A compound targeting pan-kras protein degradation agent and application thereof

Abstract

A compound targeting KRAS protein degradation agent and application thereof. The compound is a compound with a structure of F-L-M, or a tautomer, a racemate, an enantiomer, a diastereomer, or a mixture thereof, or a pharmaceutically acceptable salt thereof. F is a KRAS protein binding fragment, L is a connecting unit connecting F and M, and M is a VHL binding fragment. The compound can be used for treating diseases caused by KRAS mutation or amplification.

Description

A compound targeting pan-KRAS protein degrader and its application Technical Field

[0001] The present invention belongs to the field of medical technology, and specifically relates to a compound that is a targeted KRAS protein degrader and applications thereof. Background Art

[0002] PROTAC is a heterobifunctional molecule with a ligand that binds to the target protein at one end and a ligand for the E3 ligase at the other end, connected in the middle by a suitable chain. PROTAC degradation of target proteins is achieved through the ubiquitin proteasome system (UPS). The general process is as follows: PROTAC molecules bind to the target protein (POI) and the E3 ligase to form a ternary complex, marking the target protein with ubiquitin molecules. The ubiquitinated protein is recognized and degraded by the proteasome 26S in the cell.

[0003] Compared with traditional small molecules and antibodies, which inhibit the function of target proteins and play a role in treating diseases through an "occupancy-driven" mode of action, this mode of action requires a high concentration of inhibitors or monoclonal antibodies to occupy the active site of the target and block the transduction of downstream signaling pathways. PROTAC is "event-driven" and does not affect the function of the protein, but rather mediates the degradation of pathogenic target proteins. As long as PROTAC mediates the formation of a ternary complex and tags the target protein with ubiquitination, it can theoretically be recycled and reused, so the catalytic amount can play a role. It has the characteristics of a wider range of action, higher activity, and the ability to target "undruggable" targets; improve selectivity, activity and safety; and overcome drug resistance.

[0004] The activity of KRAS switches depending on the binding transition between GDP and GTP. It is inactive when bound to GDP and activated when bound to GTP. In mutated KRAS proteins, GAP access to GTP is restricted, preventing hydrolysis and forming a persistently activated GTP-bound state. Because mutant KRAS has a high affinity for guanosine triphosphate (GTP), and has difficult-to-target factors such as a small catalytic site and a smooth protein surface, the development of small molecule inhibitors has been challenging, making KRAS "undruggable." On the one hand, KRAS binds to GDP and GTP with an affinity of picomolar concentrations, which seriously hinders the development of nucleotide competitive inhibitors; on the other hand, the KRAS protein lacks an ideal small molecule binding pocket, making it difficult to design high-affinity allosteric inhibitors.

[0005] Cancers caused by RAS mutations account for 25%-30% of all human cancers. KRAS mutations account for 86% of the three RAS mutations. Among all KRAS mutations, there are G12D (35%), G12V (29%), G12C (21%), G13D (6.7%), G12R (4.3%), G12A (4.1%) and other mutations. If these different KRAS mutations can be targeted simultaneously, it will have huge commercial value while greatly meeting clinical needs.

[0006] However, current KRAS inhibitors face the dilemma of limited efficacy and high drug resistance in clinical practice. If the anti-resistance and high efficacy of PROTAC can be combined to develop a new pan-KRAS PROTAC degrader drug for the treatment of diseases caused by KRAS mutation or amplification, it will have great social significance and commercial value.

[0007] Summary of the Invention

[0008] The present invention provides a novel pan-KRAS PROTAC degrader that can be used to treat diseases caused by KRAS mutation or amplification.

[0009] In one aspect, the present invention provides a compound having the structure of FLM, or a tautomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, wherein:

[0010] Said F is a KRAS protein binding fragment represented by general formula (I);

[0011] The K is selected from a 6-12 membered heterocyclic group containing N; the K may be optionally further replaced by one or more R a replaced by;

[0012] The R 14 selected from halogen;

[0013] The R6 is selected from H, hydroxyl, amino, cyano, halogen, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Haloalkoxy or C 1-6 Alkoxy; preferably H;

[0014] Alternatively, the R6 and K and the atoms to which they are connected together form a 3-14 membered heterocyclic group, and the 3-14 membered heterocyclic group may be further optionally replaced by one or more R a replaced by;

[0015] The L is a connecting unit connecting F and M; the L is selected from

[0016] in,

[0017] The G ring and the D ring are each independently selected from C 6-14 Aryl, 5-14 membered heteroaryl, C 3-14 Cycloalkyl or 3-14 membered heterocyclic group, the C 6-14 Aryl, 5-14 membered heteroaryl, C 3-14 The cycloalkyl or 3-14 membered heterocyclic group is optionally further substituted by one or more R a replace;

[0018] The A rings are each independently selected from 3-7 membered N-containing heterocyclic groups; the 3-7 membered N-containing heterocyclic groups are optionally further substituted by one or more R a Preferably, the A ring is independently selected from a 3-6 membered N-containing heterocyclic group; the 3-6 membered N-containing heterocyclic group is optionally further substituted by one or more R a replace;

[0019] The E's are each independently absent or selected from -O-, -NH-, or -NCH3-;

[0020] Said R7, R8 are each independently selected from H, halogen, hydroxyl, amino, oxo, cyano, halogen, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Haloalkoxy or C 1-6 alkoxy;

[0021] Alternatively, R7, R8 and the atoms to which they are connected together form C 3-14 Cycloalkyl or 3-14 membered heterocyclic group; said C 3-14 The cycloalkyl or 3-14 membered heterocyclic group is optionally further substituted with one or more R a replace;

[0022] Said n1, n2, n3, n4 are each independently selected from an integer of 0-10, preferably 0, 1, 2, 3 or 4;

[0023] The M is selected from the VHL binding fragment represented by general formula (II);

[0024] wherein X1 is selected from C, CH or N;

[0025] Said X2 is selected from C, CH or N;

[0026] Said X3 is selected from N, NH, O or S;

[0027] Said R1 is selected from H, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C1-6 Alkoxy, C 1-6 Haloalkoxy, halogen or cyano, preferably methyl, ethyl, F or cyano;

[0028] Said R2 is selected from H, C 1-6 Alkyl or C 1-6 Haloalkyl, preferably H or methyl;

[0029] Said R3 is selected from H, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Hydroxyalkyl, -(CH2) n N(R9)2、-(CH2) n -CO-OR9 or -(CH2) n -CO-N(R9)2, wherein R9 are independently selected from H, C 1-6 Alkyl, C 1-6 Haloalkyl or C 1-6 Alkoxy; preferably, the -(CH2) n -CO-N(R9)2 is -(CH2) n -CO-NH-(CH2) n -CH3;

[0030] Said n is selected from 0, 1, 2 or 3;

[0031] The R4 is selected from or -NH-;

[0032] Said R5 is selected from H, C 1-6 Alkyl or C 1-6 alkyl halide;

[0033] The R a are independently selected from H, hydroxy, amino, oxo, cyano, halogen, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Alkoxy, -C 0-3 Alkylene-OR b 、-OC(=O)C 1-6 Alkyl, -C 0-3 Alkylene-SR b 、-C 0-3 Alkylene-N(R b )2. -C 0-3 Alkylene-S(=O)R b 、-C 0-3 Alkylene-S(=O)2R b 、-C 0-3 Alkylene-SR b 、-C 0-3Alkylene-S(R b )5. -C 0-3 Alkylene-C(=O)R b 、-C 0-3 Alkylene-C(=O)OR b 、-C 0-3 Alkylene-C(=O)N(R b )2、C 2-6 Alkenyl, C 2-6 Alkynyl, -C 0-3 Alkylene-C 3-14 Cycloalkyl, -C 0-3 Alkylene-(3-14 membered heterocyclic group), -C 0-3 Alkylene-C 6-18 Aryl or -C 0-3 Alkylene-(5-18 membered heteroaryl), the hydroxyl, amino, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Alkoxy, C 0-3 Alkylene, C 2-6 Alkenyl, C 2-6 Alkynyl, -C 0-3 Alkylene-C 3-14 Cycloalkyl, -C 0-3 Alkylene-(3-14 membered heterocyclic group), -C 0-3 Alkylene-C 6-18 Aryl or -C 0-3 Alkylene-(5-18 membered heteroaryl) is optionally further substituted with one or more R b replace;

[0034] Each R b are independently H, halogen, hydroxy, amino, cyano, C 1-6 Alkyl, C 3-6 Cycloalkyl, C 1-6 Halogenated alkyl, C 3-14 Cycloalkyl, 3-14 membered heterocyclic group, C 6-18 Aryl or 5-18 membered heteroaryl, the hydroxyl, amino, C 1-6 Alkyl, C 3-6 Cycloalkyl, C 1-6 Halogenated alkyl, C 3-14 Cycloalkyl, 3-14 membered heterocyclic group, C 6-18 Aryl or 5-18 membered heteroaryl is optionally further substituted with one or more halogen, C 1-6 Alkyl or C 1-6 substituted by a haloalkyl group; or two R b Together with its common atoms, it forms C 3-14 Cycloalkyl, 3-14 membered heterocyclic group, C6-18 Aryl or 5-18 membered heteroaryl, the C 3-14 Cycloalkyl, 3-14 membered heterocyclic group, C 6-18 Aryl or 5-18 membered heteroaryl is optionally further substituted with one or more halogen, amino, hydroxyl, cyano, C 1-6 Alkyl, C 3-6 Cycloalkyl or C 1-6 Haloalkyl substitution.

[0035] In some embodiments of the present invention, the K is selected from

[0036] In some embodiments of the present invention, the F is selected from

[0037] In some embodiments of the present invention, the R4 is

[0038] In some embodiments of the present invention, the R4 is

[0039] In some embodiments of the present invention, said R4 is -NH-.

[0040] In some embodiments of the present invention, the G ring or D ring in L is independently selected from C 6-14 Aryl, 5-14 membered heteroaryl, C 3-14 Cycloalkyl or 3-14 membered heterocyclic group, the C 6-14 Aryl, 5-14 membered heteroaryl, C 3-14 The cycloalkyl or 3-14 membered heterocyclic group is optionally further substituted with one or more halogen or C 1-3 Alkyl substitution;

[0041] The C 6-14 The aryl group is preferably

[0042] The 5-14 membered heteroaryl group is preferably selected from

[0043] The 3-14 membered heterocyclic group is preferably selected from

[0044] The C 3-14 The cycloalkyl group is preferably selected from

[0045] In some embodiments of the present invention, the D ring is independently selected from a 5-6 membered nitrogen-containing heteroaryl group, and the 5-6 membered nitrogen-containing heteroaryl group is optionally further substituted by one or more R areplace;

[0046] The G rings are each independently selected from C 6-14 Aryl, 5-14 membered heteroaryl, C 3-14 Cycloalkyl or 3-14 membered heterocyclic group, the C 6-14 Aryl, 5-14 membered heteroaryl, C 3-14 The cycloalkyl or 3-14 membered heterocyclic group is optionally further substituted by one or more R a replace;

[0047] The A rings are each independently selected from 3-7 membered N-containing heterocyclic groups; the 3-7 membered N-containing heterocyclic groups are optionally further substituted by one or more R a replace;

[0048] The E's are each independently absent or selected from -O-, -NH-, or -NCH3-;

[0049] Said R7, R8 are each independently selected from H, halogen, hydroxyl, amino, oxo, cyano, halogen, C 1-6 Alkyl, C 1-6 Haloalkyl or C 1-6 alkoxy;

[0050] Alternatively, the R7, R8 and the atoms to which they are connected together form a C 3-14 Cycloalkyl or 3-14 membered heterocyclic group; said C 3- 14 The cycloalkyl or 3-14 membered heterocyclic group is optionally further substituted with one or more R a replace;

[0051] Said n1, n2, n3 and n4 are each independently selected from integers of 0-10, preferably 0, 1, 2, 3 or 4.

[0052] In some embodiments of the present invention, the A rings are each independently selected from 3-7 membered N-containing heterocyclic groups; the 3-7 membered N-containing heterocyclic groups are optionally further substituted with one or more halogens or C 1-3 Alkyl substitution.

[0053] Furthermore, the 3-7 membered N-containing heterocyclic group is

[0054] In some embodiments of the present invention, E is -O-.

[0055] In some embodiments of the present invention, E is -NH-.

[0056] In some embodiments of the present invention, E is -NCH3-.

[0057] In some embodiments of the present invention, the D ring is independently selected from a 5-6 membered nitrogen-containing heteroaryl group, and the 5-6 membered heteroaryl group is selected from The 5-6 membered nitrogen-containing heteroaryl group is optionally further substituted with one or more R a Substitution; said R a Selected from H, hydroxy, amino, oxo, cyano, halogen, C 1-6 Alkyl, C 1-6 Haloalkyl or C 1-6 Alkoxy.

[0058] In some embodiments of the present invention, the G ring is independently selected from C 6-14 Aryl, 5-14 membered heteroaryl, C 3-14 Cycloalkyl or 3-14 membered heterocyclic group, the C 6-14 Aryl, 5-14 membered heteroaryl, C 3-14 The cycloalkyl or 3-14 membered heterocyclic group is optionally further substituted by one or more R a replace;

[0059] The C 6-14 The aryl group is preferably

[0060] The 5-14 membered heteroaryl group is preferably selected from

[0061] The 3-14 membered heterocyclic group is preferably selected from

[0062] The C 3-14 The cycloalkyl group is preferably selected from

[0063] The R a Selected from H, hydroxy, amino, oxo, cyano, halogen, C 1-6 Alkyl, C 1-6 Haloalkyl or C 1-6 Alkoxy.

[0064] In some embodiments of the present invention, the G rings are each independently selected from 3-14 membered heterocyclic groups, wherein the 3-14 membered heterocyclic groups are optionally further substituted by one or more R a Substituted; the 3-14 membered heterocyclic group is preferably selected from

[0065] The R a Selected from H, hydroxy, amino, oxo, cyano, halogen, C 1-6 Alkyl, C 1-6Haloalkyl or C 1-6 Alkoxy.

[0066] In some embodiments of the present invention, the L is selected from

[0067] In some embodiments of the present invention, the L is selected from

[0068] In some embodiments of the present invention, R3 is selected from H, methyl, hydroxymethyl, -CH2-CO-O-CH3, -CH2-N(CH3)2 or -CH2-CO-NH-CH3.

[0069] In some embodiments of the present invention, the M is selected from

[0070] In some embodiments of the present invention, the compound of FLM structure is selected from the following compounds:

[0071] On the other hand, the present invention provides a pharmaceutical composition comprising any compound represented by the general formula having an FLM structure of the present invention, its tautomers, racemates, enantiomers, diastereomers, or mixtures thereof, or pharmaceutically acceptable salts thereof, and one or more pharmaceutically acceptable carriers, diluents or excipients.

[0072] The present invention further provides the use of any of the above-mentioned compounds represented by the general formula having an FLM structure, its tautomers, racemates, enantiomers, diastereomers, or mixtures thereof, or pharmaceutically acceptable salts thereof, or pharmaceutical compositions containing the same, in the preparation of a medicament for regulating the ubiquitination and degradation of KRAS protein in a subject.

[0073] The present invention further provides the use of any of the above-mentioned compounds having the general structural formula FLM or its tautomers, racemates, enantiomers, diastereomers, or mixtures thereof or pharmaceutically acceptable salts, or pharmaceutical compositions comprising the same, in the preparation of medicaments for treating and / or preventing KRAS-mediated or dependent diseases, wherein the KRAS-mediated disease is preferably selected from tumors.

[0074] In certain embodiments, the disease is selected from breast cancer, multiple myeloma, bladder cancer, endometrial cancer, gastric cancer, cervical cancer, rhabdomyosarcoma, non-small cell lung cancer, small cell lung cancer, pleomorphic lung cancer, ovarian cancer, esophageal cancer, melanoma, colorectal cancer, hepatocellular carcinoma, head and neck cancer, hepatobiliary carcinoma, myelodysplastic syndrome, malignant glioma, prostate cancer, thyroid cancer, Schwann cell tumor, squamous cell carcinoma of the lung, lichenoid keratosis, synovial sarcoma, skin cancer, pancreatic cancer, testicular cancer, or liposarcoma.

[0075] Unless otherwise indicated, general chemical terms used in the structural formulae have their usual meanings.

[0076] For example, the term "halogen," as used herein, refers to fluorine, chlorine, bromine, or iodine, unless otherwise indicated.

[0077] Unless otherwise specified, when the linking group involved in the present invention does not specify its connection direction, its connection direction is arbitrary. For example, when the L of the linking group in the FLM compound is selected from When L is connected from left to right, F and M can be connected to form You can also connect F and M from right to left to form

[0078] Preferably, in the connection order of L of the present invention, the D ring is close to M (VHL end) and the G ring is close to F (KRAS end).

[0079] In the present invention, unless otherwise specified, "alkyl" includes a linear or branched monovalent saturated hydrocarbon group. For example, alkyl includes methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, 3-(2-methyl)butyl, 2-pentyl, 2-methylbutyl, neopentyl, n-hexyl, 2-hexyl, 2-methylpentyl, etc. Similarly, "C 1-6 The "1-6" in "alkyl" refers to a group containing 1, 2, 3, 4, 5 or 6 carbon atoms in a straight chain or branched form.

[0080] The term "alkoxy" refers to the oxygen ether form of the aforementioned straight-chain or branched alkyl groups, ie, -O-alkyl.

[0081] The term "alkylene" refers to a divalent alkyl linking group. Alkylene formally refers to an alkane with two C—H bonds replaced as the point of attachment of the alkylene to the rest of the compound. Similarly, C 1-3 The "C" in the alkylene 1-3 ” refers to an alkylene group containing 1, 2 or 3 carbon atoms, including but not limited to methylene, 1,2-ethylene, 1,3-propylene or 1,2-isopropylene.

[0082] The term "haloalkyl" refers to an alkyl group in which one or more H groups have been replaced by a halogen atom.

[0083] The term "oxo" or "oxo group" refers to an oxygen atom in the form of a divalent substituent, which forms a carbonyl group when attached to C, and forms a sulfoxide group or a sulfone group or an N-oxide group when attached to a heteroatom.

[0084] In the present invention, unless otherwise specified, the term "aromatic ring", "aromatic ring" or "aromatic heterocycle" refers to a polyunsaturated carbon ring or heterocycle with aromatic characteristics (having (4n+2) delocalized π electrons, where n is an integer).

[0085] The term "aryl", in the present invention, unless otherwise specified, refers to an unsubstituted or substituted monocyclic or condensed ring aromatic group containing carbon ring atoms. 6-18 Aryl, more preferably C 6-10 A monocyclic or bicyclic aromatic ring group. Phenyl or naphthyl is preferred; naphthyl is most preferred. The aryl ring may be fused to a heteroaryl, heterocyclic, or cycloalkyl group, wherein the ring connecting to the parent structure is an aryl ring. Non-limiting examples include, but are not limited to, benzocyclopentyl.

[0086] The term "heterocyclyl" refers to a ring system having at least one cyclized alkyl or cyclized alkenyl group containing a heteroatom, wherein the heteroatom is selected from N, O and / or S. The heterocyclyl may include a monocyclic or polycyclic ring (for example, having 2, 3 or 4 fused rings, spirocyclic rings, bridged rings, etc.). The heterocyclyl may be connected to the other parts of the compound via a ring-forming carbon atom or a ring-forming heteroatom. Preferably, it is a 3-14-membered heterocyclyl, more preferably a 3-8-membered heterocyclyl, a 3-7-membered heterocyclyl, or a 3-6-membered heterocyclyl; wherein the "3-14 members" in the 3-14-membered heterocyclyl refers to a heterocyclyl composed of 3-14 C, N, O or S ring atoms; wherein the nitrogen or sulfur heteroatom can be selectively oxidized, and the nitrogen heteroatom can be selectively quaternized. Examples of these heterocyclyls include, but are not limited to Pyrrolidinyl, piperidinyl, piperazinyl, oxopiperazinyl, oxopiperidinyl, tetrahydrofuranyl, dioxolanyl, tetrahydroimidazolyl, tetrahydrothiazolyl, tetrahydrooxazolyl, tetrahydropyranyl, morpholinyl, thiomorpholinyl, thiomorpholinyl sulfoxide, thiomorpholinyl sulfone and tetrahydrooxadiazolyl. The heterocyclic group can be fused to an aryl, heteroaryl or cycloalkyl ring, wherein the ring connected to the parent structure is a heterocyclic group.

[0087] The term "heteroaryl" as used herein, unless otherwise specified, refers to a monocyclic or polycyclic (e.g., 2, 3, or 4 fused, spiro, or bridged) aromatic heterocycle having at least one heteroatom, wherein the heteroatom is selected from N, O, and / or S, and wherein the nitrogen or sulfur heteroatom may be selectively oxidized, and the nitrogen heteroatom may be selectively quaternized. Preferably, the heteroaryl is a 5-18-membered heteroaryl, more preferably a 5-10-membered heteroaryl or a 5-6-membered heteroaryl; wherein the "5-18 members" in the 5-18-membered heteroaryl refers to a heteroaryl group consisting of 5-18 carbon, nitrogen, oxygen, or sulfur atoms in the ring. More preferred is a 5-10-membered heteroaryl; even more preferred is a 5-6-membered heteroaryl. Examples of heteroaryl groups include, but are not limited to, thienyl, furanyl, imidazolyl, isoxazolyl, oxazolyl, pyrazolyl, pyrrolyl, thiazolyl, thiadiazolyl, triazolyl, pyridyl, pyridazinyl, indolyl, azaindolyl, indazolyl, benzimidazolyl, benzofuranyl, benzothienyl, benzisoxazolyl, benzothiazolyl, benzothiadiazolyl, benzotriazolyladenine, quinolyl, or isoquinolyl. The heteroaryl group may be fused to an aryl, heterocyclyl, or cycloalkyl ring, wherein the ring attached to the parent structure is the heteroaryl ring.

[0088] The term "cycloalkyl" refers to a ring system having at least one cyclized alkyl group. 3-14 Cycloalkyl, more preferably C 3-8 Cycloalkyl, C 3-6 Cycloalkyl, C 5-7 Cycloalkyl; wherein "C 3-14 " means that the cycloalkyl group may have 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 or 14 ring atoms. The cycloalkyl group may include monocyclic and polycyclic rings (e.g., having 2, 3 or 4 fused rings, spiro rings, bridged rings, etc.). In some embodiments, the cycloalkyl group includes but is not limited to cyclopropyl, cyclobutyl, cyclopentyl, The cycloalkyl group can also be fused to an aryl, heterocyclyl or heteroaryl ring, wherein the ring connected to the parent structure is a cycloalkyl group.

[0089] The term "substituted" means that one or more hydrogen atoms in a group are replaced by the same or different substituents. Typical substituents include but are not limited to halogen (F, Cl, Br or I), C 1-8 Alkyl, C 3-12 Cycloalkyl, -OR 1 、-SR 1 , =O, =S, -C(O)R 1 、-C(S)R 1 、=NR 1 、-C(O)OR 1 、-C(S)OR 1 、-NR 1 R2 、-C(O)NR 1 R 2 , cyano, nitro, -S(O)2R 1 、-OS(O2)OR 1 、-OS(O)2R 1 、-OP(O)(OR 1 )(OR 2 ); where R 1 and R 2 Independently selected from -H, C 1-6 Alkyl, C 1-6 Haloalkyl or C 3-6 In some embodiments, the substituents are independently selected from the group consisting of -F, -Cl, -Br, -I, -OH, trifluoromethoxy, ethoxy, propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, -SCH3, -SC2H5, formaldehyde, -COCH3, cyano, nitro, -CF3, amino, dimethylamino, sulfonyl, and acetyl.

[0090] When the number of a linking group is 0, such as -(CH2)0-, it means that the linking group is a bond.

[0091] The term "pharmaceutically acceptable salts" refers to salts prepared from pharmaceutically acceptable non-toxic bases or acids.

[0092] When the compound provided by the present invention is an acid, its corresponding salt can be easily prepared from pharmaceutically acceptable nontoxic bases, including inorganic bases and organic bases. Salts derived from inorganic bases include salts of aluminum, ammonium, calcium, copper (high and low valence), ferric iron, ferrous iron, lithium, magnesium, manganese (high and low valence), potassium, sodium, zinc and the like. Particularly preferred are salts of ammonium, calcium, magnesium, potassium and sodium. Nontoxic organic bases that can be derived into pharmaceutically acceptable salts include primary amines, secondary amines and tertiary amines, as well as cyclic amines and substituted amines, such as naturally occurring and synthetic substituted amines. Other pharmaceutically acceptable non-toxic organic bases capable of forming salts include ion exchange resins and arginine, betaine, caffeine, choline, N',N'-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, reduced glucosamine, glucosamine, histidine, isopropylamine, lysine, methylglucamine, morpholine, piperazine, piperidine, polyamine resins, procaine, chloroprocaine, purine, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine, and the like.

[0093] When compound provided by the invention is a base, it is possible to conveniently prepare its corresponding salt from pharmaceutically acceptable nontoxic acid, including inorganic and organic acids. Such acid includes, as, acetic acid, benzenesulfonic acid, benzoic acid, camphorsulfonic acid, citric acid, ethanesulfonic acid, formic acid, fumaric acid, gluconic acid, glutamic acid, hydrobromic acid, hydrochloric acid, isethionic acid, lactic acid, maleic acid, malic acid, mandelic acid, methanesulfonic acid, mucic acid, nitric acid, tamoxifen, pantothenic acid, phosphoric acid, succinic acid, sulfuric acid, oxalic acid, propionic acid, glycolic acid, hydroiodic acid, perchloric acid, cyclamic acid, salicylic acid, 2-naphthalenesulfonic acid, saccharinic acid, trifluoroacetic acid, tartaric acid and p-toluenesulfonic acid etc. Preferably, citric acid, hydrobromic acid, formic acid, hydrochloric acid, maleic acid, phosphoric acid, sulfuric acid and tartaric acid. More preferably, formic acid and hydrochloric acid.

[0094] Prodrugs of the compounds of the present invention are included within the scope of protection of the present invention. Generally, such prodrugs are functional derivatives that are readily converted into the desired compound in vivo. For example, any pharmaceutically acceptable salt, ester, ester salt, or other derivative of the compounds of the present invention, which, upon administration to a recipient, can directly or indirectly provide the compounds of the present invention or their pharmaceutically active metabolites or residues.

[0095] The compounds of the present invention may contain one or more asymmetric centers and may thus produce diastereomers and optical isomers. The present invention includes all possible diastereomers and racemic mixtures thereof, their substantially pure resolved enantiomers, all possible geometric isomers and pharmaceutically acceptable salts thereof.

[0096] When the compound represented by formula (I) exists in tautomers, unless otherwise stated, the present invention includes any possible tautomers and pharmaceutically acceptable salts thereof, and mixtures thereof.

[0097] The term "pharmaceutical composition" refers to a mixture of one or more compounds of the present application or pharmaceutically acceptable salts thereof and pharmaceutically acceptable excipients. The purpose of a pharmaceutical composition is to facilitate administration of the compounds of the present application to an organism.

[0098] In the present invention, "a", "an", "the", "at least one" and "one or more" are used interchangeably. Thus, for example, a mixture comprising "a" pharmaceutically acceptable excipient composition can be interpreted as indicating that the pharmaceutical composition includes "one or more" pharmaceutically acceptable excipients.

[0099] The term "pharmaceutically acceptable excipient" refers to an excipient that is non-irritating to organisms and does not impair the biological activity and properties of the active compound. Suitable excipients are well known to those skilled in the art and include, for example, carbohydrates, waxes, water-soluble and / or water-swellable polymers, hydrophilic or hydrophobic materials, gelatin, oils, solvents, water, and the like.

[0100] The pharmaceutical compositions of the present invention can be prepared by combining the compounds of the present application with suitable pharmaceutically acceptable excipients, and can be formulated into solid, semi-solid, liquid or gaseous preparations, such as tablets, pills, capsules, powders, granules, ointments, emulsions, suspensions, suppositories, injections, inhalants, gels, microspheres and aerosols.

[0101] Typical routes of administration of the compounds of the present invention, or pharmaceutically acceptable salts thereof, or pharmaceutical compositions thereof include, but are not limited to, oral, rectal, topical, inhalation, parenteral, sublingual, vaginal, intranasal, intraocular, intraperitoneal, intramuscular, subcutaneous, and intravenous administration.

[0102] The term "treat" generally refers to obtaining a desired pharmacological and / or physiological effect. This effect can be therapeutic in terms of partial or complete stabilization or cure of a disease and / or side effects caused by the disease. As used herein, "treat" encompasses any treatment of a patient's disease, including: (a) suppressing the symptoms of the disease, i.e., arresting its progression; or (b) relieving the symptoms of the disease, i.e., causing regression of the disease or its symptoms.

[0103] The term "effective amount" means an amount of a compound of the present invention that (i) treats or prevents a specific disease, condition, or disorder, (ii) alleviates, ameliorates, or eliminates one or more symptoms of a specific disease, condition, or disorder, or (iii) prevents or delays the onset of one or more symptoms of a specific disease, condition, or disorder described herein. The amount of a compound of the present invention that constitutes a "therapeutically effective amount" varies depending on the compound, the disease state and its severity, the mode of administration, and the age of the mammal to be treated, but can be routinely determined by those skilled in the art based on their own knowledge and the present disclosure.

[0104] The room temperature mentioned in the present invention refers to the temperature of the indoor environment, which is generally 18-25°C.

[0105] The compounds of the present invention have excellent performance in biological activities such as KRAS degradation ability, cell proliferation inhibition activity, and in vivo pharmacokinetic properties, and have high drugability. In addition, through research, it was found that the connecting chain of the compound of the present invention has a great influence on the KRAS degradation ability and cell proliferation inhibition activity of the compound. When a 5-6-membered N-containing heteroaryl group (especially a pyrazine ring and a pyridine ring) is added to the connecting chain, close to the VHL ligand side, the compound's ability to degrade KRAS is enhanced, while the compound's cell proliferation inhibition activity is increased, achieving unexpected results. We speculate that the N-containing heteroaromatic ring is in a special spatial position, and it will interact with the VHL protein through hydrogen bonds and other interactions, thereby increasing the binding force between the compound and VHL, thereby increasing the compound's ability to degrade the KRAS protein. In particular, the use of a specific connecting chain The compound of the present invention formed when the pyrazine end is bonded to the VHL ligand has better KRAS degradation ability and cell proliferation inhibition activity. BRIEF DESCRIPTION OF THE DRAWINGS

[0106] Figure 1 shows the Western Blotting results of compound 115 on KRAS protein degradation in different cell lines in biological experiment 2. DETAILED DESCRIPTION

[0107] To make the above content clearer and more specific, the present invention will further illustrate the technical solutions of the present invention with the following examples. The following examples are only used to illustrate the specific embodiments of the present invention so that those skilled in the art can understand the present invention, but are not intended to limit the scope of protection of the present invention. In the specific embodiments of the present invention, technical means or methods not specifically described are conventional technical means or methods in the art.

[0108] Unless otherwise stated, all temperatures herein are in degrees Celsius.

[0109] The following abbreviations are used in the examples:

[0110] DMF: N,N-dimethylformamide;

[0111] NIS: N-iodosuccinimide;

[0112] THF: tetrahydrofuran;

[0113] EA: ethyl acetate;

[0114] PE: petroleum ether;

[0115] DCM: dichloromethane;

[0116] MeOH: methanol;

[0117] DPPA: diphenylphosphoryl azide;

[0118] CDI: N,N'-carbonyldiimidazole;

[0119] DIEA / DIPEA: N,N-diisopropylethylamine;

[0120] POCl3: phosphorus oxychloride;

[0121] DMSO: dimethyl sulfoxide;

[0122] HATU: 2-(7-Azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate;

[0123] TFA: trifluoroacetic acid;

[0124] DABCO: triethylenediamine;

[0125] TBAF: tetrabutylammonium fluoride;

[0126] TEA: triethylamine;

[0127] TMSOI: trimethylsulfoxide iodide;

[0128] ACN / MeCN: acetonitrile;

[0129] Dioxane: 1,4-dioxane;

[0130] PdCl2(PPh3)2: bis(triphenylphosphine)palladium dichloride;

[0131] Cs2CO3: cesium carbonate;

[0132] DMP: Dess-Martin periodinane;

[0133] CuI: cuprous iodide;

[0134] K3PO4: potassium phosphate;

[0135] ZnCl2: zinc chloride;

[0136] CataCXium A Pd G3: methanesulfonic acid [n-butyldi(1-adamantyl)phosphine] (2-amino-1,1'-biphenyl-2-yl) palladium (II). Synthesis of intermediate M1:

[0137] Step 1: Synthesis of compound M1-1

[0138] At room temperature, 2-chloro-3-fluoro-pyridine-4-carboxylic acid (54.00 g), toluene (390.00 mL), tert-butyl alcohol (390.00 mL), triethylamine (128.27 mL), powdered Molecular sieves (90.00 mL) (pre-activated), under nitrogen protection, reflux for half an hour (internal temperature 87°C). Then cool naturally to room temperature, then add DPPA (99.44 mL), raise the temperature to reflux, and keep the reaction for 5 hours. The reaction mixture is cooled to below 40°C, then diluted with EA500 mL; continue to cool to room temperature, filter with diatomaceous earth to remove the added molecular sieves; rinse the filter residue with EA1500 mL several times and drain; collect the filtrate, wash with water 700 mL and saturated brine 700 mL in turn, separate the liquids; dry the organic phase with anhydrous sodium sulfate; filter, remove the desiccant, concentrate, and purify the concentrate by column chromatography (PE / EA=30:1~20:1), and concentrate the eluent to finally obtain compound M1-1 (68.2 g, yield 89.88%).

[0139] ESI-MS m / z:247.1[M+H] + .

[0140] Step 2: Synthesis of Compound M1-2

[0141] At room temperature, compound M1-1 (65.00 g) was dissolved in CH3CN (82.00 mL), cooled in a water bath, and hydrochloric acid (4M / dioxane) (264 mL) was slowly added. The reaction was stirred at room temperature for about 16 hours, and a white solid precipitated in a suspended state. The reaction mixture was filtered, and the filter cake was rinsed with a small amount of acetonitrile, drained, and the filtrate discarded. The filter cake was collected and added to a mixture of 700 mL of saturated sodium bicarbonate aqueous solution and 700 mL of ethyl acetate, alkalized, extracted, and separated; the aqueous phase was further extracted with 350 mL of ethyl acetate and separated; the ethyl acetate phases were combined, washed with 300 mL of saturated sodium chloride aqueous solution, and separated; the organic phase was dried over anhydrous sodium sulfate, filtered, the desiccant was removed, and concentrated to obtain compound M1-2 (36.3 g, 94.0% yield).

[0142] ESI-MS m / z:147.1[M+H] + .

[0143] Step 3: Synthesis of Compound M1-3

[0144] Compound M1-2 (36.00 g) was dissolved in acetonitrile (180.00 mL) at room temperature, and NIS (66.32 g) and p-toluenesulfonic acid (2.12 g) were added. The mixture was heated to 70°C under nitrogen. The reaction mixture was cooled to 50°C, and 900 mL of water was added. A white solid precipitated and was slurried for half an hour. The mixture was filtered, and the filter cake was rinsed with water and dried. The filter cake was collected and completely dissolved in 1200 mL of ethyl acetate. The mixture was then washed twice with 350 mL of saturated sodium sulfite solution and then with 350 mL of saturated brine. The mixture was separated, and the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain compound M1-3 (63.2 g, 94.43% yield).

[0145] ESI-MS m / z:272.9[M+H] + .

[0146] Step 4: Synthesis of Compound M1-4

[0147] At room temperature, compound M1-3 (57.50 g) was dissolved in DMF (22.00 mL), zinc cyanide (32.22 g), tetrakistriphenylphosphine palladium (12.19 g) and powdered Molecular sieves (20.00 mL) were added, and the mixture was heated to 100°C under a nitrogen atmosphere for approximately 7 hours. The oil bath was removed, and the mixture was allowed to cool naturally to room temperature for post-processing. The reaction mixture was filtered using diatomaceous earth as a filter aid and drained. The filtrate was collected and concentrated at 60-70°C to obtain a pale yellow solid crude product. The filter residue was rinsed with 500 mL of ethyl acetate and drained. The rinse was collected and added to the crude product, and concentrated again until no liquid was distilled out. The concentrated crude solid product was dissolved in 700 mL of ethyl acetate, and then washed three times with 250 mL of saturated sodium chloride each time, and the layers were separated. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a pale yellow solid. 160 mL of a 3 / 1 PE / EA mixture was added, and the mixture was slurried for half an hour, filtered, and drained. The filter cake was collected, water bathed at 45°C, concentrated, and then pumped to constant weight with a high vacuum oil pump to obtain compound M1-4 (36.1 g, 99.7% yield).

[0148] ESI-MS m / z:172.0[M+H] + .

[0149] Step 5: Synthesis of Compound M1-5

[0150] At room temperature, concentrated sulfuric acid (61.37 mL) was added to a 500 mL single-necked flask, cooled to below 10°C in an ice-water bath, and compound M1-4 (39.30 g) was added in batches. After the addition, the mixture was stirred for 10 minutes. In a nitrogen atmosphere, the mixture was kept at 60°C in an oil bath and reacted for about 1 hour. The reaction solution was cooled to room temperature and then carefully added to 1100 mL of ice-water mixture, diluted and quenched, and a small amount of yellow solid precipitated. After stirring for 10 minutes, the mixture was filtered; the filter cake was collected and slurried with 50 mL of saturated sodium bicarbonate aqueous solution for 20 minutes, filtered again, and the two filtrates were collected and combined; then sodium carbonate solid was slowly added to adjust the pH to approximately 7, and a white solid powder precipitated. Stir for half an hour, filter, and drain; rinse the filter cake with 100 mL of water each time and drain, for a total of rinsing twice. The filter cake was collected and placed in a vacuum oven and dried at 55°C to constant weight to obtain compound M1-5 (33.6 g, yield 77.37%).

[0151] ESI-MS m / z:190.0[M+H] + .

[0152] Step 6: Synthesis of Compound M1-6

[0153] At room temperature, tetrahydrofuran (470.00 mL) was added. After nitrogen replacement, sodium hydride (10.00 g) was added under a slight nitrogen flow. The mixture was heated in an oil bath at 40-45°C and stirred for 15 minutes. Compound M1-5 (18.95 g) was then added in portions. After addition, the mixture was mechanically stirred for 20 minutes. CDI (24.31 g) was then carefully added in portions. After addition, the mixture was stirred for 15 minutes, and the mixture was heated in an oil bath and refluxed. The reaction mixture was cooled to below 10°C in an ice-water bath. 500 mL of saturated aqueous ammonium chloride was then added. A light yellow solid precipitated. 1000 mL of water was added. The mixture was then transferred to a 5 L beaker and 3000 mL of water was added. The mixture was stirred for 1 hour, filtered, and drained. The filter cake was collected and dried in a vacuum oven at 50-55°C to constant weight to obtain compound M1-6 (18.3 g, 84.93% yield).

[0154] ESI-MS m / z:216.0[M+H] + .

[0155] Step 7: Synthesis of Compound M1

[0156] Compound M1-6 (18.00 g) and DIEA (36.00 mL) were dissolved in POCl3 (180.00 mL) at room temperature and heated to 100°C under a nitrogen atmosphere for approximately 2.5 hours. The mixture was concentrated under reduced pressure to remove phosphorus oxychloride and rinsed twice with 100 mL of DCM. The residue was dissolved in 400 mL of dichloromethane and then added dropwise to 500 mL of saturated aqueous sodium bicarbonate solution, cooled with ice water. After stirring for 15 minutes, the mixture was separated. The aqueous phase was extracted with 300 mL of dichloromethane and separated. The combined dichloromethane phases were washed with 300 mL of saturated aqueous sodium chloride solution and separated. The mixture was dried over anhydrous sodium sulfate, filtered, and concentrated. The concentrate was purified on a silica gel column (PE / EA = 90 / 10 to 75 / 25) to obtain compound M1 (10.95 g, 51.94% yield).

[0157] ESI-MS m / z:251.9[M+H] + .

[0158] Synthesis of intermediate M2

[0159] Step 1: Synthesis of compound M2-1

[0160] Compound TMSOI (8.5 g) was dissolved in DMSO (20 mL), cooled to 0°C, and NaH (1.6 g) was added. The N2 was replaced three times, and the reaction was stirred at room temperature for 1.0 h. A solution of compound 3-oxopiperidine-1-carboxylic acid benzyl ester (9.0 g) dissolved in DMSO (5 mL) was slowly added dropwise, and the reaction was stirred at room temperature overnight. After the reaction was complete, the mixture was extracted with EA and H2O, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, dried, and purified by column chromatography to obtain compound M2-1 (3.2 g). ESI-MS m / z: 248.3 [M+H] + .

[0161] Step 2: Synthesis of compound M2-2

[0162] Compound M2-1 (3.2 g), sodium cyanide (1.3 g), anhydrous ethanol (10 mL), and water (10 mL) were replaced with N2 three times and stirred at room temperature for 5 h. After the reaction was complete, the mixture was extracted with EA and H2O, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, dried, and purified by column chromatography (PE:EA 43%) to obtain compound M2-2 (3.0 g). ESI-MS m / z: 275.3 [M+H] + .

[0163] Step 3: Synthesis of Compound M2-3

[0164] Compound M2-2 (1.5 g), palladium hydroxide (0.2 g), and anhydrous ethanol (10 mL) were replaced with hydrogen five times and stirred at room temperature for 1.5 h. After the reaction was complete, the mixture was filtered and the solvent was dried to obtain compound M2-3 (0.9 g), which was directly used for the next step. ESI-MS m / z: 141.3 [M+H] + .

[0165] Step 4: Synthesis of Compound M2

[0166] Compound M1 (1.6 g) and DIPEA (3.5 mL) were dissolved in DCM (15 mL), cooled to below -40°C, and stirred for 10 minutes. Compound M2-3 (0.9 g) was then added and stirred for 1 hour at the same temperature. After the reaction was complete, the mixture was quenched with saturated ammonium chloride solution, extracted with EA and H2O, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, dried, and purified by column chromatography (EA:DCM 12%) to obtain compound M2 (1.2 g). ESI-MS m / z: 356.3 [M+H] + .

[0167] Synthesis of intermediate M3:

[0168] Step 1: Synthesis of Compound M3-1

[0169] ((2-Fluoro-6-(methoxymethoxy)-8-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalen-1-yl)ethynyl)triisopropylsilane (1.0 g) was added to a 50 mL single-necked flask, followed by 6 mL of DMF and cesium fluoride (5.9 g). The mixture was stirred at room temperature for 1 h. TLC confirmed the complete reaction of the starting material. The reaction solution was slowly added dropwise to water (20 mL), resulting in the precipitation of a white solid. The filter cake was collected by filtration and dried to obtain 0.65 g of a white solid, compound M3-1.

[0170] Step 2: Synthesis of Compound M3

[0171] Compound M3-1 (0.65 g) was added to a 50 mL single-necked vial, along with 6 mL of MeOH and Pd / C (600 mg, 10% purity). The mixture was reacted at room temperature under a hydrogen atmosphere for 20 min. After the reaction was complete, the filtrate was filtered, collected, and concentrated. The concentrate was purified by column chromatography to afford compound M3 (496 mg, 75.5% yield).

[0172] 1 H NMR (500MHz, Methanol-d4) δ7.56(dd,J=8.9,5.8Hz,1H),7.39(d,J=2.7Hz,1H),7.36(d,J=2.7Hz,1H),7.20( t,J=9.2Hz,1H),5.27(s,2H),3.50(s,3H),3.12(qd,J=7.5,2.5Hz,2H),1.44(s,12H),1.26(t,J=7.5Hz,3H).

[0173] Synthesis of intermediate M4:

[0174] Compound M3 (1.0 g) was added to a 50 mL single-necked bottle, followed by 10 mL of hydrochloric acid (4 M / Dioxane). After reacting at room temperature for half an hour, the reaction mixture was neutralized by adding saturated sodium bicarbonate solution and extracted twice with EA. The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated. The concentrate was purified by column chromatography (DCM:ammonia methanol = 10:1) to obtain compound M4 (0.82 g). 1 HNMR (500MHz, CDCl3) δ7.46 (dd, J=8.9, 5.8Hz, 1H), 7.27 (d, J=2.7Hz, 1H), 7.17 (dd, J=12. 5,5.9Hz,1H),7.12(d,J=2.7Hz,1H),3.17–3.07(m,2H),1.45(s,12H),1.29–1.25(m,3H).

[0175] Synthesis of intermediate M5:

[0176] Step 1: Synthesis of compound M5-1

[0177] (S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethanamine (12.7 g), Boc-L-hydroxyproline (13.5 g), and HATU (23.2 g) were dissolved in DMF (100 mL). DIEA (38.5 mL) was added and the mixture was reacted at room temperature for 10 minutes. The reaction was monitored for completion by LCMS. EA (100 mL) and saturated brine (100 mL) were added to the reaction solution, and the organic phase was collected, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to obtain 22.0 g of a white solid, compound M5-1.

[0178] ESI-MS m / z:432[M+H] + .

[0179] Step 2: Synthesis of compound M5-2

[0180] Compound M5-1 (22.0 g) was added to DCM (100 mL). A dioxane hydrochloride solution (100 mL) was added under an ice-water bath. The mixture was allowed to react at room temperature for 60 minutes. The reaction was monitored for completion by LCMS. The reaction solution was concentrated directly, and 1 M NaOH solution was added until the pH reached 8. DCM (100 mL) was added to the reaction solution, and the organic phase was collected and washed once with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to yield 16.8 g of a white solid, compound M5-2.

[0181] ESI-MS m / z:332[M+H] + .

[0182] Step 3: Synthesis of Compound M5-3

[0183] Compound M5-2 (19.0 g), Boc-L-valine (12.5 g), and HATU (22.9 g) were dissolved in DMF (100 mL). DIEA (28.4 mL) was added and the mixture was allowed to react at room temperature for 10 minutes. The reaction was monitored for completion by LCMS. EA (100 mL) and saturated brine (100 mL) were added to the reaction solution. The organic phase was collected, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to obtain 25.0 g of a white solid, i.e., compound M5-3.

[0184] ESI-MS m / z:531[M+H] + .

[0185] Step 4: Synthesis of Compound M5-4

[0186] Compound M5-3 (25.0 g) was added to DCM (100 mL). A dioxane hydrochloride solution (100 mL) was added under an ice-water bath. The mixture was allowed to react at room temperature for 60 minutes. The reaction was monitored for completion by LCMS. The reaction solution was directly concentrated, and 1 M NaOH solution was added until the pH reached 8. DCM (100 mL) was added to the reaction solution, and the organic phase was collected and washed once with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain 17.0 g of a white solid, compound M5-4.

[0187] ESI-MS m / z:431[M+H] + .

[0188] Step 5: Synthesis of Compound M5

[0189] Compound M5-4 (17.0 g) and TEA (32.9 mL) were dissolved in ACN (100 mL) and THF (130 mL). 2-Azido-1,3-dimethylimidazolium hexafluorophosphate (22.5 g) was dissolved in ACN (30 mL) under an ice-water bath and slowly added to the system. The reaction was incubated for 3 hours, and the reaction was monitored by LCMS for completion. The reaction solution was directly concentrated, and DCM (100 mL) was added to the reaction solution. The organic phase was collected and washed once with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to obtain 15.3 g of a light yellow powder, namely compound M5.

[0190] ESI-MS m / z:457[M+H] + .

[0191] 1 H NMR (500MHz, Methanol-d4) δ8.87(s,1H),7.46-7.40(m,4H),7.94(q,J=7.0Hz,1H),4.64-4.60(m,1H),4.45-4.44(m,1H),3.75( d,J=8.0Hz,1H),3.71-3.64(m,2H),2.48(s,3H),2.26-2.15(m,2H),1.96-1.91(m,1H),1.52(d,J=7.0Hz,3H),1.09-0.98(m,6H).

[0192] Synthesis of intermediate M6

[0193] Step 1: Synthesis of compound M6-1

[0194] N-Boc-4-hydroxypiperidine (1.0 g) was added to a 25 mL single-necked flask and dissolved in THF (10 mL). NaH (300.0 mg) and 2,5-dibromopyrazine (1.18 g) were added and heated to 50°C for 5 h. The reaction solution was evaporated to dryness. Purification by column chromatography (PE:EA = 6:1) afforded 1.38 g of a pale yellow solid, compound M6-1.

[0195] Step 2: Synthesis of compound M6-2

[0196] Compound M6-1 (690.0 mg) was added to a 25 mL single-necked flask and dissolved in DMF (10 mL). Trimethylethynylsilane (283.7 mg), CuI (73.3 mg), PdCl2(PPh3)2 (135.2 mg), and DIEA (0.95 mL) were added. Under nitrogen protection, the mixture was heated to 50°C for 3 h. Saturated brine (5 mL) was added to the reaction solution, and the mixture was extracted with EA. The organic phase was dried over anhydrous sodium sulfate and the product was separated by column chromatography (PE:EA = 6:1) to obtain 522.2 mg of a light yellow solid, compound M6-2.

[0197] Step 3: Synthesis of compound M6-3

[0198] Compound M6-2 (522.2 mg) was added to a 25 mL single-necked flask and dissolved in THF (10 mL). A 1 M TBAF / THF solution (2.2 mL) was added and allowed to react at room temperature for 2 h. The reaction solution was evaporated to dryness. Purification by column chromatography (PE:EA = 4:1) afforded 378.2 mg of a pale yellow solid, compound M6-3.

[0199] Step 4: Synthesis of compound M6-4

[0200] Compound M5 (100.0 mg) was added to a 25 mL single-necked flask and dissolved in THF (2 mL). Compound M6-3 (80.2 mg), copper sulfate (35.2 mg), water (2 mL), tert-butyl alcohol (2 mL), and sodium ascorbate (160.0 mg) were then added. The mixture was allowed to react at room temperature for 0.5 h. The reaction solution was evaporated to dryness. Purification by column chromatography (DCM:MeOH = 12:1) afforded 132.3 mg of a pale yellow solid, compound M6-4.

[0201] Step 5: Synthesis of Compound M6

[0202] Compound M6-4 (132.3 mg) was added to a 25 mL single-necked flask and dissolved in HCl / dioxane (5 mL). Methanol (1 mL) was added and the mixture was allowed to react at room temperature for 1 h. The reaction solution was evaporated to dryness, and 5 mL of saturated sodium bicarbonate solution was added to the reaction solution. The pH of the solution was adjusted to 7-8, and the solution was extracted with dichloromethane and isopropanol. The organic phase was dried over anhydrous sodium sulfate to obtain 92.2 mg of a yellow solid, Compound M6.

[0203] Synthesis of intermediate M7

[0204] Step 1: Synthesis of compound M7-1

[0205] 5-Ethynyl-2-fluoropyridine (500 mg), tert-butyl piperazine-1-carboxylate (922 mg), 5 ml of acetone, and DIEA (1.6 g) were added to a 50 ml flask and heated to 60°C for 5 h. The mixture was desolvated and dissolved in EA. The mixture was washed with water and brine, dried over sodium sulfate, filtered, and the filtrate was desolvated under reduced pressure to obtain a crude product. The crude product was purified by column chromatography to obtain compound M7-1 (1.02 g).

[0206] Step 2: Synthesis of compound M7-2

[0207] Compound M7-1 (200 mg), M5 (250 mg), sodium ascorbate (226 mg), copper sulfate (108 mg), 3 ml of tert-butanol, 3 ml of THF, and 3 ml of water were added to a 50 ml flask and reacted at room temperature for 1 hour. After the reaction was detected to be complete, EA was added for extraction. The organic phase was washed with water and brine, dried over sodium sulfate, filtered, and the filtrate was decompressed to obtain a crude product, which was purified by column chromatography to obtain compound M7-2 (198 mg).

[0208] Step 3: Synthesis of Compound M7

[0209] Compound M7-2 (198 mg) and 4N HCl / dioxane (5 ml) were added to a 50 ml flask, reacted at room temperature for 1 hour, and desolventized under reduced pressure to obtain 200 mg of a crude product of compound M7.

[0210] Synthesis of intermediate M8:

[0211] Step 1: Synthesis of compound M8-1

[0212] N-tert-Butyloxycarbonyl-4-hydroxypiperidine (1.0 g) was added to a 25 mL single-necked flask and dissolved in THF (10 mL). NaH (300.0 mg) and 2,5-dibromopyrazine (1.18 g) were added and heated to 50°C for 5 h. The reaction solution was evaporated to dryness. Purification by column chromatography (PE:EA = 6:1) afforded 1.38 g of a pale yellow solid, compound M8-1.

[0213] Step 2: Synthesis of compound M8-2

[0214] Compound M8-1 (690.0 mg) was added to a 25 mL single-necked flask and dissolved in DMF (10 mL). Trimethylethynylsilane (283.7 mg), CuI (73.3 mg), PdCl2(PPh3)2 (135.2 mg), and DIPEA (0.95 mL) were added. Under nitrogen, the mixture was heated to 50°C for 3 h. Saturated brine (5 mL) was added to the reaction solution, and the mixture was extracted with EA. The organic phase was dried over anhydrous sodium sulfate and the product was isolated by column chromatography (PE:EA = 6:1) to obtain 522.2 mg of a light yellow solid, compound M8-2.

[0215] Step 3: Synthesis of Compound M8

[0216] Compound M8-2 (522.2 mg) was added to a 25 mL single-necked flask and dissolved in THF (10 mL). A 1 M solution of TBAF in tetrahydrofuran (2.2 mL) was added and allowed to react at room temperature for 2 h. The reaction solution was evaporated to dryness. Purification by column chromatography (PE:EA = 4:1) afforded 378.2 mg of a pale yellow solid, Compound M8.

[0217] Synthesis of intermediate M9

[0218] Step 1: Synthesis of compound M9-1

[0219] To a reaction flask, add 5-chloropyrazine-2-carboxaldehyde (336.23 mg), tert-butyl 3,9-diazaspiro[5.5]undecane-3-carboxylate (500.00 mg), methanol (8.00 mL), and acetic acid (0.02 mL). Stir at room temperature for 1 hour, cool to 0°C, add sodium borohydride (148.72 mg), react at room temperature for 0.5 hour, dilute with water, extract with DCM, dry the organic phase, and concentrate. Purify by column chromatography (PE:EA = 2:1) to obtain 250 mg of a yellow oil, compound M9-1.

[0220] Step 2: Synthesis of compound M9-2

[0221] Compound M9-1 (240.00 mg), trimethylsilyl acetylene (0.18 mL), bistriphenylphosphine palladium dichloride (44.22 mg), cuprous iodide (12.0 mg), triethylamine (0.53 mL), and tetrahydrofuran (5.00 mL) were added to the reaction flask. After nitrogen replacement, the mixture was reacted at 50° C. under nitrogen protection for 2 h. The reaction liquid was filtered and rinsed three times with EA. The mother liquor was concentrated and purified by column chromatography (PE:EA=1:1) to obtain 220 mg of a brown liquid, namely compound M9-2.

[0222] Step 3: Synthesis of compound M9-3

[0223] Compound M9-2 (220.00 mg), methanol (4.00 mL), and potassium carbonate (137.38 mg) were added to the reaction flask, reacted at room temperature for 0.2 h, filtered, and the filter cake was washed several times with EA. The mother liquor was concentrated and purified by column chromatography (DCM:MeOH=10:1) to obtain 50 mg of brown liquid, namely compound M9-3.

[0224] Step 4: Synthesis of Compound M9-4

[0225] Compound M9-3 (50.00 mg), M5 (61.6 mg), copper sulfate (19.39 mg), sodium ascorbate (69.54 mg), tetrahydrofuran (1.00 mL), tert-butanol (1.00 mL), and water (1.00 mL) were added to the reaction flask and reacted at room temperature for 0.5 h. The mixture was diluted with water and then extracted with DCM. The organic phase was dried, concentrated, and purified by column chromatography (DCM: ammonia methanol = 14:1) to give 65 mg of a yellow solid, namely compound M9-4.

[0226] Step 5: Synthesis of Compound M9

[0227] Compound M9-4 (65.00 mg) and dichloromethane (2.00 mL) were added to the reaction flask, and 4 M hydrogen chloride dioxane solution (0.40 mL) was added with stirring. The mixture was reacted at room temperature for 0.2 h, and then the pH was adjusted to 8 with aqueous sodium bicarbonate solution. The mixture was extracted with DCM, dried, and concentrated to obtain 45 mg of a yellow solid, namely compound M9.

[0228] Synthesis of intermediate M10:

[0229] Step 1: Synthesis of compound M10-1

[0230] Compound (R)-tert-butyl (1-(4-bromophenyl)-2-hydroxyethyl)carbamate (1.0 g), 4-methylthiazole (0.63 g), palladium acetate (0.07 g), and potassium carbonate (0.62 g) were dissolved in 10.0 mL of DMAc and reacted at 120°C for 2.0 h. The reaction solution was cooled to room temperature and quenched with water dropwise. The solution was extracted three times with ethyl acetate, and the organic phases were combined, dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The product was then purified by column chromatography to obtain 1.05 g of the product, compound M10-1.

[0231] Step 2: Synthesis of compound M10-2

[0232] Compound M10-1 (0.9 g), phthalimide (0.395 g), and triphenylphosphine (0.74 g) were dissolved in 10.0 mL of THF. The atmosphere was purged with nitrogen 2-3 times. Diethyl azodicarboxylate (0.468 g) was added under ice-cooling and allowed to react at room temperature for 1 h. The solvent was evaporated under reduced pressure to yield 1.10 g of compound M10-2, which was used directly in the next step.

[0233] Step 3: Synthesis of compound M10-3

[0234] Compound M10-2 (1.1 g) was dissolved in 10.0 mL of methanol, and hydrazine hydrate (0.74 mL) was added. The mixture was reacted at 90°C for 1.0 h. The reaction mixture was filtered to remove insoluble impurities, and the solvent was evaporated under reduced pressure to obtain 0.78 g of crude product, which was used directly in the next step to obtain compound M10-3.

[0235] Step 4: Synthesis of compound M10-4

[0236] Compound M10-3 (0.78 g) was added to a 25 mL single-necked flask and dissolved in anhydrous methanol (10 mL). Aqueous formaldehyde solution (1.9 g, 37%) was added, followed by acetic acid (0.13 mL). The mixture was allowed to react at room temperature for 30 min, followed by sodium cyanoborohydride (0.44 g). Saturated brine (5 mL) was added to the reaction solution, and the mixture was extracted with EA. The organic phase was dried over anhydrous sodium sulfate and the product was isolated by column chromatography (PE:EA = 6:1) to obtain 0.80 g of a light yellow solid, compound M10-4.

[0237] Step 5: Synthesis of Compound M10-5

[0238] Compound M10-4 (0.8 g) was dissolved in 4 M hydrogen chloride in dioxane (10.00 mL) and allowed to react at room temperature for 1.0 h. The reaction solution was evaporated under reduced pressure to remove the solvent. After re-dissolving in methanol, the solvent was again evaporated under reduced pressure to obtain 0.8 g of a yellow product, which was used directly in the next step, namely compound M10-5.

[0239] Step 6: Synthesis of Compound M10-6

[0240] Compound M10-5 (0.8 g), (2S,4R)-1-tert-butoxycarbonyl-2-carbamoyl-4-hydroxypyrrolidine (0.5 g), and DIPEA (2.3 mL) were dissolved in 10.00 mL of DCM. HATU (0.9 g) was added and the mixture was allowed to react at room temperature for 1.0 h. Silica gel was added to the reaction solution and purified by column chromatography to obtain 1.0 g of product, compound M10-6.

[0241] Step 7: Synthesis of Compound M10-7

[0242] Compound M10-6 (1.0 g) was dissolved in 4 M hydrogen chloride in dioxane (10.00 mL) and allowed to react at room temperature for 1.0 h. The reaction solution was evaporated under reduced pressure to remove the solvent. After re-dissolving in methanol, the solvent was again evaporated under reduced pressure to obtain 0.76 g of a yellow product, which was used directly in the next step, namely compound M10-7.

[0243] Step 8: Synthesis of Compound M10-8

[0244] Compound M10-7 (1.1 g), N-tert-butyloxycarbonyl-L-valine (0.4 g), and DIPEA (3.22 mL) were dissolved in 15.00 mL of DMF, and HATU (0.7 g) was added. The mixture was allowed to react at room temperature for 1.0 h. The reaction solution was quenched with water and extracted three times with ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The product was then purified by column chromatography to obtain 0.86 g of the product, compound M10-8.

[0245] Step 9: Synthesis of compound M10-9

[0246] Compound M10-8 (0.86 g) was dissolved in 4 M hydrogen chloride in dioxane (10.00 mL) and allowed to react at room temperature for 1.0 h. The reaction solution was evaporated under reduced pressure to remove the solvent. After re-dissolving in methanol, the solvent was again evaporated under reduced pressure to obtain 0.70 g of a yellow product, which was used directly in the next step, namely compound M10-9.

[0247] Step 10: Synthesis of Compound M10-10

[0248] Compound M10-9 (0.70 g) was dissolved in a mixed solvent of anhydrous acetonitrile (10.00 mL) and tetrahydrofuran (10.0 mL). Triethylamine (1.91 mL) was added, and 2-azido-1,3-dimethylimidazolium hexafluorophosphate (0.78 g) was added under ice-cooling. The mixture was allowed to react at room temperature for 2.0 h. The reaction solution was quenched with water and extracted three times with ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The product was then purified by column chromatography to obtain 0.35 g of the product, compound M10-10.

[0249] Step 11: Synthesis of Compound M10-11

[0250] M10-10 (150 mg), M8 (150 mg), sodium ascorbate (155 mg), and anhydrous copper sulfate (43 mg) were dissolved in a mixture of 2.00 mL of tert-butanol, 2.00 mL of tetrahydrofuran, and 2.00 mL of water at room temperature and allowed to react for 1 h. The reaction mixture was quenched by adding 50 mL of aqueous solution and extracted three times with a 10 / 1 DCM / MeOH mixture. The organic phases were combined, dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The mixture was then purified by column chromatography (10 / 1 DCM / MeOH) to obtain 208 mg of a yellow solid product, compound M10-11.

[0251] Step 12: Synthesis of Compound M10

[0252] Compound M10-11 (208 mg) was added to a 25 mL single-necked flask and dissolved in 4 M hydrogen chloride in dioxane (5 mL). Methanol (1 mL) was added and the mixture was allowed to react at room temperature for 1 h. The reaction solution was evaporated to dryness, and 5 mL of saturated sodium bicarbonate solution was added to the reaction solution. The pH of the solution was adjusted to 7-8, and the solution was extracted with dichloromethane / isopropanol. The organic phase was dried over anhydrous sodium sulfate to obtain 133 mg of a yellow solid, compound M10.

[0253] Example

[0254] Example 5: Synthesis of Compound (2S,4R)-1-((S)-2-(4-(5-((1-((1-((7-(8-ethyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluoro-4-(1,4-oxazepin-4-yl)pyrido[4,3-d]pyrimidin-2-yl)oxy)methyl)cyclopropyl)methyl)piperidin-4-yl)oxy)pyrazin-2-yl)-1H-1,2,3-triazol-1-yl)-3-methylbutanoyl)-4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide

[0255] Step 1: Synthesis of compound 5-1

[0256] Compound M2, 2,4,7-trichloro-8-fluoropyrido[4,3-d]pyrimidine (500.0 mg) was dissolved in dichloromethane (20 mL). Under nitrogen, the temperature was lowered to -60°C, DIPEA (690 μL) was added, and a dichloromethane solution of homomorpholine (200.0 mg) was slowly added. The temperature was maintained for 0.5 h. LCMS monitored the reaction completion. Saturated ammonium chloride (5 mL) was added to the reaction solution to quench the reaction. EA was added for extraction. The organic phase was washed once with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to obtain 610.0 mg of a white solid, i.e., compound 5-1. ESI-MS m / z: 317.1 [M+H] + .

[0257] Step 2: Synthesis of compound 5-2

[0258] Compound 5-1 (610 mg) was dissolved in DMF (20 mL), and 1,1-cyclopropane dimethanol (393 mg), Cs2CO3 (1.88 g), and DABCO (43.0 mg) were slowly added. The mixture was allowed to react at room temperature for 2 h. LCMS monitored the reaction for completion. EA was added to the reaction solution for extraction, and the mixture was washed once with saturated brine. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to obtain 757 mg of a white solid, namely compound 5-2. ESI-MS m / z: 383.2 [M+H] + .

[0259] Step 3: Synthesis of compound 5-3

[0260] Compound 5-2 (757 mg) was dissolved in DCM (20 mL). DMP reagent (1.26 g) was slowly added under ice bath conditions and the mixture was slowly warmed to room temperature for 1 h. LCMS monitored the reaction completion. DCM was added to the reaction solution for extraction, and the mixture was washed once with saturated brine. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to obtain 638 mg of a white solid, i.e., compound 5-3. ESI-MS m / z: 381.1 [M+H] + .

[0261] Step 4: Synthesis of compound 5-4

[0262] Compound 5-3 (638 mg), compound M4 (1.06 g), CataCXiumAPdG3 (244 mg), and K3PO4 (1.07 g) were weighed, 1,4-dioxane (20 mL) and water (5 mL) were added, and the mixture was reacted at 90°C for 2 h under nitrogen protection. LCMS monitored the reaction completion. EA was added to the reaction solution for extraction, and the mixture was washed once with saturated brine. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to obtain 460 mg of a white solid, namely compound 5-4. ESI-MS m / z: 535.3 [M+H] + .

[0263] Step 5: Synthesis of compound 5

[0264] Compound 5-4 (120.0 mg) and compound M6 (148 mg) were dissolved in methanol (10 mL), zinc chloride solution (0.45 mL / 1.0 M in THF) was added, and the mixture was reacted at 40°C for 1 hour. Then, sodium cyanoborohydride (42.3 mg) was added and the mixture was reacted at 60°C for 12 hours. The reaction was monitored by LCMS to be complete. Saturated sodium bicarbonate solution (5.0 mL) was added to quench the reaction, and DCM (20 mL) was added to the reaction solution. The organic phase was collected, concentrated, and subjected to preparative liquid separation to obtain 88.0 mg of compound 5. ESI-MS m / z: 590.31 / 2 [M+2H]+ .

[0265] 1 H NMR(500MHz,DMSO-d6)δ9.90(s,1H),9.17(s,1H),8.99(s,1H),8.76(d,J=1.3Hz, 1H),8.57(s,1H),8.52(d,J=7.9Hz,1H),8.29(d,J=1.3Hz,1H),7.77(dd,J=9.1,6 .0Hz,1H),7.48–7.42(m,2H),7.39–7.32(m,4H),7.03(d,J=2.5Hz,1H),5.40(d,J =10.2Hz,1H),5.18(d,J=3.8Hz,1H),5.00(d,J=16.3Hz,1H),4.93(p,J=7.0Hz,1H) ,4.44–4.30(m,4H),4.13(t,J=15.4Hz,4H),3.95(t,J=4.5Hz,2H),3.77–3.73(m, 3H),2.80(s,2H),2.46(s,2H),2.36(s,3H),2.25(s,2H),2.12–2.05(m,3H),2.02 –1.94(m,3H),1.79(ddd,J=12.8,8.5,4.5Hz,1H),1.67(d,J=9.2Hz,2H),1.39(d, J=7.0Hz,3H),1.08(d,J=6.4Hz,3H),0.72(dt,J=32.8,18.5Hz,11H),0.44(s,2H).

[0266] Example 7: Synthesis of Compound (2S,4R)-1-((S)-2-(4-(5-(1-((4-(1,1-dioxo-1,3-thiazin-3-yl)-7-(8-ethyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluoropyrido[4,3-d]pyrimidin-2-yl)oxy)methyl)cyclopropyl)methyl)piperidin-4-yl)oxy)pyrazin-2-yl)-1H-1,2,3-triazol-1-yl)-3-methylbutanoyl)-4-hydroxy-N-((S)-1-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide

[0267] Step 1: Synthesis of compound 7-1

[0268] 3-Aminopropanethiol hydrochloride (500 mg) was dissolved in 5 mL of ethanol, and then aqueous formaldehyde solution (319 mg) was added. Stir at 20°C for 12 hours. LCMS monitored the reaction completion. Concentrate under reduced pressure to obtain crude compound 7-1 (520 mg). ESI-MS m / z: 104.1 [M+H] + .

[0269] Step 2: Synthesis of compound 7-2

[0270] Compound 7-1 (240 mg) was added to a 25 mL single-necked flask, followed by 4 mL of tetrahydrofuran. M1 (450 mg) was then added. 1.18 mL of N,N-diisopropylethylamine was added dropwise to the reaction mixture at -20°C. Stir at 0°C for 2 hours. LCMS monitored the reaction completion. The mixture was concentrated under reduced pressure and purified by column chromatography to afford compound 7-2 (441 mg). ESI-MS m / z: 319.1 [M+H] + .

[0271] Step 3: Synthesis of compound 7-3

[0272] Compound 7-2 (441 mg) was added to a 25 mL single-necked flask, followed by 6 mL of dichloromethane. m-Chloroperbenzoic acid (701 mg) was then added and stirred at 20°C for 0.5 hours. LCMS monitored the reaction completion. The mixture was concentrated under reduced pressure and purified by column chromatography to afford compound 7-3 (374 mg). ESI-MS m / z: 351.1 [M+H] + .

[0273] Step 4: Synthesis of compound 7-4

[0274] Compound 7-3 (374 mg) was added to a 50 mL single-necked bottle, followed by 5 mL of N,N-dimethylformamide, 1,1-cyclopropane dimethanol (218 mg), triethylenediamine (24 mg), and cesium carbonate (694 mg). The mixture was stirred at 20°C for 12 hours. LCMS monitored the reaction completion. The reaction solution was diluted with water and extracted twice with ethyl acetate. The organic phases were collected and combined, dried over anhydrous sodium sulfate, and purified by column chromatography to obtain compound 7-4 (70 mg). ESI-MS m / z: 417.1 [M+H] + .

[0275] Step 5: Synthesis of compound 7-5

[0276] Compound 7-4 (70 mg) was added to a 10 mL single-necked vial, followed by 2 mL of dichloromethane and Dess-Martin periodinane (142 mg). The mixture was stirred at 20°C for 12 hours. LCMS monitored the reaction completion. The mixture was concentrated under reduced pressure and purified by column chromatography to afford compound 7-5 (36 mg). ESI-MS m / z: 415.1 [M+H] + .

[0277] Step 6: Synthesis of compound 7-6

[0278] Compound 7-5 (10 mg) and M4 (15 mg) were added to a 10 mL single-necked bottle, followed by 1 mL of 1,4-dioxane and 0.2 mL of water. Then, methanesulfonic acid [n-butyldi(1-adamantyl)phosphine] (2-amino-1,1'-biphenyl-2-yl) palladium (II) (4 mg) and potassium phosphate (15 mg) were added. The mixture was replaced with N2 three times and stirred at 90°C under a N2 atmosphere for 10 hours. LCMS monitored the reaction completion. The mixture was concentrated under reduced pressure and purified by column chromatography to obtain compound 7-6 (5 mg). ESI-MS m / z: 569.2 [M+H] + .

[0279] Step 7: Synthesis of compound 7

[0280] M6 (9 mg) was added to a 10 mL single-necked bottle, 0.5 mL of methanol was added, compound 7-6 (5 mg) was added, 0.01 mL of zinc chloride (1 mol / L THF) was added, and the mixture was stirred at 40 ° C for 2 hours. Sodium cyanoborohydride (3 mg) was added and stirred at 50 ° C for 12 hours. LCMS monitoring of the reaction was complete. The reaction was quenched with saturated ammonium chloride solution, and the reaction solution was neutralized with saturated sodium bicarbonate solution. The reaction solution was extracted twice with dichloromethane / methanol (5:1), and the organic phases were collected and combined, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by pre-HPLC to obtain compound 7 (1.7 mg). ESI-MS m / z: 607.07 1 / 2 [M+2H] + .

[0281] Example 57: Synthesis of Compound (2S,4R)-1-((2S)-2-(4-(5-((1-((1-((1-(4-(3-(cyanomethyl)-3-hydroxypiperidin-1-yl)-7-(8-ethyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluoropyrido[4,3-d]pyrimidin-2-yl)oxy)methyl)cyclopropyl)methyl)piperidin-4-yl)oxy)pyrazin-2-yl)-1H-1,2,3-triazol-1-yl)-3-methylbutanoyl)-4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide

[0282] Step 1: Synthesis of compound 57-1

[0283] Compound M2 (1.2 g), 1,1-cyclopropane dimethanol (0.7 g), cesium carbonate (2.2 g), and DABCO (0.1 g) were dissolved in DMF (10 mL) and stirred at room temperature overnight. After the reaction was complete, the mixture was diluted with water and extracted twice with EA. The mixture was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and dried by spin drying. The mixture was then purified by column chromatography (PE:EA = 100:67) to obtain a racemate (0.8 g). ESI-MS m / z: 422.1 [M+H] + The product was separated by liquid chromatography using the following conditions: CHIRAL ART IA column (20.0 mm × 250 mm, 5 μm); column temperature: 35°C; wavelength: 254 nm; flow rate: 25 ml / min; retention time: 4.217 min; mobile phase: 0.1% diethylamine in n-hexane: 0.1% diethylamine in ethanol (40:60), to obtain chiral compound 57-1.

[0284] Step 2: Synthesis of compound 57-2

[0285] Compound 57-1 (130.0 mg), compound M4 (2-(8-ethyl-7-fluoro-3-methoxymethoxy)naphthalen-1-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane) (229.7 mg), CataCXiumAPdG3 (46.5 mg), K3PO4 (203.1 mg), 1,4-dioxane (4 mL), and H2O (1 mL) were heated to 90°C and stirred for 1 h. After the reaction was complete, the mixture was cooled to room temperature, diluted with water, extracted twice with EA, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, dried, and purified by column chromatography (DCM:MeOH = 100:7) to obtain compound 57-2 (185 mg). ESI-MS m / z: 620.3 [M+H] + .

[0286] Step 3: Synthesis of compound 57-3

[0287] To a solution of compound 57-2 (185 mg) in DCM (10 mL) was added DMP (259.0 mg) and stirred at room temperature for 30 min. After the reaction was complete, the mixture was diluted with water and extracted twice with DCM. The combined organic phases were dried over anhydrous sodium sulfate, filtered, and dried by spin chromatography (PE:EA = 50:50) to obtain compound 57-3 (110 mg). ESI-MS m / z: 618.4 [M+H] + .

[0288] Step 4: Synthesis of compound 57-4

[0289] To a solution of compound 57-3 (80 mg) in methanol (10 mL) were added M6 (87 mg) and ZnCl2 (0.6 mL), and the mixture was heated to 40°C and stirred for 1 h. Subsequently, sodium cyanoborohydride (33 mg) was added, and the mixture was heated to 70°C and stirred for 4 h. After the reaction was complete, the solvent was evaporated and the mixture was purified by column chromatography (DCM:MeOH = 100:13) to obtain compound 57-4 (100 mg). ESI-MS m / z: 631.74 [M / 2+H] + .

[0290] Step 5: Synthesis of compound 57

[0291] To a solution of compound 57-4 (100 mg) in DCM (8 mL) was slowly added trifluoroacetic acid (2 mL) and stirred at room temperature for 20 min. After the reaction was complete, the solvent was evaporated and the concentrate was subjected to preparative liquid chromatography (chromatographic column: Kinetex EVO C18 (21×150 mm, 5 μm); mobile phase: ACN + 0.05% TFA (20%-53%; retention time: 9.1-9.65 min) to obtain compound 57 (26 mg). ESI-MS m / z: 609.72 1 / 2 [M+2H] + .

[0292] 1 H NMR (500 MHz, DMSO-d6) 1H NMR (500MHz, DMSO) δ9.20 (s, 1H), 8.99 (d, J = 4.4Hz, 1H), 8.76 (s, 1H), 8.59 –8.49(m,2H),8.30(s,1H),7.80–7.73(m,1H),7.44(d,J=8.1Hz,2H),7.35 (dd,J=12.8,8.2Hz,4H),7.03(s,1H),5.54(s,1H),5.40(d,J=10.0Hz,1H) ,5.18(d,J=3.6Hz,1H),5.02(s,1H),4.95–4.90(m,1H),4.40(t,J=8.1Hz, 1H),4.35–4.27(m,3H),4.11(dd,J=28.6,13.1Hz,1H),3.74(dd,J=41.9,1 3.7Hz,3H),3.50–3.40(m,2H),2.81(s,4H),2.37(s,3H),2.25(s,3H),2.0 8–1.95(m,5H),1.76(dd,J=47.7,31.5Hz,8H),1.55(d,J=6.6Hz,1H),1.39 (dt,J=22.5,11.3Hz,5H),1.24(s,6H),1.08(d,J=6.3Hz,4H),0.45(s,2H).

[0293] Example 111: Synthesis of Compound (2S,4R)-1-((2S)-2-(4-(6-(4-(1-(((7-(8-ethyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluoro-4-(1-oxa-6-azaspiro[3.5]non-6-yl)pyrido[4,3-d]pyrimidin-2-yl)oxy)methyl)cyclopropyl)methyl)piperazin-1-yl)pyridin-3-yl)-1H-1-1,2,3-triazol-1-yl)-3-methylbutanoyl)-4-hydroxy-N-(((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide

[0294] Step 1: Synthesis of compound 111-1

[0295] 1-Oxa-6-azaspiro[3.5]nonane (185 mg), 5 ml of DCM, and DIEA (280 mg) were added to a 50 ml flask, the temperature was lowered to -40°C, compound M1 (250 mg) was added, and the mixture was kept warm for 1 hour. DCM was added to dissolve the mixture, washed with water and brine, dried over sodium sulfate, filtered, and the filtrate was decompressed to obtain a crude product, which was purified by column chromatography to obtain compound 111-1 (250 mg).

[0296] Step 2: Synthesis of compound 111-2

[0297] Compound 111-1 (250 mg), cyclopropane-1,1-diyldimethanol (149 mg), 5 ml of DMF, DABCO (15 mg), and cesium carbonate (400 mg) were added to a 50 ml flask and reacted at room temperature for 6 hours. EA was added to dissolve the mixture, washed with water and brine, dried over sodium sulfate, filtered, and the filtrate was desolvated under reduced pressure to obtain a crude product, which was purified by column chromatography to obtain compound 111-2 (225 mg).

[0298] Step 3: Synthesis of compound 111-3

[0299] Compound 111-2 (225 mg), M3 (300 mg), 2 ml of dioxane, 0.2 ml of water, potassium phosphate (350 mg), and CataCXium APd G3 (40 mg) were added to a 50 ml flask, replaced with nitrogen and protected, heated to 90 ° C, reacted for 2 hours, dissolved in EA, washed with water, washed with brine, dried over sodium sulfate, filtered, and the filtrate was decompressed to obtain a crude product, which was purified by column chromatography to obtain compound 111-3 (229 mg).

[0300] Step 4: Synthesis of compound 111-4

[0301] Compound 111-3 (100 mg), 5 ml of DCM, and DMP (84 mg) were added to a 50 ml flask and reacted at room temperature for 1 hour. The mixture was dissolved in DCM, washed with water and brine, dried over sodium sulfate, filtered, and the filtrate was desolvated under reduced pressure to obtain a crude product, which was purified by column chromatography to obtain compound 111-4 (95 mg).

[0302] Step 5: Synthesis of compound 111-5

[0303] Compound 111-4 (95 mg), M7 (100 mg), MeOH 5 ml, and 1N ZnCl2 solution 0.3 ul were added to a 50 ml flask, and the mixture was heated to 45 °C for 1 hour. Sodium cyanoborohydride (100 mg) was added and the reaction was continued at 45 °C for 12 hours. The mixture was diluted with DCM, washed with water and brine, dried over sodium sulfate, filtered, and the filtrate was decompressed to obtain a crude product, which was purified by column chromatography to obtain compound 111-5 (60 mg).

[0304] Step 6: Synthesis of compound 111

[0305] Compound 111-5 (60 mg), 3 ml of DCM, and 1 ml of TFA were added to a 50 ml flask and reacted at room temperature for 0.5 hours. After detection, the mixture was desolvated under reduced pressure, dissolved in DCM, and adjusted to pH 7-8 with a saturated aqueous solution of baking soda. The mixture was stirred, separated, washed with brine, dried over sodium sulfate, and filtered. The filtrate was desolvated under reduced pressure to obtain the crude product, which was then purified by Pre-HPLC to obtain compound 111 (25.0 mg). ESI-MS m / z: 595.27 1 / 2 [M+2H] +

[0306] 1 H NMR(500MHz,DMSO-d6)ppm 9.94(s,1H),9.21(d,J=4.09Hz,1H),8.99(s,1H),8.68-8.46(m,1H),8.00(dd,J=8.76,1.96Hz,1H),7.77(dd,J =9.12,6.02Hz,1H),7.52-7.17(m,1H),7.06(dd,J=4.46,2.62Hz,1H),6.88(d,J=8.85Hz,1H),5.32(d,J=10.26 Hz,1H),5.19(d,J=3.72Hz,1H),5.04-4.80(m,1H),4.62-4.08(m,1H),3.98-3.26(m,1H),3.25-3.17(m,1H),2. 63-1.97(m,1H),1.96-1.63(m,1H),1.56(d,J=6.98Hz,1H),1.47-1.17(m,1H),1.16-0.60(m,1H),0.46(s,1H).

[0307] Example 112: Synthesis of Compound (2S,4R)-1-((S)-2-(4-(5-(1-((((S)-2-(8-ethyl-7-fluoro-3-hydroxynaphthalen-1-yl)-1-fluoro-5a,6,9,10-tetrahydro-5H,8H-4,7-dioxa-3,10a,11,13-tetraazanaphtho[1,8-ab]hepten-12-yl)oxy)methyl)cyclopropyl)methyl)piperidin-4-yl)oxy)pyrazin-2-yl)-1H-1,2,3-triazol-1-yl)-3-methylbutanoyl)-4-hydroxy N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide

[0308] Step 1: Synthesis of compound 112-1

[0309] To a reaction flask, 7,9-dichloro-10-fluoro-3-methylsulfanyl-2,4,8-triazabicyclo[4.4.0]dec-2,4,6,8,10-penten-5-ol (400.00 mg), tetrahydrofuran (8.00 mL), and sodium hydride (102.82 mg) were added. The mixture was allowed to react at room temperature for 5 minutes, followed by the addition of [(3R)-1,4-oxolan-3-yl]methanol (196.68 mg) and the reaction was continued at room temperature for 1 hour. The reaction was quenched with aqueous ammonium chloride, and the precipitated solid was filtered. The filter cake was washed three times with water and once with EA, and finally dried to yield 450 mg of a yellow solid, compound 112-1.

[0310] Step 2: Synthesis of compound 112-2

[0311] Compound 112-1 (440.00 mg), N,N-dimethylformamide (6.00 mL), and N,N-diisopropylethylamine (0.58 mL) were added to a reaction flask. HATU (670.00 mg) was added with stirring and the mixture was allowed to react at room temperature for 0.5 h. Water was added to precipitate a solid, which was filtered and washed three times with water before drying to yield 310 mg of a yellow solid, compound 112-2.

[0312] Step 3: Synthesis of compound 112-3

[0313] To a reaction flask were added compound 112-2 (300.00 mg), 2-[8-ethyl-7-fluoro-3-(methoxymethoxy)naphthalen-1-yl]-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (605.76 mg), [n-butyldi(1-adamantyl)phosphine](2-amino-1,1'-biphenyl-2-yl)palladium(II) methanesulfonate (91.83 mg), potassium phosphate (535.42 mg), 1,4-dioxane (6.00 mL), and water (1.00 mL). The mixture was reacted at 90°C under nitrogen for 2 h. The mixture was diluted with water and extracted with EA. The organic phase was dried and concentrated. Purification by column chromatography (DCM:EA = 5:1) afforded 410 mg of a yellow solid, compound 112-3.

[0314] Step 4: Synthesis of compound 112-4

[0315] Compound 112-3 (410.00 mg), dichloromethane (6.00 mL), and m-chloroperbenzoic acid (229.64 mg) were added to a reaction flask and reacted at room temperature for 0.5 h. Aqueous sodium bicarbonate solution was added to adjust the pH to 8. The mixture was then extracted with DCM, and the organic phase was dried and concentrated to obtain 415 mg of a crude yellow solid, compound 112-4.

[0316] Step 5: Synthesis of compound 112-5

[0317] To a reaction flask, [1-(Hydroxymethyl)cyclopropyl]methanol (114.55 mg), tetrahydrofuran (6.00 mL), compound 112-4 (320.00 mg), and sodium tert-butoxide (107.79 mg) were added. The mixture was reacted at room temperature for 10 min. The reaction was quenched with aqueous ammonium chloride under an ice bath, and then extracted with EA. The organic phase was dried and concentrated. Purification by column chromatography (DCM:EA = 3:1) afforded 180 mg of a yellow solid, compound 112-5.

[0318] Step 6: Synthesis of compound 112-6

[0319] Compound 112-5 (170.00 mg), dichloromethane (5.00 mL), and Dess-Martin periodinane (154.00 mg) were added to a reaction flask and reacted at room temperature for 0.5 h. The mixture was diluted with saturated aqueous sodium bicarbonate solution and extracted with DCM. The organic phase was dried and concentrated. Purification by column chromatography (DCM:EA = 3:1) afforded 95 mg of a yellow solid, compound 112-6.

[0320] Step 7: Synthesis of compound 112-7

[0321] Compound 112-6 (45.00 mg), intermediate M6 (53.84 mg), methanol (1.50 mL), 1,2-dichloroethane (1.50 mL), and 1 M zinc chloride in tetrahydrofuran (0.07 mL) were added to a reaction flask and reacted at 40°C for 1 h. Sodium cyanoborohydride (37.30 mg) was then added and reacted at 60°C for 3 h. The mixture was diluted with water and extracted with DCM and MeOH. The organic phase was dried and concentrated. Pre-TLC purification afforded 52 mg of a light yellow solid, compound 112-7.

[0322] Step 8: Synthesis of compound 112

[0323] Compound 112-7 (45.00 mg), dichloromethane (1.00 mL), and trifluoroacetic acid (1.00 mL) were added to a reaction flask and allowed to react at room temperature for 0.5 h. The reaction solution was concentrated and dissolved in DCM. The reaction solution was then added to an icy sodium bicarbonate aqueous solution and adjusted to pH 8. The solution was then extracted with DCM and methanol. The organic phase was dried and concentrated. Pre-TLC purification afforded compound 112 (31 mg). LCMS: 1 / 2 [M+2H] + =604.21

[0324] 1H NMR (500MHz, Methanol-d4) δ8.86(d,J=9.1Hz,1H),8.71(t,J=1.3Hz,1H),8.55(d,J=13.8Hz,1H),8.17(dd,J=4.7,1.3Hz,1H),7.66–7.59(m,1H),7.47–7. 37(m,4H),7.30–7.18(m,2H),7.10(dd,J=48.6,2.3Hz,1H),5.40(d,J=10.1H z,1H),5.20(m,1H),5.06(m,2H),4.76–4.68(m,1H),4.63–4.52(m,2H),4.49– 4.41(m,3H),4.37–4.25(m,1H),4.16(m,1H),4.01–3.85(m,3H),3.76(m,1H) ,3.60–3.53(m,1H),3.48–3.36(m,1H),3.31(m,3H),3.00(d,J=10.8Hz,2H),2 .70–2.48(m,5H),2.44–2.27(m,3H),2.25–2.06(m,4H),1.94(m,4H),1.52(d ,J=7.0Hz,3H),1.15(t,J=6.9Hz,3H),0.86(m,6H),0.76(s,2H),0.57(s,2H).

[0325] Example 113: Synthesis of Compound (2S,4R)-1-((S)-2-(4-(5-(1-((1-((7-(8-ethyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluoro-4-(3-oxoazepin-1-yl)pyrido[4,3-d]pyrimidin-2-yl)oxy)methyl)cyclopropyl)methyl)piperidin-4-yl)oxy)pyrazin-2-yl)-1H-1,2,3-triazol-1-yl)-3-methylbutanoyl)-4-hydroxy-N-((S)-1-((4-(4-methylthiazol-5-yl)phenyl)ethyl)-2-carboxamide

[0326] Step 1: Synthesis of compound 113-2

[0327] Compound 113-1 (500.0 mg) was added to a 100 mL single-necked flask, dissolved in DCM (5 mL), and TFA (2.5 mL) was added dropwise. The mixture was allowed to react at room temperature for 1 h. The reaction solution was evaporated to dryness to obtain 450 mg of a light yellow oil, which was compound 113-2.

[0328] Step 2: Synthesis of compound 113-3

[0329] Compound M1 (709.0 mg) was added to a 100 mL single-necked flask and dissolved in THF (10 mL). DIEA (1.94 mL) was then added. The resulting pale yellow oil, Compound 113-2, was dissolved in THF (2 mL) and added dropwise to the reaction mixture in an ice bath. The reaction mixture was allowed to react for 1 h. The reaction mixture was added to 30 mL of water and extracted twice with EA. The organic phases were combined, washed once with saturated brine, dried over anhydrous sodium sulfate, and purified by column chromatography (DCM:MeOH = 32:1) to yield 510 mg of a pale yellow solid, Compound 113-3.

[0330] Step 3: Synthesis of compound 113-4

[0331] Compound 113-3 (510 mg) and ethylene glycol (200 mg) were added to a 100 mL single-necked flask and dissolved in toluene (15 mL). p-Toluenesulfonic acid (26.7 mg) was added and the mixture was heated to 110°C for 5 h. After cooling, the reaction solution was concentrated by adding silica gel and purified by column chromatography (DCM:MeOH = 50:1) to obtain 175 mg of a white solid, compound 113-4.

[0332] Step 4: Synthesis of compound 113-5

[0333] Compound 113-4 (175 mg), cyclopropane dimethanol (95.4 mg), DABCO (10.5 mg), and cesium carbonate (305.4 mg) were added to a 50 mL single-necked flask and dissolved in DMF (8 mL). The mixture was stirred at room temperature for 1 h. The reaction solution was added to water (50 mL) and extracted twice with EA. The organic phases were combined, washed once with saturated brine, dried over anhydrous sodium sulfate, and purified by column chromatography (DCM:MeOH = 32:1) to obtain 210 mg of a yellow oil, compound 113-5.

[0334] Step 5: Synthesis of compound 113-6

[0335] Compound 113-5 (210 mg) was added to a 50 mL single-necked flask and dissolved in DCM (6 mL). DMP (250 mg) was added and stirred at room temperature for 1 h. The reaction solution was filtered through a filter, and the filtrate was washed once with saturated sodium bicarbonate solution and once with saturated brine, dried over anhydrous sodium sulfate, and purified by column chromatography (DCM:MeOH = 25:1) to obtain 170 mg of a yellow oil, compound 113-6.

[0336] Step 6: Synthesis of compound 113-7

[0337] To a 50 mL three-necked flask, compound 113-6 (130 mg), intermediate M4 (188 mg), CataCXium APd G3 (21.7 mg), and potassium phosphate (190 mg) were added and dissolved in 1,4-dioxane (7 mL). Water (0.7 mL) was added, and the atmosphere was purged with nitrogen three times. The mixture was stirred at 100°C for 3 h. The reaction solution was cooled to room temperature and purified by column chromatography (DCM:MeOH = 20:1) to obtain 90 mg of a yellow solid, compound 113-7.

[0338] Step 7: Synthesis of compound 113-8

[0339] Compound 113-7 (90 mg) was added to a 25 mL single-necked flask and dissolved in MeOH (5 mL). M6 (120 mg) and 1 M zinc chloride in tetrahydrofuran (0.1 mL) were added and heated to 40°C for 2 h. NaBH3CN (50 mg) was added and allowed to react overnight. The reaction solution was added to an appropriate amount of brine and extracted twice with a 5:1 solution of DCM:MeOH. The organic phases were combined, washed once with saturated brine, dried over anhydrous sodium sulfate, and purified by column chromatography (DCM:MeOH = 10:1) to obtain 95 mg of a tan solid, compound 113-8.

[0340] Step 8: Synthesis of compound 113

[0341] Compound 113-8 (95 mg) was added to a 25 mL single-necked flask and dissolved in 1,4-dioxane (2 mL). Water (0.4 mL) and 4 M dioxane hydrochloride (0.4 mL) were added and the mixture was allowed to react for 1 h. Saturated sodium bicarbonate solution was added to the reaction mixture to adjust the pH to 8. The mixture was then extracted twice with DCM:MeOH = 5:1. The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated. The product was separated by pre-HPLC (CH3CN / H2O = 60%:40%) to obtain compound 113 (19.5 mg, 98.02% purity). LCMS: 1 / 2 [M+2H] + =596.2.

[0342] Example 114: Synthesis of compound (2S,4R)-1-((2S)-2-(4-(5-((1-((1-((7-(8-ethyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluoro-4-((S)-6-hydroxy-6-methyl-1,4-oxazepan-4-yl)pyrido[4,3-d]pyrimidin-2-yl)oxy)methyl)cyclopropyl)methyl)piperidin-4-yl)oxy)pyrazin-2-yl)-1H-1,2,3-triazol-1-yl)-3-methylbutanoyl)-4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide.

[0343] Step 1: Synthesis of compound 114-1

[0344] To a 100 mL single-necked flask, (S)-6-methyl-1,4-oxazepan-6-ol (390 mg) and DIPEA (1.1 g) were added sequentially and dissolved in DCM (30 mL). The mixture was cooled to below -40°C and compound M1 (670 mg) was added portionwise. The mixture was stirred for 1.0 h. After the reaction was complete, the mixture was quenched with saturated ammonium chloride, extracted with dichloromethane, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and dried by spin drying. The mixture was then purified by column chromatography (PE:EA = 1:1) to afford a pale yellow solid, compound 114-1 (800 mg). ESI-MS m / z: 347.1 [M+H] + .

[0345] Step 2: Synthesis of compound 114-2

[0346] Compound 114-1 (400 mg), 1,1-cyclopropane dimethanol (240 mg), cesium carbonate (750 mg), and DABCO (30 mg) were added sequentially to a 50 mL single-necked flask and dissolved in DMF (15 mL). The mixture was stirred at room temperature overnight. After the reaction was complete, the mixture was diluted with water, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and dried by spin drying. The mixture was then purified by column chromatography (DCM:EA = 9:1) to afford a light yellow solid, compound 114-2 (250 mg). ESI-MS m / z: 413.2 [M+H] + .

[0347] Step 3: Synthesis of compound 114-3

[0348] To a solution of compound 114-2 (250 mg) in DCM (10 mL) was added DMP (379 mg) and stirred at room temperature for 30 min. After the reaction was complete, the mixture was diluted with water and extracted with dichloromethane. The combined organic phases were dried over anhydrous sodium sulfate, filtered, and dried by spin chromatography (DCM:EA = 6:1) to afford a light yellow solid, compound 114-3 (210 mg). ESI-MS m / z: 410.2 [M+H] + .

[0349] Step 4: Synthesis of compound 114-4

[0350] Compound 114-3 (210 mg), 2-(8-ethyl-7-fluoro-3-methoxymethoxy)naphthalen-1-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (310.7 mg), CataCXium APd G3 (70.5 mg), K3PO4 (208.1 mg), 1,4-dioxane (8 mL), and H2O (2 mL) were heated to 90°C and stirred for 1.0 h. After the reaction was complete, the mixture was cooled to room temperature and extracted with ethyl acetate. The mixture was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and dried by spin drying. The mixture was purified by column chromatography (DCM:MeOH = 15:1) to afford a pale yellow solid, compound 114-4 (150 mg). ESI-MS m / z: 565.4 [M+H] + .

[0351] Step 5: Synthesis of target compound 114

[0352] To a solution of compound 114-4 (150 mg) in methanol (40 mL) were added M6 (128 mg) and ZnCl2 (0.8 mL), and the mixture was heated to 40°C and stirred for 1.0 h. Subsequently, sodium cyanoborohydride (58 mg) was added, and the mixture was heated to 70°C and stirred for 4.0 h. After the reaction was complete, the solvent was evaporated, and the concentrate was purified by Pre-HPLC (C18 column, A: 0.05% aqueous diethylamine, B: acetonitrile, concentration gradient: 20% B to 85% B, 10.41 min, flow rate: 20 mL / min, 254 nm) to obtain the target compound 114 (24.5 mg). ESI-MS m / z: 604.72 1 / 2 [M+2H] + .

[0353] 1H NMR(500MHz,DMSO-d6)δ9.96(s,1H),9.48(s,1H),8.99(s,1H),8.76(t,J=2.4Hz,1H) ,8.59–8.50(m,2H),8.30(s,1H),7.77(dd,J=9.0,6.1Hz,1H),7.45(d,J=8.2Hz,2H), 7.35(dd,J=11.5,9.1Hz,4H),7.03(dd,J=18.0,2.4Hz,1H),5.40(d,J=10.1Hz,1H),5.18(dd,J=10.0,6.2Hz,2H),5.02(s,1H ),4.97–4.89(m,1H),4.36(dt,J=30.0,11.3Hz,6H),4.15(dd,J=29.2,14.7Hz,1H),4.08–3.94(m,3H),3.74(dd,J=34.7,9.1 Hz,2H),3.62–3.51(m,2H),2.33(t,J=38.0Hz,6H),2.19–1.93(m,6H),1.82–1.73(m,1H),1.67(s,2H),1.60–1.43(m,1H),1. 38(d,J=7.0Hz,3H),1.24(s,3H),1.10(ddd,J=22.3,18.5,7.1Hz,8H),0.85(t,J=6.9Hz,1H),0.78–0.63(m,9H),0.44(s,2H).

[0354] Example 115: Synthesis of Compound (2S,4R)-1-((S)-2-(4-(5-((9-((1-((7-(8-ethyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluoro-4-((S)-6-hydroxy-6-methyl-1,4-oxazacyclopentyl-4-yl)pyrido[4,3-d]pyrimidin-2-yl)oxy)methyl)-3,9-diazaspiro[5.5]undec-3-yl)methyl)pyrazin-2-yl)-1H-1,2,3-triazol-1-yl)-3-methylbutanoyl)-4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide

[0355] Step 1: Synthesis of compound 115-1

[0356] To a 100 mL single-necked flask, (S)-6-methyl-1,4-oxazepan-6-ol (390 mg) and DIPEA (1.1 g) were added sequentially and dissolved in DCM (30 mL). The mixture was cooled to below -40°C and compound M1 (670 mg) was added portionwise. The mixture was stirred for 1.0 h. After the reaction was complete, the mixture was quenched with saturated ammonium chloride, extracted with dichloromethane, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and dried by spin drying. The mixture was then purified by column chromatography (PE:EA = 1:1) to afford a pale yellow solid, compound 115-1 (800 mg). ESI-MS m / z: 347.1 [M+H] + Step 2: Synthesis of compound 115-2

[0357] Compound 115-1 (400 mg), 1,1-cyclopropane dimethanol (240 mg), cesium carbonate (750 mg), and DABCO (30 mg) were added sequentially to a 50 mL single-necked flask and dissolved in DMF (15 mL). The mixture was stirred at room temperature overnight. After the reaction was complete, the mixture was diluted with water, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and dried by spin drying. The mixture was purified by column chromatography (DCM:EA = 9:1) to afford a light yellow solid, compound 115-2 (250 mg). ESI-MS m / z: 413.2 [M+H] + .

[0358] Step 3: Synthesis of compound 115-3

[0359] To a solution of compound 115-2 (250 mg) in DCM (10 mL) was added DMP (379 mg) and stirred at room temperature for 30 min. After the reaction was complete, the mixture was diluted with water and extracted with dichloromethane. The combined organic phases were dried over anhydrous sodium sulfate, filtered, and dried by spin chromatography (DCM:EA = 6:1) to afford a light yellow solid, compound 115-3 (210 mg). ESI-MS m / z: 410.2 [M+H] + Step 4: Synthesis of compound 115-4

[0360] Compound 115-3 (210 mg), 2-(8-ethyl-7-fluoro-3-methoxymethoxy)naphthalen-1-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (310.7 mg), CataCXium A Pd G3 (70.5 mg), K3PO4 (208.1 mg), 1,4-dioxane (8 mL), and H2O (2 mL) were heated to 90°C and stirred for 1.0 h. After the reaction was complete, the mixture was cooled to room temperature and extracted with ethyl acetate. The mixture was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and dried by spin drying. The mixture was purified by column chromatography (DCM:MeOH = 15:1) to afford a pale yellow solid, compound 115-4 (150 mg). ESI-MS m / z: 565.4 [M+H]+ .

[0361] Step 5: Synthesis of target compound 115

[0362] To a solution of compound 115-4 (45 mg) in methanol (10 mL) were added M9 (58 mg) and ZnCl2 (0.2 mL). The mixture was heated to 40°C and stirred for 1.0 h. Subsequently, sodium cyanoborohydride (20 mg) was added and the mixture was heated to 60°C and stirred for 4.0 h. After the reaction was complete, the solvent was evaporated, and the residue was extracted with appropriate amounts of water and DCM / MeOH (10:1). The organic phase was collected, dried over anhydrous sodium sulfate, filtered, and dried. The concentrate was purified by Pre-HPLC (C18 column, A: 0.05% aqueous diethylamine, B: acetonitrile, concentration gradient: 30% B to 80% B over 9.0 min, flow rate: 16 mL / min, 254 nm) to obtain the target compound 115 (11.6 mg). ESI-MS m / z: 638.3 1 / 2 [M+2H] + .

[0363] 1 H NMR(500MHz,MeOD)δ9.50(d,J=10.4Hz,1H),9.18(t,J=4.6Hz,1H),8.85(d,J=8.5Hz,1H),8.72(d,J=11.5Hz,1H),8.66(s,1H),7.63(dd,J=9.0,5.9 Hz,1H),7.46–7.35(m,4H),7.27(d,J=2.0Hz,1H),7.21(t,J=9.3Hz,1H), 7.05(dd,J=20.2,2.4Hz,1H),5.43(d,J=10.0Hz,1H),5.09–5.01(m,1H),4 .57–4.32(m,6H),4.19–4.11(m,1H),4.03–3.94(m,2H),3.93–3.79(m,3H ),3.75–3.58(m,4H),2.68–2.58(m,1H),2.52–2.34(m,14H),2.17(dt,J=1 8.5,9.3Hz,2H),2.00–1.92(m,1H),1.56–1.39(m,11H),1.26(d,J=15.6Hz ,3H),1.15(t,J=7.8Hz,3H),0.86–0.75(m,6H),0.68(s,2H),0.47(s,2H).

[0364] Example 131: Synthesis of Compound (2S,4R)-N-((R)-2-(dimethylamino)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)-1-(S)-2-(4-(5-(1-)(1-(((7-(8-ethyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluoro-4-((S)-6-hydroxy-6-methyl-1,4-oxol-4-yl)pyrido[4,3-d]pyrimidin-2-yl)oxy)methyl)cyclopropyl)methyl)piperidin-4-yl)oxy)pyrazin-2-yl)-1H-1,2,3-triazol-1-yl)-3-methylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide

[0365] To 115-4 (36.13 mg) and intermediate M10 (45.00 mg) in methanol (1.50 mL) was added 1 M zinc chloride solution in tetrahydrofuran (0.06 mL), and the mixture was reacted at 50°C for 1 h. Sodium cyanoborohydride (16.09 mg) was then added, and the mixture was reacted at 50°C for 6 h. The reaction mixture was added to 30 mL of water and extracted with DCM. The organic phase was dried, filtered, and concentrated under reduced pressure. The residue was purified by Pre-TLC (MeOH:DCM = 1:12) to afford 28.90 mg of compound 131 (95.64% purity). LCMS: 1 / 2 [M+2H] + =626.20

[0366] 1 H NMR(500MHz, Methanol-d4)δ9.43(s,1H),8.79(s,1H),8.64(s,1H),8.45(s,1H),8.09(d,J=5.0Hz,1H),7.59–7.56(m,1H),7.39–7.29(m,4 H),7.20(s,1H),7.23(t,J=10.0Hz,1H),6.95(s,1H),5.32–5.30(m,1H),5.06–5.02(m,3H),4.55–4.44(m,3H),4.37(d,J=5.0Hz,4H),4.12 –4.08(m,1H),3.95–3.90(m,2H),3.85–3.76(m,4H),3.65–3.55(m,3H),2.88–2.84(m,3H),2.54–2.48(m,4H),2.39–2.36(m,4H),2.13–2.0 8(m,3H),1.98–1.94(m,3H),1.88–1.83(m,2H),1.71(d,J=5.0Hz,3H),1.06(d,J=5.0Hz,3H),0.74–0.72(m,7H),0.66(s,2H),0.46(s,2H).

[0367] Example 133: Synthesis of Compound (2S,4R)-1-((2S)-2-(4-(5-(1-((2-(8-ethyl-7-fluoro-3-hydroxynaphthalen-1-yl)-1-fluoro-6-hydroxy-6-methyl-5,5a,6,7,8,9-hexahydro-4-oxo-3,9a,10,12-tetraazabenzo[4,5]cycloheptyl[1,2,3-de]naphthalen-11-yl)oxy)methyl)cyclopropyl)methyl)piperidin-4-yl)oxy)pyrazin-2-yl)-1H-1,2,3-triazol-1-yl)-3-methylbutanoyl)-4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide

[0368] Step 1: Synthesis of compound 133-1

[0369] To a reaction flask, tert-butylethyl 3-oxopiperidine-1,2-dicarboxylate (2.20 g), tetrahydrofuran (6.00 mL), and methanol (1.50 mL) were added, followed by the slow addition of sodium borohydride (1.84 g). The mixture was allowed to react at room temperature for 1 h. The reaction was quenched by adding water under an ice bath, followed by extraction with EA. The organic phase was then washed once with saturated brine, dried, and concentrated. Purification by column chromatography (PE:EA = 4:1) afforded 1.74 g of a colorless liquid, compound 133-1.

[0370] Step 2: Synthesis of compound 133-2

[0371] Compound 133-1 (1.72 g), N,N-dimethylformamide (15.00 mL), and tert-butyldimethylsilyl chloride (1.46 g) were added to a reaction flask. Imidazole (1.01 g) was added with stirring and allowed to react at room temperature for 2 h. The mixture was diluted with water, extracted with EA, dried, and concentrated. Purification by column chromatography (PE:EA = 10:1) afforded 1.79 g of a colorless liquid, compound 133-2.

[0372] Step 3: Synthesis of compound 133-3

[0373] Compound 133-2 (1.79 g), dichloromethane (1.00 mL), and Dess-Martin periodinane (3.30 g) were added to a reaction flask and reacted at room temperature for 0.5 h. The mixture was diluted with saturated aqueous sodium bicarbonate, extracted with DCM, dried, and concentrated. Purification by column chromatography (PE:EA = 10:1) afforded 1.5 g of a colorless liquid, compound 133-3.

[0374] Step 4: Synthesis of compound 133-4

[0375] Compound 133-3 (1.50 g) and tetrahydrofuran (20.00 mL) were added to a reaction flask. After nitrogen replacement, the temperature was lowered to 0°C, and methylmagnesium bromide (1.04 g, 8.73 mmol) was slowly added. The reaction was allowed to return to room temperature for 10 h. The reaction was quenched with aqueous ammonium chloride in an ice bath, extracted with EA, dried, and concentrated. Purification by column chromatography (PE:EA = 3:1) afforded 0.79 g of a yellow liquid, compound 133-4.

[0376] Step 5: Synthesis of compound 133-5

[0377] Compound 133-4 (720.00 mg), dichloromethane (8.00 mL), and 4 M hydrogen chloride in dioxane (4.00 mL) were added to the reaction flask and reacted at room temperature for 0.5 h. TLC showed that the reaction of the starting material was complete. The solvent was dried to give 800 mg of a yellow solid crude product, namely compound 133-5.

[0378] Step 6: Synthesis of compound 133-6

[0379] Compound 133-5 (270.00 mg), tetrahydrofuran (10.00 mL), and sodium hydride (154.22 mg) were added to a reaction flask and allowed to react at room temperature for 5 minutes. 7,9-dichloro-10-fluoro-3-methylsulfanyl-2,4,8-triazabicyclo[4.4.0]dec-2,4,6,8,10-penten-5-ol (450.00 mg) was then added and allowed to react at room temperature for 0.5 hours. The reaction was quenched with aqueous ammonium chloride on ice, extracted with EA, dried, and concentrated. Purification by column chromatography (DCM:methanol = 6:1) afforded 410 mg of a yellow solid, compound 133-6.

[0380] Step 7: Synthesis of compound 133-7

[0381] Compound 133-6 (400.00 mg), N,N-dimethylformamide (0.80 mL), and N,N-diisopropylethylamine (0.51 mL, 3.09 mmol) were added to the reaction flask, and N,N,N′,N′-tetramethyl-O-(7-azabenzotriazole-1-yl)urea hexafluorophosphate (586.73 mg) was added under stirring. The reaction was carried out at room temperature for 3 h. Water was slowly added at room temperature to precipitate a solid, which was filtered and the filter cake was washed three times with water. The filter cake was dried to obtain 230 mg of a light yellow solid, namely compound 133-7.

[0382] Step 8: Synthesis of compound 133-8

[0383] To a reaction flask were added compound 133-7 (220.0 mg), 2-[8-ethyl-7-fluoro-3-(methoxymethoxy)naphthalen-1-yl]-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (427.44 mg), [n-butyldi(1-adamantyl)phosphine](2-amino-1,1'-biphenyl-2-yl)palladium(II) methanesulfonate (43.19 mg), potassium phosphate (377.81 mg), 1,4-dioxane (5.00 mL), and water (0.90 mL). After nitrogen replacement, the reaction was carried out at 90°C under nitrogen protection for 2 h. The mixture was diluted with water, extracted with EA, dried, and concentrated. Purification by column chromatography (DCM:EA = 5:1) afforded 300 mg of a yellow solid, compound 133-8.

[0384] Step 9: Synthesis of compound 133-9

[0385] Compound 133-8 (300.0 mg), dichloromethane (6.00 mL), and m-chloroperbenzoic acid (264.03 mg) were added to a reaction flask and reacted at room temperature for 0.3 h. Aqueous sodium bicarbonate solution was added to adjust the pH to 8, and the mixture was extracted with DCM, dried, and concentrated to obtain 280 mg of a crude yellow solid, namely compound 133-9.

[0386] Step 10: Synthesis of compound 133-10

[0387] Compound 133-9 (270.00 mg), [1-(hydroxymethyl)cyclopropyl]methanol (91.82 mg), tetrahydrofuran (6.00 mL), and sodium tert-butoxide (86.39 mg) were added to a reaction flask and reacted at room temperature for 0.2 h. The reaction was quenched by adding aqueous ammonium chloride under an ice bath, extracted with EA, dried, and concentrated. Purification by column chromatography (DCM:EA=1:1) gave 180 mg of a yellow solid, compound 133-10. Step 11: Synthesis of compound 133-11

[0388] Compound 133-10 (180.00 mg), dichloromethane (5.00 mL), and Dess-Martin periodinane (183.95 mg) were added to a reaction flask and reacted at room temperature for 0.5 h. Aqueous sodium bicarbonate solution was added to adjust the pH to 8, and the mixture was extracted with DCM, dried, and concentrated. Purification by column chromatography (DCM:EA = 2:1) afforded 120 mg of a yellow solid, compound 133-11.

[0389] Step 12: Synthesis of compound 133-12

[0390] Compound 133-11 (120.00 mg), intermediate M6 (140.27 mg), 1,2-dichloroethane (4.00 mL), MeOH / methanol (4.00 mL), and 1M zinc chloride in tetrahydrofuran solution (0.19 mL) were added to the reaction flask, and the reaction was carried out at 40°C for 1 h. Sodium cyanoborohydride (121.47 mg) was added and the reaction was continued at 60°C for 3 h. The product was directly purified by silica gel column chromatography (DCM: methanol = 15:1) to obtain 180 mg of a yellow solid, namely compound 133-12.

[0391] Step 13: Synthesis of compound 133

[0392] Compound 133-12 (170.00 mg), dichloromethane (2.00 mL), and trifluoroacetic acid (2.00 mL) were added to a reaction flask and reacted at room temperature for 0.5 h. The mixture was concentrated, dissolved in DCM, and then added to an icy sodium bicarbonate solution adjusted to pH 8. The mixture was extracted with DCM and methanol, dried, and concentrated. Pre-TLC purification (DCM:methanol = 10:1) afforded 88.6 mg of compound 133. LCMS: 1 / 2 [M+2H] + =611.02

[0393] 1 H NMR (500MHz, Methanol-d4) δ8.86(d,J=7.7Hz,1H),8.72(s,1H),8.55(d,J=13.2Hz,1H),8.17(d,J=5.7Hz ,1H),7.62(d,J=7.6Hz,1H),7.42(d,J=5.6Hz,4H),7.23(dd,J=21.4,12.2Hz,3H),5.54–5.32(m,2H),5.0 6(m,2H),4.61–4.36(m,5H),3.97–3.73(m,3H),3.15–2.89(m,3H),2.70–2.41(m,9H),2.08(t,J=58.0Hz, 6H),1.73(m,6H),1.57–1.39(m,6H),1.28(m,1H),1.16(d,J=6.0Hz,3H),0.95–0.73(m,8H),0.56(s,2H).

[0394] Example 134: Synthesis of Compound (2S,4R)-1-((2S)-2-(4-(5-(1-((7-(8-ethyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluoro-4-(3-hydroxy-3-methylazepin-1-yl)pyrido[4,3-d]pyrimidin-2-yl)oxy)methyl)cyclopropyl)methyl)piperidin-4-yl)oxy)pyrazin-2-yl)-1H-1,2,3-triazol-1-yl)-3-methylbutyryl)-4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)ethylpyrrolidine-2-carboxamide

[0395] Step 1: Synthesis of compound 134-1

[0396] 3-Hydroxy-3-methylazepane-1-carboxylic acid tert-butyl ester (1.0 g) was dissolved in DCM (20 mL), and 10 mL of hydrogen chloride in dioxane was added. The mixture was stirred for 30 min. After the reaction was complete, the mixture was concentrated to give compound 134-1 (0.56 g). ESI-MS m / z: 130.1 [M+H] + .

[0397] Step 2: Synthesis of compound 134-2

[0398] 2,4,7-Trichloro-8-fluoropyrido[4,3-d]pyrimidine (1.0 g) was dissolved in DCM (20 mL), cooled to approximately -40°C, and DIPEA (1.38 g) was added. The mixture was stirred at low temperature for 30 minutes, and 134-1 (0.53 g) was slowly added. The reaction was stirred for 1.0 hour. After the reaction was complete, the mixture was diluted with water, extracted with DCM, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (EA:DCM = 1:10) to obtain compound 134-2 (1.0 g). ESI-MS m / z: 345.3 [M+H] + .

[0399] Step 3: Synthesis of compound 134-3

[0400] Compound 134-2 (1.0 g), cesium carbonate (2.1 g), DABCO (0.16 g), and 1,1-cyclopropane dimethanol (0.67 g) were dissolved in DMF (10 mL). The reaction was stirred at room temperature for 1 hour. After the reaction was complete, the mixture was diluted with water, extracted with EA, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (EA:DCM = 1:4) to obtain compound 134-3 (0.8 g). ESI-MS m / z: 411.1 [M+H] + .

[0401] Step 4: Synthesis of compound 134-4

[0402] To a solution of compound 134-3 (0.8 g) in DCM (20 mL) was added DMP (3 g) in portions and stirred at room temperature for 3.0 h. After the reaction was complete, the mixture was quenched with saturated sodium sulfite, diluted with water, and extracted with DCM. The aqueous phase was extracted twice more with DCM. The combined organic phases were dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (PE:EA = 1:1) to afford compound 134-4 (0.67 g). ESI-MS m / z: 409.1 [M+H] + .

[0403] Step 5: Synthesis of compound 134-5

[0404] Compound 134-4 (0.67 g), M4 (0.9 g), potassium carbonate (0.93 g), and Cataxium A Pd G3 (0.21 g) were dissolved in 1,4-dioxane (5 mL) and H2O (1 mL) and reacted at 90°C for 1 h under nitrogen. After the reaction was complete, the reaction solution was diluted with EA and H2O, extracted with EA, washed once with saturated brine, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (EA:DCM = 1:4) to obtain compound 134-5 (0.27 g). ESI-MS m / z: 562.3 [M+H] + .

[0405] Step 6: Synthesis of compound 134

[0406] 134-5 (141.0 mg) and M6 (120 mg) were dissolved in MeOH (5 mL), and a 1 M zinc chloride solution in tetrahydrofuran (0.3 mL) was added. The mixture was reacted at 40°C for 1.5 hours. Sodium cyanoborohydride (45 mg) was added to the reaction solution, and the reaction was continued at 60°C for 8 hours. The reaction was monitored by LCMS to be complete. Saturated ammonium chloride solution (5 mL) was added to the reaction solution, the organic phase was concentrated, DCM (10 mL) was added, and the mixture was washed once with saturated brine. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and subjected to preparative liquid separation to obtain 70.7 mg of compound 134. ESI-MS m / z: 637.8 1 / 2 [M+2H] + .

[0407] Unless otherwise specified, other compounds in the present invention can be prepared by methods similar to those of the above examples.

[0408] Biological experiments

[0409] Biology Experiment 1: Cell Proliferation Experiment

[0410] Table 1

[0411] Cells with different mutations were plated in low-adsorption transparent 96-well plates according to the plating density in Table 1 and cultured overnight in a cell culture incubator. After the cells adhered, the test compounds were added to the 96-well plates at final concentrations of 20,000, 5,000, 1,250, 312.5, 78.13, 19.53, 4.88, 1.22, 0.31, and 0 nM (DMSO final concentration was 0.25%). After 96 hours of culture at 37°C, 50 μL of Cell-titer Glo working solution was added to each well, shaken to mix, and incubated at room temperature for 10 minutes. Luminescence values ​​were read on a multifunctional microplate reader, and the luminescence value data was converted into inhibition percentage. The percentage of cell proliferation inhibition was calculated according to the following formula:

[0412] Inhibition percentage = (1-(measured value-Blank) / (maximum value-Blank))*100%

[0413] ("Maximum value" is from 0.25% DMSO control wells, "Blank" is from blank medium control wells, and "Measured value" is from compound-treated wells).

[0414] GraphPad Prism software was used for curve fitting and IC 50 (nM) values, the results are shown in Table 2.

[0415] Table 2

[0416] “ / ” indicates not determined.

[0417] Biological Experiment 2. Western Blotting Experiment of Compound 115

[0418] Cell plating

[0419] Take AGS cells that are in the logarithmic growth phase and in good growth condition, discard the culture medium, wash once with PBS, and add 0.25% Trypsin to digest the cells;

[0420] Add culture medium to stop digestion, collect cells in a 15 mL centrifuge tube, centrifuge at 1000 rpm for 3 min, discard the culture medium, add 2 mL culture medium to resuspend, count, and dilute with culture medium to 3.33 x 10 5 cells / mL(3x10 5 cells / 0.9mL / well);

[0421] The cells were plated in a 12-well plate at a volume of 0.9 mL per well and cultured overnight at 37°C in the presence of 5% CO2.

[0422] Dosing treatment

[0423] Preparation of compound working solution: aspirate a certain volume of stock solution and dilute it with DMSO to a 1000x intermediate solution. Then, use DMSO to make gradient dilutions to obtain intermediate solutions of various concentrations. Then, pipette 3uL of the intermediate solution of each concentration into 300uL of culture medium and mix thoroughly by pipetting to prepare a 10x working solution.

[0424] Add drugs: pipette 100uL of the corresponding working solution into each well, mix well, and then place the 12-well plate at 37°C and 5% CO2 for 24 hours.

[0425] Protein collection and quantification

[0426] Prepare cell lysis buffer: Take out the prepared 10x RIPA lysis buffer, 10x phosphatase inhibitor, and 25x protease inhibitor from -20℃ in advance, dissolve them at room temperature, and prepare cell lysis buffer according to the following ratio: 760uL ddH2O, 100uL 10x RIPA lysis buffer, 100uL 10x phosphatase inhibitor, and 40uL 25 protease inhibitor per ml, mix well, and set aside;

[0427] Lyse cells: After the drug treatment time is over, carefully aspirate the culture medium in the wells, add 500uL PBS to each well and gently rinse, aspirate the PBS, slowly add 50uL cell lysis buffer from the center of the well, lyse on ice for 10-15 minutes, collect the lysate into a 1.5mL centrifuge tube, centrifuge at 15000rpm at 4℃ for 15 minutes, and aspirate the supernatant into another clean centrifuge tube;

[0428] Protein determination: Take a clean transparent 96-well plate, add 20uL PBS to each well, then add 5uL of the collected protein supernatant, test in a single well, then premix BCA solution A and B solution at a ratio of 50:1, add 200uL of the mixed BCA colorimetric solution to each well, wrap the 96-well plate with tin foil, incubate at 37°C for 30 minutes, and use an Evisin instrument to measure the OD value (562nM), substitute it into the standard curve, and calculate the protein concentration.

[0429] Perform WB experiments

[0430] 1. Adjust protein concentration: According to the calculation formula, use PBS to adjust the protein loading amount per well to 30μg, and the final volume is 10μL. Add 2μL 6x SDS loading buffer to each sample, cook at 100℃ for 5min, then centrifuge and cool to room temperature;

[0431] 2. Assemble the electrophoresis tank, add electrophoresis buffer, load 12uL of sample into each well, and add 2.5uL and 1.5uL of marker to the left and right of the sample respectively. Fill the marker well and the remaining wells with 1x loading buffer to 12uL. Run electrophoresis at 90V for about 30min. After the sample is compressed into a single band in the gel, increase the voltage to 120V and run the gel until the blue loading buffer band reaches the bottom of the gel.

[0432] 3. Prepare 2-3 L of transfer buffer in advance at a ratio of 7:2:1 (ddH2O:methanol:10x transfer buffer) and cool at 4°C.

[0433] 4. After running the gel, carefully disassemble the gel sheet, remove the gel, cut off the useless part, place it on the transfer clamp padded with filter paper, and gently cover it with the cut PVDF membrane (PVDF membrane needs to be activated by soaking in methanol in advance and soaked in transfer buffer). Use a roller to carefully roll back and forth to remove bubbles between the gel and membrane, then cover it with filter paper, carefully tighten the transfer clamp, insert it into the transfer tank, add transfer buffer, and place the transfer tank in a slightly larger container filled with crushed ice. Transfer the membrane at 400mA for 120-150min (depending on the molecular weight of the protein).

[0434] 5. Prepare 5% BSA blocking solution: weigh 2.5 g BSA, dissolve in TBS-T, and dilute to 50 mL;

[0435] 6. After transfer, carefully remove the PVDF membrane and soak it in 5% BSA solution. Block the membrane on a shaker at room temperature for 1 hour. 7. Prepare the primary antibody: Dilute 5% BSA to 2% to dilute the antibody. Dilute the primary antibody according to the recommended ratio in the antibody manual.

[0436] 8. Transfer the blocked PVDF membrane to the primary antibody solution and incubate at 4°C overnight;

[0437] Incubation with secondary antibody

[0438] 1. Remove the PVDF membrane from the primary antibody solution and wash it three times with TBS-T on a shaker, each time for 10 minutes. The used primary antibody can be stored at -20°C and reused;

[0439] 2. According to the instructions for the primary antibody, dilute the corresponding rabbit or mouse antibody with 2% BSA at a dilution ratio of 4000:1. Place the PVDF membrane in the secondary antibody solution and incubate at room temperature for 1-2 hours.

[0440] 3. After the secondary antibody incubation, wash three times with TBS-T, each time for 10 minutes;

[0441] Luminescence Steps

[0442] 1. Turn on the luminometer and select BioSense mode;

[0443] 2. Mix luminescent liquid A and liquid B in equal proportions in a container. After wiping the PVDF membrane dry with dust-free paper, place it in the container. Use a pipette to draw up the luminescent liquid and repeatedly pour it on the PVDF membrane. After the PVDF membrane is fully soaked with the luminescent liquid, place the membrane in the instrument for exposure.

[0444] The other two cell lines, Mia-Paca2 and SW620, were obtained by referring to the above test method, and the results are shown in Figure 1.

[0445] Biological Experiment 3. PK Study of Compound 115 in ICR Mice

[0446] Experimental methods and results:

[0447] The compound was mixed with the solvent 10% DMSO / 5% Solutol / 85% saline, vortexed, and sonicated to prepare a 0.6 mg / ml clear solution. Compound 115 was administered intravenously to 6- to 7-week-old female ICR mice without fasting. Whole blood was collected over time for drug concentration analysis using LC-MS / MS, and pharmacokinetic parameters were calculated using Phoenix WinNolin software (Pharsight, USA). The results are shown in Table 3.

[0448] Table 3. PK data of intravenous administration of compound 115 in mice

[0449] Biological Experiment 4. PK Study of Compound 115 in SD Rats

[0450] Experimental methods and results:

[0451] The compound was mixed with the solvent 10% DMSO / 5% Solutol / 85% saline, vortexed, and sonicated to prepare a 1.5 mg / ml clear solution. Six- to seven-week-old male Sprague-Dawley rats were fasted for 8 hours and then intravenously administered compound 115. Whole blood was collected over time for drug concentration analysis using LC-MS / MS, and pharmacokinetic parameters were calculated using Phoenix WinNolin software (Pharsight, USA). The results are shown in Table 4.

[0452] Table 4. PK data of intravenous administration of compound 115 in rats

[0453] Biological Experiment 5. PK Study of Compound 115 in Beagle Dogs

[0454] Experimental methods and results:

[0455] The compound was mixed with the solvent 10% DMSO / 5% Solutol / 85% saline, vortexed, and sonicated to prepare a 1.5 mg / ml clear intravenous solution. Compound 115 was administered intravenously to approximately 10 kg beagle dogs (half male and half female) without fasting. Whole blood samples were collected intravenously at 5 min, 15 min, 30 min, 1 h, 2 h, 4 h, 6 h, 8 h, 10 h, 24 h, 48 h, and 72 h. Drug concentrations were analyzed by LC-MS / MS, and pharmacokinetic parameters were calculated using Phoenix WinNolin software (Pharsight, USA). The results are shown in Table 5. The dosing schedule and results are shown in Table 5.

[0456] Table 5. PK data of intravenous compound 115 in beagle dogs

[0457] Although the present invention has been fully described through its embodiments, it is noteworthy that various changes and modifications are obvious to those skilled in the art. Such changes and modifications should be included within the scope of the appended claims of the present invention.

Claims

1. A compound having the structure of FLM, or its tautomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, characterized in that: The F is a KRAS protein binding fragment represented by general formula (I); The K is selected from a 6-12 membered heterocyclic group containing N; the K may be optionally further replaced by one or more R a replaced by; The R 14 is selected from halogen; R6 is selected from H, hydroxyl, amino, cyano, halogen, C 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Haloalkoxy or C 1-6 Alkoxy; preferably H; Alternatively, the R6 and K and the atoms to which they are connected together form a 3-14 membered heterocyclic group; wherein the 3-14 membered heterocyclic group may be further optionally replaced by one or more R a replaced by; The L is a connecting unit connecting F and M; the L is selected from in, The G ring and the D ring are each independently selected from C 6-14 Aryl, 5-14 membered heteroaryl, C 3-14 Cycloalkyl or 3-14 membered heterocyclic group; wherein the C 6-14 Aryl, 5-14 membered heteroaryl, C 3-14 The cycloalkyl or 3-14 membered heterocyclic group is optionally further substituted with one or more R a replace; The A rings are each independently selected from 3-7 membered N-containing heterocyclic groups; wherein the 3-7 membered N-containing heterocyclic groups are optionally further substituted by one or more R a Preferably, the A ring is independently selected from a 3-6 membered N-containing heterocyclic group; the 3-6 membered N-containing heterocyclic group is optionally further substituted by one or more R a replace; The E's are each independently absent or selected from -O-, -NH-, or -NCH3-; The R7 and R8 are each independently selected from H, halogen, hydroxyl, amino, oxo, cyano, halogen, C 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Haloalkoxy or C 1-6 Alkoxy; Alternatively, R7, R8 and the atoms to which they are connected together form C 3-14 Cycloalkyl or 3-14 membered heterocyclic group; wherein the C 3-14 The cycloalkyl or 3-14 membered heterocyclic group is optionally further substituted with one or more R a replace; The n1, n2, n3, and n4 are each independently selected from an integer of 0 to 10, preferably 0, 1, 2, 3 or 4; The M is selected from the VHL binding fragment represented by the general formula (II); Wherein, X1 is selected from C, CH or N; Said X2 is selected from C, CH or N; Said X3 is selected from N, NH, O or S; The R1 is selected from H, C 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Alkoxy, C 1-6 Haloalkoxy, halogen or cyano; preferably methyl, ethyl, F or cyano; The R2 is selected from H, C 1-6 Alkyl or C 1-6 Haloalkyl; preferably H or methyl; The R3 is selected from H, C 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Hydroxyalkyl, -(CH2) n N(R9)2, -(CH2) n -CO-OR9 or -(CH2) n -CO-N(R9)2, wherein R9 is independently selected from H, C 1-6 Alkyl, C 1-6 Haloalkyl or C 1-6 Alkoxy; preferably, the -(CH2) n -CO-N(R9)2 is -(CH2) n -CO-NH-(CH2) n -CH3; Said n is selected from 0, 1, 2 or 3; The R4 is selected from or -NH-; R5 is selected from H, C 1-6 Alkyl or C 1-6 Haloalkyl; The R a are independently selected from H, hydroxy, amino, oxo, cyano, halogen, C 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Alkoxy, -C 0-3 Alkylene-OR b 、-OC(=O)C 1-6 Alkyl, -C 0-3 Alkylene-SR b , -C 0-3 Alkylene-N(R b 2. -C 0-3 Alkylene-S(=O)R b , -C 0-3 Alkylene-S(=O)2R b , -C 0-3 Alkylene-SR b , -C 0-3 Alkylene-S(R b )5. -C 0-3 Alkylene-C(=O)R b , -C 0-3 Alkylene-C(=O)OR b , -C 0-3 Alkylene-C(=O)N(R b 2. C 2-6 Alkenyl, C 2-6 Alkynyl, -C 0-3 Alkylene-C 3-14 Cycloalkyl, -C 0-3 Alkylene-(3-14 membered heterocyclic group), -C 0-3 Alkylene-C 6-18 Aryl or -C 0-3 Alkylene-(5-18 membered heteroaryl); wherein the hydroxyl, amino, C 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Alkoxy, C 0-3 Alkylene, C 2-6 Alkenyl, C 2-6 Alkynyl, -C 0-3 Alkylene-C 3-14 Cycloalkyl, -C 0-3 Alkylene-(3-14 membered heterocyclic group), -C 0-3 Alkylene-C 6-18 Aryl or -C 0-3 Alkylene-(5-18 membered heteroaryl) is optionally further substituted with one or more R b replace; Each R b are independently H, halogen, hydroxy, amino, cyano, C 1-6 Alkyl, C 3-6 Cycloalkyl, C 1-6 Haloalkyl, C 3-14 Cycloalkyl, 3-14 membered heterocyclic group, C 6-18 Aryl or 5-18 membered heteroaryl; wherein the hydroxyl, amino, C 1-6 alkyl Base, C 3-6 Cycloalkyl, C 1-6 Haloalkyl, C 3-14 Cycloalkyl, 3-14 membered heterocyclic group, C 6-18 The aryl or 5-18 membered heteroaryl is optionally further substituted with one or more halogen, C 1-6 Alkyl or C 1-6 or two R b Together with its co-attached atoms, it forms C 3-14 Cycloalkyl, 3-14 membered heterocyclic group, C 6-18 Aryl or 5-18 membered heteroaryl, the C 3-14 Cycloalkyl, 3-14 membered heterocyclic group, C 6-18 The aryl or 5-18 membered heteroaryl is optionally further substituted with one or more halogen, amino, hydroxyl, cyano, C 1-6 Alkyl, C 3-6 Cycloalkyl or C 1-6 Haloalkyl substitution.

2. The compound according to claim 1, or its tautomer, racemate, enantiomer, diastereomer, or mixture thereof or a pharmaceutically acceptable salt thereof, characterized in that: The F is a KRAS protein binding fragment represented by general formula (I); The K is selected from a 6-12 membered heterocyclic group containing N; the K may be optionally further replaced by one or more R a replaced by; The R 14 is selected from halogen; R6 is selected from H, hydroxyl, amino, cyano, halogen, C 1-6 Alkyl, C 1-6 Haloalkyl or C 1-6 Alkoxy; preferably H; Alternatively, the R6 and K and the atoms to which they are connected together form a 3-14 membered heterocyclic group; wherein the 3-14 membered heterocyclic group may be further optionally replaced by one or more R a replaced by; The L is a connecting unit connecting F and M; the L is selected from in, The G ring and the D ring are each independently selected from C 6-14 Aryl, 5-14 membered heteroaryl, C 3-14 Cycloalkyl or 3-14 membered heterocyclic group; wherein the C 6-14 Aryl, 5-14 membered heteroaryl, C 3-14 The cycloalkyl or 3-14 membered heterocyclic group is optionally further substituted with one or more R a replace; The A rings are each independently selected from 3-7 membered N-containing heterocyclic groups; wherein the 3-7 membered N-containing heterocyclic groups are optionally further substituted by one or more R a Preferably, the A ring is independently selected from a 3-6 membered N-containing heterocyclic group; wherein the 3-6 membered N-containing heterocyclic group is optionally further replaced by one or more R a replace; The E's are each independently absent or selected from -O-, -NH-, or -NCH3-; The R7 and R8 are each independently selected from H, halogen, hydroxyl, amino, oxo, cyano, halogen, C 1-6 Alkyl, C 1-6 Haloalkyl or C 1-6 Alkoxy; Alternatively, R7, R8 and the atoms to which they are connected together form C 3-14 Cycloalkyl or 3-14 membered heterocyclic group; wherein the C 3-14 The cycloalkyl or 3-14 membered heterocyclic group is optionally further substituted with one or more R a replace; The n1, n2, n3, and n4 are each independently selected from an integer of 0 to 10, preferably 0, 1, 2, 3 or 4; The M is selected from the VHL binding fragment represented by the general formula (II); Wherein, X1 is selected from C, CH or N; Said X2 is selected from C, CH or N; Said X3 is selected from N, NH, O or S; The R1 is selected from H, C 1-6 Alkyl, halogen or cyano, preferably methyl, ethyl, F or cyano; R2 is selected from H or C 1-6 Alkyl, preferably H or methyl; The R3 is selected from H, C 1-6 Alkyl, C 1-6 Hydroxyalkyl, -(CH2) n N(R9)2, -(CH2) n -CO-OR9 or -(CH2) n -CO-N(R9)2, wherein R9 is independently selected from H, C 1-6 Alkyl, C 1-6 Haloalkyl or C 1-6 Alkoxy; preferably, the -(CH2) n -CO-N(R9)2 is -(CH2) n -CO-NH-(CH2) n -CH3; Said n is selected from 0, 1, 2 or 3; The R4 is selected from or -NH-; R5 is selected from H, C 1-6 Alkyl or C 1-6 Haloalkyl; The R a are independently selected from H, hydroxy, amino, oxo, cyano, halogen, C 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Alkoxy, -C 0-3 Alkylene-OR b 、-OC(=O)C 1-6 Alkyl, -C 0-3 Alkylene-SR b , -C 0-3 Alkylene-N(R b 2. -C 0-3 Alkylene-S(=O)R b , -C 0-3 Alkylene-S(=O)2R b , -C 0-3 Alkylene-SR b , -C 0-3 Alkylene-S(R b )5. -C 0-3 Alkylene-C(=O)R b , -C 0-3 Alkylene-C(=O)OR b , -C 0-3 Alkylene-C(=O)N(R b 2. C 2-6 Alkenyl, C 2-6 Alkynyl, -C 0-3 Alkylene-C 3-14 Cycloalkyl, -C 0-3 Alkylene-(3-14 membered heterocyclic group), -C 0-3 Alkylene-C 6-18 Aryl or -C 0-3 Alkylene -(5-18 membered heteroaryl); wherein the C 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Alkoxy, C 2-6 Alkenyl, C 2-6 Alkynyl, -C 0-3 Alkylene-C 3-14 Cycloalkyl, -C 0-3 Alkylene-(3-14 membered heterocyclic group), -C 0-3 Alkylene-C 6-18 Aryl or -C 0-3 Alkylene-(5-18 membered heteroaryl) is optionally further substituted with one or more R b replace; Each R b are independently H, halogen, hydroxyl, cyano, C 1-6 Alkyl, C 3-6 Cycloalkyl, C 1-6 Haloalkyl, C 3-14 Cycloalkyl, 3-14 membered heterocyclic group, C 6-18 Aryl or 5-18 membered heteroaryl; wherein the C 1-6 Alkyl, C 3-6 Cycloalkyl, C 1-6 Haloalkyl, C 3-14 Cycloalkyl, 3-14 membered heterocyclic group, C 6-18 The aryl or 5-18 membered heteroaryl is optionally further substituted with one or more halogen, C 1-6 Alkyl or C 1-6 or two R b Together with its co-attached atoms, it forms C 3-14 Cycloalkyl, 3-14 membered heterocyclic group, C 6-18 Aryl or 5-18 membered heteroaryl, the C 3-14 Cycloalkyl, 3-14 membered heterocyclic group, C 6-18 The aryl or 5-18 membered heteroaryl is optionally further substituted with one or more halogen, amino, hydroxyl, cyano, C 1-6 Alkyl, C 3-6 Cycloalkyl or C 1-6 Haloalkyl substitution.

3. The compound according to claim 1 or 2, or its tautomer, racemate, enantiomer, diastereomer, or mixture thereof or a pharmaceutically acceptable salt thereof, characterized in that: The K is selected from 4. A compound according to any one of claims 1 to 3, or a tautomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, characterized in that: The K is selected from 5. The compound according to any one of claims 1 to 4, or its tautomer, racemate, enantiomer, diastereomer, or mixture thereof or a pharmaceutically acceptable salt thereof, characterized in that: The K is selected from 6. The compound according to any one of claims 1 to 5, or its tautomer, racemate, enantiomer, diastereomer, or mixture thereof or a pharmaceutically acceptable salt thereof, characterized in that: The F is selected from 7. A compound according to any one of claims 1 to 6, or a tautomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, characterized in that: The R4 is 8. A compound according to any one of claims 1 to 6, or a tautomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, characterized in that: The R4 is 9. The compound according to any one of claims 1 to 6, or its tautomer, racemate, enantiomer, diastereomer, or mixture thereof or a pharmaceutically acceptable salt thereof, characterized in that: The R4 is -NH-.

10. The compound according to any one of claims 1 to 9, or its tautomer, racemate, enantiomer, diastereomer, or mixture thereof or a pharmaceutically acceptable salt thereof, characterized in that: The G ring or D ring in L is independently selected from C 6-14 Aryl, 5-14 membered heteroaryl, C 3-14 Cycloalkyl or 3-14 membered heterocyclic group; wherein the C 6-14 Aryl, 5-14 membered heteroaryl, C 3-14 The cycloalkyl or 3-14 membered heterocyclic group is optionally further substituted with one or more halogen or C 1-3 Alkyl substitution; The C 6-14 The aryl group is preferably The 5-14 membered heteroaryl group is preferably selected from The 3-14 membered heterocyclic group is preferably selected from The C 3-14 The cycloalkyl group is preferably selected from 11. The compound according to any one of claims 1 to 10, or its tautomer, racemate, enantiomer, diastereomer, or mixture thereof or a pharmaceutically acceptable salt thereof, characterized in that: The D rings are each independently selected from 5-6 membered nitrogen-containing heteroaryl groups; wherein the 5-6 membered nitrogen-containing heteroaryl groups are optionally further substituted with one or more R a replace; The G rings are each independently selected from C 6-14 Aryl, 5-14 membered heteroaryl, C 3-14 Cycloalkyl or 3-14 membered heterocyclic group; wherein the C 6-14 Aryl, 5-14 membered heteroaryl, C 3-14 The cycloalkyl or 3-14 membered heterocyclic group is optionally further substituted with one or more R a replace; The A rings are each independently selected from 3-7 membered N-containing heterocyclic groups; wherein the 3-7 membered N-containing heterocyclic groups are optionally further substituted by one or more R a replace; The E's are each independently absent or selected from -O-, -NH-, or -NCH3-; The R7 and R8 are each independently selected from H, halogen, hydroxyl, amino, oxo, cyano, halogen, C 1-6 Alkyl, C 1-6 Haloalkyl or C 1-6 Alkoxy; Alternatively, the R7, R8 and the atoms to which they are connected together form a C 3-14 Cycloalkyl or 3-14 membered heterocyclic group; wherein the C 3-14 The cycloalkyl or 3-14 membered heterocyclic group is optionally further substituted with one or more R a replace; The n1, n2, n3, and n4 are each independently selected from an integer of 0-10, preferably 0, 1, 2, 3 or 4.

12. A compound according to any one of claims 1 to 11, or a tautomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, characterized in that: The A rings are each independently selected from 3-7 membered N-containing heterocyclic groups; wherein the 3-7 membered N-containing heterocyclic groups are optionally further substituted with one or more halogens or C 1-3 Preferably, the A rings are each independently selected from 3-6 membered N-containing heterocyclic groups; wherein the 3-6 membered N-containing heterocyclic groups are optionally further substituted with one or more halogens or C 1-3 Alkyl substitution.

13. The compound according to claim 12, or its tautomer, racemate, enantiomer, diastereomer, or mixture thereof or a pharmaceutically acceptable salt thereof, characterized in that: The 3-7 membered N-containing heterocyclic group is 14. A compound according to any one of claims 1 to 13, or a tautomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, characterized in that: The E is -O-.

15. The compound according to any one of claims 1 to 14, or its tautomer, racemate, enantiomer, diastereomer, or mixture thereof or a pharmaceutically acceptable salt thereof, characterized in that: The E is -NH-.

16. A compound according to any one of claims 1 to 15, or a tautomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, characterized in that: The E is -NCH3-.

17. A compound according to any one of claims 1 to 16, or a tautomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, characterized in that: The D ring is independently selected from a 5-6 membered nitrogen-containing heteroaryl group, and the 5-6 membered heteroaryl group is selected from wherein the 5-6 membered nitrogen-containing heteroaryl group is optionally further substituted by one or more R a Substitution; said R a is selected from H, hydroxy, amino, oxo, cyano, halogen, C 1-6 Alkyl, C 1-6 Haloalkyl or C 1-6 Alkoxy.

18. A compound according to any one of claims 1 to 17, or a tautomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, characterized in that: The G ring is independently selected from C 6-14 Aryl, 5-14 membered heteroaryl, C 3-14 Cycloalkyl or 3-14 membered heterocyclic group; wherein the C 6-14 Aryl, 5-14 membered heteroaryl, C 3-14 The cycloalkyl or 3-14 membered heterocyclic group is optionally further substituted with one or more R a replace; The C 6-14 The aryl group is preferably The 5-14 membered heteroaryl group is preferably selected from The 3-14 membered heterocyclic group is preferably selected from The C 3-14 The cycloalkyl group is preferably selected from The R a is selected from H, hydroxy, amino, oxo, cyano, halogen, C 1-6 Alkyl, C 1-6 Haloalkyl or C 1-6 Alkoxy.

19. A compound according to any one of claims 1 to 18, or a tautomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, characterized in that: The G ring is independently selected from a 3-14 membered heterocyclic group, and the 3-14 membered heterocyclic group is optionally further substituted by one or more R a replace; The 3-14 membered heterocyclic group is preferably selected from The R a is selected from H, hydroxy, amino, oxo, cyano, halogen, C 1-6 Alkyl, C 1-6 Haloalkyl or C 1-6 Alkoxy.

20. The compound according to any one of claims 1 to 19, or its tautomer, racemate, enantiomer, diastereomer, or mixture thereof or a pharmaceutically acceptable salt thereof, characterized in that: The L is selected from 21. A compound according to any one of claims 1 to 19, or a tautomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, characterized in that: The L is selected from 22. A compound according to any one of claims 1 to 21, or a tautomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, characterized in that: The R3 is selected from H, methyl, hydroxymethyl, -CH2-CO-O-CH3, -CH2-N(CH3)2 or -CH2-CO-NH-CH3.

23. A compound according to any one of claims 1 to 22, or a tautomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, characterized in that: The M is selected from 24. The compound according to claim 1, or its tautomer, racemate, enantiomer, diastereomer, or mixture thereof or a pharmaceutically acceptable salt thereof, characterized in that: The compound of the FLM structure is selected from the following compounds:

25. A pharmaceutical composition comprising a compound according to any one of claims 1 to 24, or a tautomer, racemate, enantiomer, diastereomer, or a mixture thereof, or a pharmaceutically acceptable salt thereof.

26. Use of a compound according to any one of claims 1 to 24, or a tautomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 25 in the preparation of a medicament for regulating KRAS protein ubiquitination and degradation in a subject.

27. Use of a compound according to any one of claims 1 to 24, or a tautomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 25 in the preparation of a medicament for treating and / or preventing a disorder mediated by or dependent on the KRAS protein, wherein the disease mediated by the KRAS protein is preferably a tumor.

28. The use according to claim 27, characterized in that The disease is selected from breast cancer, multiple myeloma, bladder cancer, endometrial cancer, gastric cancer, cervical cancer, rhabdomyosarcoma, non-small cell lung cancer, small cell lung cancer, pleomorphic lung cancer, ovarian cancer, esophageal cancer, melanoma, colorectal cancer, hepatoma, head and neck tumors, hepatobiliary carcinoma, myelodysplastic syndrome, malignant glioma, prostate cancer, thyroid cancer, Schwann cell tumor, squamous cell carcinoma of the lung, lichenoid keratosis, synovial sarcoma, skin cancer, pancreatic cancer, testicular cancer or liposarcoma.

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