CN115850267A

CN115850267A - Bridged ring compound, preparation method and application thereof

Info

Publication number: CN115850267A
Application number: CN202211115052.4A
Authority: CN
Inventors: 刘金明; 唐建川; 任云; 何婷; 康熙伟; 刘叶; 袁晓曦; 周俊鹏; 刘谦; 田强; 宋宏梅; 葛均友; 王晶翼
Original assignee: Sichuan Kelun Biotech Biopharmaceutical Co Ltd
Current assignee: Sichuan Kelun Biotech Biopharmaceutical Co Ltd
Priority date: 2021-09-24
Filing date: 2022-09-14
Publication date: 2023-03-28

Abstract

The invention relates to a bridged ring compound, a preparation method and application thereof. In particular, the invention relates to a compound with a structure of formula (I), and the compoundHas strong inhibiting effect on the target KRAS G12D, can be used for preventing and/or treating KRAS G12D-mediated related diseases, and has better tumor treatment effect.

Description

Bridged ring compound, preparation method and application thereof

Technical Field

The invention belongs to the field of pharmaceutical chemistry, and particularly relates to a bridged ring compound, a preparation method thereof, a pharmaceutical composition containing the bridged ring compound and application of the bridged ring compound.

Background

The RAS family is a class of guanine nucleotide binding proteins with gtpase activity. As a molecular switch, RAS can realize the conversion between GDP-bound inactive state and GTP-bound active state, transmit upstream signals received by cells to downstream multiple signal pathways, regulate protein synthesis, gene transcription, cell growth, differentiation, apoptosis, migration, and the like.

In the field of cancer research, the RAS gene is one of the most prevalent proto-oncogenes in human cancers. RAS mutations result in the persistent activation of downstream signaling pathways, promoting tumorigenesis. The RAS family includes HRAS, NRAS and KRAS, where about 85% of RAS mutations occur in all tumor types. In KRAS mutant tumors, the GTPase activity of KRAS itself is reduced, and KRAS maintains the activity state continuously. KRAS mutations are closely related to the development of lung, pancreatic and colorectal cancer, and the mutation of glycine 12 to aspartic acid (KRAS G12D) occurs most frequently (about 34%) on KRAS.

The research and development aiming at KRAS G12D small-molecule inhibitors is one of the difficulties in the field of medicine. BI-2852 of Boringer Vargham company induces KRAS G12D to form a dimer by using a molecular glue principle, and blocks the interaction between KRAS and downstream protein; the Revolition company induces KRAS G12D protein to form a ternary complex by using an mTOR inhibitor, and blocks the interaction of KRAS and downstream effector protein; the company Miratii, WO2021041671A1, discloses a novel class of KRAS G12D inhibitors, the mechanism of action of which is not disclosed.

Although KRAS has been an excellent target for tumor therapy, there is no clinically validated drug targeting KRAS G12D. Therefore, the development of KRAS G12D targeted inhibitors with novel structures, good biological activities and high drug-forming properties is urgently needed in the field.

Disclosure of Invention

On one hand, the invention provides a bridged ring compound which has a strong inhibition effect on a target KRAS G12D, so that the compound has a better tumor treatment effect. The compounds of the invention also have a number of advantageous properties, such as good physicochemical properties (e.g. solubility, physical and/or chemical stability) and good safety.

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

wherein,

ring A is 5-7 membered heterocycle, 5-6 membered heteroaromatic ring, C _5-7 Cycloalkyl, phenyl, 8-11 membered spiroheterocycle or C _8-11 Spirocycloalkyl, the A ring being optionally substituted by one or more R ³ Substitution;

X ¹ selected from N and CR ⁴ ；

L ¹ Selected from the group consisting of covalent bonds, -O-) -C (O) -, -S-and-NR ⁵ -；

L ² Selected from the group consisting of a covalent bond, -O-, and-C (O) -;

R ¹ selected from hydrogen, C _1-6 Alkyl radical, C _3-10 Cycloalkyl and 3-12 membered heterocyclyl, each optionally substituted with one or more R ⁶ Substitution;

R ² is selected from C _6-10 Aryl and 5-10 membered heteroaryl, each optionally substituted with one or more R ⁷ Substitution;

R ³ selected from hydrogen, halogen, hydroxy, cyano,C _1-6 Alkyl radical, C _1-6 Haloalkyl, C _2-6 Alkenyl radical, C _2-6 Alkynyl, -O-C _1-6 Alkyl, -O-C _3-6 Cycloalkyl, -O-C _2-6 Alkenyl, -O-C _2-6 Alkynyl, C _3-10 Cycloalkyl and 3-12 membered heterocyclyl; or two R of the same carbon atom ³ And the carbon atoms to which they are both attached form a carbonyl group;

R ⁴ selected from hydrogen, halogen, cyano, C _1-6 Alkyl and C _1-6 A haloalkyl group;

R ⁵ selected from hydrogen and C _1-6 An alkyl group;

R ⁶ selected from hydrogen, halogen, cyano, -OR ⁹ 、-O-C(O)-N(R ⁹ )R ¹⁰ 、-O-C(O)-R ⁹ 、-N(R ⁹ )R ¹⁰ 、-N(R ¹⁰ )-C(O)-R ⁹ 、-N(R ¹⁰ )-C(O)-N(R ⁹ )R ¹⁰ 、-N(R ¹⁰ )-C(O)-OR ⁹ 、-C(O)-R ⁹ 、-C(O)-OR ¹⁰ 、-C(O)-N(R ⁹ )R ¹⁰ 、C _1-6 Alkyl radical, C _1-6 Haloalkyl, C _3-10 Cycloalkyl radical, C _6-10 Aryl, 5-10 membered heteroaryl, and 3-12 membered heterocyclyl, each of which is optionally substituted with one or more substituents independently selected from the group consisting of: hydrogen, halogen, hydroxy, amino, cyano, C _1-6 Alkyl radical, C _1-6 Haloalkyl, -C _1-6 alkylene-R ¹¹ 、-C _1-6 alkylene-OR ¹¹ and-C _1-6 alkylene-NR ¹¹ R ¹² ；

R ⁷ Selected from hydrogen, halogen, hydroxy, cyano, C _1-6 Alkyl radical, C _2-6 Alkenyl radical, C _2-6 Alkynyl, -O-C _1-6 Alkyl, -O-C _3-6 Cycloalkyl radical, C _3-10 Cycloalkyl, 3-12 membered heterocyclyl and 5-10 membered heteroaryl, each optionally substituted with one or more substituents selected from hydrogen, halogen, hydroxy, cyano, C _1-6 Alkyl and C _1-6 Substituted with a haloalkyl; or,

R ³ and R ⁷ In conjunction withAdjacent atoms together forming C _5-7 Cycloalkyl or 5-7 membered heterocycle;

R ⁸ selected from halogen, hydroxy, cyano, C _1-6 Alkyl, -C _1-6 alkylene-OH, -C _1-6 Alkylene-cyano and C _1-6 A haloalkyl group;

R ⁹ selected from hydrogen, C _1-6 Alkyl radical, C _1-6 Haloalkyl, -C _1-6 alkylene-O-C _1-6 Alkyl radical, C _3-10 Cycloalkyl, 3-12 membered heterocyclyl, each of which is optionally substituted with one or more substituents selected from hydrogen, halogen, hydroxy, cyano, C _1-6 Alkyl radical, C _1-6 Substituted with a haloalkyl;

R ¹⁰ selected from hydrogen and C _1-6 An alkyl group;

R ¹¹ selected from hydrogen, C _1-6 Alkyl radical, C _1-6 Haloalkyl, C _3-10 Cycloalkyl, 3-12 membered heterocyclyl, each of which is optionally substituted with one or more substituents selected from hydrogen, halogen, hydroxy, cyano, C _1-6 Alkyl radical, C _1-6 Substituted with a substituent of haloalkyl;

R ¹² selected from hydrogen and C _1-6 An alkyl group;

n is 0, 1 or 2;

o, p, q, r and s are each independently selected from 0, 1,2 or 3, and o and r are not simultaneously 0and p and q are not simultaneously 0;

provided that when

Is->

Is->

-L ¹ -R ¹ Is->

When is-L ² -R ² Is not->

In another aspect, the invention provides a pharmaceutical composition comprising a prophylactically or therapeutically effective amount of a compound of the invention, or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound or metabolite thereof, and one or more pharmaceutically acceptable carriers.

In another aspect, the invention provides a kit comprising a compound of the invention, or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound or metabolite thereof, or a pharmaceutical composition of 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 metabolite thereof, or a pharmaceutical composition of the present invention, for use in the inhibition of KRAS G12D.

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 metabolite thereof, or a pharmaceutical composition of the present invention, for use in the prevention and/or treatment of a KRAS G12D-mediated related disease.

In another aspect, the present invention provides the use of a compound of the present invention or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound or metabolite, or a pharmaceutical composition of the present invention, for the manufacture of a medicament for the prevention and/or treatment of KRAS G12D mediated related diseases.

In another aspect, the present invention provides a method for preventing and/or treating KRAS G12D-mediated related diseases, comprising administering to a subject in need thereof a prophylactically or therapeutically effective amount of a compound of the present invention, or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound or metabolite thereof, or a pharmaceutical composition of the present invention.

In another aspect, the invention provides methods of making the compounds of the invention.

Definition of

Unless defined otherwise below, all technical and scientific terms used herein are intended to have the same meaning as commonly understood by one of ordinary skill in the art. Reference to the techniques used herein is intended to refer to those techniques commonly understood in the art, including those variations of or alternatives to those techniques that would be apparent 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 "comprises," "comprising," "has," "containing," or "involving," and other variations thereof herein, are inclusive or open-ended and do not exclude additional unrecited elements or method steps.

The term "alkyl" as used herein 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, etc.), which is optionally substituted with one or more (such as 1 to 3) suitable substituents (e.g., halo, etc.).

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" as used herein _2-6 Alkenyl "means an alkenyl group having 2 to 6 carbon atoms and one, two or three (preferably one) carbon-carbon double bonds (e.g., vinyl, 1-propenyl, 2-butenyl, 3-butenyl, 2-pentenyl, 3-pentenyl, 4-pentenyl, 2-hexenyl, 3-hexenyl, 4-hexenyl, 5-hexenyl, 2-methyl-2-propeneYl, 4-methyl-3-pentenyl, etc.), which is optionally substituted with one or more (e.g., 1 to 3) suitable substituents (e.g., halo, etc.).

As used herein, the term "alkynyl" refers to a straight or branched aliphatic hydrocarbon group having one or more carbon-carbon triple bonds. For example, the term "C" as used herein _2-6 Alkynyl "refers to alkynyl groups having 2 to 6 carbon atoms and one, two or three (preferably one) carbon-carbon triple bonds (e.g., ethynyl, 1-propynyl, 2-butynyl, 3-butynyl, 2-pentynyl, 3-pentynyl, 4-pentynyl, 2-hexynyl, 3-hexynyl, 4-hexynyl, 5-hexynyl, etc.), optionally substituted with one or more (e.g., 1 to 3) suitable substituents (e.g., halogen, etc.).

As used herein, the term "cycloalkyl" refers to a saturated or partially unsaturated non-aromatic monocyclic or polycyclic (such as bicyclic) hydrocarbon ring (e.g., monocyclic, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, or bicyclic, including spiro, fused or bridged systems, such as bicyclo [1.1.1]Pentyl, bicyclo [2.2.1 ] s]Heptyl, bicyclo [3.2.1]Octyl or bicyclo [5.2.0]Nonyl, decalinyl, etc.), 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, etc.) hydrocarbon ring (e.g., cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl) having 3 to 6 ring carbon atoms, which is optionally substituted by one or more (such as 1 to 3) suitable substituents, for example, methyl-substituted cyclopropyl.

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 to 3) identical or different halogen atoms. For example, the term "C _1-6 Haloalkyl "means haloalkyl having 1 to 6 carbon atoms, e.g. -CF ₃ 、-C ₂ F ₅ 、-CHF ₂ 、-CH ₂ F、-CH ₂ CF ₃ 、-CH ₂ Cl or-CH ₂ CH ₂ CF ₃ And so on.

As used herein, the term "heterocycle" or "heterocyclyl" refers to a saturated or partially unsaturated non-aromatic monocyclic or polycyclic group, e.g., having 2,3, 4,5, 6,7,8, or 9 carbon atoms in the ring and one or more (e.g., 1,2,3, or 4) independently selected from N, O, or S (O) _t (wherein t is 0, 1 or 2), such as a 3-12 membered heterocyclic group, a 3-7 membered heterocyclic group, a 3-6 membered heterocyclic group, a 5-6 membered heterocyclic group and the like. Representative examples of heterocyclyl groups include, but are not limited to, oxiranyl, aziridinyl, azetidinyl, oxetanyl, tetrahydrofuranyl, pyrrolidinyl, hexahydro-1H-pyrrolinyl, hexahydropyrrolizinyl, pyrrolidinonyl, imidazolidinyl, pyrazolidinyl, tetrahydropyranyl, piperidinyl, morpholinyl, dithianyl, thiomorpholinyl, piperazinyl, and the like.

As used herein, the term "spiro" or "spirocycloalkyl" refers to a polycyclic group sharing one carbon atom (referred to as a spiro atom) between monocycloalkyl groups, which may contain one or more double bonds, but none of the rings have a completely conjugated pi-electron system. For example 8 to 11 membered. Spirocycloalkyl groups are classified as mono-spirocycloalkyl, di-spirocycloalkyl or multi-spirocycloalkyl according to the number of spiro atoms shared between rings.

As used herein, the term "spiroheterocycle" or "spiroheterocycloalkyl" refers to a polycyclic heterocyclic group that shares one atom (referred to as a spiro atom) between monocyclic rings, wherein one or more ring atoms is a heteroatom selected from nitrogen, oxygen, or S, and the remaining ring atoms are carbon. It may contain one or more double bonds, but no ring has a completely conjugated pi-electron system. For example 8 to 11 membered. Spiro heterocyclic groups are classified into a mono-spiro heterocyclic group, a di-spiro heterocyclic group, or a multi-spiro heterocyclic group according to the number of spiro atoms shared between rings.

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 Aromatic ring "means an aromatic group containing 6 to 10 carbon atomsGroups such as phenyl (ring) or naphthyl (ring). Aryl is optionally substituted with 1 or more (such as 1 to 3) suitable substituents (e.g. halogen, -OH, -CN, -NO) ₂ 、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, in particular containing 1 or 2 or 3 or 4 or 5 or 6 or 9 or 10 carbon atoms, and additionally may in each case be benzo-fused. For example, the heteroaryl or heteroaromatic ring may be selected from thienyl (ring), furyl (ring), pyrrolyl (ring), oxazolyl (ring), thiazolyl (ring), imidazolyl (ring), pyrazolyl (ring), isoxazolyl (ring), isothiazolyl (ring), oxadiazolyl (ring), triazolyl (ring), thiadiazolyl (ring), and the like, and benzo derivatives thereof; or pyridyl (ring), pyridazinyl (ring), pyrimidinyl (ring), pyrazinyl (ring), triazinyl (ring), etc., and benzo derivatives thereof.

The term "substituted" means that one or more (e.g., 1,2,3, or 4) hydrogens on the designated atom is replaced with a selection from the indicated group, provided that the designated atom's normal valency at the present time is not exceeded and the substitution results in a stable compound. Combinations of substituents and/or variables are permissible only if such combinations result in stable compounds.

If a substituent is described as "optionally substituted with \8230, the substituent may be (1) unsubstituted or (2) substituted. If a carbon of a substituent is described as being optionally substituted with one or more of a list of substituents, then one or more hydrogens on the carbon (to the extent of any hydrogens present) may be replaced individually and/or together with an independently selected substituent or not. If a nitrogen of a substituent is described as being optionally substituted with one or more of a list of substituents, then one or more hydrogens on the nitrogen (to the extent any hydrogens present) may each be replaced with an independently selected substituent or not.

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

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, under reasonable conditions.

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 of a substituent is shown through a bond connecting two atoms in a ring, then such substituent may be bonded to any ring atom in the substitutable ring.

The invention also includes all pharmaceutically acceptable isotopically-labeled compounds, which are identical to those of the present invention, except that one or more atoms are replaced by an atom having the same atomic number, but an atomic mass or mass number different from the atomic mass or mass number predominant in nature. Examples of isotopes suitable for inclusion into compounds of the invention include, but are not limited to, isotopes of hydrogen (e.g., hydrogen) ² H、 ³ H. Deuterium D, tritium T); isotopes of carbon (e.g. of ¹¹ C、 ¹³ C and ¹⁴ c) (ii) a Isotopes of chlorine (e.g. of chlorine) ³⁷ Cl); isotopes of fluorine (e.g. of fluorine) ¹⁸ F) (ii) a Isotopes of iodine (e.g. of iodine) ¹²³ I and ¹²⁵ i) (ii) a Isotopes of nitrogen (e.g. of ¹³ N and ¹⁵ n); isotopes of oxygen (e.g. of ¹⁵ O、 ¹⁷ O and ¹⁸ o); isotopes of phosphorus (e.g. of phosphorus) ³² P); and isotopes of sulfur (e.g. of ³⁵ S). Certain isotopically-labeled compounds of the present invention (e.g., those into which a radioactive isotope is incorporated) are useful in drug and/or substrate tissue distribution studies (e.g., assays). Radioisotope tritium (i.e. tritium ³ H) And carbon-14 (i.e. ¹⁴ C) Are particularly useful for this purpose because of their ease of incorporation and their ease of detection. With positron-emitting isotopes (e.g.) ¹¹ C、 ¹⁸ F、 ¹⁵ O and ¹³ n) can be used to examine substrate receptor occupancy in Positron Emission Tomography (PET) studies. Isotopically-labelled compounds of the present invention can be prepared bySimilar methods to those described in the accompanying schemes and/or examples and preparations are prepared by using appropriate isotopically labeled reagents in place of the non-labeled reagents previously employed. Pharmaceutically acceptable solvates of the invention include those in which the crystallization solvent may be isotopically substituted, e.g., D ₂ O, acetone-d ₆ Or DMSO-d ₆ 。

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

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

It will also be appreciated that certain compounds of the invention may be present in free form for use in therapy or, where appropriate, in the form of a pharmaceutically acceptable derivative thereof. In the present invention, pharmaceutically acceptable derivatives include, but are not limited to: pharmaceutically acceptable salts, solvates or metabolites, which upon administration to a patient in need thereof are capable of providing, directly or indirectly, a compound of the invention or a metabolite or residue thereof. Thus, when reference is made herein to "a compound of the invention", it is also intended to encompass the various derivative forms of the compound described above.

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. A review of suitable Salts is given in Stahl and Wermuth, handbook of Pharmaceutical Salts: properties, selection, and Use (Wiley-VCH, 2002). Methods for preparing pharmaceutically acceptable salts of the compounds of the present invention are known to those skilled in the art.

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

Those skilled in the art will appreciate that not all nitrogen-containing heterocycles are capable of forming N-oxides, since the available lone pair is required for oxidation of the nitrogen to the oxide; one skilled in the art will recognize nitrogen-containing heterocycles that are capable of forming N-oxides. Those skilled in the art will also recognize that tertiary amines are capable of forming N-oxides. Synthetic methods for preparing N-oxides of heterocycles and tertiary amines are well known to those skilled in the art and include oxidation of heterocycles and tertiary amines with peroxy acids such as peracetic and m-chloroperbenzoic acid (MCPBA), hydrogen peroxide, alkyl hydroperoxides such as t-butyl hydroperoxide, sodium perborate, and dioxiranes (dioxiranes) such as dimethyldioxirane. These methods for preparing N-oxides have been widely described and reviewed in the literature, see for example: T.L.Gilchrist, comprehensive Organic Synthesis, vol.7, pp 748-750; a.r.katitzky and a.j.boulton, eds., academic Press; and g.w.h.cheeseman and e.s.g.werstink, advances in Heterocyclic Chemistry, vol.22, pp 390-392, a.r.kattritzky and a.j.boulton, eds., academic Press.

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

The invention also encompasses compounds of the invention containing a protecting group. In any process for the preparation of the compounds of the present invention, it may be necessary and/or desirable to protect sensitive or reactive groups on any of the molecules concerned, thereby forming a chemically protected form of the compounds of the present invention. This can be achieved by conventional protecting Groups, as described, for example, in Protective Groups in Organic Chemistry, ed.j.f.w.mcmoie, plenum Press,1973; and T.W.Greene & P.G.M.Wuts, protective Groups in Organic Synthesis, john Wiley & Sons,1991, which are incorporated herein by reference. The protecting group may be removed at a suitable subsequent stage using methods known in the art.

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

Compound (I)

It is an object of the present invention to provide a compound of formula (I) or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound or metabolite thereof:

wherein,

ring A is 5-7 membered heterocycle, 5-6 membered heteroaromatic ring, C _5-7 Cycloalkyl, phenyl ring, 8-11 membered spiroheterocycle or C _8-11 Spirocycloalkyl, the A ring optionally substituted by one or more R ³ Substitution;

X ¹ selected from N and CR ⁴ ；

L ² Selected from the group consisting of covalent bonds, -O-, and-C (O) -;

R ¹ selected from hydrogen, C _1-6 Alkyl radical, C _3-10 Cycloalkyl and 3-12 membered heterocyclyl, each of which is optionally substituted with one or more R ⁶ Substitution;

R ³ selected from hydrogen, halogen, hydroxy, cyano, C _1-6 Alkyl radical, C _1-6 Haloalkyl, C _2-6 Alkenyl radical, C _2-6 Alkynyl, -O-C _1-6 Alkyl, -O-C _3-6 Cycloalkyl, -O-C _2-6 Alkenyl, -O-C _2-6 Alkynyl, C _3-10 Cycloalkyl and 3-12 membered heterocyclyl; or two R of the same carbon atom ³ And the carbon atoms to which they are both attached form a carbonyl group;

R ⁵ selected from hydrogen and C _1-6 An alkyl group;

R ⁷ Selected from hydrogen, halogen, hydroxy, cyano、C _1-6 Alkyl radical, C _2-6 Alkenyl radical, C _2-6 Alkynyl, -O-C _1-6 Alkyl, -O-C _3-6 Cycloalkyl radical, C _3-10 Cycloalkyl, 3-12 membered heterocyclyl and 5-10 membered heteroaryl, each of said alkyl, cycloalkyl, heterocyclyl or heteroaryl optionally substituted with one or more groups selected from hydrogen, halogen, hydroxy, cyano, C _1-6 Alkyl and C _1-6 Substituted with a substituent of haloalkyl; or,

R ³ and R ⁷ Together with adjacent atoms to form C _5-7 Cycloalkyl or 5-7 membered heterocycle;

R ⁸ selected from halogen, hydroxy, cyano, C _1-6 Alkyl, -C _1-6 alkylene-OH, -C _1-6 Alkyl-cyano and C _1-6 A haloalkyl group;

R ¹⁰ selected from hydrogen and C _1-6 An alkyl group;

R ¹¹ selected from hydrogen, C _1-6 Alkyl radical, C _1-6 Haloalkyl, C _3-10 Cycloalkyl, 3-12 membered heterocyclyl, each of which is optionally substituted with one or more substituents selected from hydrogen, halogen, hydroxy, cyano, C _1-6 Alkyl radical, C _1-6 Substituted with a haloalkyl;

R ¹² selected from hydrogen and C _1-6 An alkyl group;

n is 0, 1 or 2;

o, p, q, r and s are each independently selected from 0, 1,2 or 3, and o and r are not 0 at the same time, p and q are not 0 at the same time;

provided that when

Is->

Is->

-L ¹ -R ¹ Is composed of

When, -L ² -R ² Is not->

In some embodiments, the present invention provides a compound of formula (I), or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, or metabolite thereof, wherein,

X ¹ selected from N and CR ⁴ ；

L ² Selected from the group consisting of covalent bonds, -O-, and-C (O) -;

R ³ selected from hydrogen, halogen, hydroxy, cyano, C _1-6 Alkyl radical, C _1-6 Haloalkyl, C _2-6 Alkenyl radical, C _2-6 Alkynyl, -O-C _1-6 Alkyl, -O-C _3-6 Cycloalkyl, -O-C _2-6 Alkenyl, -O-C _2-6 Alkynyl, C _3-10 Cycloalkyl and 3-a 12 membered heterocyclyl group; or two R of the same carbon atom ³ And the carbon atoms to which they are both attached form a carbonyl group;

R ⁵ selected from hydrogen and C _1-6 An alkyl group;

R ⁷ Selected from hydrogen, halogen, hydroxy, cyano, C _1-6 Alkyl radical, C _2-6 Alkenyl radical, C _2-6 Alkynyl, -O-C _1-6 Alkyl, -O-C _3-6 Cycloalkyl radical, C _3-10 Cycloalkyl, 3-12 membered heterocyclyl and 5-10 membered heteroaryl, each of said alkyl, cycloalkyl, heterocyclyl or heteroaryl optionally substituted with one or more groups selected from hydrogen, halogen, hydroxy, cyano, C _1-6 Alkyl and C _1-6 Substituted with a haloalkyl; or,

R ⁹ selected from hydrogen, C _1-6 Alkyl radical, C _1-6 Haloalkyl, -C _1-6 alkylene-O-C _1-6 Alkyl radical, C _3-10 Cycloalkyl, 3-12 membered heterocyclyl, each of which is optionally substituted with one or more substituents selected from hydrogen, halogen, hydroxy, cyano, C _1-6 Alkyl radical, C _1-6 Substituted with a substituent of haloalkyl;

R ¹⁰ selected from hydrogen and C _1-6 An alkyl group;

R ¹² selected from hydrogen and C _1-6 An alkyl group;

n is 0, 1 or 2;

o, p, q, r and s are each independently selected from 0, 1,2 or 3, and o and r are not 0 at the same time and p and q are not 0 at the same time.

In some embodiments, R ⁴ Selected from hydrogen, halogen, cyano, C _1-4 Alkyl and C _1-4 A haloalkyl group.

In some embodiments, R ⁴ Selected from hydrogen, halogen, C _1-4 Alkyl and C _1-4 A haloalkyl group.

In some embodiments, R ⁴ Selected from hydrogen, halogen and cyano.

In some embodiments, R ⁴ Selected from hydrogen, F and cyano.

According to some embodiments of the invention, X ¹ Selected from N, CF, C-CN and CH.

In some embodiments, X ¹ Selected from N and CH.

In a preferred embodiment, X ¹ Is N.

According to some embodiments of the invention, L ² Selected from the group consisting of covalent bonds and-C (O) -. In the preferred embodimentIn scheme, L ² Is a covalent bond.

According to some embodiments of the invention, the compound of the invention has the structure of formula (II-A), formula (II-B), formula (II-C), formula (II-D), or formula (II-E):

wherein m is 0, 1 or 2, each radical L ¹ 、R ¹ 、R ² 、R ³ 、R ⁴ 、R ⁸ N, o, p, q, r and s are as defined above.

In some embodiments, L ¹ Selected from covalent bonds, -O-, -C (O) -, and-S-, -NH-and-N (C) _1-3 Alkyl) -;

R ¹ is selected from C _1-4 Alkyl radical, C _3-6 Cycloalkyl and 3-8 membered heterocyclyl, each of said alkyl, cycloalkyl and heterocyclyl being optionally substituted with one or more R ⁶ Substitution; each R ⁶ Each independently selected from-OR ⁹ 、-O-C(O)-N(R ⁹ )R ¹⁰ 、-N(R ⁹ )R ¹⁰ 、-N(R ¹⁰ )-C(O)-R ⁹ 、-N(R ¹⁰ )-C(O)-OR ⁹ 、-C(O)-N(R ⁹ )R ¹⁰ 、C _3-10 Cycloalkyl, 5-10 membered heteroaryl, and 3-12 membered heterocyclyl, each of which is optionally substituted with one or more substituents independently selected from the group consisting of: hydrogen, halogen, hydroxy, amino, cyano, C _1-6 Alkyl, -C _1-6 alkylene-R ¹¹ 、-C _1-6 alkylene-OR ¹¹ and-C _1-6 alkylene-NR ¹¹ R ¹² ；

R ⁹ Selected from hydrogen, C _1-4 Alkyl radical, C _1-4 Haloalkyl, -C _1-4 alkylene-O-C _1-4 Alkyl radical, C _3-6 Cycloalkyl, 3-8 membered heterocyclyl, each of which is optionally substituted with one or more substituents selected from hydrogen, halogen, hydroxy, cyano, C _1-6 Alkyl radical, C _1-6 Substituted with a haloalkyl;

R ¹⁰ selected from hydrogen and C _1-6 An alkyl group;

R ¹¹ selected from hydrogen, C _1-6 Alkyl radical, C _1-6 Haloalkyl, C _3-10 Cycloalkyl, 3-12 membered heterocyclyl, each of which is optionally substituted with one or more substituents selected from fluoro, chloro, bromo, iodo, hydroxy, cyano, C _1-6 Alkyl radical, C _1-6 Substituted with a haloalkyl;

R ¹² selected from hydrogen and C _1-6 An alkyl group;

R ⁷ selected from hydrogen, halogen, hydroxy, cyano, C _1-6 Alkyl radical, C _2-6 Alkenyl radical, C _2-6 Alkynyl, -O-C _1-6 Alkyl, -O-C _3-6 Cycloalkyl radical, C _3-10 Cycloalkyl, 3-12 membered heterocyclyl and 5-10 membered heteroaryl, each optionally substituted with one or more substituents selected from hydrogen, halogen, hydroxy, cyano, C _1-6 Alkyl and C _1-6 Substituted with a substituent of haloalkyl; or,

n is 0, 1 or 2;

o, p, q, r and s are each independently selected from 0, 1,2 or 3, and o and r are not simultaneously 0and p and q are not simultaneously 0.

According to some embodiments of the invention, L ¹ Selected from covalent bonds, -O-, -C (O) -, and-S-, -NH-and-N (C) _1-3 Alkyl) -. In a preferred embodiment, L ¹ Selected from-O-, -NH-and-N (C) _1-3 Alkyl) -. In a particularly preferred embodiment, L ¹ is-O-.

According to some embodiments of the invention, R ¹ Is selected from C _1-6 Alkyl radical, C _3-6 Cycloalkyl and 3-8 membered heterocyclyl, each of which is optionally substituted with one or more R ⁶ Substitution; each R ⁶ Each independently selected from hydrogen, halogen, cyano, -OR ⁹ 、-O-C(O)-N(R ⁹ )R ¹⁰ 、-O-C(O)-R ⁹ 、-N(R ⁹ )R ¹⁰ 、-N(R ¹⁰ )-C(O)-R ⁹ 、-N(R ¹⁰ )-C(O)-N(R ⁹ )R ¹⁰ 、-N(R ¹⁰ )-C(O)-OR ⁹ 、-C(O)-R ⁹ 、-C(O)-OR ¹⁰ 、-C(O)-N(R ⁹ )R ¹⁰ 、C _1-6 Alkyl radical, C _1-6 Haloalkyl, C _3-10 Cycloalkyl, C _6-10 Aryl, 5-10 membered heteroaryl, and 3-12 membered heterocyclyl, each of which is optionally substituted with one or more substituents independently selected from the group consisting of: hydrogen, halogen, hydroxy, amino, cyano, C _1-6 Alkyl radical, C _1-6 Haloalkyl, -C _1-6 alkylene-R ¹¹ 、-C _1-6 alkylene-OR ¹¹ and-C _1-6 alkylene-NR ¹¹ R ¹² 。

In a preferred embodiment, R ¹ Is selected from C _1-4 Alkyl radical, C _3-6 Cycloalkyl and 3-8 membered heterocyclyl, each of which is optionally substituted with one or more R ⁶ Substituted, each R ⁶ Each independently selected from-OR ⁹ 、-O-C(O)-N(R ⁹ )R ¹⁰ 、-N(R ⁹ )R ¹⁰ 、-N(R ¹⁰ )-C(O)-R ⁹ 、-N(R ¹⁰ )-C(O)-OR ⁹ 、-C(O)-N(R ⁹ )R ¹⁰ 、C _3-10 A cycloalkyl group, a,5-10 membered heteroaryl and 3-12 membered heterocyclyl, each of which is optionally substituted with one or more substituents independently selected from the group consisting of: hydrogen, halogen, hydroxy, amino, cyano, C _1-6 Alkyl, -C _1-6 alkylene-R ¹¹ 、-C _1-6 alkylene-OR ¹¹ and-C _1-6 alkylene-NR ¹¹ R ¹² ；

R ⁹ Selected from hydrogen, C _1-4 Alkyl radical, C _1-4 Haloalkyl, -C _1-4 alkylene-O-C _1-4 Alkyl radical, C _3-6 Cycloalkyl, 3-8 membered heterocyclyl, each of which is optionally substituted with one or more substituents selected from hydrogen, halogen, hydroxy, cyano, C _1-6 Alkyl radical, C _1-6 Substituted with a substituent of haloalkyl;

R ¹⁰ selected from hydrogen and C _1-6 An alkyl group;

R ¹¹ selected from hydrogen, C _1-6 Alkyl radical, C _1-6 Haloalkyl and 3-12 membered heterocyclyl;

R ¹² selected from hydrogen and C _1-6 An alkyl group.

In some embodiments, R ¹ Is C _1-4 An alkyl group optionally substituted with one or more R ⁶ Substituted, each R ⁶ Each independently selected from C _3-10 Cycloalkyl, 5-10 membered heteroaryl, and 3-12 membered heterocyclyl, each of which is optionally substituted with one or more substituents independently selected from the group consisting of: halogen, C _1-6 Alkyl, -C _1-6 alkylene-R ¹¹ and-C _1-6 alkylene-NR ¹¹ R ¹² ；

R ¹¹ Selected from hydrogen, C _1-6 Alkyl and 3-12 membered heterocyclyl;

R ¹² is C _1-6 An alkyl group.

In some embodiments, R ¹ Is C _1-4 Alkyl optionally substituted with one or more R ⁶ Substituted, each R ⁶ Each independently selected from C _3-10 Cycloalkyl, 5-10 membered heteroaryl and 3-12 membered heterocyclyl, said cycloalkyl, heteroarylAnd heterocyclyl are each optionally substituted with one or more substituents independently selected from the group consisting of: c _1-6 Alkyl, -C _1-6 alkylene-R ¹¹ and-C _1-6 alkylene-NR ¹¹ R ¹² ；

R ¹¹ Selected from hydrogen, C _1-6 Alkyl and 3-12 membered heterocyclyl;

R ¹² is C _1-6 An alkyl group.

In some embodiments, R ¹ Is optionally substituted by one or more R ⁶ Substituted C _1-4 Alkyl radical, each R ⁶ Each independently selected from: c _3-6 Cycloalkyl, 5-6 membered heteroaryl, and 5-8 membered heterocyclyl; the cycloalkyl, heteroaryl and heterocyclyl are each optionally substituted with one or more substituents independently selected from the group consisting of: halogen, C _1-6 Alkyl, -C _1-6 alkylene-R ¹¹ and-C _1-6 alkylene-NR ¹¹ R ¹² ；R ¹¹ Is selected from C _1-6 Alkyl and 3-12 membered heterocyclyl; r ¹² Is C _1-6 An alkyl group.

In some embodiments, R ¹ Is optionally substituted by one or more R ⁶ Substituted C _1-4 Alkyl radical, each R ⁶ Each independently selected from: 5-6 membered heteroaryl and 5-8 membered heterocyclyl; each of said heteroaryl and heterocyclyl is optionally substituted with one or more substituents independently selected from the group consisting of: halogen and C _1-6 An alkyl group.

In some embodiments, R ¹ Is optionally substituted by one or more R ⁶ Substituted C _1-4 Alkyl radical, each R ⁶ Each independently selected from: cyclopropyl, pyrrolidinyl, hexahydropyrazinyl and imidazolyl, each optionally substituted with one or more substituents independently selected from the group consisting of: halogen, C _1-6 Alkyl, -C _1-6 alkylene-R ¹¹ and-C _1-6 alkylene-NR ¹¹ R ¹² ；R ¹¹ Is selected from C _1-6 Alkyl and 3-12 membered heterocyclyl; r ¹² Is C _1-6 An alkyl group.

In part realIn the embodiment, R ¹ Is optionally substituted by one or more R ⁶ Substituted C _1-4 Alkyl radical, each R ⁶ Each independently selected from: cyclopropyl, pyrrolidinyl, hexahydropyrazinyl and imidazolyl, each optionally substituted with one or more substituents independently selected from the group consisting of: halogen, C _1-6 Alkyl, -C _1-6 Alkylene-morpholinyl and-C _1-6 alkylene-N (C) _1-6 Alkyl radical) ₂ 。

In some embodiments, R ¹ Is selected from

Wherein the wave line pick>

Indicates the point of attachment of the group to the rest of the molecule.

In some embodiments, R ¹ Is selected from

Wherein the wave line pick>

Indicates the point of attachment of the group to the rest of the molecule.

In some embodiments, R ¹ Is selected from

Wherein the wave line pick>

Indicates the point of attachment of the group to the rest of the molecule.

According to some embodiments of the invention, R ² Is selected from C _6-10 Aryl and 5-10 membered heteroaryl, each of which is optionally substituted with oneA plurality of R ⁷ Substitution; each R ⁷ Each independently selected from hydrogen, halogen, hydroxy, cyano, C _1-3 Alkyl radical, C _2-4 Alkenyl radical, C _2-4 Alkynyl, -O-C _1-3 Alkyl, -O-C _3-6 Cycloalkyl, C _3-6 Cycloalkyl and 3-8 membered heterocyclyl, each of said alkyl, cycloalkyl and heterocyclyl being optionally substituted with one or more substituents selected from hydrogen, halogen, hydroxy, cyano, C _1-3 Alkyl and C _1-3 Substituted with a haloalkyl.

In a preferred embodiment, R ² Is selected from C _6-10 Aryl and 5-10 membered heteroaryl, each optionally substituted with one or more R ⁷ Substituted, each R ⁷ Each independently selected from hydrogen, halogen, hydroxy, C _1-3 Alkyl radical, C _2-4 Alkynyl, -O-C _1-3 Alkyl and C _3-6 Cycloalkyl, each of which is optionally substituted with one or more of hydrogen and halogen.

In some embodiments, R ² Is selected from C _6-10 Aryl and 5-10 membered heteroaryl, each optionally substituted with one or more substituents independently selected from halogen, hydroxy, C _1-3 Alkyl and C _2-4 And substituent of alkynyl.

In some embodiments, R ² Is selected from C _6-10 Aryl and 5-10 membered heteroaryl, each optionally substituted with one or more substituents independently selected from F, cl, br, hydroxy, methyl, ethyl, n-propyl, isopropyl, ethynyl and propynyl.

In some embodiments, R ² Is naphthalene, optionally substituted with one or more substituents independently selected from the group consisting of F, cl, br, hydroxy, ethyl and ethynyl.

In some embodiments, R ² Is selected from C _6-10 Aryl and 5-10 membered heteroaryl, each of which is optionally substituted with one or more hydroxyl groups.

In some embodiments, R ² Is selected from

In some embodiments, each R is ³ Each independently selected from hydrogen, halogen, hydroxy, cyano, C _1-4 Alkyl radical, C _1-4 Haloalkyl, C _2-4 Alkenyl radical, C _2-4 Alkynyl, -O-C _1-4 Alkyl, -O-C _3-6 Cycloalkyl, -O-C _2-4 Alkenyl, -O-C _2-4 Alkynyl, C _3-6 Cycloalkyl and 3-8 membered heterocyclyl; or two R of the same carbon atom ³ And the carbon atom to which they are attached together form a carbonyl group.

In some embodiments, each R is ³ Each independently selected from hydrogen, halogen, hydroxy, C _1-4 Alkyl radical, C _1-4 Haloalkyl, -O-C _1-4 Alkyl, -O-C _3-6 Cycloalkyl radical, C _3-6 Cycloalkyl and 3-8 membered heterocyclyl.

In some embodiments, each R is ³ Each independently selected from hydrogen and halogen.

In some embodiments, each R is ³ Each independently is halogen; preferably, each R ³ Each independently selected from F and Cl.

In some embodiments, R ⁹ Selected from hydrogen, C _1-4 Alkyl radical, C _1-4 Haloalkyl, -C _1-4 alkylene-O-C _1-4 Alkyl radical, C _3-6 Cycloalkyl, 3-8 membered heterocyclyl, said cycloalkyl or heterocyclyl being optionally substituted by one or more substituents selected from hydrogen, halogen, hydroxy, cyano, C _1-6 Alkyl radical, C _1-6 Substituted with a haloalkyl.

In some embodiments, R ⁹ Selected from hydrogen, C _1-4 Alkyl and C _1-4 A haloalkyl group.

In some embodiments, R ¹¹ Selected from hydrogen, C _1-6 Alkyl radical, C _1-6 Haloalkyl, and 3-12 membered heterocyclyl.

In some embodiments, R ¹¹ Selected from hydrogen, C _1-6 Alkyl and 3-12 membered heterocyclyl.

In some embodiments, R ¹¹ Is selected from C _1-6 Alkyl and 3-12 membered heterocyclyl.

In some embodiments, R ¹¹ Selected from the group consisting of hydrogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, pentyl and morpholinyl.

In some embodiments, R ¹¹ Selected from methyl and morpholinyl.

In some embodiments, R ¹² Selected from the group consisting of hydrogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl and pentyl.

In some embodiments, R ¹² Is methyl.

In some embodiments, o, p are both 0, q, r are both 1, and s is 1,2, or 3.

In some embodiments, o, p are both 0, q, r are both 1, and s is 2.

In some embodiments, o, p are both 0, and q, r, s are all 1.

In some embodiments, o, p are both 0, q, r are both 1, and s is 3.

In some of the embodiments of the present invention,

selected from the group consisting of>

According to some embodiments of the invention, the compounds of the invention have the structure of formula (II-A-1), (II-A-2), (II-A-3), formula (II-B-1), formula (II-C-1), formula (II-D-1), or formula (II-E-1):

wherein each group L, R ¹ 、R ² 、R ³ 、R ⁴ And m is as defined above.

The present invention encompasses compounds of formula (I) obtained by any combination of the above preferred groups.

According to some embodiments of the invention, the compound of the invention is selected from:

preparation method

It is another object of the present invention to provide a process for preparing the compounds of the present invention. For example, the present invention provides a process for preparing a compound of formula (II-a) comprising the steps of:

wherein L is ¹ 、R ¹ 、R ² 、R ³ 、R ⁸ M, n, o, p, q, r and s are as defined above;

LG ¹ 、LG ² 、LG ³ and X represents a leaving group including, but not limited to, a halogen atom, trifluoromethanesulfonate, methylthio, methylsulfinyl, methylsulfonyl, or the like;

PG ¹ 、PG ² and PG ³ Denotes a protecting group including, but not limited to, benzyloxycarbonyl (Cbz), t-butyloxycarbonyl (Boc), methylA phenyl group or benzyl group, etc.;

(1) Reacting compound II-A1-1 with compound II-A1-2 to give compound II-A1-3;

the reaction is preferably carried out in a suitable organic solvent. The organic solvent may be selected from the group consisting of methanol, toluene, tetrahydrofuran, 1, 4-dioxane, and any combination thereof, preferably methanol. The reaction is preferably carried out in the presence of a suitable organic or inorganic base which may be selected from diisopropylethylamine, triethylamine, sodium methoxide, sodium tert-butoxide, preferably sodium methoxide. The reaction is carried out at a suitable temperature, preferably from 0 to 80 ℃. The reaction is carried out for a suitable time, for example 2 to 24 hours.

(2) Carrying out coupling reaction on the compound II-A1-3 and the compound II-A-4 to obtain a compound II-A-5;

the coupling reaction is preferably carried out in the presence of a metal catalyst, a ligand and a base. Preferably, the metal catalyst is a palladium metal catalyst, such as tris (dibenzylideneacetone) dipalladium, [1,1 '-bis (diphenylphosphino) ferrocene ] palladium dichloride dichloromethane complex, tetrakis (triphenylphosphine) palladium, palladium acetate, preferably [1,1' -bis (diphenylphosphino) ferrocene ] palladium dichloride dichloromethane complex. Preferably, the ligand is a phosphorus ligand, such as 4, 5-bis-diphenylphosphine-9, 9-dimethylxanthene, 2-dicyclohexylphosphine-2 ',6' -diisopropoxy-1, 1 '-biphenyl, 2-dicyclohexylphosphine-2', 4',6' -triisopropylbiphenyl, 2-dicyclohexylphosphine-2 ',6' -dimethoxy-biphenyl, preferably 4, 5-bis-diphenylphosphine-9, 9-dimethylxanthene. The base is an organic or inorganic base, such as diisopropylethylamine, triethylamine, sodium tert-butoxide, potassium carbonate, cesium carbonate, sodium carbonate, preferably cesium carbonate. The reaction is preferably carried out in a suitable solvent which may be selected from the group consisting of N, N-dimethylformamide, N-methylpyrrolidone, toluene, ethanol, ethylene glycol dimethyl ether, water, 1, 4-dioxane and any combination thereof, preferably 1, 4-dioxane. The reaction is carried out at a suitable temperature, preferably from 50 to 120 ℃. The reaction is carried out for a suitable time, for example 1 to 16 hours.

(3) Subjecting compound II-A-5 to a deprotection reaction to give compound II-A-6;

the deprotection reaction is preferably carried out in the presence of a metal catalyst. Preferably, the metal catalyst is a palladium metal catalyst or a platinum metal catalyst, such as palladium on carbon, palladium hydroxide, platinum dioxide, preferably palladium on carbon. The reduction reaction is preferably carried out in the presence of hydrogen. The reaction is preferably carried out in a suitable organic solvent which may be selected from the group consisting of methanol, ethanol, isopropanol, ethyl acetate, tetrahydrofuran and any combination thereof, preferably methanol. The reaction is carried out at a suitable temperature, preferably 20-60 ℃. The reaction is carried out for a suitable time, for example 2 to 16 hours.

(4) Carrying out coupling reaction on the compound II-A-6 and the compound II-A-7 to obtain a compound II-A-8;

the coupling reaction is preferably carried out in the presence of a metal catalyst, a ligand and a base. Preferably, the metal catalyst is a palladium metal catalyst, such as tris (dibenzylideneacetone) dipalladium, [1,1' -bis (diphenylphosphino) ferrocene ] dichloropalladium, tetrakis (triphenylphosphine) palladium, palladium acetate, preferably tris (dibenzylideneacetone) dipalladium. Preferably, the ligand is a phosphorus ligand, such as 4, 5-bis diphenylphosphine-9, 9-dimethylxanthene, 2-dicyclohexylphosphine-2 ',6' -diisopropoxy-1, 1' -biphenyl, 2-dicyclohexylphosphine-2 ',4',6' -triisopropylbiphenyl, 2-dicyclohexylphosphine-2 ',6' -dimethoxy-biphenyl, preferably 2-dicyclohexylphosphine-2 ',6' -diisopropoxy-1, 1' -biphenyl. The base is an organic or inorganic base, such as sodium tert-butoxide, potassium carbonate, cesium carbonate, sodium carbonate, preferably cesium carbonate. The reaction is preferably carried out in a suitable solvent which may be selected from the group consisting of N, N-dimethylformamide, N-methylpyrrolidone, toluene, ethanol, ethylene glycol dimethyl ether, water, 1, 4-dioxane and any combination thereof, preferably 1, 4-dioxane. The reaction is carried out at a suitable temperature, preferably from 50 to 120 ℃. The reaction is carried out for a suitable time, for example 2 to 16 hours.

(5) Subjecting compound II-A-8 to a deprotection reaction to give compound II-A-9;

the deprotection reaction is preferably carried out in a suitable organic solvent. The organic solvent may be selected from alcoholic protic solvents, tetrahydrofuran, dichloromethane, toluene, N-dimethylformamide and any combination thereof, preferably N, N-dimethylformamide. The deprotection reaction is preferably carried out in the presence of a suitable deprotection reagent, such as boron tribromide, trimethyliodosilane, acetic acid hydrobromide, palladium on carbon, palladium hydroxide, sodium ethanethiol, preferably sodium ethanethiol. The deprotection reaction is carried out at a suitable temperature, which is 0-100 ℃. The reaction is carried out for a suitable time, for example 1 to 12 hours.

(6) Reacting compound II-A-9 to obtain compound II-A-10;

the reaction is preferably carried out in the presence of a suitable halo-or pseudohalo-forming reagent which is trifluoromethanesulfonic anhydride, phosphorus oxychloride or phosphorus tribromooxybromide, preferably trifluoromethanesulfonic anhydride. The reaction is preferably carried out in a suitable organic solvent. The organic solvent may be selected from dichloromethane, toluene, tetrahydrofuran and any combination thereof, preferably dichloromethane. The reaction is preferably carried out in the presence of a suitable organic or inorganic base, such as diisopropylethylamine, triethylamine, N-dimethylaniline, preferably diisopropylethylamine. The reaction is carried out at a suitable temperature, preferably from-70 ℃ to 100 ℃. The reaction is carried out for a suitable time, for example 1 to 16 hours.

(7) Subjecting compound II-A-10 to a coupling reaction with compound II-A-11 to obtain compound II-A-12;

the coupling reaction is preferably carried out in the presence of a metal catalyst and a base. Preferably, the metal catalyst is a palladium metal catalyst, such as tris (dibenzylideneacetone) dipalladium, [1,1 '-bis (diphenylphosphino) ferrocene ] palladium dichloride dichloromethane complex, tetrakis (triphenylphosphine) palladium, palladium acetate, preferably [1,1' -bis (diphenylphosphino) ferrocene ] palladium dichloride dichloromethane complex. The base is an organic or inorganic base such as diisopropylethylamine, triethylamine, sodium tert-butoxide, potassium carbonate, cesium carbonate, sodium carbonate, preferably sodium carbonate. The reaction is preferably carried out in a suitable solvent which may be selected from the group consisting of N, N-dimethylformamide, N-methylpyrrolidone, toluene, ethanol, ethylene glycol dimethyl ether, water, 1, 4-dioxane and any combination thereof, preferably a combination of 1, 4-dioxane and water. The reaction is carried out at a suitable temperature, preferably from 50 to 120 ℃. The reaction is carried out for a suitable time, for example 2 to 16 hours.

(8) Subjecting the compound II-A-12 to a reduction reaction to obtain a compound II-A-13;

the reduction reaction is preferably carried out in the presence of a metal catalyst. Preferably, the metal catalyst is a palladium metal catalyst or a platinum metal catalyst, such as palladium on carbon, palladium hydroxide, platinum dioxide, preferably palladium on carbon. The reduction reaction is preferably carried out in the presence of hydrogen. The reaction is preferably carried out in a suitable organic solvent which may be selected from the group consisting of methanol, ethanol, isopropanol, ethyl acetate, tetrahydrofuran and any combination thereof, preferably methanol. The reaction is carried out at a suitable temperature, preferably from 20 to 60 ℃. The reaction is carried out for a suitable time, for example 2 to 16 hours.

(9) Subjecting the compound II-A-13 to a deprotection reaction to obtain the compound II-A.

The deprotection reaction is preferably carried out in a suitable organic solvent. The organic solvent may be selected from the group consisting of alcoholic protic solvents, tetrahydrofuran, dichloromethane, N-dimethylformamide and any combination thereof, preferably dichloromethane. The reaction is preferably carried out under suitable deprotection reagents. The deprotecting reagent may be selected from the group consisting of trimethylsilyl iodide, hydrobromic acid in acetic acid, and trifluoroacetic acid. Preferably a solution of hydrobromic acid in acetic acid. The deprotection reaction is carried out at a suitable temperature, preferably 0-100 ℃. The reaction is carried out for a suitable time, for example from 0.5 to 12 hours.

The present invention provides a second process for preparing a compound of formula (II-A) comprising the steps of:

wherein L is ¹ 、R ¹ 、R ² 、R ³ 、R ⁸ M, n, o, p, q, r and s are asAs defined above;

LG ¹ 、LG ² and X represents a leaving group including, but not limited to, a halogen atom, trifluoromethanesulfonate, methylthio, methylsulfinyl, methylsulfonyl, and the like;

PG ¹ and PG ³ Represents a protecting group including, but not limited to, benzyloxycarbonyl (Cbz), tert-butoxycarbonyl (Boc), benzyl, and the like;

(1) Subjecting compound II-A1-1 to a coupling reaction with compound II-A-11 to obtain compound IIa-A-2;

the coupling reaction is preferably carried out in the presence of a metal catalyst and a base. Preferably, the metal catalyst is a palladium metal catalyst, such as tris (dibenzylideneacetone) dipalladium, [1,1 '-bis (diphenylphosphino) ferrocene ] palladium dichloride dichloromethane complex, tetrakis (triphenylphosphine) palladium, palladium acetate, preferably [1,1' -bis (diphenylphosphino) ferrocene ] palladium dichloride dichloromethane complex. The base is an organic or inorganic base, such as N, N-diisopropylethylamine, triethylamine, sodium tert-butoxide, potassium carbonate, cesium carbonate, potassium acetate, sodium carbonate, preferably potassium acetate. The reaction is preferably carried out in a suitable solvent which may be selected from the group consisting of N, N-dimethylformamide, N-methylpyrrolidone, toluene, ethanol, ethylene glycol dimethyl ether, 1, 4-dioxane, water and any combination thereof, preferably a combination of 1, 4-dioxane and water. The reaction is carried out at a suitable temperature, preferably from 50 to 120 ℃. The reaction is carried out for a suitable time, for example 2 to 16 hours.

(2) Subjecting compound IIa-A-2 to a coupling reaction with compound II-A-4 to give compound IIa-A-3;

the coupling reaction is preferably carried out in the presence of a metal catalyst and a base. Preferably, the metal catalyst is a palladium metal catalyst, such as tris (dibenzylideneacetone) dipalladium, methanesulfonic acid (2-dicyclohexylphosphino-2 ',6' -diisopropoxy-1, 1' -biphenyl) (2-amino-1, 1' -biphenyl-2-yl) palladium (II), [1,1' -bis (diphenylphosphino) ferrocene ] dichloropalladium, tetrakis (triphenylphosphine) palladium, palladium acetate, preferably methanesulfonic acid (2-dicyclohexylphosphino-2 ',6' -diisopropoxy-1, 1' -biphenyl) (2-amino-1, 1' -biphenyl-2-yl) palladium (II). The base is an organic or inorganic base such as diisopropylethylamine, triethylamine, sodium tert-butoxide, potassium carbonate, cesium carbonate, sodium carbonate, preferably cesium carbonate. The reaction is preferably carried out in a suitable solvent which may be selected from the group consisting of N, N-dimethylformamide, N-methylpyrrolidone, toluene, ethanol, ethylene glycol dimethyl ether, 1, 4-dioxane, water and any combination thereof, preferably toluene. The reaction is carried out at a suitable temperature, preferably from 50 to 120 ℃. The reaction is carried out for a suitable time, for example 2 to 16 hours.

(3) Subjecting compound IIa-A-3 to a deprotection reduction reaction to obtain compound IIa-A-4;

the reaction is preferably carried out in the presence of a metal catalyst. Preferably, the metal catalyst is a palladium metal catalyst or a platinum metal catalyst, such as palladium on carbon, palladium hydroxide, platinum dioxide, preferably palladium on carbon. The reaction is preferably carried out in the presence of hydrogen. The reaction is preferably carried out in a suitable organic solvent which may be selected from the group consisting of ammonia methanol, ethanol, isopropanol, ethyl acetate, tetrahydrofuran and any combination thereof, preferably ammonia methanol. The reaction is preferably carried out at a suitable temperature, which is in the range of from 20 to 60 ℃. The reaction is carried out for a suitable time, for example 2 to 16 hours.

(4) Subjecting compound IIa-A-4 to a coupling reaction with compound II-A-7 to obtain compound II-A-13;

the coupling reaction is preferably carried out in the presence of a metal catalyst, a ligand and a base. Preferably, the metal catalyst is a palladium metal catalyst, such as tris (dibenzylideneacetone) dipalladium, [1,1' -bis (diphenylphosphino) ferrocene ] dichloropalladium, tetrakis (triphenylphosphine) palladium, palladium acetate, preferably tris (dibenzylideneacetone) dipalladium. The ligands are phosphorus ligands, for example 4, 5-bisdiphenylphosphine-9, 9-dimethylxanthene, 2-dicyclohexylphosphine-2 ',6' -diisopropoxy-1, 1' -biphenyl, 2-dicyclohexylphosphine-2 ',4',6' -triisopropylbiphenyl, 2-dicyclohexylphosphine-2 ',6' -dimethoxy-biphenyl, preferably 2-dicyclohexylphosphine-2 ',6' -diisopropoxy-1, 1' -biphenyl. The base is an organic or inorganic base, such as sodium tert-butoxide, potassium carbonate, cesium carbonate, sodium carbonate, preferably cesium carbonate. The reaction is preferably carried out in a suitable solvent which may be selected from the group consisting of N, N-dimethylformamide, N-methylpyrrolidone, toluene, ethanol, ethylene glycol dimethyl ether, 1, 4-dioxane, water and any combination thereof, preferably 1, 4-dioxane. The reaction is carried out at a suitable temperature, preferably from 50 to 120 ℃. The reaction is carried out for a suitable time, for example 2 to 16 hours.

(5) Subjecting compound II-A-13 to a deprotection reaction to give compound II-A;

the deprotection reaction is preferably carried out in a suitable organic solvent. The organic solvent may be selected from the group consisting of alcoholic protic solvents, tetrahydrofuran, dichloromethane, N-dimethylformamide and any combination thereof, preferably dichloromethane. The reaction is preferably carried out under suitable deprotection reagents. The deprotecting agent may be selected from the group consisting of trimethylsilyl iodide, hydrobromic acid in acetic acid, and trifluoroacetic acid. Trifluoroacetic acid is preferred. The deprotection reaction is carried out at a suitable temperature, preferably 0-80 ℃. The reaction is carried out for a suitable time, for example 0.5 to 12 hours.

The present invention provides a process for preparing a compound of formula (II-C) comprising the steps of:

LG ^1c 、LG ^2c 、LG ^3c 、LG ^4c and LG ^5c Each independently represents a leaving group such as halogen, triflate, thiomethyl, methyl sulfoxide, methyl sulfone, boronic acid, boronic ester, tributyltin, methoxy or ethoxy;

PG ^1c and PG ^2c Represents a protecting group including, but not limited to, benzyloxycarbonyl (Cbz), tert-butoxycarbonyl (Boc), methyl or benzyl, etc.;

(1) Reacting compound II-C1-1 with compound II-C-2 to give compound II-C-3;

(2) Reacting compound II-C-3 with compound II-A-4 to obtain compound II-C-4;

the reaction is preferably carried out in a suitable organic solvent and in the presence of a suitable organic base. The organic solvent may be selected from acetonitrile, N-dimethylformamide, dimethylsulfoxide, N-dimethylacetamide, tetrahydrofuran, 1, 4-dioxane, and any combination thereof, preferably 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 ℃ to 100 ℃ for 2 to 48 hours.

(3) Subjecting compound II-C-4 to a deprotection reaction to give compound II-C-5;

the deprotection reaction is preferably carried out in a suitable organic solvent. The organic solvent may be selected from the group consisting of alcoholic protic solvents, tetrahydrofuran, dichloromethane, toluene, acetonitrile and any combination thereof, preferably acetonitrile. The deprotection reaction is preferably carried out in the presence of a suitable deprotecting reagent, such as boron tribromide, trimethyliodosilane, acetic acid hydrobromide, palladium on carbon, palladium hydroxide, sodium ethanethiol, preferably trimethyliodosilane. The deprotection reaction is carried out at a suitable temperature, which is 0-100 ℃. The reaction is carried out for a suitable time, for example 1 to 12 hours.

(4) Reacting compound II-C-5 to obtain compound II-C-6;

the reaction is preferably carried out in the presence of a suitable halo-or pseudohalo-forming reagent which is trifluoromethanesulfonic anhydride, thionyl chloride, phosphorus oxychloride or phosphorus oxybromide, preferably thionyl chloride. The reaction is preferably carried out in the presence of a suitable catalyst which is N, N-dimethylaniline or N, N-dimethylformamide, preferably N, N-dimethylformamide. The reaction is carried out at a suitable temperature, preferably from-70 ℃ to 100 ℃. The reaction is carried out for a suitable time, for example 1 to 16 hours.

(5) Subjecting compound II-C-6 to a coupling reaction with compound II-C-7 to obtain compound II-C-8;

the coupling reaction is preferably carried out in the presence of a metal catalyst and a base. Preferably, the metal catalyst is a palladium metal catalyst, such as tris (dibenzylideneacetone) dipalladium, [1,1 '-bis (diphenylphosphino) ferrocene ] dichloropalladium dichloromethane complex, tetrakis (triphenylphosphine) palladium, palladium acetate, preferably [1,1' -bis (diphenylphosphino) ferrocene ] dichloropalladium dichloromethane complex. The base is an organic or inorganic base, such as N, N-diisopropylethylamine, triethylamine, sodium tert-butoxide, potassium carbonate, cesium carbonate, potassium acetate, sodium carbonate, preferably potassium acetate. The reaction is preferably carried out in a suitable solvent which may be selected from the group consisting of N, N-dimethylformamide, N-methylpyrrolidone, toluene, ethanol, ethylene glycol dimethyl ether, 1, 4-dioxane, water and any combination thereof, preferably a combination of 1, 4-dioxane and water. The reaction is carried out at a suitable temperature, preferably from 50 to 120 ℃. The reaction is carried out for a suitable time, for example 2 to 16 hours.

(6) Subjecting the compound II-C-8 to a reduction reaction to obtain a compound II-C-9;

the reaction is preferably carried out in the presence of a catalyst. Preferably, the catalyst is palladium on carbon, palladium hydroxide, nickel chloride or cobalt chloride hexahydrate, preferably cobalt chloride hexahydrate. The reaction is preferably carried out in the presence of a suitable reducing agent which is lithium aluminium hydride, sodium borohydride or hydrogen, preferably sodium borohydride. The reaction is preferably carried out in a suitable organic solvent which may be selected from the group consisting of methanol, ethanol, isopropanol, ethyl acetate, tetrahydrofuran and any combination thereof, preferably methanol. The reaction is preferably carried out at a suitable temperature, which is in the range of from 0 to 60 ℃. The reaction is carried out for a suitable time, for example 0.5 to 16 hours.

(7) Subjecting compound II-C-9 to a coupling reaction with compound II-C-10 to obtain compound II-C-11;

the coupling reaction 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 ] dichloropalladium, [ n-butylbis (1-adamantyl) phosphine ] (2-amino-1, 1' -biphenyl-2-yl) palladium (II), tetratriphenylphosphine palladium and palladium acetate, preferably [ n-butylbis (1-adamantyl) phosphine ] (2-amino-1, 1' -biphenyl-2-yl) palladium (II) methanesulfonate. The base may be an organic or inorganic base, for example selected from sodium tert-butoxide, potassium carbonate, potassium phosphate, cesium carbonate and sodium carbonate, preferably potassium phosphate. The solvent may be selected from the group consisting of N, N-dimethylformamide, N-methylpyrrolidone, toluene, ethanol, ethylene glycol dimethyl ether, water, 1, 4-dioxane, and any combination thereof, preferably a combination of 1, 4-dioxane and water. The reaction is preferably carried out at a temperature of 50 to 120 ℃ for 2 to 16 hours.

(8) Subjecting the compound II-C-11 to a deprotection reaction to obtain the compound II-C.

The deprotection reaction is preferably carried out in a suitable organic solvent. The organic solvent may be selected from the group consisting of alcoholic protic solvents, tetrahydrofuran, dichloromethane, N-dimethylformamide and any combination thereof, preferably dichloromethane. The reaction is preferably carried out under suitable deprotection reagents. The deprotecting agent may be selected from the group consisting of trimethylsilyl iodide, hydrobromic acid in acetic acid, and trifluoroacetic acid. Trifluoroacetic acid is preferred. The deprotection reaction is carried out at a suitable temperature, preferably 0-100 ℃. The reaction is carried out for a suitable time, for example from 0.5 to 12 hours.

Specific conditions for the above-mentioned reaction steps are known in the art, and the present invention is not particularly limited thereto. According to the teaching of the present invention and the common knowledge in the field, the skilled person can make selective substitutions on each substituent in the general formula to prepare different compounds, and the alternatives and substitutions are all within the protection scope of the present invention.

Pharmaceutical composition and kit

It is another object of the present invention to provide a pharmaceutical composition comprising a prophylactically or therapeutically effective amount of a compound of the present invention, or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound or metabolite thereof, and one or more pharmaceutically acceptable carriers.

It is another object of the present invention to provide a kit comprising a compound of the present invention or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound or metabolite thereof, or a pharmaceutical composition of the present invention, and optionally packaging and/or instructions.

By "pharmaceutically acceptable carrier" in the context of the present invention is meant a diluent, adjuvant, excipient, or vehicle that is administered together with a therapeutic agent and which is, within the scope of sound medical judgment, suitable for contact with the tissues of humans and/or other animals without excessive toxicity, irritation, allergic response, or other problem or complication 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.

The pharmaceutical composition may, for example, be in the form of a solid formulation, a semi-solid formulation, a liquid formulation, or a gaseous formulation, among others.

The pharmaceutical compositions of the present invention may act systemically and/or locally. For this purpose, they can be administered by a suitable route, for example by injection or transdermal administration; either orally or by inhalation.

The compound of the invention may be present in the pharmaceutical composition in an amount or amount of about 0.001mg to about 1000mg.

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

Methods of treatment and uses

It is another object of the present invention to provide a compound of the present invention, or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound or metabolite thereof, or a pharmaceutical composition of the present invention, for use in the inhibition of KRAS G12D.

It is another object of the present invention to provide a compound of the present invention or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound or metabolite thereof, or a pharmaceutical composition of the present invention, for use in the prevention and/or treatment of KRAS G12D mediated related diseases.

Another object of the present invention is to provide a use of a compound of the present invention or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound or metabolite, or a pharmaceutical composition of the present invention, for the preparation of a medicament for the prevention and/or treatment of KRAS G12D mediated related diseases.

It is another object of the present invention to provide a method for preventing and or treating KRAS G12D mediated related diseases comprising administering to a subject in need thereof a prophylactically or therapeutically effective amount of a compound of the present invention, or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound or metabolite thereof, or a pharmaceutical composition of the present invention.

According to some embodiments of the invention, the KRAS G12D mediated related disease is a tumor, preferably, the KRAS G12D mediated related disease is cancer.

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

The dosing regimen may be adjusted to provide the best 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 noted that dosage values may vary with the type and severity of the condition being alleviated, and may include single or multiple doses. It is further understood that for any particular individual, the specific dosage regimen will be adjusted over time according to the individual need and the professional judgment of the person administering the composition or supervising the administration of the composition.

The amount of a compound of the invention administered will depend on the subject being treated, the severity of the disorder or condition, the rate of administration, the disposition of the compound, and the judgment of the prescribing physician. In some cases, dosage levels not higher than the lower limit of the aforesaid range may be sufficient, while in other cases still larger doses may be employed without causing any harmful side effects, provided that the larger dose is first divided into several smaller doses to be administered throughout the day.

As used herein, unless otherwise specified, the term "treating" means reversing, alleviating, ameliorating the progression of a disorder or condition to which such term applies or one or more symptoms of such disorder or condition.

The term "prevention" refers to inhibiting and delaying the onset of a disease and includes not only prevention prior to the development of the disease, but also prevention of the recurrence of the disease after treatment.

As used herein, "individual" includes a human or non-human animal. Exemplary human individuals include human individuals (referred to as patients) having a disease (e.g., a disease described herein) or normal individuals. "non-human animals" in the context of 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.).

Detailed Description

In order to make the objects and technical solutions of the present invention clearer, embodiments of the present invention will be described in detail below with reference to examples. It will be understood by those skilled in the art that the following examples are illustrative of the present invention only and should not be taken as limiting the scope of the invention. The examples, in which the specific conditions are not specified, were conducted under the conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products commercially available.

The structure of the compound is determined by nuclear magnetic resonance ¹ H NMR) or Mass Spectrometry (MS). ¹ The H NMR analyzer is a JEOL Eclipse 400 NMR spectrometer, and the measuring solvent is deuterated methanol (CD) ₃ OD), deuterated chloroform (CDCl) ₃ ) Or hexadeutero dimethyl sulfoxide (DMSO-d) ₆ ) Internal standard is Tetramethylsilane (TMS) and chemical shifts (δ) are given in parts per million (ppm).

The measurement instrument for MS was an Agilent (ESI) mass spectrometer, manufacturer: agilent, model: agilent 6120B.

The preparation method of the high performance liquid chromatograph comprises the following steps:

the instrument model is as follows: agilent 1260, column: waters SunAire Prep C18 OBD (19 mm. Times.150 mm. Times.5.0 μm); temperature of the chromatographic column: 25 ℃; flow rate: 20.0mL/min; detection wavelength: 214nm; elution gradient: (0min; a mobile phase A: acetonitrile; mobile phase B:0.05% aqueous formic acid.

Thin layer chromatography silica gel plate (TLC) an aluminum plate (20X 20 cm) from Merck was used, and the specification for separation and purification by thin layer chromatography was GF 254 (1 mm) from Nicotiana.

The reaction was monitored by Thin Layer Chromatography (TLC) or LC-MS; the developer system used included: dichloromethane and methanol system, n-hexane and ethyl acetate system, and petroleum ether and ethyl acetate system, and the volume ratio of the solvent is regulated according to different polarities of the compounds or by adding triethylamine and the like.

The microwave reaction was carried out using a Biotage Initiator + (400W, RT-300 ℃ C.) microwave reactor.

The column chromatography generally uses 200-300 mesh silica gel as a carrier. The eluent system comprises: the volume ratio of the solvent is adjusted according to different polarities of the compounds in a dichloromethane and methanol system and a petroleum ether and ethyl acetate system, and a small amount of triethylamine can be added for adjustment.

In the examples, the reaction temperature is room temperature (20 ℃ to 35 ℃);

the reagents used in the present invention were purchased from Acros Organics, aldrich Chemical Company, texas Chemical, and the like.

In the conventional synthesis methods, examples, and intermediate synthesis examples, the meanings of the abbreviations are as follows.

Examples of preparation of intermediates

Intermediate preparation example 1: preparation of 3-benzyloxy-1-bromonaphthalene

4-Bromomaphthalen-2-ol (5.0g, 22.19mmol) and potassium carbonate (9.20g, 66.57mmo) were sequentially added to acetonitrile (100 mL), and benzyl bromide (4.22g, 24.41mmol) was added dropwise with stirring at room temperature. After dropping, the system was stirred at 80 ℃ for 1 hour. The system was cooled to room temperature and the 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), and the organic phase was dried over anhydrous sodium sulfate and concentrated to dryness to give 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)。

Intermediate preparation example 2: preparation of tert-butyl 3- (2- ((2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidin-4-yl) -8-azabicyclo [3.2.1] octane-8-carboxylate

The first step is as follows: preparation of tert-butyl 3- (7-benzyl-2-chloro-5, 6,7, 8-tetrahydropyrido [3,4-d ] pyrimidin-4-yl) -8-azabicyclo [3.2.1] oct-2-ene-8-carboxylate

7-benzyl-2, 4-dichloro-5, 6,7, 8-tetrahydropyrido [3,4-D ] pyrimidine (5.0g, 17.0mmol), 8-tert-butoxycarbonyl-8-azabicyclo [3.2.1] oct-2-ene-3-boronic acid pinacol ester (8.5g, 25.5mmol), potassium acetate (5.0g, 51mmol) and [1,1' -bis (diphenylphosphino) ferrocene ] dichloropalladium dichloromethane complex (1.24g, 1.7mmol) were added to 1, 4-dioxane (50 mL) and water (10 mL), replaced with nitrogen gas three times, and the reaction system was stirred at 90 ℃ for 12 hours. After the system was cooled to room temperature, the solvent was removed by rotary evaporation, and purified by thin layer chromatography (eluent: petroleum ether/ethyl acetate =2/1, volume ratio) to obtain the title compound (4.96 g, yield: 62.5%).

MS m/z(ESI):467.2[M+H] ⁺ 。

The second step is that: preparation of 3- (7-benzyl-2- ((2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidin-4-yl) -8-azabicyclo [3.2.1] oct-2-ene-8-carboxylic acid tert-butyl ester

Tert-butyl 3- (7-benzyl-2-chloro-5,6,7,8-tetrahydropyrido [3,4-d ] pyrimidin-4-yl) -8-azabicyclo [3.2.1] oct-2-ene-8-carboxylate (4.96g, 10.6 mmol), ((2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methanol (3.38g, 21.2mmol), cesium carbonate (10.36g, 31.8mmol) and methanesulfonic acid (2-dicyclohexylphosphino-2 ',6' -diisopropoxy-1, 1 '-biphenyl) (2-amino-1, 1' -biphenyl-2-yl) palladium (II) (0.89g, 1.06mmol) were sequentially added to toluene (80 mL), replaced three times with nitrogen, and the reaction was stirred at 110 ℃ for 4 hours. The system was cooled to room temperature, the solvent was removed by rotary evaporation, and purified by silica gel column chromatography (eluent: dichloromethane/methanol =10/1, volume ratio) to obtain the title compound (3.2 g, yield: 51.0%).

MS m/z(ESI):590.3[M+H] ⁺ 。

The third step: preparation of tert-butyl 3- (2- ((2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidin-4-yl) -8-azabicyclo [3.2.1] octane-8-carboxylate

Tert-butyl 3- (7-benzyl-2- ((2r, 7as) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidin-4-yl) -8-azabicyclo [3.2.1] oct-2-ene-8-carboxylate (3.2g, 5.42mmol) was dissolved in a methanolic ammonia solution (50 mL), 10% palladium on carbon (0.8 g) was added, the mixture was replaced with hydrogen three times, and the reaction system was stirred at 45 ℃ for 24 hours under a hydrogen atmosphere. The system was cooled to room temperature, filtered, and concentrated to give the title compound (2.58 g, yield: 94.8%).

MS m/z(ESI):502.3[M+H] ⁺ 。

Intermediate preparation example 3: preparation of 1, 8-dibromo-3- (methoxymethoxy) naphthalene

The first step is as follows: preparation of 2- (4, 5-dibromo-2-naphthyl) -4, 5-tetramethyl-1, 3, 2-dioxaborolan

1, 8-dibromonaphthalene (1 g, 3.46mmol), bis (1, 5-cyclooctadiene) bis-m-methoxydiidium (I) (114.49mg, 173.10. Mu. Mol) and 4,4 '-di-tert-butyl-2, 2' -bipyridine (56.31mg, 207.72. Mu. Mol) were dissolved in tetrahydrofuran (15.0 mL), replaced with nitrogen three times, 4, 5-tetramethyl-1, 3, 2-dioxaborolane (4.48g, 34.62mmol) was added with stirring, and the system was stirred at 70 ℃ for 10 hours. After the reaction solution was cooled to room temperature, the reaction was quenched by dropping water (30 mL) into the reaction solution, extracted twice with ethyl acetate (30 mL), the organic phases were combined, dried over anhydrous sodium sulfate, the solvent was removed by rotary evaporation, and the residue was purified by silica gel column chromatography (eluent: petroleum ether) to give the title compound (648 mg, yield: 22.7%).

MS m/z(ESI):413.0[M+H] ⁺ 。

The second step is that: preparation of 4, 5-dibromonaphthalen-2-ol

2- (4, 5-dibromo-2-naphthyl) -4, 5-tetramethyl-1, 3, 2-dioxaborolane (654 mg, 1.11mmol) was dissolved in tetrahydrofuran (5.08 mL), water (17 mL) was added, acetic acid (5.17g, 71.35mmol) and 30% hydrogen peroxide (2.41g, 21.22mmol) were added with stirring at 10 ℃, and the reaction system was reacted at 10 ℃ for 15 hours. To the reaction solution was added a sodium hydrogen sulfite solution (20 mL), extracted twice with ethyl acetate (30 mL), the organic phases were combined, dried over anhydrous sodium sulfate, the solvent was removed by rotary evaporation, and the residue was purified by silica gel column chromatography (eluent: petroleum ether/ethyl acetate =4/1, volume ratio) to obtain the title compound (268 mg, yield: 75.87%).

MS m/z(ESI):303.1[M+H] ⁺ 。

The third step: preparation of 1, 8-dibromo-3- (methoxymethoxy) naphthalene

4, 5-Dibromonaphthalen-2-ol (162mg, 509.67. Mu. Mol) was dissolved in dichloromethane (3.5 mL), and bromo (methoxy) methane (193.69mg, 1.53mmol) and N, N-diisopropylethylamine (333.14mg, 2.55mmol) were added with stirring at 0 ℃ and reacted at 0 ℃ for 1 hour. Water (10 mL) was added to the reaction solution, extraction was performed with dichloromethane (20 mL), the organic phase was washed twice with water (20 mL), the organic phases were combined, dried over anhydrous sodium sulfate, the solvent was removed by rotary evaporation, and the residue was purified by silica gel column chromatography (eluent: petroleum ether/ethyl acetate =9/1, volume ratio) to obtain the title compound (160 mg, yield: 81.65%).

MS m/z(ESI):347.1[M+H] ⁺ 。

¹ H-NMR(400MHz,CDCl ₃ ):δ7.72(dd,J＝8.0Hz,J＝4.0Hz,1H),7.64(dd,J＝8.0Hz,J＝4.0Hz,2H),7.32(d,J＝4.0Hz,1H),7.16-7.11(m,1H),5.20(s,2H),3.44(s,3H)。

Intermediate preparation example 4: preparation of tert-butyl 3- (7-chloro-8-fluoro-2- ((2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) pyridine [4,3-d ] pyrimidin-4-yl) -8-azabicyclo [3.2.1] octane-8-carboxylate

The first step is as follows: preparation of 2, 7-dichloro-8-fluoro-4-methoxypyridine [4,3-d ] pyrimidine

2,4, 7-trichloro-8-fluoropyridine [4,3-d ] pyrimidine (5.0g, 19.8mmol) was dissolved in methanol (50.0 mL), nitrogen gas was substituted three times, the system was cooled to 0 ℃ and sodium methoxide (1.07g, 19.8mmol) was added in portions, the reaction mixture was warmed to room temperature after the addition was stirred for 1 hour, the reaction mixture was quenched by dropping water (30 mL), extracted twice with ethyl acetate (100 mL), the organic phases were combined, washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, the solvent was removed by rotary evaporation, and the residue was purified by silica gel column chromatography (eluent: petroleum ether/ethyl acetate =10/1, volume ratio) to give the title compound (2.1 g, yield: 42.7%).

MS m/z(ESI):248.0[M+H] ⁺ 。

The second step is that: preparation of 7-chloro-8-fluoro-2- ((2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -4-methoxypyridine [4,3-d ] pyrimidine

2, 7-dichloro-8-fluoro-4-methoxypyridine [4,3-d ] pyrimidine (2.1g, 8.5 mmol) was dissolved in acetonitrile (30 mL), N-diisopropylethylamine (3.3g, 25.5 mmol) and ((2R, 7 aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methanol (4.06g, 25.5 mmol) were added, and the reaction system was reacted at 80 ℃ for 5 hours. The reaction mixture was cooled to room temperature, and a solid precipitated, which was filtered and the cake was dried to obtain the title compound (2.0 g, yield: 63.7%).

MS m/z(ESI):371.1[M+H] ⁺ 。

The third step: preparation of 7-chloro-8-fluoro-2- ((2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) pyridine [4,3-d ] pyrimidin-4-ol

7-chloro-8-fluoro-2- ((2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -4-methoxypyridine [4,3-d ] pyrimidine (2.0 g, 5.39mmol) was dissolved in acetonitrile (30 mL), and trimethyliodosilane (5.39g, 26.95mmol) was added dropwise at room temperature, and the reaction was carried out at room temperature for 12 hours. The reaction was quenched by adding water (50 mL) to the reaction solution, extracted four times with dichloromethane (20 mL), the organic phase was washed twice with aqueous sodium sulfite solution (20 mL), dried over anhydrous sodium sulfate, the solvent was removed by rotary evaporation, and the residue was purified by silica gel column chromatography (eluent: dichloromethane/methanol =10/1, volume ratio) to obtain the title compound (1.1 g, yield: 57.3%).

MS m/z(ESI):357.1[M+H] ⁺ 。

The fourth step: preparation of 4, 7-dichloro-8-fluoro-2- ((2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) pyridine [4,3-d ] pyrimidine

7-chloro-8-fluoro-2- ((2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) pyridin [4,3-d ] pyrimidin-4-ol (1.1g, 3.08mmol) was dissolved in thionyl chloride (30 mL), 1 drop of DMF was added dropwise at room temperature, and the reaction was carried out at 90 ℃ for 6 hours. The reaction was cooled to room temperature, concentrated, and the solid residue was diluted with ethyl acetate (20 mL), washed with a sodium bicarbonate solution (50 mL) under an ice bath, separated, the organic phase dried over anhydrous sodium sulfate, and the solvent removed by rotary evaporation to give the title compound (0.8 g, yield: 68.9%).

MS m/z(ESI):375.1[M+H] ⁺ 。

The fifth step: preparation of 3- (7-chloro-8-fluoro-2- ((2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) pyridine [4,3-d ] pyrimidin-4-yl) -8-azabicyclo [3.2.1] oct-2-ene-8-carboxylic acid tert-butyl ester

4, 7-dichloro-8-fluoro-2- ((2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) pyridine [4,3-d ] pyrimidine (0.8g, 2.13mmol), 8-tert-butoxycarbonyl-8-azabicyclo [3.2.1] oct-2-ene-3-boronic acid pinacol ester (0.71g, 2.13mmol), potassium acetate (0.42g, 4.26mmol) and [1,1' -bis (diphenylphosphino) ferrocene ] palladium dichloride dichloromethane complex (0.15g, 0.21mmol) were added to 1, 4-dioxane (10 mL) and water (1 mL), replaced with nitrogen three times, and the reaction system was stirred at 90 ℃ for 1 hour. After the system was cooled to room temperature, the solvent was removed by rotary evaporation, and purified by thin layer chromatography (eluent: ethyl acetate/methanol =30/1, volume ratio) to obtain the title compound (0.16 g, yield: 13.7%).

MS m/z(ESI):548.2[M+H] ⁺ 。

And a sixth step: preparation of tert-butyl 3- (7-chloro-8-fluoro-2- ((2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) pyridine [4,3-d ] pyrimidin-4-yl) -8-azabicyclo [3.2.1] octane-8-carboxylate

Tert-butyl 3- (7-chloro-8-fluoro-2- ((2r, 7as) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) pyridine [4,3-d ] pyrimidin-4-yl) -8-azabicyclo [3.2.1] oct-2-ene-8-carboxylate (0.16g, 0.29mmol) was dissolved in methanol (3 mL), cobalt chloride hexahydrate (6.9mg, 0.029mmol) was added, the reaction system was cooled to 0 ℃, sodium borohydride (22.04mg, 0.58mmol) was added, stirring was performed at room temperature for 2 hours, the reaction was quenched by dropping water (10 mL) into the reaction solution, extraction was performed twice with ethyl acetate (20 mL), the organic phases were combined, the organic phase was washed with saturated brine (100 mL), dried with anhydrous sodium sulfate, the solvent was removed by rotary evaporation, and purification was performed by thin layer chromatography (eluent: ethyl acetate/dichloromethane =2/1, volume ratio) to obtain the title compound (80 mg, yield: 49.7%).

MS m/z(ESI):550.2[M+H] ⁺ 。

Intermediate preparation example 5: preparation of 1-bromo-8-chloro-3- (methoxymethoxy) naphthalene

The first step is as follows: preparation of 2, 4-dibromo-5-chloronaphthalene-1-amine

5-chloronaphthalene-1-amine (1g, 5.35mmol) was dissolved in chloroform (10 mL), and bromine (1.71g, 10.7 mmol) was added dropwise with stirring, and after a reaction at 50 ℃ for 15 hours, bromine (1.71g, 10.7 mmol) was added dropwise and reacted for 5 hours. The reaction solution was concentrated, and the residue was purified by column chromatography on silica gel (eluent: petroleum ether) to give the title compound (1.28 g, yield: 71.5%).

MS m/z(ESI):336.0[M+H] ⁺ 。

The second step: preparation of 5-bromo-6-chloronaphthalene [1,2-d ] [1,2,3] oxadiazole

2, 4-dibromo-5-chloronaphthalene-1-amine (1.28g, 3.82mmol) was dissolved in acetic acid (10 mL) and propionic acid (2 mL), cooled to 0 deg.C, and sodium nitrite (315.9 mg, 4.58mmol) was slowly added and reacted for 1 hour. Water (30 mL) was added to the reaction solution to give a turbid solution, which was filtered, the filter cake was washed with water (20 mL), and the filter cake was dried to give the title compound (564 mg, yield: 58.1%).

MS m/z(ESI):285.0[M+H] ⁺ 。

The third step: preparation of 4-bromo-5-chloronaphthalen-2-ol

5-bromo-6-chloronaphthalene [1,2-d ] [1,2,3] oxadiazole (564mg, 1.99mmol) was dissolved in tetrahydrofuran (3 mL) and ethanol (6 mL), cooled to 0 deg.C, and added with sodium borohydride (150.5mg, 3.98mmol) to react for 1.5 hours. The reaction solution was concentrated, and the residue was purified by silica gel column chromatography (eluent: petroleum ether/ethyl acetate =6/1, volume ratio) to obtain the title compound (170 mg, yield: 31.2%).

MS m/z(ESI):259.0[M+H] ⁺ 。

The fourth step: preparation of 1-bromo-8-chloro-3- (methoxymethoxy) naphthalene

4-bromo-5-chloronaphthalen-2-ol (150mg, 582.5. Mu. Mol) was dissolved in dichloromethane (10 mL), cooled to 0 ℃ and bromomethyl methyl ether (218.4mg, 1.75mmol), N, N-diisopropylethylamine (376.4mg, 2.91mmol) was added slowly and reacted for 1 hour. To the reaction solution, water (10 mL) was added, extraction was performed with ethyl acetate (20 mL), the organic phases were backwashed with saturated brine (20 mL), the combined organic phases were dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (eluent: petroleum ether/ethyl acetate =10/1, volume ratio) to obtain the title compound (79 mg, yield: 44.1%).

MS m/z(ESI):303.0[M+H] ⁺ 。

Intermediate preparation example 6: preparation of 8-chloro-7-fluoronaphthalen-1-yl trifluoromethanesulfonate

The first step is as follows: preparation of 5-chloro-6-fluoro-1, 4-dihydro-1, 4-epoxynaphthalene

1-bromo-3-chloro-2, 4-difluorobenzene (1.0 g, 4.40mmol) and furan (897.90mg, 13.19mmol) were added to diethyl ether (10.0 mL), the mixture was cooled to-70 ℃ after three nitrogen replacements, n-butyllithium (366.42mg, 5.72mmol) was added while maintaining the low temperature, and then the reaction was continued at that temperature for 1 hour, whereupon the reaction was allowed to proceed to 20 ℃ and stirring was continued for 12 hours. The reaction mixture was quenched by dropping water (30 mL) thereto, extracted twice with ether (30 mL), the organic phases were combined, dried over anhydrous sodium sulfate, and the solvent was removed by rotary evaporation to give the title compound (600 mg, yield: 69.4%) as a yellow oil.

MS m/z(ESI):197.0[M+H] ⁺ 。

The second step is that: preparation of 8-chloro-7-fluoronaphthalen-1-ol

5-chloro-6-fluoro-1, 4-dihydro-1, 4-epoxynaphthalene (600mg, 3.05mmol) was dissolved in ethanol (10.0 mL), and concentrated hydrochloric acid (5 mL) was added thereto, followed by reaction at 80 ℃ for 2 hours with stirring. The reaction solution was concentrated, and the residue was purified by column chromatography on silica gel (eluent: petroleum ether/ethyl acetate =10/1, volume ratio) to give the title compound (300 mg, yield: 50.0%) as a pale yellow oil.

MS m/z(ESI):197.0[M+H] ⁺ 。

The third step: preparation of 8-chloro-7-fluoronaphthalen-1-yl trifluoromethanesulfonate

8-chloro-7-fluoronaphthalen-1-ol (300mg, 1.53mmol) and N, N-diisopropylethylamine (1.18g, 9.16mmol) were dissolved in dichloromethane (6 mL), and trifluoromethanesulfonic anhydride (648.92mg, 2.30mmol) was added with stirring at 0 ℃ under nitrogen protection and reacted at 0 ℃ for 1 hour. Water (10 mL) was added to the reaction solution, extracted twice with dichloromethane (20 mL), the organic phases were combined, the organic phase was washed twice with water (20 mL), dried over anhydrous sodium sulfate, the solvent was removed by rotary evaporation, and the residue was purified by silica gel column chromatography (eluent: petroleum ether/ethyl acetate =15/1, volume ratio) to obtain the title compound (400 mg, yield: 79.76%).

¹ H NMR(400MHz,CDCl ₃ ):δ7.89(dd,J＝8.0Hz,J＝1.2Hz,1H),7.83(dd,J＝9.2Hz,J＝5.2Hz,1H),7.58(d,J＝7.6Hz,1H),7.50(t,J＝8.0Hz,1H),7.43(t,J＝8.0Hz,1H)。

Examples

Example 1:4- (4- ((1R, 3s, 5S) -8-azabicyclo [3.2.1] octan-3-yl) -2- ((S) -1-methylpyrrolidin-2-yl) methoxy) -5, 8-dihydropyrido [3,4-d ] pyrimidin-7 (8H) -yl) naphthalen-2-ol (compound 1A or 1B) and

preparation of 4- (4- ((1R, 3r, 5S) -8-azabicyclo [3.2.1] octan-3-yl) -2- ((S) -1-methylpyrrolidin-2-yl) methoxy) -5, 8-dihydropyrido [3,4-d ] pyrimidin-7 (8H) -yl) naphthalen-2-ol (Compound 1A or 1B)

The first step is as follows: preparation of 7-benzyl-2-chloro-4-methoxy-5, 6,7, 8-tetrahydropyrido [3,4-d ] pyrimidine

7-benzyl-2, 4-dichloro-5, 6,7, 8-tetrahydropyrido [3,4-d ] pyrimidine (2.0 g, 6.73mmol) was dissolved in anhydrous methanol (30 mL), cooled in an ice-salt bath, sodium methoxide (385.65mg, 7.07mmol) was added in portions at 0 ℃ and, after completion of addition, the reaction was stirred at 0 ℃ for 3 hours. The reaction system was quenched by dropwise addition of ice water (50 mL), extracted three times with ethyl acetate (20 mL), the organic phases were combined, back-washed twice with saturated brine (20 mL), dried over anhydrous sodium sulfate, and the solvent was rotary evaporated under reduced pressure to give the title compound (1.91 g, yield: 93%).

MS m/z(ESI):290.1[M+H] ⁺ 。

The second step is that: preparation of (S) -7-benzyl-4-methoxy-2- ((1-methylpyrrolidin-2-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidine

7-benzyl-2-chloro-4-methoxy-5, 6,7, 8-tetrahydropyrido [3,4-d ] pyrimidine (1.9g, 6.23mmol), (S) - (1-methylpyrrolidin-2-yl) methanol (942.10mg, 8.10mmol), cesium carbonate (5.13g, 15.57mmol), 4, 5-bis-diphenylphosphine-9, 9-dimethylxanthene (364.08mg, 622.93. Mu. Mol), and [1,1' -bis (diphenylphosphine) ferrocene ] palladium dichloride dichloromethane complex (230.20mg, 311.47. Mu. Mol) were added to 1, 4-dioxane (30 mL), replaced three times with nitrogen, and the reaction system was stirred at 100 ℃ for 1 hour. After the system was cooled to room temperature, the solvent was rotary-evaporated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent: dichloromethane/methanol =10/1, volume ratio) to obtain the title compound (2.1 g, yield: 86.9%).

MS m/z(ESI):369.3[M+H] ⁺ 。

The third step: preparation of (S) -4-methoxy-2- ((1-methylpyrrolidin-2-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidine

10% Palladium on carbon (0.5 g) and (S) -7-benzyl-4-methoxy-2- ((1-methylpyrrolidin-2-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidine (2.0 g, 5.16mmol) were added to anhydrous methanol (30 mL), replaced three times with hydrogen gas, and the reaction system was stirred at room temperature for 5 hours. The filter cake was washed with methanol (50 mL) by suction filtration, and the solvent was removed from the filtrate by rotary evaporation under reduced pressure to give the title compound (1.15 g, yield: 76.12%).

MS m/z(ESI):279.3[M+H] ⁺ 。

The fourth step: preparation of (S) -7- (3- (benzyloxy) naphthalen-1-yl) -4-methoxy-2- ((1-methylpyrrolidin-2-yl) methoxy) -5,6,7, 8-tetrahydropyridine [3,4-d ] pyrimidine

(S) -4-methoxy-2- ((1-methylpyrrolidin-2-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidine (1.1g, 3.75mmol), 3- (benzyloxy) -1-bromonaphthalene (1.78g, 5.63mmol), cesium carbonate (3.09g, 9.39mmol), 2-dicyclohexylphosphorus-2 ',6' -diisopropoxy-1, 1' -biphenyl (177mg, 0.537mmol) and tris (dibenzylideneacetone) dipalladium (173mg, 0.187mmol) were added to 1, 4-dioxane (15 mL), replaced three times with nitrogen, and the reaction system was stirred at 100 ℃ for 8 hours. After the system was cooled to room temperature, the solvent was rotary-evaporated under reduced pressure, and the residue was purified by column chromatography on silica gel (eluent: dichloromethane/methanol =10/1, volume ratio) to obtain the title compound (910 mg, yield: 45.1%).

MS m/z(ESI):511.3[M+H] ⁺ 。

The fifth step: preparation of (S) -7- (3- (benzyloxy) naphthalen-1-yl) -2- ((1-methylpyrrolidin-2-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidin-4-ol

(S) -7- (3- (benzyloxy) naphthalen-1-yl) -4-methoxy-2- ((1-methylpyrrolidin-2-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidine (900mg, 1.67mmol) and sodium ethanethiol (569.10mg, 6.70mmol) were added to DMF (5 mL), replaced three times with nitrogen, and the reaction system was stirred at 100 ℃ for 2 hours. After the system was cooled to room temperature, and purified by high performance liquid chromatography, the title compound (370 mg, yield: 42.27%) was obtained.

MS m/z(ESI):497.2[M+H] ⁺ 。

¹ H-NMR(400MHz,CD ₃ OD):δ8.05(t,J＝8.0Hz,1H),7.80-7.74(m,1H),7.51-7.44(m,3H),7.41-7.31(m,4H),7.18-7.11(m,1H),7.02-6.91(m,1H),5.20(s,2H),4.61-4.56(m,1H),4.21(m,1H),3.95-3.70(m,3H),3.50-3.48(m,1H),3.27-3.21(m,1H),3.07(s,3H),3.02-2.95(m,1H),2.78-2.60(m,2H),2.44-2.36(m,1H),2.25-1.95(m,4H)。

And a sixth step: preparation of (S) -7- (3- (benzyloxy) naphthalen-1-yl) -2- ((1-methylpyrrolidin-2-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidin-4-yl trifluoromethanesulfonate

(S) -7- (3- (benzyloxy) naphthalen-1-yl) -2- ((1-methylpyrrolidin-2-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidin-4-ol (150mg, 267.83. Mu. Mol) was dissolved in dichloromethane (5 mL), cooled to-40 ℃ and trifluoromethanesulfonic anhydride (114.49mg, 401.75. Mu. Mol) was added dropwise at-40 ℃ and the reaction was stirred at-40 ℃ for 1 hour. Quenched by dropwise addition of 1 drop of ammonium chloride solution at-40 ℃, the solvent was removed by rotary evaporation under reduced pressure, and purified by thin layer chromatography (eluent: dichloromethane/methanol =10/1, volume ratio) to obtain the title compound (103 mg, yield: 58.1%).

MS m/z(ESI):629.2[M+H] ⁺ 。

The seventh step: preparation of 3- (7- (3- (benzyloxy) naphthalen-1-yl) -2- (((S) -1-methylpyrrolidin-2-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidin-4-yl) -8-azabicyclo [3.2.1] oct-2-ene-8-carboxylic acid tert-butyl ester

(S) -7- (3- (benzyloxy) naphthalen-1-yl) -2- ((1-methylpyrrolidin-2-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidin-4-yl trifluoromethanesulfonate (113mg, 170.76. Mu. Mol), tert-butyl 3- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -8-azabicyclo [3.2.1] oct-2-ene-8-carboxylate (115.65mg, 341.52. Mu. Mol), sodium carbonate (45.70mg, 426.90. Mu. Mol) and [1,1' -bis (diphenylphosphino) ferrocene ] dichloropalladium dichloromethane complex (10.10mg, 13.66. Mu. Mol) were added to 1, 4-dioxane (5 mL), replaced with nitrogen three times, and the reaction system was stirred at 100 ℃ for 12 hours. After the system was cooled to room temperature, the solvent was rotovaped off under reduced pressure, and purified by thin layer chromatography (eluent: dichloromethane/methanol =10/1, volume ratio) to obtain the title compound (108 mg, yield: 87.3%).

MS m/z(ESI):688.3[M+H] ⁺ 。

Eighth step: preparation of tert-butyl 3- (7- (3- (benzyloxy) naphthalen-1-yl) -2- (((S) -1-methylpyrrolidin-2-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidin-4-yl) -8-azabicyclo [3.2.1] octane-8-carboxylate

10% Palladium on carbon (15mg, 89.77. Mu. Mol) and tert-butyl 3- (7- (3- (benzyloxy) naphthalen-1-yl) -2- (((S) -1-methylpyrrolidin-2-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidin-4-yl) -8-azabicyclo [3.2.1] oct-2-ene-8-carboxylate (65mg, 89.77. Mu. Mol) were added to anhydrous methanol (4 mL), replaced three times with hydrogen, the reaction was stirred at room temperature for 2 hours, filtered under suction, the cake was washed with methanol (10 mL) under reduced pressure, and the solvent was rotary evaporated to give the title compound (58 mg, yield: 89.0%).

MS m/z(ESI):690.4[M+H] ⁺ 。

The ninth step: preparation of 4- (4- (8-azabicyclo [3.2.1] octan-3-yl) -2- ((S) -1-methylpyrrolidin-2-yl) methoxy) -5, 8-dihydropyrido [3,4-d ] pyrimidin-7 (8H) -yl) naphthalen-2-ol (Compound 1)

Tert-butyl 3- (7- (3- (benzyloxy) naphthalen-1-yl) -2- (((S) -1-methylpyrrolidin-2-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidin-4-yl) -8-azabicyclo [3.2.1] octane-8-carboxylate (58mg, 79.87. Mu. Mol) was dissolved in dichloromethane (3 mL), cooled in an ice-water bath, and the hydrobromic acid/acetic acid solution (107.71mg, 399.34. Mu. Mol) was added dropwise at 0 ℃ and the reaction was stirred at 0 ℃ for 0.5 h. The reaction system was separated to remove the supernatant, the solid was adjusted to pH =7 with sodium bicarbonate solution, extracted three times with ethyl acetate (10 mL), the organic phases were combined, back-washed twice with saturated brine (10 mL), dried over anhydrous sodium sulfate, and the solvent was rotary evaporated under reduced pressure to give the title compound (30 mg, yield: 75.2%).

MS m/z(ESI):500.3[M+H] ⁺ 。

The tenth step: 4- (4- ((1R, 3s, 5S) -8-azabicyclo [ 3.2.1)]Octane-3-yl) -2- ((S) -1-methylpyrrolidin-2-yl) methoxy) -5, 8-dihydropyrido [3,4-d]Pyrimidin-7 (8H) -yl) naphthalen-2-ol(Compound 1A or 1B)And

4- (4- ((1R, 3r, 5S) -8-azabicyclo [ 3.2.1)]Octane-3-yl) -2- ((S) -1-methylpyrrolidin-2-yl) methoxy) -5, 8-dihydropyrido [3,4-d]Pyrimidin-7 (8H) -yl) naphthalen-2-ol(Compound 1A or 1B)Preparation of (2)

The crude product of the ninth step was purified by high performance liquid chromatography to give the title compound 1A (2.10 mg, yield: 4.58%) and compound 1B (2.56 mg, yield: 5.58%).

Retention time for compound 1A was 7.0 minutes;

MS m/z(ESI):500.3[M+H] ⁺ ；

¹ H-NMR(400MHz,CD ₃ OD):δ8.54(br,1H),8.03(d,J＝8.0Hz,1H),7.64(d,J＝8.0Hz,1H),7.37(dt,J＝8.0Hz,J＝4.0Hz,1H),7.25(dt,J＝8.0Hz,J＝4.0Hz,1H),6.87(d,J＝4.0Hz,1H),6.79(d,J＝4.0Hz,1H),4.42(d,J＝8.0Hz,1H),4.22(s,2H),4.04(s,2H),3.53-3.44(m,3H),3.14-3.09(m,2H),2.99-2.97(m,2H),2.83-2.81(m,1H),2.54-2.43(m,5H),2.43-2.29(m,5H),2.15-2.10(m,1H),2.06-2.00(m,2H),1.86-1.82(m,2H),1.77-1.69(m,1H)。

retention time of compound 1B was 7.8 minutes;

MS m/z(ESI):500.3[M+H] ⁺ ；

¹ H-NMR(400MHz,CD ₃ OD):δ8.52(br,1H),8.03(d,J＝8.0Hz,1H),7.64(d,J＝8.0Hz,1H),7.37(dt,J＝8.0Hz,J＝4.0Hz,1H),7.25(dt,J＝8.0Hz,J＝4.0Hz,1H),6.87(d,J＝4.0Hz,1H),6.79(d,J＝4.0Hz,1H),4.51-4.42(m,2H),4.22(s,2H),4.16(s,2H),3.58-3.50(m,3H)3.15-3.03(m,4H),2.65-2.60(m,4H),2.34-2.15(m,7H),1.97-1.89(m,4H),1.83-1.74(m,1H)。

example 2:4- ((1R, 3s, 5S) -8-azabicyclo [3.2.1] octan-3-yl) -7- (8-chloronaphthalen-1-yl) -2- ((2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidine (compound 29A or 29B) and

preparation of 4- ((1R, 3r, 5S) -8-azabicyclo [3.2.1] octan-3-yl) -7- (8-chloronaphthalen-1-yl) -2- ((2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidine (Compound 29A or 29B)

The first step is as follows: preparation of 3- (7- (8-chloronaphthalen-1-yl) -2- ((2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidin-4-yl) -8-azabicyclo [3.2.1] octane-8-carboxylic acid tert-butyl ester

Tert-butyl 3- (2- ((2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidin-4-yl) -8-azabicyclo [3.2.1] octane-8-carboxylate (100mg, 0.19mmol), 1-bromo-8-chloronaphthalene (96.29mg, 0.38mmol), cesium carbonate (194.85mg, 0.57mmol), 2-dicyclohexylphosphonium-2 ',6' -diisopropoxy-1, 1' -biphenyl (22.10mg, 38.77. Mu. Mol) and tris (dibenzylideneacetone) dipalladium (26.54mg, 28.41. Mu. Mol) were added to 1, 4-dioxane (3 mL), replaced three times with nitrogen, and the reaction system was stirred at 95 ℃ for 8 hours. After the system was cooled to room temperature, the solvent was rotovapped off under reduced pressure, and the residue was purified by silica gel column chromatography (eluent: dichloromethane/methanol =10/1, volume ratio) to obtain the title compound (97 mg, yield: 69.61%).

MS m/z(ESI):662.4[M+H] ⁺ 。

The second step: preparation of 4- (-8-azabicyclo [3.2.1] oct-3-yl) -7- (8-chloronaphthalen-1-yl) -2- ((2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidine (Compound 29)

Tert-butyl 3- (7- (8-chloronaphthalen-1-yl) -2- ((2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidin-4-yl) -8-azabicyclo [3.2.1] octane-8-carboxylate (97.0mg, 131.83. Mu. Mol) was dissolved in dichloromethane (3 mL), trifluoroacetic acid (1 mL) was added dropwise at a temperature of 25 ℃, and the reaction was stirred at 25 ℃ for 1.5 hours. The reaction was concentrated, the residue was diluted with ethyl acetate (3 mL), washed three times with sodium bicarbonate solution (10 mL), and the organic phases were combined, back-washed twice with saturated brine (10 mL), dried over anhydrous sodium sulfate, and the solvent was rotary evaporated under reduced pressure to give the title compound (40.0 mg, crude).

MS m/z(ESI):563.3[M+H] ⁺ 。

The third step: 4- ((1R, 3s, 5S) -8-azabicyclo [3.2.1] octan-3-yl) -7- (8-chloronaphthalen-1-yl) -2- ((2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidine (compound 29A or 29B) and

The crude product of the second step was purified by high performance liquid chromatography to give the title compound 29A (18.0 mg, yield: 21.33%) and compound 29B (10.0 mg, yield: 11.85%).

Compound 29A: retention time 5.72 minutes;

MS m/z(ESI):563.3[M+H] ⁺ 。

¹ H-NMR(400MHz,DMSO-d ₆ ):δ7.93(d,J＝8.0Hz,1H),7.75(d,J＝8.0Hz,1H),7.60-7.51(m,2H),7.45(t,J＝7.6Hz,1H),7.36(d,J＝7.6Hz,1H),5.27(d,J＝4.0Hz,2H),4.19-3.95(m,5H),3.89(s,1H),3.56(d,J＝10Hz,1H),3.42(s,1H),3.17-2.98(m,5H),2.82(dd,J＝12Hz,J＝8.4Hz,1H),2.68(d,J＝14.4Hz,1H),2.12-1.71(m,13H)。

compound 29B: retention time was 6.11 minutes;

MS m/z(ESI):563.3[M+H] ⁺ 。

¹ H-NMR(400MHz,DMSO-d ₆ ):δ7.93(dd,J＝8.0Hz,J＝1.0Hz,1H),7.75(d,J＝8.0Hz,1H),7.56(dd,J＝16.4Hz,J＝8.0Hz,2H),7.45(t,J＝7.6Hz,1H),7.37(d,J＝7.6Hz,1H),5.27(d,J＝12.0Hz,2H),4.21-3.88(m,7H),3.57(d,J＝7.2Hz,1H),3.43-3.29(m,2H),3.19-3.11(m,2H),3.01(d,J＝5.0Hz,1H),2.91-2.78(m,2H),2.22-1.90(m,9H),1.86-1.73(m,4H)。

example 3:4- (4- ((1R, 3s, 5S) -8-azabicyclo [3.2.1] octan-3-yl) -2- ((2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -5, 6-dihydropyrido [3,4-d ] pyrimidin-7 (8H) -yl) -5-fluoronaphthalen-2-ol (compound 32A or 32B) and

preparation of 4- (4- ((1R, 3r, 5S) -8-azabicyclo [3.2.1] octan-3-yl) -2- ((2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -5, 6-dihydropyrido [3,4-d ] pyrimidin-7 (8H) -yl) -5-fluoronaphthalen-2-ol (Compound 32A or 32B)

The first step is as follows: preparation of tert-butyl 3- (7- (8-fluoro-3- (methoxymethyloxy) naphthalen-1-yl) -2- ((2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidin-4-yl) -8-azabicyclo [3.2.1] octane-8-carboxylate

Using the synthetic route of example 2, the first-step reaction starting material 1-bromo-8-chloronaphthalene was replaced with 8-fluoro-3- (methoxymethoxy) naphthalen-1-yl trifluoromethanesulfonate to give the title compound (60 mg, yield: 44.41%).

MS m/z(ESI):706.4[M+H] ⁺ 。

The second step: preparation of 4- (4- (-8-azabicyclo [3.2.1] oct-3-yl) -2- ((2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -5, 6-dihydropyrido [3,4-d ] pyrimidin-7 (8H) -yl) -5-fluoronaphthalen-2-ol (compound 32)

Using the synthetic route of example 2, the second reaction starting material, tert-butyl 3- (7- (8-chloronaphthalen-1-yl) -2- ((2r, 7as) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidin-4-yl) -8-azabicyclo [3.2.1] octane-8-carboxylate, was replaced with tert-butyl 3- (7- (8-fluoro-3- (methoxymethoxy) naphthalen-1-yl) -2- ((2r, 7as) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidin-4-yl) -8-azabicyclo [3.2.1] octane-8-carboxylate to give the title compound (42.1 mg, crude).

MS m/z(ESI):562.3[M+H] ⁺ 。

The third step: preparation of 4- (4- ((1R, 3s, 5S) -8-azabicyclo [3.2.1] octan-3-yl) -2- ((2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -5, 6-dihydropyridin [3,4-d ] pyrimidin-7 (8H) -yl) -5-fluoronaphthalen-2-ol (Compound 32A or 32B)

The crude product of the second step was purified by high performance liquid chromatography to give the title compound 32A (5.0 mg, yield: 9.6%) and compound 32B (6.1 mg, yield: 11.5%).

Compound 32A: the retention time was 4.65 minutes;

MS m/z(ESI):562.3[M+H] ⁺ 。

¹ H-NMR(400MHz,DMSO-d ₆ ):δ7.49(d,J＝8.0Hz,1H),7.36-7.31(m,1H),6.99-6.94(m,1H),6.88(s,1H),6.76(d,J＝4.0Hz,1H),5.35(s,1H),5.21(s,1H),4.13-3.98(m,5H),3.62-3.41(m,3H),3.08-2.67(m,9H),2.33-1.94(m,8H),1.87-1.74(m,5H)。

compound 32B: the retention time was 5.31 minutes;

MS m/z(ESI):562.3[M+H] ⁺ 。

¹ H-NMR(400MHz,DMSO-d ₆ ):δ7.49(d,J＝8.0Hz,1H),7.36-7.31(m,1H),6.99-6.94(m,1H),6.88(s,1H),6.76(d,J＝4.0Hz,1H),5.35(s,1H),5.21(s,1H),4.12-3.93(m,7H),3.08-2.82(m,8H),2.13-1.98(m,10H),1.88-1.76(m,5H)。

example 4:4- (4- ((1R, 3s, 5S) -8-azabicyclo [3.2.1] octan-3-yl) -2- ((2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -5, 6-dihydropyrido [3,4-d ] pyrimidin-7 (8H) -yl) -5, 6-difluoronaphthalen-2-ol (compound 33A or 33B) and

preparation of 4- (4- ((1R, 3r, 5S) -8-azabicyclo [3.2.1] octan-3-yl) -2- ((2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -5, 6-dihydropyrido [3,4-d ] pyrimidin-7 (8H) -yl) -5, 6-difluoronaphthalen-2-ol (Compound 33A or 33B)

The first step is as follows: preparation of tert-butyl 3- (7, 8-difluoro-3- (methoxymethyloxy) naphthalen-1-yl) -2- ((2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidin-4-yl) -8-azabicyclo [3.2.1] octane-8-carboxylate

Using the synthetic route of example 2, the reaction starting material 1-bromo-8-chloronaphthalene of the first step was replaced with 7,8-difluoro-3- (methoxymethoxy) naphthalen-1-yl trifluoromethanesulfonate to give the title compound (55 mg, yield: 38.3%).

MS m/z(ESI):724.4[M+H] ⁺ 。

The second step is that: preparation of 4- (4- (-8-azabicyclo [3.2.1] octan-3-yl) -2- ((2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -5, 6-dihydropyrido [3,4-d ] pyrimidin-7 (8H) -yl) -5, 6-difluoronaphthalen-2-ol (Compound 33)

Using the synthetic route of example 2, the second reaction starting material, tert-butyl 3- (7- (8-chloronaphthalen-1-yl) -2- ((2r, 7as) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidin-4-yl) -8-azabicyclo [3.2.1] octane-8-carboxylate, was replaced with tert-butyl 3- (7, 8-difluoro-3- (methoxymethoxy) naphthalen-1-yl) -2- ((2r, 7as) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidin-4-yl) -8-azabicyclo [3.2.1] octane-8-carboxylate to give the title compound (37 mg, crude).

MS m/z(ESI):580.3[M+H] ⁺ 。

The third step: 4- (4- ((1R, 3s, 5S) -8-azabicyclo [3.2.1] octan-3-yl) -2- ((2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -5, 6-dihydropyrido [3,4-d ] pyrimidin-7 (8H) -yl) -5, 6-difluoronaphthalen-2-ol (compound 33A or 33B) and

The crude product of the second step was purified by high performance liquid chromatography to give the title compound 33A (3.0 mg, yield: 4.1%) and compound 33B (1.6 mg, yield: 2.3%).

Compound 33A: the retention time was 5.15 minutes;

MS m/z(ESI):580.3[M+H] ⁺ 。

¹ H-NMR(400MHz,DMSO-d ₆ ):δ7.58-7.45(m,2H),6.91(s,1H),6.85(s,1H),5.38(s,1H),5.21(s,1H),4.14-4.04(m,4H),3.63-3.44(m,3H),3.13-2.67(m,8H),2.33-2.08(m,10H),1.90-1.80(m,5H)。

compound 33B: retention time 5.53 minutes;

MS m/z(ESI):580.3[M+H] ⁺ 。

¹ H-NMR(400MHz,DMSO-d ₆ ):δ7.58-7.45(m,2H),6.91(s,1H),6.85(s,1H),5.38(s,1H),5.21(s,1H),4.13-3.83(m,7H),3.03-2.74(m,8H),2.11-1.95(m,10H),1.89-1.78(m,5H)。

example 5: preparation of 4- ((1R, 3s, 5S) -8-azabicyclo [3.2.1] octan-3-yl) -7- (8-bromonaphthalen-1-yl) -2- ((2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidine (Compound 30A or 30B)

Preparation of 4- ((1R, 3r, 5S) -8-azabicyclo [3.2.1] octan-3-yl) -7- (8-bromonaphthalen-1-yl) -2- ((2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidine (Compound 30A or 30B)

The first step is as follows: preparation of 3- (7- (8-bromonaphthalen-1-yl) -2- ((2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidin-4-yl) -8-azabicyclo [3.2.1] octane-8-carboxylic acid tert-butyl ester

Using the synthetic route of example 2, the starting material 1-bromo-8-chloronaphthalene in the first step was replaced with 1, 8-dibromonaphthalene to give the title compound (272 mg, yield: 58.1%).

MS m/z(ESI):706.3[M+H] ⁺ 。

The second step is that: preparation of 4- (-8-azabicyclo [3.2.1] oct-3-yl) -7- (8-bromonaphthalen-1-yl) -2- ((2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidine (Compound 30)

Using the synthetic route of example 2, the second reaction starting material, tert-butyl 3- (7- (8-chloronaphthalen-1-yl) -2- ((2r, 7as) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidin-4-yl) -8-azabicyclo [3.2.1] octane-8-carboxylate, was replaced with tert-butyl 3- (7- (8-bromonaphthalen-1-yl) -2- ((2r, 7as) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidin-4-yl) -8-azabicyclo [3.2.1] octane-8-carboxylate to give the title compound (37 mg, crude).

MS m/z(ESI):580.3[M+H] ⁺ 。

The third step: 4- ((1R, 3s, 5S) -8-azabicyclo [3.2.1] octan-3-yl) -7- (8-bromonaphthalen-1-yl) -2- ((2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidine (compound 30A or 30B) and

The crude product of the second step was purified by high performance liquid chromatography to give the title compound 30A (8.2 mg, yield: 17.5%) and compound 30B (7.0 mg, yield: 15.3%).

Compound 30A: the retention time was 6.05 minutes;

MS m/z(ESI):580.3[M+H] ⁺ 。

¹ H-NMR(400MHz,DMSO-d ₆ ):δ7.97(d,J＝8.0Hz,1H),7.82(d,J＝4.0Hz,1H),7.77(d,J＝8.0Hz,1H),7.58-7.54(m,1H),7.41-7.38(m,2H),5.33(s,1H),5.20(s,1H),4.14-3.81(m,7H),3.56(d,J＝10Hz,1H),3.20-3.00(m,6H),2.82(dd,J＝12.4Hz,J＝8.4Hz,1H),2.68(d,J＝14.4Hz,1H),2.38-2.25(m,3H),2.10-1.75(m,9H)。

compound 30B: the retention time was 6.81 minutes;

MS m/z(ESI):580.3[M+H] ⁺ 。

¹ H-NMR(400MHz,DMSO-d ₆ ):δ7.97(d,J＝8.0Hz,1H),7.81(d,J＝4.0Hz,1H),7.77(d,J＝8.0Hz,1H),7.60-7.52(m,1H),7.41-7.36(m,2H),5.33(s,1H),5.21(s,1H),4.14-3.96(m,4H),3.84-3.79(m,3H),3.56(d,J＝10Hz,1H),3.23-3.00(m,6H),2.82(dd,J＝12.4Hz,J＝8.4Hz,1H),2.68(d,J＝14.4Hz,1H),2.11-1.75(m,12H)。

example 6: preparation of 4- (4- ((1R, 3s, 5S) -8-azabicyclo [3.2.1] octan-3-yl) -2- ((2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -5, 6-dihydropyrido [3,4-d ] pyrimidin-7 (8H) -yl) -5-bromonaphthalen-2-ol (compound 31A or 31B) and 4- (4- ((1R, 3r, 5S) -8-azabicyclo [3.2.1] octan-3-yl) -2- ((2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -5, 6-dihydropyrido [3,4-d ] pyrimidin-7 (8H) -yl) -5-bromonaphthalen-2-ol (compound 31A or 31B)

The first step is as follows: preparation of tert-butyl 3- (7- (8-bromo-3- (methoxymethoxy) naphthalen-1-yl) -2- ((2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidin-4-yl) -8-azabicyclo [3.2.1] octane-8-carboxylate

Using the synthetic route of example 2, the first-step reaction starting material 1-bromo-8-chloronaphthalene was replaced with 1, 8-dibromo-3- (methoxymethoxy) naphthalene to give the title compound (36 mg, yield: 22.31%).

MS m/z(ESI):766.3[M+H] ⁺ 。

The second step is that: preparation of 4- (4- (-8-azabicyclo [3.2.1] octan-3-yl) -2- ((2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -5, 6-dihydropyrido [3,4-d ] pyrimidin-7 (8H) -yl) -5-bromonaphthalen-2-ol (Compound 31)

The second reaction starting material, tert-butyl 3- (7- (8-chloronaphthalen-1-yl) -2- ((2r, 7as) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidin-4-yl) -8-azabicyclo [3.2.1] octane-8-carboxylate was replaced with tert-butyl 3- (8-bromo-3- (methoxymethoxy) naphthalen-1-yl) -2- ((2r, 7as) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidin-4-yl) -8-azabicyclo [3.2.1] octane-8-carboxylate using the synthetic route of example 2 to give the title compound (10.0 mg, crude).

MS m/z(ESI):625.3[M+H] ⁺ 。

The third step: 4- (4- ((1R, 3s, 5S) -8-azabicyclo [3.2.1] octan-3-yl) -2- ((2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -5, 6-dihydropyrido [3,4-d ] pyrimidin-7 (8H) -yl) -5-bromonaphthalen-2-ol (compound 31A or 31B) and

preparation of 4- (4- ((1R, 3r, 5S) -8-azabicyclo [3.2.1] octan-3-yl) -2- ((2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -5, 6-dihydropyrido [3,4-d ] pyrimidin-7 (8H) -yl) -5-bromonaphthalen-2-ol (Compound 31A or 31B)

The crude product of the second step was purified by high performance liquid chromatography to give the title compound 31A (3.0 mg, yield: 10.09%) and compound 31B (2.5 mg, yield: 8.41%).

Compound 31A: the retention time was 6 minutes;

MS m/z(ESI):625.3[M+H] ⁺ 。

¹ H-NMR(400MHz,DMSO-d ₆ ):δ7.70(d,J＝8.0Hz,1H),7.53(d,J＝8.0Hz,1H),7.42-7.38(m,1H),7.22-7.17(m,1H),6.98-6.91(m,1H),5.33(s,1H),5.20(s,1H),4.14-3.72(m,10H),3.03(d,J＝32.0Hz,4H),2.84(d,J＝12.0Hz,2H),2.12-1.73(m,13H)。

compound 31B: the retention time was 7.4 minutes;

MS m/z(ESI):625.3[M+H] ⁺ 。

¹ H-NMR(400MHz,DMSO-d ₆ ):δ7.69(s,1H),7.53(d,J＝8.0Hz,1H),7.22-7.17(m,1H),6.95(d,J＝16.0Hz,2H),5.33(s,1H),5.20(s,1H),4.13-3.77(m,10H),3.07(s,2H),3.00(s,2H),2.83(s,2H),2.11-1.69(m,13H)。

example 7:4- (4- (1R, 3s, 5S) -8-azabicyclo [3.2.1] octan-3-yl) -8-fluoro-2- (2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) pyridin [4,3-d ] pyrimidin-7-yl) -5-ethyl-6-fluoronaphthalen-2-ol (Compound 49A or 49B) and

preparation of 4- (4- (1R, 3r, 5S) -8-azabicyclo [3.2.1] octan-3-yl) -8-fluoro-2- (2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) pyridin [4,3-d ] pyrimidin-7-yl) -5-ethyl-6-fluoronaphthalen-2-ol (Compound 49A or 49B)

The first step is as follows: preparation of tert-butyl 3- (8-fluoro-7- (7-fluoro-3- (methoxymethyloxy) -8- (triisopropylsilyl) ethynyl) naphthalen-1-yl) -2- (2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) pyridine [4,3-d ] pyrimidin-4-yl) -8-azabicyclo [3.2.1] octane-8-carboxylate

Tert-butyl 3- (7-chloro-8-fluoro-2- ((2r, 7as) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) pyridine [4,3-d ] pyrimidin-4-yl) -8-azabicyclo [3.2.1] octane-8-carboxylate (80mg, 0.15mmol) was dissolved in 1, 4-dioxane (5 mL) and water (0.5 mL), and 2- [ 2-fluoro-6- (methoxymethoxy) -8- (4, 5-tetramethyl-1, 3, 2-dioxolan-2-yl) -1-naphthyl ] ethynyl ] triisopropylsilane (108mg, 0.21mmol), potassium phosphate (95mg, 0.45mmol) and [ n-butyldi (1-adamantyl) phosphine methanesulfonate ] (2-amino-1, 1' -biphenyl-2-yl) palladium (II) (10.9mg, 0.015 mmol) were added, and stirred for three times at 100 ℃ under nitrogen substitution for 15 hours. The reaction solution was cooled to room temperature, diluted with water (10 mL), extracted with ethyl acetate (10 mL) three times, the organic phase was washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent: dichloromethane/ethyl acetate =1/1, volume ratio) to give the title compound (61 mg, yield: 46.6%).

MS m/z(ESI):900.5[M+H] ⁺ 。

The second step is that: preparation of 3- (7- (8-ethynyl-7-fluoro-3- (methoxymethoxy) naphthalen-1-yl) -8-fluoro-2- ((2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -pyridine [4,3-d ] pyrimidin-4-yl) -8-azabicyclo [3.2.1] octane-8-carboxylic acid tert-butyl ester

Tert-butyl 3- (8-fluoro-7- (7-fluoro-3- (methoxymethoxy) -8- (triisopropylsilyl) ethynyl) naphthalen-1-yl) -2- (2r, 7as) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) pyridin [4,3-d ] pyrimidin-4-yl) -8-azabicyclo [3.2.1] octane-8-carboxylate (61mg, 67.8 μmol) was dissolved in dichloromethane (1 mL), followed by addition of tetrabutylammonium fluoride in tetrahydrofuran (1m, 27.50 μ L), nitrogen blanketed, and stirred at 25 ℃ for 0.5 hour. To the reaction solution, water (10 mL) was added, and extraction was performed twice with ethyl acetate (10 mL), and the organic phase was washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give the title compound (48 mg, yield: 95.2%).

MS m/z(ESI):744.3[M+H] ⁺ 。

The third step: preparation of 3- (7- (8-ethyl-7-fluoro-3- (methoxymethyloxy) naphthalen-1-yl) -8-fluoro-2- ((2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -pyridin [4,3-d ] pyrimidin-4-yl) -8-azabicyclo [3.2.1] octane-8-carboxylic acid tert-butyl ester

Tert-butyl 3- (7- (8-ethynyl-7-fluoro-3- (methoxymethoxy) naphthalen-1-yl) -8-fluoro-2- ((2r, 7as) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -pyridin [4,3-d ] pyrimidin-4-yl) -8-azabicyclo [3.2.1] octane-8-carboxylate (30mg, 67.8 μmol) was dissolved in methanol (1 mL), 10% palladium on carbon (5 mg) was added, hydrogen gas was introduced into the system, and stirring was carried out at 25 ℃ for 20 minutes. Filtration and concentration under reduced pressure gave the title compound (25 mg, yield: 82.8%).

MS m/z(ESI):748.4[M+H] ⁺ 。

The fourth step: preparation of 4- (4- (8-azabicyclo [3.2.1] oct-3-yl) -8-fluoro-2- (2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) pyridin [4,3-d ] pyrimidin-7-yl) -5-ethyl-6-fluoronaphthalen-2-ol (Compound 49)

Tert-butyl 3- (7- (8-ethyl-7-fluoro-3- (methoxymethoxy) naphthalen-1-yl) -8-fluoro-2- ((2r, 7as) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -pyridine [4,3-d ] pyrimidin-4-yl) -8-azabicyclo [3.2.1] octane-8-carboxylate (25mg, 33.4 μmol) was dissolved in dichloromethane (3 mL), trifluoroacetic acid (0.5 mL) was added, and the mixture was stirred at 25 ℃ for 2 hours. The reaction was concentrated, and the residue was diluted with ethyl acetate (10 mL), adjusted to pH =8 to 9 with sodium bicarbonate solution, separated, and the organic phase was dried over anhydrous sodium sulfate and concentrated to give the title compound (15 mg, crude).

MS m/z(ESI):604.3[M+H] ⁺ 。

The fifth step: 4- (4- (1R, 3s, 5S) -8-azabicyclo [3.2.1] octan-3-yl) -8-fluoro-2- (2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) pyridin [4,3-d ] pyrimidin-7-yl) -5-ethyl-6-fluoronaphthalen-2-ol (compound 49A or 49B) and

The crude product of the fourth step was purified by high performance liquid chromatography to give the title compound 49A (1.0 mg, yield: 5%) and compound 49B (1.5 mg, yield: 7.5%).

Compound 49A: the retention time was 5.6 minutes;

MS m/z(ESI):604.3[M+H] ⁺ 。

¹ H-NMR(400MHz,DMSO-d ₆ ):δ9.63(s,1H),8.30(s,1H),7.80(d,J＝4.0Hz,1H),7.37(d,J＝4.0Hz,2H),7.03(s,1H),5.91(d,J＝4.0Hz,1H),5.86(d,J＝4.0Hz,1H),5.37(s,1H),5.23(s,1H),4.26-4.16(m,3H),3.96-3.90(m,2H),3.09-3.02(m,2H),2.89-2.83(m,1H),2.69-2.66(m,4H),2.34-2.29(m,4H),2.20-2.13(m,3H),2.10-2.05(m,2H),1.95-1.83(m,5H)。

compound 49B: the retention time was 6.0 minutes;

MS m/z(ESI):604.3[M+H] ⁺ 。

¹ H-NMR(400MHz,DMSO-d ₆ ):δ9.63(s,1H),8.30(s,1H),7.80(d,J＝4.0Hz,1H),7.37(d,J＝4.0Hz,2H),7.03(s,1H),5.91(d,J＝4.0Hz,1H),5.86(d,J＝4.0Hz,1H),5.35-5.28(m,2H),4.22-4.13(m,3H),3.99-3.90(m,2H),3.12-3.05(m,2H),2.88-2.84(m,1H),2.68-2.64(m,4H),2.34-2.29(m,4H),2.20-2.13(m,3H),2.13-2.06(m,2H),1.99-1.87(m,5H)。

example 8: preparation of 4- (4- (8-azabicyclo [3.2.1] octan-3-yl) -8-fluoro-2- (2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) pyridin [4,3-d ] pyrimidin-7-yl) -5-ethynyl-6-fluoronaphthalen-2-ol (Compound 44)

Tert-butyl 3- (7- (8-ethyl-7-fluoro-3- (methoxymethoxy) naphthalen-1-yl) -8-fluoro-2- ((2r, 7as) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -pyridine [4,3-d ] pyrimidin-4-yl) -8-azabicyclo [3.2.1] octane-8-carboxylate (18mg, 24.1 μmol) was dissolved in dichloromethane (2 mL), trifluoroacetic acid (0.4 mL) was added, and the mixture was stirred at 25 ℃ for 2 hours. The reaction solution was concentrated, and the residue was diluted with ethyl acetate (10 mL), adjusted to pH =8 to 9 with sodium bicarbonate solution, separated, the organic phase was dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by high performance liquid chromatography to give the title compound (1.2 mg, yield: 8.3%).

MS m/z(ESI):600.3[M+H] ⁺ 。

¹ H-NMR(400MHz,DMSO-d ₆ ):δ9.61(s,1H),8.00(dd,J＝8.0Hz,J＝4.0Hz,1H),7.51-7.43(m,2H),7.20(s,1H),5.91(d,J＝4.0Hz,1H),5.86-5.81(m,1H),5.37-5.31(m,1H),4.31-4.13(m,4H),4.06-4.00(m,2H),3.80(s,1H),3.10-3.04(m,2H),2.89-2.83(m,1H),2.68(t,J＝4.0Hz,2H),2.37-2.28(m,4H),2.16-2.09(m,2H),2.03-1.97(m,4H),1.87-1.81(m,2H)。

Example 9:4- (4- ((1R, 3s, 5S) -8-azabicyclo [3.2.1] octan-3-yl) -2- ((2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -5, 6-dihydropyridin [3,4-d ] pyrimidin-7 (8H) -yl) -5-chloronaphthalen-2-ol (compound 36A or 36B) and

preparation of 4- (4- ((1R, 3r, 5S) -8-azabicyclo [3.2.1] octan-3-yl) -2- ((2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -5, 6-dihydropyridin [3,4-d ] pyrimidin-7 (8H) -yl) -5-chloronaphthalen-2-ol (Compound 36A or 36B)

The first step is as follows: preparation of tert-butyl 3- (7- (8-chloro-3- (methoxymethyloxy) naphthalen-1-yl) -2- ((2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -5,6,7, 8-tetrahydropyridine [3,4-d ] pyrimidin-4-yl) -8-azabicyclo [3.2.1] octane-8-carboxylate

Using the synthetic route of example 2, the first reaction starting material, 1-bromo-8-chloronaphthalene, was replaced with 1-bromo-8-chloro-3- (methoxymethoxy) naphthalene to give the title compound (59 mg, yield: 68.3%).

MS m/z(ESI):722.3[M+H] ⁺ 。

The second step is that: preparation of 4- (4- ((1R, 5S) -8-azabicyclo [3.2.1] octan-3-yl) -2- ((2R, 7aS) -2-fluorohexahydro-1H-pyrrolin-7 a-yl) methoxy) -5, 6-dihydropyridin [3,4-d ] pyrimidin-7 (8H) -yl) -5-chloronaphthalen-2-ol (Compound 36)

Using the synthetic route of example 2, the second reaction starting material, tert-butyl 3- (7- (8-chloronaphthalen-1-yl) -2- ((2r, 7as) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidin-4-yl) -8-azabicyclo [3.2.1] octane-8-carboxylate, was replaced with tert-butyl 3- (7- (8-chloro-3- (methoxymethoxy) naphthalen-1-yl) -2- ((2r, 7as) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -5,6,7, 8-tetrahydropyridin [3,4-d ] pyrimidin-4-yl) -8-azabicyclo [3.2.1] octane-8-carboxylate to give the title compound (10.0 mg, crude).

MS m/z(ESI):578.3[M+H] ⁺ 。

The third step: 4- (4- ((1R, 3s, 5S) -8-azabicyclo [3.2.1] octan-3-yl) -2- ((2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -5, 6-dihydropyridin [3,4-d ] pyrimidin-7 (8H) -yl) -5-chloronaphthalen-2-ol (compound 36A or 36B) and

The crude product of the second step was purified by high performance liquid chromatography to give the title compound 36A (3.0 mg, yield: 10.71%) and compound 36B (2.5 mg, yield: 8.93%).

Compound 36A: retention time 6.35 minutes;

MS m/z(ESI):578.3[M+H] ⁺ 。

¹ H-NMR(400MHz,DMSO-d ₆ ):δ7.66(dt,J＝8.0Hz,J＝4.0Hz,1H),7.29(dt,J＝8.0Hz,J＝4.0Hz,2H),6.96(d,J＝4.0Hz,1H),6.88(d,J＝4.0Hz,1H),5.34(s,1H),5.20(s,1H),4.17-3.93(m,7H),3.83(s,3H),3.01(s,4H),2.82(d,J＝8.0Hz,2H),1.97(dt,J＝20.0Hz,J＝16.0Hz,7H),1.81(d,J＝16.0Hz,6H)。

compound 36B: retention time was 6.7 minutes;

MS m/z(ESI):578.3[M+H] ⁺ 。

¹ H-NMR(400MHz,DMSO-d ₆ ):δ7.66(dt,J＝8.0Hz,J＝4.0Hz,1H),7.30-7.28(m,2H),6.96(d,J＝4.0Hz,1H),6.89(s,1H),5.34(s,1H),5.20(s,1H),4.14(d,J＝16.0Hz,2H),4.05-3.99(m,2H),3.94-3.82(m,6H),3.07(s,4H),2.82(d,J＝8.0Hz,2H),2.11(s,2H),2.03(s,2H),1.96(s,3H),1.83(d,J＝20.0Hz,3H),1.73(d,J＝12.0Hz,3H)。

example 10:4- (4- (1R, 3s, 5S) -8-azabicyclo [3.2.1] octan-3-yl) -2- (2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7A-yl) methoxy) -5, 6-dihydropyrido [3,4-d ] pyrimidin-7 (8H) -yl) -5-ethylnaphthalen-2-ol (compound 37A or compound 37B) and

preparation of 4- (4- (1R, 3r, 5S) -8-azabicyclo [3.2.1] octan-3-yl) -2- (2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7A-yl) methoxy) -5, 6-dihydropyridin [3,4-d ] pyrimidin-7 (8H) -yl) -5-ethylnaphthalen-2-ol (Compound 37A or Compound 37B)

The first step is as follows: preparation of 3- (2- (2R, 7aS) -2-Fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -7- (3- (methoxymethoxy) -8- (triisopropylsilyl) ethynyl) naphthalen-1-yl) -5,6,7, 8-tetrahydropyridine [3,4-d ] pyrimidin-4-yl) -8-azabicyclo [3.2.1] octane-8-carboxylic acid tert-butyl ester

Tert-butyl 3- (7- (8-bromo-3- (methoxymethoxy) naphthalen-1-yl) -2- ((2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidin-4-yl) -8-azabicyclo [3.2.1] octane-8-carboxylate (150mg, 195.63mmol) was dissolved in N, N-dimethylacetamide (6 mL), followed by addition of copper iodide (6.84mg, 35.91. Mu. Mol), bistriphenylphosphine palladium dichloride (25.22mg, 35.93. Mu. Mol), N-diisopropylethylamine (229.85mg, 1.79mmol), nitrogen substitution, and reaction at 100 ℃ for 1.5 hours. After the system was cooled to room temperature, 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 (110 mg, yield: 64.8%).

MS m/z(ESI):868.5[M+H] ⁺ 。

The second step: preparation of tert-butyl 3- (7- (8-ethynyl-3- (methoxymethyloxy) naphthalen-1-yl) -2- (2R, 7aS) -2-fluorohexahydro-1H-pyrrolidin-7 a-yl) methoxy) -5,6,7, 8-tetrahydropyridine [3,4-d ] pyrimidin-4-yl) -8-azabicyclo [3.2.1] octane-8-carboxylate

Tert-butyl 3- (2- (2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -7- (3- (methoxymethoxy) -8- (triisopropylsilyl) ethynyl) naphthalen-1-yl) -5,6,7, 8-tetrahydropyridin [3,4-d ] pyrimidin-4-yl) -8-azabicyclo [3.2.1] octane-8-carboxylate (110mg, 126.7mmol) was dissolved in tetrahydrofuran (2 mL), the temperature was lowered to 0 ℃ for replacement with nitrogen, a tetrahydrofuran solution (1M, 1mL) of tetrabutylammonium fluoride was added, and the reaction was carried out at room temperature for 0.5 hour. The reaction solution was diluted with ethyl acetate (20 mL), washed twice with water (40 mL), and the organic phase was collected, dried over anhydrous sodium sulfate, and concentrated to give the title compound (80 mg, yield: 88.8%).

MS m/z(ESI):712.4[M+H] ⁺ 。

The third step: preparation of 3- (7- (8-Ethyl-3- (methoxymethyloxy) naphthalen-1-yl) -2- (2R, 7aS) -2-Fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -5,6,7, 8-tetrahydropyridine [3,4-d ] pyrimidin-4-yl) -8-azabicyclo [3.2.1] octane-8-carboxylic acid tert-butyl ester

Tert-butyl 3- (7- (8-ethynyl-3- (methoxymethoxy) naphthalen-1-yl) -2- (2R, 7aS) -2-fluorohexahydro-1H-pyrrolidin-7 a-yl) methoxy) -5,6,7, 8-tetrahydropyridine [3,4-d ] pyrimidin-4-yl) -8-azabicyclo [3.2.1] octane-8-carboxylate (60mg, 84.3. Mu. Mol) was dissolved in anhydrous methanol (4 mL), 10% palladium on carbon (29 mg) was added thereto, hydrogen substitution was carried out three times, and the reaction was carried out at 25 ℃ for 18 hours. The reaction solution was filtered, the filter cake was washed with methanol (4 mL), the filtrate was concentrated, and the residue was purified by silica gel column chromatography (eluent: dichloromethane/methanol =10/1, volume ratio) to obtain the title compound (19 mg, yield: 31.5%).

MS m/z(ESI):716.4[M+H] ⁺ 。

The fourth step: preparation of 4- (4- (1R, 5S) -8-azabicyclo [3.2.1] octan-3-yl) -2- (2R, 7aS) -2-fluorohexahydro-1H-pyrrolin-7 a-yl) methoxy) -5, 6-dihydropyrido [3,4-d ] pyrimidin-7 (8H) -yl) -5-ethylnaphthalen-2-ol (Compound 37)

(1R, 5S) -tert-butyl 3- (7- (8-ethyl-3- (methoxymethyloxy) naphthalen-1-yl) -2- (2R, 7aS) -2-fluorohexahydro-1H-pyrrolin-7 a-yl) methoxy) -5,6,7, 8-tetrahydropyridin [3,4-d ] pyrimidin-4-yl) -8-azabicyclo [3.2.1] octane-8-carboxylic acid tert-butyl ester (19mg, 26.5. Mu. Mol) was dissolved in dichloromethane (2 mL), trifluoroacetic acid (1 mL) was added dropwise at 25 ℃ and the reaction was stirred at 25 ℃ for 1.5 hours. The reaction was concentrated, the residue was diluted with ethyl acetate (3 mL), washed three times with sodium bicarbonate solution (10 mL), the organic phases were combined, back-washed twice with saturated brine (10 mL), dried over anhydrous sodium sulfate, and the organic solvent was concentrated to give the title compound (13 mg, crude).

The fifth step: 4- (4- (1R, 3s, 5S) -8-azabicyclo [3.2.1] octan-3-yl) -2- (2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7A-yl) methoxy) -5, 6-dihydropyrido [3,4-d ] pyrimidin-7 (8H) -yl) -5-ethylnaphthalen-2-ol (compound 37A or compound 37B) and

The crude product of the fourth step was purified by high performance liquid chromatography to give the title compound 37A (1.0 mg, yield: 6.6%) and compound 37B (2.0 mg, yield: 13.2%).

Compound 37A: the retention time was 7.45 minutes;

MS m/z(ESI):572.3[M+H] ⁺ 。

¹ H NMR(400MHz,CD ₃ OD)δ7.49(d,J＝8.0Hz,1H),7.25-7.21(m,1H),7.05(d,J＝8.0Hz,1H),6.95(d,J＝2.0Hz,1H),6.91(d,J＝2.0Hz,1H),5.45(s,1H),5.32(s,1H),4.34(dd,J＝12.0Hz,J＝8.0Hz,1H),4.32-4.27(m,1H),4.17-4.09(m,3H),3.83(d,J＝16.0Hz,1H),3.62-3.51(m,3H),3.48(dd,J＝8.0Hz,J＝4.0Hz,2H),3.41(s,1H),3.20-3.11(m,2H),3.08-2.97(m,3H),2.79(s,1H),2.53-2.47(m,2H),2.37-2.32(m,2H),2.22(dd,J＝16.0Hz,J＝4.0Hz,4H),2.13-2.05(m,4H),1.29(d,J＝8.0Hz,2H),1.06(d,J＝8.0Hz,2H)。

compound 37B: the retention time was 7.85 minutes;

MS m/z(ESI):572.3[M+H] ⁺ 。

¹ H NMR(400MHz,CD ₃ OD)δ7.48(d,J＝8.0Hz,1H),7.25-7.21(m,1H),7.06-7.02(m,1H),6.95(d,J＝2.0Hz,1H),6.92(s,1H),5.42(s,1H),5.29(s,1H),4.28(t,J＝12.0Hz,2H),4.16(s,2H),4.10(s,1H),3.82(d,J＝16.0Hz,1H),3.57(d,J＝8.8Hz,3H),3.42(s,2H),3.35(s,1H),3.12(s,2H),3.05-2.93(m,3H),2.32(d,J＝16.0Hz,4H),2.21(s,4H),2.07(d,J＝4.0Hz,3H),1.92(d,J＝12.0Hz,3H),1.29(t,J＝8.0Hz,1H),1.05(d,J＝8.0Hz,2H)。

example 11:4- ((1R, 3s, 5S) -8-azabicyclo [3.2.1] octan-3-yl) -7- (8-chloro-7-fluoronaphthalen-1-yl) -2- (((2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidine (compound 39A or 39B) and

preparation of 4- ((1R, 3r, 5S) -8-azabicyclo [3.2.1] octan-3-yl) -7- (8-chloro-7-fluoronaphthalen-1-yl) -2- (((2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidine (Compound 39A or 39B)

The first step is as follows: preparation of tert-butyl 3- (7- (8-chloro-7-fluoronaphthalen-1-yl) -2- (((2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidin-4-yl) -8-azabicyclo [3.2.1] octane-8-carboxylate

Using the synthetic route of example 2, the first step reaction starting material 1-bromo-8-chloronaphthalene was replaced with 8-chloro-7-fluoronaphthalen-1-yl trifluoromethanesulfonate to give the title compound (150 mg, yield: 73.75%).

MS m/z(ESI):680.3[M+H] ⁺ 。

The second step is that: preparation of 4- (8-azabicyclo [3.2.1] octan-3-yl) -7- (8-chloro-7-fluoronaphthalen-1-yl) -2- (((2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidine (Compound 39)

Using the synthetic route of example 2, the second reaction starting material, tert-butyl 3- (7- (8-chloronaphthalen-1-yl) -2- ((2r, 7as) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidin-4-yl) -8-azabicyclo [3.2.1] octane-8-carboxylate, was replaced with tert-butyl 3- (7- (8-chloro-7-fluoronaphthalen-1-yl) -2- (((2r, 7as) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidin-4-yl) -8-azabicyclo [3.2.1] octane-8-carboxylate to give the title compound (50.0 mg, crude).

MS m/z(ESI):580.2[M+H] ⁺ 。

The third step: 4- ((1R, 3s, 5S) -8-azabicyclo [3.2.1] octan-3-yl) -7- (8-chloro-7-fluoronaphthalen-1-yl) -2- (((2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidine (compound 39A or 39B) and

The crude product of the second step was purified by high performance liquid chromatography to give the title compound 39A (18.0 mg, yield: 20.71%) and compound 39B (18.0 mg, yield: 20.71%).

Compound 1A: retention time was 5.7 minutes;

MS m/z(ESI):580.2[M+H] ⁺ 。

¹ H-NMR(400MHz,DMSO-d ₆ ):δ8.02(dd,J＝8.0Hz,J＝4.0Hz,1H),7.80(d,J＝8.0Hz,1H),7.62-7.52(m,2H),7.44(d,J＝8.0Hz,1H),5.27(d,J＝4.0Hz,1H),4.21-3.81(m,7H),3.61-3.30(m,3H),3.22-2.95(m,5H),2.86-2.63(m,2H),2.15-1.67(m,13H)。

compound 1B: the retention time was 6.3 minutes;

MS m/z(ESI):580.2[M+H] ⁺ 。

¹ H-NMR(400MHz,DMSO-d ₆ ):δ8.03(dd,J＝8.0Hz,J＝4.0Hz,1H),7.80(d,J＝8.0Hz,1H),7.62-7.52(m,2H),7.44(d,J＝8.0Hz,1H),5.27(d,J＝4.0Hz,1H),4.15-3.92(m,6H),3.54-2.81(m,9H),2.11-1.70(m,15H)。

example 12:4- ((1R, 3s, 5S) -8-azabicyclo [3.2.1] octan-3-yl) -7- (8-ethyl-7-fluoronaphthalen-1-yl) -2- (((2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidine (compound 40A or 40B) and

preparation of 4- ((1R, 3r, 5S) -8-azabicyclo [3.2.1] octan-3-yl) -7- (8-ethyl-7-fluoronaphthalen-1-yl) -2- (((2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidine (Compound 40A or 40B)

The first step is as follows: preparation of tert-butyl 3- (7- (7-fluoro-8-vinylnaphthalen-1-yl) -2- (((2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidin-4-yl) -8-azabicyclo [3.2.1] octane-8-carboxylate

3- (7- (8-chloro-7-fluoronaphthalen-1-yl) -2- (((2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidin-4-yl) -8-azabicyclo [3.2.1] octane-8-carboxylic acid tert-butyl ester (120mg, 176.41. Mu. Mol), potassium ethylenetrifluoroborate (70.99mg, 0.53mmol), cesium fluoride (106.33mg, 0.7mmol), tetrakis (triphenylphosphine) palladium (20.38mg, 17.64. Mu. Mol) were sequentially added to 1, 4-dioxane (3 mL) and water (0.5 mL), replaced three times with nitrogen gas, and the reaction system was microwave-reacted at 100 ℃ for 3 hours. After the system was cooled to room temperature, the solvent was rotovapped off under reduced pressure, and the residue was purified by silica gel column chromatography (eluent: dichloromethane/methanol =10/1, volume ratio) to obtain the title compound (100 mg, yield: 84.37%).

MS m/z(ESI):672.3[M+H] ⁺ 。

The second step is that: preparation of tert-butyl 3- (7- (8-ethyl-7-fluoronaphthalen-1-yl) -2- (((2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidin-4-yl) -8-azabicyclo [3.2.1] octane-8-carboxylate

Tert-butyl 3- (7- (7-fluoro-8-vinylnaphthalen-1-yl) -2- (((2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidin-4-yl) -8-azabicyclo [3.2.1] octane-8-carboxylate (100mg, 0.15mmol) was added to methanol (2 mL), 10% palladium on carbon (60 mg) was added thereto, the mixture was replaced with hydrogen gas three times, and the mixture was stirred at 20 ℃ for 12 hours. The reaction solution was filtered, and concentrated to give the title compound (50 mg, yield: 49.89%).

MS m/z(ESI):674.3[M+H] ⁺ 。

The third step: preparation of 4- (8-azabicyclo [3.2.1] octan-3-yl) -7- (8-ethyl-7-fluoronaphthalen-1-yl) -2- (((2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidine

3- (7- (8-Ethyl-7-fluoronaphthalen-1-yl) -2- (((2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidin-4-yl) -8-azabicyclo [3.2.1] octane-8-carboxylic acid tert-butyl ester (50.0 mg, 87.15. Mu. Mol) was dissolved in dichloromethane (3 mL), trifluoroacetic acid (0.5 mL) was added dropwise at 25 ℃, and the reaction was stirred at 25 ℃ for 1.5 hours. The reaction was concentrated, the residue was diluted with ethyl acetate (3 mL), washed three times with sodium bicarbonate solution (10 mL), backwashed twice with saturated brine (10 mL), dried over anhydrous sodium sulfate, and the solvent was rotary evaporated under reduced pressure to give the title compound (35.0 mg, crude).

MS m/z(ESI):574.3[M+H] ⁺ 。

The fourth step: 4- ((1R, 3s, 5S) -8-azabicyclo [3.2.1] octan-3-yl) -7- (8-ethyl-7-fluoronaphthalen-1-yl) -2- (((2R, 7aS) -2-fluorohexahydro-1H-pyrrolizin-7 a-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3,4-d ] pyrimidine (compound 40A or 40B) and

The crude product of the third step was purified by high performance liquid chromatography to give the title compound 40A (5.0 mg, yield: 11.74%) and compound 40B (5.0 mg, yield: 11.74%).

Compound 2A: the retention time was 6.1 minutes;

MS m/z(ESI):574.3[M+H] ⁺ 。

¹ H-NMR(400MHz,DMSO-d ₆ ):δ7.87(dd,J＝8Hz,J＝1.2Hz,1H),7.78(d,J＝8.0Hz,1H),7.52-7.46(m,2H),7.39(t,J＝8.0Hz,1H),5.26(d,J＝12.0Hz,1H),3.99-3.91(m,6H),3.30(d,J＝8.0Hz,4H),3.06-2.99(m,5H),2.81(dd,J＝16.0Hz,J＝8.0Hz,2H),2.44(s,2H),2.09-1.75(m,13H),1.02(t,J＝8.0Hz,3H)。

compound 2B: the retention time was 7.0 minutes;

MS m/z(ESI):574.3[M+H] ⁺ 。

¹ H-NMR(400MHz,DMSO-d ₆ ):δ7.87(dd,J＝8.0Hz,J＝1.2Hz,1H),7.78(dd,J＝8.0Hz,J＝4.0Hz,1H),7.53-7.46(m,2H),7.39(t,J＝8.0Hz,1H),5.26(d,J＝12.0Hz,1H),4.04-3.85(m,6H),3.46(s,3H),3.06-2.95(m,6H),2.82-2.80(m,2H),2.09-1.66(m,15H),1.01(t,J＝8.0Hz,3H)。

biological assay

Experimental example 1: KRAS-G12D (guanine-5' -triphosphate (GTP) hydrolase G12D mutant)/SOS 1 in vitro protein binding inhibition Activity assay

An experimental system:

KRAS-G12D/SOS1 binding assay kit (manufacturer: cisbio)

Protein: tag1-SOS1 and Tag2-KRAS G12D

Labeling: anti tag1 tb ³⁺ And Anti tag2XL665

Buffer solution: diluent Buffer and Detection Buffer

Activating nucleotide: GTP (manufacturer Sigma)

Experimental parameters:

KRAS-G12D/SOS1 binding assay kit component: tag1-SOS1, tag2-KRAS G12D, anti Tag1 tb ³⁺ And Anti tag2XL665 are both 1x

GTP：10000nM

Preincubation binding time of compound and KRAS-G12D/SOS1 protein: room temperature for 15 minutes

Protein and tag reaction time: sealing and incubating at 4 deg.C for 3 hr or extending overnight

Parameters of the microplate reader: BMG PHERAStar Fluorescence, homogeneous time-resolved Fluorescence (HTRF) method, excitation wavelength 337nm, emission wavelengths 620nm and 665nm

The experimental steps are as follows:

and (3) incubating the compound to be tested with a mixture of Tag1-SOS1 and Tag2-KRAS G12D and GTP for 15 minutes at room temperature under a Diluent Buffer system for protein binding. Adding a label diluted by Detection Buffer into the reaction plate, incubating for 3 hours at 4 ℃ or prolonging to overnight, putting the reaction plate into a microplate reader, and reading the signal value of each hole in the plate by adopting an HTRF method.

Data processing:

using a solvent group (containing 1xTag1-SOS1, 1xTag2-KRAS G12D, anti tag1 tb ³⁺ And Anti Tag2XL665, 1% DMSO) as negative control, and a reaction buffer group (containing 1xTag1-SOS1 and Anti Tag1 tb) ³⁺ And Anti tag2XL665, 1% dmso) as blanks, the relative inhibitory activity was calculated for each concentration group, with an inhibition =100% - (test group-blank group)/(vehicle group-blank group) × 100%. Half maximal Inhibitory Concentration (IC) of the compound was calculated according to a four parameter model fitting curve ₅₀ )。

The experimental results are as follows:

the inhibitory activity of the compounds on KRAS-G12D/SOS1 protein binding in vitro was determined as described above and the results are shown in Table 1.

TABLE 1 KRAS-G12D/SOS1 in vitro enzymology binding inhibition Activity test results

And (4) conclusion:

in KRAS-G12D/SOS1 in-vitro protein binding inhibition activity test, the compound of the invention shows stronger inhibition activity.

Experimental example 2: surface Plasmon Resonance (SPR) binding experiments of compounds and His-KRAS (G12D & Q61H) proteins

An experimental system:

protein: his-KRAS (G12D & Q61H)

Experimental parameters:

His-KRAS (G12D & Q61H) concentration: 0.04mg/mL

And (3) buffer solution system:

1) Protein fixation buffer 10mM HEPES,150mM NaCl,0.05% Tween-20and 10. Mu.M GDP

2) Buffer A:10mM HEPES,150mM NaCl,0.05% Tween-20and 10. Mu.M GDP

3) Buffer B10 mM HEPES,150mM NaCl,0.05% Tween-20and 10. Mu.M GDP,1% DMSO

Compound concentration: 50-0.781 mu M, two-fold gradient dilution, DMSO final concentration of 1%

Binding dissociation time: 120s on/200s off

Flow rate: 50 mu L/min

The instrument parameters are as follows: biacore T200 (GE Healthcare)

Chip: s series NTA sensor Chip

The experimental steps are as follows:

(1) Protein fixation

1) Temperature setting: the instrument temperature was set to 25 ℃.

2) The S-series NTA sensor chip is inserted, and then the "Prime" command is executed.

3) The manual program was started and the flow rate was set to 10uL/min.

4) The flow cell 2 was selected, and then a 350mM EDTA (pH 8.0) solution and a 50mM NaOH solution (the injection rate was 60. Mu.L/min, the injection time was 60 seconds, each was repeated twice) were automatically injected in sequence to wash the surface of the chip.

5) Chip surface activation: flow cell 2 was selected and 10mM NiCl2 was injected automatically at a flow rate of 4. Mu.L/min for 500s.

6) His-KRAS (G12D & Q61H) was diluted to 40. Mu.g/mL with protein immobilization buffer 120. UL of His-KRAS (G12D & Q61H) (injection rate 4. Mu.L/min, injection time set to 1150 s) was automatically injected to immobilize His-KRAS (G12D & Q61H) protein on the surface of the flow cell 2.

7) The flow rate was adjusted back to 10uL/min, and the chip surface was equilibrated to baseline essentially steady (baseline drop rate <2 RU/min).

(2) Sample detection

The program is written in Biacore T200 control software using the method schema. The temperature was set at 25 ℃ and the flow rate was set at 50uL/min. The procedure was started with 8 injections of buffer B, followed by automatic injections of compounds sequentially set from small to large concentrations, which flowed across the immobilized protein surface on the chip to bind to the chip. Flow cell 1 was set as a reference channel and 0 concentration was set as a built-in blank. Each cycle used a 50% dmso wash needle. The signal effect of DMSO was corrected with a 1% DMSO standard curve.

(3) Data processing:

binding and dissociation signals of compounds and proteins were recorded in real time and signal values for reference channels and built-in blanks were subtracted at data processing (Biacore T200 Evaluation Software) (double subtraction). The signal values for the reference channel and the built-in blank were double subtracted sensorgrams fitted with kinetic or steady state Affinity (1. By K _D Value (K) _d /K _a ) Characterizing the affinity of a compound for a protein, wherein K _d As dissociation constant, K _a To be combined withAnd (4) counting.

The experimental results are as follows:

the affinity of the compounds for His-KRAS (G12D & Q61H) protein was determined as described above and is shown in Table 2:

TABLE 2 affinity test results of compounds with His-KRAS (G12D & Q61H)

Compound (I)	K _D (nM)
		36A	11
37A	125
		39A	90.7
49B	401

And (4) conclusion:

the compounds of the present invention showed strong affinity in the binding assay of the compounds to His-KRAS (G12D & Q61H).

The above examples do not limit the scheme of the present application 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 (including all patents, patent applications, journal articles, books, and any other publications) cited in this application is hereby incorporated by reference in its entirety.

Claims

1. A compound of formula (I) or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound or metabolite thereof:

wherein,

ring A is 5-7 membered heterocycle, 5-6 membered heteroaromatic ring, C _5-7 Cycloalkyl, phenyl ring, 8-11 membered spiroheterocycle or C _8-11 Spirocycloalkyl, the A ring being optionally substituted by one or more R ³ Substitution;

X ¹ selected from N and CR ⁴ ；

L ² Selected from the group consisting of covalent bonds, -O-, and-C (O) -;

R ⁵ selected from hydrogen and C _1-6 An alkyl group;

R ⁷ Selected from hydrogen, halogen, hydroxy, cyano, C _1-6 Alkyl radical, C _2-6 Alkenyl radical, C _2-6 Alkynyl, -O-C _1-6 Alkyl, -O-C _3-6 Cycloalkyl, C _3-10 Cycloalkyl, 3-12 membered heterocyclyl and 5-10 membered heteroaryl, each optionally substituted with one or more substituents selected from hydrogen, halogen, hydroxy, cyano, C _1-6 Alkyl and C _1-6 Substituted with a haloalkyl; or,

R ⁹ selected from hydrogen, C _1-6 Alkyl radical, C _1-6 Haloalkyl, -C _1-6 alkylene-O-C _1-6 Alkyl radical, C _3-10 Cycloalkyl, 3-12 membered heterocyclyl, each of said cycloalkyl or heterocyclyl being optionally substituted with one or more substituents selected from hydrogen, halogen, hydroxy, cyanoBase, C _1-6 Alkyl radical, C _1-6 Substituted with a haloalkyl;

R ¹⁰ selected from hydrogen and C _1-6 An alkyl group;

R ¹¹ selected from hydrogen, C _1-6 Alkyl radical, C _1-6 Haloalkyl, C _3-10 Cycloalkyl and 3-12 membered heterocyclyl, said cycloalkyl or heterocyclyl being optionally substituted with one or more substituents selected from hydrogen, halogen, hydroxy, cyano, C _1-6 Alkyl radical, C _1-6 Substituted with a haloalkyl;

R ¹² selected from hydrogen and C _1-6 An alkyl group;

n is 0, 1 or 2;

provided that when

Is->

Is->

-L ¹ -R ¹ Is composed of

When is-L ² -R ² Is not->

2. The compound of claim 1, or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, or metabolite thereof, wherein:

X ¹ selected from N and CR ⁴ ，R ⁴ Selected from hydrogen, halogen, cyano, C _1-4 Alkyl andC _1-4 a haloalkyl group;

preferably, X ¹ Selected from the group consisting of N, CF, C-CN and CH;

more preferably, X ¹ Selected from N and CH;

more preferably, X ¹ Is N.

3. The compound of claim 1 or 2, or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, or metabolite thereof, wherein:

L ¹ selected from covalent bonds, -O-, -C (O) -, and-S-, -NH-and-N (C) _1-3 Alkyl) -;

preferably, L ¹ Selected from-O-, -NH-and-N (C) _1-3 Alkyl) -;

more preferably, L ¹ is-O-.

4. The compound of any one of claims 1-3, or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, or metabolite thereof, wherein:

L ² selected from the group consisting of a covalent bond and-C (O) -;

preferably, L ² Is a covalent bond.

5. The compound of any one of claims 1-4, or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, or metabolite thereof, wherein:

R ¹ is selected from C _1-6 Alkyl radical, C _3-6 Cycloalkyl and 3-8 membered heterocyclyl, each of which is optionally substituted with one or more R ⁶ Substitution; each R ⁶ Each independently selected from hydrogen, halogen, cyano, -OR ⁹ 、-O-C(O)-N(R ⁹ )R ¹⁰ 、-O-C(O)-R ⁹ 、-N(R ⁹ )R ¹⁰ 、-N(R ¹⁰ )-C(O)-R ⁹ 、-N(R ¹⁰ )-C(O)-N(R ⁹ )R ¹⁰ 、-N(R ¹⁰ )-C(O)-OR ⁹ 、-C(O)-R ⁹ 、-C(O)-OR ¹⁰ 、-C(O)-N(R ⁹ )R ¹⁰ 、C _1-6 Alkyl radical, C _1-6 Haloalkyl, C _3-10 Cycloalkyl radical, C _6-10 Aryl, 5-10 membered heteroaryl, and 3-12 membered heterocyclyl, each of which is optionally substituted with one or more substituents independently selected from the group consisting of: hydrogen, halogen, hydroxy, amino, cyano, C _1-6 Alkyl radical, C _1-6 Haloalkyl, -C _1-6 alkylene-R ¹¹ 、-C _1-6 alkylene-OR ¹¹ and-C _1-6 alkylene-NR ¹¹ R ¹² ；

Preferably, R ¹ Is selected from C _1-4 Alkyl radical, C _3-6 Cycloalkyl and 3-8 membered heterocyclyl, each of said alkyl, cycloalkyl, and heterocyclyl is optionally substituted with one or more R ⁶ Substituted, each R ⁶ Each independently selected from-OR ⁹ 、-O-C(O)-N(R ⁹ )R ¹⁰ 、-N(R ⁹ )R ¹⁰ 、-N(R ¹⁰ )-C(O)-R ⁹ 、-N(R ¹⁰ )-C(O)-OR ⁹ 、-C(O)-N(R ⁹ )R ¹⁰ 、C _3-10 Cycloalkyl, 5-10 membered heteroaryl, and 3-12 membered heterocyclyl, each of which is optionally substituted with one or more substituents independently selected from the group consisting of: hydrogen, halogen, hydroxy, amino, cyano, C _1-6 Alkyl, -C _1-6 alkylene-R ¹¹ 、-C _1-6 alkylene-OR ¹¹ and-C _1-6 alkylene-NR ¹¹ R ¹² ；

R ¹⁰ selected from hydrogen and C _1-6 An alkyl group;

R ¹² selected from hydrogen and C _1-6 An alkyl group;

preferably, R ¹ Is C _1-4 An alkyl group optionally substituted with one or more R ⁶ Substituted, each R ⁶ Each independently selected from C _3-10 Cycloalkyl, 5-10 membered heteroaryl, and 3-12 membered heterocyclyl, each of which is optionally substituted with one or more substituents independently selected from the group consisting of: halogen, C _1-6 Alkyl, -C _1-6 alkylene-R ¹¹ and-C _1-6 alkylene-NR ¹¹ R ¹² ；

R ¹¹ Selected from hydrogen, C _1-6 Alkyl and 3-12 membered heterocyclyl;

R ¹² is C _1-6 An alkyl group.

6. The compound of any one of claims 1-5, or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, or metabolite thereof, wherein:

R ² is selected from C _6-10 Aryl and 5-10 membered heteroaryl, each optionally substituted with one or more R ⁷ Substitution; each R ⁷ Each independently selected from hydrogen, halogen, hydroxy, cyano, C _1-3 Alkyl radical, C _2-4 Alkenyl radical, C _2-4 Alkynyl, -O-C _1-3 Alkyl, -O-C _3-6 Cycloalkyl, C _3-6 Cycloalkyl and 3-8 membered heterocyclyl, each of said alkyl, cycloalkyl and heterocyclyl being optionally substituted with one or more substituents selected from hydrogen, halogen, hydroxy, cyano, C _1-3 Alkyl and C _1-3 Substituted with a haloalkyl;

preferably, R ² Is selected from C _6-10 Aryl and 5-10 membered heteroaryl, each optionally substituted with one or more R ⁷ Substituted, each R ⁷ Each independently selected from hydrogen, halogen, hydroxy, C _1-3 Alkyl radical, C _2-4 Alkynyl radical、-O-C _1-3 Alkyl and C _3-6 Cycloalkyl, each of which is optionally substituted with one or more of hydrogen and halogen;

preferably, R ² Is selected from C _6-10 Aryl and 5-10 membered heteroaryl, each optionally substituted with one or more substituents independently selected from halogen, hydroxy, C _1-3 Alkyl and C _2-4 And substituent of alkynyl.

7. The compound of any one of claims 1-6, or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, or metabolite thereof, wherein:

each R ³ Each independently selected from hydrogen, halogen, hydroxy, cyano, C _1-4 Alkyl radical, C _1-4 Haloalkyl, C _2-4 Alkenyl radical, C _2-4 Alkynyl, -O-C _1-4 Alkyl, -O-C _3-6 Cycloalkyl, -O-C _2-4 Alkenyl, -O-C _2-4 Alkynyl, C _3-6 Cycloalkyl and 3-8 membered heterocyclyl; or two R of the same carbon atom ³ And the carbon atoms to which they are both attached form a carbonyl group;

preferably, each R ³ Each independently selected from hydrogen, halogen, hydroxy, C _1-4 Alkyl radical, C _1-4 Haloalkyl, -O-C _1-4 Alkyl, -O-C _3-6 Cycloalkyl radical, C _3-6 Cycloalkyl and 3-8 membered heterocyclyl;

preferably, each R ³ Each independently is halogen;

preferably, each R ³ Each independently selected from F and Cl.

8. The compound of any one of claims 1-7, or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, or metabolite thereof, wherein:

preferably, R ¹¹ Selected from hydrogen, C _1-6 Alkyl and 3-12 membered heterocyclyl;

preferably, R ¹¹ Selected from the group consisting of hydrogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, pentyl and morpholinyl.

9. A compound or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, or metabolite thereof, wherein the compound has the structure of formula (II-a), formula (II-B), formula (II-C), formula (II-D), or formula (II-E):

wherein m is 0, 1 or 2, each group L ¹ 、R ¹ 、R ² 、R ³ 、R ⁴ 、R ⁸ N, o, p, q, r and s are as defined in any one of claims 1 to 8.

10. A compound, or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, or metabolite thereof, said compound having the structure:

11. a pharmaceutical composition comprising a prophylactically or therapeutically effective amount of a compound according to any one of claims 1 to 10, or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, or metabolite thereof, and one or more pharmaceutically acceptable carriers.

12. Use of a compound of any one of claims 1-10, or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound or metabolite, or a pharmaceutical composition of claim 11, for the manufacture of a medicament for the prevention and/or treatment of KRAS G12D mediated related diseases;

preferably, the KRAS G12D-mediated related disease is a tumor.

13. A process for preparing the compound of formula (II-a) of claim 9, comprising the steps of:

wherein L is ¹ 、R ¹ 、R ² 、R ³ 、R ⁸ M, n, o, p, q, r and s are as defined in any one of claims 1 to 8;

LG ¹ 、LG ² 、LG ³ and X represents a leaving group such as a halogen atom, trifluoromethanesulfonate, methylthio, methylsulfinyl or methylsulfonyl group, etc.;

PG ¹ 、PG ² and PG ³ Represents a protecting group such as benzyloxycarbonyl (Cbz), tert-butoxycarbonyl (Boc), methyl or benzyl;

(1) Reacting compound II-A1-1 with compound II-A1-2 to give compound II-A1-3;

(6) Reacting compound II-A-9 to give compound II-A-10;

(9) Subjecting compound II-A-13 to a deprotection reaction to give compound II-A; or

The method comprises the following steps:

PG ¹ and PG ³ Represents a protecting group, bagIncluding but not limited to benzyloxycarbonyl (Cbz), tert-butoxycarbonyl (Boc), benzyl, and the like;

(2) Subjecting compound IIa-A-2 to a coupling reaction with compound II-A-4 to obtain compound IIa-A-3;

(5) Subjecting the compound II-A-13 to a deprotection reaction to obtain the compound II-A.

14. A process for preparing the compound of claim 9 of formula (II-C), comprising the steps of:

(1) Reacting compound II-C1-1 with compound II-C-2 to give compound II-C-3;

(2) Reacting compound II-C-3 with compound II-A-4 to obtain compound II-C-4;

(4) Reacting compound II-C-5 to obtain compound II-C-6;

(5) Carrying out coupling reaction on the compound II-C-6 and the compound II-C-7 to obtain a compound II-C-8;

(7) Carrying out coupling reaction on the compound II-C-9 and the compound II-C-10 to obtain a compound II-C-11;

(8) Subjecting the compound II-C-11 to a deprotection reaction to obtain a compound II-C.