WO2023280026A1 - Heteroaromatic ring compound, preparation method therefor and use thereof - Google Patents

Abstract

A heteroaromatic ring compound, a preparation method therefor and the use thereof. Specifically, the present invention relates to a compound of formula (I) or a pharmaceutically acceptable salt, a stereoisomer, a tautomer, a polymorph, a solvate, an N-oxide, an isotope labeled compound, a metabolite or a prodrug thereof, a pharmaceutical composition containing same, a preparation method therefor, and the use thereof in the preparation of a drug for preventing or treating related diseases mediated by KRAS G12D.

Description

Heteroaromatic compound, its preparation method and use technical field

The invention belongs to the field of medicine, and specifically relates to a class of heteroaromatic compounds, a preparation method and application thereof.

Background technique

RAS is a guanine nucleotide binding protein with GTPase activity and has the function of anchoring inside the cell membrane. RAS protein can switch between the GDP-bound inactive state and the GTP-bound active state, affecting multiple signaling pathways such as downstream Raf, PI3K, and RalGDS, and regulating protein synthesis, gene transcription, cell growth, differentiation, apoptosis, and migration, etc. .

RAS mutations will lead to the continuous activation of downstream signaling pathways and promote the occurrence and development of tumors. In all tumor types, RAS mutations mainly occur in KRAS (85%), and the 12-position glycine on KRAS is mutated to aspartic acid (G12D ) has the highest frequency (34.2%). In tumor cells with KRAS mutation, the GTPase activity of KRAS decreases and remains active. KRAS mutations are closely related to the occurrence of lung cancer, pancreatic cancer and colorectal cancer.

The development of small molecule inhibitors of KRAS G12D is in its infancy. WO2020173935 discloses a class of RAS inhibitors, which use the principle of molecular glue to induce KRAS G12D to form dimers and block the interaction between KRAS and downstream proteins. In addition, WO2021041671 discloses a new class of KRAS G12D inhibitors, but does not disclose its specific mechanism of action.

Proteolysis-Targeting Chimeras (PROTACs) are bifunctional molecules whose structure includes three parts: (1) the part that binds to the target protein substrate; (2) the part that binds to the E3 ubiquitin ligase and (3) the 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 E3 ubiquitin ligase, induce the recruitment of E3 ubiquitin ligase to the surface of the target protein, trigger the ubiquitination process and induce the degradation of the target protein . PROTAC molecules have a wide range of pharmacological activities, high target selectivity, can be used to degrade difficult-to-drug target proteins, have strong degradation efficiency, and can maintain catalytic degradation at low concentrations.

There are currently no PROTAC molecules that specifically target KRAS G12D. In this field, there is an urgent need to develop KRAS G12D PROTAC molecules with novel structure and good biological activity.

Summary of the invention

The present invention provides compounds useful as degraders and/or inhibitors targeting KRAS G12D protein. The compound of the present invention belongs to PROTAC, one end of which can bind to target protein, and the high-affinity VHL ligand at the other end can recruit target protein to E3 ubiquitin ligase, resulting in ubiquitination and subsequent degradation of KRAS G12D protein. The compounds of the present invention can be used to treat and/or prevent cancers or disorders caused by aggregation and/or overactivation of target proteins.

In one aspect, the invention provides a compound, or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite or precursor thereof Drug, said compound has the structure of formula (I):

in:

为单键或双键；

is a single or double bond;

为单键时，X ¹选自C(R ¹) ₂和C(O)；且X ²选自N和CR ¹； When between X ¹ and X ²

When it is a single bond, X ¹ is selected from C(R ¹ ) ₂ and C(O); and X ² is selected from N and CR ¹ ;

为双键时，X ¹选自CR ¹和N；且X ²为C； When between X ¹ and X ²

When it is a double bond, X ¹ is selected from CR ¹ and N; and X ² is C;

为单键时，X ³和X ⁴各自独立地选自C(R ¹) ₂、NR ⁸和C(O)； When between X ³ and X ⁴

When it is a single bond, X ³ and X ⁴ are each independently selected from C(R ¹ ) ₂ , NR ⁸ and C(O);

为双键时，X ³和X ⁴各自独立地选自CR ¹和N； When between X ³ and X ⁴

When it is a double bond, X ³ and X ⁴ are each independently selected from CR ¹ and N;

Each R ¹ is independently selected from hydrogen, halogen, C _1-6 alkyl, C _1-6 alkenyl, C _3-6 cycloalkyl, C _1-6 haloalkyl, -OC _1-6 alkyl, - OC _1-6 haloalkyl and -OC _3-6 cycloalkyl; or two R ¹ on the same carbon atom form carbonyl, C _3-6 cycloalkyl or 3-6 membered heterocycle with the carbon atom they are connected to base;

R ⁸ is independently selected from hydrogen, C _1-3 alkyl, C _3-6 cycloalkyl and C _1-3 haloalkyl;

R ² is selected from C _6-10 aryl or 5-10 membered heteroaryl, each of said aryl and heteroaryl is optionally substituted by one or more R ³ ;

When present, each R ³ is independently selected from halogen, hydroxy, cyano, C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl, C _1-6 haloalkyl, -C _{1 -6} alkyl-OH, -OC _1-6 alkyl, C _3-6 cycloalkyl, 3-10 membered heterocyclyl and 5-10 membered heteroaryl, the alkyl, alkenyl, alkynyl, Cycloalkyl, heterocyclyl and heteroaryl are each optionally substituted by one or more substituents selected from halogen, hydroxy, cyano, C _1-6 alkyl and C _1-6 haloalkyl;

L is -L ¹ -L ² -L ³ -L ⁴ -L ⁵ -L ⁶ -;

L ¹ is selected from -O-, -S-, -NR ^1a -, C _1-6 alkylene, C _3-6 cycloalkylene and 3-10 membered heterocyclylene, said alkylene, alkylene Cycloalkyl and heterocyclylene are each optionally substituted by one or more substituents selected from halogen, hydroxy, cyano, C _1-6 alkyl and C _1-6 haloalkyl;

L ² is a covalent bond, or selected from -O-, -NR ^2a -, C _1-6 alkylene, -OC _1-6 alkylene, C _1-6 alkylene-O-, C _{3- 6} cycloalkylene groups and 3-10 membered heterocyclylene groups, each of the alkylene, cycloalkylene and heterocyclylene groups is optionally selected from one or more halogen, hydroxyl, cyano, C ₁ Substituents of _-6 alkyl and C _1-6 haloalkyl;

L ³ is a covalent bond, or selected from -O-, -NR ^3a -, C _1-6 alkylene, -OC _1-6 alkylene, C _1-6 alkylene-O-, C _{3- 6} cycloalkylene groups and 3-10 membered heterocyclylene groups, each of the alkylene, cycloalkylene and heterocyclylene groups is optionally selected from one or more halogen, hydroxyl, cyano, C ₁ Substituents of _-6 alkyl and C _1-6 haloalkyl;

L ⁴ is a covalent bond, or is selected from -O-, -NR ^4a -, C _1-6 alkylene, -OC _1-6 alkylene, C _1-6 alkylene-O-, C _{3- 6} cycloalkylene groups and 3-10 membered heterocyclylene groups, each of the alkylene, cycloalkylene and heterocyclylene groups is optionally selected from one or more halogen, hydroxyl, cyano, C ₁ Substituents of _-6 alkyl and C _1-6 haloalkyl;

L ⁵ is a covalent bond, or selected from -O-, -NR ^5a -, C _1-6 alkylene, -OC _1-6 alkylene-, C _1-6 alkylene-O-, C _{3 -6} cycloalkylene and 3-10 membered heterocyclylene, said alkylene, cycloalkylene and heterocyclylene are each optionally replaced by one or more selected from halogen, hydroxyl, cyano, C Substituents of _1-6 alkyl and C _1-6 haloalkyl;

L ⁶ is selected from -O-, -S-, -NR ^6a -, C _1-6 alkylene, -OC _1-6 alkylene-, C _1-6 alkylene-O-, C _3-6 Cycloalkylene, -C(O)-, -C(O)-N(R ^6a )- and 3-10 membered heterocyclylene, each of the alkylene, cycloalkylene and heterocyclylene optionally substituted by one or more substituents selected from halogen, hydroxyl, cyano, C _1-6 alkyl and C _1-6 haloalkyl;

R ^1a , R ^2a , R ^3a , R ^4a , R ^5a and R ^6a are each independently selected from hydrogen and C _1-6 alkyl;

M is a small molecule ligand of E3 ubiquitin ligase VHL, preferably its structure is shown in the following formula:

in

A is selected from -NH-, -EC(=O)-NR ^7a -, -E-NR ^7a -C(=O)-, -EC(=O)- and 5-10 membered heteroarylene, said Heteroarylene is optionally substituted by one or more substituents selected from halogen, hydroxy, cyano, C _1-6 alkyl and C _1-6 haloalkyl;

E is selected from C _1-6 alkylene, C _3-6 cycloalkylene and 3-10 membered heterocyclylene, each of which is optionally replaced by one Or more substituents selected from halogen, hydroxyl, cyano, C _1-6 alkyl and C _1-6 haloalkyl;

R ^7a is selected from hydrogen and C _1-6 alkyl;

R ⁴ is selected from hydrogen, C _1-6 alkyl, C _3-6 cycloalkyl and 3-12 membered heterocyclyl;

R ^is selected from hydrogen, C _1-6 alkyl, C _3-6 cycloalkyl, 3-12 membered heterocyclyl, each of which is optionally replaced by one or more Substituents selected from halogen, hydroxyl, cyano, C _1-6 alkyl and C _1-6 haloalkyl;

R is selected from hydrogen, halogen, hydroxyl, C ^1-6 _alkyl , C _2-6 alkenyl and C _2-6 alkynyl; and

表示连接位点。

Indicates the junction site.

In another aspect, the present invention provides a pharmaceutical composition comprising a prophylactically and/or therapeutically effective amount of the compound of the present invention or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, Solvates, N-oxides, isotope-labeled compounds, metabolites or prodrugs, and one or more pharmaceutically acceptable carriers.

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 compounds, metabolites or prodrugs, or pharmaceutical compositions comprising the invention.

In another aspect, the present invention provides a compound of the present invention, or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, or a pharmaceutically acceptable salt thereof, Metabolites or prodrugs, or pharmaceutical compositions of the present invention, which target the KRAS G12D protein and are used to degrade the KRAS G12D protein.

In another aspect, the present invention provides a compound of the present invention, or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, or a pharmaceutically acceptable salt thereof, Metabolites or prodrugs, or pharmaceutical compositions of the present invention, which are used to prevent and/or treat KRAS G12D-mediated related diseases.

In another aspect, the present invention provides a compound of the present invention, or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, or a pharmaceutically acceptable salt thereof, Use of metabolites or prodrugs or the pharmaceutical composition of the present invention in the preparation of medicines for preventing and/or treating KRAS G12D-mediated related diseases.

In another aspect, the present invention provides a method for preventing and/or treating related diseases mediated by KRAS G12D, which comprises administering a preventive and/or therapeutically effective amount of a compound of the present invention or a pharmaceutically acceptable amount thereof to an individual in need thereof. Salts, stereoisomers, tautomers, polymorphs, solvates, N-oxides, isotope-labeled compounds, metabolites or prodrugs, or pharmaceutical compositions of the present invention.

In another aspect, the invention provides methods for preparing the compounds of the invention.

Detailed description of the invention

definition

Unless defined otherwise hereinafter, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. References to techniques used herein are intended to refer to techniques commonly understood in the art, including those variations of the techniques or substitutions of equivalent techniques that are obvious to those skilled in the art. While the following terms are believed to be well understood by those skilled in the art, the following definitions are set forth to better explain the present invention.

As used herein, the terms "comprising", "comprising", "having", "containing" or "involving" and other variations thereof herein are inclusive or open-ended, and do not Other unlisted elements or method steps are excluded.

As used herein, the term "alkyl" is defined as a straight or branched chain 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), optionally substituted by one or more (such as 1, 2 or 3) suitable substituents such as halogen.

As used herein, the term "alkylene" refers to a divalent saturated aliphatic hydrocarbon group obtained by removing one more hydrogen atom from the corresponding "alkyl". For example, as used herein, the term "C _1-6 alkylene" refers to an alkylene group having 1 to 6 carbon atoms, such as methylene (-CH ₂ -), ethylene (-CH ₂ CH ₂ -), isopropylidene (-CH( _CH3 )CH2- ₎ , optionally substituted by one or more (such as 1, 2 or 3) suitable substituents such as halogen.

As used herein, the term "alkenyl" refers to a straight or branched chain aliphatic hydrocarbon group having one or more carbon-carbon double bonds. For example, the term "C alkenyl" as used herein 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-hexene group, 4-hexenyl, 5-hexenyl, 2-methyl-2-propenyl or 4-methyl-3-pentenyl, etc.), which is optionally replaced by one or more (such as 1, 2 or 3) suitable substituents such as halogen substitution.

As used herein, the term "alkynyl" refers to a straight or branched chain aliphatic hydrocarbon group having one or more carbon-carbon triple bonds. For example, the term " _C2-6 alkynyl" as used herein 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.

As used herein, the term "cycloalkyl" refers to a saturated or partially unsaturated non-aromatic monocyclic or polycyclic (such as bicyclic) hydrocarbon ring (e.g. monocyclic, such as cyclopropyl, cyclobutyl, cyclopentyl , cyclohexyl, cycloheptyl, cyclooctyl or cyclononyl, or bicyclic rings, including spiro, 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, decalinyl, 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 (such as cyclopropyl, cyclo butyl, cyclopentyl or cyclohexyl) optionally substituted by one or more (such as 1, 2 or 3) suitable substituents, eg methyl substituted cyclopropyl.

As used herein, the term "cycloalkylene" refers to a divalent group obtained by removing one more hydrogen atom from the corresponding "cycloalkyl".

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

As used herein, the term "haloalkyl" refers to an alkyl group substituted with one or more (such as 1, 2 or 3) same or different halogen atoms. For example, the term "C _1-6 haloalkyl" refers to a haloalkyl group having 1 to 6 carbon atoms, such as -CF ₃ , -C ₂ F ₅ , -CHF ₂ , -CH ₂ F, -CH ₂ CF ₃ , - _{CH2Cl or -CH2CH2CF3 .}

As used herein, the term "heterocyclyl" refers to a saturated or partially unsaturated monocyclic or polycyclic group, for example having 2, 3, 4, 5, 6, 7, 8 or 9 carbons in the ring atom and one or more (for example 1, 2, 3 or 4) heteroatoms independently selected from N, O or S(O) _t (wherein t is 0, 1 or 2), for example 3- 12-membered heterocyclic group, 3-7-membered heterocyclic group, 3-6-membered heterocyclic group, 5-6-membered heterocyclic group, etc. Representative examples of heterocyclyl include, but are not limited to, oxiranyl, aziridinyl, azetidinyl, oxetanyl, tetrahydrofuryl, tetrahydropyrrolidinyl, hexa Hydrogen-1H-pyrroline, pyrrolidonyl, imidazolidinyl, pyrazolidinyl, tetrahydropyranyl, piperidinyl, morpholinyl, dithianyl, thiomorpholinyl or piperazinyl.

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

As used herein, the term "aryl" or "aromatic ring" refers to an all-carbon monocyclic or fused-ring polycyclic aromatic group having a conjugated pi-electron system. For example, the term "C _6-10 aryl" or "C _6-10 aryl ring" refers to an aromatic group containing 6 to 10 carbon atoms, such as phenyl (ring) or naphthyl (ring). Aryl is optionally substituted with 1 or more (such as 1, 2 or 3) suitable substituents (eg halogen, -OH, -CN, -NO ₂ or C _1-6 alkyl, etc.).

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, especially containing 1, 2, 3, 4, 5, 6, 9 or 10 carbon atoms, and additionally in each case may be Benzo-fused. For example, a heteroaryl or heteroaryl ring may be selected from thienyl (ring), furyl (ring), pyrrolyl (ring), oxazolyl (ring), thiazolyl (ring), imidazolyl (ring), pyryl Azolyl (ring), isoxazolyl (ring), isothiazolyl (ring), oxadiazolyl (ring), triazolyl (ring), thiadiazolyl (ring), and their benzos Derivatives; or pyridyl (ring), pyridazinyl (ring), pyrimidinyl (ring), pyrazinyl (ring), triazinyl (ring), etc., and their benzo derivatives.

As used herein, the term "heteroarylene" refers to a divalent group obtained by removing one more hydrogen atom from the corresponding "heteroaryl".

The term "substituted" means that one or more (eg, 1, 2, 3 or 4) hydrogens on the indicated atom are replaced by a selection from the indicated group, provided that no more than the indicated atom is in the current instance normal valences and the substitutions result in stable compounds. Combinations of substituents and/or variables are permissible only if such combinations result in stable compounds.

If a substituent is described as being "optionally substituted by", the substituent can be (1) unsubstituted or (2) substituted. If a carbon of a substituent is described as being optionally substituted with one or more of the list of substituents, one or more hydrogens on the carbon (to the extent of any hydrogen present) may be independently and/or together Substituents of choice are either substituted or unsubstituted. If the nitrogen of a substituent is described as being optionally substituted with one or more of the list of substituents, one or more hydrogens on the nitrogen (to the extent of any hydrogen present) may each be independently selected substituents Substituted or not.

If substituents are described as being "independently selected from" a group, each substituent is selected independently of the other. Accordingly, each substituent may be the same as or different from another (other) substituent.

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.

As used herein, unless otherwise indicated, the point of attachment of a substituent may be from any suitable position of the substituent.

When a bond for a substituent is shown as being through a bond connecting two atoms in a ring, then such substituent may be bonded to any ring-forming atom in such a substitutable ring.

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 have been labeled with the same atomic number but an atomic mass or mass number different from the atomic mass prevailing in nature. or mass number of atomic substitutions. Examples of isotopes suitable for inclusion in compounds of the invention include, but are not limited to, isotopes of hydrogen (e.g. ² H, ³ H, deuterium D, tritium T); isotopes of carbon (e.g. ¹¹ C, ¹³ C, and ¹⁴ C); chlorine isotopes of fluorine (such as ¹⁸ F); ^isotopes of iodine (such as ¹²³ I and ¹²⁵ I); isotopes of nitrogen (such as ¹³ N and ¹⁵ N); isotopes of oxygen (such as ¹⁵ O, ¹⁷ O and ¹⁸ O); isotopes of phosphorus (eg ³² P); and isotopes of sulfur (eg ³⁵ S). Certain isotopically-labeled compounds of the invention (eg, those incorporating radioactive isotopes) are useful in drug and/or substrate tissue distribution studies (eg, assays). The radioisotopes tritium ( ^ie3H ) and carbon- ¹⁴ (ie14C) are particularly useful for this purpose because of their ease of incorporation and ease of detection. Substitution with positron-emitting isotopes such as ¹¹ C, ¹⁸ F, ¹⁵ O, and ¹³ N can be used in positron emission tomography (PET) studies to examine substrate receptor occupancy. Isotopically labeled compounds of the invention can be prepared by methods analogous to those described in the accompanying Schemes and/or Examples and Preparations by using an appropriate isotopically labeled reagent in place of the non-labeled reagent previously employed. Pharmaceutically acceptable solvates of the invention include those wherein the solvent of crystallization may be isotopically substituted, eg, _D2O , acetone-d6 or _{DMSO -} d6.

The term "stereoisomer" means isomers formed as a result of at least one asymmetric center. In compounds with one or more (e.g. 1, 2, 3 or 4) asymmetric centers, which can give rise to racemic mixtures, single enantiomers, diastereomeric mixtures and individual non- Enantiomers. Certain individual molecules may also exist as geometric isomers (cis/trans). Similarly, compounds of the present invention may exist as mixtures of two or more structurally distinct forms (commonly referred to as tautomers) in rapid equilibrium. Representative examples of tautomers include keto-enol tautomers, phenol-keto tautomers, nitroso-oxime tautomers, imine-enamine tautomers Wait. It is to be understood that the scope of this application encompasses all such ratios in any proportion (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%) %) isomers or mixtures thereof.

The present invention covers all possible crystalline forms or polymorphs of the compounds of the present invention, which may be a single polymorph or a mixture of more than one polymorph in any proportion.

It will also be understood that certain compounds of the present invention may exist in free form for use in therapy, or, where appropriate, as pharmaceutically acceptable derivatives thereof. In the present invention, pharmaceutically acceptable derivatives include but are not limited to: pharmaceutically acceptable salts, solvates, metabolites or prodrugs, which can directly or indirectly Compounds of the invention or metabolites or residues thereof are provided. Therefore, when a "compound of the present invention" is referred to herein, it is also intended to cover the above-mentioned various derivative forms of the compound.

The 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 which form pharmaceutically acceptable salts. Suitable base addition salts are formed from bases which form pharmaceutically acceptable salts. For a review of suitable salts see "Handbook of Pharmaceutical Salts: Properties, Selection, and Use" by Stahl and Wermuth (Wiley-VCH, 2002). Methods for preparing pharmaceutically acceptable salts of the compounds of the invention are known to those skilled in the art.

The compounds of the invention may exist in the form of solvates, preferably hydrates, wherein the compounds of the invention comprise a polar solvent, such as water, methanol or ethanol in particular, as a structural element of the crystal lattice of the compound. The amount of polar solvent, especially water, may be present in stoichiometric or non-stoichiometric ratios.

Those skilled in the art will appreciate that not all nitrogen-containing heterocycles are capable of forming N-oxides since nitrogen requires available lone pairs of electrons to oxidize to oxides; nitrogen-containing heterocycle. Those skilled in the art will also recognize that tertiary amines are capable of forming N-oxides. Synthetic methods for the preparation of N-oxides of heterocycles and tertiary amines are well known to those skilled in the art and include the use of peroxyacids such as peracetic acid and m-chloroperbenzoic acid (MCPBA), hydrogen peroxide, alkyl Hydrogen peroxides such as t-butyl hydroperoxide, sodium perborate and dioxiranes such as dimethyldioxirane are used to oxidize heterocycles and tertiary amines. These methods for preparing N-oxides have been extensively described and reviewed in the literature, see for example: T.L. Gilchrist, Comprehensive Organic Synthesis, vol.7, pp 748-750; A.R. Katritzky and A.J. Boulton, Eds., Academic Press and G.W.H. Cheeseman and E.S.G. Werstiuk, Advances in Heterocyclic Chemistry, vol.22, pp 390-392, A.R. Katritzky and A.J. Boulton, Eds., Academic Press.

Also included within the scope of the present invention are metabolites of the compounds of the present invention, ie substances formed in vivo upon administration of the compounds of the present invention. Such products may result, for example, from oxidation, reduction, hydrolysis, amidation, deamidation, esterification, enzymatic hydrolysis, etc., of the administered compound. Accordingly, the present invention includes metabolites of compounds of the present invention, including compounds produced by contacting a compound of the present invention with a mammal for a time sufficient to produce a metabolite thereof.

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 which themselves may have little or no pharmacological activity, when administered to the body or The above can be converted into compounds of the invention having the desired activity by, for example, hydrolytic cleavage. Typically such prodrugs will be functional group derivatives of the compound which are readily converted in vivo into the desired therapeutically active compound. 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 ( Edited by E.B. Roche, American Pharmaceutical Association). The prodrugs of the present invention can be obtained, for example, by using certain moieties known to those skilled in the art as "pro-moiety (such as described in "Design of Prodrugs", H. Bundgaard (Elsevier, 1985))". Prepared by substituting appropriate functional groups present in the compounds of the invention.

The invention also encompasses compounds of the invention which contain protecting groups. During any of the preparations of the compounds of the invention, it may be necessary and/or desirable to protect sensitive or reactive groups on any of the molecules involved, thereby forming chemically protected forms of the compounds of the invention. This can be achieved by conventional protecting groups, for example, as described in Protective Groups in Organic Chemistry, ed. J.F.W. McOmie, Plenum Press, 1973; and T.W. Greene & P.G.M. Wuts, Protective Groups in Organic Synthesis, John Wiley & Sons, 1991 For those protecting groups, these references are incorporated herein by reference. Protecting groups can be removed at an appropriate subsequent stage using methods known in the art.

The term "about" means within ±10%, preferably within ±5%, more preferably within ±2% of the stated value.

compound

The present invention provides compounds of formula (I) or pharmaceutically acceptable salts, stereoisomers, tautomers, polymorphs, solvates, N-oxides, isotope-labeled compounds, and metabolites thereof or prodrugs:

X ¹、X ²、X ³、X ⁴、R ²、L和M如上文所定义。 in

X ¹ , X ² , X ³ , X ⁴ , R ² , L and M are as defined above.

In some embodiments of the present invention, the compound of formula (I) above has a structure as shown in formula (IA) or (IB):

wherein R ¹ , R ² , L and M are as defined above, and n is 0, 1, 2, 3 or 4 for R ¹ other than hydrogen.

In some embodiments of the present invention, the above-mentioned compound of formula (I) has a structure as shown in formula (IC):

wherein R ¹ , R ² , L and M are as defined above, and n is 0, 1, 2, 3 or 4 for R ¹ other than hydrogen.

In some embodiments of compounds of Formula (I), Formula (IA), (IB) and (IC), each R ¹ is independently selected from hydrogen and halogen. Preferably, R ¹ is hydrogen, fluorine or chlorine.

In some embodiments of compounds of Formula (I), Formula (IA) and (IB), each R ¹ is independently selected from hydrogen and halogen. Preferably, R1 is hydrogen, fluorine or chlorine ^; more preferably, ^R1 is hydrogen or fluorine.

In some embodiments of compounds of Formula (IC), each R ¹ is independently selected from hydrogen and halogen. Preferably, R1 is hydrogen, fluorine or chlorine ^; more preferably, ^R1 is hydrogen or fluorine.

In some embodiments of compounds of formula (I), ^formula (IA), (IB) and (IC), R is naphthyl optionally substituted with ^{1-3 R} , and each R is independently selected from F, Cl, Br, I, methyl, ethyl, isopropyl, ethynyl and hydroxyl, preferably, each ^R3 is independently selected from F, Cl, ethynyl and hydroxyl. Preferably, R is selected from ⁸ -Cl-naphthalen-1-yl, 3-OH-naphthalen-1-yl, 8-ethynyl-7-F-3-OH-naphthalen-1-yl and 8-ethynyl -7-F-Naphthalen-1-yl.

In some embodiments of compounds of formula (I), ^formula (IA) and (IB), R is naphthyl optionally substituted with ^{1-3 R} , and each R is independently selected from F, Cl, Br, I, methyl, ethyl, isopropyl, ethynyl and hydroxyl, preferably, each ^R3 is independently selected from F, Cl, ethynyl and hydroxyl. Preferably, R is selected from ⁸ -Cl-naphthalen-1-yl, 3-OH-naphthalen-1-yl, 8-ethynyl-7-F-3-OH-naphthalen-1-yl and 8-ethynyl -7-F-naphthalen-1-yl; more preferably, R is ⁸ -Cl-naphthalen-1-yl or 3-OH-naphthalen-1-yl.

In some embodiments of compounds of formula (IC), R ² is naphthyl optionally substituted with 1-3 R ³ , and each R ³ is independently selected from F, Cl, Br, I, methyl, Ethyl, isopropyl, ethynyl and hydroxyl, preferably, each ^R3 is independently selected from F, Cl, ethynyl and hydroxyl. Preferably, R is ⁸ -Cl-naphthalen-1-yl, 3-OH-naphthalen-1-yl, 8-ethynyl-7-F-3-OH-naphthalen-1-yl or 8-ethynyl- 7-F-naphthalen-1-yl; more preferably R2 is ⁸ -ethynyl-7-F-3-OH-naphthalen-1-yl.

In some embodiments of compounds of Formula (I), Formula (IA), (IB) and (IC), L ¹ is -O-.

In some embodiments of compounds of Formula (I), Formula (IA) and (IB), L ¹ is -O-.

In some embodiments of compounds of Formula (IC), L ¹ is -O-.

In some embodiments of the compounds of formula (I), formula (IA), (IB) and (IC), L ² is C _1-6 alkylene, preferably -CH ₂ - or -CH ₂ -CH ₂ -.

In some embodiments of the compounds of formula (I), formula (IA) and (IB), L ² is C _1-6 alkylene, preferably -CH ₂ - or -CH ₂ -CH ₂ -, more preferably is -CH ₂ -.

In some embodiments of the compound of formula (IC), L ² is C _1-6 alkylene, preferably -CH ₂ - or -CH ₂ -CH ₂ -, more preferably -CH ₂ -CH ₂ -.

In some embodiments of compounds of formula (I), formula (IA), (IB) and (IC), L ³ is a 3-6 membered heterocyclylene containing one N atom, preferably containing one N atom 5-6 membered heterocyclylene, preferably

更优选为

In some embodiments of compounds of formula (I), formula (IA) and (IB), L ³ is a 3-6 membered heterocyclylene containing one N atom, preferably a 5-6 membered heterocyclylene containing one N atom Heterocyclylene, preferably

more preferably

更优选为

In some embodiments of the compound of formula (IC), L ³ is a 3-6 membered heterocyclylene containing one N atom, preferably a 5-6 membered heterocyclylene containing one N atom, preferably

more preferably

In some embodiments of compounds of ^formula (I), ^formula (IA), (IB) and (IC), L and L are each independently selected from a covalent bond, C _1-6 alkylene, -OC _1-6 alkylene- and C _1-6 alkylene-O-. Preferably, -L ⁴ -L ⁵ - is -CH ₂ -CH ₂ -CH ₂ -O-CH ₂ -CH ₂ -, -CH ₂ -CH ₂ -CH ₂ -CH ₂ -, -CH ₂ -CH _{2 -CH2} - _CH2 -CH2-, _-CH2 - _CH2 - _CH2 - _CH2 - _{CH2 -} CH2-, _{-CH2 -} O _- CH2-, or _-CH2- .

In some embodiments of compounds of ^formula (I), ^formula (IA) and (IB), L and L are each independently selected from a covalent bond, C _1-6 alkylene, -OC _1-6 alkylene Alkyl- and C _1-6 alkylene-O-. Preferably, -L ⁴ -L ⁵ - is -CH ₂ -CH ₂ -CH ₂ -O-CH ₂ -CH ₂ -, -CH ₂ -CH ₂ -CH ₂ -CH ₂ -, -CH ₂ -CH _{2 -CH2} - _CH2 -CH2-, _-CH2 - _CH2 - _CH2 - _CH2 - _{CH2 -} CH2-, _{-CH2 -} O _- CH2-, or _-CH2- . Preferably, -L ⁴ -L ⁵ - is -CH ₂ -CH ₂ -CH ₂ -O-CH ₂ -CH ₂ -.

^In some embodiments of compounds of ^formula (IC), L and L are each independently selected from a covalent bond, C _1-6 alkylene, -OC _1-6 alkylene-, and C _1-6 alkylene Alkyl-O-. Preferably, -L ⁴ -L ⁵ - is -CH ₂ -CH ₂ -CH ₂ -O-CH ₂ -CH ₂ -, -CH ₂ -CH ₂ -CH ₂ -CH ₂ -, -CH ₂ -CH _{2 -CH2} - _CH2 -CH2-, _-CH2 - _CH2 - _CH2 - _CH2 - _{CH2 -} CH2-, _{-CH2 -} O _- CH2-, or _-CH2- . Preferably, -L ⁴ -L ⁵ - is -CH ₂ -O-CH ₂ - or -CH ₂ -.

In some embodiments of compounds of Formula (I), Formula (IA), (IB) and (IC), L6 is -C(O) ^- or -O-.

In some embodiments of compounds of formula (I), formula (IA) and (IB), L ⁶ is -C(O)- or -O-, preferably -C(O)-.

In some embodiments of compounds of Formula (IC), L ⁶ is -C(O)- or -O-.

In some embodiments of compounds of formula (I), formula (IA), (IB) and (IC), L is selected from

In some embodiments of compounds of formula (I), formula (IA), (IB) and (IC), ^A is -NH-; R is C _1-6 alkyl, preferably tert-butyl; and R ⁵ and ^R6 are both hydrogen.

In some embodiments of compounds of formula (I), formula (IA) and (IB), A is -NH-; R ⁴ is C _1-6 alkyl, preferably tert-butyl; and R ⁵ and R ⁶ Both are hydrogen.

In some embodiments of compounds of formula (IC), A is -NH-; R ⁴ is C _1-6 alkyl, preferably tert-butyl; and R ⁵ and R ⁶ are both hydrogen.

In some embodiments of compounds of formula (I), formula (IA), (IB) and (IC), A is -NH-; R ⁴ is C _1-6 alkyl, preferably tert-butyl; R ⁵ is C _1-6 alkyl, preferably methyl; and R ⁶ is hydrogen.

R ⁴为C _1-6烷基，优选为异丙基；R ⁵为C _1-6烷基，优选为甲基；且R ⁶为氢。 In some embodiments of compounds of formula (I), formula (IA), (IB) and (IC), A is 5-10 membered heteroarylene, preferably 5-6 membered heteroarylene, preferably

R ⁴ is C _1-6 alkyl, preferably isopropyl; R ⁵ is C _1-6 alkyl, preferably methyl; and R ⁶ is hydrogen.

In some embodiments of compounds of formula (I), formula (IA), (IB) and (IC), M is selected from the following:

In the above-mentioned formula (I), formula (IA), (IB) and (IC) of the present invention, the groups of all embodiments can be selected in any combination as appropriate, so as to obtain different general formula ranges or specific schemes. These ranges and schemes all belong to the present invention.

The present invention encompasses compounds resulting from any combination of the various embodiments.

In a particularly preferred embodiment, the compound of formula (I) is selected from:

Preparation

The compound of formula (IA) of the present invention can be prepared by the method shown in scheme I as follows:

Route 1

in

R ¹ , R ² , n, L and M are as defined above;

LG ^1a , LG ^2a , LG ^3a and X each independently represent a leaving group such as halogen, triflate, thiomethyl, methylsulfoxide, methylsulfone, methoxy or ethoxy ;

PG ^1a and PG ^2a each independently represent a protecting group, such as benzyloxycarbonyl (Cbz), tert-butoxycarbonyl (Boc), methyl or benzyl;

The method comprises the steps of:

1) reacting compound IA-1 with compound IA-2 to obtain compound IA-3;

2) reacting compound IA-3 with compound IA-4 to obtain compound IA-5;

3) subjecting compound IA-5 to a deprotection reaction to obtain compound IA-6;

4) performing a coupling reaction between compound IA-6 and compound IA-7 to obtain compound IA-8;

5) reacting Compound IA-8 with Compound IA-9 to obtain Compound IA-10; and

6) Subjecting compound IA-10 to a deprotection reaction to obtain compound IA.

The reaction of the above step (1) is preferably carried out in a suitable organic solvent and in the presence of a suitable organic base. The organic solvent can be selected from N,N-dimethylformamide, dimethyl sulfoxide, N,N-dimethylacetamide, tetrahydrofuran, 1,4-dioxane and any combination thereof, preferably N , N-dimethylacetamide. 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 0-80°C for 2-24 hours.

The reaction of the above step (2) is preferably carried out in a suitable organic solvent and in the presence of a metal catalyst, a ligand and a base. The metal catalyst may be a palladium metal catalyst, for example selected from tris(dibenzylideneacetone)dipalladium, [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride, triphenyl Palladium phosphine and palladium acetate, preferably [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride. The ligand may be a phosphorus ligand, for example selected from 4,5-bisdiphenylphosphine-9,9-dimethylxanthene, 2-dicyclohexylphosphine-2',6'-diisopropyl Oxy-1,1'-biphenyl, 2-dicyclohexylphosphine-2',4',6'-triisopropyl-1,1'-biphenyl and 2-dicyclohexylphosphine-2', 6'-dimethoxy-1,1'-biphenyl, preferably 4,5-bisdiphenylphosphine-9,9-dimethylxanthene. The base may be an organic base or an inorganic base, for example selected from N,N-diisopropylethylamine, triethylamine, sodium tert-butoxide, potassium carbonate, cesium carbonate and sodium carbonate, preferably cesium carbonate. The organic 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 For 1,4-dioxane. The reaction is preferably carried out at a temperature of 50-120°C for 2-16 hours.

The deprotection reaction of the above step (3) is preferably carried out in a suitable organic solvent and in the presence of a suitable acid. The organic solvent may be selected from ethyl acetate, methanol, 1,4-dioxane, dichloromethane and any combination thereof, preferably dichloromethane. The acid may be selected from hydrochloric acid, hydrobromic acid, p-toluenesulfonic acid, methanesulfonic acid and trifluoroacetic acid, preferably trifluoroacetic acid. The reaction is preferably carried out at a temperature of 0-40°C for 2-12 hours.

The coupling reaction of the above step (4) is preferably carried out in a suitable organic solvent and in the presence of a metal catalyst, a ligand and a base. The metal catalyst may be a palladium metal catalyst, such as tris(dibenzylideneacetone)dipalladium, [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride, triphenylphosphine palladium and palladium acetate, preferably tris(dibenzylideneacetone)dipalladium. The ligand may be a phosphorus ligand, for example selected from 4,5-bisdiphenylphosphine-9,9-dimethylxanthene, 2-dicyclohexylphosphine-2',6'-diisopropyl Oxy-1,1'-biphenyl, 2-dicyclohexylphosphine-2',4',6'-triisopropyl-1,1'-biphenyl, 2-dicyclohexylphosphine-2', 6'-dimethoxy-1,1'-biphenyl, preferably 2-dicyclohexylphosphine-2',6'-diisopropoxy-1,1'-biphenyl. The base may be an organic base or an inorganic base, for example selected from sodium tert-butoxide, potassium carbonate, cesium carbonate and sodium carbonate, preferably cesium carbonate. The organic 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 For 1,4-dioxane. The reaction is preferably carried out at a temperature of 50-120°C for 2-16 hours.

The reaction of the above step (5) is preferably carried out in a suitable organic solvent and in the presence of a suitable organic base and a condensing agent. The organic solvent may be selected from dichloromethane, tetrahydrofuran, acetonitrile, N,N-dimethylformamide and any combination thereof, preferably N,N-dimethylformamide. The organic base can be selected from N,N-diisopropylethylamine, triethylamine, 1,8-diazacyclo[5,4,0]undec-7-ene, preferably triethylamine . The condensing agent can be selected from 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate, 1-hydroxybenzotriazole, 1- (3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride, N-hydroxy-7-azabenzotriazole, dicyclohexylcarbodiimide and any combination thereof, preferably 2-(7-Azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate. The reaction is preferably carried out at a temperature of 0-60°C for 1-12 hours.

The deprotection reaction of the above step (6) is preferably carried out in a suitable organic solvent and in the presence of a suitable deprotection reagent. The organic solvent may be selected from ethyl acetate, methanol, acetonitrile, 1,4-dioxane, dichloromethane and any combination thereof, preferably dichloromethane. The deprotection reagent may be selected from hydrochloric acid, hydrobromic acid, p-toluenesulfonic acid, methanesulfonic acid, trifluoroacetic acid, aluminum trichloride and iodotrimethylsilane, preferably trimethyliodosilane. The reaction is preferably carried out at a temperature of 0-80°C for 1-12 hours.

The compound of formula (IB) of the present invention can be prepared by the method shown in Scheme 2 below:

route 2

in

wherein R ¹ , R ² , L and M are as defined above;

LG ^b , LG ^1b , LG ^2b , LG ^3b and LG ^4b each independently represent a leaving group such as halogen, triflate, thiomethyl, methylsulfoxide, methylsulfone, boronic acid, boron ester, tributyltin, methoxy or ethoxy;

PG ^1b represents a protecting group, such as benzyloxycarbonyl (Cbz) or tert-butoxycarbonyl (Boc);

The method comprises the steps of:

1) reacting compound IB-1 with compound IB-2 to obtain compound IB-3;

2) reacting compound IB-3 with compound IB-4 to obtain compound IB-5;

3) make compound IB-5 and compound IB-6 carry out coupling reaction to obtain compound IB-7;

4) reacting compound IB-7 with compound IB-8 to obtain compound IB-9; and

5) Subjecting compound IB-9 to a deprotection reaction to obtain compound IB.

The reaction of the above step (1) is preferably carried out in a suitable organic solvent and in the presence of a suitable organic base. The organic solvent can be selected from N,N-dimethylformamide, dimethyl sulfoxide, N,N-dimethylacetamide, tetrahydrofuran, 1,4-dioxane and any combination thereof, preferably N , N-dimethylacetamide. 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.

The reaction of the above step (2) is preferably carried out in a suitable organic solvent and in the presence of a suitable organic base. The organic solvent can be selected from acetonitrile, N,N-dimethylformamide, dimethyl sulfoxide, N,N-dimethylacetamide, tetrahydrofuran, 1,4-dioxane and any combination thereof, preferably to acetonitrile. 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 20°C-100°C for 2-48 hours.

The coupling reaction of the above step (3) is preferably carried out in a suitable solvent and in the presence of a metal catalyst and a base. The metal catalyst may be a palladium metal catalyst, such as tris(dibenzylideneacetone)dipalladium, [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride, methanesulfonic acid [n- Butylbis(1-adamantyl)phosphine](2-amino-1,1'-biphenyl-2-yl)palladium(II), tetrakistriphenylphosphinepalladium and palladium acetate, preferably [1,1 '-bis(diphenylphosphino)ferrocene]palladium dichloride or [n-butylbis(1-adamantyl)phosphine](2-amino-1,1'-biphenyl-2 -yl) palladium(II). The base may 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 cesium carbonate or 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 Combination of 1,4-dioxane and water. The reaction is preferably carried out at a temperature of 50-120°C for 2-16 hours.

The reaction of the above step (4) is preferably carried out in a suitable organic solvent and in the presence of a suitable organic base and a condensing agent. The organic solvent may be selected from dichloromethane, tetrahydrofuran, acetonitrile, N,N-dimethylformamide and any combination thereof, preferably N,N-dimethylformamide. The organic base can be selected from N,N-diisopropylethylamine, triethylamine, 1,8-diazacyclo[5,4,0]undec-7-ene, preferably N,N - diisopropylethylamine. The condensing agent can be selected from 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate, 1-hydroxybenzotriazole, 1- (3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride, N-hydroxy-7-azabenzotriazole, dicyclohexylcarbodiimide and any combination thereof, preferably 2-(7-Azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate. The reaction is preferably carried out at a temperature of 0-60°C for 1-12 hours.

The deprotection reaction of the above step (5) is preferably carried out in a suitable organic solvent and in the presence of a suitable deprotection reagent. The organic solvent may be selected from ethyl acetate, methanol, acetonitrile, 1,4-dioxane, dichloromethane and any combination thereof, preferably dichloromethane. The deprotection reagent may be selected from hydrochloric acid, hydrobromic acid, p-toluenesulfonic acid, methanesulfonic acid, trifluoroacetic acid, aluminum trichloride and iodotrimethylsilane, preferably trimethyliodosilane. The reaction is preferably carried out at a temperature of 0-80°C for 1-12 hours.

The compound of formula (IB) of the present invention can also be prepared by the method shown in Scheme 3 below:

Route 3

in

wherein R ¹ , R ² , L, L ¹ , L ² , L ³ , L ⁴ , L ⁵ , L ⁶ and M are as defined above;

LG ^b , LG ^1b , LG ^2b and LG ^4b each independently represent a leaving group such as halogen, triflate, thiomethyl, methylsulfoxide, methylsulfone, boronic acid, borate ester, Tributyltin, methoxy or ethoxy;

PG ^1b represents a protecting group, such as benzyloxycarbonyl (Cbz) or tert-butoxycarbonyl (Boc);

The method comprises the steps of:

1) reacting compound IB-1 with compound IB-2 to obtain compound IB-3;

2) reacting compound IB-3 with compound IB-10 to obtain compound IB-11;

3) performing a coupling reaction between compound IB-11 and compound IB-6 to obtain compound IB-12;

4) subjecting compound IB-12 to an oxidation reaction to obtain compound IB-13;

5) reacting compound IB-13 with compound IB-14 to obtain compound IB-15; and

6) Subjecting compound IB-15 to a deprotection reaction to obtain compound IB.

The reaction of the above step (1) is preferably carried out in a suitable organic solvent and in the presence of a suitable organic base. The organic solvent can be selected from N,N-dimethylformamide, dimethyl sulfoxide, N,N-dimethylacetamide, tetrahydrofuran, 1,4-dioxane and any combination thereof, preferably N , N-dimethylacetamide. 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.

The reaction of the above step (2) is preferably carried out in a suitable organic solvent and in the presence of a suitable organic base. The organic solvent can be selected from acetonitrile, N,N-dimethylformamide, dimethyl sulfoxide, N,N-dimethylacetamide, tetrahydrofuran, 1,4-dioxane and any combination thereof, preferably to acetonitrile. 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 20°C-100°C for 2-48 hours.

The coupling reaction of the above step (3) is preferably carried out in a suitable solvent and in the presence of a metal catalyst and a base. The metal catalyst may be a palladium metal catalyst, such as tris(dibenzylideneacetone)dipalladium, [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride, methanesulfonic acid [n- Butylbis(1-adamantyl)phosphine](2-amino-1,1'-biphenyl-2-yl)palladium(II), tetrakistriphenylphosphinepalladium and palladium acetate, preferably [1,1 '-bis(diphenylphosphino)ferrocene]palladium dichloride or [n-butylbis(1-adamantyl)phosphine](2-amino-1,1'-biphenyl-2 -yl) palladium(II). The base may 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 cesium carbonate or 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 Combination of 1,4-dioxane and water. The reaction is preferably carried out at a temperature of 50-120°C for 2-16 hours.

The reaction of the above step (4) 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 dichloromethane, tetrahydrofuran, acetonitrile, N,N-dimethylformamide and any combination thereof, preferably dichloromethane.

The oxidizing agent may be selected from Dess-Martin oxidizing agent, 2-iodylbenzoic acid, pyridinium dichromate and pyridinium chlorochromate, preferably Dess-Martin oxidizing agent. The reaction is preferably carried out at a temperature of 0-60°C for 1-16 hours.

The reaction of the above step (5) is preferably carried out in a suitable organic solvent and in the presence of a suitable base and reducing agent. The organic solvent can be selected from dichloromethane, tetrahydrofuran, acetonitrile, methanol, isopropanol and any combination thereof, preferably dichloromethane and isopropanol. The base may be selected from N,N-diisopropylethylamine, triethylamine, sodium bicarbonate and sodium acetate trihydrate, preferably sodium acetate trihydrate. The reducing agent may be selected from sodium borohydride, sodium cyanoborohydride, sodium triacetylborohydride and 2-picoline borane, preferably 2-picoline borane. The reaction is preferably carried out at a temperature of 0-60°C for 1-12 hours.

The deprotection reaction of the above step (6) is preferably carried out in a suitable organic solvent and in the presence of a suitable deprotection reagent. The organic solvent may be selected from ethyl acetate, methanol, acetonitrile, 1,4-dioxane, dichloromethane and any combination thereof, preferably dichloromethane. The deprotecting reagent may be selected from hydrochloric acid, hydrobromic acid, p-toluenesulfonic acid, methanesulfonic acid, trifluoroacetic acid, aluminum trichloride and iodotrimethylsilane, preferably trifluoroacetic acid. The reaction is preferably carried out at a temperature of 0-80°C for 1-12 hours.

The compound of formula (IC) of the present invention can be prepared by the method shown in Scheme 4 below:

Route 4

in

wherein R ¹ , R ² , L, L ¹ , L ² , L ³ , L ⁴ , L ⁵ , L ⁶ , n and M are as defined above;

LG ^c , LG ^1c , LG ^2c and LG ^3c each independently represent a leaving group such as halogen, triflate, thiomethyl, methylsulfoxide, methylsulfone, boronic acid, borate ester, Tributyltin, methoxy or ethoxy;

PG ^1c represents a protecting group, such as benzyloxycarbonyl (Cbz) or tert-butoxycarbonyl (Boc);

The method comprises the steps of:

1) reacting compound IC-1 with compound IC-2 to obtain compound IC-3;

2) reacting compound IC-3 with compound IC-4 to obtain compound IC-5;

3) performing a coupling reaction between compound IC-5 and compound IC-6 to obtain compound IC-7;

4) subjecting compound IC-7 to an oxidation reaction to obtain compound IC-8;

5) reacting compound IC-8 with compound IC-9 to obtain compound IC-10; and

6) Subjecting compound IC-10 to a deprotection reaction to obtain compound IC.

The reaction of the above step (1) is preferably carried out in a suitable organic solvent and in the presence of a suitable organic base. The organic solvent can be selected from N,N-dimethylformamide, dimethyl sulfoxide, N,N-dimethylacetamide, tetrahydrofuran, 1,4-dioxane and any combination thereof, preferably N , N-dimethylacetamide. 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.

The reaction of the above step (2) is preferably carried out in a suitable organic solvent and in the presence of a suitable organic base. The organic solvent can be selected from acetonitrile, N,N-dimethylformamide, dimethyl sulfoxide, N,N-dimethylacetamide, tetrahydrofuran, 1,4-dioxane and any combination thereof, preferably to acetonitrile. 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 20°C-100°C for 2-48 hours.

The coupling reaction of the above step (3) is preferably carried out in a suitable solvent and in the presence of a metal catalyst and a base. The metal catalyst may be a palladium metal catalyst, such as tris(dibenzylideneacetone)dipalladium, [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride, methanesulfonic acid [n- Butylbis(1-adamantyl)phosphine](2-amino-1,1'-biphenyl-2-yl)palladium(II), tetrakistriphenylphosphinepalladium and palladium acetate, preferably [1,1 '-bis(diphenylphosphino)ferrocene]palladium dichloride or [n-butylbis(1-adamantyl)phosphine](2-amino-1,1'-biphenyl-2 -yl) palladium(II). The base may 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 cesium carbonate or 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 Combination of 1,4-dioxane and water. The reaction is preferably carried out at a temperature of 50-120°C for 2-16 hours.

The reaction of the above step (4) 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 dichloromethane, tetrahydrofuran, acetonitrile, N,N-dimethylformamide and any combination thereof, preferably dichloromethane.

The oxidizing agent may be selected from Dess-Martin oxidizing agent, 2-iodylbenzoic acid, pyridinium dichromate and pyridinium chlorochromate, preferably Dess-Martin oxidizing agent. The reaction is preferably carried out at a temperature of 0-60°C for 1-16 hours.

The reaction of the above step (5) is preferably carried out in a suitable organic solvent and in the presence of a suitable base and reducing agent. The organic solvent can be selected from dichloromethane, tetrahydrofuran, acetonitrile, methanol, isopropanol and any combination thereof, preferably dichloromethane and isopropanol. The base may be selected from N,N-diisopropylethylamine, triethylamine, sodium bicarbonate and sodium acetate trihydrate, preferably sodium acetate trihydrate. The reducing agent may be selected from sodium borohydride, sodium cyanoborohydride, sodium triacetylborohydride and 2-picoline borane, preferably 2-picoline borane. The reaction is preferably carried out at a temperature of 0-60°C for 1-12 hours.

The deprotection reaction of the above step (6) is preferably carried out in a suitable organic solvent and in the presence of a suitable deprotection reagent. The organic solvent may be selected from ethyl acetate, methanol, acetonitrile, 1,4-dioxane, dichloromethane and any combination thereof, preferably dichloromethane. The deprotecting reagent may be selected from hydrochloric acid, hydrobromic acid, p-toluenesulfonic acid, methanesulfonic acid, trifluoroacetic acid, aluminum trichloride and iodotrimethylsilane, preferably trifluoroacetic acid. The reaction is preferably carried out at a temperature of 0-80°C for 1-12 hours.

Pharmaceutical compositions and kits

Another object of the present invention is to provide a pharmaceutical composition comprising a prophylactically and/or therapeutically effective amount of the compound of the present invention or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph Forms, solvates, N-oxides, isotope-labeled compounds, metabolites or prodrugs, and one or more pharmaceutically acceptable carriers.

Another object of the present invention is to provide a kit comprising the compound of the present invention or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N- Oxides, isotope-labeled compounds, metabolites or prodrugs, or pharmaceutical compositions of the invention.

"Pharmaceutically acceptable carrier" in the present invention refers to a diluent, adjuvant, excipient or vehicle administered together with a therapeutic agent, and it is suitable for contacting human beings and/or Tissues from other animals without undue toxicity, irritation, allergic response or other problems or complications commensurate with a reasonable benefit/risk ratio.

Pharmaceutically acceptable carriers that may be used in the pharmaceutical compositions or formulations of the present 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, etc.

The pharmaceutical composition may, for example, be in the form of a solid preparation, a semi-solid preparation, a liquid preparation or a gaseous preparation or the like. The solid preparations are, for example, tablets, capsules, powders, granules or suppositories, and the liquid preparations are, for example, solutions, suspensions or injections. The composition can also be in the form of liposomes, microspheres and the like.

The pharmaceutical compositions of the invention may act systemically and/or locally. For this purpose, they may be administered by a suitable route, for example by injection (e.g. intravenous, intraarterial, subcutaneous, intraperitoneal, intramuscular injection, including instillation) or transdermally; or by oral, buccal, transdermal Nasally, transmucosally, topically, in the form of ophthalmic formulations or by inhalation.

The content or amount of the compound of the present invention in the pharmaceutical composition can 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, especially preferably 0.5-100 mg.

In some embodiments, the present invention provides a method for preparing the pharmaceutical composition of the present invention, which method comprises the compound of the present invention or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorphic Forms, solvates, N-oxides, isotope-labeled compounds, metabolites or prodrugs are combined with one or more pharmaceutically acceptable carriers.

Treatments and uses

Another object of the present invention is to provide the compound of the present invention or its pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotope-labeled compound , metabolites or prodrugs, or the pharmaceutical composition of the present invention, which is used for the prevention and/or treatment of related diseases mediated by KRAS G12D.

Another object of the present invention is to provide the compound of the present invention or its pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotope-labeled compound , metabolites or prodrugs or the pharmaceutical composition of the present invention in the preparation of medicines for the prevention and/or treatment of KRAS G12D-mediated related diseases.

Another object of the present invention is to provide a method for preventing and/or treating related diseases mediated by KRAS G12D, which includes administering a preventive or therapeutically effective amount of the compound of the present invention or a pharmaceutically acceptable dose thereof to an individual in need thereof. Salts, stereoisomers, tautomers, polymorphs, solvates, N-oxides, isotopically labeled compounds, metabolites or prodrugs, or pharmaceutical compositions of the invention.

According to some embodiments of the present invention, the related disease mediated by KRAS G12D is tumor or cancer.

The term "effective amount" as used herein refers to an amount sufficient to achieve a desired prophylactic or therapeutic effect, eg, an amount that achieves alleviation of one or more symptoms associated with the disease being treated.

Dosage regimens may be adjusted to provide the optimum desired response. For example, a single bolus may be administered, several divided doses may be administered over time or the dose may be proportionally reduced or increased as indicated by the exigencies of the therapeutic situation. It is to be noted that dosage values may vary with the type and severity of the condition to be alleviated and may comprise single or multiple doses. It is further understood that for any given individual, the specific dosing regimen will be adjusted over time according to the needs of the individual and the professional judgment of the person administering the composition or supervising the administration of the composition.

The amount of a compound of this invention administered will depend on the individual being treated, the severity of the disorder or condition, the rate of administration, disposition of the compound, and the judgment of the prescribing physician. Generally, the effective dosage is about 0.0001 to about 50 mg per kg body weight per day, for example about 0.01 to about 10 mg/kg/day (single or divided administration). For a 70 kg human, this would amount to about 0.007 mg/day to about 3500 mg/day, such as about 0.7 mg/day to about 700 mg/day. Dosage levels up to the lower limit of the foregoing range may be sufficient in some cases, while in other cases larger doses may still be employed without causing any deleterious side effects, provided that the larger dose is first administered. The dose is divided into several smaller doses to be administered throughout the day.

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

The term "treating" as used herein means reversing, alleviating or eliminating the disorder or condition to which such term applies or the progression of one or more symptoms of such disorder or condition.

"Individual" as used herein includes a human or non-human animal. Exemplary human subjects include human subjects suffering from a disease (eg, a disease described herein) (referred to as a patient) or normal subjects. "Non-human animals" in the present invention 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

Embodiments of the present invention are described in detail below in conjunction with examples. However, those skilled in the art will understand 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.

Those who do not indicate specific conditions in the examples are carried out according to conventional conditions or the conditions suggested by the manufacturer. The reagents or instruments used were not indicated by the manufacturer, and they were all commercially available conventional products.

The structures of the compounds were determined by nuclear magnetic resonance ( ¹ H NMR) and/or mass spectroscopy (MS).

The measuring instrument of ¹ H NMR is JEOL Eclipse 400 nuclear magnetic instrument, and the measuring solvent is deuterated methanol (CD ₃ OD), deuterated chloroform (CDCl ₃ ) or hexadeuteriodimethyl sulfoxide (DMSO-d ₆ ), internal standard For tetramethylsilane (TMS), chemical shifts (δ) are given in parts per million (ppm).

The measuring instrument of MS is Agilent (ESI) mass spectrometer, manufacturer: Agilent, model: Agilent 6120B.

Preparative high performance liquid chromatography conditions are as follows:

Instrument model: Agilent 1260; Chromatographic column: Waters SunFire Prep C18 OBD (19mm×150mm×5.0μm); Chromatographic column temperature: 25°C; Flow rate: 20.0mL/min; Detection wavelength: 214nm; Elution gradient: (0min: 10 %A, 90%B; 16.0min: 90%A, 10%B); mobile phase A: acetonitrile; mobile phase B: 0.05% aqueous formic acid.

The thin-layer chromatography silica gel plate (TLC) uses an aluminum plate (20×20cm) produced by Merck, and the specification used for the separation and purification of the thin-layer chromatography is GF 254 (1mm) produced in Yantai.

The monitoring of reaction adopts thin-layer chromatography (TLC) or LC-MS; The developer system that uses comprises: dichloromethane and methanol system, normal hexane and ethyl acetate system, and sherwood oil and ethyl acetate system, the volume of solvent The ratio is adjusted according to the polarity of the compound or by adding triethylamine or the like.

Column chromatography generally uses 200-300 mesh silica gel as a carrier. The eluent system includes: dichloromethane and methanol system, and petroleum ether and ethyl acetate system. The volume ratio of the solvent is adjusted according to the polarity of the compound, and can also be adjusted by adding a small amount of triethylamine.

Unless otherwise specified, the reaction temperature is room temperature (20°C-35°C).

The reagents used in the examples were purchased from companies such as Acros Organics, Aldrich Chemical Company, and Tepper Chemical.

The meanings of abbreviations appearing in this article are shown in the table below.

abbreviation meaning TLC TLC LC-MS Liquid Chromatography-Mass Spectrometry DMF N,N-Dimethylformamide Pd(dppf)Cl ₂ [1,1'-Bis(diphenylphosphino)ferrocene]palladium dichloride CD ₃ OD deuterated methanol CDCl ₃ deuterated chloroform DMSO-d ₆ Hexadeuteriodimethylsulfoxide TMS Tetramethylsilane NMR nuclear magnetic resonance MS mass spectrometry the s Single peak (singlet) d Doublet t triplet q Quartet dd double doublet m Multiplets br Broad peak (broad) J Coupling constant Hz hertz

Preparation of intermediates:

Intermediate Preparation Example 1: Preparation of Benzyl 3,8-diazabicyclo[3.2.1]octane-8-carboxylate

Step 1: Preparation of 8-benzyl-3-tert-butyl 3,8-diazabicyclo[3.2.1]octane-3,8-dicarboxylate

Dissolve tert-butyl 3,8-diazabicyclo[3.2.1]octane-3-carboxylate (5.0 g, 23.55 mmol) in dichloromethane (50 mL). Under nitrogen protection, N,N-diisopropylethylamine (9.22 g, 70.66 mmol) was added. The system was cooled to 0°C, benzyl chloroformate (6.34g, 35.33mmol) was added dropwise thereto, and stirred at 25°C for 4 hours, then washed with water, dried and concentrated to obtain the title compound (8.16g, yield: 100 %).

MS m/z (ESI): 347.2 [M+H] ⁺ .

The second step: preparation of benzyl 3,8-diazabicyclo[3.2.1]octane-8-carboxylate

8-Benzyl-3-tert-butyl 3,8-diazabicyclo[3.2.1]octane-3,8-dicarboxylate (8.16g, 23.52mmol) was dissolved in trifluoroacetic acid (40mL) and dichloromethane (80 mL). The system was stirred at 20° C. for 2 hours, concentrated, and sodium bicarbonate solution was added to make the aqueous phase pH>8. Extracted three times with ethyl acetate (50 mL), dried the organic phase, concentrated and slurried (eluent: petroleum ether/ethyl acetate=10/1, volume ratio) for 12 hours, then filtered and dried the filter cake to obtain the title compound ( 4.6 g, yield: 79%).

MS m/z (ESI): 247.1 [M+H] ⁺ .

Intermediate preparation example 2: Preparation of ethyl (S)-3-(3-(2-(hydroxymethyl)pyrrolidin-1-yl)propoxy)propionate

The first step: the preparation of 3-(3-chloropropoxy) ethyl propionate

Dissolve 3-chloropropanol (5.0g, 52.88mmol) in acetonitrile (50mL), add ethyl acrylate (5.82g, 58.18mmol) and N-benzyl-trimethylammonium hydroxide aqueous solution (265.0mg, 1.58 mmol), and the reaction solution was stirred at 40°C for 72 hours. Completion of the reaction was monitored by thin-layer silica gel plates. The organic phase was concentrated under reduced pressure, and purified by silica gel column chromatography (eluent: petroleum ether/ethyl acetate=10/1, volume ratio) to obtain the title compound (3.5 g, yield: 34.0%).

MS m/z (ESI): 195.1 [M+H] ⁺ .

¹ H-NMR (400M Hz, CDCl ₃ ): δ4.15(q, J=7.2Hz, 2H), 3.71(t, J=6.0Hz, 2H), 3.62(t, J=6.4Hz, 2H), 3.58(t, J=6.0Hz, 2H), 2.56(t, J=6.0Hz, 2H), 2.03-1.97(m, 2H), 1.26(t, J=7.2Hz, 3H).

The second step: the preparation of ethyl (S)-3-(3-(2-(hydroxymethyl)pyrrolidin-1-yl)propoxy)propionate

Dissolve ethyl 3-(3-chloropropoxy)propionate (3.5g, 18.00mmol) in N,N-dimethylacetamide (30mL), add L-prolinol (1.82g, 18.00mmol ) and tetrabutylammonium iodide (50mg, 0.14mmol), and the reaction solution was stirred at 50°C for 16 hours. The reaction solution was concentrated under reduced pressure, and purified by preparative high-performance liquid chromatography to obtain the title compound (1.36 g, yield: 29.1%).

MS m/z (ESI): 260.2 [M+H].

¹ H-NMR (400M Hz, CDCl ₃ ): δ4.14(q, J=7.2Hz, 2H), 3.92-3.85(m, 2H), 3.82-3.75(m, 2H), 3.69-3.65(m, 2H), 3.55-3.48(m, 4H), 3.12-2.92(m, 2H), 2.54(t, J=6.4Hz, 2H), 2.17-2.04(m, 4H), 2.01-1.88(m, 2H) , 1.26 (t, J=7.2Hz, 3H).

Intermediate Preparation 3: 3-(2-(((S)-1-(3-(3-ethoxy-3-oxopropoxy)propyl)pyrrolidin-2-yl)methoxy )- Preparation of Benzyl 5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate

The first step: 4-(8-((benzyloxy)carbonyl)-3,8-diazabicyclo[3.2.1]octane-3-yl)-2-chloro-5,6-dihydropyridine Preparation of tert-butyl a[3,4-d]pyrimidine-7(8H)-carboxylate

2,4-dichloro-5,6-dihydropyrido[3,4-d]pyrimidine-7(8H)-tert-butyl carboxylate (1.0g, 3.29mmol) and N,N-diisopropyl Ethylamine (1.29 g, 9.87 mmol) was dissolved in DMF (10 mL), and benzyl 3,8-diazabicyclo[3.2.1]octane-8-carboxylate (0.85 g, 3.29 mmol) was added. The system was stirred at 80° C. for 2 hours, diluted with ethyl acetate (30 mL), washed with water, dried, concentrated and purified by silica gel column chromatography (eluent: petroleum ether/ethyl acetate=2/1, volume ratio ) to obtain the title compound (1.2 g, yield: 67%).

MS m/z (ESI): 514.2 [M+H] ⁺ .

The second step: 4-(8-((benzyloxy)carbonyl)-3,8-diazabicyclo[3.2.1]octane-3-yl)-2-(((S)-1-( 3-(3-ethoxy-3-oxopropoxy)propyl)pyrrolidin-2-yl)methoxy)-5,6-dihydropyrido[3,4-d]pyrimidine-7 Preparation of (8H)-tert-Butyl Carboxylate

4-(8-((Benzyloxy)carbonyl)-3,8-diazabicyclo[3.2.1]octane-3-yl)-2-chloro-5,6-dihydropyrido[3 ,4-d]pyrimidine-7(8H)-tert-butyl carboxylate (1.3g, 2.53mmol) and (S)-3-(3-(2-(hydroxymethyl)pyrrolidin-1-yl)propane Ethyloxy)propionate (1.31 g, 5.06 mmol) was dissolved in 1.4-dioxane (20 mL), and cesium carbonate (2.5 g, 7.59 mmol) was added. Nitrogen was replaced three times, and then [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (188.83mg, 0.25mmol) and 4,5-bisdiphenylphosphine-9,9- Dimethylxanthene (308.08mg, 0.51mmol), and stirred at 100°C for 12 hours. The system was cooled to room temperature, water (40 mL) was added, and extracted three times with ethyl acetate (30 mL), the organic layer was dried, concentrated and purified by silica gel column chromatography (eluent: dichloromethane/methanol=10/1, volume ratio ) to obtain the title compound (1.0 g, yield: 53.6%).

MS m/z (ESI): 737.2 [M+H] ⁺ .

The third step: 3-(2-(((S)-1-(3-(3-ethoxy-3-oxopropoxy)propyl)pyrrolidin-2-yl)methoxy)- Preparation of Benzyl 5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate

4-(8-((Benzyloxy)carbonyl)-3,8-diazabicyclo[3.2.1]octane-3-yl)-2-(((S)-1-(3-( 3-ethoxy-3-oxopropoxy)propyl)pyrrolidin-2-yl)methoxy)-5,6-dihydropyrido[3,4-d]pyrimidine-7(8H) - Tert-butyl carboxylate (1.0 g, 1.36 mmol) was dissolved in dichloromethane (20 mL), trifluoroacetic acid (10 mL) was added dropwise, and stirred at room temperature for 2 hours. The system was concentrated, and sodium bicarbonate solution was added to make the aqueous phase pH>8, then extracted three times with ethyl acetate (30 mL), the organic phase was dried and concentrated to obtain the title compound (0.66 g, yield: 76%).

MS m/z (ESI): 637.1 [M+H] ⁺ .

Intermediate Preparation Example 4: Preparation of 2-(3-(benzyloxy)naphthalen-1-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborane

The first step: the preparation of 3-benzyloxy-1-bromonaphthalene

4-Bronaphthalene-2-ol (5.0 g, 22.19 mmol) and potassium carbonate (9.20 g, 66.57 mmol) were sequentially added to acetonitrile (100 mL), and benzyl bromide (4.22 g, 24.41 mmol) was added dropwise. After dropping, the system was stirred at 80°C for 1 hour. The system was cooled to room temperature, and acetonitrile was removed by rotary evaporation under reduced pressure. The crude product was dissolved in ethyl acetate (150 mL), washed twice with saturated brine (50 mL), the organic phase was dried over anhydrous sodium sulfate, concentrated and spin-dried to obtain the title compound (6.7 g, yield: 91.58%).

MS m/z (ESI): 313.1 [M+H] ⁺ .

¹ H-NMR (400MHz, CDCl ₃ ): δ8.15-8.12 (d, J = 8.0Hz, 1H), 7.71-7.69 (d, J = 8.0Hz, 1H), 7.75- 7.32 (m, 8H), 7.20-7.19 (d, J=2.0Hz, 1H), 5.17 (s, 2H).

The second step: the preparation of 2-(3-(benzyloxy)naphthalen-1-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborane

Under nitrogen protection, 3-benzyloxy-1-bromonaphthalene (500mg, 1.60mmol), biboronic acid pinacol ester (608.11mg, 2.39mmol), potassium acetate (313.37mg, 3.19mmol), Pd (dppf )Cl ₂ (58.41mg, 79.82umol) was added to toluene (10mL), and the system was stirred at 100°C overnight. The system was cooled to room temperature and purified by silica gel column chromatography (eluent: petroleum ether/ethyl acetate=30/1, volume ratio) to obtain the title compound (531 mg, yield: 92.33%).

MS m/z (ESI): 361.2 [M+H] ⁺ .

Intermediate Preparation Example 5: Preparation of (S)-tert-butyl 3-(3-(2-(hydroxymethyl)pyrrolidinyl-1-yl)propoxy)propionate

The first step: the preparation of 3-(3-chloropropoxy) tert-butyl propionate

Dissolve 3-chloropropanol (4.0g, 42.31mmol) in acetonitrile (50mL), add tert-butyl acrylate (5.0g, 39.01mmol) and N-benzyl-trimethylammonium hydroxide aqueous solution (195.7mg, 1.17mmol), and the reaction solution was stirred at 40°C for 48 hours. Completion of the reaction was monitored by thin-layer silica gel plates. The organic phase was concentrated under reduced pressure, and purified by silica gel column chromatography (eluent: petroleum ether/ethyl acetate=30/1, volume ratio) to obtain the title compound (4.0 g, yield: 46.5%).

MS m/z (ESI): 223.1 [M+H] ⁺ .

¹ H-NMR (400M Hz, CDCl ₃ ): δ3.67(t, J=6.4Hz, 2H), 3.62(t, J=6.4Hz, 2H), 3.57(t, J=6.0Hz, 2H), 2.47(t, J=6.4Hz, 2H), 2.03-1.97(m, 2H), 1.45(s, 9H).

The second step: preparation of (S)-3-(3--(2-(hydroxymethyl)pyrrolidin-1-yl)propoxy)propionate tert-butyl ester

Dissolve tert-butyl 3-(3-chloropropoxy)propionate (3.9g, 17.51mmol) in N,N-dimethylacetamide (20mL), add L-prolinol (1.77g, 17.51 mmol) and tetrabutylammonium iodide (15mg), and the reaction solution was stirred at 60°C for 16 hours. The reaction solution was concentrated under reduced pressure, and purified by preparative high performance liquid chromatography (acetonitrile/1‰TFA system) to obtain the title compound (1.65 g, yield: 76.4%).

MS m/z (ESI): 288.2 [M+H] ⁺ .

¹ H-NMR (400M Hz, CDCl ₃ ): δ10.14(s,1H),3.95-3.91(m,1H),3.86-3.81(m,1H),3.79-3.76(m,1H),3.69- 3.63(m,2H),3.59-3.54(m,4H),3.17-3.13(m,1H),3.06-2.98(m,1H),2.47(t,J＝6.0Hz,2H),2.19-2.07( m,4H), 1.95-1.89(m,2H), 1.45(s,9H).

Intermediate Preparation Example 6: 3-(2-(((S)-1-(3-(3-(tert-butoxy)-3-oxopropoxy)propyl)pyrrolidin-2-yl) Methoxy)-7- Preparation of Benzyl Chloro-8-fluoropyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate

The first step: Preparation of ethyl 4-amino-6-chloro-5-fluoropyridine-3-carboxylate

2-Chloro-3-fluoro-5-iodo-pyridin-4-amine (3.60 g, 13.08 mmol), triethylamine (1.99 g, 19.62 mmol) and Pd(dppf)Cl ₂ (478.59 mg, 654.07 μmol) Add absolute ethanol (100 mL) sequentially, and stir the reaction system at 80° C. overnight under a carbon monoxide atmosphere. The system was cooled to room temperature and purified by silica gel column chromatography (eluent: petroleum ether/ethyl acetate=4/1, volume ratio) to obtain the title compound (2.5 g, yield: 83.05%).

MS m/z (ESI): 219.0 [M+H] ⁺ .

The second step: Preparation of ethyl 6-chloro-5-fluoro-4-[(2,2,2-trichloroacetyl)carbamoylamino]pyridine-3-carboxylate

Add ethyl 4-amino-6-chloro-5-fluoropyridine-3-carboxylate (2.5 g, 10.99 mmol) into anhydrous tetrahydrofuran (50 mL), slowly add 2,2,2-trichloroacetyl isocyanate ( 3.14g, 16.49mmol), and stirred at 25°C for 15 minutes. The tetrahydrofuran was removed by rotary evaporation under reduced pressure, the crude product was added to methyl tert-butyl ether, stirred thoroughly for 5 minutes, filtered with suction, and the filter cake was dried to obtain the title compound (4.3 g, yield: 91.29%).

MS m/z (ESI): 405.9 [M+H] ⁺ .

The third step: the preparation of 7-chloro-8-fluoro-pyrido[4,3-d]pyrimidine-2,4-diol

Add ethyl 6-chloro-5-fluoro-4-[(2,2,2-trichloroacetyl)carbamoylamino]pyridine-3-carboxylate (4.30g, 10.04mmol) into anhydrous methanol (50mL ), ammonia/methanol (10 mL) was added dropwise, and stirred at 25° C. for 1.5 hours. Methanol was removed by rotary evaporation under reduced pressure, and the crude product was added to methyl tert-butyl ether, stirred thoroughly for 10 minutes, filtered with suction, and the filter cake was collected to obtain the title compound (2.7 g, yield: 96.99%).

MS m/z (ESI): 216.0 [M+H] ⁺ .

Step 4: Preparation of 2,4,7-trichloro-8-fluoropyrido[4,3-d]pyrimidine

Add N,N-diisopropylethylamine (1.16g, 9.01mmol) dropwise to phosphorus oxychloride (7mL) cooled in an ice-salt bath, and slowly add 7-chloro-8-fluoro-pyrido[ 4,3-d]pyrimidine-2,4-diol (500 mg, 1.80 mmol). After the addition was complete, the reaction system was stirred at 100°C for 1.5 hours. The system was cooled to room temperature, most of the phosphorus oxychloride was removed by rotary evaporation under reduced pressure, chloroform (10 mL) was added and the remaining phosphorus oxychloride was removed by rotary evaporation under reduced pressure to obtain the title compound (513 mg, yield: 98.50%) , which is used directly in the next step.

MS m/z (ESI): 251.9 [M+H] ⁺ .

The fifth step: 3-(2,7-dichloro-8-fluoropyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8 - Preparation of benzyl carboxylate

2,4,7-Trichloro-8-fluoropyrido[4,3-d]pyrimidine (513 mg, 1.78 mmol) was added into dichloromethane (15 mL), and the temperature was lowered to -40°C. At this temperature, 3,8-diazabicyclo[3.2.1]octane-8-carboxylate benzyl ester (481.06mg, 1.95mmol) was added, and N,N-diisopropylethylamine was slowly added dropwise (688.44mg, 5.33mmol), after dropping, keep stirring at this temperature for 10 minutes. Sodium bicarbonate solution (20 mL) was added, extracted three times with ethyl acetate (15 mL), and the organic layer was dried over anhydrous sodium sulfate, concentrated and spin-dried, and then purified by column chromatography (eluent: petroleum ether/ethyl acetate ester=3/1, volume ratio), to obtain the title compound (810 mg, yield: 88.81%)

MS m/z (ESI): 462.1 [M+H] ⁺ .

The sixth step: 3-(2-(((S)-1-(3-(3-(tert-butoxy)-3-oxopropoxy)propyl)pyrrolidin-2-yl)methoxy Preparation of benzyl)-7-chloro-8-fluoropyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate

3-(2,7-dichloro-8-fluoropyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid Benzyl ester (500mg, 973.39μmol), (S)-tert-butyl 3-(3-(2-(hydroxymethyl)pyrrolidinyl-1-yl)propoxy)propionate (1.23g, 2.92mmol) and N,N-diisopropylethylamine (629.01mg, 4.87mmol) were sequentially added to acetonitrile (20mL), and the system was stirred at 80°C for 20 hours. The system was cooled to room temperature, and purified by silica gel column chromatography (eluent: ethyl acetate) to obtain the title compound (197 mg, yield: 26.96%).

MS m/z (ESI): 713.3 [M+H] ⁺ .

Intermediate Preparation 7: 3-(2,7-dichloro-8-fluoropyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane -8-carboxy Preparation of tert-butyl acid

Using the synthetic route of intermediate preparation example 6, the reaction raw material 3,8-diazabicyclo[3.2.1]octane-8-benzyl carboxylate in the fifth step was replaced by 3,8-diazabicyclo[ 3.2.1] tert-butyl octane-8-carboxylate to obtain the title compound (3.5g, yield: 80.5%)

MS m/z (ESI): 428.2 [M+H] ⁺ .

Intermediate Preparation 8: 3-(7-chloro-2-(2-(4-((2-ethoxy-2-oxoethoxy)methyl)piperidin-1-yl)ethoxy )-8-Fluoropyridine Preparation of tert-butyl pyrimido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate

The first step: the preparation of tert-butyl 4-(2-ethoxy-2-oxoethoxy)methyl)piperidine-1-carboxylate

In a 100mL round-bottomed flask, N-tert-butoxycarbonyl-4-piperidinemethanol (3.0g, 13.95mmol), rhodium diacetate (0.31g, 0.7mmol) and dichloromethane (30mL) were added successively. The reaction mixture was cooled to 0°C, and ethyl diazoacetate (2.39 g, 20.93 mmol) was slowly added dropwise thereto in batches. The reaction mixture was warmed up to 25°C and reacted for 12 hours with stirring. The reaction mixture was poured into water, extracted three times with ethyl acetate (15 mL), the organic phase was washed twice with saturated brine (15 mL), the obtained organic phase was dried over anhydrous sodium sulfate, filtered, and dried, and the resulting residue was passed through a column Purification by chromatography (eluent: petroleum ether/ethyl acetate=9/1, volume ratio) gave the title compound (2.2 g, yield: 52.4%).

MS m/z (ESI): 302.2 [M+H] ⁺ .

The second step: the preparation of 2-(4-piperidinylmethoxy) ethyl acetate hydrochloride

To a 100mL round bottom flask, add 4-(2-ethoxy-2-oxoethoxy)methyl)piperidine-1-carboxylic acid tert-butyl ester (2.2g, 7.31mmol) and 4M hydrogen chloride 1,4-dioxane solution (10 mL), the reaction mixture was stirred at 25°C for 2 hours, and the solvent was distilled off under reduced pressure to obtain the title compound (1.73 g, yield: 100%).

MS m/z (ESI): 202.2 [M+H] ⁺ .

The third step: the preparation of ethyl 2-(1-(2-hydroxyethyl)piperidin-4-yl)methoxy)acetate

In a 100mL round bottom flask, add 2-(4-piperidinylmethoxy) ethyl acetate hydrochloride (1.5g, 6.31mmol), potassium carbonate (1.74g, 12.62mmol), acetonitrile (20mL) in sequence and 2-iodoethanol (1.63 g, 9.5 mmol), replaced with nitrogen three times, the reaction mixture was heated to 80° C., and reacted for 12 hours under stirring. The reaction mixture was cooled to room temperature, filtered, the filter residue was washed with a small amount of ethyl acetate, the obtained organic phase was spin-dried, and the obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate/methanol=10/1, volume ratio ) to obtain the title compound (1.25 g, yield: 80.8%).

MS m/z (ESI): 246.1 [M+H] ⁺ .

The fourth step: 3-(7-chloro-2-(2-(4-((2-ethoxy-2-oxoethoxy)methyl)piperidin-1-yl)ethoxy)- Preparation of tert-butyl 8-fluoropyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate

Into a 10mL microwave tube, sequentially add 3-(2,7-dichloro-8-fluoropyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1] Octane-8-carboxylic acid tert-butyl ester (200mg, 0.47mmol), 2-(1-(2-hydroxyethyl)piperidin-4-yl)methoxy)ethyl acetate (172mg, 0.71mmol), N,N-Diisopropylethylamine (242mg, 1.88mmol) and 1,4-dioxane (5mL), nitrogen gas was bubbled into the reaction mixture for 1 minute, and the resulting mixture was heated to 120°C by microwave, under stirring React for 5 hours. The solvent was evaporated under reduced pressure, and the obtained residue was purified by silica gel column chromatography (eluent: dichloromethane/methanol=10/1, volume ratio) to obtain the title compound (146 mg, yield: 48.9%).

MS m/z (ESI): 637.2 [M+H] ⁺ .

Intermediate Preparation Example 9: Preparation of (S)-5-(2-(hydroxymethyl)pyrrolidin-1-yl)ethyl pentanoate

Dissolve (S)-pyrrolidin-2-ylmethanol (505.05 mg, 4.94 mmol) in tetrahydrofuran (20 mL), and add ethyl 5-bromovalerate (1.04 g, 4.94 mmol) and triethylamine (504.32 mg , 4.94mmol), the reaction mixture was refluxed at 70°C for 2 hours. The mixture was cooled to room temperature, solids were removed by filtration, the filtrate was evaporated to dryness, the concentrate was diluted with water (20 mL), extracted three times with dichloromethane (10 mL), the organic phase was dried over anhydrous sodium sulfate, and the solvent was removed under reduced pressure to give the title Compound (900 mg, yield: 79.4%).

MS m/z (ESI): 230.2 [M+H] ⁺ .

Intermediate Preparation Example 10: Preparation of (S)-6-(2-(hydroxymethyl)pyrrolidin-1-yl)hexanoic acid ethyl ester

Using the synthetic route of Intermediate Preparation Example 9, replacing the raw material 5-bromovaleric acid ethyl ester with 6-bromohexanoic acid ethyl ester, the title compound (1.1 g, yield: 67.5%) was obtained.

MS m/z (ESI): 244.2 [M+H] ⁺ .

Intermediate Preparation Example 11: Preparation of (S)-7-(2-(hydroxymethyl)pyrrolidin-1-yl)ethyl heptanoate

Using the synthetic route of Intermediate Preparation Example 9, replacing the raw material 5-bromovaleric acid ethyl ester with 7-bromoheptanoic acid ethyl ester, the title compound (850 mg, yield: 75.5%) was obtained.

MS m/z (ESI): 258.2 [M+H] ⁺ .

Intermediate Preparation 12: (2S,4R)-4-Hydroxy-1-(3-methyl-2-(3-(piperidin-4-ylmethoxy)isoxazol-5-yl)butyryl )- Preparation of N-(S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide

The first step: the preparation of 2-(3-bromoisoxazol-5-yl)ethanol

To a stirred solution of 3-butyn-1-ol (28.5 g, 386.5 mmol) and potassium bicarbonate (29.0 g, 289.9 mmol) in ethyl acetate/water (120 mL/12 mL) was added dropwise at room temperature A solution of 1,1-dibromoformaldoxime (20.0 g, 96.6 mmol) in ethyl acetate (50 mL). The resulting solution was stirred at room temperature for 16 hours. It was then diluted with water, and the organic layer was separated and extracted three times with ethyl acetate (100 mL). The organic layer was washed with saturated brine (150 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: ethyl acetate/petroleum ether=2/1, volume ratio) to obtain the title compound (16.4 g, yield: 84.0%).

MS m/z (ESI): 192.0 [M+H] ⁺ .

The second step: the preparation of 2-(3-bromoisoxazol-5-yl)acetic acid

2-(3-Bromoisoxazol-5-yl)ethanol (16.4g, 85.4mmol) was dissolved in acetone (190mL), and Jones reagent (70mL) was added dropwise at 0°C, stirred at this temperature for 30 minutes and then removed to room temperature and stirred at room temperature for 16 hours. Add water (300 mL) to the reaction solution, extract three times with ethyl acetate (100 mL), wash the combined organic phase with saturated brine (150 mL), dry the organic phase with anhydrous sodium sulfate, and concentrate under reduced pressure to obtain the title compound ( 18 g, yield: 86.2%).

MS m/z (ESI): 206.0 [M+H] ⁺ .

The third step: the preparation of 2-(3-bromoisoxazol-5-yl) methyl acetate

2-(3-Bromoisoxazol-5-yl)acetic acid (18g, 87.8mmol) was dissolved in methanol (150mL), concentrated sulfuric acid (2mL) was added dropwise, and stirred at 70°C for 2.5 hours. Concentrate the reaction solution, add ethyl acetate (30 mL) to dilute the residue, wash with water (30 mL), collect the organic phase, wash the combined organic phase with saturated brine (50 mL), dry the organic phase with anhydrous sodium sulfate, and reduce Concentrate under pressure. The residue was purified by silica gel column chromatography (eluent: ethyl acetate/petroleum ether=4/1, volume ratio) to obtain the title compound (15 g, yield: 78%).

MS m/z (ESI): 220.0 [M+H] ⁺ .

The fourth step: the preparation of 2-(3-bromoisoxazol-5-yl)-3-methylbutanoic acid

2-(3-Bromoisoxazol-5-yl)methyl acetate (8.7g, 39.6mmol) was dissolved in tetrahydrofuran (90mL), the temperature was lowered to 0°C, and potassium tert-butoxide (5.88g, 52.59mmol), 2-iodopropane (12.75g, 75.12mmol) was added dropwise at low temperature, replaced with nitrogen three times, and then stirred and reacted at room temperature for 16 hours. The reaction solution was concentrated, the residue was adjusted to pH=3-4 with 1N hydrochloric acid solution, extracted twice with ethyl acetate (40 mL), the organic phase was washed with saturated brine (50 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: ethyl acetate/petroleum ether=2/1, volume ratio) to obtain the title compound (4.2 g, yield: 42.8%).

MS m/z (ESI): 248.0 [M+H] ⁺ .

Step 5: Preparation of 2-(3-methoxyisoxazol-5-yl)-3-methylbutanoic acid

2-(3-Bromoisoxazol-5-yl)-3-methylbutanoic acid (3.0g, 12.16mmol) was dissolved in methanol (40mL), potassium hydroxide (6.8g, 120.16mmol) was added, and the temperature was raised to 68°C, stirring for 4 hours. Adjust the pH to 3-4 with 1N hydrochloric acid solution, extract twice with ethyl acetate (20 mL), dry over anhydrous sodium sulfate, and concentrate. The residue was purified by reverse phase chromatography (mobile phase: acetonitrile: 0.1% aqueous formic acid, 0-40%) to obtain the title compound (1.2 g, yield: 49.6%).

MS m/z (ESI): 200.1 [M+H] ⁺ .

Step 6: Preparation of 2-(3-hydroxyisoxazol-5-yl)-3-methylbutanoic acid

Dissolve 2-(3-methoxyisoxazol-5-yl)-3-methylbutanoic acid (1g, 5.0mmol) in acetic acid (8mL), add hydrobromic acid aqueous solution (6mL), and heat to 60°C , stirred for 16 hours. The reaction solution was concentrated, and the residue was purified by reverse phase chromatography (mobile phase: acetonitrile: 0.1% aqueous formic acid, 0-40%) to obtain the title compound (0.7 g, yield: 75.3%).

MS m/z (ESI): 186.1 [M+H] ⁺ .

Step 7: Preparation of 2-(3-hydroxyisoxazol-5-yl)-3-methylbutyric acid methyl ester

2-(3-Hydroxyisoxazol-5-yl)-3-methylbutanoic acid (700mg, 3.76mmol) was dissolved in methanol (4mL), and thionyl chloride (1.92g, 16.16mmol) was added dropwise, in Stir at 25°C for 3 hours. After concentrating the reaction solution, add ethyl acetate (20 mL) to dilute the residue, wash with water (10 mL), wash the organic phase with saturated brine (10 mL), dry the organic phase with anhydrous sodium sulfate, and concentrate under reduced pressure to obtain the title compound (0.6 g, yield: 79.7%).

MS m/z (ESI): 200.1 [M+H]+.

The eighth step: 4-(5-(1-methoxy-3-methyl-1-oxobutane-2-yl)isoxazol-3-yl)oxy)methylpiperidine-1- Preparation of tert-butyl carboxylate

2-(3-Hydroxyisoxazol-5-yl)-3-methylbutanoic acid methyl ester (600mg, 2.86mmol) was dissolved in DMF (1mL), and 4-(bromomethyl)piperidine-1- Tert-butyl carboxylate (2.09g, 7.15mmol) and potassium carbonate (1.19g, 8.58mmol) were stirred at 50°C for 3 hours. The reaction solution was cooled to room temperature, water was added to the reaction solution, extracted three times with ethyl acetate (20 mL), the organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The title compound (1.0 g, yield: 79.3%) was obtained.

MS m/z (ESI): 397.2 [M+H] ⁺ .

Step 9: Preparation of 2-(3-(1-(tert-butoxycarbonyl)piperidin-4-yl)methoxy)isoxazol-5-yl)-3-methylbutanoic acid

4-(5-(1-methoxy-3-methyl-1-oxobutan-2-yl)isoxazol-3-yl)oxy)methylpiperidine-1-carboxylic acid tert Butyl ester (1 g, 1.26 mmol) was dissolved in methanol (10 mL) and water (5 mL), sodium hydroxide (151.3 mg, 3.79 mmol) was added, and stirred at room temperature for 2 hours. Concentrate, dilute with water, adjust pH=3~4 with 1N hydrochloric acid solution, extract three times with ethyl acetate (40mL), wash the organic phase with saturated brine (50mL), dry the organic phase with anhydrous sodium sulfate, and concentrate under reduced pressure , the residue was purified by silica gel column chromatography (eluent: dichloromethane/methanol=10/1, volume ratio) to obtain the title compound (0.25 g, yield: 50.8%).

MS m/z (ESI): 383.2 [M+H] ⁺ .

The tenth step: 4-((5-(1-(2S,4R)-4-hydroxy-2-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl Base) carbamoyl) pyrrolidin-1-yl)-3-methyl-1-oxobutane-2-yl)isoxazol-3-yl)oxy)methyl)piperidine-1-carboxy Preparation of tert-butyl acid

2-(3-(1-(tert-butoxycarbonyl)piperidin-4-yl)methoxy)isoxazol-5-yl)-3-methylbutanoic acid (115.39mg, 301μmol) and (2S ,4R)-4-Hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (99.6mg, 301μmol) Dissolve in DMF (2mL), cool down to 0°C, add N,N-diisopropylethylamine (111mg, 860μmol) and 2-(7-azobenzotriazole)-N,N,N',N '-Tetramethyluronium hexafluorophosphate (163 mg, 430 μmol). Stir at 0°C for 1 hour. Add water (10 mL) to the reaction solution, extract three times with ethyl acetate (20 mL), wash the organic phase with saturated brine (20 mL), dry the organic phase with anhydrous sodium sulfate, and concentrate under reduced pressure, and pass the residue through a silica gel column Purification by chromatography (eluent: dichloromethane/methanol=10/1, volume ratio) gave the title compound (100 mg, yield: 48.1%).

MS m/z (ESI): 696.3 [M+H] ⁺ .

The eleventh step: (2S,4R)-4-hydroxy-1-(3-methyl-2-(3-(piperidin-4-ylmethoxy)isoxazol-5-yl)butyryl) Preparation of -N-(S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide

4-((5-(1-(2S,4R)-4-hydroxy-2-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)amino Formyl)pyrrolidin-1-yl)-3-methyl-1-oxobutan-2-yl)isoxazol-3-yl)oxy)methyl)piperidine-1-carboxylic acid tert-butyl The ester (100 mg, 143.71 μmol) was dissolved in dichloromethane (2.00 mL), 1,4-dioxane hydrochloride (2.00 mL) was added, and stirred at room temperature for 1 hour. The reaction solution was concentrated to obtain the title compound (70 mg, yield: 81.8%).

MS m/z (ESI): 596.3 [M+H] ⁺ .

Intermediate Preparation 13: ((2-fluoro-8-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalen-1-yl )ethynyl) Preparation of Triisopropylsilane

Into a 50mL single-necked bottle, add 7-fluoro-8-(triisopropylsilyl)ethynyl)naphthalene-1-yltrifluoromethanesulfonic acid (200.00mg, 421.42μmol) and pinacol diborate in sequence (214.03mg, 842.84μmol), potassium acetate (124.08mg, 1.26mmol), 1,4-dioxane (5.0mL) and [1,1'-bis(diphenylphosphino)ferrocene]dichloro Palladium dichloromethane complex (172.15 mg, 210.71 μmol) was replaced with nitrogen three times, and the reaction mixture was stirred at 104°C for 12 hours. After the reaction solution was cooled to room temperature, water was added to the reaction solution, extracted twice with ethyl acetate (20 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: petroleum ether) to obtain the title compound (54 mg, yield: 28.32%) as a yellow solid.

MS m/z (ESI): 453.3 [M+H] ⁺ .

¹ H-NMR (400MHz, CDCl ₃ ) δ7.84-7.75(m, 3H), 7.42(dd, J=8.0, 8.0Hz, 1H), 7.27(d, J=8.0Hz, 1H), 7.24(d , J=4.0Hz, 1H), 1.42(d, J=8.0Hz, 12H), 1.25(s, 3H), 1.18-1.14(m, 18H).

Intermediate Preparation 14: 3-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalene- Preparation of tert-butyl 1-yl)-2-(2-oxoethoxy)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate

The first step: the preparation of 7-bromo-8-fluoro-1H-quinazoline-2,4-dione

2-Amino-4-bromo-3-fluorobenzoic acid (2.0g, 8.5mmol) and urea (2.7g, 45.1mmol) were mixed and heated to 200°C for 3 hours. After cooling to room temperature, ethyl acetate (200 mL) was added and stirred for 30 minutes. After filtration, the filter cake was collected and concentrated to dryness under reduced pressure to obtain the title compound (1.9 g, yield: 86.4%).

MS m/z (ESI): 260.0 [M+H] ⁺ .

The second step: the preparation of 7-bromo-2,4-dichloro-8-fluoroquinazoline

Dissolve 7-bromo-8-fluoro-1H-quinazoline-2,4-dione (1.9g, 7.3mmol) in N,N-diisopropylethylamine (6mL) and phosphorus oxychloride (40mL ), stirred at 110°C for 10 hours. After cooling to room temperature and concentrating, the residue was purified by silica gel column chromatography (eluent: ethyl acetate/petroleum ether=20/1, volume ratio) to obtain the title compound (1.6 g, yield: 73.7%).

MS m/z (ESI): 297.0 [M+H] ⁺ .

The third step: 3-(7-bromo-2-chloro-8-fluoro-quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl Preparation of esters

7-bromo-2,4-dichloro-8-fluoroquinazoline (1.6g, 5.4mmol) and tert-butyl 3,8-diazabicyclo[3.2.1]octane-8-carboxylate ( 1.2g, 5.7mmol) was dissolved in 1,4-dioxane (30mL), triethylamine (3mL) was added, and the reaction was stirred at 50°C for 30 minutes. Concentrate the reaction solution, add ethyl acetate (30 mL) to dilute the residue, wash with water (30 mL), collect the organic phase, wash the organic phase with saturated brine (50 mL), dry the organic phase with anhydrous sodium sulfate, and concentrate under reduced pressure . The residue was purified by silica gel column chromatography (eluent: ethyl acetate/petroleum ether=2/1, volume ratio) to obtain the title compound (2.2 g, yield: 86.3%).

MS m/z (ESI): 473.0 [M+H] ⁺ .

The fourth step: 3-(7-bromo-8-fluoro-2-(2-hydroxyethoxy)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane- Preparation of tert-butyl 8-carboxylate

Dissolve ethylene glycol (328.9mg, 5.3mmol) in DMF (5mL), lower the temperature to 0°C, slowly add sodium hydride (40.3mg, 1.0mmol) under nitrogen protection, keep stirring at low temperature for 30 minutes, add 3- (7-Bromo-2-chloro-8-fluoro-quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester (0.5g, 1.0 mmol), kept stirring at low temperature for 3 hours. The reaction solution was poured into water (40 mL), extracted twice with ethyl acetate (20 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: ethyl acetate/petroleum ether=1/1, volume ratio) to obtain the title compound (230.0 mg, yield: 43.6%).

MS m/z (ESI): 497.0 [M+H] ⁺ .

The fifth step: 3-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalene-1-yl) Preparation of tert-butyl 2-(2-hydroxyethoxy)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate

3-(7-bromo-8-fluoro-2-(2-hydroxyethoxy)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxy tert-Butyl ester (220.0mg, 0.44mmol) and ((2-fluoro-6-(methoxymethoxy)-8-(4,4,5,5-tetramethyl-1,3,2- Dioxaborolan-2-yl)naphthalen-1-yl)ethynyl)triisopropylsilane (294.7 mg, 0.57 mmol) was dissolved in toluene (6 mL) and water (0.6 mL), and cesium carbonate ( 436.7mg, 1.34mmol) and [n-butylbis(1-adamantyl)phosphine](2-amino-1,1'-biphenyl-2-yl) palladium(II) methanesulfonate (32.9mg, 0.05 mmol), replaced with nitrogen for 3 times, heated to 100° C. and reacted with microwave stirring for 3.5 hours. The reaction solution was directly concentrated. The residue was purified by silica gel column chromatography (eluent: ethyl acetate/petroleum ether=1/1, volume ratio) to obtain the title compound (270.0 mg, yield: 75.9%).

MS m/z (ESI): 803.0 [M+H] ⁺ .

The sixth step: 3-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalene-1-yl) Preparation of tert-butyl 2-(2-oxoethoxy)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate

3-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)-2- (2-Hydroxyethoxy)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester (150.0mg, 0.19mmol) was dissolved in di Dess-Martin oxidant (202.1 mg, 0.48 mmol) was added to methyl chloride (5 mL), and stirred at room temperature for 12 hours. Add saturated aqueous sodium thiosulfate (1 mL) and saturated aqueous sodium bicarbonate (2 mL) to the reaction solution, extract twice with ethyl acetate (20 mL), wash the organic phase with saturated brine (20 mL), wash with anhydrous The organic phase was dried over sodium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: ethyl acetate/petroleum ether=1/1, volume ratio) to obtain the title compound (130.0 mg, yield: 86.9%).

MS m/z (ESI): 801.0 [M+H] ⁺ .

Intermediate Preparation Example 15: Preparation of 2-(piperidin-4-ylmethoxy)ethyl acetate

The first step: the preparation of tert-butyl 4-((2-ethoxy-2-oxoethoxy)methyl)piperidine-1-carboxylate

4-(Hydroxymethyl)piperidine-1-carboxylic acid tert-butyl ester (4.0g, 18.6mmol) was dissolved in dichloromethane (40mL), after adding dipolyrhodium acetate (419.2mg, 0.9mmol), in Ethyl diazoacetate (3.2 g, 28.1 mmol) was added dropwise to the reaction solution at 0° C., replaced with nitrogen three times, then raised to room temperature, and reacted for 16 hours. The reaction solution was poured into water (40 mL), extracted twice with dichloromethane (20 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: ethyl acetate/petroleum ether=1/9, volume ratio) to obtain the title compound (2.6 g, yield: 46.4%).

MS m/z (ESI): 302.0 [M+H] ⁺ .

The second step: the preparation of 2-(piperidin-4-ylmethoxy) ethyl acetate

Add tert-butyl 4-((2-ethoxy-2-oxoethoxy)methyl)piperidine-1-carboxylate (2.6g, 8.6mmol) into 4M ethyl acetate hydrochloric acid solution (30mL) , reacted at 20°C for 2 hours. The reaction solution was directly concentrated under reduced pressure to obtain the hydrochloride salt of the title compound (1.9 g, yield: 95.0%).

MS m/z (ESI): 202.0 [M+H] ⁺ .

Synthetic Example:

Example 1: (2S,4R)-1-((S)-2-(3-(3-((S)-2-(((4-(3,8-diazabicyclo[3.2.1 ]octane-3-yl)-7-(8-chloronaphthalene-1- Base)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)oxy)methyl)pyrrolidin-1-yl)propoxy)propionylamino)- 3,3-Dimethicone Preparation of (butyryl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide (compound 1)

The first step: 3-(7-(8-chloronaphthalen-1-yl)-2-(((S)-1-(3-(3-ethoxy-3-oxopropoxy)propyl )pyrrolidin-2-yl)methoxy)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2. 1] Preparation of benzyl octane-8-carboxylate

3-(2-(((S)-1-(3-(3-ethoxy-3-oxopropoxy)propyl)pyrrolidin-2-yl)methoxy)-5,6 ,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid benzyl ester (329mg, 0.52mmol ) and 1-bromo-8-chloronaphthalene (188 mg, 0.78 mmol) were dissolved in dry 1,4-dioxane (5 mL), and cesium carbonate (674 mg, 2.08 mmol) was added. Under nitrogen protection, tris(dibenzylideneacetone)dipalladium (73 mg, 0.078 mmol) and 2-dicyclohexylphosphine-2',6'-diisopropoxy-1,1'-biphenyl ( 49.2mg, 0.104mmol), and the reaction solution was stirred at 95°C for 10 hours. Water (20 mL) was added to the system, and extracted three times with ethyl acetate (20 mL). The organic phase was dried, concentrated, and purified by silica gel column chromatography (eluent: dichloromethane/methanol=10/1, volume ratio) to obtain the title compound (250 mg, yield: 60.7%).

MS m/z (ESI): 797.4 [M+H] ⁺ .

The second step: 3-(3-((S)-2-(((4-(8-((benzyloxy)carbonyl)-3,8-diazabicyclo[3.2.1]octane-3 -yl)-7-(8-chloronaphthalen-1-yl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)oxy)methyl)pyrrolidine Preparation of -1-yl)propoxyl)propionic acid

3-(7-(8-chloronaphthalen-1-yl)-2-(((S)-1-(3-(3-ethoxy-3-oxopropoxy)propyl)pyrrolidine -2-yl)methoxy)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane Benzyl alkane-8-carboxylate (223mg, 0.28mmol) was dissolved in methanol (4mL) and water (1mL), sodium hydroxide (45mg, 1.12mmol) was added, and the reaction solution was stirred at 30°C for 4 hours. The reaction solution was concentrated, adjusted to pH<5 by adding 1M dilute hydrochloric acid, and extracted three times with ethyl acetate (10 mL). The organic phase was dried and concentrated to give the title compound (210 mg, yield: 97.6%)

MS m/z (ESI): 769.1 [M+H] ⁺ .

The third step: 3-(7-(8-chloronaphthalen-1-yl)-2-(((S)-1-(3-(3-(((S)-1-((2S,4R) -4-Hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutane Alk-2-yl)amino)-3-oxopropoxy)propyl)pyrrolidin-2-yl)methoxy)-5,6,7,8-tetrahydropyrido[3,4-d Preparation of ]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate benzyl ester

3-(3-((S)-2-(((4-(8-((benzyloxy)carbonyl)-3,8-diazabicyclo[3.2.1]octane-3-yl) -7-(8-Chloronaphthalen-1-yl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)oxy)methyl)pyrrolidin-1- (2S,4R)-1-((S)-2-amino-3,3-dimethylbutyryl) -4-Hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide (73mg, 0.16mmol), 2-(7-azobenzotriazole )-N,N,N',N'-tetramethylurea hexafluorophosphate (64.6mg, 0.17mmol) and triethylamine (66mg, 0.65mmol), and stirred at room temperature for 1 hour. The reaction solution was dropped into water (12 mL), and filtered. The filter cake was dissolved with ethyl acetate (10 mL), then dried, filtered and concentrated to give the title compound (128 mg, yield: 83.3%).

MS m/z (ESI): 1181.2 [M+H] ⁺ .

The fourth step: (2S,4R)-1-((S)-2-(3-(3-((S)-2-(((4-(3,8-diazabicyclo[3.2.1 ]octane-3-yl)-7-(8-chloronaphthalen-1-yl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)oxy) Methyl)pyrrolidin-1-yl)propoxy)propionylamino)-3,3-dimethylbutyryl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl) Preparation of benzyl)pyrrolidine-2-carboxamide

3-(7-(8-chloronaphthalen-1-yl)-2-(((S)-1-(3-(3-(((S)-1-((2S,4R)-4- Hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutane-2 -yl)amino)-3-oxopropoxy)propyl)pyrrolidin-2-yl)methoxy)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidine- 4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate benzyl ester (100 mg, 0.085 mmol) was dissolved in dichloromethane (4 mL) and acetonitrile (1 mL), and tris Methyliodosilane (84mg, 0.43mmol), and stirred at room temperature for 4 hours. The reaction solution was concentrated and purified by preparative high performance liquid chromatography to obtain the title compound (15 mg, yield: 16.9%).

MS m/z (ESI): 1047.5 [M+H] ⁺ .

¹ H-NMR(400MHz,DMSO-d ₆ ):δ9.04(s,1H),8.65(s,1H),8.28(s,1H),7.98-7.95(m,2H),7.79(d,J =8Hz,1H),7.64-7.57(m,2H),7.52-7.43(m,5H),7.38(d,J=8Hz,1H),4.61(d,J=12 Hz,1H),4.59-4.46 (m,2H),4.41(s,1H),4.30-4.11(m,4H),4.09(d,J＝8Hz,1H),3.97-3.92(m,2H),3.81-3.51(m,12H) ,3.15-3.04(m,4H),2.91(d,J＝12Hz,1H),2.77-2.73(m,2H),2.51(s,3H),2.40-2.31(m,3H),2.22-2.07( m, 1H), 1.99-1.90 (m, 3H), 1.80-1.64 (m, 8H), 0.96 (s, 9H).

Example 2: (2S,4R)-1-((S)-2-(3-(3-((S)-2-(((4-(3,8-diazabicyclo[3.2.1 ]octane-3-yl)-7-(3-hydroxynaphthalene -1-yl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)oxy)methyl)pyrrolidin-1-yl)propoxy)propionyl Amino)-3,3-di Preparation of methylbutyryl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide (Compound 2)

Using the synthetic route in Example 1, the raw material 1-bromo-8-chloronaphthalene in the first step was replaced with 3-benzyloxy-1-bromonaphthalene to obtain the target product (25 mg, yield: 42.5%).

MS m/z (ESI): 1029.4 [M+H] ⁺ .

¹ H-NMR (400MHz, DMSO-d ₆ ): δ8.95(s, 1H), 8.60(t, J=8Hz, 1H), 8.27(s, 1H), 8.01-7.92(m, 2H), 7.67 (d,J=8Hz,1H),7.44-7.37(m,5H),7.27(t,J=8.4Hz,1H),6.87(s,1H),6.77(s,1H),4.56(d,J ＝8Hz,1H),4.45-4.41(m,2H),4.35(s,1H),4.25-4.20(m,2H),4.06(s,1H),3.96-3.91(m,3H),3.69-3.64 (m,5H),3.60-3.49(m,4H),3.45-3.35(m,3H),2.89-2.84(m,3H),2.77-2.73(m,2H),2.44(s,3H),2.36 -2.21(m,3H),2.22-2.13(m,1H),2.05-1.97(m,2H),1.93-1.78(m,7H),1.69-1.60(m,6H),0.92(s,9H) .

Example 3: (2S,4R)-1-((S)-2-(3-(3-((S)-2-(((4-(3,8-diazabicyclo[3.2.1 ]octane-3-yl)-8-fluoro-7-(3-hydroxyl Naphthalene-1-yl)pyrido[4,3-d]pyrimidin-2-yl)oxy)methyl)pyrrolidin-1-yl)propoxy)propionamide)-3,3-dimethylbutyl Acyl)-4- Preparation of Hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide (Compound 3)

The first step: 3-(7-(3-(benzyloxy)naphthalene-1-yl)-2-(((S)-1-(3-(3-(tert-butoxy)-3-oxo Propoxy)propyl)pyrrolidin-2-yl)methoxy)-8-fluoropyrido[4,3-d]pyrimidin-4-yl)-3,8- diazabicyclo [3.2 .1] Preparation of benzyl octane-8-carboxylate

3-(2-(((S)-1-(3-(3-(tert-butoxy)-3-oxopropoxy)propyl)pyrrolidin-2-yl)methoxy) -7-Chloro-8-fluoropyrido[4,3-d]pyrimidin-4-yl)-3,8- diazabicyclo [3.2.1]octane-8-carboxylic acid benzyl ester (150mg, 199.79μmol), 2-(3-benzyloxynaphthalen-1-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborinane (113.65mg, 299.69μmol), Cesium carbonate (130.19 mg, 399.59 μmol) and Pd(dppf)Cl ₂ (14.62 mg, 19.98 μmol) were added to 1,4-dioxane (5 mL) and water (1 mL), and the reaction system was heated at 100 ° C Stir overnight. The system was cooled to room temperature, and purified by silica gel column chromatography (eluent: ethyl acetate) to obtain the title compound (136 mg, yield: 74.71%).

MS m/z (ESI): 911.4 [M+H] ⁺ .

The second step: 3-(3-((S)-2-(((4-(8-((benzyloxy)carbonyl)-3,8- diazabicyclo [3.2.1]octane- 3-yl)-7-(3-(benzyloxy)naphthalen-1-yl)-8-fluoropyrido[4,3-d]pyrimidin-2-yl)oxy)methyl)pyrrolidin-1 Preparation of -yl)propoxy)propionic acid

3-(7-(3-(benzyloxy)naphthalen-1-yl)-2-(((S)-1-(3-(3-(tert-butoxy)-3-oxopropoxy Base) propyl) pyrrolidin-2-yl) methoxy) -8-fluoropyrido [4,3-d] pyrimidin-4-yl) -3,8- diazabicyclo [3.2.1] Benzyl octane-8-carboxylate (136 mg, 149.28 μmol) was dissolved in dichloromethane (5 mL). The temperature of the reaction system was controlled at about 0°C in an ice bath, and trifluoroacetic acid (1 mL) was added dropwise. After dropping, stirred at 25°C for 2 hours, concentrated under reduced pressure and spin-dried to obtain the title compound (151 mg, yield: 93.40%), which was directly used in the next step.

MS m/z (ESI): 855.4 [M+H] ⁺ .

The third step: 3-(7-(3-(benzyloxy)naphthalene-1-yl)-8-fluoro-2-(((S)-1-(3-(3-(((S)- 1-((2S,4R)-4-Hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-3,3-di Methyl-1-oxobutan-2-yl)amino)-3-oxopropoxy)propyl)pyrrolidin-2-yl)methoxy)pyrido[4,3-d]pyrimidine- Preparation of 4-yl)-3,8- diazabicyclo [3.2.1]octane-8-carboxylate benzyl ester

3-(3-((S)-2-(((4-(8-((benzyloxy)carbonyl)-3,8- diazabicyclo [3.2.1]octane-3- Base) -7-(3-(benzyloxy)naphthalen-1-yl)-8-fluoropyrido[4,3-d]pyrimidin-2-yl)oxy)methyl)pyrrolidin-1-yl )propoxy)propionic acid (118mg, 108.96μmol) and (2S,4R)-1-((S)-2-amino-3,3-dimethylbutyryl)-4-hydroxyl-N-(4 -(4-Methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide (56.29 mg, 130.75 μmol) was sequentially added to DMF (2 mL). The temperature of the reaction system was controlled at about 0°C in an ice bath, and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (53.86 mg, 141.64μmol) and N,N-diisopropylethylamine (71.82mg, 555.67μmol). After the addition was complete, the reaction system was stirred at this temperature for 1 hour, and added dropwise into ice water (10 mL). The precipitated solid was filtered with suction, and the filter cake was dried to obtain the title compound (108 mg, yield: 78.20%).

MS m/z (ESI): 1267.6 [M+H] ⁺ .

The fourth step: (2S,4R)-1-((S)-2-(3-(3-((S)-2-(((4-(3,8- diazabicyclo [3.2. 1] Octane-3-yl)-8-fluoro-7-(3-hydroxynaphthalen-1-yl)pyrido[4,3-d]pyrimidin-2-yl)oxy)methyl)pyrrolidine- 1-yl)propoxy)propionamide)-3,3-dimethylbutyryl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2 - Preparation of formamide

3-(7-(3-(Benzyloxy)naphthalen-1-yl)-8-fluoro-2-(((S)-1-(3-(3-(((S)-1-( (2S,4R)-4-Hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl- 1-oxobutan-2-yl)amino)-3-oxopropoxy)propyl)pyrrolidin-2-yl)methoxy)pyridyl[4,3-d]pyrimidine-4- Base)-3,8- diazabicyclo [3.2.1]octane-8-carboxylate benzyl ester (60mg, 47.34μmol) was added into a mixed solvent of dichloromethane (2mL) and acetonitrile (6mL), in Iodotrimethylsilane (37.89 mg, 189.35 μmol) was added dropwise under ice bath. After dropping, the mixture was stirred at 10°C for 6 hours. The reaction system was quenched with ice water, concentrated at low temperature, and purified by preparative high performance liquid chromatography to obtain the title compound (4.93 mg, yield: 9.08%).

MS m/z (ESI): 1043.5 [M+H] ⁺ .

¹ H-NMR (400MHz, CD ₃ OD): δ9.15(s, 1H), 8.86(s, 1H), 8.41(br, 1H), 7.76(d, J=8.0Hz, 1H), 7.53(d ,J＝8.0Hz,1H),7.44-7.36(m,5H),7.29-7.21(m,3H),5.58(br,1H),4.79-4.72(m,3H),4.61-4.29(m,5H ),4.00(s,2H),3.86-3.81(m,3H),3.76-3.56(m,5H),3.48-3.41(m,2H),3.16-3.07(m,4H),2.63-2.49(m ,2H), 2.44(s,3H), 2.22-1.98(m,13H), 0.98(s,9H).

Example 4: (2S,4R)-1-((S)-2-(5-((S)-2-(((4-(3,8-diazabicyclo[3.2.1]octane -3-yl)-7-(3-hydroxynaphthalene-1- Base)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)oxy)methylpyrrolidin-1-yl)pentanoylamino)-3,3-di Methylbutyryl)- Preparation of 4-Hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide (Compound 4)

The first step: 4-((8-(benzyloxy)carbonyl)-3,8-diazabicyclo[3.2.1]octane-3-yl))-2-(S)-1-( 5-ethoxy-5-oxopentyl)pyrrolidin-2-yl)methoxy)-5,6-dihydropyrido[3,4-d]pyrimidine-7(8H)-carboxylic acid tert Preparation of butyl ester

4-(8-((Benzyloxy)carbonyl)-3,8-diazabicyclo[3.2.1]octyl-3-yl)-2-chloro-5,6-dihydropyrido[3 ,4-d]pyrimidine-7(8H)-carboxylate tert-butyl ester (1.26g, 2.33mmol) and (S)-ethyl 5-(2-(hydroxymethyl)pyrrolidin-1-yl)pentanoate (1.34g, 4.67mmol) was dissolved in 1,4-dioxane (20mL), cesium carbonate (2.28g, 7.00mmol) was added, replaced with nitrogen three times, Pd(dppf)Cl ₂ (340mg, 0.466mmol) was added ) and 4,5-bisdiphenylphosphine-9,9-dimethylxanthene (405mg, 0.7mmol), stirred at 100°C for 12 hours. After the reaction system was cooled to room temperature, it was extracted three times with ethyl acetate (30 mL), concentrated to remove the solvent, and the obtained residue was purified by silica gel column chromatography (eluent: dichloromethane/methanol=10/1, volume ratio), The title compound (2.0 g, yield: 72.0%) was obtained.

MS m/z (ESI): 707.4 [M+H] ⁺ .

The second step: 3-(2-(S)-1-(5-ethoxy-5-oxopentyl)pyrrolidin-2-yl)methoxy)-5,6,7,8-tetra Preparation of Benzyl Hydropyrido[3,4-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate

4-((8-(benzyloxy)carbonyl)-3,8-diazabicyclo[3.2.1]octane-3-yl))-2-(S)-1-(5-ethane Oxy-5-oxopentyl)pyrrolidin-2-yl)methoxy)-5,6-dihydropyrido[3,4-d]pyrimidine-7(8H)-carboxylic acid tert-butyl ester ( 1.0g, 1.42mmol) was dissolved in trifluoroacetic acid (10mL) and dichloromethane (20mL), and stirred at 20°C for 2 hours. Direct concentration gave the title compound (0.6 g, yield: 69.9%).

MS m/z (ESI): 607.4 [M+H] ⁺ .

The third step: 3-(7-(3-(benzyloxy)naphthalen-1-yl)-2-(((S)-1-(5-ethoxy-5-oxopentyl)pyrrolidine -2-yl)methoxy)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane Preparation of benzyl alkane-8-carboxylate

3-(2-(S)-1-(5-ethoxy-5-oxopentyl)pyrrolidin-2-yl)methoxy)-5,6,7,8-tetrahydropyrido [3,4-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate benzyl ester (200.0mg, 0.33mmol) and 3-benzyloxy- 1-bromonaphthalene (155mg, 0.5mmol) was dissolved in toluene (10mL), cesium carbonate (215mg, 0.66mmol) was added, replaced with nitrogen three times, methanesulfonic acid (2-dicyclohexylphosphino-2',4 ',6'-tri-isopropyl-1,1'-biphenyl)(2'-amino-1,1'-biphenyl-2-yl)palladium(II) (28mg, 0.033mmol), in Stir at 100°C for 8 hours. After the reaction system was cooled to room temperature, it was extracted three times with ethyl acetate (10 mL), concentrated to remove the solvent, and the obtained residue was purified by silica gel column chromatography (eluent: dichloromethane/methanol=10/1, volume ratio), The title compound (70.0 mg, yield: 25.6%) was obtained.

MS m/z (ESI): 839.4 [M+H] ⁺ .

The fourth step: 5-((S)-2-(((4-(8-((benzyloxy)carbonyl)-3,8-diazabicyclo[3.2.1]octane-3-yl) -7-(3-(Benzyloxy)naphthalen-1-yl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)oxy)methyl)pyrrole Preparation of alk-1-yl)pentanoic acid

3-(7-(3-(Benzyloxy)naphthalen-1-yl)-2-(((S)-1-(5-ethoxy-5-oxopentyl)pyrrolidin-2- Base)methoxy)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8 - Benzyl carboxylate (70mg, 0.083mmol) was dissolved in ethanol (5mL), and an aqueous solution of sodium hydroxide (16.6mg, 0.415mmol) was added dropwise, and reacted at 25°C for 4 hours. The organic solvent was concentrated, adjusted to pH=5 with hydrochloric acid, extracted three times with ethyl acetate (10 mL), and concentrated to obtain the title compound (60.9 mg, yield: 90.0%).

MS m/z (ESI): 811.4 [M+H] ⁺ .

The fifth step: 3-(7-(3-(benzyloxy)naphthalen-1-yl)-2-(S)-1-(5-(S)-1-(2S,4R)-4- Hydroxy-2-(4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutane-2- Base)amino)-5-oxopentyl)pyrrolidin-2-yl)methoxy)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)- Preparation of benzyl 3,8-diazabicyclo[3.2.1]octane-8-carboxylate

5-((S)-2-(((4-(8-((benzyloxy)carbonyl)-3,8-diazabicyclo[3.2.1]oct-3-yl)-7-( 3-(Benzyloxy)naphthalen-1-yl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)oxy)methyl)pyrrolidin-1- base) valeric acid (60.9mg, 75μmol) was dissolved in N,N-dimethylformamide (3mL), and (2S,4R)-1-(S)-2-amino-3,3-dimethyl Butyryl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide (38.5mg, 82.5μmol), 2-(7-azobenzene Triazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (37.05mg, 97.5μmol) and triethylamine (37.9mg, 375μmol), stirred at 25°C for 1 hour. The reaction solution was dropped into water, and a solid was precipitated, filtered, and the filter cake was dried to obtain the title compound (35.0 mg, yield: 38.1%).

MS m/z (ESI): 1223.5 [M+H] ⁺ .

Step 6: (2S,4R)-1-((S)-2-(5-((S)-2-(((4-(3,8-diazabicyclo[3.2.1]octane -3-yl)-7-(3-hydroxynaphthalen-1-yl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)oxy)methylpyrrole Alkyl-1-yl)pentanoylamino)-3,3-dimethylbutyryl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidin-2- Preparation of formamide.

3-(7-(3-(Benzyloxy)naphthalen-1-yl)-2-(S)-1-(5-(S)-1-(2S,4R)-4-hydroxyl-2 -(4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino )-5-oxopentyl)pyrrolidin-2-yl)methoxy)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)-3,8 - Benzyl diazabicyclo[3.2.1]octane-8-carboxylate (30mg, 24.5μmol) was dissolved in dichloromethane (1mL), cooled to 0°C, hydrobromic acetic acid solution (19.33 mg, 71.67μmol), stirred at 0°C for 20 minutes, discarded the supernatant, diluted with methanol, adjusted pH=7-8 with sodium bicarbonate solution and concentrated the solution, the residue was purified by preparative high performance liquid chromatography , to obtain the title compound (12 mg, yield: 48.8%).

MS m/z (ESI): 999.5 [M+H] ⁺ .

¹ H-NMR (400MHz, CD ₃ OD) δ8.84(s, 1H), 8.54(s, 1H), 8.04(d, J=8.0Hz, 1H), 7.61(d, J=8.0Hz, 1H) ,7.44-7.42(m,2H),7.39-7.34(m,3H),7.27-7.23(m,2H),6.85-6.79(m,2H),4.63-4.47(m,6H),4.39-4.29( m,4H),4.13-4.05(m,4H),3.91-3.88(m,2H),3.80-3.72(m,4H),3.52-3.43(m,2H),3.15-3.10(m,2H), 2.93(s,2H),2.44(s,3H),2.36-2.29(m,2H),2.21-2.17(m,2H),2.10-2.07(m,2H),2.04-1.93(m,6H), 1.69-1.62(m,4H),1.01(s,9H).

Example 5: (2S,4R)-1-((S)-2-(6-((S)-2-(((4-(3,8-diazabicyclo[3,2.1]octane -3-yl)-7-(3-hydroxynaphthalene-1- Base)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)oxy)methyl)pyrrolidin-1-yl)hexanoylamino)-3,3- Dimethylbutyryl)- Preparation of 4-Hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide (compound 5)

Using the synthetic route in Example 4, the first step reaction raw material (S)-5-(2-(hydroxymethyl)pyrrolidin-1-yl)ethyl valerate was replaced by (S)-6-(2 -Ethyl (hydroxymethyl)pyrrolidin-1-yl)hexanoate to obtain the title compound (20 mg, yield: 46.1%).

MS m/z (ESI): 1013.5 [M+H] ⁺ .

¹ H-NMR (400MHz, CD ₃ OD) δ8.86(s, 1H), 8.53(s, 1H), 8.05(d, J=8.0Hz, 1H), 7.62(d, J=8.0Hz, 1H) ,7.45-7.34(m,6H),7.27-7.24(m,1H),6.86(d,J=2.0Hz,1H),6.79(d,J=2.0Hz,1H),4.60-4.30(m,9H ),4.14-4.10(m,4H),3.89-3.75(m,5H),2.94-2.67(m,6H),2.45(s,3H),2.36-2.12(m,5H),2.09-1.89(m ,8H), 1.84-1.81(m,2H), 1.66-1.61(m,4H), 1.41-1.35(m,2H), 1.00(s,9H).

Example 6: (2S,4R)-1-((S)-2-(7-((S)-2-(((4-(3,8-diazabicyclo[3.2.1]octane -3-yl)-7-(3-hydroxynaphthalene-1- Base)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)oxy)methyl)pyrrolidin-1-yl)heptanoylamino)-3,3- Dimethylbutyryl)- Preparation of 4-Hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide (Compound 6)

Using the synthetic route of Example 4, the first step reaction raw material (S)-5-(2-(hydroxymethyl)pyrrolidin-1-yl) ethyl valerate was replaced by (S)-7-(2- (Hydroxymethyl)pyrrolidin-1-yl)ethyl heptanoate to obtain the title compound (24 mg, yield: 49.4%).

MS m/z (ESI): 1027.5 [M+H] ⁺ .

¹ H-NMR (400MHz, CD ₃ OD) δ8.85(s, 1H), 8.54(s, 1H), 8.05(d, J=8.0Hz, 1H), 7.62(d, J=8.0Hz, 1H) ,7.45-7.23(m,6H),6.86-6.78(m,2H),4.61-4.31(m,9H),4.13-4.10(m,4H),3.90-3.77(m,5H),3.61-3.31( m,5H),3.20-2.60(m,6H),2.45(s,3H),2.33-1.76(m,12H),1.67-1.55(m,4H),1.45-1.31(m,4H),1.01( s, 9H).

Example 7: (2S,4R)-1-((S)-2-(2-((1-(2-((4-(3,8-diazabicyclo[3.2.1]octane- 3-yl)-7-(8-ethynyl-7-fluoro -3-Hydroxynaphthalen-1-yl)-8-fluoropyrido[4,3-d]pyrimidin-2-yl)oxy)ethyl)piperidin-4-yl)methoxy)acetamide-3 ,3-Dimethylbutane Preparation of acyl-4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (compound 7)

The first step: 3-(2-(2-(4-((2-ethoxy-2-oxoethoxy)methylpiperidin-1-yl)ethoxy)-8-fluoro-7 -(7-fluoro-3-(methoxymethoxy)-8-(triisopropylsilyl)ethynyl)naphthalen-1-yl)pyrido[4,3-d]pyrimidine-4- Preparation of tert-butyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate

To a 50mL round bottom flask, add 3-(7-chloro-2-(2-(4-((2-ethoxy-2-oxoethoxy)methyl)piperidin-1-yl )ethoxy)-8-fluoropyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester (146mg ,0.23mmol), 2-[2-fluoro-6-(methoxymethoxy)-8-(4,4,5,5-tetramethyl-1,3,2-dioxaborol Alk-2-yl)-1-naphthyl]ethynyltriisopropylsilane (180 mg, 0.35 mmol), potassium phosphate (98 mg, 0.46 mmol), 1,4-dioxane (5 mL) and water (0.5 mL), replaced with nitrogen three times, to which was added [(bis(1-adamantyl)butylphosphino)-2-(2′-amino-1,1′-biphenyl)]palladium ( II) (36mg, 0.05mmol), replaced with nitrogen three times, heated the reaction mixture to 100°C, and reacted for 12 hours under stirring. Cool to room temperature, add water thereto, extract three times with ethyl acetate (10 mL), wash the organic phase three times with saturated brine (10 mL), dry the obtained organic phase over anhydrous sodium sulfate, filter, and spin dry to obtain the title compound ( 185 mg, yield: 80.2%).

MS m/z (ESI): 987.5 [M+H] ⁺ .

The second step: 2-((1-(2-((4-(8-(tert-butoxycarbonyl)-3,8-diazabicyclo[3.2.1]octane-3-yl)-8- Fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalene-1-yl)pyrido[4,3-d] Preparation of pyrimidin-2-yl)oxy)ethyl)piperidin-4-yl)methoxy)acetic acid

To a 50 mL round bottom flask, add 3-(2-(2-(4-((2-ethoxy-2-oxoethoxy)methylpiperidin-1-yl)ethoxy) -8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-(triisopropylsilyl)ethynyl)naphthalen-1-yl)pyrido[4,3- d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate tert-butyl ester (160 mg, 0.16 mmol), methanol (2 mL), water (1 mL) and hydrogen Lithium oxide (34mg, 0.8mmol), the reaction mixture was stirred at 25°C for 4 hours. The solvent was evaporated under reduced pressure, water (2 mL) was added thereto, adjusted to pH=5-6 with 1N dilute hydrochloric acid, left to stand, and suction filtered to obtain the title compound (138 mg, yield: 90%).

MS m/z (ESI): 959.5 [M+H] ⁺ .

The third step: 3-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalene-1-yl) -2-(2-(4-((2-(((S)-1-((2S,4R)-4-hydroxy-2-(((S)-1-(4-(4-methyl Thiazol-5-yl)phenyl)ethyl)))carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-2- Oxoethoxy)methyl)piperidin-1-yl)ethoxy)pyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]- Preparation of tert-butyl octane-8-carboxylate

To a 50mL round bottom flask, add 2-(1-(2-((4-(8-(tert-butoxycarbonyl)-3,8-diazabicyclo[3.2.1]octane-3- Base)-8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)pyrido[4 ,3-d]pyrimidin-2-yl)oxy)ethyl)piperidin-4-yl)methoxy)acetic acid (134mg, 0.14mmol), (2S,4R)-1-[(2S)-2 -Amino-3,3-dimethylbutyryl]-4-hydroxy-N-[(1S)-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]ethyl base]pyrrolidine-2-carboxamide (68mg, 0.14mmol), N,N-diisopropylethylamine (36mg, 0.28mmol) and N,N-dimethylformamide (2mL), the reaction mixture was cooled To 0°C, 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (80mg, 0.21mmol) was added in batches, and The reaction mixture was warmed up to 25°C and reacted for 12 hours with stirring. The reaction mixture was poured into water, extracted three times with ethyl acetate (10 mL), the organic phase was washed twice with saturated brine (10 mL), the obtained organic phase was dried over anhydrous sodium sulfate, filtered, and spin-dried to obtain the title compound (166 mg, yield: 85.7%).

MS m/z (ESI): 1386.7 [M+H] ⁺ .

The fourth step: 3-(7-(8-ethynyl-7-fluoro-3-(methoxymethoxy)naphthalene-1-yl)-8-fluoro-2-(2-(4-(( 2))-(((S)-1-((2S,4R)-4-hydroxy-2-(((S)-1-(4-(4-methylthiazol-5-yl)phenyl) Ethyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-2-oxoethoxy)methyl)piperidine Preparation of -1-yl)ethoxy)pyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester

Into a 50 mL round bottom flask, add 3-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl) Naphthalene-1-yl)-2-(2-(4-((2-(((S)-1-((2S,4R)-4-hydroxy-2-(((S)-1-(4 -(4-methylthiazol-5-yl)phenyl)ethyl)))carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl )amino)-2-oxoethoxy)methyl)piperidin-1-yl)ethoxy)pyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo [3.2.1]-Octane-8-carboxylic acid tert-butyl ester (166mg, 0.12mmol) and dichloromethane (2mL), add 1M tetrabutylammonium fluoride tetrahydrofuran solution (0.24mL, 0.24mL) to the reaction solution mmol), the reaction mixture was stirred at 25°C for 12 hours. The solvent was evaporated under reduced pressure, the reaction mixture was poured into water, extracted three times with ethyl acetate (10 mL), the organic phase was washed twice with saturated brine (10 mL), the obtained organic phase was dried over anhydrous sodium sulfate, filtered, Spin-dried to obtain the title compound (142 mg, yield: 96%).

MS m/z (ESI): 1229.6 [M+H] ⁺ .

The fifth step: (2S,4R)-1-((S)-2-(2-((1-(2-((4-(3,8-diazabicyclo[3.2.1]octane- 3-yl)-7-(8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluoropyrido[4,3-d]pyrimidin-2-yl)oxy)ethyl )piperidin-4-yl)methoxy)acetamide-3,3-dimethylbutyryl-4-hydroxyl-N-((S)-1-(4-(4-methylthiazole-5- base) phenyl) ethyl) pyrrolidine-2-carboxamide preparation

Into a 50mL round bottom flask, add 3-(7-(8-ethynyl-7-fluoro-3-(methoxymethoxy)naphthalene-1-yl)-8-fluoro-2-(2 -(4-((2))-(((S)-1-((2S,4R)-4-Hydroxy-2-(((S)-1-(4-(4-methylthiazole-5 -yl)phenyl)ethyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-2-oxoethoxy )methyl)piperidin-1-yl)ethoxy)pyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxy Acetyl tert-butyl ester (135mg, 0.11mmol) and 1,4-dioxane (2mL), to which was added 4M hydrogen chloride in 1,4-dioxane solution (0.2mL, 0.8mmol), and the reaction mixture was The reaction was stirred at 25°C for 12 hours. The solvent was evaporated under reduced pressure, water was added thereto, adjusted to pH = 8-9 with saturated aqueous sodium bicarbonate solution, allowed to stand, and filtered with suction to obtain a solid, which was purified by preparative high-performance liquid chromatography to obtain the title compound (42 mg, Yield: 35%).

MS m/z (ESI): 1085.5 [M+H] ⁺ .

¹ H-NMR (400MHz,DMSO-d ₆ )δ10.16(s,1H),9.03(s,1H),8.99(s,1H),8.47(d,J=7.6Hz,1H),7.97(dd ,J=9.2,6.0Hz,1H),7.48-7.27(m,7H),7.17(d,J=2.4Hz,1H),5.15(d,J=3.6Hz,1H),4.92-4.85(m, 1H), 4.45-4.27(m, 7H), 3.95-3.85(m, 3H), 3.64-3.54(m, 6H), 3.32(s, 3H), 2.96(d, J＝10.8Hz, 2H), 2.70 -2.51(m,3H),2.45(s,3H),2.09-1.99(m,3H),1.78-1.45(m,9H),1.36-1.19(m,5H),0.93(s,9H).

Example 8: (2S,4R)-1-(2-(3-((1-(2-(4-(3,8-diazabicyclo[3.2.1]octan-3-yl)- 7-(8-Ethynyl-7-fluoro-3-hydroxy Basenaphth-1-yl)-8-fluoropyrido[4,3-d]pyrimidin-2-yl)oxy)ethyl)piperidin-4-yl)methoxy)isoxazol-5-yl )-3-methylbutyryl Preparation of -4-hydroxyl-N-(S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (compound 8)

The first step: 3-(7-chloro-8-fluoro-2-(2-hydroxyethoxy)pyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[ 3.2.1] Preparation of tert-butyl octane-8-carboxylate

3-(2,7-dichloro-8-fluoropyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid Tert-butyl ester (200mg, 457.64μmol) was dissolved in DMF (1.00mL), cesium carbonate (456.45mg, 1.37mmol) and ethylene glycol (85.21mg, 1.37mmol) were added, and stirred at 25°C for 2 hours. Add water to the reaction solution, extract three times with ethyl acetate (20 mL), wash the organic phase with saturated brine (20 mL), dry the organic phase with anhydrous sodium sulfate, and concentrate under reduced pressure to obtain the title compound (200 mg, yield: 94.3 %).

MS m/z (ESI): 454.2 [M+H] ⁺ .

The second step: 3-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-(triisopropylsilyl)ethynyl)naphthalene-1-yl)- tert-butyl 2-(2-hydroxyethoxy)pyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate preparation

3-(7-chloro-8-fluoro-2-(2-hydroxyethoxy)pyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2. 1] Octane-8-carboxylate tert-butyl ester (190 mg, 418 μmol) was dissolved in 1,4-dioxane (10 mL) and water (1 mL), and 2-[2-fluoro-6-(methoxy methoxy)-8-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-naphthyl]ethynyltriisopropane Silane (276.31mg, 533.71μmol), potassium phosphate (227mg, 1.07mmol) and [n-butylbis(1-adamantyl)phosphine](2-amino-1,1'-biphenyl-2 -yl) palladium(II) (25.9 mg, 35.58 μmol), replaced with nitrogen three times, and stirred at 100° C. for 15 hours. Cool the reaction solution to room temperature, add water (20 mL) to dilute, extract three times with ethyl acetate (20 mL), wash the organic phase with saturated brine (20 mL), dry the organic phase with anhydrous sodium sulfate, and concentrate under reduced pressure, the residue Purification by silica gel column chromatography (eluent: petroleum ether/ethyl acetate=1/1, volume ratio) gave the title compound (200 mg, yield: 59.5%).

MS m/z (ESI): 804.4 [M+H] ⁺ .

The third step: 3-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-(triisopropylsilyl)ethynyl)naphthalene-1-yl)- tert-Butyl 2-(2-oxoethoxy)pyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate preparation of

3-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-(triisopropylsilyl)ethynyl)naphthalen-1-yl)-2-( 2-Hydroxyethoxy)pyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester (50mg, 62.2 μmol) was dissolved in dichloromethane (2 mL), then Dess-Martin oxidant (100 mg, 236.32 μmol) was added and stirred at 25°C for 18 hours. Concentrate the reaction solution, then add ethyl acetate (30 mL) to dilute the residue, wash twice with water (30 mL), collect the organic phase, wash the organic phase with saturated brine (150 mL), dry the organic phase with anhydrous sodium sulfate, and reduce Concentration under reduced pressure gave the title compound (30 mg, yield: 60.1%).

MS m/z (ESI): 802.4 [M+H] ⁺ .

The fourth step: 3-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalene-1-yl) -2-(2-(4-(((5-(1-((2S,4R)-4-hydroxy-2-(((S)-1-(4-methylthiazol-5-yl)benzene Base) ethyl) carbamoyl) pyrrolidin-1-yl) -3-methyl-1-oxobutan-2-yl) isoxazol-3-yl) oxy) methyl) piperidine- Preparation of 1-yl)ethoxy)pyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester

3-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-(triisopropylsilyl)ethynyl)naphthalen-1-yl)-2-( 2-Oxoethoxy)pyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester (22mg, 27.4 μmol) was dissolved in dichloromethane (499.77 μL) and isopropanol (499.77 μL), and (2S,4R)-4-hydroxy-1-(3-methyl-2-(3-(piperidine- 4-ylmethoxy)isoxazol-5-yl)butyryl)-N-(S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine- 2-Formamide (16.34mg, 27.4μmol), sodium acetate (10.69mg, 130.30μmol), after stirring for 15 minutes under ice bath, then added 2-picoline borane (14.08mg, 130.30μmol), at 25 Stir at °C for 15 hours. The reaction solution was concentrated, and the residue was purified by silica gel column chromatography (eluent: dichloromethane/methanol=10/1, volume ratio) to obtain the title compound (20 mg, yield: 52.8%).

MS m/z (ESI): 1381.7 [M+H] ⁺ .

The fifth step: 3-(7-(8-ethynyl-7-fluoro-3-(methoxymethoxy)naphthalene-1-yl)-8-fluoro-2-(4-(((5- (1-((2S,4R)-4-Hydroxy-2-(((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidine -1-yl)-3-methyl-1-oxobutan-2-yl)isoxazol-3-yl)methyl)piperidin-1-yl)ethoxy)pyrido[4,3 Preparation of -d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester

3-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)-2- (2-(4-(((5-(1-((2S,4R)-4-hydroxyl-2-(((S)-1-(4-methylthiazol-5-yl)phenyl)ethyl Base) carbamoyl) pyrrolidin-1-yl)-3-methyl-1-oxobutan-2-yl) isoxazol-3-yl) oxy)methyl)piperidin-1-yl )ethoxy)pyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester (20mg, 13.75μmol) Dissolve in dichloromethane (1 mL), then add tetrabutylammonium fluoride (1M, 27.50 μL), protect with nitrogen, and stir at 25°C for 8 hours. Water (10 mL) and ethyl acetate (10 mL) were added to the reaction solution for extraction twice, the organic phase was washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain the title compound (20 mg, Yield: 94.9%).

MS m/z (ESI): 1225.5 [M+H] ⁺ .

The sixth step: (2S,4R)-1-(2-(3-((1-(2-(4-(3,8-diazabicyclo[3.2.1]octane-3-yl)- 7-(8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluoropyrido[4,3-d]pyrimidin-2-yl)oxy)ethyl)piperidine-4 -yl)methoxy)isoxazol-5-yl)-3-methylbutyryl)-4-hydroxyl-N-(S)-1-(4-(4-methylthiazol-5-yl) Preparation of phenyl)ethyl)pyrrolidine-2-carboxamide

3-(7-(8-ethynyl-7-fluoro-3-(methoxymethoxy)naphthalen-1-yl)-8-fluoro-2-(4-(((5-(1- ((2S,4R)-4-Hydroxy-2-(((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidine-1- Base)-3-methyl-1-oxobutan-2-yl)isoxazol-3-yl)methyl)piperidin-1-yl)ethoxy)pyrido[4,3-d] Pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate tert-butyl ester (23.53 mg, 16.32 μmol) was dissolved in dichloromethane (2 mL) and trifluoroacetic acid was added (310 μL), stirred at 25° C. for 2 hours. Concentrate the reaction solution, then dilute the residue with ethyl acetate (10 mL), adjust the pH to 8-9 with sodium bicarbonate solution, separate the layers, dry the organic phase with anhydrous sodium sulfate, concentrate, and pass the crude product through high performance liquid chromatography Purified by method to obtain the title compound (3.89 mg, yield: 18.7%).

MS m/z (ESI): 1081.4 [M+H] ⁺ .

¹ H NMR (400MHz, CD ₃ OD) δ9.04 (d, J = 8.0Hz, 1H), 8.88-8.83 (m, 1H), 8.46 (s, 1H), 7.85 (dd, J = 16.0, 8.0Hz ,1H),7.45-7.28(m,6H),7.20(d,J=4Hz,1H),5.86(d,J=8Hz,1H),4.99(dd,J=16.0,8.0Hz,2H),4.74 -4.61(m,4H),4.57-4.49(m,1H),4.43(s,1H),3.95(s,2H),3.88-3.75(m,4H),3.69(d,J＝8.0Hz,1H ),3.65-3.59(m,1H),3.37(d,J=8.0Hz,2H),3.13(dd,J=4.0,4.0Hz,2H),2.63-2.32(m,7H),2.22-2.17( m,1H),2.05-1.89(m,5H),1.79(d,J=12.0Hz,3H),1.52-1.45(m,4H),1.31(d,J=16.0Hz,4H),1.06-1.02 (m, 2H), 0.95(t, J=8.0Hz, 2H), 0.90(t, J=8.0Hz, 1H).

Example 9: (2S,4R)-1-(2S)-2-(1-(2-(4-(3,8-diazabicyclo[3.2.1]octane-3-yl)-7 -(8-ethynyl-7-fluoronaphthalene- 1-yl)-8-fluoropyrido[4,3-d]pyrimidin-2-yl)oxy)ethyl)piperidin-4-yl)methoxy)acetamide)-3,3-dimethyl butyryl)-4- Preparation of Hydroxy-N-(S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (Compound 9)

The first step: 3-(2-(4-(2-ethoxy-2-oxoethoxy)methyl)piperidin-1-yl)ethoxy)-8-fluoro-7-(7 -Fluoro-8-(triisopropylsilyl)ethynyl)naphthalene-1-yl)pyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2. 1] Preparation of tert-butyl octane-8-carboxylate

Into a 50mL single-necked bottle, sequentially add tert-butyl 3-(7-chloro-2-(2-(4-(2-ethoxy-2-oxoethoxy)methyl)piperidin-1-yl )ethoxy)-8-fluoropyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester (50mg ,78.48μmol), ((2-fluoro-8-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalene-1-yl) Ethynyl)triisopropylsilane (53.26 mg, 117.71 μmol), potassium phosphate (33.27 mg, 156.95 μmol), 1,4-dioxane (10 mL) and water (2 mL), to which methanesulfonic acid was added [ n-Butylbis(1-adamantyl)phosphine](2-amino-1,1'-biphenyl-2-yl)palladium(II) (11.43mg, 15.70μmol) was replaced three times with nitrogen, and the reaction mixture The reaction was stirred at 100°C for 12 hours. After the reaction solution was cooled to room temperature, water was added to the reaction solution, extracted twice with ethyl acetate (20 mL), the organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The title compound (39.6 mg, yield: 54.43%) was obtained.

MS m/z (ESI): 927.5 [M+H] ⁺ .

The second step: 2-(1-(2-(8-(tert-butoxycarbonyl)-3,8-diazabicyclo[3.2.1]octane-3-yl)-8-fluoro-7-( 7-fluoro-8-(triisopropylsilyl)ethynyl)naphthalen-1-yl)pyrido[4,3-d]pyrimidin-2-yl)oxy)ethyl)piperidin-4- base) the preparation of methoxy) acetic acid

3-(2-(4-(2-ethoxy-2-oxyethoxy)methyl)piperidin-1-yl)ethoxy)-8-fluoro-7-(7-fluoro-8 -(triisopropylsilyl)ethynyl)naphthalen-1-yl)pyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane tert-butyl-8-carboxylate (39.6mg, 42.71μmol) was dissolved in methanol (5mL) and water (1mL), then lithium hydroxide monohydrate (8.97mg, 170.84μmol) was added, and stirred at 25°C for 12 hours. After concentrating the reaction solution, add water to the reaction solution, adjust the pH to 5-6 with 1M HCl solution, then extract three times with ethyl acetate (20 mL), wash the organic phase with saline (30 mL), wash with anhydrous sodium sulfate The organic phase was dried and concentrated under reduced pressure. The residue was purified by flash column chromatography (eluent: dichloromethane/methanol=20/3, volume ratio) to obtain the title compound (18 mg, yield: 46.88%).

MS m/z (ESI): 899.5 [M+H] ⁺ .

The third step: 3-(8-fluoro-7-(7-fluoro-8-(triisopropylsilyl)ethynyl)naphthalene-1-yl)-2-(4-(2-(S) -1-(2S,4R)-4-Hydroxy-2-(S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidine-1- base)-3,3-dimethyl-1-oxobutane-2-amino)-2-oxyl)methyl)piperidin-1-yl)ethoxy)pyrido[4,3-d Preparation of tert-butyl ]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate

2-(1-(2-(8-(tert-butoxycarbonyl)-3,8-diazabicyclo[3.2.1]octyl-3-yl)-8-fluoro-7-(7-fluoro -8-(triisopropylsilyl)ethynyl)naphthalen-1-yl)pyrido[4,3-d]pyrimidin-2-yl)oxy)ethyl)piperidin-4-yl)methyl Oxy)acetic acid (18 mg, 22.02 μmol) was dissolved in N,N-dimethylformamide (4 mL), and (2S,4R)-1-[(2S)-2-amino-3,3-dimethyl Butyryl]-4-hydroxy-N-[(1S)-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide ( 10.59mg, 22.02μmol), triethylamine (10.13mg, 100.09μmol), 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (9.90 mg, 26.02 μmol), replaced with nitrogen three times, and stirred at 25°C for 12 hours. Water was added to the reaction solution, and extracted three times with ethyl acetate (10 mL), the organic phase was washed with brine (10 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The title compound (14.5 mg, yield: 54.63%) was obtained.

MS m/z (ESI): 1325.7 [M+H] ⁺ .

The fourth step: 3-(7-(8-ethynyl-7-fluoronaphthalen-1-yl)-8-fluoro-2-(2-(S)-1-(2S,4R)-4-hydroxyl- 2-(S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1- Oxobutan-2-yl)amino)-2-oxoethoxy)piperidin-1-yl)ethoxy)pyrido[4,3-d]pyrimidin-4-yl)-3,8 -Preparation of tert-butyl diazabicyclo[3.2.1]octane-8-carboxylate

3-(8-fluoro-7-(7-fluoro-8-(triisopropylsilyl)ethynyl)naphthalene-1-yl)-2-(4-(2-(S)-1- (2S,4R)-4-Hydroxy-2-(S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidin-1-yl)- 3,3-Dimethyl-1-oxobutane-2-amino)-2-oxyl)methyl)piperidin-1-yl)ethoxy)pyrido[4,3-d]pyrimidine- 4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate tert-butyl ester (14.5 mg, 10.94 μmol) was dissolved in tetrahydrofuran (4 mL), cooled to 0 ° C, added tetrabutyl ammonium fluoride (1M, 33.94 μL), stirred for 1 hour. Ethyl acetate (10 mL) was added to dilute the residue, washed with water (10 mL), the organic phase was collected, washed with saturated brine (150 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The title compound (12 mg, yield: 93.80%) was obtained.

MS m/z (ESI): 1169.5 [M+H] ⁺ .

The fifth step: (2S,4R)-1-(2S)-2-(1-(2-(4-(3,8-diazabicyclo[3.2.1]octane-3-yl)-7 -(8-ethynyl-7-fluoronaphthalen-1-yl)-8-fluoropyrido[4,3-d]pyrimidin-2-yl)oxy)ethyl)piperidin-4-yl)methoxy Base) acetamide)-3,3-dimethylbutyryl)-4-hydroxy-N-(S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrole Preparation of alkane-2-carboxamides

3-(7-(8-ethynyl-7-fluoronaphthalen-1-yl)-8-fluoro-2-(2-(S)-1-(2S,4R)-4-hydroxyl-2-( S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutane Alk-2-yl)amino)-2-oxoethoxy)piperidin-1-yl)ethoxy)pyrido[4,3-d]pyrimidin-4-yl)-3,8-diazepine Heterobicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester (12 mg, 10.26 μmol) was dissolved in dichloromethane (2 mL), added trifluoroacetic acid (310 μL), and stirred at 25° C. for 2 hours. Concentrate the reaction solution, dilute the residue with ethyl acetate (10 mL), adjust to pH = 8-9 with sodium bicarbonate solution, separate the layers, dry the organic phase with anhydrous sodium sulfate, concentrate, and pass the crude product through high performance liquid chromatography Purification gave the title compound (1.80 mg, yield: 16.41%).

MS m/z(ESI):1069.5[M+H] ⁺

¹ H-NMR (400MHz, CD ₃ OD) δ9.03(s, 1H), 8.87(d, J=4.0Hz, 1H), 8.54(s, 1H), 8.10(dd, J=12.0, 8.0Hz, 2H), 7.68-7.61(m, 2H), 7.47-7.36(m, 5H), 4.98(dd, J=16, 8Hz, 2H), 4.70-4.49(m, 8H), 4.40(d, J=24.0 Hz, 1H), 3.98(q, J=16.0Hz, 2H), 3.83(d, J=12.0Hz, 1H), 3.73(t, J=8.0Hz, 5H), 3.44(dd, J=16.0, 8.0 Hz,3H),3.09-3.00(m,2H),2.48-2.37(m,5H),2.24-2.17(m,1H),1.97-1.77(m,8H),1.56(d,J＝8.0Hz, 1H), 1.49-1.45(m, 4H), 1.37-1.25(m, 2H), 1.02(s, 9H).

Example 10: (2S,4R)-1-((S)-2-(2-((1-(2-((4-(3,8-diazabicyclo[3.2.1]octane- 3-yl)-7-(8-ethynyl-7-fluoro -3-Hydroxynaphthalen-1-yl)-8-fluoroquinazolin-2-yl)oxy)ethyl)piperidin-4-yl)methoxy)acetamido)-3,3-dimethyl butyryl)- Preparation of 4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (Compound 12)

The first step: 3-(2-(2-(4-((2-ethoxy-2-oxoethoxy)methyl)piperidin-1-yl)ethoxy)-8-fluoro- 7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalene-1-yl)quinazolin-4-yl)-3, Preparation of tert-butyl 8-diazabicyclo[3.2.1]octane-8-carboxylate

3-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)-2- (2-Oxoethoxy)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester (110.0mg, 137.3μmol) was dissolved in In dichloromethane (500.0 μL) and isopropanol (500.0 μL), add 2-(piperidin-4-ylmethoxy) ethyl acetate (41.4 mg, 206.0 μmol), sodium acetate trihydrate (93.4 mg, 686.5 μmol), and stirred for 15 minutes under an ice bath, then added 2-picoline borane (73.4 mg, 343.5 μmol), and stirred at 25° C. for 15 hours. The reaction solution was concentrated, and the residue was purified by silica gel column chromatography (eluent: dichloromethane/methanol=10/1, volume ratio) to obtain the title compound (90 mg, yield: 66.5%).

MS m/z (ESI): 986.0 [M+H] ⁺ .

The second step: 2-((1-(2-((4-(8-(tert-butoxycarbonyl)-3,8-diazabicyclo[3.2.1]octane-3-yl)-8 -Fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalene-1-yl)quinazolin-2-yl) Preparation of oxy)ethyl)piperidin-4-yl)methoxy)acetic acid

3-(2-(2-(4-((2-ethoxy-2-oxoethoxy)methyl)piperidin-1-yl)ethoxy)-8-fluoro-7-( 7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalene-1-yl)quinazolin-4-yl)-3,8-di Azabicyclo[3.2.1]octane-8-carboxylate tert-butyl ester (90.0mg, 91.3μmol) was dissolved in methanol (3mL) and water (0.7mL), and lithium hydroxide monohydrate (19.15mg, 456.5 μmol), stirred at 25°C for 2 hours. Add water (5mL) to the reaction solution, adjust the pH to 6 with 1M HCl aqueous solution, extract three times with ethyl acetate (5mL), collect the organic phase, wash with saturated brine (5mL) once, wash with anhydrous sodium sulfate The organic phase was dried and concentrated to obtain the title compound (20 mg, yield: 22.9%).

MS m/z (ESI): 958.0 [M+H] ⁺ .

The third step: 3-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalene-1-yl) -2-(2-(4-((2-(((S)-1-((2S,4R)-4-hydroxy-2-(((S)-1-(4-(4-methyl Thiazol-5-yl)phenyl)ethyl)))carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-2- Oxoethoxy)methyl)piperidin-1-yl)ethoxy)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert Preparation of butyl ester

2-((1-(2-((4-(8-(tert-butoxycarbonyl)-3,8-diazabicyclo[3.2.1]octane-3-yl)-8-fluoro- 7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalene-1-yl)quinazolin-2-yl)oxy) Ethyl)piperidin-4-yl)methoxy)acetic acid (20.0 mg, 20.88 μmol) and (2S,4R)-1-[(2S)-2-amino-3,3-dimethylbutyryl] -4-Hydroxy-N-[(1S)-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (11mg, 25μmol ) was dissolved in DMF (2mL), cooled to 0°C, N,N-diisopropylethylamine (10.8mg, 83.5μmol), 2-(7-azobenzotriazole)-N,N, N',N'-Tetramethyluronium hexafluorophosphate (15.9 mg, 41.8 μmol). Stir at 25°C for 3 hours. Add water (10 mL) to the reaction solution, extract three times with ethyl acetate (20 mL), wash the organic phase with saturated brine (20 mL), dry the organic phase with anhydrous sodium sulfate, and concentrate under reduced pressure, and pass the residue through a silica gel column Purification by chromatography (eluent: dichloromethane/methanol=10/1, volume ratio) gave the title compound (25 mg, yield: 86.5%).

MS m/z (ESI): 1384.0 [M+H] ⁺ .

The fourth step: 3-(7-(8-ethynyl-7-fluoro-3-(methoxymethoxy)naphthalene-1-yl)-8-fluoro-2-(2-(4-(( 2))-(((S)-1-((2S,4R)-4-hydroxy-2-(((S)-1-(4-(4-methylthiazol-5-yl)phenyl) Ethyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-2-oxoethoxy)methyl)piperidine Preparation of -1-yl)ethoxy)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester

3-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)-2- (2-(4-((2-(((S)-1-((2S,4R)-4-hydroxy-2-(((S)-1-(4-(4-methylthiazole-5 -yl)phenyl)ethyl)))carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-2-oxoethyl Oxygen)methyl)piperidin-1-yl)ethoxy)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester ( 25mg, 18.06μmol) was dissolved in DMF (1mL), then cesium fluoride (13.7mg, 90.3μmol) was added, protected with nitrogen, and stirred at 25°C for 2 hours. Water (10 mL) was added to the reaction solution, and a white solid was precipitated. The filter cake was collected by filtration and dried to obtain the title compound (20 mg, yield: 90.5%).

MS m/z (ESI): 1228.5 [M+H] ⁺ .

The fifth step: (2S,4R)-1-((S)-2-(2-((1-(2-((4-(3,8-diazabicyclo[3.2.1]octane- 3-yl)-7-(8-ethynyl-7-fluoro-3-hydroxynaphthalene-1-yl)-8-fluoroquinazolin-2-yl)oxy)ethyl)piperidin-4-yl )methoxy)acetylamino)-3,3-dimethylbutyryl)-4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl) Preparation of ethyl) pyrrolidine-2-carboxamide

3-(7-(8-ethynyl-7-fluoro-3-(methoxymethoxy)naphthalene-1-yl)-8-fluoro-2-(2-(4-((2)) -(((S)-1-((2S,4R)-4-hydroxy-2-(((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl) Carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-2-oxoethoxy)methyl)piperidine-1- Base)ethoxy)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester (20.00mg, 16.28μmol) was dissolved in dichloromethane (2 mL), trifluoroacetic acid (200 μL) was added, and stirred at 25° C. for 2 hours. Concentrate the reaction solution, dilute the residue with ethyl acetate (10 mL), adjust to pH = 8-9 with sodium bicarbonate solution, separate the layers, dry the organic phase with anhydrous sodium sulfate, concentrate, and pass the crude product through high performance liquid chromatography Purified by method to obtain the title compound (1 mg, yield: 5.7%).

MS m/z (ESI): 1084.0 [M+H] ⁺ .

¹ H-NMR (400MHz, CD ₃ OD) δ8.87-8.86 (m, 1H), 7.83-7.81 (m, 2H), 7.43-7.30 (m, 7H), 7.13-7.09 (m, 1H), 5.49 (s,5H),4.76-4.43(m,10H),3.96-3.48(m,7H),3.48-3.31(m,5H),2.69(s,1H),2.47-2.45(t,J＝8.0Hz ,3H), 2.21-1.93(m,12H), 1.33-1.28(m,4H), 1.02(s,9H).

Example 11: (2S,4R)-1-(2-(3-((1-(2-((4-(3,8-diazabicyclo[3.2.1]octane-3-yl) -7-(8-ethynyl-7-fluoro-3- Hydroxynaphthalene-1-yl)-8-fluoroquinazolin-2-yl)oxy)ethyl)piperidin-4-yl)methoxy)isoxazol-5-yl)-3-methylbutyl Acyl)-4-hydroxy Preparation of yl-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (compound 13)

The first step: 3-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalene-1-yl) -2-(2-(4-(((5-(1-((2S,4R)-4-hydroxy-2-(((S)-1-(4-(4-methylthiazole-5- Base) phenyl) ethyl)) carbamoyl) pyrrolidin-1-yl) -3-methyl-1-oxobutan-2-yl) isoxazol-3-yl) oxy) methyl Preparation of )piperidin-1-yl)ethoxy)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester

3-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)-2- (2-Oxoethoxy))quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester (55.0mg, 68.7μmol) In dichloromethane (500.0 μL) and isopropanol (500.0 μL), add (2S,4R)-4-hydroxy-1-(3-methyl-2-(3-(piperidin-4-ylmethyl Oxy)isoxazol-5-yl)butyryl)-N-(S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (61.3mg, 103.1μmol), sodium acetate trihydrate (46.7mg, 343.5μmol), stirred in ice bath for 15 minutes, then added 2-picoline borane (36.7mg, 343.5μmol), at 25°C Stir for 15 hours. The reaction solution was concentrated, and the residue was purified by silica gel column chromatography (eluent: dichloromethane/methanol=10/1, volume ratio) to obtain the title compound (40 mg, yield: 42.2%).

MS m/z (ESI): 1380.0 [M+H] ⁺ .

The second step: 3-(7-(8-ethynyl-7-fluoro-3-(methoxymethoxy)naphthalene-1-yl)-8-fluoro-2-(2-(4-(( (5-(1-((2S,4R)-4-hydroxy-2-(((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl )pyrrolidin-1-yl)-3-methyl-1-oxobutan-2-yl)isoxazol-3-yl)oxy)methyl)piperidin-1-yl)ethoxy) Preparation of tert-butyl quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate

3-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)-2- (2-(4-(((5-(1-((2S,4R)-4-hydroxy-2-(((S)-1-(4-(4-methylthiazol-5-yl)benzene Base) ethyl)) carbamoyl) pyrrolidin-1-yl)-3-methyl-1-oxobutan-2-yl) isoxazol-3-yl) oxy) methyl) piperidine -1-yl)ethoxy)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester (40mg, 28.99μmol) was dissolved in DMF (1 mL), then cesium fluoride (220 mg, 144.95 μmol) was added, protected with nitrogen, and stirred at 25° C. for 2 hours. Water (10 mL) was added to the reaction solution, and a white solid precipitated out. The filter cake was collected by filtration and dried to obtain the title compound (35 mg, yield: 98.6%).

MS m/z (ESI): 1225.5 [M+H] ⁺ .

The third step: (2S,4R)-1-(2-(3-((1-(2-((4-(3,8-diazabicyclo[3.2.1]octane-3-yl) -7-(8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluoroquinazolin-2-yl)oxy)ethyl)piperidin-4-yl)methoxy )isoxazol-5-yl)-3-methylbutyryl)-4-hydroxyl-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl ) Preparation of pyrrolidine-2-carboxamide

3-(7-(8-ethynyl-7-fluoro-3-(methoxymethoxy)naphthalen-1-yl)-8-fluoro-2-(2-(4-(((5- (1-((2S,4R)-4-Hydroxy-2-(((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidine -1-yl)-3-methyl-1-oxobutan-2-yl)isoxazol-3-yl)oxy)methyl)piperidin-1-yl)ethoxy)quinazoline -4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester (35.00 mg, 28.59 μmol) was dissolved in dichloromethane (2 mL), and trifluoroacetic acid ( 200 μL), stirred at 25°C for 2 hours. Concentrate the reaction solution, dilute the residue with ethyl acetate (10 mL), adjust the pH to 8-9 with sodium bicarbonate solution, separate the layers, dry the organic phase with anhydrous sodium sulfate, concentrate, and purify the crude product by high-performance liquid chromatography. The title compound (14.00 mg, yield: 45.3%) was obtained.

MS m/z (ESI): 1080.5 [M+H] ⁺ .

¹ H-NMR (400MHz, CD ₃ OD) δ8.87-8.84 (m, 1H), 7.86-7.77 (m, 2H), 7.38-7.27 (m, 6H), 7.08 (d, J = 4.0Hz, 1H ),5.86-5.80(m,1H),5.03-4.98(m,3H),4.66-4.44(m,10H),3.92-3.76(m,3H),3.70-3.48(m,4H),3.10(dd ,J＝4.0,4.0Hz,2H),2.68(s,1H),2.46-2.37(m,6H),2.21-1.76(m,9H),1.57-1.16(m,8H),1.05-0.88(m ,6H).

Biological example:

Test Example 1: Surface Plasmon Resonance (SPR) Binding Test of Compounds and His-KRAS (G12D&Q61H) Proteins

●Test system and parameters

Protein: His-KRAS (G12D&Q61H) (Shenzhou);

Buffer system:

1) Protein immobilization buffer: 10mM HEPES, 150mM NaCl, 0.05% Tween-20 and 10μM GDP;

2) Running buffer A: 10mM HEPES, 150mM NaCl, 0.05% Tween-20 and 10μM GDP;

3) Running buffer B: 10mM HEPES, 150mM NaCl, 0.05% Tween-20, 10μM GDP and 1% DMSO;

Compound concentration: 50 μM to 0.781 μM in running buffer B, two-fold serial dilution, the final concentration of DMSO is 1%;

Binding and dissociation time: 120s on/200s off;

Flow rate: 50μL/min;

Instrument: Biacore T200 (GE Healthcare);

Chip: S series NTA sensor chip.

●Test procedure

(1) Protein immobilization

1) Temperature setting: set the temperature of the instrument to 25°C.

2) Insert the S series NTA sensor chip, and then execute the "Prime" command.

3) Start the manual program and set the flow rate to 10 μL/min.

4) Select flow cell 2, and then automatically inject 350mM EDTA (pH 8.0) solution and 50mM NaOH solution in sequence (injection speed 60μL/min, injection time 60s, each repeated twice) to rinse the chip surface.

5) Chip surface activation: select flow cell 2, automatically inject 10 mM NiCl ₂ , the flow rate is 4 μL/min, and the injection time is 500 s.

6) Dilute His-KRAS (G12D&Q61H) to 40 μg/mL with protein immobilization buffer, and then automatically inject 120 μL of His-KRAS (G12D&Q61H) (injection speed is 4 μL/min, injection time is 1150s) to extract His- KRAS (G12D&Q61H) protein is immobilized on the surface of flow cell 2.

7) Adjust the flow rate back to 10 μL/min, and balance the surface of the chip until the baseline is basically stable (baseline drop rate <2RU/min).

(2) Sample testing

Use the method mode to write programs in the Biacore T200 control software. The temperature was set at 25°C and the flow rate was set at 50 μL/min. The program first performs 8 injections of running buffer B, and then sets the automatic injection according to the order of compound concentration from small to large, so that the compound flows over the immobilized protein surface on the chip and binds to the chip. Set flow cell 1 as the reference channel, and set 0 concentration as the built-in blank control. The needle was rinsed with 50% DMSO after each cycle. A 1% DMSO standard curve was used to correct for the signal value effect of DMSO.

●Data processing

The binding and dissociation signals of the compound and the protein were recorded in real time, and the signal values of the reference channel and the built-in blank control were subtracted (double subtraction) during data processing (Biacore T200 evaluation software). The sensorgram of the double-subtracted signal values of the reference channel and the built-in blank control was fitted by Kinetics or steady-state Affinity (1:1) mode. The K _D value (K _d /K _a ) is used to characterize the affinity of the compound to the protein, where K _d is the dissociation rate constant and K _a is the association rate constant.

●Test results

The affinity between the compound and the His-KRAS (G12D&Q61H) protein was determined according to the above method, and the results are shown in Table 1:

Table 1. Affinity test results of compounds and His-KRAS (G12D&Q61H)

compound K _D (nM) 2 191 3 329 7 12

●Test conclusion

In the binding test between the compound and His-KRAS (G12D&Q61H), the compound of the present invention shows a strong affinity.

Test Example 2: KRAS-G12D (guanine-5'-triphosphate (GTP) hydrolase G12D mutant)/SOS1 protein binding in vitro Inhibitory Activity Test

●Test system and parameters

KRAS-G12D/SOS1 Binding Assay Kit (Cisbio);

Proteins: Tag1-SOS1 and Tag2-KRAS G12D;

Tags: Anti tag1 tb ³⁺ and Anti tag2 XL665;

Buffer: dilution buffer and detection buffer;

Activating nucleotide: GTP (Sigma);

KRAS-G12D/SOS1 binding assay kit components: Tag1-SOS1, Tag2-KRAS G12D, Anti tag1 tb ³⁺ and Anti tag2 XL665 are all 1ⅹ;

GTP: 10000nM;

Compound and KRAS-G12D/SOS1 protein pre-incubation binding time: 15 minutes at room temperature;

Reaction time of protein and label: 4 ℃ sealed incubation for 3 hours or extended to overnight;

Microplate reader: BMG PHERAstar Fluorescence, homogeneous time-resolved fluorescence (HTRF) method, excitation wavelength 337nm, emission wavelength 665nm and 620nm.

●Test procedure

For protein binding, test compounds were incubated with a mixture of Tag1-SOS1, Tag2-KRAS G12D, and GTP in dilution buffer for 15 minutes at room temperature. Add labels diluted in Assay Buffer to the reaction plate and incubate at 4°C for 3 hr or extend to overnight. Put the reaction plate into the microplate reader, and use the HTRF method to read the signal value of each well in the plate.

●Data processing

The vehicle group (containing 1ⅹTag1-SOS1, 1ⅹTag2-KRAS G12D, Anti tag1 tb ³⁺ and Anti tag2 XL665, 1% DMSO) was used as negative control, and the reaction buffer group (containing 1ⅹTag1-SOS1, Anti Tag1 tb ³⁺ and Anti tag2 XL665, 1% DMSO) was used as a blank control, and the relative inhibitory activity of each concentration group was calculated, and the inhibition rate=100%-(test group-blank group)/(vehicle group-blank group)*100%. The curve was fitted according to the four-parameter model, and the half-maximal inhibitory concentration (IC ₅₀ ) of the compound was calculated.

●Test results

The KRAS-G12D/SOS1 in vitro protein binding inhibitory activity of the compounds was determined according to the above method, and the results are shown in Table 2.

Table 2. KRAS-G12D/SOS1 protein binding inhibitory activity test results in vitro

Compound number IC ₅₀ (nM) 1 410 2 61 3 153 4 32 5 49 6 43 7 13

●Test conclusion

In the KRAS-G12D/SOS1 protein binding inhibitory activity test in vitro, the compound of the present invention exhibits strong inhibitory binding activity.

Test Example 3: Compound and KRAS-G12D (guanine-5'-triphosphate (GTP) hydrolase G12D mutant)/VHL in vitro Enzymatic Ternary Complex Binding Activity Assay

●Test system and parameters

protein:

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

Human VHL recombinant protein (Abnova): 300nM

Label:

Anti-6xHis AlphaLISA receptor beads (PerkinElmer): 20 μg/mL

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

Buffer: AlphaLISA 10X buffer (PerkinElmer)

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

●Test procedure

The compound to be tested was pre-incubated with KRAS/K-Ras (G12D&Q61H) protein and human VHL recombinant protein at 23° C. for 60 minutes to bind. Add acceptor beads and donor beads and incubate at 23 °C for 60 min in the dark. Put the reaction plate into the microplate reader, and use the AlphaLISA method to read the signal value of each well in the plate.

●Data processing

The wells where the samples were added (containing samples of different dilution concentrations, KRAS/K-Ras (G12D&Q61H) protein, human VHL recombinant protein, anti-6xHis AlphaLISA acceptor microspheres and α-glutathione donor microspheres) were used as the test group , with the vehicle group (containing KRAS/K-Ras (G12D&Q61H) protein, human VHL recombinant protein, anti-6xHis AlphaLISA acceptor microspheres and α-glutathione donor microspheres, 1% DMSO) as negative control, calculate the For the relative activation activity of the compounds, the standard curve was fitted using the four-parameter equation of SigmaPlot 12.5 software, and the EC ₅₀ was calculated.

Test Example 4: KRAS-G12D Protein Degradation Test in Cells by Compounds

●Test system and parameters:

Cells: Human colon cancer cell line GP2D or human metastatic pancreatic adenocarcinoma AsPC-1 cells

Protein quantification: Pierce ^TM BCA protein detection kit (Thermofisher)

Protein electrophoresis: electrophoresis tank (Thermo) 180V constant voltage electrophoresis

Protein transfer: transfer tank (BIO-RAD) 300mA constant flow transfer membrane

●Test procedure

(1) Compound treated cells

The cells were plated in a six-well plate with a cell volume of 1.5ⅹ10 ⁶ cells per well. Adhesive culture was carried out overnight, and compounds (concentrations of 0.25, 2.5, and 25 μM) were added for treatment for 24 hours.

(2) Sample processing

1) Discard the medium, add 2ml PBS, and wash the cells. Discard the washing solution, add an appropriate amount of cell lysate (#9803), place it on ice for 30 minutes, then centrifuge at 12,000 rpm for 10 minutes at 4°C, and remove the supernatant.

2) Referring to the instructions of the BCA kit, take 40 μL of standard concentration protein (2mg/ml) and cell lysate for 1:1 gradient dilution to obtain standard concentration protein dilutions of 1, 0.5, 0.25, 0.125, and 0.0625 mg/ml. Take 10 μL of the supernatant obtained in step 1) and the protein dilution of standard concentration and add them to the sample wells of a 96-well plate, and add 200 μL of A and B reagent mixture (50:1) to each well. Shake at 37°C for 30 minutes, and then detect the OD value at 562nm. Fit the standard curve of the standard concentration protein dilution, calculate the protein concentration in the supernatant obtained in step 1) according to the curve, and through appropriate dilution, the protein concentration in the supernatant with the lowest protein concentration shall prevail. The protein concentration of each remaining supernatant was adjusted to this concentration.

3) Add 5X SDS-PAGE protein loading buffer (Beiyuntian, P0015L) to the sample, and denature the protein at 100°C for 10 minutes.

(3) Protein separation transfer and development

1) Use NuPAGE 4-12% gradient protein precast gel (Thermofisher, NP0336BOX) to separate protein samples by 180V constant voltage electrophoresis. The gel was transferred to the membrane in a constant flow transfer tank for 90 minutes, and blocked in 5% skimmed milk powder.

2) Use KRAS-G12D antibody (CST, 14429) and α-Tubulin antibody (CST, 2125S) to dilute according to the dilution ratio of the antibody instructions, and then incubate with the membrane overnight at 4°C, wash with TBST and use goat anti-rabbit HRP secondary antibody (CST, 7074) was diluted according to the dilution ratio of the antibody manual, and then incubated with the membrane at room temperature for 1 h, and washed with TBST.

3) Soak the membrane in ECL luminescence solution (Millipore\WBKLS0500), and use a Bio-rad gel imager to collect fluorescence signals.

●Data processing

Use Image J software to quantify the result pictures, and calculate the degradation rate (KRAS-G12D) by comparing with the control group without compound, the calculation formula is as follows:

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

●Test results

The degradation effect of the compound on KRAS-G12D protein in cells was determined according to the above method, and the results are shown in Table 3 and Table 4.

Table 3. Degradation rate of compounds on KRAS-G12D protein in human colon cancer cell GP2D cells

Compound number Degradation rate (2.5μM) Degradation rate (25μM) 2 61.88% 75.08%

Table 4. Degradation rate of compounds on KRAS-G12D protein in human metastatic pancreatic adenocarcinoma AsPC-1 cells

Compound number Degradation rate (2.5μM) 7 92.06%

●Test conclusion

The compound of the present invention exhibits strong degrading activity on target protein in cells. Test Example 5: Cell Proliferation Inhibitory Activity Test of Compounds

●Test system and procedures

AsPC-1, GP2D, Panc 04.03 and AGS cells were cultured in RPMI1640+10% fetal bovine serum (FBS), DMEM+10% FBS, RPMI1640+15% FBS+10 μg/mL insulin, and Ham's F12 medium, respectively. Cultured in a carbon dioxide incubator at 37°C. Cells were plated in a 96-well flat-bottomed cell culture plate, with 2000 cells per well, and cultured overnight. The next day, compound dilutions were added. The compound was diluted to a maximum concentration of 100 μM, and 9 concentration points were diluted in a 4-fold concentration gradient, or the compound was diluted to a maximum concentration of 3 μM, and a 3-fold concentration gradient was diluted to 9 concentration points, and DMSO was used as a control. The final concentration of DMSO in the cell culture medium was 1%.

试剂(Promega公司)检测，化合物与GP2D、Panc 04.03和AGS细胞在37℃的二氧化碳培养箱中孵育6天后使用CELL

试剂(Promega公司)检测。未处理细胞基线在加入化合物当天检测。 Compounds were incubated with AsPC-1 cells in a carbon dioxide incubator at 37°C for 7 days before using CELL

Reagent (Promega Company) detection, the compound was incubated with GP2D, Panc 04.03 and AGS cells in a carbon dioxide incubator at 37°C for 6 days and then used in CELL

Reagent (Promega company) detection. The baseline of untreated cells was measured on the day of compound addition.

●Data processing

The inhibitory activity of the compound is calculated by the following formula: inhibition rate=100%-(test group/vehicle group)*100%. Using the SigmaPlot 12.5 software four-parameter model to fit the curve, the half inhibitory concentration (IC ₅₀ ) of the compound on cell proliferation was calculated.

●Test results

Determination of compound's proliferation inhibitory activity on KRAS ^G12D mutant cells according to the above method, the results are shown in Table 5

Table 5. Proliferation inhibitory activity of compounds on KRAS ^G12D mutant cells

●Test conclusion

The compound of the present invention exhibits strong proliferation inhibitory activity on KRAS ^G12D mutant cells.

The above examples do not limit the scope of the invention in any way. Various modifications of the invention, in addition to those described herein, will be apparent to those skilled in the art from 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 publications, is hereby incorporated by reference in its entirety.

Claims (15)

A compound, or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite or prodrug thereof, which compound has The structure of formula (I):

in:

is a single or double bond; When between X ¹ and X ²

When it is a single bond, X ¹ is selected from C(R ¹ ) ₂ and C(O); and X ² is selected from N and CR ¹ ; When between X ¹ and X ²

When it is a double bond, X ¹ is selected from CR ¹ and N; and X ² is C; When between X ³ and X ⁴

When it is a single bond, X ³ and X ⁴ are each independently selected from C(R ¹ ) ₂ , NR ⁸ and C(O); When between X ³ and X ⁴

When it is a double bond, X ³ and X ⁴ are each independently selected from CR ¹ and N; Each R ¹ is independently selected from hydrogen, halogen, C _1-6 alkyl, C _1-6 alkenyl, C _3-6 cycloalkyl, C _1-6 haloalkyl, -OC _1-6 alkyl, - OC _1-6 haloalkyl and -OC _3-6 cycloalkyl; or two R ¹ on the same carbon atom form carbonyl, C _3-6 cycloalkyl or 3-6 membered heterocycle with the carbon atom they are connected to base; R ⁸ is independently selected from hydrogen, C _1-3 alkyl, C _3-6 cycloalkyl and C _1-3 haloalkyl; R ² is selected from C _6-10 aryl or 5-10 membered heteroaryl, each of said aryl and heteroaryl is optionally substituted by one or more R ³ ; When present, each R ³ is independently selected from halogen, hydroxy, cyano, C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl, C _1-6 haloalkyl, -C _{1 -6} alkyl-OH, -OC _1-6 alkyl, C _3-6 cycloalkyl, 3-10 membered heterocyclyl and 5-10 membered heteroaryl, the alkyl, alkenyl, alkynyl, Cycloalkyl, heterocyclyl and heteroaryl are each optionally substituted by one or more substituents selected from halogen, hydroxy, cyano, C _1-6 alkyl and C _1-6 haloalkyl; L is -L ¹ -L ² -L ³ -L ⁴ -L ⁵ -L ⁶ -; L ¹ is selected from -O-, -S-, -NR ^1a -, C _1-6 alkylene, C _3-6 cycloalkylene and 3-10 membered heterocyclylene, said alkylene, alkylene Cycloalkyl and heterocyclylene are each optionally substituted by one or more substituents selected from halogen, hydroxy, cyano, C _1-6 alkyl and C _1-6 haloalkyl; L ² is a covalent bond, or selected from -O-, -NR ^2a -, C _1-6 alkylene, -OC _1-6 alkylene, C _1-6 alkylene-O-, C _{3- 6} cycloalkylene groups and 3-10 membered heterocyclylene groups, each of the alkylene, cycloalkylene and heterocyclylene groups is optionally selected from one or more halogen, hydroxyl, cyano, C ₁ Substituents of _-6 alkyl and C _1-6 haloalkyl; L ³ is a covalent bond, or selected from -O-, -NR ^3a -, C _1-6 alkylene, -OC _1-6 alkylene, C _1-6 alkylene-O-, C _{3- 6} cycloalkylene groups and 3-10 membered heterocyclylene groups, each of the alkylene, cycloalkylene and heterocyclylene groups is optionally selected from one or more of halogen, hydroxyl, cyano, C ₁ Substituents of _-6 alkyl and C _1-6 haloalkyl; L ⁴ is a covalent bond, or is selected from -O-, -NR ^4a -, C _1-6 alkylene, -OC _1-6 alkylene, C _1-6 alkylene-O-, C _{3- 6} cycloalkylene groups and 3-10 membered heterocyclylene groups, each of the alkylene, cycloalkylene and heterocyclylene groups is optionally selected from one or more halogen, hydroxyl, cyano, C ₁ Substituents of _-6 alkyl and C _1-6 haloalkyl; L ⁵ is a covalent bond, or selected from -O-, -NR ^5a -, C _1-6 alkylene, -OC _1-6 alkylene-, C _1-6 alkylene-O-, C _{3 -6} cycloalkylene and 3-10 membered heterocyclylene, said alkylene, cycloalkylene and heterocyclylene are each optionally replaced by one or more selected from halogen, hydroxyl, cyano, C Substituents of _1-6 alkyl and C _1-6 haloalkyl; L ⁶ is selected from -O-, -S-, -NR ^6a -, C _1-6 alkylene, -OC _1-6 alkylene-, C _1-6 alkylene-O-, C _3-6 Cycloalkylene, -C(O)-, -C(O)-N(R ^6a )- and 3-10 membered heterocyclylene, each of the alkylene, cycloalkylene and heterocyclylene optionally substituted by one or more substituents selected from halogen, hydroxyl, cyano, C _1-6 alkyl and C _1-6 haloalkyl; R ^1a , R ^2a , R ^3a , R ^4a , R ^5a and R ^6a are each independently selected from hydrogen and C _1-6 alkyl; M is a small molecule ligand of E3 ubiquitin ligase VHL, preferably its structure is shown in the following formula:

in A is selected from -NH-, -EC(=O)-NR ^7a -, -E-NR ^7a -C(=O)-, -EC(=O)- and 5-10 membered heteroarylene, said Heteroarylene is optionally substituted by one or more substituents selected from halogen, hydroxy, cyano, C _1-6 alkyl and C _1-6 haloalkyl; E is selected from C _1-6 alkylene, C _3-6 cycloalkylene and 3-10 membered heterocyclylene, each of which is optionally replaced by one Or more substituents selected from halogen, hydroxyl, cyano, C _1-6 alkyl and C _1-6 haloalkyl; R ^7a is selected from hydrogen and C _1-6 alkyl; R ⁴ is selected from hydrogen, C _1-6 alkyl, C _3-6 cycloalkyl and 3-12 membered heterocyclyl; R ^is selected from hydrogen, C _1-6 alkyl, C _3-6 cycloalkyl, 3-12 membered heterocyclyl, each of which is optionally replaced by one or more Substituents selected from halogen, hydroxyl, cyano, C _1-6 alkyl and C _1-6 haloalkyl; R is selected from hydrogen, halogen, hydroxyl, C ^1-6 _alkyl , C _2-6 alkenyl and C _2-6 alkynyl; and

Indicates the junction site. The compound of claim 1 or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotope-labeled compound, metabolite or precursor thereof Drug, wherein said compound has the structure of formula (IA), (IB) or (IC):

wherein R ¹ , R ² , L and M are as defined in claim 1 , and n is 0, 1, 2, 3 or 4 for R ¹ other than hydrogen. The compound of claim 1 or 2 or its pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotope-labeled compound, metabolite Or a prodrug, wherein each R ¹ is independently selected from hydrogen and halogen such as fluorine or chlorine. The compound described in any one of claims 1-3 or its pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotope-labeled Compounds, metabolites or prodrugs, wherein R ² is naphthyl optionally substituted by 1-3 R ³ , and each R ³ is independently selected from F, Cl, Br, I, methyl, ethyl, iso Propyl, ethynyl and hydroxyl, preferably R2 is ⁸ -Cl-naphthalen-1-yl, 3-OH-naphthalen-1-yl, 8-ethynyl-7-F-3-OH-naphthalen-1-yl Or 8-ethynyl-7-F-naphthalen-1-yl. The compound according to any one of claims 1-4 or its pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotope-labeled Compound, metabolite or prodrug, wherein L ¹ is -O-. The compound described in any one of claims 1-5 or its pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotope-labeled Compounds, metabolites or prodrugs, wherein L ² is C _1-6 alkylene, preferably -CH ₂ - or -CH ₂ -CH ₂ -. The compound described in any one of claims 1-6 or its pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotope-labeled Compounds, metabolites or prodrugs, wherein L ³ is a 3-6 membered heterocyclic group containing an N atom, preferably a 5-6 membered heterocyclic group containing an N atom, preferably

The compound according to any one of claims 1-7 or its pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotope-labeled Compound, metabolite or prodrug, wherein L ⁴ and L ⁵ are each independently selected from covalent bond, C _1-6 alkylene, -OC _1-6 alkylene- and C _1-6 alkylene-O -, preferably -L ⁴ -L ⁵ - is -CH ₂ -CH ₂ -CH ₂ -O-CH ₂ -CH ₂ -, -CH ₂ -CH ₂ -CH ₂ -CH ₂ -, -CH ₂ -CH _{2 -CH2} - _CH2 -CH2-, _-CH2 - _CH2 - _CH2 - _CH2 - _{CH2 -} CH2-, _{-CH2 -} O _- CH2-, or _-CH2- . The compound described in any one of claims 1-8 or its pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotope-labeled Compounds, metabolites or prodrugs, wherein L ⁶ is -C(O)- or -O-. The compound according to any one of claims 1-9 or its pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotope-labeled Compounds, metabolites or prodrugs, wherein A is -NH-; R ⁴ is C _1-6 alkyl, preferably tert-butyl; and R ⁵ and R ⁶ are both hydrogen. The compound according to any one of claims 1-9 or its pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotope-labeled Compounds, metabolites or prodrugs, wherein A is a 5-10 membered heteroarylene group, preferably a 5-6 membered heteroarylene group, preferably

R ⁴ is C _1-6 alkyl, preferably isopropyl; R ⁵ is C _1-6 alkyl, preferably methyl; and R ⁶ is hydrogen. The compound described in any one of claims 1-11 or its pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotope-labeled Compound, metabolite or prodrug, wherein said compound is selected from:

A pharmaceutical composition comprising a preventive and/or therapeutically effective amount of the compound of any one of claims 1-12 or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorphic form thereof Compounds, solvates, N-oxides, isotope-labeled compounds, metabolites or prodrugs, and one or more pharmaceutically acceptable carriers. The compound described in any one of claims 1-12 or its pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotope-labeled Use of the compound, metabolite or prodrug or the composition described in claim 13 in the preparation of a medicament for preventing and/or treating KRAS G12D-mediated related diseases. The method for preparing the compound described in claim 2, it is shown in following route 1, route 2, route 3 or route 4: Route 1

in R ¹ , R ² , n, L and M are as defined in any one of claims 1-9; LG ^1a , LG ^2a , LG ^3a and X each independently represent a leaving group such as halogen, triflate, thiomethyl, methylsulfoxide, methylsulfone, methoxy or ethoxy ; PG ^1a and PG ^2a each independently represent a protecting group, such as benzyloxycarbonyl (Cbz), tert-butoxycarbonyl (Boc), methyl or benzyl; The method comprises the steps of: (1) reacting Compound IA-1 with Compound IA-2 to obtain Compound IA-3; (2) reacting Compound IA-3 with Compound IA-4 to obtain Compound IA-5; (3) subjecting compound IA-5 to a deprotection reaction to obtain compound IA-6; (4) making compound IA-6 and compound IA-7 carry out coupling reaction to obtain compound IA-8; (5) reacting Compound IA-8 with Compound IA-9 to obtain Compound IA-10; and (6) subjecting compound IA-10 to a deprotection reaction to obtain compound IA; route 2

in R ¹ , R ² , L and M are as defined in any one of claims 1-9; LG ^b , LG ^1b , LG ^2b , LG ^3b and LG ^4b each independently represent a leaving group such as halogen, triflate, thiomethyl, methylsulfoxide, methylsulfone, boronic acid, boron ester, tributyltin, methoxy or ethoxy; PG ^1b represents a protecting group, such as benzyloxycarbonyl (Cbz) or tert-butoxycarbonyl (Boc); The method comprises the steps of: 1) reacting compound IB-1 with compound IB-2 to obtain compound IB-3; 2) reacting compound IB-3 with compound IB-4 to obtain compound IB-5; 3) make compound IB-5 and compound IB-6 carry out coupling reaction to obtain compound IB-7; 4) reacting compound IB-7 with compound IB-8 to obtain compound IB-9; and 5) subjecting compound IB-9 to a deprotection reaction to obtain compound IB; Route 3

in wherein R ¹ , R ² , L, L ¹ , L ² , L ³ , L ⁴ , L ⁵ , L ⁶ and M are as defined above; LG ^b , LG ^1b , LG ^2b and LG ^4b each independently represent a leaving group such as halogen, triflate, thiomethyl, methylsulfoxide, methylsulfone, boronic acid, borate ester, Tributyltin, methoxy or ethoxy; PG ^1b represents a protecting group, such as benzyloxycarbonyl (Cbz) or tert-butoxycarbonyl (Boc); The method comprises the steps of: 1) reacting compound IB-1 with compound IB-2 to obtain compound IB-3; 2) reacting compound IB-3 with compound IB-10 to obtain compound IB-11; 3) performing a coupling reaction between compound IB-11 and compound IB-6 to obtain compound IB-12; 4) subjecting compound IB-12 to an oxidation reaction to obtain compound IB-13; 5) reacting compound IB-13 with compound IB-14 to obtain compound IB-15; and 6) subjecting compound IB-15 to a deprotection reaction to obtain compound IB; Route 4

in wherein R ¹ , R ² , L, L ¹ , L ² , L ³ , L ⁴ , L ⁵ , L ⁶ , n and M are as defined above; LG ^c , LG ^1c , LG ^2c and LG ^3c each independently represent a leaving group such as halogen, triflate, thiomethyl, methylsulfoxide, methylsulfone, boronic acid, borate ester, Tributyltin, methoxy or ethoxy; PG ^1c represents a protecting group, such as benzyloxycarbonyl (Cbz) or tert-butoxycarbonyl (Boc); The method comprises the steps of: 1) reacting compound IC-1 with compound IC-2 to obtain compound IC-3; 2) reacting compound IC-3 with compound IC-4 to obtain compound IC-5; 3) performing a coupling reaction between compound IC-5 and compound IC-6 to obtain compound IC-7; 4) subjecting compound IC-7 to an oxidation reaction to obtain compound IC-8; 5) reacting compound IC-8 with compound IC-9 to obtain compound IC-10; and 6) Subjecting compound IC-10 to a deprotection reaction to obtain compound IC.