to about 4-5. After the addition was complete, the mixture was stirred at room temperature for 1 h. The mixture was filtered, and the filter cake was washed with n-hexane to obtain compound G. ESI-MS: m / z = 527.20 [M+H] + .

[0150] (8) Preparation method of compound H:

[0151] Compound G (7.9 g) was dissolved in 300 mL of dichloromethane, and triethylamine (6.06 g) was slowly added dropwise. Then TBTU (5.8 g) was added, and the reaction was stirred at room temperature for 4 h. After the reaction was complete, the reaction solution was washed with 200 mL of purified water, retaining the organic phase. The aqueous phase was extracted with 50 mL of dichloromethane, retaining the organic phase. The organic phase was washed with purified water (20 mL * 3) and dried over anhydrous sodium sulfate. The solution was concentrated until no liquid flowed out, yielding compound H. ESI-MS: m / z = 509.3 [M + H] + .

[0152] (9) Preparation method of Example 1:

[0153] Compound H (100 mg), copper hydroxide (40 mg), and potassium carbonate (54.3 mg) were dissolved in 1.5 mL of dimethylaminoethanol. The reaction was carried out at 130 °C for 18 h. The reaction was complete. The mixture was extracted with purified water (10 mL x 3) and ethyl acetate (10 mL x 3). The organic phase was dried over anhydrous sodium sulfate. The mixture was concentrated under reduced pressure at 50 °C until no liquid flowed out. The product was purified by preparative liquid chromatography to obtain Example 1. ESI-MS: m / z = 518.42 [M+H] + .

[0154] Example 2:

[0155]

[0156] Compound H (100 mg), copper hydroxide (40 mg), and potassium carbonate (54.3 mg) were dissolved in 1.5 mL of N-(2-hydroxyethyl)-pyrrolidine. The reaction was carried out at 130 °C for 18 h. The reaction was complete. The mixture was extracted with purified water (10 mL x 3) and ethyl acetate (10 mL x 3). The organic phase was dried over anhydrous sodium sulfate. The mixture was concentrated under reduced pressure at 50 °C until no liquid flowed out. The solution was purified by preparative liquid chromatography to obtain Example 2. ESI-MS: m / z = 544.38 [M+H] + .

[0157] Example 3:

[0158]

[0159] Compound H (100 mg), copper hydroxide (40 mg), and potassium carbonate (54.3 mg) were dissolved in 1.5 mL of 3-dimethylamino-1-propanol. The reaction was carried out at 130 °C for 18 h. The reaction was complete. The mixture was extracted with purified water (10 mL x 3) and ethyl acetate (10 mL x 3). The organic phase was dried over anhydrous sodium sulfate. The mixture was concentrated under reduced pressure at 50 °C until no liquid eluent was observed. The solution was purified by preparative liquid chromatography to obtain Example 3. ESI-MS: m / z = 532.39 [M+H] + .

[0160] Example 4:

[0161]

[0162] Compound H (100 mg), copper hydroxide (40 mg), and potassium carbonate (54.3 mg) were dissolved in 1.5 mL of diethylaminoethanol. The reaction was carried out at 130 °C for 18 h. The reaction was complete. The mixture was extracted with purified water (10 mL x 3) and ethyl acetate (10 mL x 3). The organic phase was dried over anhydrous sodium sulfate. The mixture was concentrated under reduced pressure at 50 °C until no liquid flowed out. The product was purified by preparative liquid chromatography to obtain Example 4. ESI-MS: m / z = 546.41 [M+H] + .

[0163] Example 5:

[0164]

[0165] Compound H (50 mg), 1-methylpiperidin-4-amine (114 mg), 2-(di-tert-butylphosphine)biphenyl (11.9 mg), tris(dibenzylacetone)dipalladium (18.3 mg), and potassium tert-butoxide (112.5 mg) were sequentially added to tert-amyl alcohol (3 mL). The mixture was purged three times with argon gas, and the reaction was carried out at 115 °C for 2.5 h. The filtrate and concentrate were purified by preparative liquid chromatography to obtain Example 5. ESI-MS: m / z = 543.45 [M+H] + .

[0166] 1H NMR (500MHz, DMSO-d6) δ8.41(s,1H),7.92(s,1H),7.55(s,1H),7.24(d,J=8.6Hz,1H),6.67(d,J=1.1Hz,1 H),6.55(dd,J=8.6,1.6Hz,1H),5.44(s,1H),4.36(td,J=8.9,4.6Hz,1H),4.22–4.08(m,1H),4.03–3.92( m,1H),3.83(s,1H),3.73(s,3H),3.27(t,J=9.8Hz,1H),2.86–2.68(m,3H),2.55(s,3H),2.19(s,3H),2.0 8(t,J=11.1Hz,2H),1.94–1.90(m,2H),1.87(s,3H),1.40(dt,J=13.1,10.4Hz,3H),0.82(d,J=6.5Hz,3H).

[0167] Example 6:

[0168]

[0169] Compound H (50 mg), 3-dimethylamino-1-propylamine (102 mg), 2-(di-tert-butylphosphine)biphenyl (11.9 mg), tris(dibenzylacetone)dipalladium (18.3 mg), and potassium tert-butoxide (112.5 mg) were sequentially added to tert-amyl alcohol (3 mL). The mixture was purged three times with argon gas, and the reaction was carried out at 115 °C for 2.5 h. The residue was filtered, concentrated, and purified by preparative liquid chromatography to obtain Example 6 (13 mg). ESI-MS: m / z = 531.45 [M+H] + .

[0170] 1H NMR (500MHz, DMSO-d6) δ8.41(s,1H),7.92(s,1H),7.49(d,J=61.6Hz,1H),7.24(t,J=9.7Hz,1H),6.63(d,J=1.7Hz,1 H),6.53(dd,J=8.6,1.9Hz,1H),4.36(td,J=9.1,4.6Hz,1H),4.14(dd,J=13.6,2.8Hz,1H),4.05–3.91(m,1H),3.81( dd,J=13.5,10.4Hz,1H),3.73(s,3H),3.08(t,J=6.9Hz,2H),2.78(d,J=4.0Hz,1H),2.55(s,3H),2.34(t,J=7.1Hz,2 H),2.25–2.19(m,1H),2.16(s,6H),2.01–1.89(m,2H),1.79–1.64(m,2H),1.52–1.37(m,1H),0.82(d,J=6.5Hz,3H).

[0171] Example 7:

[0172]

[0173] (1) Preparation method of compound I:

[0174] Compound H (300 mg), N-formylsaccharin (249 mg), palladium acetate (13.2 mg), DPPB (37.7 mg), and sodium carbonate (125 mg) were added to a reaction tube. The mixture was purged three times with N2. While maintaining the N2 atmosphere, a mixed solution of triethylsilane (119 mg) and DMF (10 mL) was added. The mixture was stirred under sealed conditions at room temperature for 20 min, then stirred at 80 °C for 16 h. The reaction solution was concentrated under reduced pressure at 70 °C, and 20 mL of ethyl acetate was added and stirred for 20 min. The mixture was filtered, and the filtrate was concentrated under reduced pressure to obtain compound I. ESI-MS: m / z = 459.4 [M+H] + .

[0175] (2) Preparation method of Example 7:

[0176] Compound I (20 mg), piperidine (5.7 mg), dichloromethane (1.5 mL), acetic acid (95.2 mg), and sodium triacetoxyborohydride (18.5 mg) were added sequentially to a flask and stirred at room temperature for 23 h. The reaction solution was filtered, the filtrate was concentrated under reduced pressure, and purified by preparative liquid chromatography to obtain Example 7. ESI-MS: m / z = 528.43 [M+H] + .

[0177] Example 8:

[0178]

[0179] (1) Preparation of compound J:

[0180] 20.0 g of 3-fluoro-4-nitrobenzoic acid was dispersed in 400 mL of dichloromethane. Oxaloyl chloride (15.1 g) and DMF (0.4 g) were added dropwise at 25 °C, and the mixture was stirred at 25 °C for 1 h. After the system was dissolved, the temperature was lowered to 5-10 °C, and 13.0 g of N-methylpiperazine was added dropwise. The reaction was carried out at 5-10 °C for 2 h until the reaction was complete. 12 g of sodium carbonate was dissolved in 200 mL of purified water. 200 mL of sodium carbonate aqueous solution was added to the system. The mixture was separated, and the dichloromethane phase was washed sequentially with 300 mL of water and 200 mL of 10% sodium chloride solution. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure until no liquid flowed out, yielding compound J.

[0181] (2) Preparation method of compound B-1:

[0182] Compound J (2.14 g), (R)-5-amino-4-methyl-1-pentanol (8.00 mmol, 1.0 eq), and potassium carbonate (1.66 g) were dissolved in 15 mL of DMF and stirred at room temperature for 6 h until the reaction was complete. 200 mL of water was added to the reaction mixture, and the mixture was extracted with ethyl acetate (100 mL x 4). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and evaporated under reduced pressure to obtain the target product B-1. No further purification was required; this product was used directly in the next reaction.

[0183] (2) Preparation method of compound C-1:

[0184] Compound B-1 (8.00 mmol) was dissolved in 30 mL of dichloromethane, and triethylamine (1.62 g) was added. The mixture was cooled to 0–10 °C in an ice-water bath, and methanesulfonyl chloride (1.83 g) was slowly added dropwise. After the addition was complete, the reaction flask was placed at room temperature and stirred for about 2 h until the reaction was complete. The reaction solution was extracted with saturated sodium bicarbonate solution, and the organic phase was dried over anhydrous sodium sulfate. The mixture was filtered, and the filtrate was evaporated to dryness under reduced pressure to give compound C-1 (8.00 mmol), which could be used directly in the next reaction without further purification.

[0185] (3) Preparation method of compound D-1:

[0186] Compound C-1 (8.00 mmol), compound A (1.82 g), and potassium carbonate (2.76 g) were dissolved in 30 mL of DMF. The mixture was stirred at 60–70 °C for approximately 12 h until the reaction was complete. 200 mL of water was added to the reaction solution, and the mixture was extracted with ethyl acetate (200 mL x 2). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure until no liquid eluent was observed. Compound D-1 was purified by column chromatography. ESI-MS: m / z = 594.35 [M+H] + .

[0187] (4) Preparation method of compound E-1:

[0188] Compound D-1 (2.1 g) was dissolved in 40 mL of methanol. The reaction solution was purged with nitrogen three times. 10% palladium on carbon (0.2 g, dry basis) was added, and the solution was purged with hydrogen three times. The reaction was allowed to proceed at room temperature for 3.5 h until complete. After purging with nitrogen three times, the mixture was filtered. The filtrate was evaporated under reduced pressure until no liquid flowed out, yielding compound E-1, which was directly added to the next step. ESI-MS: m / z = 564.4 [M+H] + .

[0189] (5) Preparation method of compound F-1:

[0190] Compound E-1 (1.5 g) was dissolved in 30 mL of methanol, and 0.3 g of trifluoroacetic acid was added. Then, 2.0 mL of acetonitrile solution containing 0.34 g of cyanogen bromide was added dropwise at room temperature. The reaction was allowed to proceed overnight at room temperature until complete. The reaction solution was poured into 10 mL of saturated sodium bicarbonate solution, evaporated to dryness under reduced pressure, and the concentrate was extracted with dichloromethane (100 mL * 2). The organic phase was dried over sodium sulfate, evaporated to dryness under reduced pressure, and purified by column chromatography to give compound F-1. ESI-MS: m / z = 589.3 [M + H] + .

[0191] (6) Preparation method of compound G-1:

[0192] Compound F-1 (0.8 g) was dissolved in 20 mL of tetrahydrofuran solution, and sodium hydroxide (0.22 g) was dissolved in 20 mL of purified water. The prepared sodium hydroxide aqueous solution was slowly added dropwise to the tetrahydrofuran solution of compound F-1. The reaction was carried out at room temperature for 2 h. The tetrahydrofuran was removed by evaporation under reduced pressure. The pH was adjusted to 5-6 with 1 mol / L hydrochloric acid, and the water was removed by evaporation under reduced pressure to obtain compound G-1, which was directly added to the next step.

[0193] (7) Preparation method of Example 8:

[0194] Compound G-1 (1.36 mmol) was dissolved in 40 mL of dichloromethane, and triethylamine (0.55 g) was added. The mixture was dried over anhydrous sodium sulfate for 1 h, filtered, and the filtrate was treated with TBTU (0.52 g). The reaction was allowed to proceed overnight at room temperature. After the reaction was complete, the mixture was washed with 100 mL of purified water. The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated until no liquid flowed out. The solution was purified by column chromatography to obtain Example 8. ESI-MS: m / z = 557.36 [M+H] + .

[0195] Example 9:

[0196]

[0197] (1) Preparation method of compound K

[0198] Compound I (50 mg), N-Boc piperazine (30.5 mg), acetic acid (13.1 mg), sodium triacetoxyborohydride (46.2 mg), and dichloromethane (5 mL) were added to a flask and stirred at room temperature for 24 h. 5 mL of water was added, and the mixture was stirred and separated. The pH of the aqueous phase was adjusted to 9–10 with saturated sodium hydroxide solution, and then extracted with 5 mL of dichloromethane. The organic phases were separated, combined, and concentrated under reduced pressure to obtain compound K, which was directly added to the next step.

[0199] (2) Preparation method of Example 9

[0200] 2 mL of ethyl hydrogen chloride solution (4 mol / L) was added to compound K (obtained in the previous step), and the mixture was stirred at room temperature for 24 h. The reaction solution was concentrated under reduced pressure, and the concentrate was purified by preparative liquid chromatography to obtain Example 9. ESI-MS: m / z = 529.4 [M+H] + .

[0201] Example 10:

[0202]

[0203] (1) Preparation method of 3-methylenepyrrolidine hydrochloride

[0204] 9.5 g of 3-methylenepyrrolidine-1-carboxylic acid tert-butyl ester, 70 mL of dioxane, and 22 mL of concentrated hydrochloric acid were added to a flask and stirred at room temperature for 12 h. The reaction solution was concentrated under reduced pressure to obtain 3-methylenepyrrolidine hydrochloride, which was directly added to the next step.

[0205] (2) Preparation method of compound L

[0206] 3-Methylenepyrrolidine hydrochloride (310 mg) was dissolved in 10 mL of dichloromethane. Cyclopropane (368 mg), acetic acid (315 mg), and sodium triacetoxyborohydride (1.1 g) were added sequentially, and the mixture was stirred at room temperature for 12 h. 15 mL of water was added to the reaction mixture, and the mixture was stirred and separated, collecting the organic phase. The pH of the aqueous phase was adjusted to 9–10 by adding saturated sodium hydroxide solution dropwise. The mixture was extracted three times with dichloromethane, and the organic phases were combined and dried over anhydrous sodium sulfate for 5 h. The mixture was filtered, and the filtrate was concentrated under reduced pressure to give compound L. No purification was required; the compound was directly added to the next step. ESI-MS: m / z = 138.2 [M+H] + .

[0207] (3) Preparation method of Example 10

[0208] Compound H (170 mg) was dissolved in DMF (6 ml), and PdCl2 (dppf) (24.4 mg), TBAB (130 mg), potassium carbonate (138 mg), and compound L (138 mg) were added successively. The reaction solution was purged three times with argon gas, and stirred at 110 °C for 9 h under an argon atmosphere. The reaction solution was concentrated under reduced pressure until no liquid flowed out. The concentrate was purified by preparative liquid chromatography to obtain Example 10, ESI-MS: m / z = 566.5 [M+H]. + .

[0209] Example 11:

[0210]

[0211] Preparation method of Example 11:

[0212] Compound I (35 mg) was dissolved in dichloromethane (1.5 mL), and morpholine (10 mg), acetic acid (9.2 mg), and sodium triacetoxyborohydride (33 mg) were added. The mixture was stirred at room temperature for 12 h. 100 μL of water was added to the reaction solution, and the concentrate was obtained under reduced pressure. The concentrate was purified by preparative liquid chromatography to obtain Example 11. ESI-MS: m / z = 530.4 [M+H] + .

[0213] Example 12:

[0214]

[0215] (1) Preparation method of compound (3-bromopropyl)triphenylphosphine:

[0216] 1,3-Dibromopropane (90 mmol), triphenylphosphine (25 g), and toluene (260 mL) were added sequentially to a reaction flask, and the mixture was heated to 110 °C and reacted for 4 h. The reaction solution was filtered, and the filter cake was washed with 40 mL of cyclohexane to obtain compound (3-bromopropyl)triphenylphosphine. No further purification was required; it was used directly in the next reaction. ESI-MS: m / z = 383.01 [M] + .

[0217] (2) Preparation method of compound M:

[0218] Compound (3-bromopropyl)triphenylphosphine (5 g), methanol (26 mL), and dimethylamine (40 mmol) were added sequentially to a reaction flask. The mixture was stirred at room temperature for 1 h, then heated to 65 °C and reacted for 4 h. The reaction solution was concentrated until no liquid flowed out, yielding compound M. No further purification was required; it was used directly in the next reaction. ESI-MS: m / z = 348.4 [M] + .

[0219] (3) Preparation method of Example 12:

[0220] Compound I (20 mg), compound M (30 mg), dichloromethane (1.5 mL), acetic acid (95.2 mg), and sodium triacetoxyborohydride (18.5 mg) were added sequentially to a flask and stirred at room temperature for 23 h. The reaction solution was filtered, and the filtrate was concentrated under reduced pressure and purified by preparative liquid chromatography to obtain Example 12. ESI-MS: m / z = 528.39 [M+H] + .

[0221] 1 H NMR(500MHz,DMSO-d6)δ8.42(s,1H),8.25(s,1H),7.93(s,1H),7.65(s,1H),7.56(s,1H),7.46(d,J =7.9Hz,1H),7.26(d,J=7.8Hz,1H),6.56(d,J=15.7Hz,1H),6.42–6.23(m,1H),4.35(s,1H),4.17(d, J=12.7Hz,1H),4.03–3.95(m,2H),3.73(s,3H),2.81(s,1H),2.69(d,J=6.6Hz,2H),2.56(s,3H),2.4 1(s,6H),2.21(s,1H),2.06–1.82(m,3H),1.45(s,1H),1.18(t,J=6.8Hz,1H),0.82(d,J=5.5Hz,3H).

[0222] Example 13:

[0223]

[0224] Example 12 (50 mg), palladium hydroxide (5 mg), and methanol (5 mL) were added sequentially to a flask, purged three times with hydrogen, stirred at room temperature for 0.5 h, filtered, concentrated under reduced pressure, and purified by preparative liquid chromatography to obtain Example 13. ESI-MS: m / z = 530.42 [M+H] + .

[0225] 1 H NMR(500MHz,DMSO-d6)δ8.42(s,1H),8.23(s,1H),7.92(s,1H),7.56(s,1H),7.44(d,J=7.0Hz,2 H),7.07(d,J=8.1Hz,1H),4.36(d,J=4.1Hz,1H),4.18(d,J=12.0Hz,1H),3.99(s,1H),3.92(s,1H ),3.73(s,3H),2.80(s,1H),2.70(d,J=6.3Hz,2H),2.56(s,3H),2.36(s,6H),2.22(s,1H),1.95( dd,J=27.3,12.3Hz,2H),1.63(dd,J=12.9,6.6Hz,2H),1.57–1.37(m,3H),0.81(d,J=6.2Hz,3H).

[0226] Example 14:

[0227]

[0228] (1) Preparation method of compound 1-Boc-3-methylenepiperidine:

[0229] Methyltriphenylphosphine bromide (53 g) and tetrahydrofuran (150 mL) were added sequentially to the reaction flask, purged with nitrogen three times, and then a solution of potassium tert-butoxide in tetrahydrofuran (150 mmol) was slowly added. After the addition was complete, the reaction flask was placed in an ice-water bath and stirred for 20 min. N-tert-butoxycarbonyl-4-piperidinone (25 g) was added, and the reaction was carried out at room temperature for 4 h. The mixture was filtered, and the filtrate was slurried with 100 mL of n-heptane, resulting in the precipitation of a solid. The solid was filtered again, and the filter cake was washed with 40 mL of n-heptane to obtain compound 1-Boc-3-methylenepiperidine, which could be used directly in the next reaction without further purification.

[0230] (2) Preparation method of compound 3-methylenepiperidine hydrochloride:

[0231] 1-Boc-3-methylenepiperidine (12 g) and methyl tert-butyl ether (20 mL) were added sequentially to a single-necked flask, and ethyl hydrogen chloride solution (40 mmol, 10 mL) was added dropwise. The mixture was stirred at room temperature for 0.5 h, filtered, and the filter cake was washed with 40 mL of methyl tert-butyl ether to obtain 3-methylenepiperidine hydrochloride, which could be used directly in the next reaction without further purification.

[0232] (3) Preparation method of compound 1-methyl-3-methylenepiperidine:

[0233] Compound 3-methylenepiperidine hydrochloride (19.3 mmol), formaldehyde (38.6 mmol), acetic acid (38.6 mmol), dichloromethane (20 mL), and sodium triacetoxyborohydride (38.6 mmol) were added sequentially to a flask and stirred at room temperature for 2 h. The reaction solution was filtered, and the filtrate was concentrated under reduced pressure at 20 °C to obtain 1-methyl-3-methylenepiperidine, which could be used directly in the next reaction without further purification.

[0234] (4) Preparation method of Example 14:

[0235] Compound H (200 mg) was dissolved in 6 mL of DMF. PdCl2 (dppf) (20.1 mg), TBAB (111 mg), potassium carbonate (117 mg), and 1-methyl-3-methylenepiperidine (52.3 mg) were added sequentially. The reaction mixture was purged three times with argon gas and stirred at 110 °C for 9 h under an argon atmosphere. The reaction mixture was concentrated under reduced pressure until no liquid flowed out. The concentrate was purified by preparative liquid chromatography to obtain Example 14. ESI-MS: m / z = 540.39 [M+H] + .

[0236] Example 15:

[0237]

[0238] Example 14 (50 mg), palladium hydroxide (5 mg), and methanol (5 mL) were added sequentially to a reaction flask. The mixture was purged with hydrogen three times, stirred at room temperature for 0.5 h, filtered, and the filtrate was concentrated under reduced pressure. The resulting solution was purified by preparative liquid chromatography to obtain Example 15. ESI-MS: m / z = 542.54 [M+H]. + .

[0239] Example 16:

[0240]

[0241] (1) Preparation method of compound L-1

[0242] 3-Methylenepyrrolidine hydrochloride (3 g) was dissolved in 100 mL of dichloromethane. Then, 37% formaldehyde aqueous solution (4.12 g), acetic acid (3.1 g), and sodium triacetoxyborohydride (10.8 g) were added sequentially, and the mixture was stirred at room temperature for 12 h. The reaction solution was filtered, and the filtrate was concentrated under reduced pressure to obtain compound L-1. No purification was required; it was directly added to the next step.

[0243] (2) Preparation method of Example 16

[0244] Compound H (100 mg) was dissolved in 6.5 mL of LDM, and PdCl2 (dppf) (43 mg), TBAB (76 mg), potassium carbonate (108 mg), and compound L-1 (65 mg) were added successively. The reaction solution was purged three times with argon gas, and stirred at 110 °C for 12 h under an argon atmosphere. The reaction solution was concentrated under reduced pressure until no liquid flowed out. The concentrate was purified by preparative liquid chromatography to obtain Example 16, ESI-MS: m / z = 526.41 [M+H]. + .

[0245] Example 17:

[0246]

[0247] Compound H (50 mg), N,N-dimethylethylenediamine (88 mg), 2-(di-tert-butylphosphine)biphenyl (11.9 mg), tris(dibenzylacetone)dipalladium (18.3 mg), and potassium tert-butoxide (112.5 mg) were sequentially added to tert-amyl alcohol (3 mL). The mixture was purged three times with argon gas, and the reaction was carried out at 115 °C for 2.5 h. The mixture was filtered, and the filtrate was concentrated under reduced pressure. The concentrate was purified by preparative liquid chromatography to obtain Example 17. ESI-MS: m / z = 517.39 [M+H] + .

[0248] 1H NMR (500MHz, DMSO-d6) δ12.40(s,1H),8.41(s,1H),7.92(s,1H),7.55(s,1H),7.25(d,J=8.6Hz,1H),6.71(d, J=1.6Hz,1H),6.57(dd,J=8.6,1.9Hz,1H),5.39(s,1H),4.36(td,J=9.1,4.5Hz,1H),4.15(dd,J=13.5,2.6Hz, 1H),4.01–3.97(m,1H),3.82(dd,J=13.5,10.4Hz,2H),3.73(s,3H),3.14(t,J=6.4Hz,2H),2.79(s,1H),2.55 (s,2H),2.48(t,J=6.6Hz,2H),2.21(s,6H),1.90(s,3H),1.45(dd,J=16.4,9.6Hz,1H),0.83(d,J=6.5Hz,3H).

[0249] Example 18

[0250]

[0251] Compound H (100 mg), copper hydroxide (40 mg), and potassium carbonate (54.3 mg) were dissolved in 1.5 mL of ethylene glycol monomethyl ether. The mixture was placed in a microwave reactor and reacted at 150 °C for 8 h until the reaction was complete. The mixture was extracted with purified water (10 mL x 3) and ethyl acetate (10 mL x 3). The organic phase was dried over anhydrous sodium sulfate. The mixture was concentrated under reduced pressure at 50 °C until no liquid eluent was observed. The solution was purified by preparative liquid chromatography to obtain Example 18. ESI-MS: m / z = 505.60 [M+H] + .

[0252] Example 19

[0253]

[0254] (1) Preparation method of compound N

[0255] Compound H (509 mg), sodium iodide (300 mg), cuprous iodide (10 mg), 2-pentanol (7 mL), and N,N-dimethylethylenediamine (9 mg) were added sequentially to a flask. The mixture was purged three times with argon gas, and stirred at 140 °C for 12 h under an argon atmosphere. The reaction solution was concentrated under reduced pressure. 20 mL of dichloromethane and 20 mL of water were added to the concentrate, and the mixture was stirred and separated. The organic phase was collected, and the aqueous phase was extracted with 20 mL of dichloromethane. The organic phases were combined and concentrated under reduced pressure to obtain compound N. No purification was required; it was directly added to the next step. ESI-MS: m / z = 557.14 [M+H] + .

[0256] (2) Preparation method of Example 19

[0257] Compound N (100 mg), 1-methyl-4-piperidinol (5 mL), potassium tert-butoxide (100 mg), and copper hydroxide (50 mg) were added to a microwave tube. The microwave power was set to 20 W, the temperature to 150 °C, and the reaction was allowed to proceed for 1.5 h. The reaction solution was diluted with 25 mL of acetonitrile, filtered, and the filtrate was purified by preparative liquid chromatography to obtain Example 19. ESI-MS: m / z = 544.40 [M+H] + .

[0258] Example 20:

[0259]

[0260] (1) Preparation method of compound B-2:

[0261] 1-Bromo-2,5-difluoro-4-nitrobenzene (41.7 mmol), (R)-5-amino-4-methyl-1-pentanol (42.02 mmol), and potassium carbonate (5.80 g) were dissolved in 200 mL of DMF and stirred at room temperature for 4 h until the reaction was complete. The mixture was filtered, and the filter cake was washed with 40 mL of methyl tert-butyl ether. The filtrate was concentrated at 40 °C until no liquid flowed out, yielding the target product B-2. No further purification was required, and it was used directly in the next reaction. ESI-MS: m / z = 335.1 [M+H] + (2) Preparation method of compound C-2:

[0262] Compound B-2 (41.7 mmol) was dissolved in 300 mL of dichloromethane and stirred at room temperature. Triethylamine (6.33 g) was slowly added, and after the addition was complete, the reaction flask was placed in an ice-water bath and stirred for 10 min. Then, methanesulfonyl chloride (5.25 g) was slowly added dropwise. After the addition was complete, the reaction flask was placed in an ice-water bath and stirred for about 2 h until the reaction was complete. The mixture was filtered, and the filter cake was washed with 40 mL of methyl tert-butyl ether. The filtrate was extracted with methyl tert-butyl ether (50 mL x 3) and purified water (50 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated until no liquid flowed out to give compound C-2. No further purification was required, and it was used directly in the next reaction. ESI-MS: m / z = 413.2 [M+H] + .

[0263] (3) Preparation method of compound D-2:

[0264] Compound A (11.8 g), compound C-2 (41.7 mmol), and potassium carbonate (14.4 g) were dissolved in 400 mL of DMF and stirred at 80 °C for 20 min until the reaction was complete. The mixture was filtered, and the filter cake was washed with 40 mL of methyl tert-butyl ether. The filtrate was washed with purified water (50 mL x 3), and the aqueous phase was extracted with methyl tert-butyl ether (50 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated until no liquid eluent was observed. Compound D-2 was purified by column chromatography. ESI-MS: m / z = 564.2 [M+H] + .

[0265] (4) Preparation method of compound E-2:

[0266] Compound D-2 (3.5 g) was dissolved in 300 mL of methanol and stirred at room temperature for 20 min. Raney Ni was slowly added, and the reaction mixture was purged with nitrogen three times, then with hydrogen twice, while stirring under hydrogen atmosphere. The reaction proceeded to completion. The mixture was filtered, and the filter cake was washed with 50 mL of methanol. The filtrate was collected and concentrated at 40 °C until no liquid flowed out, yielding compound E-2. ESI-MS: m / z = 534.2 [M+H] + .

[0267] (5) Preparation method of compound F-2:

[0268] Compound E-2 (1.7 g) and cyanogen bromide (0.7 g) were dissolved in 200 mL of dichloromethane and 200 mL of tert-butanol. The mixture was stirred at room temperature for 18 h. After the reaction was complete, the mixture was washed with purified water (200 mL), retaining the organic phase. The aqueous phase was extracted with 50 mL of dichloromethane. The organic phases were combined and dried over anhydrous sodium sulfate. The mixture was concentrated at 35 °C until no liquid eludes, yielding compound F-2. ESI-MS: m / z = 559.17 [M+H] + .

[0269] (6) Preparation method of compound G-2:

[0270] Compound F-2 (3g) was dissolved in 166mL of tetrahydrofuran solution, and sodium hydroxide (0.9g) was dissolved in 166mL of purified water. The prepared sodium hydroxide aqueous solution was slowly added dropwise to the tetrahydrofuran solution of compound F-2. After the addition was complete, the mixture was stirred at room temperature for 4 hours. After the reaction was complete, the mixture was concentrated at 45°C until no liquid flowed out. 6M hydrochloric acid solution was added dropwise to the concentrate to adjust the pH to 4-5. After the addition was complete, the mixture was stirred at room temperature for 1 hour. The mixture was filtered, and the filter cake was washed with n-hexane to obtain compound G-2. ESI-MS: m / z = 545.16 [M+H] + .

[0271] (7) Preparation method of compound H-2:

[0272] Compound G-2 (1.4 g) was dissolved in 300 mL of dichloromethane, and triethylamine (4.3 g) was slowly added dropwise, followed by TBTU (4.2 g). The mixture was stirred at room temperature for 4 h until the reaction was complete. The reaction solution was washed with 200 mL of purified water, retaining the organic phase. The aqueous phase was extracted with 50 mL of dichloromethane, retaining the organic phase. The organic phase was washed with purified water (20 mL * 3) and dried over anhydrous sodium sulfate. The solution was concentrated until no liquid flowed out, yielding compound H-2. ESI-MS: m / z = 527.24 [M + H] + .

[0273] (8) Preparation method of Example 20:

[0274] Compound H-2 (50 mg), 1-methylpiperidin-4-amine (114 mg), 2-(di-tert-butylphosphine)biphenyl (11.9 mg), tris(dibenzylacetone)dipalladium (18.3 mg), and potassium tert-butoxide (112.5 mg) were sequentially added to tert-amyl alcohol (3 mL). The mixture was purged three times with argon gas, and the reaction was carried out at 115 °C for 2.5 h. The filtrate and concentrate were purified by preparative liquid chromatography to obtain Example 20. ESI-MS: m / z = 561.42 [M+H] + .

[0275] Example 21:

[0276]

[0277] Compound H-2 (100 mg), copper hydroxide (50 mg), and potassium carbonate (50 mg) were dissolved in 1.5 mL of dimethylaminoethanol. The reaction was carried out at 130 °C for 18 h. The reaction was complete. The mixture was extracted with purified water (10 mL x 3) and ethyl acetate (10 mL x 3). The organic phase was dried over anhydrous sodium sulfate. The mixture was concentrated under reduced pressure at 50 °C until no liquid flowed out. The solution was purified by preparative liquid chromatography to obtain Example 21. ESI-MS: m / z = 536.37 [M+H] + .

[0278] Example 22:

[0279]

[0280] (1) Preparation method of compound B-3:

[0281] 1-Bromo-2,5-difluoro-4-nitrobenzene (41.7 mmol), (R)-5-amino-4-methyl-1-pentanol (42.02 mmol), and potassium carbonate (5.80 g) were dissolved in 200 mL of DMF and stirred at room temperature for 4 h until the reaction was complete. The mixture was filtered, and the filter cake was washed with 40 mL of methyl tert-butyl ether. The filtrate was concentrated at 40 °C until no liquid flowed out, yielding the target product B-3. No further purification was required, and it was used directly in the next reaction. ESI-MS: m / z = 351.1 [M+H] + (2) Preparation method of compound C-3:

[0282] Compound B-3 (41.7 mmol) was dissolved in 300 mL of dichloromethane and stirred at room temperature. Triethylamine (6.33 g) was slowly added, and after the addition was complete, the reaction flask was placed in an ice-water bath and stirred for 10 min. Then, methanesulfonyl chloride (5.25 g) was slowly added dropwise. After the addition was complete, the reaction flask was placed in an ice-water bath and stirred for about 2 h until the reaction was complete. The mixture was filtered, and the filter cake was washed with 40 mL of methyl tert-butyl ether. The filtrate was extracted with methyl tert-butyl ether (50 mL x 3) and purified water (50 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated until no liquid flowed out to give compound C-3. No further purification was required, and it was used directly in the next reaction. ESI-MS: m / z = 429.2 [M+H] + .

[0283] (3) Preparation method of compound D-3:

[0284] Compound A (11.8 g), compound C-3 (41.7 mmol), and potassium carbonate (14.4 g) were dissolved in 400 mL of DMF and stirred at 80 °C for approximately 20 min until the reaction was complete. The mixture was filtered, and the filter cake was washed with 40 mL of methyl tert-butyl ether. The filtrate was washed with purified water (50 mL x 3), and the aqueous phase was extracted with methyl tert-butyl ether (50 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated until no liquid eluent was observed. Compound D-3 was purified by column chromatography. ESI-MS: m / z = 580.2 [M+H] + .

[0285] (4) Preparation method of compound E-3:

[0286] Compound D-3 (2.0 g) was dissolved in 300 mL of methanol and stirred at room temperature for 20 min to ensure complete dispersion. Raney Ni was slowly added, and the reaction mixture was purged with nitrogen three times, followed by hydrogen twice. The reaction was stirred under hydrogen atmosphere until complete. The mixture was filtered, and the filter cake was washed with 50 mL of methanol. The filtrate was collected and concentrated at 40 °C until no liquid flowed out, yielding compound E-3. ESI-MS: m / z = 550.13 [M+H] + .

[0287] (5) Preparation method of compound F-3:

[0288] Compound E-3 (1.5 g) and cyanogen bromide (0.6 g) were dissolved in 200 mL of dichloromethane and 200 mL of tert-butanol. The mixture was stirred at room temperature for 18 h. After the reaction was complete, the mixture was washed with purified water (200 mL), retaining the organic phase. The aqueous phase was extracted with 50 mL of dichloromethane. The organic phases were combined and dried over anhydrous sodium sulfate. The mixture was concentrated at 35 °C until no liquid eludes, yielding compound F-3. ESI-MS: m / z = 575.13 [M+H] + .

[0289] (6) Preparation method of compound G-3:

[0290] Compound F-3 (2.5 g) was dissolved in 166 mL of tetrahydrofuran solution, and sodium hydroxide (0.7 g) was dissolved in 166 mL of purified water. The prepared sodium hydroxide aqueous solution was slowly added dropwise to the tetrahydrofuran solution of compound F-3. After the addition was complete, the mixture was stirred at room temperature for 4 h. After the reaction was complete, the mixture was concentrated at 45 °C until no liquid flowed out. 6 M hydrochloric acid solution was added dropwise to the concentrate to adjust the pH to 4-5. After the addition was complete, the mixture was stirred at room temperature for 1 h. The mixture was filtered, and the filter cake was washed with n-hexane to obtain compound G-3. ESI-MS: m / z = 561.18 [M+H] + .

[0291] (7) Preparation method of compound H-3:

[0292] Compound G-3 (1.2 g) was dissolved in 300 mL of dichloromethane, and triethylamine (0.9 g) was slowly added dropwise. Then TBTU (0.9 g) was added, and the mixture was stirred at room temperature for 4 h. After the reaction was complete, the reaction solution was washed with 200 mL of purified water, retaining the organic phase. The aqueous phase was extracted with 50 mL of dichloromethane, retaining the organic phase. The organic phase was washed with purified water (20 mL * 3) and dried over anhydrous sodium sulfate. The solution was concentrated until no liquid flowed out, yielding compound H-3. ESI-MS: m / z = 543.19 [M+H] + .

[0293] (8) Preparation method of Example 22:

[0294] Compound H-3 (50 mg), 1-methylpiperidin-4-amine (114 mg), 2-(di-tert-butylphosphine)biphenyl (11.9 mg), tris(dibenzylacetone)dipalladium (18.3 mg), and potassium tert-butoxide (112.5 mg) were sequentially added to tert-amyl alcohol (3 mL). The mixture was purged three times with argon gas, and the reaction was carried out at 115 °C for 2.5 h. The filtrate and concentrate were purified by preparative liquid chromatography to obtain Example 22. ESI-MS: m / z = 577.39 [M+H] +.

[0295] Example 23:

[0296]

[0297] Compound H-3 (100 mg), copper hydroxide (50 mg), and potassium carbonate (50 mg) were dissolved in 1.5 mL of dimethylaminoethanol. The reaction was carried out at 130 °C for 18 h until complete. The mixture was extracted with purified water (10 mL x 3) and ethyl acetate (10 mL x 3). The organic phase was dried over anhydrous sodium sulfate. The mixture was concentrated under reduced pressure at 50 °C until no liquid eluent was observed. The solution was purified by preparative liquid chromatography to obtain Example 23. ESI-MS: m / z = 552.24 [M+H] + .

[0298] Example 24

[0299]

[0300] (1) Preparation method of compound 2-bromo-4-fluoro-5-nitrotrifluorotoluene

[0301] 4.8 g of 2-bromo-4-fluorotrifluorotoluene was dissolved in 50 mL of concentrated sulfuric acid and cooled to 0–5 °C. Potassium nitrate (2.3 g) was added while maintaining the temperature below 5 °C, and the mixture was stirred at 0–5 °C for 30 min, then at room temperature for 5 h. The reaction mixture was added to 500 mL of cold water, and extracted twice with ethyl acetate. The ethyl acetate extracts were combined and dried over anhydrous sodium sulfate (50 g). The mixture was filtered, and the filtrate was concentrated under reduced pressure to obtain the compound 2-bromo-4-fluoro-5-nitrotrifluorotoluene, which was directly added to the next step.

[0302] (2) Preparation method of compound B-4

[0303] Compound 2-bromo-4-fluoro-5-nitrotrifluorotoluene (3.97 g), potassium carbonate (4.19 g), and DMF (30 mL) were added to a three-necked flask and cooled to 0–5 °C. 2-Methyl-5-aminopentanol (2 g) was dissolved in 30 mL of DMF and added dropwise to the three-necked flask. The mixture was then stirred at room temperature for 2 h. 100 mL of water and 100 mL of ethyl acetate were added to the reaction mixture, and the mixture was stirred and separated, collecting the organic phase. Anhydrous sodium sulfate was added to the organic phase for drying, and the mixture was filtered. The filtrate was concentrated under reduced pressure to obtain compound B-4, which was directly added to the next step. ESI-MS: m / z = 385.2 [M+H] + .

[0304] (3) Preparation method of compound C-4

[0305] Compound B-4 (6.3 g), triethylamine (4.14 g), and tetrahydrofuran (150 mL) were added to a three-necked flask. The mixture was cooled to 0–5 °C, and methanesulfonyl chloride (3.75 g) was added dropwise while maintaining the temperature below 5 °C. After the addition was complete, the mixture was stirred at room temperature for 2 h. The reaction solution was concentrated under reduced pressure, and 100 mL of dichloromethane and 100 mL of water were added to the concentrate. The mixture was stirred and separated, and the organic phase was collected and dried over anhydrous sodium sulfate. The mixture was filtered, and the filtrate was concentrated under reduced pressure to obtain compound C-4, which was directly added to the next step. ESI-MS: m / z = 463.3 [M+H] + .

[0306] (4) Preparation method of compound D-4

[0307] Compound C-4 (7.3 g), compound A (3.44 g), potassium carbonate (4.2 g), and DMF (160 mL) were added to a flask and stirred at 65 °C for 10 h. 250 mL of methyl tert-butyl ether and 250 mL of water were added to the reaction mixture, and the mixture was stirred and separated, collecting the organic phase. The aqueous phase was extracted with 250 mL of methyl tert-butyl ether, and the organic phases were combined. The mixture was then washed successively with 200 mL of water and 200 mL of saturated brine, and finally dried over anhydrous sodium sulfate. The mixture was filtered, and the filtrate was concentrated under reduced pressure to obtain crude compound D-4. Compound D-4 was purified by column chromatography. ESI-MS: m / z = 614.4 [M+H] + .

[0308] (5) Preparation method of compound E-4

[0309] Compound D-4 (3 g), Raney Ni (100 mg), and ethanol (300 mL) were added to a three-necked flask. The mixture was purged with hydrogen three times, stirred at room temperature for 8 hours under a hydrogen atmosphere, filtered, and the filtrate was concentrated under reduced pressure to obtain compound E-4, which was directly added to the next step. ESI-MS: m / z = 584.4 [M+H] + .

[0310] (6) Preparation method of compound F-4

[0311] Compound E-4 (4.3 g), dichloromethane (100 mL), tert-butanol (100 mL), and bromonitrile (1.56 g) were added to a flask and stirred at room temperature for 36 h. 100 mL of dichloromethane and 100 mL of saturated sodium bicarbonate aqueous solution were added to the reaction mixture, and the mixture was stirred and separated. The organic phase was collected and concentrated under pressure to obtain compound F-4, which was directly added to the next step. ESI-MS: m / z = 609.5 [M+H] + .

[0312] (7) Preparation of compound G-4

[0313] Compound F-4 (6.6 g) and tetrahydrofuran (100 mL) were added to a flask and stirred until dissolved. While stirring, a mixed solution of sodium hydroxide (1.7 g) and water (100 mL) was slowly added dropwise. After the addition was complete, the mixture was stirred at room temperature for 4 hours. The reaction solution was concentrated under reduced pressure until no more droplets flowed out. 150 mL of water was added, and the pH was adjusted to approximately 5 by adding 2 mol / L hydrochloric acid solution. Solids were washed out of the solution. The mixture was filtered, and the filter cake was dried under reduced pressure at 50 °C to obtain compound G-4. ESI-MS: m / z = 595.4 [M+H] + .

[0314] (8) Preparation method of compound H-4:

[0315] Compound G-4 (500 mg) was dissolved in 20 mL of acetonitrile, and NMI (241.3 mg) was slowly added dropwise. TCFH (248 mg) was then added, and the mixture was stirred at room temperature for 4 h. After the reaction was complete, the reaction solution was washed with 20 mL of purified water, retaining the organic phase. The aqueous phase was extracted with 50 mL of dichloromethane, retaining the organic phase. The organic phase was washed with purified water (20 mL * 3) and dried over anhydrous sodium sulfate. The solution was concentrated until no liquid flowed out, yielding compound H-4. ESI-MS: m / z = 577.12 [M+H] + .

[0316] (8) Preparation method of Example 24:

[0317] Compound H-4 (400 mg), copper hydroxide (150 mg), and potassium carbonate (250 mg) were dissolved in 1.5 mL of ethylene glycol monomethyl ether. The mixture was placed in a microwave reactor and reacted at 150 °C for 8 h. The reaction was complete. The mixture was extracted with purified water (10 mL x 3) and ethyl acetate (10 mL x 3). The organic phase was dried over anhydrous sodium sulfate. The mixture was concentrated under reduced pressure at 50 °C until no liquid flowed out. The solution was purified by preparative liquid chromatography to obtain Example 24. ESI-MS: m / z = 586.28 [M+H] + .

[0318] Example 25:

[0319]

[0320] Compound H-4 (300 mg), 1-methylpiperidin-4-amine (250 mg), 2-(di-tert-butylphosphine)biphenyl (78 mg), tris(dibenzylacetone)dipalladium (120 mg), and potassium tert-butoxide (350 mg) were sequentially added to tert-amyl alcohol (15 mL). The mixture was purged three times with argon gas, and the reaction was carried out at 115 °C for 2.5 h. The residue was filtered, concentrated, and purified by preparative liquid chromatography to obtain Example 25. ESI-MS: m / z = 611.31 [M+H] + .

[0321] Example 26:

[0322]

[0323] Compound H (200 mg), (S)-3-pyrrolidone (348 mg), 2-(di-tert-butylphosphine)biphenyl (120 mg), tris(dibenzylacetone)dipalladium (184 mg), and potassium tert-butoxide (268 mg) were sequentially added to tert-amyl alcohol (20 mL). The mixture was purged three times with argon gas, and the temperature was raised to 110 °C for 5 h. The residue was filtered, concentrated, and purified by preparative liquid chromatography to obtain Example 26. ESI-MS: m / z = 516.41 [M+H] + .

[0324] 1 H NMR (500MHz, DMSO-d6) δ8.42(s,1H),7.92(s,1H),7.55(d,J=1.2Hz,1H),7.34(d,J=8.6Hz,1H),6.62(d,J=2.2Hz,1H),6.46(dd,J=8.7,2. 1Hz,1H),4.43(tt,J=5.1,2.9Hz,1H),4.36(td,J=9.3,4.5Hz,1H),4.14(dd,J=13.8,3.2Hz,1H),4.02–3.96(m,1H),3.92(dd,J=13.6,10.3 Hz,1H),3.73(s,3H),3.46(dd,J=10.0,5.0Hz,1H),3.42–3.33(m,1H),3.29(td,J=8.5,3.8Hz,1H),3.11(dd,J=9.9,2.4Hz,1H),2.86–2.7 2(m,1H),2.55(s,3H),2.28–2.16(m,1H),2.08(ddt,J=13.2,8.2,4.1Hz,1H),2.02–1.87(m,2H),1.53–1.39(m,1H),0.82(d,J=6.6Hz,3H).

[0325] Example 27:

[0326]

[0327] Compound H (200 mg), (R)-3-pyrrolidone (348 mg), 2-(di-tert-butylphosphine)biphenyl (120 mg), tris(dibenzylacetone)dipalladium (184 mg), and potassium tert-butoxide (268 mg) were sequentially added to tert-amyl alcohol (20 mL). The mixture was purged three times with argon gas, and the reaction was carried out at 110 °C for 4 h. The residue was filtered, concentrated, and purified by preparative liquid chromatography to obtain Example 27. ESI-MS: m / z = 516.45 [M+H]+ .

[0328] 1 H NMR (500MHz, DMSO-d6) δ8.42(s,1H),7.92(s,1H),7.55(s,1H),7.34(d,J=8.6Hz,1H),6.62(d,J=2.1Hz,1H),6.46(dd,J=8.8,2.1Hz ,1H),4.43(tt,J=5.1,2.8Hz,1H),4.36(td,J=9.2,4.5Hz,1H),4.14(dd,J=13.8,3.2Hz,1H),4.02–3.96(m,1H),3.92(dd,J=13.6,1 0.4Hz,1H),3.73(s,3H),3.45(dd,J=10.0,5.0Hz,1H),3.40–3.34(m,2H),3.30(td,J=8.6,3.7Hz,2H),3.12(dd,J=9.9,2.4Hz,1H), 2.87–2.71(m,1H),2.55(s,3H),2.26–2.12(m,1H),2.13–2.03(m,1H),2.02–1.86(m,3H),1.53–1.39(m,1H),0.82(d,J=6.6Hz,3H).

[0329] The compounds of this application can be prepared through the above embodiments. In addition, those skilled in the art can also prepare the following compounds using similar routes, reagents, or methods with reference to the above embodiments:

[0330]

[0331] Experimental Example 1: In vitro kinase activity

[0332] 1.1 Screening for EGFR (WT) inhibitory activity

[0333] Dilute 50 ng / μL of EGFR (WT, Carna) stock solution with kinase buffer (50 mM HEPES, 10 mM MgCl2, 2 mM DTT, 1 mM EGTA, 0.01% Tween 20). Add 6 μL of 1.67× 0.005 ng / μL working solution to each well (final concentration 0.003 ng / μL). Add different compounds dissolved in DMSO to the wells using a nanoparticle pipette to achieve a final concentration gradient of 100 nM to 0.0244 nM, for a total of 7 concentrations. Also include blank control wells (without enzyme) and negative control wells (containing enzyme, with DMSO as solvent), with 2 replicates. After the enzyme reacts with the compound or solvent for 30 min, 5× 25 μM ATP (final concentration 5 μM, Sigma) prepared with kinase buffer and 5× 0.5 μM substrate (final concentration 0.1 μM, Ulight-polyGT, PerkinElmer) are mixed at a 1:1 ratio and added to each well at a concentration of 4 μL. After sealing with a sealing membrane, the plate is incubated at room temperature for 2 h. Then, 5 μL of 4× 40 mM EDTA (final concentration 10 mM) is added to each well, and the plate is incubated at room temperature for 5 min. Next, 5 μL of 4× 8 nM detection reagent (final concentration 2 nM, Eu-anti-phospho-tyrosine antibody, PerkinElmer) is added to each well, and the plate is incubated at room temperature for 1 hour. The plate is then read using a PerkinElmer Envision multi-mode microplate reader (excitation 320 nm, emission 665 nm). The IC50 is calculated using a four-parameter fitting method. 50 .

[0334] 1.2 Screening for EGFR (L858R / T790M) inhibitory activity

[0335] Dilute 50 ng / μL of EGFR (L858R / T790M, Carna) stock solution with kinase buffer (50 mM HEPES, 10 mM MgCl2, 2 mM DTT, 1 mM EGTA, 0.01% Tween 20). Add 6 μL of 1.67× 0.004175 ng / μL working solution to each well (final concentration 0.0025 ng / μL). Add different compounds dissolved in DMSO to the wells using a nanoparticle pipette to achieve a final concentration gradient of 10 nM-0.0024 nM, for a total of 7 concentrations. Also include blank control wells (without enzyme) and negative control wells (containing enzyme, with DMSO as solvent), with 2 replicates. After the enzyme reacts with the compound or solvent for 30 min, 5× 25 μM ATP (final concentration 5 μM, Sigma) prepared with kinase buffer and 5× 0.5 μM substrate (final concentration 0.1 μM, Ultra-poly GT, PerkinElmer) are mixed at a 1:1 ratio and added to each well at a concentration of 4 μL. After sealing with a sealing membrane, the plate is incubated at room temperature for 2 h. Then, 5 μL of 4× 40 mM EDTA (final concentration 10 mM) is added to each well, and the plate is incubated at room temperature for 5 min. Next, 5 μL of 4× 8 nM detection reagent (final concentration 2 nM, Eu-anti-phospho-tyrosine antibody, PerkinElmer) is added to each well, and the plate is incubated at room temperature for 1 hour. The plate is then read using a PerkinElmer Envision multi-mode microplate reader (excitation 320 nm, emission 665 nm). The IC50 is calculated using a four-parameter fitting method. 50 .

[0336] 1.3 Screening for EGFR (d746-750 / T790M) inhibitory activity

[0337] Dilute 50 ng / μL of EGFR (d746-750 / T790M, Carna) stock solution with kinase buffer (50 mM HEPES, 10 mM MgCl2, 2 mM DTT, 1 mM EGTA, 0.01% Tween 20). Add 6 μL of 1.67× 0.005 ng / μL working solution to each well (final concentration 0.003 ng / μL). Add different compounds dissolved in DMSO to the wells using a nanoparticle pipette to achieve a final concentration gradient of 10 nM-0.0024 nM, for a total of 7 concentrations. Also include blank control wells (without enzyme) and negative control wells (containing enzyme, with DMSO as solvent), with 2 replicates. After the enzyme reacts with the compound or solvent for 30 min, 5× 25 μM ATP (final concentration 5 μM, Sigma) prepared with kinase buffer and 5× 0.5 μM substrate (final concentration 0.1 μM, Ultra-poly GT, PerkinElmer) are mixed at a 1:1 ratio and added to each well at a concentration of 4 μL. After sealing with a sealing membrane, the plate is incubated at room temperature for 2 h. Then, 5 μL of 4× 40 mM EDTA (final concentration 10 mM) is added to each well, and the plate is incubated at room temperature for 5 min. Next, 5 μL of 4× 8 nM detection reagent (final concentration 2 nM, Eu-anti-phospho-tyrosine antibody, PerkinElmer) is added to each well, and the plate is incubated at room temperature for 1 hour. The plate is then read using a PerkinElmer Envision multi-mode microplate reader (excitation 320 nm, emission 665 nm). The IC50 is calculated using a four-parameter fitting method. 50 .

[0338] 1.4 EGFR (L858R / T790M / C797S) Inhibitory Activity Screening

[0339] Dilute 50 ng / μL of EGFR (L858R / T790M / C797S, BPS) stock solution with kinase buffer (50 mM HEPES, 10 mM MgCl2, 2 mM DTT, 1 mM EGTA, 0.01% Tween 20). Add 6 μL of 1.67× 0.00167 ng / μL working solution to each well (final concentration 0.001 ng / μL). Add different compounds dissolved in DMSO to the wells using a nanoparticle pipette to achieve a final concentration gradient of 10 nM-0.0024 nM, for a total of 7 concentrations. Also include blank control wells (without enzyme) and negative control wells (containing enzyme, with DMSO as solvent), with 2 replicates. After the enzyme reacts with the compound or solvent for 30 min, 5× 25 μM ATP (final concentration 5 μM, Sigma) prepared with kinase buffer and 5× 0.5 μM substrate (final concentration 0.1 μM, Ultra-poly GT, PerkinElmer) are mixed at a 1:1 ratio and added to each well at a concentration of 4 μL. After sealing with a sealing membrane, the plate is incubated at room temperature for 2 h. Then, 5 μL of 4× 40 mM EDTA (final concentration 10 mM) is added to each well, and the plate is incubated at room temperature for 5 min. Next, 5 μL of 4× 8 nM detection reagent (final concentration 2 nM, Eu-anti-phospho-tyrosine antibody, PerkinElmer) is added to each well, and the plate is incubated at room temperature for 1 hour. The plate is then read using a PerkinElmer Envision multi-mode microplate reader (excitation 320 nm, emission 665 nm). The IC50 is calculated using a four-parameter fitting method. 50 .

[0340] Screening for 1.5 EGFR (d746-750 / T790M / C797S) inhibitory activity

[0341] Dilute 50 ng / μL of EGFR (d746-750 / T790M / C797S, BPS) stock solution with kinase buffer (50 mM HEPES, 10 mM MgCl2, 2 mM DTT, 1 mM EGTA, 0.01% Tween 20). Add 6 μL of 1.67× 0.05 ng / μL working solution to each well (final concentration 0.03 ng / μL). Add different compounds dissolved in DMSO to the wells using a nanoparticle pipette to achieve a final concentration gradient of 10 nM-0.0024 nM, for a total of 7 concentrations. Also include blank control wells (without enzyme) and negative control wells (containing enzyme, with DMSO as solvent), with 2 replicates. After the enzyme reacts with the compound or solvent for 30 min, 5× 25 μM ATP (final concentration 5 μM, Sigma) prepared with kinase buffer and 5× 0.5 μM substrate (final concentration 0.1 μM, Ultra-poly GT, PerkinElmer) are mixed at a 1:1 ratio and added to each well at a concentration of 4 μL. After sealing with a sealing membrane, the plate is incubated at room temperature for 2 h. Then, 5 μL of 4× 40 mM EDTA (final concentration 10 mM) is added to each well, and the plate is incubated at room temperature for 5 min. Next, 5 μL of 4× 8 nM detection reagent (final concentration 2 nM, Eu-anti-phospho-tyrosine antibody, PerkinElmer) is added to each well, and the plate is incubated at room temperature for 1 hour. The plate is then read using a PerkinElmer Envision multi-mode microplate reader (excitation 320 nm, emission 665 nm). The IC50 is calculated using a four-parameter fitting method. 50 The results are shown in Table 1:

[0342] Table 1 Results of in vitro kinase activity

[0343]

[0344]

[0345] Experimental Example 2: In vitro evaluation of liver microsite stability

[0346] The final 300 μL incubation system contained 30 μL liver microsomes (protein concentration: 0.15 mg / mL), 30 μL NADPH + MgCl2, 3 μL test compound (prepared with acetonitrile), and 237 μL PBS buffer (pH 7.4). The proportion of organic solvent (acetonitrile) was 1%. Two aliquots were prepared for each species, 0.3 mL each. For each tube, a 270 μL mixture of substrate and enzyme was prepared, and the mixture was pre-incubated with NADPH at 37°C for 5 min. Then, 30 μL of NADPH + MgCl2 was added and mixed. At 0, 15, 30, and 60 min, 50 μL of the mixture was collected and the reaction was terminated with 300 μL of ice-cold acetonitrile containing the internal standard.

[0347] 50 μL of sample was incubated at room temperature, and 300 μL of acetonitrile containing the internal standard (diazepam 20 ng / mL) was added to precipitate the sample. The mixture was vortexed for 5 min and then centrifuged (12000 rpm, 4℃) for 10 min. 75 μL of the supernatant was collected, diluted with 75 μL of ultrapure water, and injected into the sample at 0.5 μL for analysis. Elimination parameters of the relevant compounds in human and mouse liver microsomes are shown in Table 2.

[0348] Table 2. Results of in vitro liver microsite stability evaluation

[0349]

[0350] Example 3: In vivo pharmacokinetic evaluation

[0351] ICR mice, weighing 18–20 g, were acclimatized for 3–5 days and then randomly divided into groups of 9 mice each. Each group was administered the corresponding compound by gavage at a dose of 10 mg / kg.

[0352] The test animals (ICR mice) were fasted for 12 hours before administration and given food 4 hours after administration. They had free access to water before, during, and after the experiment.

[0353] Following oral administration, approximately 0.1 mL of blood was collected from the orbital cavity at 0 min, 15 min, 30 min, 1 h, 2 h, 4 h, 6 h, 8 h, 10 h, and 24 h. After EDTA-K2 anticoagulation, plasma was separated by centrifugation at 4000 rpm for 10 min at 4 °C within 30 min. All collected plasma was immediately stored at -20 °C for analysis. 20 μL of the plasma sample and standard curve were added to 300 μL of acetonitrile solution containing the internal standard (diazepam 20 mg / mL), vortexed for 5 min, centrifuged at 12000 rpm for 10 min, and 70 μL of the supernatant was collected, diluted with 70 μL of ultrapure water, mixed, and 2 μL was used for LC / MS / MS analysis. The chromatogram was recorded. The in vivo pharmacokinetic parameters of the relevant compounds are shown in Table 3.

[0354] Experiments show that the compound of this application has good pharmacokinetic properties in vivo and a high oral exposure.

[0355] Table 3 Pharmacokinetic parameters

[0356] PK parameters Example 27: IG 10 mg / kg Tmax(h) 1.00 Cmax(ng / mL) 262 AUC(0-t)(ng*h / mL) 1853 AUC(0-∞)(ng*h / mL) 1888 t1 / 2(h) 4.13 MRT(0-t)(h) 5.81 .

Claims

1. The following compounds or their pharmaceutically acceptable salts, stereoisomers, and tautomers: or .

2. A pharmaceutical composition comprising the compound of claim 1 or a pharmaceutically acceptable salt, stereoisomer, or tautomer thereof.

3. Use of the compound of claim 1 or a pharmaceutically acceptable salt, stereoisomer, tautomer, or pharmaceutical composition of claim 2 in the preparation of a medicament for the prevention or treatment of EGFR-mediated diseases.

4. The use according to claim 3, wherein the EGFR-mediated disease is selected from EGFR mutation-mediated diseases.

5. The use as described in claim 4, wherein the mutant is selected from one, two, three, or four of L858R, T790M, d19, and C797S.

6. The use as described in claim 4, wherein the mutant is selected from two mutants, L858R and T790M.

7. The use as described in claim 4, wherein the mutant is selected from two mutants, d19 and T790M.

8. The use as claimed in claim 4, wherein the mutant comprises the C797S mutant.

9. The use as described in claim 4, wherein the mutant is selected from three mutants: L858R, T790M, and C797S.

10. The use as described in claim 4, wherein the mutant is selected from three mutants: d19, T790M, and C797S.

11. The use according to claim 3, wherein the EGFR-mediated disease is selected from cancer.

12. The use as described in claim 3, wherein the EGFR-mediated disease is selected from lung cancer.

13. The use as described in claim 3, wherein the EGFR-mediated disease is selected from non-small cell lung cancer.

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