Ulks1 inhibitors, methods of making and uses thereof

Abstract

The present application relates to a new ULK1 inhibitor represented by formula (1), a pharmaceutical composition and kit comprising the same, a preparation method thereof and use thereof in the preparation of a drug for a ULK1-mediated disease.

Description

ULK1 inhibitors and their preparation methods and uses

[0001] Citation of Related Applications

[0002] This application claims priority to Chinese patent application No. 202311187688.4 filed on September 13, 2023, the contents of which are incorporated by reference into this application in their entirety and for all purposes. Technical Field

[0003] The present invention relates to a novel ULK1 inhibitor, a pharmaceutical composition and a pharmaceutical kit containing the same, a preparation method thereof and use thereof in preparing drugs for ULK1-mediated diseases. Background Art

[0004] Autophagy is a central cellular mechanism for the elimination of damaged proteins, protein complexes, and organelles. This conserved process, in addition to being required for proper cellular and tissue homeostasis during embryonic development and in defense against pathogens, plays a crucial role in the cellular response to nutrient deprivation and other stresses. Defects in the autophagic pathway have been implicated in several human pathologies, including infectious diseases, neurodegenerative disorders, and cancer. Despite these highly conserved, fundamental cellular functions, the molecular and biochemical details of how different cargoes initiate autophagy, as well as the coordination of steps from autophagosome priming to eventual fusion with lysosomes, remain poorly understood.

[0005] Provided herein are inhibitors of unc51-like autophagy-activating kinase (ULK) proteins. In many cases, ULK1 and ULK2 are important proteins that regulate autophagy in mammalian cells. In some cases, ULK1 and ULK2 are activated by several upstream signals under conditions of nutrient deprivation, subsequently initiating autophagy. The requirement for ULK1 and ULK2 in the initiation of autophagy has been studied in the context of nutrient deprivation. Although ULK1 appears to be the most essential for autophagy, in some cases, ULK1 and ULK2 show high functional redundancy. The kinase domains of ULK1 and ULK2 share 78% sequence homology, indicating that in some cases, ULK2 may compensate for the loss of ULK1. Therefore, only when both ULK1 and ULK2 are inhibited can nutrient-dependent autophagy be eliminated. However, in some cases, ULK1 inhibition alone is sufficient to provide therapeutic benefits, such as normalizing autophagy in any of the methods provided herein, or other beneficial results. While inhibition of ULK1 and ULK2 produces therapeutic benefits, such as tumor shrinkage, tumor cell death, or reduced tumor growth rate, there remains a need to provide new ULK1 inhibitors for the treatment of ULK1-mediated diseases.

[0006] SUMMARY OF THE INVENTION

[0007] In one aspect, the present invention provides a pyrimidine compound that has a specific inhibitory effect on ULK1, thereby having a good therapeutic effect on ULK1-mediated diseases. The compound is a compound of Formula 1 or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, hydrate, N-oxide, isotope-labeled compound, metabolite, ester, or prodrug thereof:

[0008] Wherein, X1, X2, and X3 are independently CH or N;

[0009] Ring A is selected from 3-10 membered heterocyclic group, C3-C 10 Cycloalkyl, C6-C 10 Aryl and 5-10 membered heteroaryl;

[0010] Ring B is selected from 3-10 membered heterocyclic group, C3-C 10 Cycloalkyl, C6-C 10 Aryl and 5-10 membered heteroaryl;

[0011] R1 is independently selected from H, halogen, cyano, nitro, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, C3-C 10 Cycloalkyl, 3-10 membered heterocyclic group, C6-C 10 Aryl and 5-10 membered heteroaryl;

[0012] R2 is selected from H, cyano, halogen, C1-C6 alkyl, C1-C6 hydroxyalkyl, C1-C6 aminoalkyl, C1-C6 haloalkyl, C1-C6 heteroalkyl, C2-C6 alkenyl, C2-C6 alkynyl and C3-C 10 Cycloalkyl;

[0013] R3 and R4 are each independently selected from H, oxo, halogen, cyano, nitro, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C 10 Cycloalkyl, 3-10 membered heterocyclic group, C6-C 10 aryl, 5-10 membered heteroaryl, R5O-, R6C(O)-, -NR7R8 and -NHCONR7R8, wherein the C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C 10 Cycloalkyl, 3-10 membered heterocyclic group, C6-C 10 Aryl or 5-10 membered heteroaryl are each optionally substituted with one or more R9;

[0014] R5 and R6 are each independently selected from H, C1-C6 alkyl, C3-C 10 Cycloalkyl, C6-C 10 Aryl and 5-10 membered heteroaryl;

[0015] R7 and R8 are each independently selected from H, C1-C6 alkyl, C3-C 10 Cycloalkyl, 3-10 membered heterocyclic and C6-C 10 Aryl; or R7, R8 and the N atom to which they are attached together form a 5-6 membered heterocyclic ring or a 5-6 membered heteroaromatic ring;

[0016] R9 are each independently selected from halogen, cyano, nitro, amino, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C 10 Cycloalkyl, 3-10 membered heterocyclic group, C6-C 10 Aryl and 5-10 membered heteroaryl;

[0017] R is selected from -NR7R8, R5O-, R5S-, halogen, C1-C6 alkyl, 3-12 membered heterocyclyl, 5-10 membered heteroaryl and -LR 10 wherein the 3-12 membered heterocyclyl or 5-10 membered heteroaryl are each optionally substituted by one or more R 11 replace;

[0018] L is -(L1) m -(L2) n -(L3) p -;

[0019] L1 is selected from -O-, -N(R 12 )-、-C(R 12 )(R 13 )-O-、-N(R 12 )CO- and -S-;

[0020] L2 is selected from C1-C6 alkylene, C1-C6 heteroalkylene, C3-C 10 Cycloalkylene and C3-C 10 heterocyclylene;

[0021] L3 is selected from -N(R 12 )-、-N(R 12 )C(O)- and -C(O)-;

[0022] m, n and p are each independently selected from 0, 1 and 2;

[0023] R 10 Selected from C3-C 10 Cycloalkyl, C6-C 10 Aryl, 5-10 membered heteroaryl and 3-10 membered heterocyclic group, the C3-C 10 Cycloalkyl, C6-C 10 Aryl, 5-10 membered heteroaryl or 3-10 membered heterocyclyl are each optionally substituted by one or more R14 replace;

[0024] R 11 Each is independently selected from C1-C6 alkyl, oxo, 5-10 membered heteroaryl, 3-10 membered heterocyclyl, R6C(O)-, R6C(O)N(R 12 )- and -NR7R8, wherein the C1-C6 alkyl, 5-10 membered heteroaryl or 3-10 membered heterocyclyl are each optionally substituted by one or more R 15 replace;

[0025] R 12 、R 13 Each independently selected from H, C1-C6 alkyl and C3-C 10 Cycloalkyl;

[0026] R 14 are each independently selected from H, halogen, cyano, oxo, C1-C6 alkyl, C1-C6 alkoxy, -CONR7R8 and R6C(O)-; and

[0027] R 15 Each independently selected from H, halogen, cyano, oxo, C1-C6 alkyl, C3-C 10 Cycloalkyl and 3-10 membered heterocyclic groups, wherein the C1-C6 alkyl, C3-C 10 The cycloalkyl or 3-10 membered heterocyclyl are each optionally substituted with one or more substituents selected from C1-C6 alkyl and oxo; and

[0028] x, y, z are each independently selected from 1, 2 and 3.

[0029] In another aspect, the present invention provides a pharmaceutical composition comprising an effective amount of a compound of the present invention or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, hydrate, N-oxide, isotopically labeled compound, metabolite, ester, or prodrug thereof, and one or more pharmaceutically acceptable carriers.

[0030] In another aspect, the present invention provides a compound of the present invention or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, hydrate, N-oxide, isotopically labeled compound, metabolite, ester, or prodrug thereof, or a pharmaceutical composition of the present invention for the preparation of a medicament for treating a ULK1-mediated disease in a subject in need thereof.

[0031] In another aspect, the present invention provides a method for treating a ULK1-mediated disease in a subject in need thereof, comprising administering to the subject a compound according to the present invention or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, hydrate, N-oxide, isotopically labeled compound, metabolite, ester, or prodrug thereof, or a pharmaceutical composition of the present invention.

[0032] In another aspect, the present invention provides a compound of the present invention or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, hydrate, N-oxide, isotopically labeled compound, metabolite, ester, or prodrug thereof, or a pharmaceutical composition of the present invention, for use in treating a ULK1-mediated disease in a subject in need thereof.

[0033] In another aspect, the present invention provides methods of preparing the compounds of the present invention.

[0034] Detailed Description of the Invention

[0035] definition

[0036] Unless otherwise defined below, all technical and scientific terms used herein are intended to have the same meaning as those commonly understood by those skilled in the art. References to technology used herein are intended to refer to technology commonly understood in the art, including variations of technology or substitutions of equivalent technology that would be apparent to those skilled in the art. While it is believed that the following terms are well understood by those skilled in the art, the following definitions are set forth to better explain the present invention.

[0037] As used herein, the terms "comprises," "comprising," "having," "containing," or "involving," and variations thereof herein, are inclusive or open-ended and do not exclude additional unrecited elements or method steps.

[0038] As used herein, the term "alkyl" is defined as a straight-chain or branched saturated aliphatic hydrocarbon group. For example, as used herein, the term "C 1-6 "Alkyl" refers to a straight or branched chain group having 1 to 6 carbon atoms (e.g., methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl or n-hexyl).

[0039] As used herein, the term "alkylene" refers to a straight-chain or branched divalent alkyl group.

[0040] As used herein, the term "alkenyl" refers to a straight-chain or branched aliphatic hydrocarbon group having one or more unsaturated double bonds. For example, as used herein, the term "C2-C6 alkenyl" refers to an alkenyl group having 2 to 6 carbon atoms, such as ethenyl, 1-propenyl, 2-propenyl, 2-methyl-1-propenyl, 2-methyl-2-propenyl, 2-buten-1-yl, 3-buten-1-yl, 2-penten-1-yl, 3-penten-1-yl, 4-penten-1-yl, 5-hexen-1-yl, 4-hexen-1-yl, 3-hexen-1-yl, 2-hexen-1-yl, 3-methyl-2-buten-1-yl, 3-methyl-3-penten-1-yl, 3-methyl-2-penten-1-yl, 4-methyl-3-penten-1-yl, 4-methyl-2-penten-1-yl, and 2-methyl-2-penten-1-yl, etc. Preferably, the alkenyl group has one double bond.

[0041] As used herein, the term "alkynyl" refers to a straight or branched aliphatic hydrocarbon group with one or more unsaturated triple bonds. For example, as used herein, the term "C2-C6 alkynyl" refers to an alkenyl group with 2 to 6 carbon atoms, such as ethynyl, 1-propyn-1-yl, 2-propyn-1-yl, 2-butyn-1-yl, 3-butyn-1-yl, 2-pentyn-1-yl, 3-pentyn-1-yl, 4-pentyn-1-yl, 5-hexyn-1-yl, 4-hexyn-1-yl, 3-hexyn-1-yl and 2-hexyn-1-yl. Preferably, it has one triple bond.

[0042] As used herein, the term "alkoxy" refers to a group having an oxygen atom inserted into an alkyl group (as defined above) at any reasonable position, such as C 1-8 Alkoxy, C 1-6 Alkoxy, C 1-4 Alkoxy or C 1-3 Alkoxy. C 1-6 Representative examples of alkoxy include, but are not limited to, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, pentyloxy, hexyloxy, and the like.

[0043] As used herein, the term "halo" or "halogen" group is defined to include fluorine, chlorine, bromine, or iodine.

[0044] As used herein, the term "haloalkyl" refers to an alkyl group substituted by one or more (such as 1 to 3) the same or different halogen atoms. For example, the term "C 1-6 "Haloalkyl" refers to a haloalkyl group having 1 to 6 carbon atoms, for example, -CF3, -C2F5, -CHF2, -CH2F, -CH2CF3, -CH2Cl or -CH2CH2CF3, etc.

[0045] As used herein, the term "hydroxyalkyl" refers to an alkyl group substituted with one or more (such as 1 to 3) hydroxy groups. For example, the term "C 1-6 The term "hydroxyalkyl" refers to a hydroxyalkyl group having 1 to 6 carbon atoms, for example, -CH2OH, -CH2CH2OH, etc.

[0046] As used herein, the term "aminoalkyl" refers to an alkyl group substituted with one or more (such as 1 to 3) amino groups. For example, the term "C 1-6 "Aminoalkyl" refers to an aminoalkyl group having 1 to 6 carbon atoms, for example, -CH2NH2, -CH2CH2NH2, etc.

[0047] As used herein, the term "heteroalkyl" refers to a group consisting of one or more groups independently selected from N, O, or S(O). q (where q is 0, 1 or 2) is a straight or branched alkyl group interrupted by a heteroatom. For example, the term "C 1-6 "Heteroalkyl" refers to a heteroalkyl group having 1 to 6 carbon atoms.

[0048] As used herein, the term "heteroalkylene" refers to a group consisting of one or more groups independently selected from N, O, or S(O). q (wherein q is 0, 1 or 2) is a straight or branched chain alkylene group interrupted by a heteroatom. For example, the term "C 1-6 "Heteroalkylene" refers to a heteroalkylene group having 1 to 6 carbon atoms.

[0049] As used herein, the term "cycloalkyl" refers to a saturated or partially unsaturated non-aromatic monocyclic or polycyclic (such as bicyclic) hydrocarbon ring (e.g., monocyclic, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, or bicyclic, including spirocyclic, fused (and fused) or bridged (bridged) systems, such as bicyclo[1.1.1]pentyl, bicyclo[2.2.1]heptyl, bicyclo[3.2.1]octyl or bicyclo[5.2.0]nonyl, decahydronaphthyl, etc.), which is optionally substituted with one or more (such as 1 to 3) suitable substituents. The cycloalkyl has 3 to 15, for example 3 to 10 carbon atoms. For example, the term "C 3-10 "Cycloalkyl" refers to a saturated or partially unsaturated non-aromatic monocyclic or polycyclic (such as bicyclic) hydrocarbon ring having 3 to 10 ring carbon atoms (for example, cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl).

[0050] As used herein, the term "heterocyclyl" or "heterocycle" refers to a saturated or partially unsaturated monocyclic or polycyclic (including spirocyclic, fused (paracyclic) or bridged (bridged) ring) group, for example, having 2, 3, 4, 5, 6, 7, 8 or 9 carbon atoms and one or more (e.g., 1, 2, 3 or 4) independently selected from N, O or S (O) in the ring. q(wherein q is 0, 1 or 2) heteroatoms, such as 3-12 membered heterocyclyl, 3-10 membered heterocyclyl, 3-6 membered heterocyclyl, 5-6 membered heterocyclyl, etc. Representative examples of heterocyclyl include, but are not limited to, oxiranyl, aziridinyl, azetidinyl, oxetanyl, tetrahydrofuranyl, pyrrolidinyl, pyrrolidonyl, imidazolidinyl, pyrazolidinyl, tetrahydropyranyl, piperidinyl, morpholinyl, dithianyl, thiomorpholinyl, piperazinyl, trithianyl, etc. As used herein, the term "nitrogen-containing heterocycle" refers to a heterocycle having at least one nitrogen atom in the ring and optionally other heteroatoms, including monocyclic or polycyclic (including spirocyclic, fused (parallel ring) or bridged (bridged ring) systems) heterocycles.

[0051] As used herein, the term "aryl" or "aromatic ring" refers to an all-carbon monocyclic or fused-ring polycyclic aromatic group having a conjugated π electron system. For example, the term "C 6-10 Aryl" or "C 6-10 "Aromatic ring" refers to an aromatic group containing 6 to 10 carbon atoms, such as phenyl (ring) or naphthyl (ring).

[0052] As used herein, the term "heteroaryl" or "heteroaromatic ring" refers to a monocyclic, bicyclic or tricyclic aromatic ring system containing at least one heteroatom selected from N, O and S, for example having 5, 6, 8, 9, 10, 11, 12, 13 or 14 ring atoms, in particular containing 1 or 2 or 3 or 4 or 5 or 6 or 9 or 10 carbon atoms, and, in each case, may be benzo-fused. For example, examples of heteroaryl or heteroaromatic rings include, but are not limited to, thienyl (ring), furanyl (ring), pyrrolyl (ring), oxazolyl (ring), thiazolyl (ring), imidazolyl (ring), pyrazolyl (ring), isoxazolyl (ring), isothiazolyl (ring), oxadiazolyl (ring), triazolyl (ring), thiadiazolyl (ring), and the like, and benzo derivatives thereof; or pyridinyl (ring), pyridazinyl (ring), pyrimidinyl (ring), pyrazinyl (ring), triazinyl (ring), and the like, and benzo derivatives thereof.

[0053] The term "substituted" means that one or more (e.g., 1, 2, 3, or 4) hydrogen atoms on the designated atom are replaced with a group selected from the indicated group, provided that the designated atom's normal valence in the current context is not exceeded and that the substitution results in a stable compound. Combinations of substituents and / or variables are permissible only if such combinations result in stable compounds.

[0054] If a substituent is described as being "optionally substituted with," the substituent may be (1) unsubstituted or (2) substituted. If a carbon of a substituent is described as being optionally substituted with one or more of the substituents listed, one or more hydrogens on the carbon (to the extent of any hydrogens present) may be replaced, individually and / or collectively, with independently selected substituents or unsubstituted. If a nitrogen of a substituent is described as being optionally substituted with one or more of the substituents listed, one or more hydrogens on the nitrogen (to the extent of any hydrogens present) may each be replaced with an independently selected substituent or unsubstituted.

[0055] If a substituent is described as being "independently selected" from a group of groups, each substituent is selected independently of the other. Thus, each substituent may be the same as or different from another (other) substituent.

[0056] As used herein, the term "one or more" means 1 or more than 1, such as 2, 3, 4, 5, 6, 7, 8, 9 or 10, where reasonable.

[0057] Unless otherwise indicated, as used herein, the point of attachment of a substituent may be from any suitable position of the substituent.

[0058] When a bond to a substituent is shown to pass through a bond connecting two atoms in a ring, then such substituent may be bonded to any ring atom in the substitutable ring.

[0059] The present invention also includes all pharmaceutically acceptable isotopically labeled compounds which are identical to the compounds of the present invention except that one or more atoms are replaced by an atom having the same atomic number but an atomic mass or mass number different from the atomic mass or mass number prevalent in nature. Examples of suitable isotopes for inclusion in the compounds of the present invention include, but are not limited to, isotopes of hydrogen (e.g., 2 H. 3 H, deuterium D, tritium T); carbon isotopes (such as 11 C. 13 C and 14 C); isotopes of chlorine (e.g. 37 Cl); isotopes of fluorine (e.g. 18 F); isotopes of iodine (such as 123 I and 125 I); isotopes of nitrogen (e.g. 13 N and 15 N); oxygen isotopes (e.g. 15 O. 17 O and 18 O); isotopes of phosphorus (such as 32 P); and sulfur isotopes (e.g. 35S). Certain isotopically labeled compounds of the invention (e.g., those incorporating radioactive isotopes) are useful in drug and / or substrate tissue distribution studies (e.g., assays). The radioactive isotope tritium (i.e., 3 H) and carbon-14 (i.e. 14 C) are particularly useful for this purpose because they are easy to incorporate and easy to detect. 11 C. 18 F. 15 O and 13 N) substitution can be used to examine substrate receptor occupancy in positron emission tomography (PET) studies. Isotopically labeled compounds of the present invention can be prepared by methods analogous to those described in the accompanying schemes and / or examples and preparations by using appropriate isotopically labeled reagents instead of the non-labeled reagents previously employed. Pharmaceutically acceptable solvates of the present invention include those in which the crystallization solvent is isotopically substituted, for example, D2O, acetone-d6 or DMSO-d6.

[0060] The term "stereoisomer" refers to an isomer formed due to at least one asymmetric center. In compounds with one or more (e.g., 1, 2, 3, or 4) asymmetric centers, racemic mixtures, single enantiomers, diastereomeric mixtures, and individual diastereomers may occur. Specific individual molecules may also exist as geometric isomers (cis / trans).

[0061] Similarly, the compound of the present invention can exist as a mixture (commonly referred to as tautomer) of two or more structurally different forms in rapid equilibrium. Representative examples of tautomers include keto-enol tautomers, phenol-ketone tautomers, nitroso-oxime tautomers, imines-enamine tautomers, etc. It is understood that the scope of the present application encompasses all such isomers or mixtures thereof in any proportion (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%).

[0062] The present invention encompasses all possible crystalline forms or polymorphs of the compounds of the present invention, which may be single polymorphs or mixtures of more than one polymorph in any ratio.

[0063] It should also be understood that certain compounds of the present invention may be used therapeutically in free form or, where appropriate, in the form of pharmaceutically acceptable derivatives thereof. In the present invention, pharmaceutically acceptable derivatives include, but are not limited to, pharmaceutically acceptable salts, solvates, metabolites, or prodrugs that, upon administration to a patient in need thereof, are capable of directly or indirectly providing a compound of the present invention or a metabolite or residue thereof. Therefore, when reference is made herein to a "compound of the present invention," such various derivative forms of the compound are also intended to be encompassed.

[0064] Pharmaceutically acceptable salts of the compounds of the present invention include acid addition salts and base addition salts thereof. Suitable acid addition salts are formed from acids that form pharmaceutically acceptable salts. Suitable base addition salts are formed from bases that form pharmaceutically acceptable salts. For a review of suitable salts, see Stahl and Wermuth, "Handbook of Pharmaceutical Salts: Properties, Selection, and Use" (Wiley-VCH, 2002). Methods for preparing pharmaceutically acceptable salts of the compounds of the present invention are known to those skilled in the art.

[0065] The compounds of the present invention may exist in the form of solvates (preferably hydrates), wherein the compounds of the present invention contain a polar solvent as a structural element of the crystal lattice of the compound, in particular water, methanol or ethanol. The amount of polar solvent, in particular water, may be present in a stoichiometric or non-stoichiometric ratio.

[0066] Those skilled in the art will appreciate that not all nitrogen-containing heterocycles are capable of forming N-oxides, as nitrogen requires an available lone pair of electrons to oxidize to an oxide; those skilled in the art will recognize nitrogen-containing heterocycles that are capable of forming N-oxides. Those skilled in the art will also recognize that tertiary amines are capable of forming N-oxides. Synthetic methods for preparing N-oxides of heterocycles and tertiary amines are well known to those skilled in the art and include oxidation of heterocycles and tertiary amines with peroxyacids such as peracetic acid and meta-chloroperbenzoic acid (MCPBA), hydrogen peroxide, alkyl hydroperoxides such as tert-butyl hydroperoxide, sodium perborate, and dioxirane such as dimethyldioxirane. These methods for preparing N-oxides have been extensively described and reviewed in the literature, see for example: TL Gilchrist, Comprehensive Organic Synthesis, vol. 7, pp 748-750; AR Katritzky and AJ Boulton, Eds., Academic Press; and GWH Cheeseman and ESGWerstiuk, Advances in Heterocyclic Chemistry, vol. 22, pp 390-392, AR Katritzky and AJ Boulton, Eds., Academic Press.

[0067] Also included within the scope of the present invention are metabolites of the compounds of the invention, i.e., substances formed in vivo upon administration of the compounds of the invention. Such products may be produced, for example, by oxidation, reduction, hydrolysis, amidation, deamidation, esterification, enzymatic hydrolysis, etc. of the administered compound. Thus, the present invention includes metabolites of the compounds of the invention, including compounds produced by contacting a compound of the invention with a mammal for a period of time sufficient to produce a metabolic product thereof.

[0068] The present invention further includes within its scope prodrugs of the compounds of the present invention, which are certain derivatives of the compounds of the present invention that may themselves have little or no pharmacological activity, and when administered to the body or thereon, can be converted into the compounds of the present invention having the desired activity by, for example, hydrolytic cleavage. Typically, such prodrugs will be functional group derivatives of the compounds that are readily converted into the desired therapeutically active compounds in vivo. Further information on the use of prodrugs can be found in "Pro-drugs as Novel Delivery Systems," Volume 14, ACS Symposium Series (T. Higuchi and V. Stella) and "Bioreversible Carriers in Drug Design," Pergamon Press, 1987 (E.B. Roche, ed., American Pharmaceutical Association). The prodrugs of the present invention can be prepared, for example, by replacing appropriate functional groups present in the compounds of the present invention with certain moieties known to those skilled in the art as "pro-moieties" (e.g., as described in "Design of Prodrugs," H. Bundgaard (Elsevier, 1985)).

[0069] The present invention also encompasses compounds of the present invention that contain protecting groups. During any process for preparing the compounds of the present invention, it may be necessary and / or desirable to protect sensitive or reactive groups on any of the molecules involved, thereby forming a chemically protected form of the compounds of the present invention. This can be achieved using conventional protecting groups, for example, those described in Protective Groups in Organic Chemistry, ed. JFW McOmie, Plenum Press, 1973; and TW Greene & P.GM Wuts, Protective Groups in Organic Synthesis, John Wiley & Sons, 1991, which references are incorporated herein by reference. Protecting groups can be removed at an appropriate subsequent stage using methods known in the art.

[0070] The term "about" means within ±10%, preferably within ±5%, and more preferably within ±2% of the stated numerical value.

[0071] Compound

[0072] One object of the present invention is to provide a compound of Formula 1 or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, hydrate, N-oxide, isotope-labeled compound, metabolite, ester, or prodrug thereof:

[0073] Wherein, X1, X2, and X3 are independently CH or N;

[0074] Ring A is selected from 3-10 membered heterocyclic group, C3-C 10 Cycloalkyl, C6-C 10 Aryl and 5-10 membered heteroaryl;

[0075] Ring B is selected from 3-10 membered heterocyclic group, C3-C 10 Cycloalkyl, C6-C 10 Aryl and 5-10 membered heteroaryl;

[0076] R1 is independently selected from H, halogen, cyano, nitro, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, C3-C 10 Cycloalkyl, 3-10 membered heterocyclic group, C6-C 10 Aryl and 5-10 membered heteroaryl;

[0077] R2 is selected from H, cyano, halogen, C1-C6 alkyl, C1-C6 hydroxyalkyl, C1-C6 aminoalkyl, C1-C6 haloalkyl, C1-C6 heteroalkyl, C2-C6 alkenyl, C2-C6 alkynyl and C3-C 10 Cycloalkyl;

[0078] R3 and R4 are each independently selected from H, oxo, halogen, cyano, nitro, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C 10 Cycloalkyl, 3-10 membered heterocyclic group, C6-C 10 aryl, 5-10 membered heteroaryl, R5O-, R6C(O)-, -NR7R8 and -NHCONR7R8, wherein the C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C 10 Cycloalkyl, 3-10 membered heterocyclic group, C6-C 10 Aryl or 5-10 membered heteroaryl are each optionally substituted with one or more R9;

[0079] R5 and R6 are each independently selected from H, C1-C6 alkyl, C3-C 10 Cycloalkyl, C6-C 10 Aryl and 5-10 membered heteroaryl;

[0080] R7 and R8 are each independently selected from H, C1-C6 alkyl, C3-C10 Cycloalkyl, 3-10 membered heterocyclic and C6-C 10 Aryl; or R7, R8 and the N atom to which they are attached together form a 5-6 membered heterocyclic ring or a 5-6 membered heteroaromatic ring;

[0081] R9 are each independently selected from halogen, cyano, nitro, amino, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C 10 Cycloalkyl, 3-10 membered heterocyclic group, C6-C 10 Aryl and 5-10 membered heteroaryl;

[0082] R is selected from -NR7R8, R5O-, R5S-, halogen, C1-C6 alkyl, 3-12 membered heterocyclyl, 5-10 membered heteroaryl and -LR 10 wherein the 3-12 membered heterocyclyl or 5-10 membered heteroaryl are each optionally substituted by one or more R 11 replace;

[0083] L is -(L1) m -(L2) n -(L3) p -;

[0084] L1 is selected from -O-, -N(R 12 )-、-C(R 12 )(R 13 )-O-、-N(R 12 )CO- and -S-;

[0085] L2 is selected from C1-C6 alkylene, C1-C6 heteroalkylene, C3-C 10 Cycloalkylene and 3-10 membered heterocyclylene;

[0086] L3 is selected from -N(R 12 )-、-N(R 12 )C(O)- and -C(O)-;

[0087] m, n and p are each independently selected from 0, 1 and 2;

[0088] R 10 Selected from C3-C 10 Cycloalkyl, C6-C 10 Aryl, 5-10 membered heteroaryl and 3-10 membered heterocyclic group, the C3-C 10 Cycloalkyl, C6-C 10 Aryl, 5-10 membered heteroaryl or 3-10 membered heterocyclyl are each optionally substituted by one or more R 14 replace;

[0089] R 11Each is independently selected from C1-C6 alkyl, oxo, 5-10 membered heteroaryl, 3-10 membered heterocyclyl, R6C(O)-, R6C(O)N(R 12 )- and -NR7R8, wherein the C1-C6 alkyl, 5-10 membered heteroaryl or 3-10 membered heterocyclyl are each optionally substituted by one or more R 15 replace;

[0090] R 12 、R 13 Each independently selected from H, C1-C6 alkyl and C3-C 10 Cycloalkyl;

[0091] R 14 are each independently selected from H, halogen, cyano, oxo, C1-C6 alkyl, C1-C6 alkoxy, -CONR7R8 and R6C(O)-; and

[0092] R 15 Each independently selected from H, halogen, cyano, oxo, C1-C6 alkyl, C3-C 10 Cycloalkyl and 3-10 membered heterocyclic groups, wherein the C1-C6 alkyl, C3-C 10 The cycloalkyl or 3-10 membered heterocyclyl are each optionally substituted with one or more substituents selected from C1-C6 alkyl and oxo; and

[0093] x, y, z are each independently selected from 1, 2 and 3.

[0094] According to some embodiments of the present invention, ring A is selected from 4-10 membered heterocyclyl, C4-C 10 Cycloalkyl, C6-C 10 aryl and 5-10 membered heteroaryl.

[0095] According to some embodiments of the present invention, at most one of X1, X2, and X3 is N.

[0096] According to some embodiments of the present invention, ring A is a 5-8 membered heterocyclic group, preferably a 6-7 membered heterocyclic group, and more preferably a 6-7 membered nitrogen-containing heterocyclic group.

[0097] In some embodiments of the invention, Ring A is in,

[0098] X4 is selected from O and NH; d is 0 or 1; e is 1 or 2; wavy line It indicates the point of attachment of the group to the rest of the molecule;

[0099] Symbol 1 represents ring A and and symbol 2 represents the connection point between ring A and ring B.

[0100] In some embodiments of the present invention, Ring A is selected from Among them, the wavy line It indicates the point of attachment of the group to the rest of the molecule;

[0101] Symbol 1 represents ring A and and symbol 2 represents the connection point between ring A and ring B.

[0102] According to some embodiments of the present invention, Ring B is a 5-6 membered heterocycle, more preferably a 5-6 membered nitrogen-containing heterocycle.

[0103] In some embodiments of the invention, Ring B is The wavy line Indicates the connection point between ring B and ring A.

[0104] According to some embodiments of the invention, R1 is H.

[0105] According to some embodiments of the present invention, R2 is selected from H and C1-C6 haloalkyl, preferably C1-C6 haloalkyl, further preferably C1-C3 haloalkyl, and more preferably trifluoromethyl.

[0106] According to some embodiments of the present invention, R3 and R4 are each independently selected from H, oxo, C1-C6 alkyl, C3-C 10 Cycloalkyl, 3-10 membered heterocyclic group and R6C(O)-, wherein said C1-C6 alkyl, C3-C 10 cycloalkyl or 3-10 membered heterocyclyl are each optionally substituted with one or more R9;

[0107] R6 is selected from C1-C6 alkyl and C3-C 10 cycloalkyl; and R9 are each independently selected from halogen and C1-C6 alkyl.

[0108] In some embodiments of the present invention, R3 and R4 are each independently selected from H, oxo, C1-C6 alkyl, C3-C6 cycloalkyl, 4-6 membered heterocyclyl and R6C(O)-, wherein the C1-C6 alkyl is optionally substituted by 1, 2, 3, 4 or 5 halogens, and the 4-6 membered heterocyclyl is optionally substituted by 1, 2 or 3 C1-C6 alkyl; and R6 is C1-C6 alkyl.

[0109] In some embodiments of the present invention, R3, R4 are each independently selected from H, oxo, C1-C6 alkyl, C3-C6 cycloalkyl, oxetanyl, piperidinyl, tetrahydropyranyl and R6C(O)-, wherein the C1-C6 alkyl is optionally substituted by 1, 2, 3, 4 or 5 halogens, and the piperidinyl or tetrahydropyranyl are each optionally substituted by 1 or 2 C1-C6 alkyls; and R6 is C1-C6 alkyl.

[0110] In some embodiments of the present invention, R3 and R4 are each independently selected from H, oxo, methyl, isopropyl, cyclopropyl, cyclobutyl, CF3CH2-, CH3CO-, The wavy line Indicates the point of attachment of a group to the rest of the molecule.

[0111] In some embodiments of the present invention, R3 is selected from H, oxo and C1-C6 alkyl, preferably selected from H, oxo and C1-C3 alkyl, more preferably selected from H, oxo and methyl.

[0112] In some embodiments of the present invention, R4 is selected from C1-C6 alkyl, C3-C6 cycloalkyl, 4-6 membered heterocyclyl and R6C(O)-, wherein the C1-C6 alkyl is optionally substituted with 1, 2, 3, 4 or 5 halogens, and the 4-6 membered heterocyclyl is optionally substituted with 1, 2 or 3 C1-C6 alkyl; and R6 is C1-C6 alkyl.

[0113] In some embodiments of the present invention, R4 is selected from H, C1-C6 alkyl, C3-C6 cycloalkyl, oxetanyl, piperidinyl, tetrahydropyranyl and R6C(O)-, wherein the C1-C6 alkyl is optionally substituted with 1, 2, 3, 4 or 5 halogens, and the piperidinyl or tetrahydropyranyl are each optionally substituted with 1 or 2 C1-C6 alkyl groups; and R6 is C1-C6 alkyl.

[0114] In some embodiments of the present invention, R4 is selected from C1-C6 alkyl, C3-C6 cycloalkyl, oxetanyl, piperidinyl, tetrahydropyranyl and R6C(O)-, wherein the C1-C6 alkyl is optionally substituted with 1, 2, 3, 4 or 5 halogens, and each of the piperidinyl or tetrahydropyranyl is optionally substituted with 1 or 2 C1-C6 alkyl groups; and

[0115] R6 is a C1-C6 alkyl group.

[0116] In some embodiments of the present invention, R4 is selected from methyl, isopropyl, cyclopropyl, cyclobutyl, CF3CH2-, CH3CO-, The wavy line Indicates the point of attachment of a group to the rest of the molecule.

[0117] According to some embodiments of the present invention, x, y, z are each independently selected from 1 and 2.

[0118] According to some embodiments of the present invention,

[0119] R is selected from 3-12 membered heterocyclyl, 5-10 membered heteroaryl and -LR 10 wherein the 3-12 membered heterocyclyl or 5-10 membered heteroaryl are each optionally substituted by one or more R 11 replace;

[0120] L is -(L1) m -(L2) n -(L3) p -;

[0121] L1 is selected from -O-, -NH- and -CH2-O-;

[0122] L2 is selected from C1-C6 alkylene, C3-C 10 Cycloalkylene and 3-10 membered heterocyclylene;

[0123] L3 is selected from -NH-, -NHC(O)- and -C(O)-;

[0124] m, n and p are each independently selected from 0 and 1, and m, n and p are not 0 at the same time;

[0125] R 10 Selected from C3-C 10 Cycloalkyl, C6-C 10 Aryl, 5-10 membered heteroaryl and 3-10 membered heterocyclic group, the C3-C 10 Cycloalkyl, C6-C 10 Aryl, 5-10 membered heteroaryl or 3-10 membered heterocyclyl are each optionally substituted by one or more R 14 replace;

[0126] R 11 Each is independently selected from C1-C6 alkyl, oxo, 5-10 membered heteroaryl, 3-10 membered heterocyclyl and R6C(O)-, wherein the 3-10 membered heterocyclyl is optionally replaced by one or more R 15 replace;

[0127] R 14 are each independently selected from H, halogen, cyano, oxo and C1-C6 alkyl; and

[0128] R 15 Each is independently selected from oxo and C1-C6 alkyl.

[0129] According to some embodiments of the present invention,

[0130] L is -(L1) m -(L2) n -(L3) p -*, where the bond marked with "*" is connected to R 10 ;

[0131] L1 is selected from -O-, -N(R 12 )-、-C(R 12 )(R 13 )-O-@、-N(R 12 )CO-@ and -S-, wherein the bond marked with "@" is connected to L2;

[0132] L2 is selected from C1-C6 alkylene, C1-C6 heteroalkylene, C3-C 10 Cycloalkylene and 3-10 membered heterocyclylene;

[0133] L3 is selected from -N(R 12 )-、-N(R 12 )C(O)-$ and -C(O)-, where the bond marked with "$" is connected to R 10 ;

[0134] m, n and p are each independently selected from 0 and 1, and m, n and p are not 0 at the same time;

[0135] Preferably, L1 is selected from -O-, -N(R 12 )-;

[0136] L2 is selected from C1-C6 alkylene, C3-C6 cycloalkylene and 3-6 membered heterocyclylene;

[0137] L3 is selected from -N(R 12 )-、-N(R 12 )C(O)-$ and -C(O)-, more preferably -C(O)-, wherein the bond identified by "$" is connected to R 10 ;

[0138] Preferably, L1 is selected from -O-, -NH-;

[0139] L2 is selected from ethylene, propylene, cyclohexylene, and piperidinylene;

[0140] L3 is selected from -NH-, -NH-C(O)-$, -C(O)-, wherein the bond marked with "$" is connected to R 10 ;

[0141] Preferably, L is -O-, -O-cyclohexylene-*, -NH-cyclohexylene-*, -NH-piperidinylene-C(O)-*, -O-propylene-*, -NH-propylene-*, -O-ethylene-NH-*, -ethylene-NHC(O)-*, -NH-propylene-C(O)-* and -O-propylene-C(O)-*, wherein the bond marked with "*" is connected to R 10 ;

[0142] More preferably, L is selected from -O-propylene-*, -O-cyclohexylene-*, -NH-propylene-*, -NH-cyclohexylene-*, -NH-piperidinylene-C(O)-*, wherein the bond marked with "*" is connected to R 10 .

[0143] According to some embodiments of the present invention, the compound of the present invention is a compound of formula 1':

[0144] wherein R, R2, R3, R4, Ring A and x are as defined above for the compound of formula (1).

[0145] According to some embodiments of the present invention, the compound of the present invention is a compound of Formula 1A, 1B, IC or ID:

[0146] wherein R and R4 are as defined above for the compound of formula (1).

[0147] According to some embodiments of the present invention, the compound of the present invention is a compound of Formula 1A-1:

[0148] Among them, R 11 , R4 is as defined above for the compound of formula (1); Ring C is selected from 3-12 membered heterocyclyl and 5-10 membered heteroaryl; and r is selected from 0, 1 and 2.

[0149] In some embodiments of the present invention, in the compound of Formula 1A-1, Ring C is selected from a 3-12 membered heterocyclyl.

[0150] In some embodiments of the present invention, in the compound of Formula 1A-1:

[0151] Ring C is a 5-12 membered spiro heterocyclic group or a 5-12 membered bridged heterocyclic group, preferably a 7-12 membered spiro heterocyclic group or a 7-12 membered bridged heterocyclic group, more preferably More preferably and

[0152] R 11Each independently selected from C1-C6 alkyl and oxo, preferably, R 11 each independently selected from methyl and oxo,

[0153] The wavy line Indicates the point of attachment of a group to the rest of the molecule.

[0154] In some embodiments of the present invention, Ring C is More preferably and

[0155] R 11 Each independently selected from C1-C6 alkyl and oxo, preferably, R 11 each independently selected from methyl and oxo,

[0156] The wavy line Indicates the point of attachment of a group to the rest of the molecule.

[0157] In some embodiments of the present invention, in the compound of Formula 1A-1:

[0158] Ring C is a 3-10 membered monocyclic heterocyclic group or a 5-10 membered heteroaryl group, preferably a 6-7 membered monocyclic heterocyclic group or a 5-6 membered heteroaryl group, more preferably a piperidinyl group, a piperazinyl group, an azepanyl group, a 1,4-diazepanyl group or a pyrazolyl group;

[0159] R 11 Each is independently selected from C1-C6 alkyl, oxo, 5-6 membered heteroaryl, 5-7 membered heterocyclyl and R6C(O)-, wherein the C1-C6 alkyl, 5-6 membered heteroaryl or 5-7 membered heterocyclyl are each optionally substituted by 1, 2, 3 or 4 R 15 replace;

[0160] R6 is C4-C6 cycloalkyl; and R 15 are each independently selected from oxo and C1-C6 alkyl,

[0161] Preferably, ring C is a 3-10 membered monocyclic heterocyclic group, preferably a 6-7 membered monocyclic heterocyclic group, more preferably a piperidinyl group, a piperazinyl group, an azepanyl group, or a 1,4-diazepanyl group;

[0162] Preferably, R 11 Each is independently selected from C1-C6 alkyl, oxo, pyrazolyl, piperidinyl and C4-C6 cycloalkyl-C(O)-, wherein the piperidinyl is optionally substituted with 1 or 2 substituents selected from C1-C6 alkyl and oxo;

[0163] More preferably, R 11 Each is independently selected from methyl, oxo, pyrazolyl, N-methylpiperidinyl, oxopiperidinyl and cyclobutylcarbonyl.

[0164] According to some embodiments of the present invention, the compound of the present invention is a compound of Formula 1A-1a, 1A-1b, or 1A-1c:

[0165] wherein R4 is as defined above for the compound of formula (1); t is selected from 0, 1, 2 and 3; u, v, w, a, b and c are each independently selected from 1, 2 and 3; Y is selected from -CH- and -N-; and r is selected from 1 and 2.

[0166] According to some embodiments of the present invention, the compound of the present invention is a compound of Formula 1A-2:

[0167] wherein R4 is as defined above for the compound of formula (1);

[0168] L is -(L1) m -(L2) n -(L3) p -, preferably, L is -(L1) m -(L2) n -(L3) p -*, wherein the bond identified by "*" is connected to ring D;

[0169] L1 is selected from -O-, -NH- and -CH2-O-. Preferably, when L1 is -CH2-O-, the oxygen of -CH2-O- is connected to L2. Preferably, L1 is selected from -O-, -NH-;

[0170] L2 is selected from C1-C6 alkylene, C3-C 10 Cycloalkylene and 3-10 membered heterocyclylene, preferably selected from C1-C6 alkylene, C3-C6 cycloalkylene and 3-6 membered heterocyclylene, preferably selected from C1-C6 alkylene and C3-C6 cycloalkylene, preferably selected from ethylene, propylene, cyclohexylene and piperidinylene, more preferably selected from ethylene, propylene and cyclohexylene;

[0171] L3 is selected from -NH-, -NHC(O)- and -C(O)-. Preferably, when L3 is -NHC(O)-, the amino portion of the -NHC(O)- is connected to L2, and the carbonyl portion is connected to ring D;

[0172] m, n and p are each independently selected from 0 and 1, and m, n and p are not 0 at the same time;

[0173] Preferably, L is selected from -O-, -O-cyclohexylene-, -NH-cyclohexylene-, -NH-piperidinylene-, -O-propylene-, -NH-propylene-, -ethylene-, -ethylene-NHC(O)-, and -O-propylene-C(O)-;

[0174] Preferably, L is selected from -O-, -O-cyclohexylene-, -NH-cyclohexylene-, -NH-piperidinylene-C(O)-, -O-propylene-, -NH-propylene-, -O-ethylene-NH-, -ethylene-NHC(O)-, -NH-propylene-C(O)-, and -O-propylene-C(O)-;

[0175] Preferably, L is selected from -O-, -O-cyclohexylene-*, -NH-cyclohexylene-*, -NH-piperidinylene-C(O)-*, -O-propylene-*, -NH-propylene-*, -O-ethylene-NH-*, -ethylene-NHC(O)-*, -NH-propylene-C(O)-*, and -O-propylene-C(O)-*, wherein the bond marked with "*" is connected to ring D;

[0176] More preferably, L is selected from -O-propylene-*, -O-cyclohexylene-*, -NH-propylene-*, -NH-cyclohexylene-*, -NH-piperidinylene-C(O)-*, wherein the bond marked with "*" is connected to ring D;

[0177] Ring D is selected from C3-C 10 Cycloalkyl, C6-C 10 Aryl, 5-10 membered heteroaryl and 3-10 membered heterocyclic group, preferably selected from C3-C6 cycloalkyl, C6-C 10 Aryl, 5-6 membered heteroaryl and 5-7 membered heterocyclyl, preferably selected from cyclobutyl, phenyl, pyrazolyl, piperidinyl, piperazinyl, pyrrolidinyl, imidazolidinyl, morpholinyl, 1,3-oxazinyl, azepanyl and 1,4-oxazepanyl;

[0178] R 14 are each independently selected from H, halogen, cyano, oxo and C1-C6 alkyl; and

[0179] s is selected from 0, 1 and 2.

[0180] According to some embodiments of the present invention, the compound of the present invention is a compound of Formula 1A-2a, 1A-2b, 1A-2c, 1A-2d, 1A-2e, or 1A-2f:

[0181] Among them, R4, R 14As defined above for compounds of formula (1A-2); X is selected from -O-, -NH- and -CH2-O-, preferably, X is selected from -O- and -NH-; V is selected from -O-, -S-, -NR 12 -and-CHR 12 -, preferably, V is selected from -O- and -CHR 12 -;R 12 is selected from H and C1-C6 alkyl; h and l are each independently selected from 0, 1, 2 and 3; i is selected from 0, 1 and 2; and j and k are each independently selected from 1, 2 and 3.

[0182] According to some embodiments of the present invention, the compound of the present invention is a compound of Formula 1B-2a, 1C-2a or 1D-2a:

[0183] wherein R4 is as defined above for the compound of formula (1); X is selected from -O-, -NH- and -CH2-O-, preferably, X is selected from -O- and -NH-, more preferably, X is selected from -O-; V is selected from -O-, -S-, -NR 12 -and-CHR 12 -, preferably, V is selected from -CHR 12 -;R 12 is selected from H and C1-C6 alkyl; h and l are each independently selected from 0, 1, 2 and 3; and i is selected from 0, 1 and 2.

[0184] According to some embodiments of the invention, the compound of the invention is selected from the group consisting of:

[0185] Preparation method

[0186] Another object of the present invention is to provide a method for preparing the compound of the present invention.

[0187] It includes Route A or Route B as follows:

[0188] wherein R1, R2, R3, R4, x, y, z, X1, X2 and X3 are as defined above.

[0189] Pharmaceutical compositions and kits

[0190] Another object of the present invention is to provide a pharmaceutical composition comprising an effective amount of a compound of the present invention or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, hydrate, N-oxide, isotope-labeled compound, metabolite, ester, or prodrug thereof, and one or more pharmaceutically acceptable carriers.

[0191] Another object of the present invention is to provide a kit comprising an effective amount of a compound of the present invention or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, hydrate, N-oxide, isotopically labeled compound, metabolite, ester, or prodrug thereof, or a pharmaceutical composition of the present invention, and optionally a package insert.

[0192] In the present invention, "pharmaceutically acceptable carrier" refers to a diluent, adjuvant, excipient or vehicle that is administered together with the therapeutic agent and is suitable for contact with the tissues of humans and / or other animals without excessive toxicity, irritation, allergic response or other problems or complications corresponding to a reasonable benefit / risk ratio within the scope of reasonable medical judgment.

[0193] Pharmaceutically acceptable carriers that can be used in the pharmaceutical compositions or pharmaceutical formulations of this invention include, but are not limited to, sterile liquids, such as water and oils, including those of petroleum, animal, vegetable or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil and the like.

[0194] The pharmaceutical composition can be in the form of a solid preparation, a semisolid preparation, a liquid preparation, or a gaseous preparation. Examples of solid preparations include tablets, capsules, powders, granules, or suppositories, and examples of liquid preparations include solutions, suspensions, or injections. The composition can also be in the form of liposomes, microspheres, or other dosage forms. In particular, the pharmaceutical composition is in the form of a preparation suitable for oral administration.

[0195] When the pharmaceutical composition is administered intravenously, water is an exemplary carrier. Physiological saline and glucose and glycerol aqueous solutions can also be used as liquid carriers, particularly for injections. Suitable pharmaceutical excipients include starch, glucose, lactose, sucrose, gelatin, maltose, chalk, silica gel, sodium stearate, glyceryl monostearate, talc, sodium chloride, skim milk powder, glycerol, propylene glycol, water, ethanol, etc. The composition can also optionally include a small amount of wetting agent, emulsifier or pH buffer. Oral formulations can include standard carriers, such as pharmaceutical grade mannitol, lactose, starch, magnesium stearate, saccharin sodium, cellulose, magnesium carbonate, etc. Examples of suitable pharmaceutically acceptable carriers are described in Remington's Pharmaceutical Sciences (1990).

[0196] The pharmaceutical compositions of the present invention can act systemically and / or locally. For this purpose, they can be administered by suitable routes, for example, by injection (such as intravenous, intraarterial, subcutaneous, intraperitoneal, intramuscular injection, including instillation) or transdermal administration; or by oral, buccal, nasal, transmucosal, topical, in the form of ophthalmic preparations or by inhalation.

[0197] For these routes of administration, the pharmaceutical compositions of the present invention can be administered in suitable dosage forms, including but not limited to tablets, capsules, lozenges, hard candies, powders, sprays, creams, ointments, suppositories, gels, pastes, lotions, ointments, aqueous suspensions, injectable solutions, elixirs, syrups, and the like.

[0198] The content or dosage of the compound of the present invention in the pharmaceutical composition may be about 0.001 mg to about 1000 mg, suitably 0.001-100 mg, preferably 0.001-10 mg, more preferably 0.05-5 mg, particularly preferably 0.01-1 mg.

[0199] In some embodiments, the present invention provides a method for preparing a pharmaceutical composition of the present invention, comprising combining a compound of the present invention or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, hydrate, N-oxide, isotopically labeled compound, metabolite, ester, or prodrug thereof with one or more pharmaceutically acceptable carriers.

[0200] Treatment methods and uses

[0201] Another object of the present invention is to provide a method for treating a ULK1-mediated disease in a subject in need thereof, comprising administering to the subject an effective amount of a compound of the present invention or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, hydrate, N-oxide, isotope-labeled compound, metabolite, ester, or prodrug thereof, or a pharmaceutical composition of the present invention.

[0202] Another object of the present invention is to provide a compound of the present invention or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, hydrate, N-oxide, isotope-labeled compound, metabolite, ester, or prodrug thereof, or a pharmaceutical composition of the present invention for preparing a medicament for treating a ULK1-mediated disease in a subject in need thereof.

[0203] Another object of the present invention is to provide a compound of the present invention or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, hydrate, N-oxide, isotope-labeled compound, metabolite, ester, or prodrug thereof, or a pharmaceutical composition of the present invention, for use in treating a ULK1-mediated disease in a subject in need thereof.

[0204] Another object of the present invention is to provide a method for preventing or treating ULK1-mediated related diseases, which comprises administering to an individual in need thereof a preventively or therapeutically effective amount of a compound of the present invention or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotope-labeled compound, metabolite or prodrug thereof, or a pharmaceutical composition of the present invention.

[0205] According to some embodiments of the invention, the ULK1-mediated disease is characterized by aberrant autophagy.

[0206] According to some embodiments of the invention, the abnormal autophagy is induced by a treatment.

[0207] According to some embodiments of the invention, the ULK1-mediated disease is cancer, such as lung cancer, breast cancer or pancreatic cancer, in particular, the lung cancer is non-small cell lung cancer, the breast cancer is triple-negative breast cancer, or the pancreatic cancer is pancreatic ductal adenocarcinoma.

[0208] According to some embodiments of the invention, the ULK1 -mediated disease is tuberous sclerosis complex (TSC) or lymphangioleiomyomatosis (LAM).

[0209] According to some embodiments of the invention, the medicament is co-administered with an additional therapeutic agent.

[0210] According to some embodiments of the invention, the additional therapeutic agent is a standard of care therapy.

[0211] According to some embodiments of the invention, the standard of care therapy is an mTOR inhibitor, carboplatin, a MEK inhibitor, or a PARP inhibitor.

[0212] According to some embodiments of the invention, wherein the medicament is for degrading ATG13 in the subject.

[0213] As used herein, the term "effective amount" refers to an amount sufficient to achieve the desired prophylactic or therapeutic effect, for example, to achieve relief of one or more symptoms associated with the disease being treated.

[0214] The dosage regimen can be adjusted to provide the optimal desired response. For example, a single bolus can be administered, several divided doses can be administered over time, or the dose can be proportionally reduced or increased as indicated by the urgency of the therapeutic situation. It is to be noted that dosage values ​​can vary with the type and severity of the condition to be alleviated and can include single or multiple doses. It is to be further understood that for any particular individual, the specific dosage regimen should be adjusted over time according to the individual's needs and the professional judgment of the person administering or supervising the administration of the composition.

[0215] The amount of the compound of the present invention administered will depend on the severity of the individual, disease or the patient's condition, the speed of administration, the disposal of the compound and the judgment of the prescribing physician for treatment. Generally speaking, effective dose is about 0.0001 to about 50mg per kg body weight per day, for example, about 0.01 to about 10mg / kg / day (single or divided administration). For 70kg people, this will add up to about 0.007mg / day to about 3500mg / day, for example, about 0.7mg / day to about 700mg / day. In some cases, it can be enough to be not higher than the dosage level of the lower limit of the aforementioned range, and in other cases, it is still possible to adopt a larger dose in the case of not causing any harmful side effects, provided that the larger dose is first divided into several smaller doses to be administered throughout the day.

[0216] As used herein, unless otherwise indicated, the terms "treat," ...

[0217] As used herein, "subject" includes humans and non-human animals. Exemplary human subjects include human subjects suffering from diseases (e.g., the diseases described herein) (referred to as patients) or normal individuals. "Non-human animals" herein include all vertebrates, such as non-mammals (e.g., birds, amphibians, reptiles) and mammals, such as non-human primates, livestock and / or domesticated animals (e.g., sheep, dogs, cats, cows, pigs, etc.). Example

[0218] In order to make the purpose and technical scheme of the present invention clearer, the embodiments of the present invention are described in detail below in conjunction with embodiment.But those skilled in the art will understand that the following examples are only used to illustrate the present invention and should not be regarded as limiting the scope of the present invention.Unindicated specific conditions in the examples are all carried out according to the conditions of normal conditions or manufacturer's advice.Reagents used or instruments not indicated by manufacturer are all conventional products that can be obtained commercially.

[0219] The structures of the compounds were determined by nuclear magnetic resonance (NMR) and / or mass spectrometry (MS). -6 The unit of ppm is given. NMR measurements were performed using a Bruker AVANCE NEO 500M NMR spectrometer. The solvents used were deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), and deuterated methanol (CD3OD), and tetramethylsilane (TMS) was used as the internal standard.

[0220] MS was measured using an Agilent 1200 / 1290 DAD-6110 / 6120 Quadrupole MS liquid spectrometer / mass spectrometer (manufacturer: Agilent, MS model: 6110 / 6120 Quadrupole MS), a Waters ACQuity UPLC-QD / SQD (manufacturer: Waters, MS model: Waters ACQuity Qda Detector / Waters SQ Detector), and a THERMO Ultimate 3000-Q Exactive (manufacturer: THERMO, MS model: THERMO Q Exactive).

[0221] High performance liquid chromatography (HPLC) analysis was performed using Agilent HPLC 1200DAD, Agilent HPLC 1200VWD and Waters HPLC e2695-2489 liquid chromatographs.

[0222] Chiral HPLC analysis was performed using an Agilent 1260DAD high performance liquid chromatograph.

[0223] High performance liquid chromatography (HPLC) was performed using Waters 2767, Waters 2767-SQ Detecor2, Shimadzu LC-20AP, and Gilson-281 preparative chromatographs.

[0224] Chiral preparations were performed using a Shimadzu LC-20AP preparative chromatograph.

[0225] The CombiFlash rapid preparation instrument used was Combiflash Rf200 (TELEDYNE ISCO).

[0226] The thin layer chromatography silica gel plate used was Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plate, and the silica gel plate used in thin layer chromatography (TLC) had a specification of 0.15 mm to 0.2 mm.

[0227] Silica gel column chromatography generally uses Yantai Huanghai 200-300 mesh silica gel as the carrier.

[0228] The average kinase inhibition rate and IC50 value were determined using a NovoStar microplate reader (BMG, Germany).

[0229] The known starting materials disclosed herein can be synthesized by methods known in the art, or can be purchased from ABCR GmbH & Co. KG, Acros Organics, Aldrich Chemical Company, Accela ChemBio Inc, Darui Chemicals, and other companies.

[0230] A CEM Discover-S 908860 microwave reactor was used for the microwave reaction.

[0231] Unless otherwise specified in the examples, the solution refers to an aqueous solution.

[0232] Unless otherwise specified in the examples, the reaction temperature is room temperature, 20°C to 30°C.

[0233] The reaction progress in the examples was monitored by thin layer chromatography (TLC). The developing solvent used in the reaction, the eluent system for column chromatography used to purify the compound, and the developing solvent system for thin layer chromatography included: A: dichloromethane / methanol system, B: n-hexane / ethyl acetate. The volume ratio of the solvents was adjusted according to the polarity of the compound, and a small amount of alkaline or acidic reagents such as triethylamine and acetic acid could also be added for adjustment.

[0234] Preparation of intermediate Int-1

[0235] Step 1: Compound Int-1a (8.00 g, 36.99 mmol, Bidex) was dissolved in tetrahydrofuran (100 mL) at room temperature. Lithium aluminum tetrahydride (48.09 mL, 48.09 mmol, Anergy) was added dropwise under ice-cooling. The mixture was reacted at 65°C for 18 hours. The reaction solution was quenched with sodium sulfate decahydrate under ice-cooling. The mixture was filtered, and the filtrate was concentrated under reduced pressure to obtain compound Int-1b. MS m / z (ESI): 131.0 [M+1].

[0236] Step 2: To a solution of compound Int-1b (3.00 g, 23.04 mmol) in dioxane (50 mL) at room temperature, triethylamine (7.00 g, 69.13 mmol) and compound Int-1c (4.59 g, 23.04 mmol, Bid) were added, respectively. The atmosphere was purged with nitrogen three times, and the mixture was stirred at 110°C for 24 hours. The reaction mixture was concentrated under reduced pressure to obtain the crude product, which was then slurried with petroleum ether:ethyl acetate (2:1, 90 mL). The solid was washed with petroleum ether:ethyl acetate (2:1, 20 mL) and dried to obtain compound Int-1d. MS m / z (ESI): 278.0 [M+1].

[0237] Step 3: To a solution of compound Int-1d (3.40 g, 12.26 mmol) in tetrahydrofuran (30 mL) was added sodium borohydride (1.39 g, 36.79 mmol) and lithium borohydride (0.03 g, 1.23 mmol) under ice-cooling. The mixture was stirred at room temperature for 18 hours. The reaction mixture was poured into ice water (30 mL) and extracted with ethyl acetate (50 mL x 2). The mixture was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to afford the crude product. The crude product was purified by silica gel chromatography (eluent: MeOH:DCM = 0% to 8%) to afford compound Int-1e. MS m / z (ESI): 282.0 [M+1].

[0238] Step 4: Compound Int-1e (2.10 g, 7.47 mmol) was dissolved in 50% sulfuric acid (20 mL) at room temperature and stirred at 120°C for 1 hour to yield a black solution. The reaction mixture was poured into ice water (50 mL) and adjusted to pH 7-8 with saturated sodium bicarbonate solution. The mixture was extracted with dichloromethane (100 mL x 2), washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to yield the crude product. The crude product was purified by silica gel chromatography (eluent: DCM:MeOH = 0% to 8%) to yield compound Int-1f. MS m / z (ESI): 264.0 [M+1].

[0239] Step 5: To a solution of compound Int-1f (1.50 g, 5.70 mmol) in methanol (40 mL) was added palladium on carbon (60 mg, 0.57 mmol) at room temperature. The atmosphere was purged with hydrogen three times, and the reaction mixture was stirred under a hydrogen balloon at room temperature for 5 hours. The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure to yield compound Int-1. MS m / z (ESI): 234.1 [M+1].

[0240] Preparation of intermediate Int-2

[0241] Compound Int-1 (0.62 mL, 4.61 mmol, Bid) was added to a mixture of tert-butanol (5 mL) and 1,2-dichloroethane (40 mL). Zinc(II) chloride (10.14 mL, 10.14 mmol, 1 mol / L) was added at 0°C under nitrogen. The reaction was stirred at 0°C for 1 hour. Triethylamine (0.70 mL, 5.07 mmol) and compound Int-2a (1.08 g, 4.61 mmol) were then added, and the reaction was stirred at 0°C for 2 hours. The reaction mixture was concentrated, and the crude product was purified by thin-layer chromatography (developing solvent: DCM:MeOH = 100:1 to 10:1) to obtain compound Int-2. MS m / z (ESI): 414.9 [M+1].

[0242] Example 1. Preparation of Compound 1

[0243] At room temperature, compound Int-1 (120 mg, 0.51 mmol) was dissolved in dioxane (2 mL), and cesium carbonate (503 mg, 1.54 mmol), 43a (152 mg, 0.51 mmol), 1,1'-binaphthyl-2,2'-bis(diphenylphosphine) (64 mg, 0.10 mmol, Bidex) and palladium acetate (12 mg, 0.05 mol, Anaiji) were added to the reaction solution, and stirred at 100 ° C for 18 hours under nitrogen protection to obtain a suspension. The reaction solution was filtered and the filtrate was concentrated under reduced pressure to obtain a crude compound. The crude product was purified by preparative HPLC (chromatographic column: Waters-Xbridge-C18-10μm-19*250mm; mobile phase: A: 10mM ammonium bicarbonate / water; B: acetonitrile, gradient ratio: acetonitrile 95%, flow rate: 20 mL / min) to obtain compound 1. MS m / z(ESI):492.2[M+1]. 1 H NMR (400MHz, DMSO-d6): δ9.75 (s, 1H), 8.42 (s, 1H), 7.52 (d, J = 6.4Hz, 2H), 6.91 (d, J=9.0Hz,1H),4.78(d,J=11.8Hz,1H),4.56(d,J=11.8Hz,1H),4.11(s,2H),3.83(t ,J=5.2Hz,2H),3.70-3.55(m,2H),3.44(t,J=5.2Hz,2H),3.22-3.08(m,2H),2.89( d,J=8.8Hz,4H),2.74(d,J=11.0Hz,1H),2.64(d,J=9.4Hz,1H),2.36-2.19(m,5H).

[0244] Example 2. Preparation of Compound 2

[0245] To a solution of compound 44a (160 mg, 0.48 mmol) and compound Int-1 (123 mg, 0.53 mmol) in N,N-dimethylformamide (4 mL) was added hydrochloric acid (1.5 mL, 6.00 mmol, C = 4 mol / L) at 25°C. The mixture was heated to 100°C and stirred for 2 hours. The mixture was concentrated under reduced pressure to obtain a crude product. The crude product was purified by preparative HPLC (column: Waters-Xbridge-C18-10μm-19*250mm; mobile phase: A: 10 mM ammonium bicarbonate / water; B: acetonitrile, gradient: 40-60% acetonitrile, flow rate: 25 mL / min) to obtain compound 2. MS m / z (ESI): 532.2 [M+1]. 1 HNMR (400MHz, DMSO-d6): δ9.58(s,1H),8.34(s,1H),7.52(d,J=2.2Hz,2H),6.90(d ,J=9.0Hz,1H),4.78-4.72(m,1H),4.52(d,J=11.8Hz,1H),3.67-3.45(m,6H),3.20 -3.09(m,2H),2.88(dt,J=10.6,6.0Hz,1H),2.69(dd,J=10.2,3.0Hz,1H),2.60(s, 1H), 2.46 (d, J = 6.8Hz, 2H), 2.32 (s, 2H), 2.25 (d, J = 10.2Hz, 8H), 1.68-1.52 (m, 6H).

[0246] Example 3. Preparation of Compound 3

[0247] To a solution of compound 47a (135 mg, 0.40 mmol) and compound Int-1 (112 mg, 0.48 mmol) in isopropanol (5 mL) was added trifluoroacetic acid (300 μL, 4.00 mmol) at room temperature, and the mixture was heated to 40°C and stirred for 16 h. The mixture was concentrated under reduced pressure to yield a crude product. The crude product was purified by preparative HPLC (column: Waters-SunFire-C18-10 μm-19*250 mm; mobile phase: A: 0.1% formic acid / water; B: acetonitrile, gradient: acetonitrile 20-50%, flow rate: 25 mL / min) to yield compound 3. MS m / z (ESI): 535.3 [M+1]. 1H NMR (400MHz, DMSO-d6): δ10.00(s,1H),8.47(s,1H),7.75–7.59(m,1H),7.48(d,J=2.0Hz,1H),6 .95(d,J=8.6Hz,1H),4.77(d,J=11.8Hz,1H),4.56(d,J=11.8Hz,1H),4.42(t,J=6.4Hz,2H),3.6 8–3.55(m,2H),3.37(t,J=7.0Hz,2H),3.25–3.09(m,4H),2.94–2.85(m,1H),2.81–2.72(m,1H), 2.69–2.62(m,1H),2.36–2.24(m,5H),2.22–2.15(m,2H),2.00–1.88(m,2H),1.77–1.61(m,4H).

[0248] Example 4. Preparation of Compound 4

[0249] Step 1: To a solution of compound 65a (3.00 g, 14.91 mmol, Bid) and triethylamine (6.2 mL, 44.72 mmol) in dichloromethane (30 mL) was added methylsulfonyl chloride (1.73 mL, 22.36 mmol) at 0°C and stirred at 0°C for 3 hours. The mixture was poured into water (20 mL) and extracted with dichloromethane (30 mL x 3). The combined organic phases were washed with brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to yield compound 65b. MS m / z (ESI): 280.11 [M+1].

[0250] Step 2: To a solution of compound 65c (1.50 g, 22.03 mmol, Bid) in N,N-dimethylformamide (30 mL) at 0°C was added sodium hydride (1.32 g, 33.05 mmol, 60% purity). The mixture was stirred at 0°C for 20 minutes, followed by the addition of compound 65b (7.00 g, 13.78 mmol). The temperature was raised to 25°C and stirred for 18 hours. The mixture was poured into water (50 mL) for quenching and extracted with ethyl acetate (50 mL x 3). The organic phases were combined, washed with brine (30 mL x 4), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to yield the crude product. The crude product was purified by silica gel chromatography (eluent: petroleum ether-tetrahydrofuran = 5-45%) to yield compound 65d. MS m / z (ESI): 252.16 [M+1].

[0251] Step 3: To a solution of compound 65d (889 mg, 3.54 mmol) in 1,4-dioxane (15 mL) was added hydrochloric acid (10 mL, 40.00 mmol, 4 mol / L) at 25°C. The mixture was stirred at 25°C for 3 hours. The mixture was concentrated under reduced pressure to afford compound 65e. MS m / z (ESI): 152.16 [M+1].

[0252] Step 4: To a solution of compound Int-2 (110 mg, 0.12 mmol) in N,N-dimethylformamide (3 mL) at 25°C, compound 65e (27.40 mg, 0.18 mmol) and triethylamine (83.74 μL, 0.60 mmol) were added. The mixture was stirred at 100°C for 2 hours. The mixture was concentrated under reduced pressure to afford the crude product. The crude product was purified by preparative HPLC (column: Waters-Xbridge-C18-10 μm-19*250 mm; mobile phase: A: 10 mM ammonium bicarbonate / water; B: acetonitrile, gradient: 40-60% acetonitrile, flow rate: 25 mL / min) to afford compound 4. MS m / z (ESI): 529.26 [M+1]. 1 HNMR (400MHz, DMSO-d6): δ9.65(s,1H),8.39(s,1H),7.79(s,1H),7.55(s,2H),7.44(d,J=5.0Hz,1H),6. 90(d,J=9.0Hz,1H),6.23(d,J=1.4Hz,1H),4.74(d,J=11.8Hz,1H),4.56-4.46(m,2H),4.16(dd,J=13.2,0 .6Hz,2H),3.66-3.53(m,2H),3.26-3.06(m,4H),2.86(dd,J=9.2,6.6Hz,1H),2.69(dd,J=9.2,4.6Hz,1H ), 2.57 (dd, J = 10.0, 0.6Hz, 1H), 2.23 (d, J = 14.8Hz, 5H), 2.10 (dd, J = 11.8, 1.4Hz, 2H), 1.99-1.94 (m, 2H).

[0253] Example 5. Preparation of Compound 5

[0254] Compound Int-2 (50 mg, 0.30 mmol) and 79a (16.94 mg, 0.12 mmol, Bid) were dissolved in N,N-dimethylformamide (0.5 mL). N,N-Diisopropylethylamine (39.96 μL, 0.24 mmol) was added at room temperature and stirred for 1 hour. The reaction mixture was purified by HPLC using a Gilson 306-1741 column (Waters SunFire C18-10 μm-19*250 mm; mobile phase: water (containing 10 mM ammonium bicarbonate) and acetonitrile, gradient: acetonitrile 30% to 68%, flow rate: 20 mL / min) to afford compound 5. MS m / z (ESI): 518.4 [M+1]. 1 H NMR (400MHz, DMSO-d6): δ9.53(s,1H),8.29(s,1H),7.64–7.54(m,2H),6.90(d,J=8.6Hz,1H ),4.76(d,J=11.6Hz,1H),4.52(d,J=11.8Hz,1H),3.68–3.52(m,5H),3.47(s,1H),3.20–3.0 6(m,2H),2.86(s,1H),2.70–2.65(m,1H),2.58(d,J=10.6Hz,2H),2.45(d,J=8.6Hz,2H),2. 38–2.31(m,2H),2.22(d,J=4.6Hz,7H),1.93(dt,J=13.8,6.6Hz,2H),1.76(d,J=7.6Hz,2H).

[0255] Example 6. Preparation of Compound 6

[0256] Compounds Int-2 (50 mg, 0.12 mmol) and 83b (24.16 mg, 0.12 mmol) were dissolved in N,N-dimethylformamide (1 mL). N,N-Diisopropylethylamine (0.10 mL, 0.60 mmol) was added at room temperature, and the mixture was stirred at 100°C for 1 hour. The reaction mixture was purified by HPLC using a Gilson 306-1741 column (Waters SunFire C18-10 μm-19*250 mm; mobile phase: water (containing 0.1% formic acid) and acetonitrile, gradient: acetonitrile 14% to 95%, flow rate: 20 mL / min) to afford compound 6. MS m / z (ESI): 543.2 [M+1]. 1HNMR (400MHz, DMSO-d6): δ9.55(s,1H),8.17(s,1H),7.84(d,J=2.2Hz,1H),7.56(dd,J=6.4,2.8Hz ,2H),7.47(d,J=1.8Hz,1H),6.91(d,J=9.2Hz,1H),6.27(t,J=2.2Hz,2H),4.79(d,J=11.8Hz,1H),4 .55(d,J=11.8Hz,1H),4.29(d,J=19.2Hz,2H),3.71–3.55(m,2H),3.21–3.09(m,2H),2.87(d,J=8.4 Hz,1H),2.72(d,J=11.0Hz,1H),2.61(d,J=10.6Hz,1H),2.25(d,J=11.6Hz,7H),1.93–1.69(m,6H).

[0257] Example 7. Preparation of Compound 7

[0258] To a solution of compound 84a (100 mg, 0.16 mmol) and compound Int-1 (37.33 mg, 0.16 mmol) in isopropanol (3 mL) was added trifluoroacetic acid (0.12 mL, 1.60 mmol) at 25°C. The resulting mixture was heated to 40°C and stirred for 18 hours. After cooling to room temperature, the reaction solution was purified by HPLC using a Gilson 306-1741 column (Waters SunFire C18-10 μm-19*250 mm column); mobile phase: water (containing 0.1% formic acid) and acetonitrile, gradient: acetonitrile 0% to 95%, flow rate: 20 mL / min) to afford compound 7. MS m / z (ESI): 544.4 [M+1]. 1 H NMR (400MHz, DMSO-d6): δ10.01 (s, 1H), 8.53-8.44 (m, 1H), 7.67 (d, J = 2.2Hz, 1H), 7.59 -7.49(m,2H),7.45(d,J=1.8Hz,1H),6.94(d,J=8.6Hz,1H),6.24(t,J=2.0Hz,1H),5.43(s,1H),4.78(d,J=11.8Hz,1H),4.56(d,J=11.8Hz, 1H),4.34-4.28(m,1H),3.69-3.57(m,2H),3.23-3.11(m,3H),2.92-2 .90(m,1H),2.78-2.65(m,2H),2.32-2.28(m,4H),2.14-1.82(m,8H).

[0259] Example 8. Preparation of Compound 8

[0260] Compound Int-2 (40 mg, 0.10 mmol) and 85a (12 mg, 0.10 mmol) were dissolved in N,N-dimethylformamide (1.5 mL). N,N-Diisopropylethylamine (0.03 mL, 0.19 mmol) was added at room temperature and stirred for 1 hour. The reaction mixture was purified by HPLC using a Gilson 306-1741 column (Waters SunFire C18-10 μm-19 x 250 mm; mobile phase: water (containing 0.1% formic acid) and acetonitrile, gradient: acetonitrile 14% to 95%, flow rate: 20 mL / min) to afford compound 8. MS m / z (ESI): 503.4 [M+1]. 1 HNMR (400MHz, DMSO-d6): δ9.49(s,1H),8.15(s,1H),7.71(d,J=2.2Hz,1H),7.59(dd,J=8.6,2.4Hz,1H), 7.51(d,J=2.4Hz,1H),7.42(d,J=1.8Hz,1H),7.17(s,1H),6.90(d,J=8.6Hz,1H),6.21(t,J=2.0Hz,1H), 4.76(d,J=11.8Hz,1H),4.52(d,J=11.8Hz,1H),4.13(t,J=7.0Hz,2H),3.69–3.53(m,2H),3.43(q,J=6.4 Hz,2H),3.14(dd,J=26.0,10.6Hz,2H),2.85(s,1H),2.77–2.56(m,2H),2.23(s,5H),2.11-2.04(m,2H).

[0261] Example 9. Preparation of Compound 9

[0262] The first step: at 0 ° C, 2-chloro-2-oxoacetyl chloride (0.49 mL, 5.71 mmol) was added to a solution of 87a (400 mg, 2.28 mmol, Bid) in dichloromethane (3 mL) and stirred at 25 ° C for 1 hour under N2 atmosphere. The mixed solution was concentrated to dryness. The solid was then dissolved in tetrahydrofuran (3 mL) and sodium borohydride (0.84 mL, 22.80 mmol) was added. It was stirred at 25 ° C for 16 hours. The mixture was poured into water (10 mL) and quenched, and extracted with ethyl acetate (20 mL × 3). The organic phases were combined, washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated to dryness under reduced pressure to obtain a crude product. The crude product was purified by silica gel chromatography (eluent: petroleum ether: tetrahydrofuran = 1: 1) to give compound 87b. MS m / z (ESI): 162.2 [M + 1].

[0263] Step 2: To a solution of compound 87b (101 mg, 0.63 mmol) in tetrahydrofuran (5 mL) at 0°C was added sodium hydride (38.66 mg, 60% purity, 0.97 mmol) with stirring, followed by Int-2 (200 mg, 0.48 mmol). The mixture was stirred at 60°C under a nitrogen atmosphere for 18 hours. The mixture was quenched with water (1 mL) and concentrated under reduced pressure to afford the crude product. The crude product was purified by preparative HPLC (column: Waters-Xbridge-C18-10μm-19*250mm; mobile phase: A: 10 mM formic acid / water; B: acetonitrile, gradient: 40-60% acetonitrile, flow rate: 25 mL / min) to afford compound 9. MS m / z (ESI): 539.2 [M+1]. 1 H NMR (400MHz, DMSO-d6): δ10.01(s,1H),8.48(s,1H),7.78(dd,J=7.8,1.4Hz,1H) ,7.67–7.62(m,1H),7.57(dd,J=8.6,2.4Hz,1H),7.48–7.39(m,3H),6.91(d,J=8. 6Hz,1H),4.74(d,J=11.8Hz,1H),4.54–4.44(m,3H),3.66–3.56(m,2H),3.18–3. 09(m,2H),2.96–2.86(m,3H),2.72–2.57(m,2H),2.22(s,5H),2.15–2.08(m,2H).

[0264] Example 10. Preparation of Compound 10

[0265] Step 1: To a solution of compound 88a (4.30 g, 34.12 mmol, Bid) in dichloromethane (40 mL) at 0°C, tert-butyldimethylsilyl chloride (7.7 g, 51.19 mmol) and imidazole (4.60 g, 68.22 mmol) were added, respectively. The mixture was stirred at room temperature for 18 hours. The crude product was concentrated under reduced pressure and purified by silica gel chromatography (eluent: petroleum ether:ethyl acetate = 50:1-10:1) to afford compound 88b. MS m / z (ESI): 241.1 [M+1].

[0266] Step 2: To a solution of compound 88b (4.80 g, 20.0 mmol) in tetrahydrofuran (60 mL) at -78°C was added n-butyllithium (15.0 mL, 24.0 mmol). The mixture was stirred at -78°C for 1 hour. Hexachloroethane (7.10 g, 30.0 mmol) was then added, the temperature gradually returned to 25°C, and the mixture was stirred for 18 hours. The mixture was poured into water (100 mL) and extracted with ethyl acetate (50 mL x 3). The combined organic phases were washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to yield the crude product. The crude product was purified by silica gel chromatography (eluent: petroleum ether:ethyl acetate = 50:1-10:1) to yield compound 88c. MS m / z (ESI): 275.1 [M+1].

[0267] Step 3: To a solution of compound 88c (1.00 g, 3.63 mmol) in 1,4-dioxane (8 mL) was added hydrochloric acid / dioxane (2.00 mL, 8.00 mmol) at 0°C and stirred at room temperature for 2 hours. The mixture was adjusted to pH 7 with saturated sodium bicarbonate solution and extracted with dichloromethane (20 mL x 3). The combined organic phases were washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to yield compound 88d. MS m / z (ESI): 161.0 [M+1].

[0268] Step 4: To a solution of compound 88d (60 mg, 0.375 mmol) in tetrahydrofuran (5 mL) at 0°C was added sodium hydride (12 mg, 0.300 mmol, 60% purity). The mixture was stirred for 30 minutes, followed by the addition of compound Int-2 (100 mg, 0.250 mmol). The mixture was heated to 60°C and stirred for 18 hours. The mixture was concentrated under reduced pressure to afford the crude product. The crude product was purified by preparative HPLC (column: Waters-Xbridge-C18-10μm-19*250mm; mobile phase: A: 10 mM ammonium bicarbonate / water; B: acetonitrile, gradient: acetonitrile 35-95%, flow rate: 25 mL / min) to afford compound 10. MS m / z (ESI): 537.9 [M+1].1 H NMR (400MHz, DMSO-d6): δ10.02(s,1H),8.49(s,1H),7.61–7.52(m,2H),7.45(s,1H) ,6.92(d,J=8.6Hz,1H),6.39(d,J=1.8Hz,1H),4.75(d,J=11.8Hz,1H),4.54(d,J=11. 8Hz,1H),4.39(t,J=6.0Hz,2H),4.26(t,J=6.8Hz,2H),3.69–3.53(m,2H),3.22–3.07 (m,2H),2.93–2.85(m,1H),2.76–2.65(m,1H),2.63–2.56(m,1H),2.29–2.15(m,7H).

[0269] Example 11. Preparation of Compound 11

[0270] Step 1: To a solution of compound 89a (3.00 g, 14.91 mmol, Bid) in tetrahydrofuran (20 mL) at 25°C was added sodium hydride (0.61 g, 15.13 mmol, 60% purity). The reaction was stirred at 25°C for 20 minutes, followed by the addition of N-tert-butyloxycarbonyl-3-aminopropyl bromide (2643.00 mg, 11.10 mmol, Bid). The temperature was raised to 60°C and stirred for 18 hours. After cooling to room temperature, the mixture was quenched by pouring into water (30 mL) and extracted with ethyl acetate (50 mL x 3). The combined organic phases were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to afford the crude product. The crude product was purified by silica gel chromatography (eluent: petroleum ether-tetrahydrofuran = 50-60%) to afford compound 89b. MS m / z (ESI): 257.1 [M+1].

[0271] Step 2: To a solution of compound 89b (300 mg, 1.17 mmol) in dichloromethane (3 mL) was added hydrochloric acid (3 mL, 12.00 mmol, 4 mol / L) at 25°C. The mixture was stirred at 25°C for 3 hours. The mixture was concentrated under reduced pressure to afford compound 89c. MS m / z (ESI): 157.1 [M+1].

[0272] Step 3: To a solution of compound Int-2 (108 mg, 0.13 mmol) in acetonitrile (3 mL) at 25°C, compound 89c (24 mg, 0.16 mmol) and triethylamine (90.44 μL, 0.65 mmol) were added, respectively. The mixture was stirred at 90°C for 18 hours. The mixture was concentrated under reduced pressure to afford the crude product. The crude product was purified by preparative HPLC (column: Waters-Xbridge-C18-10 μm-19*250 mm; mobile phase: A: 10 mM ammonium bicarbonate / water; B: acetonitrile, gradient: 40-60% acetonitrile, flow rate: 25 mL / min) to afford compound 11. MS m / z (ESI): 534.4 [M+1]. 1 HNMR(400MHz,DMSO-d6): δ9.49(s,1H),8.18(s,1H),7.62-7.53(m,2H),7.19(t,J=5.8Hz,1H),6.91(d,J=8. 6Hz,1H),4.77(d,J=11.8Hz,1H),4.52(d,J=11.8Hz,1H),3.39(d,J=6.2Hz,2H),3. 31(t,J=6.6Hz,2H),3.27-3.01(m,6H),2.85(dt,J=7.2,3.6Hz,1H),2.72(dd,J=9. 0,5.6Hz,1H),2.62(dd,J=10.8,2.6Hz,1H),2.32-2.17(m,7H),1.79-1.63(m,6H).

[0273] Example 12. Preparation of Compound 12

[0274] Step 1: Under nitrogen, compound 92a (5.00 g, 23.12 mmol, Bidex) was dissolved in tetrahydrofuran (50 mL). Lithium aluminum hydride (36.99 mL, 92.47 mmol, Anergy) was then added in an ice bath. The mixture was slowly warmed to room temperature and stirred for 18 hours. After completion of the reaction, water (37 mL) and aqueous sodium hydroxide solution (37 mL, 15% wt) were added to the reaction solution. The mixture was stirred at room temperature for 15 minutes, followed by anhydrous sodium sulfate. The mixture was filtered, and the residue was washed with ethyl acetate (100 mL). The organic layers were separated and concentrated to dryness under reduced pressure to yield compound 92b. MS m / z (ESI): 131.1 [M+1].

[0275] Step 2: Compound 92b (2.00 g, 15.36 mmol) was dissolved in dimethyl sulfoxide (30 mL). Compound 92c (1.96 g, 12.29 mmol) and potassium hydroxide (2.15 g, 38.41 mmol, Anaiji) were then added, stirring at 60°C for 2 hours. After completion of the reaction, water (30 mL) was added to the reaction solution, followed by extraction with ethyl acetate (50 mL x 3). The combined organic phases were washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to dryness under reduced pressure. The residue was purified by silica gel column chromatography (methanol:dichloromethane = 2%-3%) to yield compound 92d. MS m / z (ESI): 250.1 [M+1].

[0276] Step 3: Compound 92d (300 mg, 1.2 mmol) was dissolved in methanol (5 mL), followed by the addition of wet palladium on carbon (128 mg, 0.12 mmol, 10%, Bidex). The mixture was stirred at room temperature for 2 hours under a hydrogen atmosphere. After completion of the reaction, the reaction mixture was filtered, washed with methanol, and the filtrate was concentrated to dryness under reduced pressure to obtain compound 92e. MS m / z (ESI): 220.0 [M+1].

[0277] Step 4: Compound 92e (150 mg, 0.68 mmol) was dissolved in isopropanol (1.5 mL), followed by the addition of compound 92f (184 mg, 0.54 mmol) and trifluoroacetic acid (254.88 μL, 3.42 mmol), and the mixture was stirred at 50°C for 18 hours. After completion of the reaction, the reaction solution was concentrated to dryness under reduced pressure, and the residue was purified by HPLC (Waters-Xbridge-C18-10 μm-19*250 mm column; mobile phase: water (containing 10 mmol NH4HCO3) and acetonitrile, gradient: acetonitrile 39% to 69%, flow rate: 25 mL / min) to afford compound 12. MS m / z (ESI): 521.0 [M+1]. 1H NMR (400MHz, DMSO-d6): δ9.90(s,1H),8.44(s,1H),7.13(d,J=7.2Hz,2H),6.82(d,J=8.8Hz,1H),4.41(t ,J=6.4Hz,2H),4.21(dd,J=10.4,2.8Hz,1H),3.94-3.81(m,1H),3.70-3.58(m,1H),3.39-3.34(m,2H),3. 24(t,J=5.6Hz,2H),3.02-2.94(m,1H),2.85(d,J=11.2Hz,1H),2.78(d,J=10.4Hz,1H),2.58(td,J=11.6, 3.2Hz, 1H), 2.21 (s, 3H), 2.18 (t, J = 6.4Hz, 2H), 2.11-2.03 (m, 1H), 1.97-1.92 (m, 2H), 1.73-1.63 (m, 5H).

[0278] Example 13. Preparation of Compound 13

[0279] To a solution of compound 95a (89 mg, 0.53 mmol, Bid) in acetonitrile (4 mL) were added triethylamine (0.20 mL, 1.45 mmol) and Int-2 (200 mg, 0.48 mmol), respectively, and the mixture was stirred at 25°C for 1 hour. The mixture was concentrated under reduced pressure to obtain a crude product. The crude product was purified by preparative HPLC (column: Waters-Xbridge-C18-10μm-19*250mm; mobile phase: A: 10 mM ammonium bicarbonate / water; B: acetonitrile, gradient: 40-60% acetonitrile, flow rate: 25 mL / min) to obtain compound 13. MS m / z (ESI): 546.4 [M+1]. 1 H NMR (400MHz, DMSO-d6): δ9.59(s,1H),8.33(s,1H),7.54(d,J=8.2Hz,2H),6.91(d,J=8.4Hz,1H),4.76(d,J=11.8Hz,1H),4.52(d,J =11.8Hz,1H),3.67–3.45(m,7H),3.19–3.09(m,2H),2.74–2.65(m,1H),2.62–2.56(m,1H),2.33–2.16(m,12H),1.51-1.47(m,8H).

[0280] Example 14. Preparation of Compound 14

[0281] Step 1: Compounds 99a (3.38 mL, 26.87 mmol, Bid) and 99b (7.81 g, 29.55 mmol, Bid) were dissolved in dimethyl sulfoxide (50 mL). Triethylamine (11.17 mL, 80.60 mmol) was added at room temperature and the mixture was heated to 60°C for 18 hours. The reaction mixture was diluted with water (50 mL) and adjusted to pH 5 with 1 M hydrochloric acid. The mixture was then extracted with ethyl acetate (30 mL x 3). The combined organic phases were washed with saturated brine (200 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to yield compound 99c. MS m / z (ESI): 431.3 [M+1].

[0282] Step 2: Compound 99c (1.20 g, 2.79 mmol) and iron powder (1.25 g, 22.31 mmol) were dissolved in acetic acid (30 mL) and reacted at 60°C for 3 hours. The reaction solution was diluted with water (50 mL) and adjusted to pH 7 with 2 M sodium hydroxide solution. The mixture was filtered, and the solid was washed with dichloromethane / methanol (5:1, 30 mL). The filtrate was concentrated to afford compound 99d. MS m / z (ESI): 353.4 [M+1].

[0283] Step 3: Dissolve compound 99d (666 mg, 1.89 mmol) and triethylamine (0.52 mL, 3.78 mmol) in tetrahydrofuran (5 mL). Add di-tert-butyl dicarbonate (0.49 mL, 2.27 mmol) and react at room temperature for 18 hours. The reaction solution was concentrated to dryness, and the crude product was purified by silica gel column chromatography (eluent: ethyl acetate:petroleum ether = 10:1 to 5:1) to obtain compound 99e. MS m / z (ESI): 453.6 [M+1].

[0284] Step 4: Compound 99e (230 mg, 0.51 mmol) was dissolved in tetrahydrofuran (5 mL). Sodium hydride (24 mg, 60% purity, 0.61 mmol) was added at 0°C and the mixture was allowed to react for 0.5 hours. Iodomethane (30.04 μL, 0.48 mmol) was added to the reaction mixture, and the mixture was allowed to warm to room temperature for 1 hour. The reaction mixture was diluted with water (20 mL) and extracted with ethyl acetate (30 mL x 3). The organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to dryness. The crude product was purified by silica gel column chromatography (eluent system: ethyl acetate:petroleum ether = 10:1 to 1:1) to obtain compound 99f. MS m / z (ESI): 467.5 [M+1].

[0285] Step 5: Dissolve compound 99f (220 mg, 0.33 mmol) in methanol (5 mL) and add 10% palladium on carbon (85 mg, 0.08 mmol). Incubate at room temperature under a hydrogen atmosphere for 18 hours. Filter the reaction mixture, and concentrate the filtrate to dryness to obtain compound 99g. MS m / z (ESI): 333.4 [M+1].

[0286] Step 6: Dissolve compound 99g (170 mg, 0.51 mmol) and formaldehyde (0.38 mL, 5.11 mmol) in methanol (5 mL). Add sodium triacetoxyborohydride (539 mg, 2.56 mmol) and react at room temperature for 4 hours. Quench with saturated sodium carbonate (20 mL) and extract with ethyl acetate (30 mL x 3). The combined organic phases are washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to yield compound 99h. MS m / z (ESI): 347.4 [M+1].

[0287] Step 7: Dissolve compound 99h (150 mg, 0.43 mmol) and trifluoroacetic acid (0.65 mL, 8.66 mmol) in dichloromethane (5 mL) and react at room temperature for 1 hour. Concentrate the reaction mixture to dryness to obtain compound 99i. MS m / z (ESI): 247.4 [M+1].

[0288] Step 8: Compounds 99i (50 mg, 0.20 mmol) and 78j (206 mg, 0.61 mmol) were dissolved in isopropanol (1 mL) and reacted at 40°C for 2 hours. The reaction solution was purified by HPLC (Gilson 306-1741, Shimadzu Shim-pack C18 5 μm-20 x 250 mm column; mobile phase: water (containing 0.1% formic acid) and acetonitrile, gradient: acetonitrile 14% to 95%, flow rate: 20 mL / min) to afford compound 14. MS m / z (ESI): 548.4 [M+1]. 1 H NMR (400MHz, DMSO-d6): δ10.03(s,1H),8.48(s,1H),7.51(d,J=2.4Hz,1H),7.34(s,1H),6.87(d,J=8.6Hz,1H),4.44(t,J=6.4Hz,2H),3.51(s,1 H), 3.25 (d, J = 10.4Hz, 8H), 2.89 (d, J = 11.2Hz, 1H), 2.68 (d, J = 11.2Hz, 1H), 2.28 (s, 3H), 2.17 (t, J = 6.0Hz, 2H), 2.09–1.92 (m, 5H), 1.68 (s, 4H).

[0289] Example 15. Preparation of Compound 15

[0290] Compound Int-1 (40 mg, 0.14 mmol) was dissolved in isopropanol (1 mL), followed by the addition of compound 100a (46 mg, 0.14 mmol) and trifluoroacetic acid (0.10 mL, 1.37 mmol), and the mixture was stirred at 40°C for 18 hours. After completion of the reaction, the reaction solution was purified by HPLC (Waters-SunFire-C18-10μm-19*250mm column; mobile phase: water (containing 0.1% formic acid) and acetonitrile, gradient: acetonitrile 12% to 52%, flow rate: 25 mL / min) to afford compound 15. MS m / z (ESI): 535.0 [M+1]. 1 H NMR (400MHz, DMSO-d6): δ9.96 (s, 1H), 8.47 (s, 1H), 7.30 (d, J = 8.8Hz, 1H), 7.20 -7.09(m,1H),6.91(d,J=8.8Hz,1H),4.46-4.37(m,3H),4.11-4.06(m,1H),3.4 0-3.36(s,2H),3.24(t,J=5.6Hz,3H),3.12-2.97(m,4H),2.65-2.60(m,1H),2. 26-2.20(m,4H),2.19-2.15(m,2H),2.13-2.07(m,1H),2.02-1.91(m,3H),1.90 -1.83(m,1H),1.71-1.66(m,3H).

[0291] Example 16. Preparation of Compound 16

[0292] Step 1: To a solution of compound 101a (500 mg, 4.95 mmol, Bid) in N,N-dimethylformamide (10 mL) at 0°C was added sodium hydride (396 mg, 9.89 mmol, 60% purity). The mixture was stirred at 0°C for 20 minutes. Compound 101b (1.73 mL, 7.42 mmol, Bid) was then added and stirred at 25°C for 18 hours. The mixture was poured into water (20 mL) and extracted with ethyl acetate (20 mL x 3). The combined organic phases were washed with saturated brine (20 mL x 4), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to yield the crude product. The crude product was purified by silica gel chromatography (eluent: petroleum ether-tetrahydrofuran = 20-40%) to yield compound 101c. MS m / z (ESI): 274.2 [M+1].

[0293] Step 2: Tetrabutylammonium fluoride (0.81 ml, 2.74 mmol) was added to a solution of compound 101c (0.50 g, 1.83 mmol) in tetrahydrofuran (5 mL) at 25°C, and the mixture was stirred at 25°C for 18 hours. The mixture was concentrated under reduced pressure to give the crude product, which was purified by silica gel chromatography (eluent: dichloromethane-methanol = 7-10%) to give compound 101d. MS m / z (ESI): 160.0 [M+1].

[0294] Step 3: To a solution of compound 101d (96 mg, 0.48 mmol) in tetrahydrofuran (4 mL) at 0°C was added sodium hydride (396 mg, 9.89 mmol, 60% purity). The reaction was stirred at 0°C for 20 minutes. Compound Int-2 (19 mg, 0.48 mmol) was then added and stirred at 25°C for 18 hours. The mixture was poured into water (10 mL) and extracted with ethyl acetate (10 mL x 3). The combined organic phases were washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to yield the crude product. The crude product was purified by HPLC using a Gilson 306-1741 column (Waters Xbridge C18-10 μm-19 x 250 mm; mobile phase: water (containing 0.1% ammonium bicarbonate) and acetonitrile, gradient: acetonitrile 0% to 95%, flow rate: 20 mL / min) to yield compound 16. MS m / z(ESI):537.0[M+1]. 1 H NMR (400MHz, DMSO-d6): δ10.01(s,1H),8.47(d,J=0.8Hz,1H),7.60(dd,J=8.6,2.6Hz,1H),7.47(s ,1H),6.95(d,J=8.2Hz,1H),4.76(d,J=11.8Hz,1H),4.56(d,J=11.8Hz,1H),4.45(t,J=6.4Hz,2H) ,4.14(t,J=5.4Hz,2H),3.69-3.54(m,2H),3.36-3.32(m,2H),3.29-3.26(m,2H),3.21-3.08(m,2H ),2.92-2.85(m,1H),2.71-2.67(m,1H),2.61-2.58(m,1H),2.26-2.22(m,5H),2.02-1.89(m,4H).

[0295] Example 17. Preparation of Compound 17

[0296] Step 1: At 0°C under N2 protection, compound 103a (1.00 g, 8.69 mmol, Bid) was dissolved in N,N-dimethylformamide (20 mL) and slowly added to a reaction flask containing sodium hydride (0.42 g, 60% purity, 10.42 mmol). After stirring at 0°C for 10 minutes, compound 103b (3.30 g, 13.03 mmol, Bid) was added and the mixture was heated to 25°C and stirred for 18 hours. The reaction mixture was poured into water (20 mL) and diluted, then extracted with ethyl acetate (30 mL x 3). The organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (eluent system: dichloromethane:methanol = 100:1 to 10:1) to obtain compound 103c. MS m / z (ESI): 288.1 [M+1].

[0297] Step 2: Dissolve compound 103c (1.00 g, 3.48 mmol) in tetrahydrofuran (15 mL), add tetrabutylammonium fluoride (5.22 mL, 5.22 mmol), and stir at 25°C for 2 hours. The reaction mixture was poured into water (10 mL), diluted, and extracted with ethyl acetate (20 mL x 3). The combined organic phases were washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (eluent system: dichloromethane:methanol = 100:1 to 10:1) to obtain compound 103d. MS m / z (ESI): 174.0 [M+1].

[0298] Step 3: At 0°C under N2 protection, compound 103d (50 mg, 0.29 mmol) was dissolved in tetrahydrofuran (2 mL) and slowly added to a reaction flask containing sodium hydride (11.60 mg, 60% purity, 0.48 mmol). After stirring at 0°C for 10 minutes, Int-2 (100 mg, 0.24 mmol) was added, and the mixture was heated to 25°C and stirred for 18 hours. The mixture was quenched with water (0.5 mL), and purified by HPLC using a Gilson 306-1741 column (Waters SunFire C18-10μm-19*250 mm; mobile phase: water (containing 0.1% ammonium bicarbonate) and acetonitrile, gradient: acetonitrile 0% to 95%, flow rate: 20 mL / min) to afford compound 17. MS m / z (ESI): 551.3 [M+1]. 1H NMR (400MHz, DMSO-d6): δ10.01(s,1H),8.47(s,1H),7.60(dd,J=8.6,2.6Hz,1H),7.47( d,J=2.6Hz,1H),6.95(d,J=8.8Hz,1H),4.76(d,J=11.8Hz,1H),4.56(d,J=11.8Hz,1H),4 .42(t,J=6.6Hz,2H),3.68–3.56(m,6H),3.49–3.39(m,4H),3.29(s,1H),3.20-3.09(m, 2H),2.89(s,1H),2.73–2.66(m,1H),2.61–2.58(m,2H),2.23(s,5H),1.94-1.88(m,2H).

[0299] Example 18. Preparation of Compound 18

[0300] Step 1: Compound 104a (195 mg, 0.86 mmol) was dissolved in acetonitrile (3 mL). Compound 104b (80 mg, 0.57 mmol, Bidex) and N,N-diisopropylethylamine (0.19 mL, 1.14 mmol, Bidex) were added, respectively, and stirred at 40°C for 1 hour. After completion of the reaction, the reaction solution was concentrated to dryness under reduced pressure, and the residue was purified by silica gel column chromatography (methanol:dichloromethane = 10%-18%) to obtain compound 104c. MS m / z (ESI): 333.2 [M+1].

[0301] Step 2: Compound 104c (150 mg, 0.45 mmol) was dissolved in a mixture of dichloromethane and acetonitrile (1:1, 4 mL) under ice. Sulfonyl chloride (364.70 μL, 4.51 mmol, Bidex) was added and stirred at 0°C for 3 hours. After completion of the reaction, the reaction solution was concentrated to dryness under reduced pressure, and the residue was purified by silica gel chromatography (methanol:dichloromethane = 30%-50%) to afford compound 104d. MS m / z (ESI): 320.9 [M+1].

[0302] Step 3: Compound 104e (500 mg, 2.01 mmol) was dissolved in dichloromethane (10 mL) at room temperature, followed by the addition of compound 104f (25.24 μL, 0.44 mmol, Bid) and stirring at 25°C for 1 hour. After the reaction, methanol (10 mL) was added to the reaction solution, which was then concentrated to dryness under reduced pressure. The residue was purified by silica gel column chromatography (methanol:dichloromethane = 1%-3%) to yield compound 104g. MS m / z (ESI): 292.0 [M+1].

[0303] Step 4: Compound 104g (200 mg, 0.69 mmol) was dissolved in tetrahydrofuran (5 mL), followed by the addition of wet palladium on carbon (73 mg, 0.07 mmol, 10%). The reaction mixture was purged with hydrogen three times and stirred at room temperature for 3 hours. After completion of the reaction, the reaction mixture was filtered, washed with ethyl acetate (30 mL), and the filtrate was concentrated to dryness to yield compound 104h. MS m / z (ESI): 262.1 [M+1].

[0304] Step 5: Compound 104h (100 mg, 0.38 mmol) was dissolved in isopropanol (2 mL), followed by the addition of compound 104d (98 mg, 0.30 mmol) and trifluoroacetic acid (436.32 mg, 3.83 mmol), respectively. The mixture was stirred at 40°C for 18 hours. After completion of the reaction, the reaction solution was purified by HPLC (Waters-Xbridge-C18-10μm-19*250mm column; mobile phase: water (containing 10 mM ammonium bicarbonate) and acetonitrile, gradient ratio: acetonitrile 30%-60%, flow rate: 25 mL / min) to afford compound 18. MS m / z (ESI): 546.3 [M+1]. 1 H NMR (400MHz, DMSO-d6): δ9.52(s,1H),8.29(s,1H),7.68-7.60(m,1H),7.58-7.52(m,1H),6 .93(d,J=8.4Hz,1H),4.80–4.65(m,1H),4.59–4.47(m,1H),4.02-3.41(m,9H),3.30-3.19( m,2H),3.15(t,J=8.0Hz,1H),2.91-2.77(m,1H),2.57(q,J=7.6Hz,1H),2.44(m,2H),2.35( d,J=9.2Hz,1H),2.23(s,3H),2.03(d,J=8.4Hz,3H),1.98-1.86(m,2H),1.81-1.69(m,2H).

[0305] Example 19. Preparation of Compound 19

[0306] Step 1: Compounds 105a (5.00 g, 23.12 mmol, Bid) and 105b (4.93 g, 23.12 mmol, Bid) were added to 1,4-dioxane (80 mL). N,N-diisopropylethylamine (11.49 mL, 4.01 mmol) was added at room temperature and stirred at 100°C for 18 hours. The reaction solution was concentrated to dryness, and the crude product was purified by silica gel column chromatography (eluent system: ethyl acetate:petroleum ether = 10:1 to 1:1) to obtain compound 105c. MS m / z (ESI): 364.4 [M+1].

[0307] Step 2: Compound 105c (2.40 g, 6.60 mmol) was added to tetrahydrofuran (30 mL). Lithium borohydride (0.33 mL, 0.66 mmol) and sodium borohydride (0.75 g, 19.81 mmol) were added separately at 0°C and allowed to react at room temperature for 18 hours. The reaction mixture was quenched with water (100 mL) and extracted with ethyl acetate (20 mL x 3). The combined organic phases were washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to dryness. The crude product was purified by silica gel column chromatography (eluent system: dichloromethane:methanol = 20:1 to 10:1) to obtain compound 105d. MS m / z (ESI): 368.4 [M+1].

[0308] Step 3: Compound 105d (4.00 g, 10.89 mmol) was added to 50% concentrated sulfuric acid (20 mL) and reacted at 120°C for 1 hour. The reaction solution was quenched with saturated sodium bicarbonate (1200 mL) and concentrated to dryness. The solid was washed with dichloromethane (50 mL x 3) and filtered. The filtrate was concentrated to dryness to provide compound 105e. MS m / z (ESI): 250.2 [M+1].

[0309] Step 4: Compound 105e (0.50 g, 2.01 mmol) and 105f (216.81 mg, 3.01 mmol, Bid) were added to 1,2-dichloroethane (10 mL). Sodium triacetoxyborohydride (0.85 g, 4.01 mmol) was added at room temperature and allowed to react for 18 hours. The reaction solution was quenched with saturated sodium bicarbonate (10 mL) and extracted with ethyl acetate (20 mL x 3). The combined organic phases were washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to yield compound 105g. MS m / z (ESI): 306.4 [M+1].

[0310] Step 5: Dissolve compound 105g (0.45g, 1.47mmol) in tetrahydrofuran (5mL) and add 10% palladium on carbon (0.16g, 0.15mmol). Stir the reaction mixture at room temperature under a hydrogen atmosphere for 18 hours. Filter the reaction mixture, wash the filter cake with tetrahydrofuran (5mL x 3), and concentrate the filtrate to dryness to obtain compound 105h. MS m / z (ESI): 276.4 [M+1].

[0311] Step 6: Compounds 105h (0.15 g, 0.54 mmol) and 105i (0.21 g, 0.65 mmol) were dissolved in isopropanol (5 mL). Trifluoroacetic acid (0.20 mL, 2.72 mmol) was added and the mixture was allowed to react at 40°C for 1 hour. The reaction solution was purified by HPLC (Gilson 306-1741, Shimadzu Shim-pack C18-5 μm-20 x 250 mm column; mobile phase: water (containing 10 mM ammonium bicarbonate) and acetonitrile, gradient: acetonitrile 30% to 95%, flow rate: 20 mL / min) to afford compound 19. MS m / z (ESI): 560.0 [M+1]. 1 H NMR (400MHz, DMSO-d6): δ9.51(s,1H),8.29(s,1H),7.62(s,1H),7.57(dd,J=8.8,2.6Hz,1H),6.91(d,J=8.6 Hz,1H),4.78(d,J=11.8Hz,1H),4.56(t,J=6.8Hz,3H),4.48–4.45(m,2H),3.63(dd,J=12.8,5.4Hz,3H),3.5 6(d,J=11.4Hz,2H),3.45(d,J=11.6Hz,2H),2.87(s,1H),2.72–2.63(m,2H),2.57(d,J=8.8Hz,2H),2.44(d, J=8.8Hz,2H),2.38–2.29(m,2H),2.23(s,3H),2.19–2.12(m,2H),1.93(dt,J=13.8,6.6Hz,2H),1.75(s,2H).

[0312] Example 20. Preparation of Compound 20

[0313] Compound Int-2 (0.10 g, 0.24 mmol) and 67a (30.50 mg, 0.24 mmol) were added to a solution of N,N-dimethylformamide (1 mL), followed by N,N-diisopropylethylamine (80.08 μL, 0.48 mmol). The mixture was stirred at 25°C for 1 hour. The reaction mixture was filtered, and the filtrate was purified by HPLC (column: Waters-Xbridge-C18-10 μm-19*250 mm; mobile phase: A: 10 mM ammonium bicarbonate / water; B: acetonitrile, gradient: 95% acetonitrile, flow rate: 20 mL / min) to afford compound 20. MS m / z (ESI): 504.4 [M+1]. 1 H NMR (400MHz, DMSO-d6): δ9.54(s,1H),8.23(s,1H),7.72–7.65(m,1H),7.56 –7.51(m,1H),6.94–6.88(m,1H),4.76(d,1H),4.52(d,1H),4.12(s,4H),3.6 8–3.54(m,2H),3.21–3.06(m,2H),2.90–2.82(m,1H),2.72–2.64(m,3H),2.6 1–2.55(m,1H),2.49–2.44(m,3H),2.23(d,J=7.4Hz,7H),2.12–2.04(m,2H).

[0314] Example 21. Preparation of Compound 21

[0315] Step 1: Compound 66b (0.83 g, 4.15 mmol, Bidex) and triethylamine (0.70 mL, 5.07 mmol) were dissolved in N,N-dimethylformamide (7 mL) and stirred at -10°C for 20 minutes. Compound 66a (0.62 mL, 4.61 mmol, Bidex) was then added and stirred at -10°C for 3 hours. The reaction mixture was diluted with water (30 mL) and extracted with ethyl acetate (20 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel column chromatography (eluent system: petroleum ether:ethyl acetate = 10:1 to 1:1) to yield compound 66c. MS m / z (ESI): 381.2 [M+1].

[0316] Step 2: Dissolve compound 66c (0.35 g, 0.92 mmol) in hydrochloric acid / ethyl acetate (7 mL, 4 M) and stir at room temperature for 2 hours. Concentrate the reaction mixture to dryness to obtain compound 66d. MS m / z (ESI): 281.2 [M+1].

[0317] Step 3: Compound 66d (0.32 g, 0.80 mmol) and N,N-diisopropylethylamine (0.66 mL, 3.99 mmol) were dissolved in N,N-dimethylformamide (5 mL). Compound 66e (66.23 mg, 0.56 mmol) was added dropwise at 0°C and stirred at 0°C for 30 minutes. The reaction mixture was diluted with water (20 mL) and extracted with ethyl acetate (20 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel column chromatography (eluent system: petroleum ether:tetrahydrofuran = 10:1 to 1:1) to obtain compound 66f. MS m / z (ESI): 363.2 [M+1].

[0318] Step 4: Compound 66f (86 mg, 0.24 mmol) and Int-1 (50 mg, 0.21 mmol) were dissolved in isopropanol (1 mL). Trifluoroacetic acid (0.08 mL, 1.07 mmol) was added, and the mixture was stirred at 40°C for 18 hours. The reaction mixture was purified by HPLC (Gilson 306-1741, Waters Xbridge C18-10 μm-19*250 mm column; mobile phase: water (containing 10 mmol NaHCO₃) and acetonitrile, gradient: acetonitrile 19% to 49%, flow rate: 25 mL / min) to afford compound 21. MS m / z (ESI): 560.4 [M+1]. 1 H NMR (400MHz, DMSO-d6): δ9.48(s,1H),8.16(s,1H),7.52(d,J=8.4Hz,2H),6.90(d,J=8.4Hz,1H),6.53(d,J= 7.6Hz,1H),4.77(d,J=11.6Hz,1H),4.53(d,J=11.8Hz,1H),4.47–4.25(m,2H),3.73(s,1H),3.68–3.53(m,2 H),3.35(dt,J=9.2,4.8Hz,2H),3.20–3.06(m,2H),2.91(dd,J=33.6,10.0Hz,2H),2.70(d,J=11.0Hz,1H),2 .63–2.54(m,2H),2.27–2.23(m,1H),2.22(s,3H),2.20–2.06(m,4H),1.92–1.72(m,4H),1.61–1.43(m,2H).

[0319] Example 22. Preparation of Compound 22

[0320] Compound Int-2 (0.13 g, 0.31 mmol) and 68a (56 mg, 0.38 mmol, Bid) were dissolved in N,N-dimethylformamide (2 mL). N,N-diisopropylethylamine (0.26 mL, 1.57 mmol) was added, and the mixture was stirred at 100°C for 1 hour. The reaction mixture was purified by HPLC (Gilson 306-1741, Waters Xbridge C18-10 μm-19*250 mm column; mobile phase: water (containing 10 mM NH4HCO3) and acetonitrile, gradient: acetonitrile 29% to 59%, flow rate: 25 mL / min) to afford compound 22. MS m / z (ESI): 490.2 [M+1]. 1 H NMR (400MHz, DMSO-d6): δ9.69–9.37(m,1H),8.22(s,1H),7.67(d,J=8.6Hz,1H),7.55(d,J= 2.6Hz,1H),6.91(d,J=8.6Hz,1H),4.76(d,J=11.8Hz,1H),4.53(d,J=11.8Hz,1H),4.25(s, 4H),3.68–3.55(m,2H),3.29(s,4H),3.20–3.09(m,2H),2.86(dd,J=8.2,4.0Hz,1H),2.70( d,J=11.0Hz,1H),2.62–2.56(m,1H),2.25(dd,J=7.6,3.4Hz,2H),2.22(s,3H),2.18(s,3H).

[0321] Example 23. Preparation of Compound 23

[0322] Compound 69a (36.6 mg, 0.29 mmol, Bid) was dissolved in tetrahydrofuran (2 mL), and sodium hydride (19.3 mg, 60% purity, 0.48 mmol) was added. After stirring at 0°C for 20 minutes, compound Int-2 (100 mg, 0.24 mmol) was added, and the mixture was stirred at 60°C for 18 hours. The reaction solution was purified by HPLC (Gilson 306-1741, column: Waters-Xbridge-C18-10μm-19*250mm; mobile phase: water (containing 10 mM NH4HCO3) and acetonitrile, gradient: acetonitrile 38%-68%, flow rate: 25 mL / min) to obtain compound 23. MS m / z (ESI): 504.4 [M+1]. 1H NMR (400MHz, DMSO-d6): δ10.04(s,1H),8.49(s,1H),7.68(d,J=2.2Hz,1H),7.57-7.54(m,1H),7.46(s ,1H),7.44(d,J=1.8Hz,1H),6.92(d,J=8.6Hz,1H),6.24-6.23(m,1H),4.75(d,J=11.8Hz,1H),4.54(d ,J=11.8Hz,1H),4.36(t,J=6.2Hz,2H),4.24(t,J=6.8Hz,2H),3.68–3.56(m,2H),3.21–3.08(m,2H),2 .91–2.86(m,1H),2.70(d,J=11.2Hz,1H),2.62–2.56(m,1H),2.28–2.24(m,2H),2.23(d,J=5.4Hz,5H).

[0323] Biological evaluation

[0324] Test Example 1: Compound Inhibition Experiment on ULK1 Protein Activity

[0325] 1. Test Purpose

[0326] In this experiment, the ADP-Glo ​​method was used to detect the inhibitory effect of the compound on the phosphorylation function of ULK1. 50 The inhibitory effects of the disclosed compounds on the ULK1 target were evaluated.

[0327] 2. Experimental Methods

[0328] Compound preparation: 3-fold serial dilutions were performed with DMSO using ECHO (Beckman, ECHO650) in a 384-well plate (PE, 6007290), with 10 concentration points. The highest concentration of the compound was 10 μM, and 50 nL was added to each well. 50 nL DMSO was added to the positive control wells and negative control wells, respectively.

[0329] Reaction procedure: Prepare a 32 nM ULK1 solution (Promega, V3521) and transfer 2.5 μL / well to the compound test wells and negative control wells (NC). Add 2.5 μL / well of 1× Assay buffer to the positive control wells (PC). Prepare a mixed solution of 100 μM ATP and 1.86 μM MBP and transfer 2.5 μL / well to the reaction plate. Initiate the reaction after centrifugation and incubate at room temperature for 60 minutes. After the reaction is complete, add 5 μL / well of ADP-Glo ​​Reagent (Promega, V9102) to the reaction plate, centrifuge, and incubate at room temperature for 40 minutes. Finally, add 10 μL / well of Kinase Detection Buffer; centrifuge and incubate at room temperature for 30 minutes. Chemiluminescence values ​​are read using Envision (PE, Envision2105).

[0330] Assay buffer: reaction buffer; Kinase Detection buffer: detection buffer, both are taken from the ADP-Glo ​​detection kit, purchased from Promega, model V9102.

[0331] 3. Data Analysis

[0332] The inhibition rate of the compound on ULK1 activity was calculated using the following formula:

[0333] Inhibition rate = (sample signal value - negative control signal value) / (positive control signal value - negative control signal value) x 100%

[0334] The log(inhibitor) vs. response-Variable slope of the analysis software GraphPad Prism 8 was used to fit the dose-effect curve to obtain the IC of each compound on the enzyme activity. 50 value.

[0335] The experimental results of this test case are shown in Table 1 below:

[0336] Table 1: IC values ​​of the compounds of the present invention for inhibition of ULK1 50

[0337] The compounds of the present invention have good inhibitory activity against ULK1. The activities of some compounds of the present invention measured by the above method are shown in Table 1, where A represents IC 50 <100nM; B means 100nM<IC 50 <1μM; C represents IC 50 >1μM.

[0338] Test Example 2: Inhibition of ATG13 phosphorylation levels in MIA PaCa-2 cells by compounds

[0339] 1. Test Purpose

[0340] The inhibitory effect of the disclosed compounds on ULK1 was evaluated by testing the inhibitory effect of the disclosed compounds on the phosphorylation level of ATG13 in MIA PaCa-2 cells.

[0341] 2. Experimental Methods

[0342] MIA PaCa-2 cells (ATCC, CRM-CRL-1420) were cultured in complete medium (DMEM (Gibco, 11965-092)) supplemented with 10% fetal bovine serum (Corning, 35-081-CV), 2.5% horse serum (Gibco, 16050-122), and 1% double-stranded antibody (Gibco, 15140-122). Experiments were performed according to the kit instructions (Cell Signaling Technology, 17208C).

[0343] Cell plating: MIA PaCa-2 cells with a confluence of 80% were digested, diluted with complete medium, and added to a 96-well cell culture plate (Costar, 3596), 20,000 cells per well, 90 μL.

[0344] Compound Incubation: Add 5 μL of Torin1 and 5 μL of serially diluted compound to each well to achieve a maximum compound concentration of 10 μM. Perform a five-fold serial dilution across eight concentrations, with a final Torin1 concentration of 100 nM. The DMSO content in the system is 0.1%. Incubate the reaction plate in an incubator at 37°C, 5% CO2 for 24 hours. Add 10 μL of Torin1 to the negative control well (NC) to achieve a final concentration of 100 nM. The DMSO content is 0.1%.

[0345] Reaction Assay: After incubation, wash the cell plate with pre-chilled DPBS (Corning, 21-031-CVC) to remove the compound and culture medium. Add 50 μL of pre-chilled cell lysis buffer to each well and incubate on ice for 10 minutes. Centrifuge the cell plate and transfer the supernatant to an ELISA plate. Simultaneously, add 50 μL of Phospho-Atg13 (ser355) rabbit detection antibody and incubate at room temperature with shaking for 60 minutes. After incubation, wash the plate four times with washing buffer and add 100 μL of TMB substrate for color development. Incubate in the dark with shaking for 15 minutes. Finally, add 100 μL of stop solution to each well and shake for 10 seconds to terminate the reaction. Over 10 minutes, absorbance measurements at 450 nm and 560 nm are collected using an Envision (Perkin Elmer, EnVision 2105).

[0346] 3. Data Analysis

[0347] The inhibition rate was calculated by subtracting the absorbance at 560 nM from the absorbance at 450 nm for each well. The inhibition rate formula is as follows:

[0348] Inhibition rate = (sample signal value - negative control signal value) / (positive control signal value - negative control signal value) x 100%

[0349] The dose-effect curve was fitted using the log(inhibitor) vs. response-Variable slope of Graphpad Prism8 to obtain the IC of each compound for the inhibition of ATG13 phosphorylation level. 50 value.

[0350] The experimental results of this test case are shown in Table 2 below:

[0351] Table 2: IC values ​​of the compounds of the present invention for inhibiting ATG13 phosphorylation levels 50

[0352] The compounds of the present invention have good inhibitory activity on the phosphorylation level of ATG13. The activities of some compounds of the present invention measured by the above method are shown in Table 2, where A represents IC 50 <100nM; B means 100nM<IC 50 <1μM; C represents IC 50 >1μM.

[0353] Test Example 3: Inhibition of autophagy levels in HEK293 GFP-LC3 stably transfected cells by compounds

[0354] 1. Test Purpose

[0355] The inhibitory effect of the disclosed compounds on intracellular autophagy was evaluated by testing the effect of the disclosed compounds on the fluorescence intensity of HEK293 GFP-LC3 stably transfected cells.

[0356] 2. Experimental Methods

[0357] HEK293 GFP-LC3 stably transfected cells (HEK293, ATCC, CRL-1573) were cultured in complete medium, DMEM (Gibco, 11965-092) containing 10% fetal bovine serum (Corning, 35-081-CV), 1% anti-antibody (Gibco, 15140-122), and 1 μg / mL Puromycin (Invitrogen-A1113803).

[0358] Cell plating: HEK293 GFP-LC3 stably transfected cells with a confluence of 80% were digested, diluted with culture medium without Puromycin, and added to 96-well cell culture plates (Geriner, 655090), with 12,000 cells per well in 90 μL.

[0359] Compound Incubation: Add 5 μL of Torin1 and 5 μL of serially diluted compound to each well, achieving a maximum compound concentration of 10 μM. Serially dilute the compound three-fold over eight concentrations, for a final Torin1 concentration of 100 nM. The DMSO content in the system is 0.1%. Positive control wells (PC) contain a final concentration of 0.1% DMSO, and negative control wells (NC) contain 100 nM Torin1 in 0.1% DMSO. Incubate the reaction plate in an incubator at 37°C, 5% CO2 for 24 hours.

[0360] Cell staining and fixation: Remove the cell plate and add 10 μL of Hochest 33342 nuclear staining reagent (Thermo, H1399) per well. Incubate at 37°C, 5% CO2 for 30 min. After incubation, discard the cell supernatant and add 100 μL of pre-chilled 4% paraformaldehyde per well to fix the cells for 15 min. The fixative solution is discarded, and 100 μL of PBS is added per well. The cells are then imaged and analyzed using a high-content cell imaging analyzer (PE, Operetta CLS). GFP fluorescence signals are collected for data processing.

[0361] 3. Data Analysis

[0362] The average GFP fluorescence signal intensity in the reaction wells was collected and the inhibition rate was calculated: Inhibition rate = (sample signal value - negative control signal value) / (positive control signal value - negative control signal value) x 100%

[0363] The log(inhibitor) vs. response-Variable slope of Graphpad Prism8 was used to fit the dose-effect curve to obtain the IC of each compound on the inhibition of autophagy level in HEK293 GFP-LC3 stably transfected cells. 50 value.

[0364] The experimental results of this test case are shown in Table 3 below:

[0365] Table 3: IC values ​​of the compounds of the present invention for inhibiting autophagy levels in HEK293 GFP-LC3 stably transfected cells 50

[0366] The compounds of the present invention have good inhibitory activity on the autophagy level in HEK293 GFP-LC3 stably transfected cells. The activities of some compounds of the present invention measured by the above method are shown in Table 3, where A represents IC 50 <100nM; B means 100nM<IC 50 <1μM; C represents IC 50 >1μM.

Claims

1. A compound of Formula 1 or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, hydrate, N-oxide, isotope-labeled compound, metabolite, ester, or prodrug thereof: in, X1, X2, and X3 are independently CH or N; Ring A is selected from 3-10 membered heterocyclic group, C3-C 10 Cycloalkyl, C6-C 10 Aryl and 5-10 membered heteroaryl; Ring B is selected from 3-10 membered heterocyclic group, C3-C 10 Cycloalkyl, C6-C 10 Aryl and 5-10 membered heteroaryl; R1 is independently selected from H, halogen, cyano, nitro, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, C3-C 10 Cycloalkyl, 3-10 membered heterocyclic group, C6-C 10 Aryl and 5-10 membered heteroaryl; R2 is selected from H, cyano, halogen, C1-C6 alkyl, C1-C6 hydroxyalkyl, C1-C6 aminoalkyl, C1-C6 haloalkyl, C1-C6 heteroalkyl, C2-C6 alkenyl, C2-C6 alkynyl and C3-C 10 Cycloalkyl; R3 and R4 are each independently selected from H, oxo, halogen, cyano, nitro, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C 10 Cycloalkyl, 3-10 membered heterocyclic group, C6-C 10 aryl, 5-10 membered heteroaryl, R5O-, R6C(O)-, -NR7R8 and -NHCONR7R8, wherein the C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C 10 Cycloalkyl, 3-10 membered heterocyclic group, C6-C 10 Aryl or 5-10 membered heteroaryl are each optionally substituted with one or more R9; R5 and R6 are each independently selected from H, C1-C6 alkyl, C3-C 10 Cycloalkyl, C6-C 10 Aryl and 5-10 membered heteroaryl; R7 and R8 are each independently selected from H, C1-C6 alkyl, C3-C 10 Cycloalkyl, 3-10 membered heterocyclic group and C6-C 10 Aryl; or R7, R8 and the N atom to which they are attached together form a 5-6 membered heterocyclic ring or a 5-6 membered heteroaromatic ring; R9 is independently selected from halogen, cyano, nitro, amino, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C 10 Cycloalkyl, 3-10 membered heterocyclic group, C6-C 10 Aryl and 5-10 membered heteroaryl; R is selected from -NR7R8, R5O-, R5S-, halogen, C1-C6 alkyl, 3-12 membered heterocyclyl, 5-10 membered heteroaryl and -LR 10 wherein the 3-12 membered heterocyclyl or 5-10 membered heteroaryl are each optionally substituted by one or more R 11 replace; L is -(L1) m -(L2) n -(L3) p -; L1 is selected from -O-, -N(R 12 )-、-C(R 12 )(R 13 )-O-、-N(R 12 )CO- and -S-; L2 is selected from C1-C6 alkylene, C1-C6 heteroalkylene, C3-C 10 Cycloalkylene and 3-10 membered heterocyclylene; L3 is selected from -N(R 12 )-、-N(R 12 )C(O)- and -C(O)-; m, n and p are each independently selected from 0, 1 and 2; R 10 Selected from C3-C 10 Cycloalkyl, C6-C 10 aryl, 5-10 membered heteroaryl and 3-10 membered heterocyclic group, the C3-C 10 Cycloalkyl, C6-C 10 Aryl, 5-10 membered heteroaryl or 3-10 membered heterocyclyl are each optionally substituted by one or more R 14 replace; R 11 Each is independently selected from C1-C6 alkyl, oxo, 5-10 membered heteroaryl, 3-10 membered heterocyclyl, R6C(O)-, R6C(O)N(R 12 )- and -NR7R8, wherein the C1-C6 alkyl, 5-10 membered heteroaryl or 3-10 membered heterocyclyl is each optionally substituted by one or more R 15 replace; R 12 , R 13 Each independently selected from H, C1-C6 alkyl and C3-C 10 Cycloalkyl; R 14 are each independently selected from H, halogen, cyano, oxo, C1-C6 alkyl, C1-C6 alkoxy, -CONR7R8 and R6C(O)-; and R 15 Each is independently selected from H, halogen, cyano, oxo, C1-C6 alkyl, C3-C 10 Cycloalkyl and 3-10 membered heterocyclic groups, wherein the C1-C6 alkyl, C3-C 10 The cycloalkyl or 3-10 membered heterocyclyl are each optionally substituted with one or more substituents selected from C1-C6 alkyl and oxo; and x, y, z are each independently selected from 1, 2 and 3.

2. The compound of claim 1 or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, hydrate, N-oxide, isotope-labeled compound, metabolite, ester, or prodrug thereof, wherein: At most one of X1, X2, and X3 is N.

3. The compound of claim 1 or 2, or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, hydrate, N-oxide, isotope-labeled compound, metabolite, ester, or prodrug thereof, wherein: Ring A is a 5-8 membered heterocyclic group, preferably a 6-7 membered heterocyclic group, more preferably a 6-7 membered nitrogen-containing heterocyclic group; Preferably, ring A is in, X4 is selected from O and NH; d is 0 or 1; e is 1 or 2; Wavy Lines It indicates the point of attachment of the group to the rest of the molecule; Symbol 1 represents ring A and The connection point of the part; and Symbol 2 represents the connection point between ring A and ring B; Preferably, ring A is selected from 4. The compound of any one of claims 1 to 3 or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, hydrate, N-oxide, isotope-labeled compound, metabolite, ester, or prodrug thereof, wherein: Ring B is a 5-6 membered heterocyclic ring, more preferably a 5-6 membered nitrogen-containing heterocyclic ring; Preferably, ring B is The wavy line Indicates the point of attachment of ring B to ring A.

5. The compound of any one of claims 1 to 4 or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, hydrate, N-oxide, isotope-labeled compound, metabolite, ester, or prodrug thereof, wherein: R1 is H.

6. The compound of any one of claims 1 to 5, or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, hydrate, N-oxide, isotope-labeled compound, metabolite, ester, or prodrug thereof, wherein: R2 is selected from H and C1-C6 haloalkyl, preferably C1-C6 haloalkyl, further preferably C1-C3 haloalkyl, and more preferably trifluoromethyl.

7. The compound of any one of claims 1 to 6 or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, hydrate, N-oxide, isotope-labeled compound, metabolite, ester, or prodrug thereof, wherein: R3 and R4 are each independently selected from H, oxo, C1-C6 alkyl, C3-C 10 Cycloalkyl, 3-10 membered heterocyclic group and R6C(O)-, wherein the C1-C6 alkyl, C3-C 10 Cycloalkyl or 3-10 membered heterocyclyl are each optionally substituted with one or more R9; R6 is selected from C1-C6 alkyl and C3-C 10 cycloalkyl; and R9 are each independently selected from halogen and C1-C6 alkyl; Preferably, R3, R4 are each independently selected from H, oxo, C1-C6 alkyl, C3-C6 cycloalkyl, 4-6 membered heterocyclyl and R6C(O)-, wherein the C1-C6 alkyl is optionally substituted by 1, 2, 3, 4 or 5 halogens, and the 4-6 membered heterocyclyl is optionally substituted by 1, 2 or 3 C1-C6 alkyl; and R6 is a C1-C6 alkyl group; More preferably, R3, R4 are each independently selected from H, oxo, C1-C6 alkyl, C3-C6 cycloalkyl, oxetanyl, piperidinyl, tetrahydropyranyl and R6C(O)-, wherein the C1-C6 alkyl is optionally substituted with 1, 2, 3, 4 or 5 halogens, and the piperidinyl or tetrahydropyranyl are each optionally substituted with 1 or 2 C1-C6 alkyls; and R6 is a C1-C6 alkyl group; More preferably, R3 and R4 are each independently selected from H, oxo, methyl, isopropyl, cyclopropyl, cyclobutyl, CF3CH2-, CH3CO-, The wavy line Indicates the point of attachment of a group to the rest of the molecule.

8. The compound of claim 7 or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, hydrate, N-oxide, isotope-labeled compound, metabolite, ester, or prodrug thereof, wherein: R3 is selected from H, oxo and C1-C6 alkyl, preferably selected from H, oxo and C1-C3 alkyl, more preferably selected from H, oxo and methyl.

9. The compound of claim 7 or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, hydrate, N-oxide, isotope-labeled compound, metabolite, ester, or prodrug thereof, wherein: R4 is selected from C1-C6 alkyl, C3-C6 cycloalkyl, 4-6 membered heterocyclyl and R6C(O)-, wherein the C1-C6 alkyl is optionally substituted by 1, 2, 3, 4 or 5 halogens, and the 4-6 membered heterocyclyl is optionally substituted by 1, 2 or 3 C1-C6 alkyl; and R6 is a C1-C6 alkyl group; Preferably, R4 is selected from C1-C6 alkyl, C3-C6 cycloalkyl, oxetanyl, piperidinyl, tetrahydropyranyl and R6C(O)-, wherein the C1-C6 alkyl is optionally substituted by 1, 2, 3, 4 or 5 halogens, and the piperidinyl or tetrahydropyranyl are each optionally substituted by 1 or 2 C1-C6 alkyl; and R6 is a C1-C6 alkyl group; More preferably, R4 is selected from methyl, isopropyl, cyclopropyl, cyclobutyl, CF3CH2-, CH3CO-, The wavy line Indicates the point of attachment of a group to the rest of the molecule.

10. The compound of any one of claims 1 to 9 or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, hydrate, N-oxide, isotope-labeled compound, metabolite, ester, or prodrug thereof, wherein: x, y, z are each independently selected from 1 and 2.

11. The compound of any one of claims 1 to 10 or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, hydrate, N-oxide, isotope-labeled compound, metabolite, ester, or prodrug thereof, wherein: R is selected from 3-12 membered heterocyclyl, 5-10 membered heteroaryl and -LR 10 wherein the 3-12 membered heterocyclyl or 5-10 membered heteroaryl are each optionally substituted by one or more R 11 replace; L is -(L1) m -(L2) n -(L3) p -; L1 is selected from -O-, -NH- and -CH2-O-; L2 is selected from C1-C6 alkylene, C3-C 10 Cycloalkylene and 3-10 membered heterocyclylene; L3 is selected from -NH-, -NHC(O)- and -C(O)-; m, n and p are each independently selected from 0 and 1, and m, n and p are not 0 at the same time; R 10 Selected from C3-C 10 Cycloalkyl, C6-C 10 aryl, 5-10 membered heteroaryl and 3-10 membered heterocyclic group, the C3-C 10 Cycloalkyl, C6-C 10 Aryl, 5-10 membered heteroaryl or 3-10 membered heterocyclyl are each optionally substituted by one or more R 14 replace; R 11 Each is independently selected from C1-C6 alkyl, oxo, 5-10 membered heteroaryl, 3-10 membered heterocyclyl and R6C(O)-, wherein the 3-10 membered heterocyclyl is each optionally substituted by one or more R 15 replace; R 14 are each independently selected from H, halogen, cyano, oxo and C1-C6 alkyl; and R 15 Each is independently selected from oxo and C1-C6 alkyl.

12. The compound of any one of claims 1 to 11 or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, hydrate, N-oxide, isotope-labeled compound, metabolite, ester, or prodrug thereof, wherein: L is -(L1) m -(L2) n -(L3) p -*, where the bond marked with "*" is connected to R 10 ; L1 is selected from -O-, -N(R 12 )-、-C(R 12 )(R 13 )-O-@、-N(R 12 )CO-@ and -S-, wherein the bond marked with "@" is connected to L2; L2 is selected from C1-C6 alkylene, C1-C6 heteroalkylene, C3-C 10 Cycloalkylene and 3-10 membered heterocyclylene; L3 is selected from -N(R 12 )-、-N(R 12 )C(O)-$ and -C(O)-, where the bond marked with "$" is connected to R 10 ; m, n and p are each independently selected from 0 and 1, and m, n and p are not 0 at the same time; Preferably, L1 is selected from -O-, -N(R 12 )-; L2 is selected from C1-C6 alkylene, C3-C6 cycloalkylene and 3-6 membered heterocyclylene; L3 is selected from -N(R 12 )-、-N(R 12 )C(O)-$ and -C(O)-, more preferably -C(O)-, wherein the bond marked with "$" is connected to R 10 ; Preferably, L1 is selected from -O-, -NH-; L2 is selected from ethylene, propylene, cyclohexylene, piperidinylene; L3 is selected from -NH-, -NH-C(O)-$, -C(O)-, wherein the bond marked with "$" is connected to R 10 ; Preferably, L is -O-, -O-cyclohexylene-*, -NH-cyclohexylene-*, -NH-piperidinylene-C(O)-*, -O-propylene-*, -NH-propylene-*, -O-ethylene-NH-*, -ethylene-NHC(O)-*, -NH-propylene-C(O)-*, and -O-propylene-C(O)-*, wherein the bond marked with "*" is connected to R 10 ; More preferably, L is selected from -O-propylene-*, -O-cyclohexylene-*, -NH-propylene-*, -NH-cyclohexylene-*, -NH-piperidinylene-C(O)-*, wherein the bond marked with "*" is connected to R 10 .

13. The compound of any one of claims 1 to 12, or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, hydrate, N-oxide, isotopically labeled compound, metabolite, ester, or prodrug thereof, wherein the compound is a compound of formula 1':

14. The compound of any one of claims 1 to 13, or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, hydrate, N-oxide, isotopically labeled compound, metabolite, ester, or prodrug thereof, wherein the compound is a compound of Formula 1A, 1B, IC or ID:

15. The compound of claim 14 or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, hydrate, N-oxide, isotopically labeled compound, metabolite, ester, or prodrug thereof, wherein the compound is a compound of Formula 1A-1: in, Ring C is selected from 3-12 membered heterocyclyl and 5-10 membered heteroaryl; preferably, Ring C is selected from 3-12 membered heterocyclyl; and r is selected from 0, 1 and 2.

16. The compound of claim 15 or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, hydrate, N-oxide, isotope-labeled compound, metabolite, ester, or prodrug thereof, wherein: Ring C is a 5-12 membered spiro heterocyclic group or a 5-12 membered bridged heterocyclic group, preferably a 7-12 membered spiro heterocyclic group or a 7-12 membered bridged heterocyclic group, more preferably More preferably and R 11 Each independently selected from C1-C6 alkyl and oxo, preferably, R 11 are each independently selected from methyl and oxo, The wavy line Indicates the point of attachment of a group to the rest of the molecule.

17. The compound of claim 15 or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, hydrate, N-oxide, isotope-labeled compound, metabolite, ester, or prodrug thereof, wherein: Ring C is a 3-10-membered monocyclic heterocyclic group or a 5-10-membered heteroaryl group, preferably a 6-7-membered monocyclic heterocyclic group or a 5-6-membered heteroaryl group, more preferably a piperidinyl group, a piperazinyl group, an azepanyl group, a 1,4-diazepanyl group or a pyrazolyl group; R 11 Each is independently selected from C1-C6 alkyl, oxo, 5-6 membered heteroaryl, 5-7 membered heterocyclyl and R6C(O)-, wherein the C1-C6 alkyl, 5-6 membered heteroaryl or 5-7 membered heterocyclyl is each optionally substituted by 1, 2, 3 or 4 R 15 replace; R6 is C4-C6 cycloalkyl; and R 15 are each independently selected from oxo and C1-C6 alkyl, Preferably, ring C is a 3-10 membered monocyclic heterocyclic group, preferably a 6-7 membered monocyclic heterocyclic group, more preferably a piperidinyl group, a piperazinyl group, an azepanyl group, or a 1,4-diazepanyl group; Preferably, R 11 Each is independently selected from C1-C6 alkyl, oxo, pyrazolyl, piperidinyl and C4-C6 cycloalkyl-C(O)-, wherein the piperidinyl is optionally substituted with 1 or 2 substituents selected from C1-C6 alkyl and oxo; More preferably, R 11 Each is independently selected from methyl, oxo, pyrazolyl, N-methylpiperidinyl, oxopiperidinyl and cyclobutylcarbonyl.

18. The compound of any one of claims 15-17, or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, hydrate, N-oxide, isotopically labeled compound, metabolite, ester, or prodrug thereof, wherein the compound is a compound of Formula 1A-1a, 1A-1b, or 1A-1c: in, t is selected from 0, 1, 2 and 3; u, v, w, a, b and c are each independently selected from 1, 2 and 3; Y is selected from -CH- and -N-; and r is selected from 1 and 2.

19. The compound of any one of claims 1 to 14, or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, hydrate, N-oxide, isotopically labeled compound, metabolite, ester, or prodrug thereof, wherein the compound is a compound of Formula 1A-2: in, L is -(L1) m -(L2) n -(L3) p -, preferably, L is -(L1) m -(L2) n -(L3) p -*, wherein the bond marked with "*" is connected to ring D; L1 is selected from -O-, -NH- and -CH2-O-. Preferably, when L1 is -CH2-O-, the oxygen of -CH2-O- is connected to L2. Preferably, L1 is selected from -O-, -NH-; L2 is selected from C1-C6 alkylene, C3-C 10 Cycloalkylene and 3-10 membered heterocyclylene, preferably selected from C1-C6 alkylene, C3-C6 cycloalkylene and 3-6 membered heterocyclylene, preferably selected from C1-C6 alkylene and C3-C6 cycloalkylene, preferably selected from ethylene, propylene, cyclohexylene and piperidinylene, more preferably selected from ethylene, propylene and cyclohexylene; L3 is selected from -NH-, -NHC(O)- and -C(O)-. Preferably, when L3 is -NHC(O)-, the amino portion of the -NHC(O)- is connected to L2, and the carbonyl portion is connected to ring D; m, n and p are each independently selected from 0 and 1, and m, n and p are not 0 at the same time; Preferably, L is selected from -O-, -O-cyclohexylene-, -NH-cyclohexylene-, -NH-piperidinylene-, -O-propylene-, -NH-propylene-, -ethylene-, -ethylene-NHC(O)-, and -O-propylene-C(O)-; Preferably, L is selected from -O-, -O-cyclohexylene-, -NH-cyclohexylene-, -NH-piperidinylene-C(O)-, -O-propylene-, -NH-propylene-, -O-ethylene-NH-, -ethylene-NHC(O)-, -NH-propylene-C(O)-, and -O-propylene-C(O)-; Preferably, L is selected from -O-, -O-cyclohexylene-*, -NH-cyclohexylene-*, -NH-piperidinylene-C(O)-*, -O-propylene-*, -NH-propylene-*, -O-ethylene-NH-*, -ethylene-NHC(O)-*, -NH-propylene-C(O)-*, and -O-propylene-C(O)-*, wherein the bond marked with "*" is connected to ring D; More preferably, L is selected from -O-propylene-*, -O-cyclohexylene-*, -NH-propylene-*, -NH-cyclohexylene-*, -NH-piperidinylene-C(O)-*, wherein the bond marked with "*" is connected to ring D; Ring D is selected from C3-C 10 Cycloalkyl, C6-C 10 Aryl, 5-10 membered heteroaryl and 3-10 membered heterocyclic group, preferably selected from C3-C6 cycloalkyl, C6-C 10 Aryl, 5-6 membered heteroaryl and 5-7 membered heterocyclyl, preferably selected from cyclobutyl, phenyl, pyrazolyl, piperidinyl, piperazinyl, pyrrolidinyl, imidazolidinyl, morpholinyl, 1,3-oxazinyl, azepanyl and 1,4-oxaazepanyl; R 14 are each independently selected from H, halogen, cyano, oxo and C1-C6 alkyl; and s is selected from 0, 1 and 2.

20. The compound of claim 19, or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, hydrate, N-oxide, isotopically labeled compound, metabolite, ester, or prodrug thereof, wherein the compound is a compound of Formula 1A-2a, 1A-2b, 1A-2c, 1A-2d, 1A-2e, or 1A-2f: in, X is selected from -O-, -NH- and -CH2-O-, preferably, X is selected from -O- and -NH-; V is selected from -O-, -S-, -NR 12 -and-CHR 12 -, preferably, V is selected from -O- and -CHR 12 -; R 12 Selected from H and C1-C6 alkyl; h and l are each independently selected from 0, 1, 2 and 3; i is selected from 0, 1 and 2; and j and k are each independently selected from 1, 2 and 3.

21. The compound of claim 14 or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, hydrate, N-oxide, isotopically labeled compound, metabolite, ester, or prodrug thereof, wherein the compound is a compound of Formula 1B-2a, 1C-2a, or 1D-2a: in, X is selected from -O-, -NH- and -CH2-O-, preferably, X is selected from -O- and -NH-, more preferably, X is selected from -O-; V is selected from -O-, -S-, -NR 12 -and-CHR 12 -, preferably, V is selected from -CHR 12 -; R 12 Selected from H and C1-C6 alkyl; h and l are each independently selected from 0, 1, 2 and 3; and i is selected from 0, 1 and 2.

22. The compound of claim 1 or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, hydrate, N-oxide, isotopically labeled compound, metabolite, ester, or prodrug thereof, wherein the compound is selected from:

23. A pharmaceutical composition comprising an effective amount of a compound of any one of claims 1 to 22 or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, hydrate, N-oxide, isotopically labeled compound, metabolite, ester, or prodrug thereof, and one or more pharmaceutically acceptable carriers.

24. Use of a compound according to any one of claims 1 to 22 or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, hydrate, N-oxide, isotopically labeled compound, metabolite, ester, or prodrug thereof, or a pharmaceutical composition according to claim 23, in the preparation of a medicament for treating a ULK1-mediated disease in a subject in need thereof; preferably, wherein the ULK1-mediated disease is characterized by abnormal autophagy; preferably, wherein the abnormal autophagy is induced by treatment; preferably, wherein the ULK1-mediated disease is cancer, preferably lung cancer, breast cancer or pancreatic cancer, such as non-small cell lung cancer, triple-negative breast cancer or pancreatic ductal adenocarcinoma, or the ULK1-mediated disease is tuberous sclerosis complex (TSC) or lymphangioleiomyomatosis (LAM).

25. The use according to claim 24, wherein the medicament is for co-administration with an additional therapeutic agent; preferably, wherein the additional therapeutic agent is a standard of care therapy, such as an mTOR inhibitor, carboplatin, a MEK inhibitor or a PARP inhibitor.

26. The use according to claim 24 or 25, wherein the medicament is used to degrade ATG13 in the subject.

27. A method for preparing the compound of claim 1, comprising the following route A or route B: in, R1, R2, R3, R4, x, y, z, X1, X2 and X3 are as defined in claim 1.

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