Quinazoline compound as well as preparation method and application thereof

Abstract

The invention relates to a quinazoline compound as well as a preparation method and application thereof. Specifically, the invention relates to a compound as shown in a formula (I) or pharmaceutically acceptable salt, stereoisomer, tautomer, polymorphic substance, solvate, N-oxide, isotope labeled compound, metabolite or prodrug thereof, and a pharmaceutical composition containing the compound or the pharmaceutically acceptable salt, the stereoisomer, the tautomer, the polymorphic substance, the solvate, the N-oxide, the isotope labeled compound, the metabolite or the prodrug thereof. The invention also relates to a preparation method of the compound and application of the compound in preparation of drugs for preventing or treating KRAS G12D-mediated related diseases. # imgabs0 #

Description

Technical Field

[0001] The present invention belongs to the field of medicine, and in particular relates to a quinazoline compound, a preparation method thereof and a use thereof. Background Art

[0002] RAS is a guanine nucleotide-binding protein with GTPase activity. It can switch between the GDP-bound inactive state and the GTP-bound active state, affecting multiple downstream signaling pathways such as Raf, PI3K and RalGDS, and regulating protein synthesis, gene transcription, cell growth, differentiation, apoptosis and migration.

[0003] RAS mutations lead to persistent activation of downstream signaling pathways, promoting tumor development and progression. Across all tumor types, RAS mutations primarily occur in KRAS (85%), with the most frequent mutation occurring at position 12 of KRAS, glycine to aspartic acid (G12D), at 34.2%. In KRAS-mutated tumor cells, KRAS GTPase activity decreases, remaining persistently active. KRAS mutations are closely associated with the development of lung, pancreatic, and colorectal cancers.

[0004] The development of KRAS G12D degraders is still in the clinical validation stage. WO2020173935 discloses a class of RAS inhibitors that utilize molecular glue principles to induce KRAS G12D to form dimers, thereby blocking the interaction between KRAS and downstream proteins.

[0005] Proteolysis-Targeting Chimeras (PROTAC) are bifunctional molecules whose structure consists of three parts: (1) a part that binds to the target protein substrate; (2) a part that binds to the E3 ubiquitin ligase; and (3) a chain connecting the first two parts. PROTAC can recognize the target protein and E3 ubiquitin ligase respectively, shorten the distance between the target protein and the E3 ubiquitin ligase, and induce the ubiquitination process and induce the degradation of the target protein by inducing the recruitment of the E3 ubiquitin ligase to the surface of the target protein. PROTAC has the advantages of a wide range of pharmacological activities, high target selectivity, can be used to degrade difficult-to-drug target proteins, has strong degradation efficacy, and can maintain catalytic degradation at low concentrations.

[0006] There is an urgent need in this field to develop PROTACs targeting KRAS G12D protein with novel structure and good biological activity. SUMMARY OF THE INVENTION

[0008] The present invention provides degraders that can be used to target the KRAS G12D protein. The compounds of the present invention are PROTACs, one end of which can bind to the target protein KRAS G12D, while the high-affinity VHL ligand at the other end can recruit the target protein to the E3 ubiquitin ligase, leading to ubiquitination and subsequent degradation of the KRAS G12D protein. The compounds of the present invention can be used to treat and / or prevent cancers or diseases caused by aggregation and / or overactivation of the target protein.

[0009] In one aspect, the present invention provides a compound or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite or prodrug thereof, wherein the compound has the structure of Formula (I):

[0010]

[0011] in:

[0012] Ring A is a 3-7 membered heterocyclic group, and Ring A is optionally substituted by one or more R 7 replace;

[0013] Ring E is a 6-12 membered spiro heterocycle, a 6-12 membered heterocycle or a 6-12 membered bridged heterocycle, and is optionally substituted by one or more R 8 replace;

[0014] Ring X is a benzene ring or a 5-6 membered heteroaromatic ring, and ring X is optionally substituted by one or more R 9 replace;

[0015] Ring Y is a 5-6 membered heteroaromatic ring;

[0016] Ring Z is a benzene ring or a 5-6 membered heteroaromatic ring, and is optionally substituted by one or more R 10 replace;

[0017] L 1 is a covalent bond, or is selected from O, S and NR 11 ;

[0018] L 2 is a covalent bond, or is selected from O, S and NR 12 ;

[0019] L 3 Selected from C 1-6 Alkylene, -OC 1-6 Alkylene-, -C 1-6 Alkylene-O- and -C 1-6 Alkylene-OC 1-6 Alkylene-;

[0020] R 1 Selected from hydrogen, halogen, cyano, C 1-6Alkyl, C 2-6 Alkenyl, C 3-6 Cycloalkyl, C 1-6 Haloalkyl, -OC 1-6 Alkyl, -OC 1-6 Haloalkyl and -OC 3-6 Cycloalkyl;

[0021] R 2 Selected from hydrogen, halogen, cyano, C 1-6 Alkyl, C 2-6 Alkenyl, C 3-6 Cycloalkyl, C 1-6 Haloalkyl, -OC 1-6 Alkyl and -OC 1-6 alkyl halide;

[0022] Each R 3 independently selected from hydrogen, halogen, cyano, C 1-6 Alkyl, C 1-6 Haloalkyl and C 3-6 Cycloalkyl;

[0023] R 4 Selected from C 1-6 Alkyl, C 1-6 Haloalkyl and C 3-6 Cycloalkyl;

[0024] R 5a and R 5b are each independently selected from hydrogen, C 1-6 Alkyl, C 1-6 Haloalkyl, -C 1-6 Alkylene -OH and C 3-6 Cycloalkyl; or

[0025] R 5a and R 5b Together with the carbon atoms connected to form C 3-6 Cycloalkyl or 3-6 membered heterocyclic group;

[0026] Each R 6 independently selected from hydrogen, halogen, cyano, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 3-6 Cycloalkyl, -OC 1-6 Alkyl and -OC 1-6 alkyl halide;

[0027] R 7 Selected from hydrogen, C 1-6 Alkyl, C 1-6 Haloalkyl and C 3-6 Cycloalkyl;

[0028] R8 Selected from hydrogen, halogen, C 1-6 Alkyl, C 1-6 Haloalkyl and C 3-6 Cycloalkyl;

[0029] R 9 Selected from hydrogen, halogen, C 1-6 Alkyl, C 1-6 Haloalkyl and C 3-6 Cycloalkyl;

[0030] R 10 Selected from hydrogen, halogen, cyano, C 1-6 Alkyl and C 1-6 alkyl halide;

[0031] R 11 Selected from hydrogen, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 3-6 Cycloalkyl, 3-6 membered heterocyclic group, -C 1-6 Alkylene-C 3-6 Cycloalkyl and -C 1-6 Alkylene-3-6 membered heterocyclic group;

[0032] R 12 Selected from hydrogen and C 1-6 alkyl;

[0033] m is selected from 0, 1, 2, 3 and 4;

[0034] n is selected from 0, 1, 2, 3 and 4.

[0035] On the other hand, the present invention provides a pharmaceutical composition comprising a prophylactically and / or therapeutically effective amount of a compound of the present invention or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite or prodrug thereof, and one or more pharmaceutically acceptable carriers.

[0036] In another aspect, the present invention provides a kit comprising a compound of the present invention or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite or prodrug thereof, or comprising a pharmaceutical composition of the present invention.

[0037] On the other hand, the present invention provides 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, which targets KRAS G12D protein and is used to degrade KRAS G12D protein.

[0038] On the other hand, the present invention provides 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, for use in preventing and / or treating KRAS G12D-mediated related diseases.

[0039] On the other hand, the present invention provides use 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 in the preparation of a medicament for preventing and / or treating KRAS G12D-mediated related diseases.

[0040] On the other hand, the present invention provides a method for preventing and / or treating KRAS G12D-mediated related diseases, which comprises administering to an individual in need thereof a preventive and / 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.

[0041] In another aspect, the present invention provides methods of preparing the compounds of the present invention. Detailed Description of the Invention

[0043] definition

[0044] 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.

[0045] 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.

[0046] 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), which is optionally substituted by one or more (such as 1, 2 or 3) suitable substituents such as halogen.

[0047] As used herein, the term "alkylene" refers to a divalent saturated aliphatic hydrocarbon group derived from the corresponding "alkyl" by removing one hydrogen atom. For example, as used herein, the term "C 1-6 "Alkylene" refers to an alkylene group having 1 to 6 carbon atoms, for example methylene (-CH2-), ethylene (-CH2CH2-), isopropylene (-CH(CH3)CH2-), which is optionally substituted by one or more (such as 1, 2 or 3) suitable substituents such as halogen.

[0048] As used herein, the term "alkenyl" refers to a straight or branched aliphatic hydrocarbon group having one or more carbon-carbon double bonds. 2-6 The term "alkenyl" refers to an alkenyl group having 2 to 6 carbon atoms and one, two or three (preferably one) carbon-carbon double bonds (such as vinyl, 1-propenyl, 2-propenyl, 2-butenyl, 3-butenyl, 2-pentenyl, 3-pentenyl, 4-pentenyl, 2-hexenyl, 3-hexenyl, 4-hexenyl, 5-hexenyl, 2-methyl-2-propenyl or 4-methyl-3-pentenyl, etc.), which is optionally substituted by one or more (such as 1, 2 or 3) suitable substituents such as halogen.

[0049] As used herein, the term "alkynyl" refers to a straight or branched aliphatic hydrocarbon group having one or more carbon-carbon triple bonds. 2-6 The term "alkynyl" refers to an alkynyl group having 2 to 6 carbon atoms and one, two or three (preferably one) carbon-carbon triple bonds (such as ethynyl, 1-propynyl, 2-propynyl, 2-butynyl, 3-butynyl, 2-pentynyl, 3-pentynyl, 4-pentynyl, 2-hexynyl, 3-hexynyl, 4-hexynyl or 5-hexynyl, etc.), which is optionally substituted by one or more (such as 1, 2 or 3) suitable substituents such as halogen.

[0050] 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., a monocyclic ring such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl or cyclononyl, or a bicyclic ring, including spirocyclic, fused or 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. For example, the term "C 3-6 "Cycloalkyl" refers to a saturated or partially unsaturated non-aromatic monocyclic or polycyclic (such as bicyclic) hydrocarbon ring having 3 to 6 ring carbon atoms (for example cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl), which is optionally substituted by one or more (such as 1, 2 or 3) suitable substituents, for example methyl substituted cyclopropyl.

[0051] As used herein, the term "cycloalkylene" refers to a divalent group derived from a corresponding "cycloalkyl" by removing a hydrogen atom.

[0052] As used herein, the term "halo" or "halogen" group is defined to include fluorine, chlorine, bromine, or iodine.

[0053] As used herein, the term "haloalkyl" refers to an alkyl group substituted by one or more (such as 1, 2 or 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.

[0054] As used herein, the term "heterocyclyl" refers to a saturated or partially unsaturated monocyclic or polycyclic group (including bridged, fused or spirocyclic rings), 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. t (wherein t is 0, 1 or 2) a heteroatom, such as a 3-12 membered heterocyclyl, a 3-7 membered heterocyclyl, a 3-6 membered heterocyclyl, a 5-6 membered heterocyclyl, etc. Representative examples of heterocyclyl include, but are not limited to, oxiranyl, aziridinyl, azetidinyl, oxetanyl, tetrahydrofuranyl, tetrahydropyrrolidinyl, hexahydro-1H-pyrroline, pyrrolidonyl, imidazolidinyl, pyrazolidinyl, tetrahydropyranyl, piperidinyl, morpholinyl, dithianyl, thiomorpholinyl or piperazinyl.

[0055] As used herein, the term "nitrogen-containing heterocyclic group" refers to a group containing at least one nitrogen heteroatom, optionally containing one or more (e.g., 1, 2, 3 or 4) nitrogen-containing heterocyclic groups selected from O or S(O) t (wherein t is 0, 1 or 2), which may include a nitrogen-containing monocyclic heterocyclic group or a polycyclic heterocyclic group (e.g., a nitrogen-containing bridged heterocyclic group, a nitrogen-containing bis-heterocyclic group, a nitrogen-containing spiro heterocyclic group).

[0056] As used herein, the term "oxygen-containing heterocyclic group" refers to a group containing at least one oxygen heteroatom, optionally containing one or more (e.g., 1, 2, 3 or 4) heterocyclic groups selected from N or S(O) t (wherein t is 0, 1 or 2), which may include an oxygen-containing monocyclic heterocyclic group or a polycyclic heterocyclic group (e.g., an oxygen-containing bridged heterocyclic group, an oxygen-containing bis-heterocyclic group, an oxygen-containing spiro heterocyclic group).

[0057] As used herein, the term "heterocyclylene" refers to a divalent group obtained by removing a hydrogen atom from a corresponding "heterocyclyl".

[0058] 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). The aryl group is optionally substituted by one or more (such as 1, 2 or 3) suitable substituents (for example, halogen, -OH, -CN, -NO2 or C 1-6 alkyl, etc.) substituted.

[0059] 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, 2, 3, 4, 5, 6, 9 or 10 carbon atoms, and in each case may be benzo-fused. For example, the heteroaryl or heteroaromatic ring can be selected from 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 pyridyl (ring), pyridazinyl (ring), pyrimidinyl (ring), pyrazinyl (ring), triazinyl (ring) and the like, and benzo derivatives thereof.

[0060] As used herein, the term "heteroarylene" refers to a divalent group derived from a corresponding "heteroaryl" by removing a hydrogen atom.

[0061] 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 present 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.

[0062] 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.

[0063] 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.

[0064] 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.

[0065] Unless otherwise indicated, as used herein, the point of attachment of a substituent may be from any suitable position of the substituent.

[0066] 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.

[0067] 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 15N); oxygen isotopes (e.g. 15 O. 17 O and 18 O); isotopes of phosphorus (such as 32 P); and sulfur isotopes (e.g. 35 S). 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.

[0068] 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 can be produced. Specific individual molecules can also exist as geometric isomers (cis / trans). Similarly, the compounds of the present invention can exist as mixtures of two or more structurally different forms in rapid equilibrium (commonly referred to as tautomers). Representative examples of tautomers include keto-enol tautomers, phenol-ketone tautomers, nitroso-oxime tautomers, imine-enamine tautomers, etc. It is understood that the scope of this 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%).

[0069] 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.

[0070] 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.

[0071] 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.

[0072] 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.

[0073] 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.

[0074] 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.

[0075] 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 themselves may have less pharmacological activity or no pharmacological activity, and when administered to the body or thereon, can be converted into 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 easily converted into the desired therapeutically active compounds in vivo. Additional 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)).

[0076] 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.

[0077] The term "about" means within ±10%, preferably within ±5%, and more preferably within ±2% of the stated numerical value.

[0078] Compound

[0079] The present invention provides a compound of formula (I) or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotope-labeled compound, metabolite or prodrug thereof:

[0080]

[0081] in:

[0082] Ring A is a 3-7 membered heterocyclic group, and Ring A is optionally substituted by one or more R 7 replace;

[0083] Ring E is a 6-12 membered spiro heterocycle, a 6-12 membered heterocycle or a 6-12 membered bridged heterocycle, and is optionally substituted by one or more R 8 replace;

[0084] Ring X is a benzene ring or a 5-6 membered heteroaromatic ring, and ring X is optionally substituted by one or more R 9 replace;

[0085] Ring Y is a 5-6 membered heteroaromatic ring;

[0086] Ring Z is a benzene ring or a 5-6 membered heteroaromatic ring, and is optionally substituted by one or more R 10 replace;

[0087] L 1 is a covalent bond, or is selected from O, S and NR 11 ;

[0088] L 2 is a covalent bond, or is selected from O, S and NR 12 ;

[0089] L 3 Selected from C 1-6 Alkylene, -OC 1-6 Alkylene-, -C 1-6 Alkylene-O- and -C 1-6 Alkylene-OC 1-6 Alkylene-;

[0090] R 1 Selected from hydrogen, halogen, cyano, C 1-6 Alkyl, C 2-6 Alkenyl, C 3-6 Cycloalkyl, C 1-6 Haloalkyl, -OC 1-6 Alkyl, -OC 1-6 Haloalkyl and -OC 3-6 Cycloalkyl;

[0091] R 2 Selected from hydrogen, halogen, cyano, C 1-6 Alkyl, C2-6 Alkenyl, C 3-6 Cycloalkyl, C 1-6 Haloalkyl, -OC 1-6 Alkyl and -OC 1-6 alkyl halide;

[0092] Each R 3 independently selected from hydrogen, halogen, cyano, C 1-6 Alkyl, C 1-6 Haloalkyl and C 3-6 Cycloalkyl;

[0093] R 4 Selected from C 1-6 Alkyl, C 1-6 Haloalkyl and C 3-6 Cycloalkyl;

[0094] R 5a and R 5b are each independently selected from hydrogen, C 1-6 Alkyl, C 1-6 Haloalkyl, -C 1-6 Alkylene -OH and C 3-6 Cycloalkyl; or

[0095] R 5a and R 5b Together with the carbon atoms connected to form C 3-6 Cycloalkyl or 3-6 membered heterocyclic group;

[0096] Each R 6 independently selected from hydrogen, halogen, cyano, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 3-6 Cycloalkyl, -OC 1-6 Alkyl and -OC 1-6 alkyl halide;

[0097] R 7 Selected from hydrogen, C 1-6 Alkyl, C 1-6 Haloalkyl and C 3-6 Cycloalkyl;

[0098] R 8 Selected from hydrogen, halogen, C 1-6 Alkyl, C 1-6 Haloalkyl and C 3-6 Cycloalkyl;

[0099] R 9 Selected from hydrogen, halogen, C 1-6 Alkyl, C 1-6 Haloalkyl and C 3-6 Cycloalkyl;

[0100] R 10 Selected from hydrogen, halogen, cyano, C 1-6 Alkyl and C 1-6 alkyl halide;

[0101] R 11 Selected from hydrogen, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 3-6 Cycloalkyl, 3-6 membered heterocyclic group, -C 1-6 Alkylene-C 3-6 Cycloalkyl and -C 1-6 Alkylene-3-6 membered heterocyclic group;

[0102] R 12 Selected from hydrogen and C 1-6 alkyl;

[0103] m is selected from 0, 1, 2, 3 and 4;

[0104] n is selected from 0, 1, 2, 3 and 4.

[0105] In some embodiments of the compound of Formula (I), L 2 is a covalent bond, or is selected from O, S and NH; Ring E is a 6-12 membered spiro heterocyclic group, a 6-12 membered heterocyclic group or a 6-12 membered bridged heterocyclic group, which is optionally substituted by one or more R 8 Replacement; R 8 is selected from the group consisting of hydrogen, fluorine, chlorine, bromine, methyl, ethyl, isopropyl, trifluoromethyl and cyclopropyl.

[0106] In some embodiments of the compound of Formula (I), L 2 is a covalent bond, or is selected from O, S and NH; Ring E is selected from 6-12 membered nitrogen-containing spiro heterocyclic group, 6-12 membered oxygen-containing spiro heterocyclic group, which is optionally substituted by one or more R 8 Replacement; R 8 is selected from the group consisting of hydrogen, fluorine, chlorine, bromine, methyl, ethyl, isopropyl, trifluoromethyl and cyclopropyl.

[0107] In some embodiments of the compound of Formula (I), L 2 is a covalent bond, or is selected from O, S and NH; Ring E is selected from 6-10 membered nitrogen-containing spiro heterocyclic groups and 6-10 membered oxygen-containing spiro heterocyclic groups, which are optionally substituted by one or more R 8 Replacement; R 8 is selected from the group consisting of hydrogen, fluorine, chlorine, bromine, methyl, ethyl, isopropyl, trifluoromethyl and cyclopropyl.

[0108] In some embodiments of the compound of Formula (I), L 2 is a covalent bond or O; Ring E is selected from

[0109] In some embodiments of the compound of Formula (I), the fragment Selected from

[0110] In some embodiments of the compound of Formula (I), L 1 is a covalent bond, or is selected from O and NR 11 ; Ring A is a 5-membered heterocyclic group, which is optionally substituted by one or more R 7 Replacement; R 7 is selected from hydrogen, methyl, ethyl, isopropyl, trifluoromethyl and cyclopropyl; R 11 Selected from hydrogen, C 1-3 Alkyl, C 1-3 Halogenated alkyl, C 3-6 Cycloalkyl, -C 1-3 Alkylene-C 3-6 Cycloalkyl and -C 1-3 Alkylene-3-6 membered heterocyclic group.

[0111] In some embodiments of the compound of Formula (I), L 1 Selected from O and NR 11 ; Ring A is a 5-membered nitrogen-containing heterocyclic group; R 11 Selected from hydrogen, C 1-3 Alkyl, C 1-3 Haloalkyl and C 3-6 Cycloalkyl.

[0112] In some embodiments of the compound of Formula (I), L 1 Selected from O and NR 11 ; Ring A is pyrrolyl; R 11 is selected from hydrogen, methyl, ethyl, isopropyl, trifluoromethyl and cyclopropyl.

[0113] In some embodiments of the compound of Formula (I), the fragment Selected from

[0114] In some embodiments of the compound of Formula (I), n is 1; R 6 Selected from halogen.

[0115] In some embodiments of the compound of Formula (I), n is 1; R 6 is selected from the group consisting of hydrogen, fluorine, chlorine and bromine.

[0116] In some embodiments of the compound of Formula (I), the fragment for

[0117] In some embodiments of the compound of Formula (I), R 1 is selected from hydrogen, methyl, fluorine, chlorine and methoxy.

[0118] In some embodiments of the compound of Formula (I), R 1 For hydrogen.

[0119] In some embodiments of the compound of Formula (I), R 2 is selected from the group consisting of hydrogen, fluorine, chlorine, bromine, methyl, ethyl, vinyl, trifluoromethyl and cyclopropyl.

[0120] In some embodiments of the compound of Formula (I), R 2 Selected from C 3-6 Cycloalkyl.

[0121] In some embodiments of the compound of Formula (I), R 2 is selected from cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl.

[0122] In some embodiments of the compound of Formula (I), R 2 It is cyclopropyl.

[0123] In some embodiments of the compound of Formula (I), m is selected from 0, 1, and 2; R 3 Each is independently selected from hydrogen, fluorine, chlorine, bromine, methyl, ethyl, isopropyl, trifluoromethyl and cyclopropyl.

[0124] In some embodiments of the compound of Formula (I), m is 2; R 3 are each independently selected from fluoro and methyl.

[0125] In some embodiments of the compound of Formula (I), the fragment for

[0126] In some embodiments of the compound of Formula (I), L 3 Selected from C 1-3 Alkylene, -OC 1-3 Alkylene-, -C 1-3 Alkylene-O- and -C 1-3 Alkylene-OC 1-3 Alkylene-.

[0127] In some embodiments of the compound of Formula (I), L 3 Selected from -CH2-CH2-, -O-CH2-, -CH2-O- and -CH2-O-CH2-.

[0128] In some embodiments of the compound of Formula (I), L 3 It is -O-CH2-.

[0129] In some embodiments of the compound of Formula (I), ring X is phenyl or pyridinyl.

[0130] In some embodiments of the compound of Formula (I), ring X is phenyl.

[0131] In some embodiments of the compound of Formula (I), ring X is

[0132] In some embodiments of the compound of Formula (I), Ring Y is a 5-membered heteroaryl ring.

[0133] In some embodiments of the compound of Formula (I), Ring Y is a 5-membered nitrogen-containing heteroaromatic ring.

[0134] In some embodiments of the compound of Formula (I), Ring Y is pyrazolyl, triazolyl, or isoxazolyl.

[0135] In some embodiments of the compound of Formula (I), Ring Y is triazolyl.

[0136] In some embodiments of the compound of Formula (I), Ring Y is

[0137] In some embodiments of the compound of Formula (I), R 4 Selected from C 1-6 Alkyl and C 3-6 Cycloalkyl.

[0138] In some embodiments of the compound of Formula (I), R 4 Selected from C 3-6 Alkyl and C 3-6 Cycloalkyl.

[0139] In some embodiments of the compound of Formula (I), R 4 Selected from isopropyl, tert-butyl and cyclohexyl.

[0140] In some embodiments of the compound of Formula (I), R 4 Selected from isopropyl and tert-butyl.

[0141] In some embodiments of the compound of Formula (I), R 5a and R 5b Each is independently selected from hydrogen, methyl, ethyl, isopropyl, -CH2-OH and cyclopropyl.

[0142] In some embodiments of the compound of Formula (I), R 5a and R 5b are each independently selected from hydrogen and -C 1-6 Alkylene-OH.

[0143] In some embodiments of the compound of Formula (I), R 5a and R 5b are each independently selected from hydrogen and -CH2-OH.

[0144] In some embodiments of the compound of Formula (I), R 5a is hydrogen, R 5b It is -CH2-OH.

[0145] In some embodiments of the compound of Formula (I), Ring Z is phenyl or pyridinyl, which is optionally replaced by one or more R 10 Replacement; R 10 is selected from the group consisting of hydrogen, fluorine, chlorine, bromine, methyl, ethyl and trifluoromethyl.

[0146] In some embodiments of the compound of Formula (I), Ring Z is phenyl.

[0147] In some embodiments of the compound of Formula (I), Ring Z is

[0148] In some embodiments of the present invention, the compound of formula (I) has the structure of formula (II-a) or formula (II-b):

[0149]

[0150]

[0151] Among them, ring E, L 2 、R 4 and R 11 As defined above.

[0152] In some embodiments of the present invention, the compound of formula (I) has the structure of formula (III-a), formula (III-b), formula (III-c) or formula (III-d):

[0153]

[0154] Among them, ring E, L 2 、R 4 and R 11 As defined above.

[0155] The present invention encompasses compounds obtained by any combination of the various embodiments. Embodiments obtained by combining the technical features or preferred technical features in one embodiment with the technical features or preferred technical features in another embodiment are also within the scope of the present invention.

[0156] In some embodiments of the compound of Formula (I), the compound of Formula (I) is selected from:

[0157]

[0158]

[0159]

[0160]

[0161]

[0162] In some embodiments of the compound of Formula (I), the compound of Formula (I) is selected from:

[0163]

[0164]

[0165]

[0166]

[0167] Preparation method

[0168] When L 3 is -O-CH2-, and ring Y is When, the compound of formula (I) of the present invention can be prepared by the method shown in the following route 1:

[0169] Route 1

[0170]

[0171] in

[0172] Ring A, Ring E, Ring X, Ring Z, R 1 、R 2 、R 3 、R 4 、R 5a 、R 5b 、R 6 、L 1 、L 2 , m and n are as defined above;

[0173] LG 1 LG 2 LG 3 LG 4 LG 5 LG 6 LG 7 and LG 8each independently represents a leaving group, such as a halogen, a methylsulfonate, a thiomethyl, a methylsulfoxide, a methylsulfone, a boronic acid, a borate ester, a methoxy group, or an ethoxy group;

[0174] PG 1 and PG 2 represents a protecting group, for example, benzyloxycarbonyl (Cbz), tert-butyloxycarbonyl (Boc), tetrahydropyranyl (THP), tert-butyl, (S)-1-phenylethanol or benzyl;

[0175] The method comprises the following steps:

[0176] 1) reacting compound I-1 with compound I-2 to obtain compound I-3;

[0177] 2) reacting compound I-3 with compound I-4 to obtain compound I-5;

[0178] 3) reacting compound I-5 with compound I-6 to obtain compound I-7;

[0179] 4) subjecting compound I-7 to a coupling reaction with compound I-8 to obtain compound I-9;

[0180] 5) subjecting compound I-9 to a coupling reaction with compound I-10 to obtain compound I-11;

[0181] 6) subjecting compound I-11 to a deprotection reaction to obtain compound I-12;

[0182] 7) reacting compound I-12 with compound I-13 to obtain compound I-14;

[0183] 8) reacting compound 1-14 with compound 1-15 to obtain compound 1-16; and

[0184] 9) Compound I-16 is subjected to a deprotection reaction to obtain Compound I.

[0185] The reaction in step (1) is preferably carried out in a suitable organic solvent and in the presence of a suitable organic base. The organic solvent may be selected from N,N-dimethylformamide, dimethyl sulfoxide, N,N-dimethylacetamide, tetrahydrofuran, 1,4-dioxane, and any combination thereof, preferably a combination of tetrahydrofuran and 1,4-dioxane. The organic base may be selected from N,N-diisopropylethylamine and triethylamine, preferably N,N-diisopropylethylamine. The reaction is preferably carried out at a temperature of -50°C to 80°C for 2-24 hours.

[0186] The reaction in step (2) is preferably carried out in a suitable organic solvent and in the presence of a suitable base. The organic solvent may be selected from acetonitrile, N,N-dimethylformamide, dimethyl sulfoxide, N,N-dimethylacetamide, tetrahydrofuran, 1,4-dioxane, and any combination thereof, preferably acetonitrile. The base may be selected from N,N-diisopropylethylamine, triethylamine, potassium carbonate, triethylenediamine, cesium carbonate, and any combination thereof, preferably a combination of triethylenediamine and cesium carbonate. The reaction is preferably carried out at a temperature of 20°C to 100°C for 2 to 48 hours.

[0187] The reaction in step (3) is preferably carried out in a suitable organic solvent and in the presence of a suitable base. The base may be selected from sodium hydride, sodium tert-butoxide, or potassium tert-butoxide, preferably potassium tert-butoxide. The organic solvent may be selected from tetrahydrofuran, dichloromethane, toluene, 1,4-dioxane, N,N-dimethylformamide, and any combination thereof, preferably tetrahydrofuran. The reaction is preferably carried out at a temperature of 0-100° C. for 2-16 hours.

[0188] The coupling reaction of the above step (4) is preferably carried out in a suitable solvent and in the presence of a metal catalyst and a base. The metal catalyst can be a palladium metal catalyst, such as tris (dibenzylideneacetone) dipalladium, [1,1'-bis (diphenylphosphino) ferrocene] dichloropalladium, methanesulfonic acid [n-butyldi (1-adamantyl) phosphine] (2-amino-1,1'-biphenyl-2-yl) palladium (II), tetrakistriphenylphosphine palladium and palladium acetate, preferably [1,1'-bis (diphenylphosphino) ferrocene] dichloropalladium. The base can be an organic base or an inorganic base, for example, selected from sodium tert-butoxide, potassium carbonate, potassium phosphate, cesium carbonate and sodium carbonate, preferably potassium phosphate. The solvent can be selected from N,N-dimethylformamide, N-methylpyrrolidone, toluene, ethanol, ethylene glycol dimethyl ether, water, 1,4-dioxane and any combination thereof, preferably a combination of 1,4-dioxane and water. The reaction is preferably carried out at a temperature of 50-120°C for 2-16 hours.

[0189] The coupling reaction of step (5) is preferably carried out in a suitable solvent in the presence of a metal catalyst, a phosphine ligand and a base. The metal catalyst can be a palladium metal catalyst, such as tris(dibenzylideneacetone)dipalladium, [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium, methanesulfonic acid [n-butyldi(1-adamantyl)phosphine](2-amino-1,1'-biphenyl-2-yl)palladium(II), tetrakistriphenylphosphine palladium and palladium acetate, preferably tris(dibenzylideneacetone)dipalladium. The phosphine ligand can be selected from 2-dicyclohexylphosphine-2,6-dimethoxy-biphenyl, 2-dicyclohexylphosphine-2',4',6'-triisopropylbiphenyl, 2-dicyclohexylphosphine-2',6'-diisopropyl-1,1'-biphenyl, 1,1'-binaphthyl-2,2'-bisdiphenylphosphine and 4,5-bisdiphenylphosphine-9,9-dimethylxanthene, preferably 2-dicyclohexylphosphine-2,6-dimethoxy-biphenyl. The base can be an organic base or an inorganic base, for example, selected from sodium tert-butoxide, potassium carbonate, potassium phosphate, cesium carbonate and sodium carbonate, preferably potassium phosphate. The solvent can be selected from N,N-dimethylformamide, N-methylpyrrolidone, toluene, ethanol, ethylene glycol dimethyl ether, water, 1,4-dioxane and any combination thereof, preferably a combination of 1,4-dioxane and water. The reaction is preferably carried out at a temperature of 50-120°C for 2-16 hours.

[0190] The deprotection reaction in step (6) is preferably carried out in a suitable organic solvent and in the presence of a suitable metal catalyst. The organic solvent may be selected from methanol, ethanol, isopropanol, ethyl acetate, tetrahydrofuran, and any combination thereof, preferably methanol. The metal catalyst is a palladium metal catalyst or a platinum metal catalyst, such as palladium carbon, palladium hydroxide, platinum dioxide, preferably palladium carbon. The reduction reaction is preferably carried out in the presence of hydrogen. The reaction is preferably carried out at a temperature of 0-80°C for 1-24 hours.

[0191] The reaction in step (7) is preferably carried out in a suitable organic solvent and in the presence of a suitable organic base. The organic solvent may be selected from dichloromethane, tetrahydrofuran, acetonitrile, N,N-dimethylformamide, and any combination thereof, preferably N,N-dimethylformamide. The base may be selected from N,N-diisopropylethylamine, triethylamine, potassium carbonate, triethylenediamine, cesium carbonate, and any combination thereof. Preferably, it is cesium carbonate. The reaction is preferably carried out at a temperature of 0-100° C. for 1-12 hours.

[0192] The reaction of step (8) is preferably carried out in a suitable solvent and in the presence of a suitable reagent. The solvent may be selected from dichloromethane, tetrahydrofuran, acetonitrile, dimethyl sulfoxide, N,N-dimethylformamide, water, and any combination thereof, preferably a combination of dimethyl sulfoxide and water. The reagent may be selected from copper sulfate pentahydrate, cuprous iodide, cuprous bromide, cuprous chloride, copper acetate, copper oxide, sodium ascorbate, and any combination thereof, preferably a combination of copper sulfate pentahydrate and sodium ascorbate. The reaction is preferably carried out at a temperature of 0-100° C. for 1-12 hours.

[0193] The deprotection reaction in step (9) is preferably carried out in a suitable organic solvent and in the presence of a suitable deprotection agent. The organic solvent may be selected from ethyl acetate, toluene, acetonitrile, 1,4-dioxane, dichloromethane, and any combination thereof, preferably dichloromethane. The deprotection agent may be selected from hydrochloric acid, hydrobromic acid, p-toluenesulfonic acid, methanesulfonic acid, trifluoroacetic acid, aluminum chloride, and trimethylsilyl iodide, preferably trifluoroacetic acid. The reaction is preferably carried out at a temperature of 0-80° C. for 1-12 hours.

[0194] When L 3 is -O-CH2-, and ring Y is The present invention provides a second method for preparing a compound of formula (I), comprising the following steps:

[0195] Route 2

[0196]

[0197]

[0198] in

[0199] Ring A, Ring E, Ring X, Ring Z, R 1 、R 2 、R 3 、R 4 、R 5a 、R 5b 、R 6 、L 1 、L 2 , m and n are as defined above;

[0200] LG 1 LG 2 LG 3 LG 4 LG 5 LG 6 LG 7 and LG 8each independently represents a leaving group, such as a halogen, a methylsulfonate, a thiomethyl, a methylsulfoxide, a methylsulfone, a boronic acid, a borate ester, a methoxy group, or an ethoxy group;

[0201] PG 1 and PG 2 represents a protecting group, for example, benzyloxycarbonyl (Cbz), tert-butyloxycarbonyl (Boc), tetrahydropyranyl (THP), tert-butyl, (S)-1-phenylethanol or benzyl;

[0202] The method comprises the following steps:

[0203] 1) reacting compound I-1 to obtain compound I-2a;

[0204] 2) reacting compound I-2a to obtain compound I-3a;

[0205] 3) reacting compound I-3a to obtain compound I-4a;

[0206] 4) reacting compound I-4a with compound I-6 to obtain compound I-5a;

[0207] 5) coupling reaction of compound I-5a with compound I-8 to obtain compound I-6a;

[0208] 6) coupling reaction of compound I-6a with compound I-10 to obtain compound I-7a;

[0209] 7) Compound I-7a is reacted to obtain compound I-8a

[0210] 8) subjecting compound I-8a to a deprotection reaction to obtain compound I-9a;

[0211] 9) reacting compound I-9a with compound I-13 to obtain compound I-10a;

[0212] 10) reacting compound I-10a to obtain compound I-11a;

[0213] 11) reacting compound I-11a to obtain compound I-12a;

[0214] 12) reacting compound I-12a to obtain compound I-14;

[0215] 13) reacting compound 1-14 with compound 1-15 to obtain compound 1-16; and

[0216] 14) Compound I-16 is subjected to a deprotection reaction to obtain Compound I.

[0217] The reaction in step (1) is preferably carried out in a suitable solvent and in the presence of a suitable base. The solvent may be selected from N,N-dimethylformamide, dimethyl sulfoxide, N,N-dimethylacetamide, tetrahydrofuran, 1,4-dioxane, water, and any combination thereof, preferably a combination of tetrahydrofuran and water. The base may be selected from lithium hydroxide, sodium hydroxide, and potassium hydroxide, preferably sodium hydroxide. The reaction is preferably carried out at a temperature of -50°C to 80°C for 2-24 hours.

[0218] The reaction in step (2) is preferably carried out in a suitable organic solvent and in the presence of a suitable reagent. The organic solvent may be selected from N,N-dimethylformamide, dimethyl sulfoxide, N,N-dimethylacetamide, tetrahydrofuran, 1,4-dioxane, and any combination thereof, preferably tetrahydrofuran. The reagent may be selected from 2-tert-butyl-1,3-diisopropylisourea and di-tert-butyl dicarbonate, preferably 2-tert-butyl-1,3-diisopropylisourea. The reaction is preferably carried out at a temperature of 0°C to 100°C for 2-24 hours.

[0219] The reaction of step (3) is preferably carried out in a suitable organic solvent and in the presence of a suitable base and a suitable reagent. The organic solvent may be selected from acetonitrile, N,N-dimethylformamide, dimethyl sulfoxide, dichloromethane, tetrahydrofuran, 1,4-dioxane, and any combination thereof, preferably dichloromethane. The base may be selected from N,N-diisopropylethylamine, triethylamine, potassium carbonate, triethylenediamine, cesium carbonate, and any combination thereof, preferably triethylenediamine. The reagent may be selected from ethanethiol and sodium ethanethiolate, preferably ethanethiol. The reaction is preferably carried out at a temperature of 0°C-100°C for 2-48 hours.

[0220] The reaction in step (4) is preferably carried out in a suitable organic solvent and in the presence of a suitable base. The base may be selected from sodium hydride, sodium tert-butoxide, and potassium tert-butoxide, preferably potassium tert-butoxide. The organic solvent may be selected from tetrahydrofuran, dichloromethane, toluene, 1,4-dioxane, N,N-dimethylformamide, and any combination thereof, preferably tetrahydrofuran. The reaction is preferably carried out at a temperature of 0-100° C. for 2-16 hours.

[0221] The coupling reaction of the above-mentioned step (5) is preferably carried out in a suitable solvent and in the presence of a metal catalyst and a base. The metal catalyst can be a palladium metal catalyst, such as tris (dibenzylideneacetone) dipalladium, [1,1'-bis (diphenylphosphino) ferrocene] palladium dichloride, methanesulfonic acid [n-butyldi (1-adamantyl) phosphine] (2-amino-1,1'-biphenyl-2-yl) palladium (II), tetrakistriphenylphosphine palladium and palladium acetate, preferably [1,1'-bis (diphenylphosphino) ferrocene] palladium dichloride. The base can be an organic base or an inorganic base, for example, selected from sodium tert-butoxide, potassium carbonate, potassium phosphate, cesium carbonate and sodium carbonate, preferably potassium phosphate. The solvent can be selected from N,N-dimethylformamide, N-methylpyrrolidone, toluene, acetonitrile, ethanol, ethylene glycol dimethyl ether, water, 1,4-dioxane and any combination thereof, preferably a combination of acetonitrile and water. The reaction is preferably carried out at a temperature of 30-120°C for 2-16 hours.

[0222] The coupling reaction of step (6) is preferably carried out in a suitable solvent in the presence of a metal catalyst, a phosphine ligand and a base. The metal catalyst can be a palladium metal catalyst, such as tris(dibenzylideneacetone)dipalladium, [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium, methanesulfonic acid [n-butyldi(1-adamantyl)phosphine](2-amino-1,1'-biphenyl-2-yl)palladium(II), tetrakistriphenylphosphine palladium and palladium acetate, preferably tris(dibenzylideneacetone)dipalladium. The phosphine ligand can be selected from 2-dicyclohexylphosphine-2,6-dimethoxy-biphenyl, 2-dicyclohexylphosphine-2',4',6'-triisopropylbiphenyl, 2-dicyclohexylphosphine-2',6'-diisopropyl-1,1'-biphenyl, 1,1'-binaphthyl-2,2'-bisdiphenylphosphine and 4,5-bisdiphenylphosphine-9,9-dimethylxanthene, preferably 2-dicyclohexylphosphine-2,6-dimethoxy-biphenyl. The base can be an organic base or an inorganic base, for example, selected from sodium tert-butoxide, potassium carbonate, potassium phosphate, cesium carbonate and sodium carbonate, preferably potassium phosphate. The solvent can be selected from N,N-dimethylformamide, N-methylpyrrolidone, toluene, ethanol, ethylene glycol dimethyl ether, water, 1,4-dioxane and any combination thereof, preferably a combination of 1,4-dioxane and water. The reaction is preferably carried out at a temperature of 50-120°C for 2-16 hours.

[0223] The reaction in step (7) is preferably carried out in a suitable organic solvent and in the presence of a suitable oxidizing agent. The organic solvent may be selected from ethyl acetate, methanol, 1,4-dioxane, dichloromethane, and any combination thereof, preferably dichloromethane. The oxidizing agent may be selected from hydrogen peroxide, m-chloroperbenzoic acid, and potassium persulfate. M-chloroperbenzoic acid is preferred. The reaction is preferably carried out at a temperature of 0-60° C. for 1-16 hours.

[0224] The deprotection reaction in step (8) is preferably carried out in a suitable organic solvent and in the presence of a suitable metal catalyst. The organic solvent may be selected from methanol, ethanol, isopropanol, ethyl acetate, tetrahydrofuran, and any combination thereof, preferably methanol. The metal catalyst is a palladium metal catalyst or a platinum metal catalyst, such as palladium carbon, palladium hydroxide, platinum dioxide, preferably palladium carbon. The reduction reaction is preferably carried out in the presence of hydrogen. The reaction is preferably carried out at a temperature of 0-80°C for 1-24 hours.

[0225] The reaction in step (9) is preferably carried out in a suitable organic solvent and in the presence of a suitable organic base. The organic solvent may be selected from dichloromethane, tetrahydrofuran, acetonitrile, N,N-dimethylformamide, and any combination thereof, preferably N,N-dimethylformamide. The base may be selected from N,N-diisopropylethylamine, triethylamine, potassium carbonate, triethylenediamine, cesium carbonate, and any combination thereof. Preferably, it is cesium carbonate. The reaction is preferably carried out at a temperature of 0-100° C. for 1-12 hours.

[0226] The reaction in step (10) is preferably carried out in a suitable organic solvent and in the presence of a suitable base. The organic solvent may be selected from acetonitrile, toluene, N,N-dimethylformamide, dimethyl sulfoxide, N,N-dimethylacetamide, tetrahydrofuran, 1,4-dioxane, and any combination thereof, preferably tetrahydrofuran. The base may be selected from sodium tert-butoxide, potassium tert-butoxide, and sodium hydride, preferably potassium tert-butoxide. The reaction is preferably carried out at a temperature of 0-60° C. for 2-24 hours.

[0227] The reaction in step (11) is preferably carried out in a suitable organic solvent and in the presence of a suitable acid. The organic solvent may be selected from tetrahydrofuran, acetonitrile, 1,4-dioxane, dichloromethane, and any combination thereof, preferably tetrahydrofuran. The acid may be selected from hydrochloric acid, hydrobromic acid, p-toluenesulfonic acid, methanesulfonic acid, and trifluoroacetic acid, preferably p-toluenesulfonic acid. The reaction is preferably carried out at a temperature of 0-80° C. for 0.5-12 hours.

[0228] The reaction of step (12) is preferably carried out in a suitable organic solvent and in the presence of a suitable base and a suitable reagent. The organic solvent may be selected from tetrahydrofuran, acetonitrile, 1,4-dioxane, N,N-dimethylformamide, dimethyl sulfoxide, dichloromethane and any combination thereof, preferably tetrahydrofuran or N,N-dimethylformamide. The base may be selected from cesium carbonate, potassium carbonate, triethylamine and N,N-diisopropylethylamine, preferably cesium carbonate. The reagent may be selected from 1H-benzotriazole-1-yloxytripyrrolidino hexafluorophosphate, 1-hydroxybenzotriazole and Carter condensation agent, preferably 1H-benzotriazole-1-yloxytripyrrolidino hexafluorophosphate. The reaction is preferably carried out at a temperature of 0-80°C for 1-24 hours.

[0229] The reaction of step (13) is preferably carried out in a suitable solvent and in the presence of a suitable reagent. The solvent may be selected from dichloromethane, tetrahydrofuran, acetonitrile, dimethyl sulfoxide, N,N-dimethylformamide, water, and any combination thereof, preferably a combination of dimethyl sulfoxide and water. The reagent may be selected from copper sulfate pentahydrate, cuprous iodide, cuprous bromide, cuprous chloride, copper acetate, copper oxide, sodium ascorbate, and any combination thereof, preferably a combination of copper sulfate pentahydrate and sodium ascorbate. The reaction is preferably carried out at a temperature of 0-100° C. for 1-12 hours.

[0230] The deprotection reaction in step (14) is preferably carried out in a suitable organic solvent and in the presence of a suitable deprotection agent. The organic solvent may be selected from ethyl acetate, toluene, acetonitrile, 1,4-dioxane, dichloromethane, and any combination thereof, preferably dichloromethane. The deprotection agent may be selected from hydrochloric acid, hydrobromic acid, p-toluenesulfonic acid, methanesulfonic acid, trifluoroacetic acid, aluminum chloride, and trimethylsilyl iodide, preferably trifluoroacetic acid. The reaction is preferably carried out at a temperature of 0-80° C. for 1-12 hours.

[0231] Pharmaceutical compositions and kits

[0232] Another object of the present invention is to provide a pharmaceutical composition comprising a preventively and / 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, and one or more pharmaceutically acceptable carriers.

[0233] Another object of the present invention is to provide a drug kit comprising the compound of the present invention or its pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotope-labeled compound, metabolite or prodrug, or the pharmaceutical composition of the present invention.

[0234] 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.

[0235] 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.

[0236] The pharmaceutical composition may 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 may also be in the form of liposomes, microspheres, or other dosage forms.

[0237] 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.

[0238] 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 0.01-800 mg, preferably 0.05-500 mg, more preferably 0.1-350 mg, particularly preferably 0.5-100 mg.

[0239] 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, N-oxide, isotopically labeled compound, metabolite or prodrug thereof with one or more pharmaceutically acceptable carriers.

[0240] Treatment methods and uses

[0241] Another object of the present invention is to provide 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, for use in preventing and / or treating KRAS G12D-mediated related diseases.

[0242] Another object of the present invention is to provide use of the compound of the present invention or its pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotope-labeled compound, metabolite or prodrug, or the pharmaceutical composition of the present invention in the preparation of a medicament for preventing and / or treating KRAS G12D-mediated related diseases.

[0243] Another object of the present invention is to provide a method for preventing and / or treating KRAS G12D-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.

[0244] According to some embodiments of the present invention, the KRAS G12D-mediated related disease is a tumor or cancer.

[0245] 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.

[0246] 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.

[0247] 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.

[0248] The term "prevention" as used herein includes inhibition and delay of the onset of a disease, and includes not only prevention before the development of a disease but also prevention of recurrence of a disease after treatment.

[0249] The terms "treat," "treat," "treat," or "treating" as used herein means to reverse, alleviate, or eliminate the progression of the disorder or condition to which such terms apply, or one or more symptoms of such disorder or condition.

[0250] 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

[0251] The embodiments of the present invention are described in detail below with reference to the examples. However, those skilled in the art will appreciate that the following examples are only used to illustrate the present invention and should not be considered as limiting the scope of the present invention.

[0252] In the examples, if the specific conditions are not specified, all experiments were carried out under conventional conditions or those recommended by the manufacturer. All reagents or instruments used, if the manufacturer is not specified, are commercially available conventional products.

[0253] The structure of the compound was determined by nuclear magnetic resonance ( 1 H NMR) and / or mass spectrometry (MS).

[0254] 1 H NMR measurements were performed on a JEOL Eclipse 400 nuclear magnetometer. The solvents used were deuterated methanol (CD3OD), deuterated chloroform (CDCl3), or hexadeuterated dimethyl sulfoxide (DMSO-d6). The internal standard was tetramethylsilane (TMS). Chemical shifts (δ) were given in parts per million (ppm).

[0255] The MS measuring instrument is an Agilent (ESI) mass spectrometer, manufactured by Agilent, model: Agilent 6120B.

[0256] The preparative HPLC conditions were as follows:

[0257] Instrument model: Agilent 1260; chromatographic column: Waters SunFire Prep C18 OBD (19 mm × 150 mm × 5.0 μm); column temperature: 25°C; flow rate: 20.0 mL / min; detection wavelength: 214 nm; elution gradient: (0 min: 10% A, 90% B; 16.0 min: 90% A, 10% B); mobile phase A: acetonitrile; mobile phase B: 0.05% formic acid in water.

[0258] The reverse phase column used was an Agela C18 reverse flash column (20-35 μm / 100A).

[0259] Aluminum plates (20×20 cm) produced by Merck were used for thin layer chromatography silica gel plates (TLC), and the specifications used for thin layer chromatography separation and purification were GF 254 (1 mm) produced in Yantai.

[0260] The reaction is monitored by thin layer chromatography (TLC) or LC-MS; the developing solvent systems used include: dichloromethane and methanol system, n-hexane and ethyl acetate system, and petroleum ether and ethyl acetate system. The volume ratio of the solvent is adjusted according to the polarity of the compound or by adding triethylamine.

[0261] Column chromatography generally uses 200-300 mesh silica gel as a carrier. Eluent systems include: dichloromethane and methanol systems, and petroleum ether and ethyl acetate systems. The volume ratio of the solvent is adjusted according to the polarity of the compound, and a small amount of triethylamine can also be added for adjustment.

[0262] Unless otherwise specified, the reaction temperature was room temperature (20°C-35°C).

[0263] The reagents used in the examples were purchased from Acros Organics, Aldrich Chemical Company, Teber Chemical and other companies.

[0264] The meanings of the abbreviations used in this document are shown in the following table.

[0265] abbreviation meaning TLC Thin layer chromatography LC-MS Liquid chromatography-mass spectrometry DMF N,N-Dimethylformamide DMSO dimethyl sulfoxide <![CDATA[Pd(dppf)Cl2]]> [1,1'-Bis(diphenylphosphino)ferrocene]palladium dichloride <![CDATA[CD3OD]]> Deuterated methanol <![CDATA[CDCl3]]> Deuterated chloroform <![CDATA[DMSO-d6]]> Hexadeuterated dimethyl sulfoxide TMS Tetramethylsilane NMR Nuclear magnetic resonance imaging MS Mass spectrometry s singlet d Doublet t Triplet q quartet dd Double doublet m multiplet br broad J Coupling constant Hz hertz

[0266] Preparation of intermediates:

[0267] Intermediate Preparation Example 1: Preparation of (2S,4R)-1-((S)-2-azido-3-isopropyl)-N-((R)-1-(2-fluoro-[1,1-biphenyl]-4-yl)-2-hydroxyethyl)-4-hydroxypyrrolidine-2-carboxamide

[0268]

[0269] Step 1: Preparation of tert-butyl (R)-(1-(2-fluoro-(1,1-biphenyl)-4-yl)-2-hydroxyethyl)carbamate

[0270] Dissolve tert-butyl N-[(R)-1-(4-bromophenyl)-2-hydroxyethyl]carbamate (5 g, 15.3 mmol) and 2-fluorophenylboronic acid (2.8 g, 19.9 mmol) in 1,4-dioxane (40 mL) and water (5 mL). Add Pd(dppf)Cl2 (572.6 mg, 766.9 μmol) and potassium carbonate (4.3 g, 30.7 mmol) and react at 95°C for 3 hours. Dilute the reaction solution with water (30 mL) and extract twice with ethyl acetate (30 mL). The organic phase is washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue is purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 1 / 1, volume ratio) to obtain the title compound (5 g, yield: 93.5%).

[0271] MS m / z(ESI):332.2[M+H] + .

[0272] Step 2: Preparation of (R)-(1-(2-fluoro-(1,1-biphenyl)-4-yl)-2-hydroxyethyl)amine

[0273] Dissolve tert-butyl (R)-(1-(2-fluoro-(1,1-biphenyl)-4-yl)-2-hydroxyethyl)carbamate (5 g, 14.3 mmol) in dichloromethane (30 mL). Add a solution of hydrogen chloride in 1,4-dioxane (30 mL) and react at 25°C for 1 hour. Concentrate the reaction mixture to obtain the title compound (3.9 g, yield: 96.6%).

[0274] MS m / z(ESI):232.2[M+H] + .

[0275] Step 3: Preparation of tert-butyl (2S,4R)-2-(((R)-1-(2-fluoro-[1,1-biphenyl]-4-yl)-2-hydroxyethyl)carbamoyl)-4-hydroxypyrrolidine-1-carboxylate

[0276] (R)-(1-(2-Fluoro-(1,1-biphenyl)-4-yl)-2-hydroxyethyl)amine (3.9 g, 13.8 mmol) and N,N-diisopropylethylamine (4.5 g, 34.6 mmol) were dissolved in DMF (30 mL), and (2S,4R)-1-tert-butylcarbonyl-4-hydroxypyrrolidine-2-carboxylic acid (3.8 g, 13.9 mmol) and (7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (8.3 g, 20.8 mmol) were added. The mixture was reacted at 25°C for 3 hours. The reaction solution was purified by reverse-phase column chromatography (eluent: water (containing 0.05% formic acid) / acetonitrile = 1 / 1, volume ratio) to give the title compound (5.6 g, yield: 86.5%).

[0277] MS m / z(ESI):445.2[M+H] + .

[0278] Step 4: Preparation of (2S,4R)-N-((R)-1-(2-fluoro-[1,1-biphenyl]-4-yl)-2-hydroxyethyl)-4-hydroxypyrrolidine-2-carboxamide

[0279] Dissolve tert-butyl (2S,4R)-2-(((R)-1-(2-fluoro-[1,1-biphenyl]-4-yl)-2-hydroxyethyl)carbamoyl)-4-hydroxypyrrolidine-1-carboxylate (5.6 g, 12.0 mmol) in dichloromethane (30 mL). Add a solution of hydrogen chloride in 1,4-dioxane (30 mL) and react at 25°C for 1 hour. Concentrate the reaction mixture to obtain the title compound (4.6 g, yield: 95.9%).

[0280] MS m / z(ESI):345.2[M+H] + .

[0281] Step 5: Preparation of tert-butyl (S)-1-((2S,4R)-2-(((R)-1-(2'-fluoro-[1,1'-biphenyl]-4-yl)-2-hydroxyethyl)carbamoyl)-4-hydroxypyrrolidin-1-yl)-3-methyl-1-oxobutan-2-yl)carbamate

[0282] (2S,4R)-N-((R)-1-(2-fluoro-[1,1-biphenyl]-4-yl)-2-hydroxyethyl)-4-hydroxypyrrolidine-2-carboxamide (4.6 g, 11.5 mmol) and N,N-diisopropylethylamine (4.5 g, 34.6 mmol) were dissolved in DMF (30 mL), and (2S)-2-(tert-butylcarbonylamino)-3-methylbutanoic acid (2.62 g, 12.1 mmol) and (7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (6.9 g, 17.3 mmol) were added. The mixture was reacted at 25°C for 3 hours. The reaction solution was purified by reverse-phase column chromatography (eluent: water (containing 0.05% formic acid) / acetonitrile = 1 / 1, volume ratio) to give the title compound (5.8 g, yield: 88.3%).

[0283] MS m / z(ESI):544.3[M+H] + .

[0284] Step 6: Preparation of (2S,4R)-1-(L-valine)-N-((R)-1-[2'-fluoro-[1,1'-biphenyl]-4-yl]-2-hydroxyethyl)-4-hydroxypyrrolidine-2-carboxamide

[0285] Tert-butyl (S)-1-((2S,4R)-2-(((R)-1-(2'-fluoro-[1,1'-biphenyl]-4-yl)-2-hydroxyethyl)carbamoyl)-4-hydroxypyrrolidin-1-yl)-3-methyl-1-oxobutan-2-yl)carbamate (5.8 g, 10.1 mmol) was dissolved in dichloromethane (30 mL). A solution of hydrogen chloride in 1,4-dioxane (30 mL) was added and the mixture was reacted at 25°C for 1 hour. The reaction mixture was concentrated to obtain the title compound (4.9 g, yield: 95.7%).

[0286] MS m / z(ESI):444.2[M+H] + .

[0287] Step 7: Preparation of (2S,4R)-1-((S)-2-azido-3-isopropyl)-N-((R)-1-(2-fluoro-[1,1-biphenyl]-4-yl)-2-hydroxyethyl)-4-hydroxypyrrolidine-2-carboxamide

[0288] (2S,4R)-1-(L-valine)-N-((R)-1-[2'-fluoro-[1,1'-biphenyl]-4-yl]-2-hydroxyethyl)-4-hydroxypyrrolidine-2-carboxamide (4.8 g, 9.5 mmol) was dissolved in methanol (20 mL). Potassium carbonate (4.6 g, 33.3 mmol) and copper sulfate pentahydrate (242.0 mg, 950.0 μmol) were added, followed by 1H-imidazole-1-sulfonyl azide (3.1 g, 14.3 mmol). The mixture was stirred at 25°C for 12 hours. The reaction mixture was diluted with water (30 mL) and extracted twice with ethyl acetate (30 mL). The organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: dichloromethane / methanol = 10 / 1, volume ratio) to give the title compound (3.5 g, yield: 74.5%).

[0289] MS m / z(ESI):470.2[M+H] + .

[0290] Intermediate Preparation Example 2: Preparation of 2-((2-oxaspiro[3.3]hept-6-yl)oxy)-6-cyclopropyl-8-((4-ethynylbenzyl)oxy)-7-(6-fluoro-5-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)quinazolin-4-ol

[0291]

[0292] Step 1: Preparation of 7-bromo-2-chloro-8-fluoro-6-iodoquinazolin-4-ol

[0293] Dissolve 7-bromo-2,4-dichloro-8-fluoro-6-iodoquinazoline (13.0 g, 30.8 mmol) in tetrahydrofuran (100 mL). Cool the reaction system to 0°C in an ice-water bath. Add 1M aqueous sodium hydroxide solution (62 mL) dropwise and stir at 0°C for 2 hours. Slowly pour the reaction solution into 1M hydrochloric acid (360 mL) and stir at 25°C for 0.5 hours. Solid precipitates, which is filtered, rinsed with water (200 mL), and then dried to obtain the title compound (12.4 g, yield: 99.9%).

[0294] MS m / z(ESI):403.1[M+H] + .

[0295] Step 2: Preparation of 7-bromo-4-(tert-butoxy)-2-chloro-8-fluoro-6-iodoquinazoline

[0296] Dissolve 7-bromo-2-chloro-8-fluoro-6-iodoquinazolin-4-ol (8.0 g, 19.8 mmol) in tetrahydrofuran (200 mL), heat to 65°C and stir for 15 minutes. Add tert-butyl N,N'-diisopropylcarbamoyl ester (23.8 g, 118.8 mmol) dropwise, and stir at 65°C for 2 hours. Cool the reaction mixture to room temperature, filter, and rinse the solid with tetrahydrofuran (50 mL). The filtrate is concentrated under reduced pressure, and anhydrous methanol (70 mL) is added to the residue with stirring to precipitate a solid. Filter the filter cake, rinse with anhydrous methanol (30 mL), and collect and dry it to obtain the title compound (7.0 g, yield: 76.8%).

[0297] MS m / z(ESI):459.3[M+H] + .

[0298] Step 3: Preparation of 7-bromo-4-(tert-butoxy)-2-(ethylthio)-8-fluoro-6-iodoquinazoline

[0299] 7-Bromo-4-(tert-butoxy)-2-chloro-8-fluoro-6-iodoquinazoline (7.0 g, 15.2 mmol) was dissolved in dichloromethane (100 mL). Triethylenediamine (2.5 g, 22.8 mmol) and ethanethiol (1.4 g, 22.8 mmol) were added sequentially, and the mixture was stirred at 25°C for 12 hours. Water (100 mL) was slowly added to the reaction solution, and the layers were separated. The aqueous phase was extracted twice with dichloromethane (50 mL). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain the title compound (7.3 g, yield: 98.7%).

[0300] MS m / z(ESI):485.1[M+H] + .

[0301] Step 4: Preparation of (S)-7-bromo-4-(tert-butoxy)-2-(ethylthio)-6-iodo-8-(1-phenylethoxy)quinazoline

[0302] 7-Bromo-4-(tert-butoxy)-2-(ethylthio)-8-fluoro-6-iodoquinazoline (7.3 g, 15.0 mmol) was dissolved in tetrahydrofuran (100 mL). Potassium tert-butoxide (2.5 g, 22.6 mmol) and (S)-1-phenylethanol-1-ol (2.4 g, 19.6 mmol) were added sequentially, and the mixture was stirred at 25°C for 2 hours. The reaction solution was diluted with ethyl acetate (500 mL) and washed twice with saturated brine (100 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: n-hexane / ethyl acetate = 25 / 1, volume ratio) to obtain the title compound (8.6 g, yield: 96.9%).

[0303] MS m / z(ESI):587.1[M+H] + .

[0304] Step 5: Preparation of (S)-7-bromo-4-(tert-butoxy)-6-cyclopropyl-2-(ethylthio)-8-(1-phenylethoxy)quinazoline

[0305] (S)-7-Bromo-4-(tert-butoxy)-2-(ethylthio)-6-iodo-8-(1-phenylethoxy)quinazoline (8.6 g, 14.6 mmol) was dissolved in acetonitrile (100 mL) and water (20 mL). Cyclopropylboronic acid (1.8 g, 20.5 mmol), Pd(dppf)Cl2 (1.2 g, 1.5 mmol), and potassium phosphate (12.4 g, 58.6 mmol) were added sequentially, and the mixture was stirred at 90°C for 4 hours. The reaction solution was cooled to room temperature, diluted with ethyl acetate (300 mL), and washed twice with saturated brine (50 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: n-hexane / ethyl acetate = 50 / 1, volume ratio) to obtain the title compound (6.0 g, yield: 81.7%).

[0306] MS m / z(ESI):501.2[M+H] + .

[0307] Step 6: Preparation of 4-(tert-butoxy)-6-cyclopropyl-2-(ethylthio)-7-(6-fluoro-5-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)-8-((S)-1-phenylethoxy)quinazoline

[0308] (S)-7-Bromo-4-(tert-butoxy)-6-cyclopropyl-2-(ethylthio)-8-(1-phenylethoxy)quinazoline (6.0 g, 12.0 mmol) was dissolved in 1,4-dioxane (60 mL) and water (6 mL). 6-Fluoro-5-methyl-1-(tetrahydro-2H-pyran-2-yl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indazole (5.2 g, 14.4 mmol), tris(dibenzylideneacetone)dipalladium (2.2 g, 2.4 mmol), 2-dicyclohexylphosphino-2',6'-dimethoxybiphenyl (1.0 g, 2.4 mmol) and potassium phosphate (10.2 g, 48.0 mmol) were added sequentially, and the mixture was stirred at 100°C for 3 hours. The reaction solution was cooled to room temperature, diluted with ethyl acetate (200 mL), washed twice with saturated brine (100 mL), and the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: n-hexane / methyl tert-butyl ether = 15 / 1, volume ratio) to give the title compound (6.5 g, yield: 83.0%).

[0309] MS m / z(ESI):655.3[M+H] + .

[0310] Step 7: Preparation of 4-(tert-butoxy)-6-cyclopropyl-2-(ethylsulfonyl)-7-(6-fluoro-5-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)-8-((S)-1-phenylethoxy)quinazoline

[0311] 4-(tert-Butoxy)-6-cyclopropyl-2-(ethylthio)-7-(6-fluoro-5-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)-8-((S)-1-phenylethoxy)quinazoline (6.5 g, 9.9 mmol) was dissolved in dichloromethane (100 mL). Meta-chloroperbenzoic acid (3.4 g, 19.8 mmol) was added portionwise, and the mixture was stirred at 25°C for 2 hours. Saturated aqueous sodium thiosulfate solution (100 mL) was added to the reaction mixture, and the mixture was stirred at 25°C for 0.5 hours. The layers were separated, and the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: n-hexane / ethyl acetate = 3 / 1, volume ratio) to obtain the title compound (2.4 g, yield: 35.2%).

[0312] MS m / z(ESI):687.4[M+H] + .

[0313] Step 8: Preparation of 4-(tert-butoxy)-6-cyclopropyl-2-(ethylsulfonyl)-7-(6-fluoro-5-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)quinazolin-8-ol

[0314] Dissolve 4-(tert-butoxy)-6-cyclopropyl-2-(ethylsulfonyl)-7-(6-fluoro-5-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)-8-((S)-1-phenylethoxy)quinazoline (2.4 g, 3.5 mmol) in anhydrous methanol (50 mL). Add sodium bicarbonate (1.5 g, 17.5 mmol) and 10% palladium on carbon (0.5 g). Replace the mixture with hydrogen three times and stir at 25°C under a hydrogen atmosphere for 3 hours. Filter and concentrate the filtrate under reduced pressure to obtain the title compound (2.0 g, yield: 98.2%).

[0315] MS m / z(ESI):583.3[M+H] + .

[0316] Step 9: Preparation of 4-(tert-butoxy)-6-cyclopropyl-2-(ethylsulfonyl)-8-((4-ethynylbenzyl)oxy)-7-(6-fluoro-5-methyl-1-(tetrahydro-2H-pyran-2)-yl)-1H-indazol-4-yl)quinazoline

[0317] 4-(tert-Butoxy)-6-cyclopropyl-2-(ethylsulfonyl)-7-(6-fluoro-5-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)quinazolin-8-ol (2.0 g, 3.4 mmol) was dissolved in DMF (10 mL). 4-Ethynylbenzyl methanesulfonate (866 mg, 4.1 mmol), potassium iodide (570 mg, 3.4 mmol), and cesium carbonate (2.2 g, 6.8 mmol) were added sequentially, and the mixture was stirred at 60°C for 2 hours. The reaction solution was diluted with ethyl acetate (100 mL) and washed three times with saturated brine (50 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: n-hexane / ethyl acetate = 3 / 1, volume ratio) to obtain the title compound (2.1 g, yield: 87.8%).

[0318] MS m / z(ESI):697.3[M+H] + .

[0319] Step 10: Preparation of 2-((2-oxaspiro[3.3]hept-6-yl)oxy)-4-(tert-butoxy)-6-cyclopropyl-8-((4-ethynylbenzyl)oxy)-7-(6-fluoro-5-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)quinazoline

[0320] 4-(tert-Butoxy)-6-cyclopropyl-2-(ethylsulfonyl)-8-((4-ethynylbenzyl)oxy)-7-(6-fluoro-5-methyl-1-(tetrahydro-2H-pyran-2)-yl)-1H-indazol-4-yl)quinazoline (800 mg, 1.1 mmol) was dissolved in tetrahydrofuran (10 mL). 2-Oxaspiro[3.3]heptan-6-ol (393 mg, 3.3 mmol) and potassium tert-butoxide (258 mg, 2.2 mmol) were added sequentially, and the mixture was stirred at 25°C for 2 hours. The reaction mixture was diluted with ethyl acetate (20 mL) and washed three times with saturated brine (10 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: n-hexane / ethyl acetate = 2 / 1, volume ratio) to obtain the title compound (720 mg, yield: 87.5%).

[0321] MS m / z(ESI):717.3[M+H] + .

[0322] Step 11: Preparation of 2-((2-oxaspiro[3.3]hept-6-yl)oxy)-6-cyclopropyl-8-((4-ethynylbenzyl)oxy)-7-(6-fluoro-5-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)quinazolin-4-ol

[0323] 2-((2-oxaspiro[3.3]hept-6-yl)oxy)-4-(tert-butoxy)-6-cyclopropyl-8-((4-ethynylbenzyl)oxy)-7-(6-fluoro-5-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)quinazoline (720 mg, 1.0 mmol) was dissolved in tetrahydrofuran (10 mL), p-toluenesulfonic acid monohydrate (95 mg, 0.5 mmol) was added, and the mixture was stirred at 25°C for 2 hours. The reaction mixture was diluted with ethyl acetate (20 mL) and washed twice with saturated brine (10 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain the title compound (570 mg, yield: 85.9%).

[0324] MS m / z(ESI):661.4[M+H] + .

[0325] Intermediate Preparation Example 3: Preparation of 6-cyclopropyl-8-((4-ethynylbenzyl)oxy)-7-(6-fluoro-5-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)-2-(7-oxa-2-azaspiro[3.5]nonan-2-yl)quinazolin-4-ol

[0326]

[0327] Step 1: Preparation of 2-(4-(tert-butoxy)-6-cyclopropyl-8-((4-ethynylbenzyl)oxy)-7-(6-fluoro-5-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)quinazolin-2-yl)-7-oxa-2-azaspiro[3.5]nonane

[0328] 4-(tert-Butoxy)-6-cyclopropyl-2-(ethylsulfonyl)-8-((4-ethynylbenzyl)oxy)-7-(6-fluoro-5-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)quinazoline (200 mg, 286 μmol) was dissolved in tetrahydrofuran (5 mL). 7-Oxa-2-azaspiro[3.5]nonane (73 mg, 572 μmol) and potassium tert-butoxide (65 mg, 572 μmol) were added sequentially, and the mixture was stirred at 25°C for 2 hours. The reaction mixture was diluted with ethyl acetate (20 mL) and washed three times with saturated brine (10 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: n-hexane / ethyl acetate = 2 / 1, volume ratio) to obtain the title compound (81 mg, yield: 37.1%).

[0329] MS m / z(ESI):730.5[M+H] + .

[0330] Step 2: Preparation of 6-cyclopropyl-8-((4-ethynylbenzyl)oxy)-7-(6-fluoro-5-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)-2-(7-oxa-2-azaspiro[3.5]non-2-yl)quinazolin-4-ol

[0331] 2-(4-(tert-Butoxy)-6-cyclopropyl-8-((4-ethynylbenzyl)oxy)-7-(6-fluoro-5-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)quinazolin-2-yl)-7-oxa-2-azaspiro[3.5]nonane (81 mg, 103 μmol) was dissolved in tetrahydrofuran (5 mL), p-toluenesulfonic acid monohydrate (20 mg, 105 μmol) was added, and the mixture was stirred at 25°C for 2 hours. The reaction mixture was diluted with ethyl acetate (20 mL) and washed twice with saturated brine (10 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain the title compound (75 mg, yield: 92.6%).

[0332] MS m / z(ESI):674.3[M+H] + .

[0333] Synthesis Example:

[0334] Example 1: Preparation of (2S,4R)-1-((2S)-2-(4-(4-(((2-(2-oxaspiro[3.3]heptane-6-yl)oxy)-6-cyclopropyl-7-((R)-6-fluoro-5-methyl-1H-indazol-4-yl)-4-(methyl(((S)-pyrrolidin-3-yl)amino)quinazolin-8-yl)oxy)methyl)phenyl)-1H-1-1,2,3-triazol-1-yl)-3-methylbutanoyl)-N-((R)-1-[2'-fluoro-[1,1'-biphenyl]-4-yl)-2-hydroxyethyl)-4-hydroxypyrrolidine-2-carboxamide (Compound 1)

[0335]

[0336] Step 1: Preparation of (S)-3-((7-bromo-2-chloro-8-fluoro-6-iodoquinazolin-4-yl)-methyl-amino)pyrrolidine-1-carboxylic acid tert-butyl ester

[0337] 7-Bromo-2,4-dichloro-8-fluoro-6-iodoquinazoline (500 mg, 1.19 mmol) was added to anhydrous tetrahydrofuran (5 mL) and 1.4-dioxane (10 mL) and the temperature was lowered to 0°C. At this temperature, N,N-diisopropylethylamine (460 mg, 3.55 mmol) was added, followed by portionwise addition of (S)-tert-butyl 3-(methylamino)pyrrolidine-1-carboxylate (344 mg, 1.21 mmol). The mixture was stirred at 25°C for 2 hours. Sodium bicarbonate solution (20 mL) was added, and the mixture was extracted three times with ethyl acetate (25 mL). The organic layer was dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 3 / 1, volume ratio) to obtain the title compound (660 mg, yield: 95.1%).

[0338] MS m / z(ESI):585.1[M+H] + .

[0339] Step 2: Preparation of (S)-3-((2-(2-oxaspiro[3.3]heptane-6-yl)oxy)-7-bromo-8-fluoro-6-iodoquinazolin-4-yl)-methyl-amino)pyrrolidine-1-carboxylic acid tert-butyl ester

[0340] (S)-tert-Butyl 3-((7-bromo-2-chloro-8-fluoro-6-iodoquinazolin-4-yl)-methyl-amino)pyrrolidine-1-carboxylate (660 mg, 1.13 mmol), 2-oxaspiro[3.3]heptan-6-ol (180 mg, 1.58 mmol), triethylenediamine (12.7 mg, 0.113 mmol), and cesium carbonate (1.1 g, 3.39 mmol) were added sequentially to acetonitrile (10 mL) and stirred at 60°C for 4 hours. The reaction solution was added dropwise to water (20 mL) and extracted three times with ethyl acetate (15 mL). The organic layer was dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 2 / 1, volume ratio) to obtain the title compound (500 mg, yield: 66.8%).

[0341] MS m / z(ESI):663.2[M+H] + .

[0342] Step 3: Preparation of tert-butyl (S)-3-((2-(2-oxaspiro[3.3]heptane-6-yl)oxy)-8-(benzyloxy)-7-bromo-6-iodoquinazolin-4-yl)-methyl-amino)pyrrolidine-1-carboxylate

[0343] (S)-tert-Butyl 3-((2-(2-oxaspiro[3.3]hept-6-yl)oxy)-7-bromo-8-fluoro-6-iodoquinazolin-4-yl)-methylamino)pyrrolidine-1-carboxylate (500 mg, 754 μmol) was dissolved in tetrahydrofuran (10 mL), followed by benzyl alcohol (122.2 mg, 1.13 mmol) and potassium tert-butoxide (168.3 mg, 1.5 mmol). The mixture was stirred at 50°C for 2 hours. Water (20 mL) was added to the reaction solution, which was then extracted three times with ethyl acetate (10 mL). The organic phases were combined, washed once with saturated brine (10 mL), dried over anhydrous sodium sulfate, and concentrated. The residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 1 / 1, volume ratio) to obtain the title compound (300 mg, 53.0% yield).

[0344] MS m / z(ESI):751.3[M+H] + .

[0345] Step 4: Preparation of (S)-3-((2-(2-oxaspiro[3.3]heptane-6-yl)oxy)-8-(benzyloxy)-7-bromo-6-cyclopropylquinazolin-4-yl)-methyl-amino)pyrrolidine-1-carboxylic acid tert-butyl ester

[0346] Tert-butyl (S)-3-((2-(2-oxaspiro[3.3]heptane-6-yl)oxy)-8-(benzyloxy)-7-bromo-6-iodoquinazolin-4-yl)-methylamino)pyrrolidine-1-carboxylate (300 mg, 399.3 μmol) and cyclopropylboronic acid (206.2 mg, 2.40 mmol) were dissolved in 1,4-dioxane (5 mL) and water (0.5 mL). Potassium phosphate (254.3 mg, 1.20 mmol) and Pd(dppf)Cl2 (32.4 mg, 39.9 μmol) were added. The atmosphere was purged with nitrogen three times and stirred at 80°C for 12 hours. The reaction mixture was concentrated, and the residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 3 / 2, volume ratio) to obtain the title compound (170.5 mg, yield: 64.1%).

[0347] MS m / z(ESI):665.2[M+H] + .

[0348] Step 5: Preparation of tert-butyl (3S)-3-((2-(2-oxaspiro[3.3]heptane-6-yl)oxy)-8-(benzyloxy)-6-cyclopropyl-7-(6-fluoro-5-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)quinazolin-4-yl)-methyl-amino)pyrrolidine-1-carboxylate

[0349] (S)-3-((2-(2-oxaspiro[3.3]heptane-6-yl)oxy)-8-(benzyloxy)-7-bromo-6-cyclopropylquinazolin-4-yl)-methyl-amino)pyrrolidine-1-carboxylic acid tert-butyl ester (170.5 mg, 256.3 μmol) and 6-fluoro-5-methyl-1-(tetrahydro-2H-pyran-2-yl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H- Indazole (184.7 mg, 512.6 μmol) was dissolved in 1,4-dioxane (5 mL) and water (0.5 mL). Potassium phosphate (254.3 mg, 1.20 mmol) was added, and the atmosphere was purged with nitrogen three times. Tris(dibenzylideneacetone)dipalladium (23.4 mg, 25.6 μmol) and 2-dicyclohexylphosphine-2,6-dimethoxy-biphenyl (15.8 mg, 38.4 μmol) were added, and the mixture was stirred at 100°C for 12 hours. The reaction mixture was concentrated, and the residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 1 / 1, volume ratio) to obtain the title compound (153 mg, yield: 73.1%).

[0350] MS m / z(ESI):819.4[M+H] + .

[0351] Step 6: Preparation of tert-butyl (3S)-3-((2-(2-oxaspiro[3.3]heptane-6-yl)oxy)-6-cyclopropyl-7-(6-fluoro-5-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)-8-hydroxyquinazolin-4-yl)-methyl-amino)pyrrolidine-1-carboxylate

[0352] Tert-butyl (3S)-3-((2-(2-oxaspiro[3.3]heptane-6-yl)oxy)-8-(benzyloxy)-6-cyclopropyl-7-(6-fluoro-5-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)quinazolin-4-yl)-methyl-amino)pyrrolidine-1-carboxylate (153 mg, 186.8 μmol) was dissolved in methanol (2 mL). 10% palladium on carbon (20 mg) was added, and hydrogen gas was introduced. The mixture was stirred at 25°C for 2 hours. The reaction mixture was filtered and concentrated to give the title compound (128 mg, 94.1% yield).

[0353] MS m / z(ESI):729.4[M+H] + .

[0354] Step 7: Preparation of tert-butyl (3S)-3-((2-((2-oxaspiro[3.3]heptane-6-yl)oxy)-6-cyclopropyl-8-((4-ethynylbenzyl)oxy)-7-(6-fluoro-5-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)quinazolin-4-yl)-methyl-amino)pyrrolidine-1-carboxylate

[0355] Tert-butyl (3S)-3-((2-(2-oxaspiro[3.3]heptane-6-yl)oxy)-6-cyclopropyl-7-(6-fluoro-5-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)-8-hydroxyquinazolin-4-yl)-methyl-amino)pyrrolidine-1-carboxylate (128 mg, 175.5 μmol) was added to anhydrous DMF (5 mL), followed by the addition of cesium carbonate (143 mg, 438.7 μmol), potassium iodide (29.1 mg, 175.5 μmol) and 4-ethynylbenzyl methanesulfonate (55.3 mg, 263.3 μmol), and the mixture was stirred at 60° C. for 3 hours. Cool to room temperature, dilute with water (20 mL), extract three times with ethyl acetate (25 mL), dry the organic layer over anhydrous sodium sulfate, and concentrate. The residue is purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 2 / 1, volume ratio) to give the title compound (102 mg, yield: 68.9%).

[0356] MS m / z(ESI):843.4[M+H] + .

[0357] Step 8: Preparation of (3S)-3-((2-(2-oxaspiro[3.3]heptane-6-yl)oxy)-6-cyclopropyl-7-(6-fluoro-5-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)-8-(4-(1-((S)-1-((2S,4R)-2-(((R)-1-(2'-fluoro-[1,1'-biphenyl]-4-yl)-2-hydroxyethyl)carbamoyl)-4-hydroxypyrrolidin-1-yl)-3-methyl-1-oxobutan-2-yl)-1,2,3-triazol-4-ylbenzyl)oxy)quinazolin-4-yl)-methyl-amino)pyrrolidine-1-carboxylic acid tert-butyl ester

[0358] (3S)-3-((2-((2-oxaspiro[3.3]heptane-6-yl)oxy)-6-cyclopropyl-8-((4-ethynylbenzyl)oxy)-7-(6-fluoro-5-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)quinazolin-4-yl)-methyl-amino)pyrrolidine-1-carboxylic acid tert-butyl ester (102 mg, 120.9 μmol) and (2S,4R)-1-((S)-2 (R)-1-(2-fluoro-[1,1-biphenyl]-4-yl)-2-hydroxyethyl)-4-hydroxypyrrolidine-2-carboxamide (56.8 mg, 120.9 μmol) was dissolved in DMSO (5 mL) and water (5 mL), and sodium ascorbate (35.9 mg, 181.4 μmol) and copper sulfate pentahydrate (60.4 mg, 241.8 μmol) were added. The mixture was stirred at 25°C for 3 hours. The mixture was diluted with water (20 mL) and extracted three times with ethyl acetate (15 mL). The organic layer was dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel column chromatography (eluent: dichloromethane / methanol = 10 / 1, volume ratio) to obtain the title compound (144 mg, yield: 90.7%).

[0359] MS m / z(ESI):1312.6[M+H] + .

[0360] Step 9: Preparation of (2S,4R)-1-((2S)-2-(4-(4-((((R)-2-(2-oxaspiro[3.3]heptane-6-yl)oxy)-6-cyclopropyl-7-((R)-6-fluoro-5-methyl-1H-indazol-4-yl)-4-(methyl(((S)-pyrrolidin-3-yl)amino)quinazolin-8-yl)oxy)methyl)phenyl)-1H-1-1,2,3-triazol-1-yl)-3-methylbutanoyl)-N-((R)-1-[2'-fluoro-[1,1'-biphenyl]-4-yl)-2-hydroxyethyl)-4-hydroxypyrrolidine-2-carboxamide

[0361] (3S)-3-((2-(2-oxaspiro[3.3]heptane-6-yl)oxy)-6-cyclopropyl-7-(6-fluoro-5-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)-8-(4-(1-((S)-1-((2S,4R)-2-(((R)-1-(2'-fluoro-[1,1'-biphenyl]-4-yl)-2-hydroxy Tert-butyl 1-(2-(2-methyl-1-oxobutan-2-yl)-4-(2-ethyl)amino)-4-hydroxypyrrolidin-1-yl)-3-methyl-1-oxobutan-2-yl)-1,2,3-triazol-4-yl)benzyl)oxy)quinazolin-4-yl)-methyl-amino)pyrrolidine-1-carboxylate (20 mg, 15.24 μmol) was dissolved in dichloromethane (3 mL). Trifluoroacetic acid (0.3 mL) was added under ice-cooling and the mixture was allowed to react at room temperature for 2 hours. The mixture was concentrated, and the crude product was purified by high-performance liquid chromatography to obtain the title compound (2 mg, yield: 11.6%).

[0362] MS m / z(ESI):1128.5[M+H] + .

[0363] 1 H-NMR (400MHz, DMSO-d6) δ13.12(s,1H),8.68(s,1H),8.50(d,J=8.0Hz,1H),7.70(d,J=8.0Hz,2H),7. 54-7.28(m,8H),6.84(d,J=8.0Hz,2H),5.35-5.25(m,2H),5.18(d,J=4.0Hz,1H),5.08-4.82(m,5H),4. 52-4.44(m,4H),4.32(s,1H),3.80-3.59(m,5H),3.25-3.14(m,4H),2.99-2.80(m,4H),2.72-2.67(m,2 H), 2.33-2.09 (m, 5H), 2.00-1.76 (m, 5H), 1.35-1.32 (m, 1H), 1.09 (d, J = 4.0Hz, 3H), 0.72-0.56 (m, 7H).

[0364] Example 2: Preparation of (2S,4R)-1-((2S)-2-(4-(4-(((2-((2-oxaspiro[3.3]hept-6-yl)oxy)-6-cyclopropyl-7-((R)-6-fluoro-5-methyl-1H-indazol-4-yl)-4-(((S)-pyrrolidin-3-yl)amino)quinazolin-8-yl)oxy)methyl)phenyl)-1H-1,2,3-triazol-1-yl)-3-methylbutanoyl)-N-((R)-1-(2'-fluoro-[1,1'-biphenyl]-4-yl)-2-hydroxyethyl)-4-hydroxypyrrolidine-2-carboxamide (Compound 2)

[0365]

[0366] Step 1: Preparation of tert-butyl (3S)-3-((2-((2-oxaspiro[3.3]hept-6-yl)oxy)-6-cyclopropyl-8-((4-ethynylbenzyl)oxy)-7-(6-fluoro-5-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)quinazolin-4-yl)amino)pyrrolidine-1-carboxylate

[0367] 2-((2-oxaspiro[3.3]hept-6-yl)oxy)-6-cyclopropyl-8-((4-ethynylbenzyl)oxy)-7-(6-fluoro-5-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)quinazolin-4-ol (40 mg, 60.0 μmol) was dissolved in tetrahydrofuran (1 mL), and 1H-benzotriazol-1-yloxytripyrrolidinyl hexafluorophosphate (47 mg, 90 μmol) and cesium carbonate (59 mg, 180 μmol) were added. The mixture was stirred at 60°C for 10 minutes. Tert-butyl (S)-3-aminopyrrolidine-1-carboxylate (34 mg, 180 μmol) was added, and the mixture was stirred at 60°C for 2 hours. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: n-hexane / ethyl acetate = 3 / 1) to give the title compound (45 mg, yield: 89.6%).

[0368] MS m / z(ESI):829.4[M+H] + .

[0369] Step 2: Preparation of tert-butyl (3S)-3-((2-((2-oxaspiro[3.3]hept-6-yl)oxy)-6-cyclopropyl-7-((R)-6-fluoro-5-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)-8-((4-(1-((S)-1-((2S,4R)-2-(((R)-1)-(2'-fluoro-[1,1'-biphenyl]-4-yl)-2-hydroxyethyl)carbamoyl)-4-hydroxypyrrolidin-1-yl)-3-methyl-1-oxobutan-2-yl)-1H-1,2,3-triazol-4-yl)benzyl)oxy)quinazolin-4-yl)amino)pyrrolidine-1-carboxylate

[0370] Tert-butyl (3S)-3-((2-((2-oxaspiro[3.3]hept-6-yl)oxy)-6-cyclopropyl-8-((4-ethynylbenzyl)oxy)-7-(6-fluoro-5-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)quinazolin-4-yl)amino)pyrrolidine-1-carboxylate (45 mg, 54 μmol) was dissolved in DMSO (1 mL) and water (0.1 mL). To the mixture was added (2S,4R)-1-((S)-2-azido-3-methylbutanoyl)-N-((R)-1-(2'-fluoro-[1,1'-biphenyl]-4-yl)-2-hydroxyethyl)-4-hydroxypyrrolidine-2-carboxamide (31 mg, 65 μmol), sodium ascorbate (22 mg, 108 μmol), and copper sulfate pentahydrate (27 mg, 108 μmol), followed by stirring at 0°C for 2 hours. The reaction mixture was warmed to room temperature and purified by reverse-phase column chromatography (eluent: pure water / acetonitrile = 9 / 1, volume ratio) to obtain an axial chiral mixture. This mixture was then purified by high-performance liquid chromatography to obtain the title compound (15 mg, yield: 21.0%).

[0371] MS m / z(ESI):1298.7[M+H] + .

[0372] Step 3: Preparation of (2S,4R)-1-((2S)-2-(4-(4-(((2-((2-oxaspiro[3.3]hept-6-yl)oxy)-6-cyclopropyl-7-((R)-6-fluoro-5-methyl-1H-indazol-4-yl)-4-(((S)-pyrrolidin-3-yl)amino)quinazolin-8-yl)oxy)methyl)phenyl)-1H-1,2,3-triazol-1-yl)-3-methylbutanoyl)-N-((R)-1-(2'-fluoro-[1,1'-biphenyl]-4-yl)-2-hydroxyethyl)-4-hydroxypyrrolidine-2-carboxamide

[0373] (3S)-3-((2-((2-oxaspiro[3.3]hept-6-yl)oxy)-6-cyclopropyl-7-(6-fluoro-5-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)-8-((4-(1-((S)-1-((2S,4R)-2-(((R)-1)-(2'-fluoro-[1,1'-biphenyl]-4-yl)- Tert-butyl 2-(2-hydroxyethyl)carbamoyl)-4-hydroxypyrrolidin-1-yl)-3-methyl-1-oxobutan-2-yl)-1H-1,2,3-triazol-4-yl)benzyl)oxy)quinazolin-4-yl)amino)pyrrolidine-1-carboxylate (15 mg, 11 μmol) was dissolved in dichloromethane (1 mL), and trifluoroacetic acid (0.1 mL) was added. The mixture was stirred at 25°C for 3 hours. The reaction mixture was adjusted to pH = 8 with triethylamine and concentrated under reduced pressure. The residue was purified by high-performance liquid chromatography to obtain the title compound (1.2 mg, yield: 4.6%).

[0374] MS m / z(ESI):1114.5[M+H] + .

[0375] 1 H-NMR (400MHz, DMSO-d6) δ13.13(s,1H),8.68(s,1H),8.51(d,J=8.0Hz,1H),8.20(d,J=4.0Hz,1H),7.71(d,J=8.0Hz,1H),7.64(s,1H),7.5 3-7.48(m,3H),7.42-7.37(m,3H),7.33-7.28(m,2H),6.84(d,J=8.0Hz,2H),5.35(d,J=12.0Hz,1H),5.28(d,J=12.0Hz,1H),5.18(d,J=4.0H z,1H),5.06(t,J=4.0Hz,1H),4.89-4.75(m,3H),4.52-4.44(m,4H),4.32(s,1H),3.79-3.53(m,4H),3.34-3.18(m,6H),2.71-2.67(m,2H),2 .33-2.27(m,3H),2.11-2.01(m,3H),1.98(d,J=2.0Hz,3H),1.82-1.77(m,2H),1.45-1.31(m,2H),1.09(d,J=4.0Hz,3H),0.85-0.57(m,7H).

[0376] Example 3: Preparation of (2S,4R)-1-((2S)-2-(4-(4-(((2-((2-oxaspiro[3.3]hept-6-yl)oxy)-6-cyclopropyl-7-((R)-6-fluoro-5-methyl-1H-indazol-4-yl)-4-(methyl((R)-pyrrolidin-3-yl)amino)quinazolin-8-yl)oxy)methyl)phenyl)-1H-1,2,3-triazol-1-yl)-3-methylbutanoyl)-N-((R)-1-(2'-fluoro-[1,1'-biphenyl]-4-yl)-2-hydroxyethyl)-4-hydroxypyrrolidine-2-carboxamide (Compound 4)

[0377]

[0378] Step 1: Preparation of tert-butyl (3R)-3-((2-((2-oxaspiro[3.3]hept-6-yl)oxy)-6-cyclopropyl-8-((4-ethynylbenzyl)oxy)-7-(6-fluoro-5-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)quinazolin-4-yl)(methyl)amino)pyrrolidine-1-carboxylate

[0379] 2-((2-oxaspiro[3.3]hept-6-yl)oxy)-6-cyclopropyl-8-((4-ethynylbenzyl)oxy)-7-(6-fluoro-5-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)quinazolin-4-ol (50 mg, 75.8 μmol) was dissolved in tetrahydrofuran (1 mL), 1H-benzotriazol-1-yloxytripyrrolidinyl hexafluorophosphate (50.2 mg, 114 μmol) and cesium carbonate (74 mg, 227.4 μmol) were added, and the mixture was stirred at 60°C for 10 minutes. Tert-butyl (R)-3-(methylamino)pyrrolidine-1-carboxylate (45.4 mg, 227.4 μmol) was added, and the mixture was stirred at 60°C for 2 hours. The reaction solution was cooled to room temperature and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: n-hexane / ethyl acetate = 3 / 1, volume ratio) to give the title compound (60 mg, yield: 94.0%).

[0380] MS m / z(ESI):843.5[M+H] + .

[0381] Step 2: Preparation of tert-butyl (3R)-3-((2-((2-oxaspiro[3.3]hept-6-yl)oxy)-6-cyclopropyl-7-((R)-6-fluoro-5-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)-8-((4-(1-((S)-1-((2S,4R)-2-(((R)-1)-(2'-fluoro-[1,1'-biphenyl]-4-yl)-2-hydroxyethyl)carbamoyl)-4-hydroxypyrrolidin-1-yl)-3-methyl-1-oxobutan-2-yl)-1H-1,2,3-triazol-4-yl)benzyl)oxy)quinazolin-4-yl)(methyl)amino)pyrrolidine-1-carboxylate

[0382] Tert-butyl (3R)-3-((2-((2-oxaspiro[3.3]hept-6-yl)oxy)-6-cyclopropyl-8-((4-ethynylbenzyl)oxy)-7-(6-fluoro-5-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)quinazolin-4-yl)(methyl)amino)pyrrolidine-1-carboxylate (60 mg, 71 μmol) was dissolved in DMSO (1 mL) and water (0.1 mL). To a solution of 4-nitro-1,4-dihydro-1,4-dihydro-2-pyrrolidine-2-carboxamide (L) was added (2S,4R)-1-((S)-2-azido-3-methylbutanoyl)-N-((R)-1-(2'-fluoro-[1,1'-biphenyl]-4-yl)-2-hydroxyethyl)-4-hydroxypyrrolidine-2-carboxamide (33 mg, 71 μmol), sodium ascorbate (22 mg, 107 μmol), and copper sulfate pentahydrate (27 mg, 107 μmol), followed by stirring at 0°C for 2 hours. The reaction solution was warmed to room temperature and purified by reverse-phase column chromatography (eluent: pure water / acetonitrile = 9 / 1, volume ratio) to obtain an axial chiral mixture. This mixture was then purified by high performance liquid chromatography to obtain the title compound (20 mg, yield: 21.4%).

[0383] MS m / z(ESI):1312.6[M+H] + .

[0384] Step 3: Preparation of (2S,4R)-1-((2S)-2-(4-(4-(((2-((2-oxaspiro[3.3]hept-6-yl)oxy)-6-cyclopropyl-7-((R)-6-fluoro-5-methyl-1H-indazol-4-yl)-4-(methyl((R)-pyrrolidin-3-yl)amino)quinazolin-8-yl)oxy)methyl)phenyl)-1H-1,2,3-triazol-1-yl)-3-methylbutanoyl)-N-((R)-1-(2'-fluoro-[1,1'-biphenyl]-4-yl)-2-hydroxyethyl)-4-hydroxypyrrolidine-2-carboxamide

[0385] (3R)-3-((2-((2-oxaspiro[3.3]hept-6-yl)oxy)-6-cyclopropyl-7-((R)-6-fluoro-5-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)-8-((4-(1-((S)-1-((2S,4R)-2-(((R)-1)-(2'-fluoro-[1,1'-biphenyl]-4-yl)- Tert-butyl (2-hydroxyethyl)carbamoyl)-4-hydroxypyrrolidin-1-yl)-3-methyl-1-oxobutan-2-yl)-1H-1,2,3-triazol-4-yl)benzyl)oxy)quinazolin-4-yl)(methyl)amino)pyrrolidine-1-carboxylate (20 mg, 11 μmol) was dissolved in dichloromethane (1 mL), and trifluoroacetic acid (0.1 mL) was added. The mixture was stirred at 25°C for 3 hours. The reaction mixture was adjusted to pH 8 with triethylamine, concentrated under reduced pressure, and the residue was purified by high-performance liquid chromatography to obtain the title compound (3.3 mg, yield: 19.2%).

[0386] MS m / z(ESI):1128.5[M+H] + .

[0387] 1 H-NMR(400MHz,DMSO-d6)δ13.12(s,1H),8.68(s,1H),8.51(d,J=8.0Hz,1H ),7.71(d,J=8.0Hz,2H),7.52-7.48(m,3H),7.44-7.37(m,4H),7.33-7.28 (m,2H),6.85(d,J=8.0Hz,2H),5.35(d,J=12.0Hz,1H),5.27(d,J=12.0Hz, 1H),5.18(d,J=4.0Hz,1H),5.05(t,J=8.0Hz,1H),4.93-4.82(m,3H),4.52 -4.44(m,5H),4.32(s,1H),3.81-3.59(m,4H),3.34(s,3H),3.25-3.14(m, 1H),3.00-2.85(m,3H),2.72-2.68(m,2H),2.33-2.30(m,2H),2.11-2.06( m,3H),2.00(d,J=2.0Hz,3H),1.83-1.72(m,3H),1.38-1.35(m,1H),1.26- 1.22(m,2H),1.09(d,J=4.0Hz,3H),0.72-0.64(m,1H),0.58-0.50(m,6H).

[0388] Example 4: Preparation of (2S,4R)-1-((2S)-2-(4-(4-(((2-((2-oxaspiro[3.3]hept-6-yl)oxy)-6-cyclopropyl-7-((R)-6-fluoro-5-methyl-1H-indazol-4-yl)-4-(((S)-pyrrolidin-3-yl)oxy)quinazolin-8-yl)oxy)methyl)phenyl)-1H-1,2,3-triazol-1-yl)-3-methylbutanoyl)-N-((R)-1-(2'-fluoro-[1,1'-biphenyl]-4-yl)-2-hydroxyethyl)-4-hydroxypyrrolidine-2-carboxamide (Compound 3)

[0389]

[0390] Step 1: Preparation of tert-butyl (3S)-3-((2-((2-oxaspiro[3.3]hept-6-yl)oxy)-6-cyclopropyl-8-((4-ethynylbenzyl)oxy)-7-(6-fluoro-5-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)quinazolin-4-yl)oxy)pyrrolidine-1-carboxylate

[0391] 2-((2-oxaspiro[3.3]hept-6-yl)oxy)-6-cyclopropyl-8-((4-ethynylbenzyl)oxy)-7-(6-fluoro-5-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)quinazolin-4-ol (50 mg, 75.6 μmol) was dissolved in DMF (1 mL), and tert-butyl (R)-3-((methylsulfonyl)oxy)pyrrolidine-1-carboxylate (60 mg, 225.8 μmol), cesium carbonate (74 mg, 225.8 μmol) and potassium iodide (13 mg, 75.6 μmol) were added sequentially, and the mixture was stirred at 80°C for 20 hours. The reaction solution was cooled to room temperature, the solid was filtered off, and the filtrate was purified by reverse column chromatography (eluent: pure water / acetonitrile = 15 / 1, volume ratio) to obtain the title compound (60 mg, yield: 95.5%)

[0392] MS m / z(ESI):830.7[M+H] + .

[0393] Step 2: Preparation of tert-butyl (3S)-3-((2-((2-oxaspiro[3.3]hept-6-yl)oxy)-6-cyclopropyl-7-((R)-6-fluoro-5-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)-8-((4-(1-((S)-1-((2S,4R)-2-(((R)-1)-(2'-fluoro-[1,1'-biphenyl]-4-yl)-2-hydroxyethyl)carbamoyl)-4-hydroxypyrrolidin-1-yl)-3-methyl-1-oxobutan-2-yl)-1H-1,2,3-triazol-4-yl)benzyl)oxy)quinazolin-4-yl)oxy)pyrrolidine-1-carboxylate

[0394] Tert-butyl (3S)-3-((2-((2-oxaspiro[3.3]hept-6-yl)oxy)-6-cyclopropyl-8-((4-ethynylbenzyl)oxy)-7-(6-fluoro-5-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)quinazolin-4-yl)oxy)pyrrolidine-1-carboxylate (60 mg, 72.2 μmol) was dissolved in DMSO (1 mL) and water (0.1 mL). (2S,4R)-1-((S)-2-azido-3-methylbutanoyl)-N-((R)-1-(2'-fluoro-[1,1'-biphenyl]-4-yl)-2-hydroxyethyl)-4-hydroxypyrrolidine-2-carboxamide (40 mg, 86.6 μmol), sodium ascorbate (28 mg, 144.4 μmol), and copper sulfate pentahydrate (36 mg, 144.4 μmol) were added sequentially, and the mixture was stirred at 0°C for 2 hours. The reaction solution was warmed to room temperature and purified by reverse-phase column chromatography (eluent: pure water / acetonitrile = 9 / 1, volume ratio) to obtain an axial chiral mixture. This mixture was further purified by high-performance liquid chromatography to obtain the title compound (25 mg, yield: 26.6%).

[0395] MS m / z(ESI):1299.7[M+H] + .

[0396] Step 3: Preparation of (2S,4R)-1-((2S)-2-(4-(4-(((2-((2-oxaspiro[3.3]hept-6-yl)oxy)-6-cyclopropyl-7-((R)-6-fluoro-5-methyl-1H-indazol-4-yl)-4-(((S)-pyrrolidin-3-yl)oxy)quinazolin-8-yl)oxy)methyl)phenyl)-1H-1,2,3-triazol-1-yl)-3-methylbutanoyl)-N-((R)-1-(2'-fluoro-[1,1'-biphenyl]-4-yl)-2-hydroxyethyl)-4-hydroxypyrrolidine-2-carboxamide

[0397] (3S)-3-((2-((2-oxaspiro[3.3]hept-6-yl)oxy)-6-cyclopropyl-7-((R)-6-fluoro-5-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)-8-((4-(1-((S)-1-((2S,4R)-2-(((R)-1)-(2'-fluoro-[1,1'-biphenyl]-4-yl) Tert-butyl 1-hydroxy-2-methyl-1-oxobutan-2-yl-1H-1,2,3-triazol-4-yl)-2-hydroxyethyl)carbamoyl)-4-hydroxypyrrolidin-1-yl)-3-methyl-1-oxobutan-2-yl)-1H-1,2,3-triazol-4-yl)benzyl)oxy)quinazolin-4-yl)oxy)pyrrolidine-1-carboxylate (25 mg, 19.2 μmol) was dissolved in dichloromethane (1 mL), and trifluoroacetic acid (0.1 mL) was added. The mixture was stirred at 25°C for 3 hours. The reaction mixture was adjusted to pH = 8 with triethylamine, concentrated under reduced pressure, and purified by high-performance liquid chromatography to obtain the title compound (4.7 mg, yield: 21.9%).

[0398] MS m / z(ESI):1115.4[M+H] + .

[0399] 1 H-NMR (400MHz, DMSO-d6) δ13.15(s,1H),8.68(s,1H),8.51(d,J=4.0Hz,1H),7.72(d,J=8.0,1H),7.53-7.47(m,4H),7 .43-7.37(m,4H),7.33-7.28(m,2H),6.85(t,J=8.0Hz,2H),5.35(d,J=8.0Hz,1H),5.18(d,J=4.0Hz,1H),4.89-4.84( m,3H),4.54-4.51(m,3H),4.48-4.44(m,2H),4.32(s,1H),3.81-3.59(m,6H),3.09-2.80(m,5H),2.74-2.67(m,2H),2 .41-2.33(m,3H),2.11-1.96(m,6H),1.83-1.75(m,2H),1.36-1.30(m,3H),1.09(d,J=4.0Hz,3H),0.72-0.56(m,7H).

[0400] Example 5: Preparation of (2S,4R)-1-((2S)-2-(4-(4-(((2-((2-oxaspiro[3.3]hept-6-yl)oxy)-6-cyclopropyl-7-((R)-6-fluoro-5-methyl-1H-indazol-4-yl)-4-(((R)-pyrrolidin-3-yl)oxy)quinazolin-8-yl)oxy)methyl)phenyl)-1H-1,2,3-triazol-1-yl)-3-methylbutanoyl)-N-((R)-1-(2'-fluoro-[1,1'-biphenyl]-4-yl)-2-hydroxyethyl)-4-hydroxypyrrolidine-2-carboxamide (Compound 5)

[0401]

[0402] Using the synthetic route of Example 4, the first step reaction raw material (R)-3-((methylsulfonyl)oxy)pyrrolidine-1-carboxylic acid tert-butyl ester was replaced with (S)-3-((methylsulfonyl)oxy)pyrrolidine-1-carboxylic acid tert-butyl ester to obtain the title compound (2.8 mg, yield: 8.5%).

[0403] MS m / z(ESI):1115.4[M+H] + .

[0404] 1 H-NMR (400MHz, DMSO-d6) δ13.14(s,1H),8.68(s,1H),8.51(d,J=8.0Hz,1H),7.72(d,J=8.0,1H),7.53-7.47(m,4H),7.43- 7.37(m,4H),7.33-7.28(m,2H),6.85(t,J=8.0Hz,2H),5.35(d,J=8.0Hz,1H),5.18(d,J=4.0Hz,1H),4.89-4.84(m,2H),4. 54-4.44(m,4H),4.32(s,1H),3.80-3.59(m,6H),3.14-3.01(m,4H),2.89-2.83(m,2H),2.74-2.67(m,2H),2.43-2.33(m,3 H), 2.15-2.06 (m, 2H), 2.01-1.97 (m, 4H), 1.83-1.76 (m, 1H), 1.35-1.31 (m, 2H), 1.09 (d, J = 4.0Hz, 3H), 0.72-0.56 (m, 7H).

[0405] Example 6: Preparation of (2S,4R)-1-((2S)-2-(4-(4-(2-(2-(2-oxospiro[3.3]heptane-6-yl)oxy)-6-cyclopropyl-4-(ethyl((R)-pyrrolidin-3-yl)amino)-7-((R)-6-fluoro-5-methyl-1-indol-4-yl)quinazolin-8-yl)methyl)phenyl)-1H-1,2,3-triazol-1-yl)-3-methylbutanyl)-N-((R)-1-(2-fluoro-[1,1-biphenyl]-4-yl)-2-hydroxyethyl)-4-hydroxypyrrolidine-2-carboxamide (Compound 12)

[0406]

[0407] Using the synthetic route in Example 1, the first step reaction raw material (S)-3-(methylamino)pyrrolidine-1-carboxylic acid tert-butyl ester was replaced with (R)-3-(ethylamino)pyrrolidine-1-carboxylic acid tert-butyl ester to obtain the title compound (5 mg, yield: 6.5%).

[0408] MS m / z(ESI):1142.5[M+H] + .

[0409] 1H-NMR (400MHz, DMSO-d6) δ13.13(s,1H),8.68(s,1H),8.51(d,J=8.0Hz,1H),7.70(d ,J=8.0Hz,2H),7.55(s,1H),7.50(dd,J=8.0,5.6Hz,3H),7.43-7.35(m,4H),7.33-7 .27(m,3H),6.84(d,J=8.0Hz,2H),5.34(d,J=10.4Hz,1H),5.30-5.21(m,2H),5.18( d,J=4.0Hz,1H),5.12-5.01(m,1H),4.91-4.81(m,3H),4.73(d,J=7.6Hz,1H),4.56-4 .43(m,4H),4.32(s,1H),3.79(dd,J=11.2,4.4Hz,1H),3.73-3.56(m,5H),3.16-2.9 8(m,2H),2.96-2.75(m,2H),2.74-2.65(m,2H),2.37-2.26(m,3H),2.19-2.03(m,2H) ,2.01(d,J=2.4Hz,3H),1.80(dd,J=8.4,4.4Hz,1H),1.38(t,J=8.0Hz,1H),1.27-1. 17 (m, 5H), 1.09 (d, J = 6.4Hz, 3H), 0.72 (dd, J = 6.8, 3.2Hz, 4H), 0.63 (t, J = 4.8Hz, 3H).

[0410] Example 7: Preparation of (2S,4R)-1-((2S)-2-(4-(4-(2-(2-(2-oxospiro[3.3]heptane-6-yl)oxy)-6-cyclopropyl-4-(ethyl((S)-pyrrolidin-3-yl)amino)-7-((R)-6-fluoro-5-methyl-1-indol-4-yl)quinazolin-8-yl)methyl)phenyl)-1H-1,2,3-triazol-1-yl)-3-methylbutanyl)-N-((R)-1-(2-fluoro-[1,1-biphenyl]-4-yl)-2-hydroxyethyl)-4-hydroxypyrrolidine-2-carboxamide (Compound 11)

[0411]

[0412] Using the synthetic route in Example 1, the first step reaction raw material (S)-3-(methylamino)pyrrolidine-1-carboxylic acid tert-butyl ester was replaced with (S)-3-(ethylamino)pyrrolidine-1-carboxylic acid tert-butyl ester to obtain the title compound (4 mg, yield: 9.3%).

[0413] MS m / z(ESI):1142.5[M+H]+ .

[0414] 1 H-NMR (400MHz, DMSO-d6) δ13.13(s,1H),8.68(s,1H),8.51(d,J=8.0Hz,1H),7.69( d,J=7.2Hz,2H),7.59-7.46(m,3H),7.43-7.37(m,4H),7.34-7.28(m,3H),6.87-6.7 9(m,2H),5.38-5.21(m,3H),5.18(d,J=3.6Hz,1H),5.05(t,J=6.0Hz,1H),4.93-4. 80(m,3H),4.73(d,J=7.2Hz,1H),4.56-4.43(m,4H),4.32(s,1H),3.79(dd,J=11.2, 4.4Hz,1H),3.74-3.56(m,5H),3.14-3.04(m,1H),3.00(s,1H),2.93-2.84(m,1H), 2.84-2.75(m,1H),2.75-2.65(m,2H),2.36-2.25(m,3H),2.15-2.03(m,2H),2.01(d ,J=2.4Hz,3H),1.96-1.85(m,1H),1.85-1.73(m,1H),1.38(t,J=8.0Hz,1H),1.28- 1.17(m,5H),1.09(d,J=6.4Hz,3H),0.72(dd,J=6.8,3.2Hz,4H),0.66-0.47(m,3H).

[0415] Example 8: Preparation of (2S,4R)-1-((2S)-2-(4-(((6-cyclopropyl-7-((R)-6-fluoro-5-methyl-1H-indazol-4-yl)-4-(methyl((R)-pyrrolidin-3-yl)amino)-2-(2-oxa-7-azaspiro[3.5]nonan-7-yl)quinazolin-8-yl)oxy)methyl)phenyl)-1H-1,2,3-triazol-1-yl)-3-methylbutanoyl)-N-((R)-1-(2'-fluoro-[1,1'-biphenyl]-4-yl)-2-hydroxyethyl)-4-hydroxypyrrolidine-2-carboxamide (Compound 7)

[0416]

[0417] Using the synthetic route of Example 1, the first step reaction raw material (S)-3-(methylamino)pyrrolidine-1-carboxylic acid tert-butyl ester was replaced with (R)-3-(methylamino)pyrrolidine-1-carboxylic acid tert-butyl ester; the second step reaction raw material 2-oxaspiro[3.3]heptane-6-ol was replaced with 2-oxo-7-azaspiro[3.5]nonane to obtain the title compound (1.12 mg, yield: 6.5%).

[0418] MS m / z(ESI):1141.5[M+H] + .

[0419] 1 H-NMR (400MHz, DMSO-d6) δ13.09(s,1H),8.67(s,1H),8.57-8.47(m,1H),7.67(dd,J=8.4,6.4Hz,2H),7.56-7.45(m,4H),7.43-7.35(m,4H), 7.31(d,J=7.2Hz,2H),7.25(d,J=4.0Hz,1H),6.83-6.74(m,2H),5.34(d,J=10.4Hz,1H),5.26(d,J=11.2Hz,2H),5.18(d,J=3.6Hz,1H),4.93 -4.72(m,4H),4.46(t,J=8.0Hz,1H),4.36-4.25(m,5H),3.86-3.67(m,6H),3.62(t,J=6.0Hz,2H),3.18(d,J=7.2Hz,3H),3.00-2.77(m,2H), 2.28-2.04(m,3H),2.00(t,J=2.4Hz,3H),1.94-1.74(m,6H),1.39-1.2 6(m,1H),1.09(d,J=6.4Hz,3H),0.78-0.69(m,3H),0.68-0.45(m,5H).

[0420] Example 9: Preparation of (2S,4R)-1-((2S)-2-(4-(4-(((6-cyclopropyl-7-((R)-6-fluoro-5-methyl-1H-indazol-4-yl))-4-(methyl((R)-pyrrolidin-3-yl)amino)-2-(7-oxa-2-azaspiro[3.5]nonan-2-yl)quinazolin-8-yl)oxy)methyl)phenyl)-1H-1,2,3-triazol-1-yl)-3-methylbutanoyl)-N-((R)-1-(2'-fluoro-[1,1'-biphenyl]-4-yl)-2-hydroxyethyl)-4-hydroxypyrrolidine-2-carboxamide (Compound 9)

[0421]

[0422] Step 1: Preparation of tert-butyl (3R)-3-((6-cyclopropyl-8-((4-ethynylbenzyl)oxy)-7-(6-fluoro-5-methyl-1-(tetrahydro-2H-pyran-2-yl))-1H-indazol-4-yl)-2-(7-oxa-2-azaspiro[3.5]nonan-2-yl)quinazolin-4-yl)(methyl)amino)pyrrolidine-1-carboxylate

[0423] 6-Cyclopropyl-8-((4-ethynylbenzyl)oxy)-7-(6-fluoro-5-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)-2-(7-oxa-2-azaspiro[3.5]nonan-2-yl)quinazolin-4-ol (35 mg, 44.15 μmol) and (R)-tert-butyl 3-(methylamino)pyrrolidine-1-carboxylate (26.8 mg, 132.45 μmol) were dissolved in tetrahydrofuran (2 mL), and 1H-benzotriazol-1-yloxytripyrrolidinyl hexafluorophosphate (46.4 mg, 88.3 μmol) and cesium carbonate (43.6 mg, 132.45 μmol) were added. The mixture was stirred at 60°C for 2 hours. Ethyl acetate (10 mL) was added to the reaction solution, and the mixture was filtered. The filtrate was spin-dried to give the title compound (31 mg, yield: 82.1%).

[0424] MS m / z(ESI):856.4[M+H] + .

[0425] Step 2: Preparation of tert-butyl (3R)-3-((6-cyclopropyl-7-(6-fluoro-5-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)-8-((4-(1-((S)-1-((2S,4R)-2-(((R)-1-(2'-fluoro-[1,1'-biphenyl]-4-yl))-2-hydroxyethyl)carbamoyl)-4-hydroxypyrrolidin-1-yl)-3-methyl-1-oxobutan-2-yl)-1H-1,2,3-triazol-4-yl)benzyl)oxy)-2-(7-oxa-2-azaspiro[3.5]nonan-2-yl)quinazolin-4-yl)(methyl)amino)pyrrolidine-1-carboxylate

[0426] Tert-butyl (3R)-3-((6-cyclopropyl-8-((4-ethynylbenzyl)oxy)-7-(6-fluoro-5-methyl-1-(tetrahydro-2H-pyran-2-yl))-1H-indazol-4-yl)-2-(7-oxa-2-azaspiro[3.5]nonan-2-yl)quinazolin-4-yl)(methyl)amino)pyrrolidine-1-carboxylate (31 mg, 36.2 μmol) and (2S,4R)-1-((S)-2-azide ((R)-1-(2'-fluoro-[1,1'-biphenyl]-4-yl)-2-hydroxyethyl)-4-hydroxypyrrolidine-2-carboxamide (18 mg, 36.2 μmol) was dissolved in DMSO (2 mL) and water (0.2 mL), followed by the addition of sodium ascorbate (11.3 mg, 54.3 μmol) and anhydrous copper sulfate (19 mg, 72.4 μmol). The mixture was stirred at 25°C for 2 hours. Water (5 mL) was added to the reaction solution, which was extracted three times with ethyl acetate (5 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by reverse-phase column chromatography (eluent: pure water / acetonitrile = 9 / 1, volume ratio) to obtain an axial chiral mixture. This mixture was then purified by high-performance liquid chromatography to afford the title compound (41.4 mg, yield: 86.2%).

[0427] MS m / z(ESI):1325.6[M+H] + .

[0428] Step 3: Preparation of (2S,4R)-1-((2S)-2-(4-(4-(((6-cyclopropyl-7-((R)-6-fluoro-5-methyl-1H-indazol-4-yl))-4-(methyl((R)-pyrrolidin-3-yl)amino)-2-(7-oxa-2-azaspiro[3.5]nonan-2-yl)quinazolin-8-yl)oxy)methyl)phenyl)-1H-1,2,3-triazol-1-yl)-3-methylbutanoyl)-N-((R)-1-(2'-fluoro-[1,1'-biphenyl]-4-yl)-2-hydroxyethyl)-4-hydroxypyrrolidine-2-carboxamide

[0429] (3R)-3-((6-cyclopropyl-7-(6-fluoro-5-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)-8-((4-(1-((S)-1-((2S,4R)-2-(((R)-1-(2'-fluoro-[1,1'-biphenyl]-4-yl))-2-hydroxyethyl)carbamoyl)-4-hydroxypyrrolidin-1-yl)-3- Tert-butyl ((methyl)-1-oxobutan-2-yl)-1H-1,2,3-triazol-4-yl)benzyl)oxy)-2-(7-oxa-2-azaspiro[3.5]nonan-2-yl)quinazolin-4-yl)(methyl)amino)pyrrolidine-1-carboxylate (8 mg, 5.73 μmol) was dissolved in dichloromethane (2 mL), trifluoroacetic acid (400 μL) was added, and the mixture was stirred at 25°C for 2 hours. The reaction solution was concentrated, and the residue was diluted with ethyl acetate (5 mL). The pH was adjusted to 8-9 with sodium bicarbonate solution. The layers were separated, and the organic phase was dried over anhydrous sodium sulfate and concentrated. The crude product was purified by high-performance liquid chromatography to obtain the title compound (2.5 mg, 36.3% yield).

[0430] MS m / z(ESI):1141.5[M+H] + .

[0431] 1 H-NMR (400MHz, CD3OD) δ8.43(d,J=1.6Hz,1H),7.56-7.15(m,13H),6.83(d,J=8.4Hz,2H),5.36(d ,J=10.4Hz,1H),5.25-5.02(m,4H),4.90-4.81(m,2H),4.65-4.48(m,4H),4.03-3.80(m,8H),3.7 1-3.55(m,5H),3.42-3.35(m,2H),2.71-2.47(m,3H),2.44-2.21(m,3H),2.11-1.98(m,4H),1.89 -1.82(m,3H),1.45-1.41(m,2H),1.17(d,J=6.4Hz,3H),0.83(d,J=6.4Hz,3H),0.61-0.58(m,2H).

[0432] Example 10: Preparation of (2S,4R)-1-((2S)-2-(4-(4-(((6-cyclopropyl-7-((R)-6-fluoro-5-methyl-1H-indazol-4-yl))-4-(((R)-pyrrolidin-3-yl)oxy)-2-(7-oxa-2-azaspiro[3.5]nonan-2-yl)quinazolin-8-yl)oxy)methyl)phenyl)-1H-1,2,3-triazol-1-yl)-3-methylbutanoyl)-N-((R)-1-(2'-fluoro-[1,1'-biphenyl]-4-yl)-2-hydroxyethyl)-4-hydroxypyrrolidine-2-carboxamide (Compound 8)

[0433]

[0434] The synthetic route in Example 4 was adopted, and the first step reaction raw material 2-((2-oxaspiro[3.3]hept-6-yl)oxy)-6-cyclopropyl-8-((4-ethynylbenzyl)oxy)-7-(6-fluoro-5-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)quinazolin-4-ol was replaced with 6-cyclopropyl-8-((4-ethynylbenzyl)oxy)-7-(6-fluoro-5-methyl- The reaction mixture was purified by mixing the raw material (R)-tert-butyl 3-((methylsulfonyl)oxy)pyrrolidine-1-carboxylate with tert-butyl 3-((methylsulfonyl)oxy)pyrrolidine-1-carboxylate to give the title compound (3 mg, yield: 11.5%).

[0435] MS m / z(ESI):1128.4[M+H] + .

[0436] 1H-NMR(400MHz,DMSO-d6)δ13.10(s,1H),8.66(s,1H),8.51(d,J=8.0Hz,1H ),7.66(d,J=8.0Hz,2H),7.54-7.47(m,4H),7.44-7.38(m,4H),7.34-7.28 (m,3H),6.84(d,J=8.0Hz,2H),5.65-5.55(m,1H),5.33(t,J=8.0Hz,2H),5 .19(d,J=4.0Hz,1H),4.89-4.83(m,3H),4.46(t,J=8.0Hz,1H),4.33-4.29 (m,1H),3.90(s,3H),3.80-3.77(m,1H),3.72(d,J=12.0Hz,1H),3.66-3.5 6(m,6H),3.21-3.18(m,2H),3.04-3.01(m,2H),2.89-2.81(m,2H),2.17-2 .08(m,3H),2.01-1.96(m,4H),1.82-1.75(m,4H),1.33-1.29(m,2H),1.09 (d,J=8.0Hz,3H),0.78-0.69(m,3H),0.64-0.59(m,3H),0.51-0.47(m,1H).

[0437] Biological Examples:

[0438] Test Example 1: In vitro enzymatic ternary complex binding activity test of the compound with KRAS-G12D (guanine-5'-triphosphate (GTP) hydrolase G12D mutant) / VHL

[0439] Test system and parameters

[0440] protein:

[0441] Human KRAS / K-Ras (G12D & Q61H) protein (His tag, Sino Biological): 300 nM

[0442] Human VHL recombinant protein (Abnova): 300nM

[0443] Label:

[0444] Anti-6xHis AlphaLISA acceptor beads (PerkinElmer): 20 μg / mL

[0445] α-Glutathione donor microspheres (PerkinElmer): 20 μg / mL

[0446] Buffer: AlphaLISA 10X buffer (PerkinElmer)

[0447] Microplate reader: BMG PHERAstar Fluorescence, AlphaLISA method, excitation wavelength 680nm, emission wavelength 615nm

[0448] Test steps

[0449] Preincubate the test compound with KRAS / K-Ras (G12D & Q61H) protein and recombinant human VHL protein at 23°C for 60 minutes to allow binding. Add acceptor and donor microspheres and incubate at 23°C in the dark for 60 minutes. Place the reaction plate in a microplate reader, and read the signal in each well using the AlphaLISA assay.

[0450] Data processing

[0451] Wells containing samples (containing different dilutions of sample, KRAS / K-Ras (G12D & Q61H) protein, recombinant human VHL protein, anti-6xHis AlphaLISA acceptor beads, and α-glutathione donor microspheres) served as test groups, while the vehicle group (containing KRAS / K-Ras (G12D & Q61H) protein, recombinant human VHL protein, anti-6xHis AlphaLISA acceptor beads, and α-glutathione donor microspheres in 1% DMSO) served as negative controls. The maximum excitation signal for each compound and the corresponding concentration were calculated. Excitation signal value = test group signal value - negative control signal value. Stronger excitation signal values indicate higher binding activity for forming a ternary complex.

[0452] Experimental Example 2: Compound degradation test of KRAS-G12D protein in cells

[0453] Test system and parameters:

[0454] Cells: Human colon cancer cell line GP2D

[0455] Protein quantification: Pierce TM BCA Protein Assay Kit(Thermofisher)

[0456] Protein electrophoresis: Electrophoresis tank (Thermo) 180V constant voltage electrophoresis

[0457] Protein transfer: transfer tank (BIO-RAD) 300mA constant flow transfer

[0458] Test steps

[0459] (1) Compound-treated cells

[0460] The cells were plated in six-well plates with 1.5 × 10 cells per well. 6 Cells were cultured overnight and treated with compounds (at concentrations of 0, 5, 50, and 500 nM) for 24 h.

[0461] (2) Sample processing

[0462] 1) Discard the culture medium and add 2 ml of PBS to wash the cells. Discard the wash solution and add an appropriate amount of lysis buffer (9803). Lyse the cells on ice for 30 minutes. Centrifuge at 12,000 rpm for 10 minutes at 4°C and remove the supernatant.

[0463] 2) Referring to the BCA kit instructions, prepare 40 μL of the standard protein concentration (2 mg / mL) and lysate in a 1:1 gradient dilution to 1, 0.5, 0.25, 0.125, and 0.0625 mg / mL. Add 10 μL of the sample supernatant dilution and the standard protein dilution to each well of a 96-well plate. Add 200 μL of a 50:1 mixture of reagents A and B to each well. Incubate at 37°C with shaking for 30 minutes, then measure the OD at 562 nm. Fit the standard curve, calculate the sample concentration based on the curve, and adjust the samples to the same concentration.

[0464] 3) Add 5X SDS loading buffer (Biyuntian, P0015L) to the sample and denature the protein at 100°C for 10 minutes.

[0465] (3) Protein separation, transfer, and development

[0466] 1) Protein samples were separated by electrophoresis at 180 V using NuPAGE 4-12% gradient protein precast gels (Thermofisher). The gels were transferred to membranes using a transfer tank at 4°C and 300 mA for 90 minutes and blocked with 5% skim milk powder for 1 hour.

[0467] 2) Use Ras (G12D mutation-specific) antibody (CST, 14429) and α-Tubulin antibody (CST, 2125S) diluted according to the antibody instructions and incubated with the membrane overnight at 4°C. Wash three times with TBST, 10 minutes each. Use goat anti-rabbit HRP secondary antibody (CST, 7074) diluted according to the antibody instructions and incubated with the membrane for 1 hour at room temperature. Wash three times with TBST, 10 minutes each.

[0468] 3) The membrane was immersed in ECL luminescent solution (Millipore\WBKLS0500) and the fluorescence signal was collected using a Bio-rad gel imager.

[0469] Data processing

[0470] Image J software was used to quantify the result images, and the degradation rate (KRAS-G12D) was calculated by comparing with the control group without compound addition. The calculation formula is as follows:

[0471] Degradation rate (%) = (1-(KRAS G12D (experimental group intensity × α-Tubulin control group intensity) / (α-Tubulin experimental group intensity × KRAS G12D Control group intensity)) × 100%

[0472] Test Example 3: Test of the cell proliferation inhibitory activity of the compound

[0473] Test system and procedures

[0474] AsPC-1 cells and GP2D cells were cultured in RPMI1640 with 10% fetal bovine serum (FBS) and DMEM with 10% FBS, respectively, in a 37°C CO2 incubator. Cells were plated in 96-well flat-bottomed cell culture plates, with 2,000 cells per well, and cultured overnight. The next day, compound dilutions were added. Compounds were diluted to a maximum concentration of 100 μM, and a 4-fold concentration gradient was used to dilute the compound to 9 concentration points. DMSO was used as a control. The final DMSO concentration in the cell culture medium was 1%.

[0475] The compounds were incubated with AsPC-1 cells and GP2D cells in a carbon dioxide incubator at 37°C for 5 days, and then the fluorescence signals were detected using CellCounting-LiteTM 2.0 reagent (Nanjing Novozymes Co., Ltd.).

[0476] Data processing

[0477] The inhibitory activity of the compound was calculated by the following formula: Inhibition rate = 100% - (test group / solvent group) * 100%. The curve was fitted using a four-parameter model using SigmaPlot 12.5 software to calculate the half-maximal inhibitory concentration (IC 50 ).

[0478] Test results

[0479] The effects of the compounds on KRAS were determined according to the above method. G12D The results of the proliferation inhibitory activity of mutant cells are shown in Tables 1 and 2.

[0480] Table 1. Effects of Compounds on KRAS G12D Proliferation inhibitory activity of mutant cells (AsPC-1)

[0481]

[0482]

[0483] Table 2. Effects of Compounds on KRAS G12D Proliferation inhibitory activity of mutant cells (GP2D)

[0484]

[0485] Test conclusion

[0486] The compounds of the present invention have an effect on KRAS G12D The mutant cells showed stronger proliferation inhibitory activity.

[0487] Experimental Example 4: Pharmacokinetic (PK) study in mice

[0488] Balb / c mice were administered the compound of the present application intravenously (IV) to investigate its pharmacokinetic characteristics. The IV dose was 1 mg / kg, and the IV solvent was a mixture of 5% DMSO, 5% Solutol (polyethylene glycol-15 hydroxystearate), and 90% normal saline. 40 to 50 μL of venous blood was collected before IV administration (0 h) and at 0.083 h, 0.25 h, 0.5 h, 1 h, 2 h, 4 h, 6 h, 8 h, and 24 h after administration. The blood was anticoagulated with EDTA.K2, centrifuged, and plasma samples were obtained and stored at -80 ° C. The plasma samples were treated with precipitated proteins and then subjected to LC-MS / MS analysis.

[0489] Experimental results

[0490] WinNonlin 6.3 software was used to calculate the pharmacokinetic parameters using a non-compartmental model. The results are shown in Table 3.

[0491] Table 3. Pharmacokinetic parameters of IV-administered compounds in Balb / c mice

[0492]

[0493] Test conclusion

[0494] The compounds of the present invention have excellent half-life and drug exposure in Balb / c mice when administered IV at a dose of 1 mg / kg.

[0495] The above embodiments do not limit the scope of the present invention in any way. In addition to those described herein, various modifications of the present invention will be apparent to those skilled in the art based on the foregoing description. Such modifications are also intended to fall within the scope of the appended claims. Each reference cited in this application (including all patents, patent applications, journal articles, books and any other disclosures) is incorporated herein by reference in its entirety.

Claims

1. A compound or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite or prodrug thereof, wherein the compound has the structure of formula (I): in, Ring A is a 3-7 membered heterocyclic group, which is optionally substituted by one or more R 7 replace; Ring E is a 6-12 membered spiro heterocyclic group, a 6-12 membered spiro heterocyclic group or a 6-12 membered bridged heterocyclic group, which is optionally substituted by one or more R 8 replace; Ring X is phenyl or 5-6 membered heteroaryl, which is optionally substituted by one or more R 9 replace; Ring Y is a 5-6 membered heteroaryl group; Ring Z is phenyl or 5-6 membered heteroaryl, which is optionally substituted by one or more R 10 replace; L 1 is a covalent bond, or is selected from O, S and NR 11 ; L 2 is a covalent bond, or is selected from O, S and NR 12 ; L 3 Selected from C 1-6 Alkylene, -OC 1-6 Alkylene-, -C 1-6 Alkylene-O- and -C 1-6 Alkylene-OC 1-6 Alkylene-; R 1 Selected from hydrogen, halogen, cyano, C 1-6 Alkyl, C 2-6 Alkenyl, C 3-6 Cycloalkyl, C 1-6 Haloalkyl, -OC 1-6 Alkyl, -OC 1-6 Haloalkyl and -OC 3-6 Cycloalkyl; R 2 Selected from hydrogen, halogen, cyano, C 1-6 Alkyl, C 2-6 Alkenyl, C 3-6 Cycloalkyl, C 1-6 Haloalkyl, -OC 1-6 Alkyl, -OC 1-6 Haloalkyl and pentafluorosulfur; R 3 are independently selected from hydrogen, halogen, cyano, C 1-6 Alkyl, C 1-6 Haloalkyl and C 3-6 Cycloalkyl; R 4 Selected from C 1-6 Alkyl, C 1-6 Haloalkyl and C 3-6 Cycloalkyl; R 5a and R 5b are each independently selected from hydrogen, C 1-6 Alkyl, C 1-6 Haloalkyl, -C 1-6 Alkylene -OH and C 3-6 Cycloalkyl; or R 5a and R 5b Together with the carbon atoms connected to form C 3-6 Cycloalkyl or 3-6 membered heterocyclic group; R 6 are independently selected from hydrogen, halogen, cyano, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 3-6 Cycloalkyl, -OC 1-6 Alkyl and -OC 1-6 alkyl halide; R 7 Selected from hydrogen, C 1-6 Alkyl, C 1-6 Haloalkyl and C 3-6 Cycloalkyl; R 8 Selected from hydrogen, halogen, C 1-6 Alkyl, C 1-6 Haloalkyl and C 3-6 Cycloalkyl; R 9 Selected from hydrogen, halogen, C 1-6 Alkyl, C 1-6 Haloalkyl and C 3-6 Cycloalkyl; R 10 Selected from hydrogen, halogen, cyano, C 1-6 Alkyl and C 1-6 alkyl halide; R 11 Selected from hydrogen, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 3-6 Cycloalkyl, 3-6 membered heterocyclic group, -C 1-6 Alkylene-C 3-6 Cycloalkyl and -C 1-6 Alkylene-3-6 membered heterocyclic group; R 12 Selected from hydrogen and C 1-6 alkyl; m is selected from 0, 1, 2, 3 and 4; n is selected from 0, 1, 2, 3 and 4.

2. The compound of claim 1 or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotope-labeled compound, metabolite or prodrug thereof, wherein: L 2 is a covalent bond, or is selected from O, S and NH; Ring E is a 6-12 membered spiro heterocyclic group, a 6-12 membered spiro heterocyclic group or a 6-12 membered bridged heterocyclic group, optionally substituted by one or more R 8 replace; R 8 is selected from hydrogen, fluorine, chlorine, bromine, methyl, ethyl, isopropyl, trifluoromethyl and cyclopropyl; Preferably, L 2 is a covalent bond, or is selected from O, S and NH; Ring E is selected from 6-12 membered nitrogen-containing spiro heterocyclic group, 6-12 membered oxygen-containing spiro heterocyclic group, which is optionally substituted by one or more R 8 Replacement; R 8 is selected from hydrogen, fluorine, chlorine, bromine, methyl, ethyl, isopropyl, trifluoromethyl and cyclopropyl; Preferably, L 2 is a covalent bond, or is selected from O, S and NH; Ring E is selected from 6-10 membered nitrogen-containing spiro heterocyclic groups and 6-10 membered oxygen-containing spiro heterocyclic groups, which are optionally substituted by one or more R 8 Replacement; R 8 is selected from hydrogen, fluorine, chlorine, bromine, methyl, ethyl, isopropyl, trifluoromethyl and cyclopropyl; Preferably, L 2 is a covalent bond or O; Ring E is selected from More preferably, the fragment Selected from 3. The compound of claim 1 or 2, or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotope-labeled compound, metabolite or prodrug thereof, wherein: L 1 is a covalent bond, or is selected from O and NR 11 ; Ring A is a 5-membered heterocyclic group, which is optionally substituted by one or more R 7 replace; R 7 is selected from hydrogen, methyl, ethyl, isopropyl, trifluoromethyl and cyclopropyl; R 11 Selected from hydrogen, C 1-3 Alkyl, C 1-3 Halogenated alkyl, C 3-6 Cycloalkyl, -C 1-3 Alkylene-C 3-6 Cycloalkyl and -C 1-3 Alkylene-3-6 membered heterocyclic group; Preferably, L 1 Selected from O and NR 11 ; Ring A is a 5-membered nitrogen-containing heterocyclic group; R 11 Selected from hydrogen, C 1-3 Alkyl, C 1-3 Haloalkyl and C 3-6 Cycloalkyl; Preferably, L 1 Selected from O and NR 11 ; Ring A is pyrrolyl; R 11 is selected from hydrogen, methyl, ethyl, isopropyl, trifluoromethyl and cyclopropyl; More preferably, the fragment Selected from 4. The compound of any one of claims 1 to 3, or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotope-labeled compound, metabolite or prodrug thereof, wherein: n is 1; R 6 selected from halogen; Preferably, n is 1; R 6 is selected from the group consisting of hydrogen, fluorine, chlorine and bromine; Preferably, the fragment for 5. The compound according to any one of claims 1 to 4, or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotope-labeled compound, metabolite or prodrug thereof, wherein R 1 is selected from hydrogen, methyl, fluorine, chlorine and methoxy; preferably, R 1 For hydrogen.

6. The compound of any one of claims 1 to 5, or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotope-labeled compound, metabolite or prodrug thereof, wherein R 2 is selected from hydrogen, fluorine, chlorine, bromine, methyl, ethyl, vinyl, trifluoromethyl and cyclopropyl; preferably, R 2 It is cyclopropyl.

7. The compound of any one of claims 1 to 6, or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotope-labeled compound, metabolite or prodrug thereof, wherein: m is selected from 0, 1 and 2; R 3 each independently selected from hydrogen, fluorine, chlorine, bromine, methyl, ethyl, isopropyl, trifluoromethyl and cyclopropyl; Preferably, m is 2; R 3 are each independently selected from fluoro and methyl; More preferably, snippet for 8. The compound of any one of claims 1 to 7, or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotope-labeled compound, metabolite or prodrug thereof, wherein L 3 Selected from C 1-3 Alkylene, -OC 1-3 Alkylene-, -C 1-3 Alkylene-O- and -C 1-3 Alkylene-OC 1-3 Alkylene-; preferably, L 3 is selected from -CH2-CH2-, -O-CH2-, -CH2-O- and -CH2-O-CH2-; more preferably, L 3 It is -O-CH2-.

9. The compound according to any one of claims 1 to 8, or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotope-labeled compound, metabolite or prodrug thereof, wherein ring X is phenyl or pyridyl; preferably, ring X is phenyl; more preferably, ring X is 10. The compound of any one of claims 1 to 9, or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotope-labeled compound, metabolite or prodrug thereof, wherein ring Y is a 5-membered heteroaromatic ring; preferably, ring Y is a 5-membered nitrogen-containing heteroaromatic ring; preferably, ring Y is pyrazolyl, triazolyl or isoxazolyl; preferably, ring Y is triazolyl; more preferably, ring Y is 11. The compound of any one of claims 1 to 10, or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotope-labeled compound, metabolite or prodrug thereof, wherein R 4 Selected from C 1-6 Alkyl and C 3-6 Cycloalkyl; preferably, R 4 Selected from C 3-6 Alkyl and C 3-6 Cycloalkyl; preferably, R 4 is selected from isopropyl, tert-butyl and cyclohexyl; preferably, R 4 is selected from isopropyl and tert-butyl.

12. The compound of any one of claims 1 to 11, or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotope-labeled compound, metabolite or prodrug thereof, wherein R 5a and R 5b are each independently selected from hydrogen, methyl, ethyl, isopropyl, -CH2-OH and cyclopropyl; preferably, R 5a and R 5b are each independently selected from hydrogen and -CH2-OH; more preferably, R 5a is hydrogen, R 5b It is -CH2-OH.

13. The compound of any one of claims 1 to 12, or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite, or prodrug thereof, wherein: Ring Z is phenyl or pyridyl, which is optionally substituted by one or more R 10 replace; R 10 is selected from the group consisting of hydrogen, fluorine, chlorine, bromine, methyl, ethyl and trifluoromethyl; Preferably, ring Z is phenyl; More preferably, ring Z is 14. A compound according to any one of claims 1 to 13, or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite, or prodrug thereof, wherein the compound has a structure of Formula (II-a) or Formula (II-b): Among them, ring E, L 2 、R 4 and R 11 As defined in any one of claims 1 to 13; Preferably, the compound has the structure of formula (III-a), formula (III-b), formula (III-c) or formula (III-d): Among them, ring E, L 2 、R 4 and R 11 As defined in any one of claims 1 to 13.

15. The compound of claims 1-14, or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite, or prodrug thereof, wherein the compound is selected from: Preferably, the compound is selected from:

16. A pharmaceutical composition comprising a prophylactically and / or therapeutically effective amount of a compound according to any one of claims 1 to 15, or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotope-labeled compound, metabolite or prodrug thereof, and one or more pharmaceutically acceptable carriers.

17. Use of the compound according to any one of claims 1 to 15 or its pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotope-labeled compound, metabolite or prodrug, or the composition according to claim 16 in the preparation of a medicament for preventing and / or treating KRAS G12D-mediated related diseases.

18. A method for preparing the compound of claim 1, wherein L 3 is -O-CH2-, and ring Y is When it is as shown in Route 1: Route 1 Among them, ring A, ring E, ring X, ring Z, R 1 、R 2 、R 3 、R 4 、R 5a 、R 5b 、R 6 , L 1 , L 2 , m and n are as defined in claim 1; LG 1 LG 2 LG 3 LG 4 LG 5 LG 6 LG 7 and LG 8 each independently represents a leaving group, such as a halogen, a methylsulfonate, a thiomethyl, a methylsulfoxide, a methylsulfone, a boronic acid, a borate ester, a methoxy group, or an ethoxy group; PG 1 and PG 2 represents a protecting group, for example, benzyloxycarbonyl (Cbz), tert-butyloxycarbonyl (Boc), tetrahydropyranyl (THP), tert-butyl, (S)-1-phenylethanol or benzyl; The method comprises the following steps: 1) reacting compound I-1 with compound I-2 to obtain compound I-3; 2) reacting compound I-3 with compound I-4 to obtain compound I-5; 3) reacting compound I-5 with compound I-6 to obtain compound I-7; 4) subjecting compound I-7 to a coupling reaction with compound I-8 to obtain compound I-9; 5) subjecting compound I-9 to a coupling reaction with compound I-10 to obtain compound I-11; 6) subjecting compound I-11 to a deprotection reaction to obtain compound I-12; 7) reacting compound I-12 with compound I-13 to obtain compound I-14; 8) reacting compound 1-14 with compound 1-15 to obtain compound 1-16; and 9) subjecting compound I-16 to a deprotection reaction to obtain compound I; Or, when L 3 is -O-CH2-, and ring Y is When, the method is as shown in Route 2: Route 2 Among them, ring A, ring E, ring X, ring Z, R 1 、R 2 、R 3 、R 4 、R 5a 、R 5b 、R 6 , L 1 , L 2 , m and n are as defined in claim 1; LG 1 LG 2 LG 3 LG 4 LG 5 LG 6 LG 7 and LG 8 each independently represents a leaving group, such as a halogen, a methylsulfonate, a thiomethyl, a methylsulfoxide, a methylsulfone, a boronic acid, a borate ester, a methoxy group, or an ethoxy group; PG 1 and PG 2 represents a protecting group, for example, benzyloxycarbonyl (Cbz), tert-butyloxycarbonyl (Boc), tetrahydropyranyl (THP), tert-butyl, (S)-1-phenylethanol or benzyl; The method comprises the following steps: 1) reacting compound I-1 to obtain compound I-2a; 2) reacting compound I-2a to obtain compound I-3a; 3) reacting compound I-3a to obtain compound I-4a; 4) reacting compound I-4a with compound I-6 to obtain compound I-5a; 5) coupling reaction of compound I-5a with compound I-8 to obtain compound I-6a; 6) coupling reaction of compound I-6a with compound I-10 to obtain compound I-7a; 7) Compound I-7a is reacted to obtain compound I-8a 8) subjecting compound I-8a to a deprotection reaction to obtain compound I-9a; 9) reacting compound I-9a with compound I-13 to obtain compound I-10a; 10) reacting compound I-10a to obtain compound I-11a; 11) reacting compound I-11a to obtain compound I-12a; 12) reacting compound I-12a to obtain compound I-14; 13) reacting compound 1-14 with compound 1-15 to obtain compound 1-16; and 14) Compound I-16 is subjected to a deprotection reaction to obtain Compound I.

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