Heterocyclic compound as kras inhibitor, preparation therefor and therapeutic use thereof

Abstract

The present invention relates to a compound of formula (I) or a pharmaceutically acceptable salt, prodrug, tautomer, stereoisomer or solvate thereof, which compound can be used in the treatment of cancers and inflammation in mammals. Further provided are a method for preparing the compound of formula (I) and a pharmaceutical composition containing the compound.

Description

Heterocyclic compounds as KRAS inhibitors, their preparation and therapeutic uses Technical Field

[0001] This invention relates to certain novel heterocyclic compounds or pharmaceutically acceptable salts thereof, said compounds being used to treat or prevent cancers associated with H-ras, K-ras, or N-ras inhibition. The invention also relates to pharmaceutical compositions comprising said compounds or pharmaceutically acceptable salts thereof, intermediates for preparing said compounds, and methods for treating cancers associated with H-ras, K-ras, or N-ras inhibition using said compounds or pharmaceutically acceptable salts thereof. Background Technology

[0002] In 1982, Weinberg and Barbacid first isolated a transforming gene from a human bladder cancer cell line that could induce malignant transformation in NIH 3T3 cells, while DNA extracted from normal human tissue did not have this effect. Subsequently, Santos and Parada discovered that the aforementioned transforming gene was not a novel gene, but a human homologue of the Harvey murine sarcoma virus ras gene, named H2ras. In the same year, Krontiris discovered a homologue of the Kirsten murine sarcoma virus gene in human lung cancer cells, called K-ras. Another similar gene, called N2ras, was discovered when NIH 3T3 cells were infected with human neuroblastoma DNA; this gene is unrelated to the virus.

[0003] The ras gene is highly conserved throughout evolution and is widely found in various eukaryotes such as mammals, fruit flies, fungi, nematodes, and yeast, suggesting an important physiological function. The mammalian ras gene family has three members: H-ras, K-ras, and N-ras. K-ras has two variants, A and B, in its fourth exon. All ras genes have similar structures, consisting of four exons distributed across approximately 30 kb of DNA. Their encoded product is a protein with a relative molecular mass of 21,000, hence the name P21 protein. It has been shown that H-ras is located on the short arm of human chromosome 11 (11p15.1–p15.3), K-ras on the short arm of chromosome 12 (12p1.1–pter), and N-ras on the short arm of chromosome 1 (1p22–p32). Except for variations in the fourth exon of K-ras, the sequence encoding P21 in each ras gene is evenly distributed across the four exons. However, the sequences and sizes of the introns vary considerably, resulting in significant differences in the overall gene length. For example, human K-ras is 35 kb long, while N-ras is 3 kb long. Because it has two fourth exons, K-ras can be spliced ​​in two ways, but the mRNA encoding K-ras-B has a higher content. Except for K-ras-B, which contains 188 amino acids, the other two Ras proteins each contain 189 amino acids.

[0004] Ras(P21) protein is located on the inner side of the cell membrane and plays an important role in transmitting signals for cell growth and differentiation. It belongs to the guanosine triphosphate (GTP)-binding protein family (a coupling factor in cell signaling), regulating signal transmission through the interconversion of GTP and guanosine diphosphate (GDP). P21 has a strong affinity for both GTP and GDP and exhibits relatively weak GTPase activity. Under normal circumstances, P21 and GDP are in an inactive state. When extracellular growth and differentiation factors transmit signals to P21 on the inner side of the cell membrane, the binding activity of P21 to GTP is enhanced, activating the binding and opening the signaling system. Because P21 possesses GTPase activity, it hydrolyzes GTP to GDP. Upon binding to GDP, P21 becomes inactive, and the signaling system shuts down. While P21's GTPase activity is normally very weak, its hydrolysis rate can increase by 10,000 times when it binds to GTPase activator protein (GAP), thus inactivating P21. After P21 binds to GDP, it can activate guanylate-releasing protein (GNRP). GNRP causes P21 to release GDP and bind GTP. Therefore, through the interconversion of GTP and GDP, the activation and deactivation of the signaling system by P21 can be regulated in a controlled manner, thus completing the process of transmitting growth and differentiation signals into the cell.

[0005] More than one-fifth of cancer patients have Ras gene mutations. These mutations mostly occur on G12, G13 and Q61 residues. The mutations cause GAP protein to fail and Ras signaling to remain in an activated state.

[0006] The RAS gene family is the most commonly mutated gene family in human cancers. RAS mutations are present in 90% of pancreatic cancers, 45% of colon cancers, and 35% of lung cancers. Of the three Ras genes, Kirsten-RAS (KRAS) is the most frequently mutated subtype, accounting for up to 86% of cases, while the other two subtypes, neuroblastoma-RAS (NRAS) and Harvey-RAS (HRAS), have lower mutation rates (11% and 3%, respectively). The KRAS protein has several mutations. The KRAS-G12C mutation is dominant in NSCLC (mutated KRAS-G12C accounts for approximately 45-50%). On the other hand, the KRAS-G12D mutation (where glycine is mutated to aspartic acid at codon-12) is very important in pancreatic cancer (61%), colon cancer (42%), and NSCLC (22%).

[0007] Research on drugs for treating KRAS-G12C mutations has yielded exciting results, with AMG510 and MRTX849 offering hope to cancer patients in clinical trials. However, there are currently no targeted drugs for KRAS G12D mutations, creating a significant clinical need. Therefore, it is necessary to develop new KRAS-G12D inhibitors and other KRAS mutation inhibitors.

[0008] This invention designs and synthesizes a series of chemical molecules with strong bioactivity in inhibiting RAS, and provides a method for treating related cancers by inhibiting H-RAS, K-RAS or N-RAS. Summary of the Invention

[0009] This invention provides compounds having inhibitory activity against KRas mutant proteins (e.g., G12C, G12D, G12V, G12A, G12R, G12S, G13D, and Q61H mutant proteins) and capable of modulating G12D mutant KRAS, HRAS, and / or NRAS proteins, including their stereoisomers, pharmaceutically acceptable salts, tautomers, and prodrugs. Methods for treating various diseases or conditions (such as cancer) using these compounds are also provided.

[0010] In one aspect of the invention, a compound having formula (I) or a pharmaceutically acceptable salt, solvate, tautomer, stereoisomer, or prodrug thereof is provided, wherein the compound of formula (I) is:

[0011] in:

[0012] Y is It is a 4- to 12-element saturated or partially saturated single ring, bridged ring, or helical ring, wherein the saturated or partially saturated single ring, bridged ring, or helical ring is optionally composed of one or more R 1 Replace, X 1 Selected from N and CR 4 ;X 2 Selected from NR 4 , O, or C(R) 4 )2,

[0013] R 1 Selected from hydrogen, halogens, optional halogenated elements, hydroxyl groups, C 1-6 Alkoxy, deuterated C 1-6 Alkoxy, -NR 1a R 1b C replaced by deuterium 1- 6-alkyl, CN, -OR 1a -SR 1a -NR 1a R 1b -S(O)R 1a -S(O)2R 1a -CH2OR 1a -C(O)OR 1a -NR 1a C(O)R 1b -C(O)NR 1a R 1b -S(O)2N(R) 1a R 1b )2 and 5- to 6-membered heteroaryl, wherein R 1a and R 1b Each independently is hydrogen, C 1-6 Alkyl, hydroxyl C 1-6 Alkyl, halogenated or deuterated C 1-6 Alkyl and C 1-6 Alkoxy C 1-6 Alkyl-, C 3-6 cycloalkyl;

[0014] R 4 Selected from hydrogen, halogens, and C atoms optionally substituted with halogens or hydroxyl groups. 1-6 Alkyl, CN, -OR 4a -SR 4a -S(O)R 4a -S(O)2R 4a -C(O)R 4a -C(O)OR 4a -NR 4a C(O)R 4b -C(O)NR 4aR 4b and -S(O)2N(R 4a R 4b )2, where R 4a and R 4b Each is independently hydrogen, a 4-6 membered oxecyclic alkyl group optionally substituted with methyl, dimethyl or isopropyl, C 1-6 Alkyl, hydroxyl C 1-6 Alkyl, Halogenated C 1- 6-alkyl and C 1-6 Alkoxy C 1-6 alkyl-;

[0015] L represents a single bond, -O-, -S-, or -NR. La -、-O-(CR La R Lb ) t -、-S-(CR La R Lb ) t -、-NR c -(CR La R Lb ) t -、-(CR La R Lb ) t -O-、-(CR La R Lb ) t -S-、-(CR La R Lb ) t -NR Lc -, -C(O)-, -SO2-, -SO-, -C(O)-O-, -OC(O)-, -C(O)-NR Lc -or-N Lc C(O)-, where R La R Lb and R Lc Each is independently selected from hydrogen, deuterium, and C. 1-6 Alkyl group, or R group attached to the same carbon atom La and R Lb Together with the attached carbon atom, it forms a C3-C6 cycloalkyl group, wherein t is an integer from 1 to 6;

[0016] R 2 C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 3-8 Cycloalkyl, aryl, heteroaryl, or heterocyclic, wherein the C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C3-8 Cycloalkyl, aryl, heteroaryl, and heterocyclic groups are each independently unsubstituted or converted by deuterium, halogen, cyano, or C. 1-6 Alkyl, Halogenated C 1-6 Alkyl, C 1-6 Alkylene, Halogenated C 1-6 Alkylene, hydroxyl C 1-6 Alkyl, C 1-6 Alkoxy C 1-6 Alkyl, oxo, -OR 2a -C(O)R 2a 、-(CR 2a R 2b ) m -OC(O)NR 2c R 2d -CO2R 2a -CONR 2c R 2d -NR 2c R 2d C 3-8 cycloalkyl, C 3-8 cycloalkyl C 1-6 One or more substitutions of alkyl, aryl, heteroaryl, and heterocyclic groups, wherein R 2a R 2b R 2c and R 2d Each is independently hydrogen, C 1-6 Alkyl, hydroxyl C 1-6 Alkyl, Halogenated C 1-6 Alkyl and C 1-6 Alkoxy C 1-6 Alkyl group, or R atom attached to the same nitrogen atom 2c and R 2d Together with the attached nitrogen atom, it forms a 4- to 6-membered heterocycle, the heterocycle containing 0, 1 or 2 additional heteroatoms selected from nitrogen, oxygen or sulfur as ring members, and wherein m is an integer from 1 to 6;

[0017] R 3 It is an aryl or heteroaryl group, wherein the aryl or heteroaryl group is optionally composed of one or more R groups. 8 Replace; each R 8 Independently selected C groups are chosen from halogens, cyano groups, oxo groups, and C groups optionally substituted with halogens, cyano groups, hydroxyl groups, and deuterated groups. 1-6 Alkyl, C2-C6 alkenyl group optionally substituted with hydroxyl or deuterium, C2-C6 ynyl group optionally substituted with hydroxyl or deuterium, -OR 8a -SR 8a -S(O)2R 8a -P(=O)R 8a R 8b -NR8a R 8b -C(O)NR 8a R 8b Optional halogenated or C 1-6 Alkyl-substituted C3-C6 cycloalkyl, C3-C8 cycloalkyl, heterocyclic, heteroaryl, and aryl groups, wherein R 8a and R 8b Each independent hydrogen atom, and C atoms that can be arbitrarily substituted with halogens. 1-6 Alkyl and C 1-6 Alkoxy-C 1-6 alkyl-;

[0018] Q 1 Q 2 and Q 3 Each is independently N or CR 6 M 1 and M 2 Each is independently N or CR 7 The condition is Q 1 and M 1 At least one of them is N;

[0019] Where R 6 and R 7 Each is independently hydrogen, halogen, hydroxyl, cyano, nitro, C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 3- 8-Cycloalkyl, aryl, heteroaryl or heterocyclic, -OR 6a -C(O)R 6a -CO2R 6a -CONR 6a R 6b or -NR 6a R 6b Wherein C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 3-8 Cycloalkyl, aryl, heteroaryl, and heterocyclic groups are each independently converted by oxidizing, halogenating, hydroxyling, or C-oxidizing. 1-4 Alkoxy, C 1-4 Alkyl, C 3-6 Cycloalkyl, nitro, cyano and -NR d R e One or more substitutions in, where R 6a R 6b R 6c and R 6d Each is independently hydrogen, C 3-6 Alkyl, C 1-6 Alkyl, hydroxyl C 1-6 Alkyl, Halogenated C1-6 Alkyl and C 1-6 Alkoxy C 1-6 alkyl-;

[0020] Alternatively, a substituent on Y and Q 3 The substituents on them are linked together to form a macrocyclic structure.

[0021] In some implementation schemes, X 1 Let N, X 2 For NR 4 ,as well as For a bridge ring, where R 1 It is hydrogen, hydroxyl, or optionally halogenated, hydroxyl, C 1-6 Alkoxy, deuterated C 1-6 Alkoxy, -NR 1a R 1b C replaced by deuterium 1-6 alkyl.

[0022] In some implementation schemes, X 1 Let N, X 2 For NR 4 ,as well as for Where n is an integer between 2 and 4, and R 1 It is hydrogen, hydroxyl, or optionally halogenated, hydroxyl, C 1-6 Alkoxy, deuterated C 1-6 Alkoxy, -NR 1a R 1b C replaced by deuterium 1-6 alkyl.

[0023] In some implementation schemes, Y is Where R 1 For hydrogen, hydroxyl, C 1-6 Alkyl, Halogenated C 1-6 Alkyl, deuterated C 1-6 Alkyl, C 1-6 Alkoxy C 1-6 Alkyl- or deuterated C 1-6 Alkoxy C 1-6 alkyl-.

[0024] In some implementations, Y is

[0025] In some implementations, L is -O- or -O-(CH2). t -or Where t is an integer from 1 to 6, t1+t2 is an integer from 0 to 5, and t3 is an integer from 1 to 3. In some implementations, L is -O-CH2- or -O-.

[0026] In some implementation schemes, Q 1 and M 1 All are N.

[0027] In some implementation schemes, Q 3 and M 2 All are CH. In some implementations, Q 2 Let N be the number of elements in the array.

[0028] In some implementation schemes, Q 1 and M 1 Both are N, Q 3 and M 2 Both are CH, and Q 2 Let N be the number of elements in the array.

[0029] In some implementation schemes, R 2 The heterocyclic group is unsubstituted or modified by deuterium, halogen, or C. 1-6 Alkyl, C 1-6 Alkylene, Halogenated C 1-6 Alkylene, -OR 2a and -(CR 2a R 2b ) m -OC(O)NR 2c R 2d One or more substitutions in the heterocyclic group, wherein each variable is defined as in equation (I); preferably, the heterocyclic group is unsubstituted or substituted with deuterium, halogen, or C. 1-6 Alkyl, C 1-6 Alkylene, Halogenated C 1-6 Alkylene and -OR 2a One or more substitutions in the heterocyclic group; more preferably, the heterocyclic group is unsubstituted or substituted with one or two of deuterium, halogen, methyl, methoxy, methylene, or halomethylene. In a further embodiment, R 2 It is a 4- to 8-membered monocyclic heterocycle containing 1, 2, or 3 heteroatoms selected from oxygen, nitrogen, and sulfur as ring members, or a 6- to 12-membered bicyclic heterocycle (preferably a bridged bicyclic) containing 1, 2, or 3 heteroatoms selected from oxygen, nitrogen, and sulfur as ring members, wherein the monocyclic or bicyclic heterocyclic group is unsubstituted or substituted with deuterium, halogen, or C. 1-6 Alkyl, C 1-6 Alkylene, Halogenated C 1-6 Alkylene, -OR 2a and -(CR 2a R 2b ) m -OC(O)NR 2c R 2d One or more substitutions in the heterocyclic group, wherein each variable is defined as in equation (I); preferably, the heterocyclic group is unsubstituted or substituted with deuterium, halogen, or C.1-6 Alkyl, C 1-6 Alkylene, Halogenated C 1-6 Alkylene and -OR 2a One or more substitutions in the heterocyclic group; more preferably, the heterocyclic group is unsubstituted or substituted with one or two of deuterium, halogen, methyl, methoxy, methylene, or halomethylene. In a further embodiment, R 2 It is a monocyclic heterocyclic ring, which is piperidinyl, aza-butyl, or pyrrolidinyl, wherein the ring is unsubstituted or converted by one or two halogens, C 1-6 Alkyl, C 1-6 Alkylene or halogenated C 1-6 Alkyl-derivatization. In a further embodiment, R... 2 It is a bicyclic heterocycle, namely octahydrocyclopentadiene, wherein at least one carbon atom is replaced by a nitrogen atom, and one of the other carbon atoms is optionally replaced by an oxygen atom.

[0030] In some implementations, L is -O-CH2- or and R 2 The heterocyclic group is unsubstituted or halogenated, C 1-6 Alkyl, C 1-6 Alkylene, Halogenated C 1-6 Alkylene, -OR 2a and -(CR 2a R 2b ) m -OC(O)NR 2c R 2d One or more substitutions in the heterocyclic group, wherein each variable is defined as in equation (I); preferably, the heterocyclic group is unsubstituted or halogenated, C 1-6 Alkyl, C 1-6 Alkylene, Halogenated C 1-6 Alkylene and -OR 2a One or more substitutions in the heterocyclic group; more preferably, the heterocyclic group is unsubstituted or substituted with one or two of a halogen, methyl, methoxy, methylene, or halomethylene group. In a further embodiment, L is -O-CH2- or and R 2 It is a 4- to 8-membered monocyclic heterocycle containing 1, 2, or 3 heteroatoms selected from oxygen, nitrogen, and sulfur as ring members, or a 6- to 12-membered bicyclic heterocycle (preferably a bridged bicyclic) containing 1, 2, or 3 heteroatoms selected from oxygen, nitrogen, and sulfur as ring members, wherein the monocyclic or bicyclic heterocyclic group is unsubstituted or converted by halogen, C 1-6 Alkyl, C 1-6 Alkylene, Halogenated C 1-6 Alkylene, -OR 2a and -(CR 2a R 2b )m -OC(O)NR 2c R 2d One or more substitutions in the heterocyclic group, wherein each variable is defined as in equation (I); preferably, the heterocyclic group is unsubstituted or halogenated, C 1-6 Alkyl, C 1-6 Alkylene, Halogenated C 1-6 Alkylene and -OR 2a One or more substitutions in the heterocyclic group; more preferably, the heterocyclic group is unsubstituted or substituted with one or two of halogen, methyl, methoxy, methylene, and halomethylene. In a further embodiment, L is -O-CH2- or and R 2 It is a monocyclic heterocyclic ring, which is piperidinyl, aza-butyl, or pyrrolidinyl, wherein the ring is unsubstituted or converted by one or two halogens, C 1-6 Alkyl, C 1-6 Alkylene, Halogenated C 1-6 Alkyl-substituted. In a further embodiment, L is -O-CH2- or and R 2 It is a bicyclic heterocycle, namely octahydrocyclopentadiene, wherein at least one carbon atom is replaced by a nitrogen atom, and one of the other carbon atoms is optionally replaced by an oxygen atom.

[0031] In a further preferred embodiment, LR 2 for

[0032] In some implementation schemes, R 3 It is an aryl or heteroaryl group of a benzo5-8 membered saturated ring or heterocycle, wherein the benzo5-8 membered saturated ring, aryl or heteroaryl group is substituented by 1 to 3 R groups. 8 Replace, R 8 Each element is independently selected from: hydrogen, halogens, or C elements optionally substituted with halogens. 1-6 Alkyl, C2-C6 ynyl, -P(=O)R 8a R 8b -NR 8a R 8b -OR 8a -SR 8a Optional halogenated or C 1-6 Alkyl-substituted C3-C6 cycloalkyl, wherein R 8a and R 8b Each independent hydrogen, -C(O)OC 1-6 Alkyl groups and -C(O)NHC 1-6 Alkyl group. In some embodiments, R 3 The benzo5-8 membered saturated ring is composed of 1 to 3 substituents R.8 Replace, R 8 Each element is independently selected from: hydrogen, halogens, or C elements optionally substituted with halogens. 1-6 alkyl.

[0033] In some implementation schemes, R 3 for

[0034] Where: R 8 Preferred from -OH, -NH2, -OC(O)-C 1-6 Alkyl group, -OC(O)CH2CH3, -OC(O)NH-C 1-6 Alkyl or alkoxy.

[0035] In some embodiments, the compound of formula (I) is as shown in formula (I-1):

[0036] in

[0037] Y is

[0038] R 6 It is hydrogen, ethynyl, propynyl, methoxy, isopropoxy, or halogenated C. 1-6 Alkyl and C 1-6 Alkoxy;

[0039] R 7 It is hydrogen, halogen, methoxy, -OCD3, halogenated or unsubstituted C 1-6 Alkoxy;

[0040] LR 2 for The remaining variables are as defined in equation (I).

[0041] In some embodiments, the compound of formula (I) is as shown in formula (I-1-1):

[0042] in,

[0043] R 1 The derivatives are hydrogen, methyl, ethyl, -CH2-OH, -CH2-OCD3, CD3, and CD2CD3.

[0044] R 6 It can be hydrogen, ethynyl, propynyl, methoxy, -OCD3, halogenated or unsubstituted C 1-6 Alkoxy;

[0045] R 7 Preferably, it contains hydrogen, halogen, methoxy, -OCD3, halogenated or unsubstituted C. 1-6Alkoxy;

[0046] LR 2 for

[0047] R 3 for

[0048] The remaining variables are as defined in equation (I).

[0049] In some implementations, Q in compound (I) 3 For CR 6 And R 6 With R 1 The connection forms a new 7-8 quintic heterocycle, as shown in equation (I-2):

[0050] Among them: G 1 and G 2 Each of these is independent: single bond, -O-, -S-, -NR La -、-O-(CR La R Lb ) t -、-S-(CR La R Lb ) t -、-NR c -(CR La R Lb ) t -、-(CR La R Lb ) t -O-、-(CR La R Lb ) t -S-、-(CR La R Lb ) t -NR Lc -, -C(O)-, -SO2-, -SO-, -C(O)-O-, -OC(O)-, -C(O)-NR Lc -or-N Lc C(O)-, where R La R Lb and R Lc Each is independently selected from hydrogen and deuterated C. 1-6 Alkyl or C 1- 6 alkyl groups, and t is an integer from 1 to 6, with the remaining variables as defined for formula (I).

[0051] In some embodiments, the compound of formula (I) is as shown in formula (I-2-1):

[0052] Among them: G 1 and G 2 Each of these is independent: single bond, -O-, -S-, -NR La -、-O-(CR La R Lb ) t -、-S-(CR La R Lb ) t -、-NR c -(CR La R Lb ) t -、-(CR La R Lb ) t -O-、-(CR La R Lb ) t -S-、-(CR La R Lb ) t -NR Lc -, -C(O)-, -SO2-, -SO-, -C(O)-O-, -OC(O)-, -C(O)-NR Lc -or-N Lc C(O)-, where R La R Lb and R Lc Each is independently selected from hydrogen and deuterated C. 1-6 Alkyl or C 1- 6 alkyl groups, wherein t is an integer from 1 to 6, R 1 Preferably, the constituents are hydrogen, methyl, ethyl, -CH2-OH, -CH2-OCD3, CD3, and CD2CD3, Q. 2 Preferably N or CR 6 The remaining variables are as defined in equation (I).

[0053] In some embodiments, the compound of formula (I) is as shown in formula (I-2-1):

[0054] Among them: G 1 -G 2 -O-(CR) La R Lb ) t -、-S-(CR La R Lb ) t -、-NR c -(CR La R Lb ) t -、-(CRLa R Lb ) t -O-、-(CR La R Lb ) t -S- or -(CR) La R Lb ) t -NR Lc -, where R La R Lb and R Lc Each is independently selected from hydrogen and deuterated C. 1-6 Alkyl or C 1-6 Alkyl groups, and wherein t is an integer from 1 to 6, R 1 Preferably, it is hydrogen, methyl, ethyl, -CH2-OCD3, CD3, and CD2CD3, Q 2 Preferably N or CR 6 The remaining variables are as defined in equation (I). In some implementations, G 1 -G 2 -O-(CHR) La )-、-S-(CHR La )-、-NH-(CHR La )-、-(CHR La )-O-、-(CHR La -S- or -(CHR) La )-NH-, where R La Selected from hydrogen and deuterated C 1-6 Alkyl or C 1-6 Alkyl, R 1 The preferred components are hydrogen, methyl, ethyl, -CH2-OH, -CH2-OCD3, CD3, and CD2CD3, with the remaining variables as defined for formula (I).

[0055] In some embodiments, the compound of formula (I) is shown as in formula (I-2-2).

[0056] Where: n is an integer from 1 to 6, G 1 and G 2 Each of these is independent: single bond, -O-, -S-, -NR La -、-O-(CR La R Lb ) t -、-S-(CR La R Lb ) t -、-NR c -(CR La R Lb ) t -、-(CRLa R Lb ) t -O-、-(CR La R Lb ) t -S-、-(CR La R Lb ) t -NR Lc -, -C(O)-, -SO2-, -SO-, -C(O)-O-, -OC(O)-, -C(O)-NR Lc -or-N Lc C(O)-, where R La R Lb and R Lc Each is independently selected from hydrogen and deuterated C. 1-6 Alkyl or C 1-6 Alkyl groups, and wherein t is an integer from 1 to 6, R 1 Preferably, the constituents are hydrogen, methyl, ethyl, -CH2-OH, -CH2-OCD3, CD3, and CD2CD3, Q. 2 Preferably N or CR 6 The remaining variables are as defined in equation (I).

[0057] In some embodiments, the compound of formula (I) is as shown in formula (I-2-3).

[0058] Where: n is an integer from 1 to 6, G 1 and G 2 Each of these is independent: single bond, -O-, -S-, -NR La -、-O-(CR La R Lb ) t -、-S-(CR La R Lb ) t -、-NR c -(CR La R Lb ) t -、-(CR La R Lb ) t -O-、-(CR La R Lb ) t -S-、-(CR La R Lb ) t -NR Lc -, -C(O)-, -SO2-, -SO-, -C(O)-O-, -OC(O)-, -C(O)-NR Lc -or-NLc C(O)-, where R La R Lb and R Lc Each is independently selected from hydrogen and deuterated C. 1-6 Alkyl or C 1-6 Alkyl groups, and wherein t is an integer from 1 to 6, R 1 Preferably, hydrogen, methyl, ethyl, trifluoromethyl, trifluoroethyl, C 3-6 Cycloalkyl, -CH2-OCD3, CD3 and CD2CD3, Q 2 Preferably N or CR 6 The remaining variables are as defined in equation (I).

[0059] In some implementations, the compound of formula (I) is

[0060] It will be readily apparent to those skilled in the art that the compounds of the present invention, such as those of formula (I) or specific compounds, can be prepared according to one or more methods or in other ways known in this art. It is obvious that, in general, when following the general route described in this application, it is necessary to use starting materials with different substitutions and / or protecting groups to obtain the desired compounds. Different substituents can also be added at different points in the synthetic route to prepare the desired compounds.

[0061] This invention relates to pharmaceutical compositions of compounds of formula (I) or pharmaceutically acceptable salts, prodrugs and solvates thereof.

[0062] Another aspect of the present invention provides a method for treating disease symptoms using the compounds or pharmaceutical compositions of the present invention, including but not limited to symptoms related to G12 K-RAS, H-RAS, or N-RAS mutations (e.g., cancer), said method comprising administering a therapeutically effective amount of the compounds or pharmaceutical compositions of the present invention to a subject in need. Cancer mediated by KRas mutations and KRAS amplification is selected from pancreatic cancer, colon cancer, rectal cancer, lung adenocarcinoma, lung cancer, bile duct cancer, endometrial cancer, ovarian cancer, and leukemia; most preferably selected from pancreatic cancer, colon cancer, rectal cancer, lung adenocarcinoma, and bile duct cancer. Cancers mediated by G12C, G12D, G12V, G12A, G12R, G12S, G13D, and Q61H mutations include pancreatic cancer, lung cancer, colorectal cancer, etc., mediated by G12D mutations.

[0063] Another aspect of the invention provides the use of a compound or a pharmaceutically acceptable salt, tautomer, prodrug, or stereoisomer thereof in the preparation of a treatment for cancers associated with H-ras, K-ras, or N-ras inhibition.

[0064] The present invention relates to compounds of formula (I) which have good physicochemical properties and safety and toxicity parameters, and can be used for the treatment of cancer and inflammation in mammals.

[0065] In other embodiments, a method for inhibiting cell population proliferation is also provided, the method comprising contacting the cell population with any one of the compounds of structure (I).

[0066] Other embodiments involve pharmaceutical compositions. The pharmaceutical composition comprises any one (or more) of the aforementioned compounds and a pharmaceutically acceptable carrier. In some embodiments, the pharmaceutical composition is formulated for oral administration. In other embodiments, the pharmaceutical composition is formulated for injection. In still other embodiments, the pharmaceutical composition comprises a compound disclosed in this application and another therapeutic agent (e.g., an anticancer agent). Non-limiting examples of such therapeutic agents are described below.

[0067] Suitable routes of administration include, but are not limited to, oral, intravenous, rectal, aerosol, non-enteral, ocular, pulmonary, mucosal, percutaneous, vaginal, ear, nasal, and local administration. Additionally, by way of example only, non-enteral delivery includes intramuscular, subcutaneous, intravenous, intramedullary injection, as well as intrathecal, direct intracardiac, intraperitoneal, intralymphatic, and intranasal injection. Detailed Implementation

[0068] Unless otherwise specified, all disclosures of this invention are defined using the following terms:

[0069] The term "prodrug" refers to any derivative that can be converted into a corresponding active pharmaceutical compound in vivo. The prodrugs of the compounds described in this application readily undergo chemical changes under physiological conditions to be converted into the compounds of this invention. Furthermore, prodrugs can be converted into the compounds of this invention in the in vivo environment by chemical or biochemical methods.

[0070] The term "pharmaceutically acceptable salt" includes, unless otherwise stated, salts of acidic groups (e.g., but not limited to, potassium, sodium, magnesium, calcium, etc.) or salts of basic groups (e.g., but not limited to, formate, acetate, citrate, tartrate, methanesulfonate, malate, sulfate, hydrochloride, phosphate, nitrate, carbonate, etc.) that may be present in the compounds of the present invention.

[0071] The term "solvent" refers to a complex molecular compound formed in solution by solute molecules or ions attracting adjacent solvent molecules through intermolecular forces such as Coulomb forces, van der Waals forces, charge transfer forces, and hydrogen bonds. In one embodiment, the solvent is water, meaning the compound of this invention forms a hydrate.

[0072] The compounds of this invention, or pharmaceutically acceptable salts thereof, may contain one or more step symmetry centers, and thus may produce enantiomers, diastereomers, and other stereoisomers, defined in terms of the absolute stereochemical configuration of the amino acid as (R)- or (S)-, or (D)- or (L)-. This invention is intended to include all such possible isomers, as well as their racemic and optically pure forms. Optically active (+) and (-), (R)- and (S)- or (D)- and (L)- isomers can be obtained using chiral synthesis or chiral preparation, or by resolution using conventional techniques (e.g., chromatography and fractional crystallization). Conventional techniques for preparing / separating individual enantiomers include chiral synthesis from suitable optically pure precursors, and resolution of racemic mixtures (or racemic mixtures of salts or derivatives) using, for example, chiral high-performance liquid chromatography (HPLC). This invention provides pure isomers and mixtures of isomers, methods of their preparation and uses, and compositions comprising them. For simplicity, it will be referred to as compound (I) below, which means both the pure optical isomer and, where appropriate, a mixture of isomers in different proportions.

[0073] The compounds of this invention can exist in specific forms. Unless otherwise stated, the terms "tautomer" or "tautomer form" refer to isomers of different functional groups in dynamic equilibrium at room temperature, capable of rapidly interconverting into each other. If tautomerization is possible (e.g., in solution), chemical equilibrium of the tautomer can be achieved. For example, proton tautomers (also called prototropic tautomers) include interconversions via proton migration, such as keto-enol isomerization and imine-enamine isomerization. Valence tautomers include interconversions involving the rearrangement of some bonding electrons.

[0074] The term "alkyl" in this application refers to a hydrocarbon group selected from straight-chain saturated hydrocarbon groups and branched-chain saturated hydrocarbon groups, containing 1 to 18 (e.g., 1 to 12, further, 1 to 10, even further, 1 to 8, 1 to 6, or 1 to 4) carbon atoms. Alkyl groups containing 1 to 6 carbon atoms (i.e., C...) 1-6 Examples of alkyl groups include, but are not limited to, methyl, ethyl, 1-propyl or n-propyl (“n-Pr”), 2-propyl or isopropyl (“i-Pr”), 1-butyl or n-butyl (“n-Bu”), 2-methyl-1-propyl or isobutyl (“i-Bu”), 1-methylpropyl or sec-butyl (“s-Bu”), 1,1-dimethylethyl or tert-butyl (“t-Bu”), 1-pentyl, 2-pentyl, 3-pentyl, 2-methyl-2-butyl, 3-methyl-2-butyl, 3-methyl-1-butyl, 2-methyl-1-butyl, 1-hexyl, 2-hexyl, 3-hexyl, 2-methyl-2-pentyl, 3-methyl-2-pentyl, 4-methyl-2-pentyl, 3-methyl-3-pentyl, 2-methyl-3-pentyl, 2,3-dimethyl-2-butyl, and 3,3-dimethyl-2-butyl.

[0075] The term "alkylene" in this application refers to a group formed by removing two hydrogen atoms from the same carbon atom of a straight-chain saturated hydrocarbon or a branched-chain saturated hydrocarbon, containing 1 to 18 (e.g., 1 to 12, further, 1 to 10, even further, 1 to 8, 1 to 6, or 1 to 4) carbon atoms. Alkylenes containing 1 to 6 carbon atoms (i.e., C...) 1-6 Examples of alkylene groups include, but are not limited to, methylene and ethylene.

[0076] The term "halogen" in this application refers to fluorine (F), chlorine (Cl), bromine (Br), and iodine (I).

[0077] The term "haloalkyl" in this application refers to an alkyl group in which one or more hydrogen atoms are replaced by one or more halogen atoms (such as fluorine (F), chlorine (Cl), bromine (Br), and iodine (I)). Examples of haloalkyl groups include halogenated C-type alkyl groups.1-8 Alkyl, Halogenated C 1-6 Alkyl or halogenated C 1-4 Alkyl groups, but not limited to -CF3, -CH2Cl, -CH2CF3, -CCl2, CF3, etc.

[0078] The term "alkenyl" in this application refers to a hydrocarbon group selected from straight-chain and branched hydrocarbon groups, which contains at least one C=C double bond and 2 to 18 (e.g., 2 to 8, more preferably 2 to 6) carbon atoms. Examples of alkenyl groups include C… 2-6 Alkenyl groups include, but are not limited to, ethenyl, propyl-1-enyl, propyl-2-enyl, 2-methylpropyl-1-enyl, but-1-enyl, but-2-enyl, but-3-enyl, but-1,3-dienyl, 2-methylbut-1,3-dienyl, hex-1-enyl, hex-2-enyl, hex-3-enyl, hex-4-enyl, and hex-1,3-dienyl.

[0079] The term "alkynyl" in this application refers to a hydrocarbon group selected from straight-chain and branched hydrocarbon groups, containing at least one C≡C triple bond and 2 to 18 (e.g., 2 to 8, further such as 2 to 6) carbon atoms. An example of an alkynyl group is C... 2-6 Alkynyl groups include, but are not limited to, ethynyl, 1-propynyl, 2-propynyl (propynyl), 1-butynyl, 2-butynyl and 3-butynyl.

[0080] In this application, the term "alkoxy" refers to an alkyl group as defined above that is bonded to oxygen, represented by -O alkyl. Examples of alkoxy groups include C0... 1-6 Alkoxy or C 1-4 Alkoxy groups include, but are not limited to, methoxy, ethoxy, isopropoxy, propoxy, n-butoxy, tert-butoxy, pentoxy, and hexoxy.

[0081] The term "cycloalkyl" in this application refers to a hydrocarbon group selected from saturated and partially unsaturated cyclic hydrocarbon groups, comprising monocyclic and polycyclic (e.g., bicyclic and tricyclic) groups. For example, a cycloalkyl group may contain 3 to 12 (e.g., 3 to 10, further, 3 to 8, further, 3 to 6, 3 to 5, or 3 to 4) carbon atoms. Even further, for example, a cycloalkyl group may be selected from a monocyclic group containing 3 to 12 (e.g., 3 to 10, further, 3 to 8, 3 to 6) carbon atoms. Examples of monocyclic cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, 1-cyclopent-1-enyl, 1-cyclopent-2-enyl, 1-cyclopent-3-enyl, cyclohexyl, 1-cyclohexyl-1-enyl, 1-cyclohexyl-2-enyl, 1-cyclohexyl-3-enyl, cyclohexadienyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl, cycloundecyl, and cyclododecyl. In particular, examples of saturated monocyclic cycloalkyl groups include C 3-8Cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. In a preferred embodiment, the cycloalkyl group is a monocyclic ring containing 3 to 6 carbon atoms (abbreviated as C). 3-6 Cycloalkyl groups, including but not limited to cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl. Examples of bicyclic cycloalkyl groups include those having 7 to 12 ring atoms arranged in a bicyclic arrangement selected from the [4,4], [4,5], [5,5], [5,6], or [6,6] ring systems, or bridging bicyclic groups selected from bicyclic [2.2.1]heptane, bicyclic [2.2.2]octane, and bicyclic [3.2.2]nonane. Other examples of bicyclic cycloalkyl groups include those bicyclic cycloalkyl groups arranged in a bicyclic arrangement selected from the [5,6] and [6,6] ring systems, such as... The wavy lines represent attachment points. The rings may be saturated or have at least one double bond (i.e., partially unsaturated), but are not fully conjugated and are not aromatic, as aromatic is defined in this application.

[0082] The term "aryl," used alone or in combination with other terms, refers to a group selected from the following:

[0083] a. 5- and 6-membered carbon ring aromatic rings, such as phenyl;

[0084] b. Bicyclic systems, such as 7- to 12-membered bicyclic systems, wherein at least one ring is a carbocyclic ring and aromatic, for example, a naphthyl group; and

[0085] c. Tricyclic systems, such as 10- to 15-membered tricyclic systems, in which at least one ring is a carbocyclic and aromatic, such as fluorene.

[0086] The terms "aromatic ring" and "aryl" are used interchangeably in the disclosure of this application. In some embodiments, the monocyclic or bicyclic aromatic ring has 5 to 10 cyclic carbon atoms (i.e., C10, C20, C30, C40, C50, C60, C7 ... 5-10 Aryl). Examples of monocyclic or bicyclic aromatic hydrocarbon rings include, but are not limited to, phenyl, naphth-1-yl, naphth-2-yl, anthraceneyl, phenanthrene, etc. In some embodiments, the aromatic hydrocarbon ring is a naphthyl ring (naphth-1-yl or naphth-2-yl) or a benzene ring. In some embodiments, the aromatic hydrocarbon ring is a benzene ring. In some embodiments, if a group as a whole is not aromatic, but its substitution position is located on an aromatic ring, then the group can be considered an aryl group, such as 5,6,7,8-tetrahydronaphth-1-yl, 5,6,7,8-tetrahydronaphth-2-yl, 5,6,7,8-tetrahydronaphth-3-yl, or 5,6,7,8-tetrahydronaphth-4-yl.

[0087] The term "heteroaryl" in this application refers to a group selected from the following:

[0088] a. A 5, 6, or 7-membered aromatic monocyclic ring containing at least one heteroatom, such as 1 to 4 heteroatoms, or in some embodiments 1 to 3 heteroatoms, or in some embodiments 1 to 2 heteroatoms, wherein the heteroatoms are selected from nitrogen (N), sulfur (S), and oxygen (O) (as one or more ring atoms), and the remaining ring atoms are carbon.

[0089] b. A 7- to 12-membered bicyclic ring comprising at least one heteroatom, for example, 1 to 4 heteroatoms, or in some embodiments, 1 to 3 heteroatoms, or in other embodiments, 1 or 2 heteroatoms selected from N, O, and S (as one or more ring atoms), the remaining ring atoms being carbon, and wherein at least one ring is aromatic and at least one heteroatom is present in the aromatic ring; and

[0090] c. A 11- to 14-membered tricyclic ring comprising at least one heteroatom, for example, 1 to 4 heteroatoms, or in some embodiments, 1 to 3 heteroatoms, or in other embodiments, 1 or 2 heteroatoms selected from N, O, and S (as one or more ring atoms), the remaining ring atoms being carbon, and wherein at least one ring is aromatic and at least one heteroatom is present in the aromatic ring.

[0091] In a preferred embodiment, the heteroaryl group is a 5- to 6-membered heteroaryl group containing one nitrogen atom and 0 or 1 additional heteroatoms selected from N, O and S, including but not limited to pyridyl, isoxazolyl and oxazolyl.

[0092] When the total number of S and O atoms in a heteroaryl group exceeds 1, those heteroatoms are not adjacent to each other. In some embodiments, the total number of S and O atoms in a heteroaryl group is no greater than 2. In some embodiments, the total number of S and O atoms in an aromatic heterocycle is no greater than 1. When a heteroaryl group contains more than one heteroatom ring member, the heteroatoms can be the same or different. The nitrogen atom in one or more rings of a heteroaryl group can be oxidized to form N-oxides.

[0093] The terms "aromatic heterocycle" and "heteroaryl" are used interchangeably in the disclosure of this application. In some embodiments, the monocyclic or bicyclic aromatic heterocycle has 5, 6, 7, 8, 9, or 10 cyclic members, wherein 1, 2, 3, or 4 heteroatomic ring members are independently selected from nitrogen (N), sulfur (S), and oxygen (O), and the remaining ring members are carbon. In some embodiments, the monocyclic or bicyclic aromatic heterocycle is a monocyclic or bicyclic ring comprising 1 or 2 heteroatomic ring members independently selected from nitrogen (N), sulfur (S), and oxygen (O). In some embodiments, the monocyclic or bicyclic aromatic heterocycle is a 5- to 6-membered heteroaryl ring, which is monocyclic and has 1 or 2 heteroatomic ring members independently selected from nitrogen (N), sulfur (S), and oxygen (O). In some embodiments, the monocyclic or bicyclic aromatic heterocycle is an 8- to 10-membered heteroaryl ring, which is bicyclic and has 1 or 2 heteroatomic ring members independently selected from nitrogen, sulfur, and oxygen.

[0094] Examples of heteroaryl or monocyclic or bicyclic aromatic heterocycles include, but are not limited to (numbered from the linking position specified as priority 1), pyridyl (e.g., 2-pyridyl, 3-pyridyl, or 4-pyridyl), cyclophosphino, pyrazinyl, 2,4-pyrimidinyl, 3,5-pyrimidinyl, 2,4-imidazolyl, imidazopyridyl, isoxazolyl, oxazolyl, thiazolyl, isothiazolyl, thiadiazolyl (e.g., 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, or 1,3,4-thiadiazolyl), tetrazolyl Thiophene (e.g., thiophene-2-yl, thiophene-3-yl), triazine, benzothiophene, furanyl (furyl or furanyl), benzofuranyl, benzimidazolyl, indole, isoyindolyl, dihydroindolyl, oxadiazolyl (e.g., 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl or 1,3,4-oxadiazolyl), phthalazinyl, pyrazinyl, pyridazinyl, pyrroleyl, triazolyl (e.g., 1,2,3-triazolyl, 1,2,4-triazolyl or 1,3,4-oxadiazolyl), phthalazinyl, pyrazinyl, pyridazinyl, pyrroleyl, triazolyl (e.g., 1,2,3-triazolyl, 1,2,4-triazolyl or 1,3,4-triazolyl), 4-triazolyl), quinolinyl, isoquinolinyl, pyrazolyl, pyrrolopyridyl (e.g., 1H-pyrrolo[2,3-b]pyridin-5-yl), pyrazolopyridyl (e.g., 1H-pyrazolo[3,4-b]pyridin-5-yl), benzoxazolyl (e.g., benzo[d]oxazol-6-yl), pteridinyl, purineyl, 1-oxa-2,3-diazolyl, 1-oxa-2,4-diazolyl, 1-oxa-2,5-diazolyl, 1-oxa-3,4-diazolyl, 1-thiazolyl -2,3-diazolyl, 1-thia-2,4-diazolyl, 1-thia-2,5-diazolyl, 1-thia-3,4-diazolyl, furazanyl (e.g., furazan-2-yl, furazan-3-yl), benzofurazanyl, benzobenzylthio, benzothiazolyl, benzoxazolyl, quinazolinyl, quinoxolinyl, naphthidyl, furan-pyridinyl, benzothiazolyl (e.g., benzo[d]thiazolyl-6-yl), indazole (e.g., 1H-indazole-5-yl), and 5,6,7,8-tetrahydroisoquinoline.

[0095] The terms “heterocyclic” or “heterocyclic” or “heterocyclic group” in this application refer to a ring selected from 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12-membered monocyclic, bicyclic and tricyclic saturated rings and partially unsaturated rings, which contains at least one carbon atom and at least one heteroatom, such as 1 to 4 heteroatoms, further such as 1 to 3 heteroatoms or further such as 1 or 2 heteroatoms, which are selected from nitrogen (N), sulfur (S), oxygen (O), -SO- or -SO2 (as one or more ring atoms).

[0096] In some embodiments, the heterocyclic group is a 4, 5, 6, 7, or 8-membered monocyclic ring having at least one heteroatom selected from N, O, and S. In some preferred embodiments, the heterocyclic group is a 4, 5, 6, 7, or 8-membered saturated monocyclic ring containing one nitrogen heteroatom. Exemplary heterocyclic groups are azirrocyclic butyl, pyrroliyl, piperidinyl, azirrocyclic heptyl, and azirrocyclic octyl. In other embodiments, the heterocyclic group is a 5, 6, 7, or 8-membered saturated monocyclic ring containing one nitrogen atom and one additional heteroatom selected from -NH, -O-, -S-, -SO-, or –SO2-. Exemplary heterocyclic groups are morpholino, morpholinyl, or piperazine rings. In some embodiments, the heterocyclic group is a 7 to 12-membered saturated bicyclic ring containing one nitrogen atom and 0, 1, or 2 additional heteroatoms selected from -NH, -O-, -S-, -SO-, or -SO2-. In some preferred embodiments, the heterocyclic group is a bicyclic bridging ring or a spirocyclic ring.

[0097] The term "heterocycle" in this application also refers to a 5- to 7-membered heterocycle fused with a 5, 6, and / or 7-membered alkyl, carbocyclic aromatic, or heteroaromatic ring, comprising at least one heteroatom selected from N, O, and S, provided that the entire ring structure is non-aromatic. The heterocycle is not a heteroaryl group as defined in this application. In a preferred embodiment, the heterocyclic group is a 5- to 6-membered heterocyclic group comprising one nitrogen atom and 0 or 1 additional heteroatom selected from N, O, and S, including but not limited to pyrrole, dihydropyridine, morpholino, morpholino, and tetrahydropyranoyl.

[0098] Examples of heterocycles include, but are not limited to (numbered starting from the linking position designated as priority 1), 1-pyrrolidinyl, 2-pyrrolidinyl, 2,4-imidazolidinyl, 2,3-pyrazolyl, 1-piperidinyl, 2-piperidinyl, 3-piperidinyl, 4-piperidinyl, 2,5-piperazinyl, pyranyl, morpholinyl, morpholino, 2-morpholinyl, 3-morpholinyl, ethylene oxide, acridineyl, cyclothioethane, azahexacyclic, oxacyclic, thiohexacyclic, 1,2-dithiohexacyclic, 1,3-dithiohexacyclic, dihydropyridinyl, tetrahydropyridinyl, thiomorpholinyl, thiaxyl, piperazinyl, homopiperazinyl, and homopiperazinyl. Piperidinyl, azirheptanyl, oxetaneheptyl, thiepanyl, 1,4-oxathianyl, 1,4-dioxetaneheptyl, 1,4-oxathianyl, 1,4-oxazetaneheptyl, 1,4-dithianyl, 1,4-azathianyl, oxazepinyl, diazathianyl diazepinyl), thiazepinyl, dihydrothiopheneyl, dihydropyranyl, dihydrofuranyl, tetrahydrofuranyl, tetrahydrothiopheneyl, tetrahydropyranyl, tetrahydrothiaranyl, 1-pyrrolizinyl, 2-pyrrolizinyl, 3-pyrrolizinyl, dihydroindolyl, 2H-pyranyl, 4H-pyranyl, 1,4-dioxane, 1,3-dioxolane, pyrazolinyl, pyrazolyl, dithianyl, dithiolanyl, pyrazolyl, imidazolinyl, pyrimidinone, 1,1-dioxo-thiomorpholinyl, 3-azabi Cyclo[3.1.0]hexyl, 3-azabicyclo[4.1.0]heptyl, azabicyclo[2.2.2]hexyl, tetrahydro-1H-pyrrolazin-7-yl, tetrahydro-1H-furano[3,4-b]pyrrole-3a-yl, tetrahydro-1H-furano[3,4-c]pyrrole-3a-yl, tetrahydro-1H-furano[3,4-b]pyrrole-6a-yl, octahydrocyclopentano[b]pyrrole-6a-yl, octahydrocyclopentano[c]pyrrole-3a-yl, octahydrocyclopentano[b]pyrrole-3a-yl, 3-azabicyclo[3.1.0]hexane-1-yl or 2-azabicyclo[3.1.0]hexane-1-yl. Substituted heterocycles also include cyclic systems substituted with one or more oxo groups, such as piperidinyl N-oxide, morpholino-N-oxide, 1-oxo-1-thiomorpholino, and 1,1-dioxo-1-thiomorpholino.

[0099] In some embodiments, the heterocyclic group is a non-aromatic fused bicyclic heterocyclic group, such as the fused bicyclic heterocycles listed above; and, for example, the following non-aromatic fused bicyclic heterocyclic groups.

[0100] "Optional" or "optionally" means that the event or condition described below may occur but is not required to occur, and the description includes both the scenario in which said event or condition occurs and the scenario in which said event or condition does not occur.

[0101] The term "substituted" means that any one or more hydrogen atoms on a particular atom are replaced by a substituent, which can include deuterium and hydrogen variants, provided that the valence state of the particular atom is normal and the substituted compound is stable. When the substituent is oxygen (i.e., =O), it means that two hydrogen atoms are replaced. Oxygen substitution does not occur on aromatic groups. The term "optionally substituted" means that it may or may not be substituted, unless otherwise specified, and the type and number of substituents can be arbitrary on a chemically feasible basis.

[0102] When any variable (e.g., R) appears more than once in the composition or structure of a compound, its definition is independent in each case. Thus, for example, if a group is substituted by 0-2 Rs, the group can optionally be substituted by at most two Rs, and R has independent options in each case. Furthermore, combinations of substituents and / or their variants are only permitted if such combinations produce a stable compound. The term "one or more substituents of the following groups" disclosed in this application includes, for example, 1 to 5 (e.g., 1 to 4, further such as 1, 2, or 3) substituents, provided the valence allows.

[0103] Unless otherwise specified, the term "heteroalkyl" on its own or in combination with another term refers to a stable straight-chain or branched alkyl group or combination thereof consisting of a certain number of carbon atoms and at least one heteroatom, or heterogroup. In some embodiments, the heteroatom is selected from B, O, N, and S, wherein the nitrogen and sulfur atoms are optionally oxidized, and the nitrogen heteroatom is optionally quaternized. In other embodiments, the heterogroup is selected from -C(=O)O-, -C(=O)-, -C(=S)-, -S(=O), -S(=O)2-, -C(=O)N(H)-, -N(H)-, -C(=NH)-, -S(=O)2N(H)-, and -S(=O)N(H)-. In some embodiments, the heteroalkyl is a C1-C6 heteroalkyl; in other embodiments, the heteroalkyl is a C1-C3 heteroalkyl. Heteroatoms or heteroatomic groups can be located in any internal position of a heteroalkyl group, including the position where the alkyl group is attached to the rest of the molecule, but the terms “alkoxy,” “alkamino,” and “alkthio” (or thioalkoxy) are conventional expressions referring to those alkyl groups that are attached to the rest of the molecule by an oxygen atom, an amino atom, or a sulfur atom, respectively. Examples of heteroalkyl groups include, but are not limited to, -OCH3, -OCH2CH3, -OCH2CH2CH3, -OCH2(CH3)2, -CH2-CH2-O-CH3, -NHCH3, -N(CH3)2, -NHCH2CH3, -N(CH3)(CH2CH3), -CH2-CH2-NH-CH3, -CH2-CH2-N(CH3)-CH3, -SCH3, -SCH2CH3, -SCH2CH2CH3, -SCH2(CH3)2, -CH2-SCH2-CH3, -CH2-CH2, -S(=O)-CH3, -CH2-CH2-S(=O)2-CH3, -CH=CH-O-CH3, -CH2-CH=N-OCH3, and -CH=CHNCCH3)-CH 3。 At most two heteroatoms can be consecutive, for example, -CH2-NH-OCH3.

[0104] Unless otherwise indicated, the alkyl, alkenyl, alkynyl, cycloalkyl, aryl, heterocyclic, and heteroaryl moieties described in this application may each be independently substituted by one or more groups selected from the following: hydroxyl, oxo, halogen, cyano, nitro, trifluoromethyl, azide, amino, carboxyl, and mercapto.

[0105] synthesis

[0106] Suitable solvents commonly used in organic reactions can be used in each step of the preparation method of the present invention, for example, but not limited to: aliphatic and aromatic, optionally hydrocarbons or halogenated hydrocarbons (e.g., pentane, hexane, heptane, cyclohexane, petroleum ether, gasoline, volatile oil, benzene, toluene, xylene, dichloromethane, dichloroethane, chloroform, carbon tetrachloride, chlorobenzene and o-dichlorobenzene), and aliphatic and aromatic, optionally alcohols (e.g., methanol, ethanol, propanol, isopropanol, tert-butyl...). Alcohols, ethylene glycol, etc.), ethers (e.g., diethyl ether and dibutyl ether, ethylene glycol dimethyl ether and diethylene glycol dimethyl ether, tetrahydrofuran and dioxane, etc.), esters (e.g., methyl acetate or ethyl acetate, etc.), nitriles (e.g., acetonitrile or propionitrile, etc.), ketones (e.g., acetone, butanone, etc.), amides (e.g., dimethylformamide, dimethylacetamide and N-methylpyrrolidone, etc.), as well as dimethyl sulfoxide, tetramethylene sulfone and hexamethylphosphoric triamine and N,N-dimethylpropane urea (DMPU), etc.

[0107] The following abbreviations are used in this invention: DCM represents dichloromethane; CHCl3 represents trichloromethane; EA represents ethyl acetate; THF represents tetrahydrofuran; MeCN represents acetonitrile; MeOH represents methanol; EtOH represents ethanol; i-PrOH represents isopropanol; PE represents petroleum ether; Toulene represents toluene; DMSO represents dimethyl sulfoxide; DMF represents N,N-dimethylformamide; DMA represents N,N-dimethylacetamide; CDCl3 represents deuterated chloroform; D2O represents heavy water; (CD3)2SO represents deuterated DMSO; CD3O D represents deuterated methanol; CuI represents cuprous iodide; DIPEA represents diisopropylethylamine; TEA represents triethylamine; K2CO3 represents potassium carbonate; Cs2CO3 represents cesium carbonate; Na2CO3 represents sodium carbonate; NaHCO3 represents sodium bicarbonate; NaOH represents sodium hydroxide; KOH represents potassium hydroxide; LiHMDS represents hexamethyldisilamide lithium; CDI replaces 1,1'-carbonylimidazole; MS represents mass spectrometry; NMR represents nuclear magnetic resonance; TFA represents trifluoroacetic acid; BINAP represents (2R,3S)-2,2'-diphenylphosphine-1.1'-Binaphthyl; BOC represents tert-butoxycarbonyl; Cbz represents benzyloxycarbonyl; DBU represents bicyclo-1,5-diaza-5-undecene; DCC represents 1,3-dicyclohexylcarbodiimide; DCE represents 1,2-dichloroethane; DMAP represents 4-dimethylaminopyridine; dppf represents bis(diphenylphosphino)ferrocene; LiAlH4 represents lithium aluminum hydride; LDA represents diisopropylaminolithium; m-CPBA represents m-chloroperoxybenzoic acid; MTM represents dimethyl sulfide; NBS represents N-bromosuccinimide; NCS represents N-chlorosuccinimide; NIS represents N-iodosuccinimide; PCC represents pyridine dichromate. TBAF represents tetrabutylamine fluoride; THP represents tetrahydropyranyl; TMEDA represents tetramethylethylenediamine; TMS represents trimethylsilyl; TMP represents 2,2,6,6-tetramethylpiperidine; Ts represents p-toluenesulfonyl; Pd(PPh3)4 represents tetratetraphenylphosphine palladium; PdCl2(dppf) represents 1,1'-bis(diphenylphosphine)ferrocene palladium dichloride; Pd2(dba)3 represents tridibenzylacetone dipalladium; HOBT represents 1-hydroxybenzotriazole; HATU represents 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate; TBTU represents O-benzotriazole-N, N,N',N'-Tetramethylurea tetrafluoroborate; Tf2O represents trifluoroacetic anhydride; Pd(OAc)2 represents palladium diacetate; RuPhos represents 2-dicyclohexylphosphine-2',6'-diisopropoxy-1,1'-biphenyl; Pd(PPh3)2Cl2 represents bis(triphenylphosphine)palladium dichloride; Sphos represents 3,2-bis(cyclohexylphosphine)-2',6'-dimethoxybiphenyl; XantPhos represents 4,5-bis(diphenylphosphine)-9,9-dimethyloxanthracene; MeONa represents sodium methoxide; n-BuLi represents n-butyllithium; t-BuONa represents sodium tert-butoxide; t-BuOK represents potassium tert-butoxide; KSCN represents... Potassium thiocyanate; CuBr represents cuprous bromide; NaNO2 represents sodium nitrite; Urea represents urea; POCl3 represents phosphorus oxychloride; BBr3 represents boron tribromide; NH4Cl represents ammonium chloride; MeI represents iodomethane; NMP represents N-methylpyrrolidone; K3PO4 represents potassium phosphate; column chromatography represents column chromatography separation; Ac represents acetyl; Bn represents benzyl; Fmoc represents fluorenylmethoxycarbonyl; Cy represents cyclohexyl; Tf represents trifluoromethanesulfonyl; cataCXium-A-Pd-G3 represents methanesulfonic acid [n-butyldi(1-adamantyl)phosphine](2-amino-1,1'-biphenyl-2-yl)palladium(II).

[0108] Synthesis of intermediate (S,E)-(4-(fluoromethylene)-1,3-dimethylpiperidin-3-yl)methanol:

[0109] Step 1: Synthesis of 1-(tert-butyl)3-ethyl(S)-4-((1-(tert-butoxy)-3-methyl-1-oxobutane-2-yl)amino)-5,6-dihydropyridine-1,3(2H)-dicarboxylate

[0110] Ethyl 1-N-Boc-4-oxo-3-piperidinic acid (13.8 g, 50.86 mmol) and tert-butyl (S)-2-amino-3-methylbutyrate (11.46 g, 66.12 mmol) were dissolved in anhydrous toluene (150 mL), followed by the addition of p-toluenesulfonic acid (0.88 g, 5.09 mmol). The mixture was then purged with nitrogen and refluxed under an oil-water separator for 12 h to remove the generated water. After the reaction was complete, the mixture was cooled to room temperature, concentrated to remove toluene, and then ethyl acetate and water were added. The organic phase was separated, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain a crude product. Column chromatography yielded a pale yellow oil. 21 g, yield: 96%. 1 H NMR(400MHz, CDCl3)δ9.09(d,J=9.4Hz,1H),4.27–3.96(m,4H),3.72(d,J=7.2Hz,1H),3.48(t,J=5.8Hz,2H) ,2.26(q,J=5.6Hz,2H),2.19–2.07(m,1H),1.44(d,J=4.6Hz,18H),1.25(t,J=7.0Hz,3H),1.03–0.92(m,6H).

[0111] Step 2: Synthesis of 1-(tert-butyl)-3-ethyl(R)-3-methyl-4-oxopiperidinium-1,3-dicarboxylate

[0112] Diisopropylamine (2.3 mL, 16.41 mmol) and anhydrous toluene (20 mL) were added to a three-necked flask, and nitrogen gas was introduced. After cooling to -78 °C in a dry ice-ethanol bath, n-butyllithium (2.5 M n-hexane solution, 6.56 mL, 16.41 mmol) was added dropwise to the reaction solution. After the addition was complete, the mixture was stirred at -78 °C for 40 min. Then, 1-(tert-butyl)3-ethyl(S)-4-((1-(tert-butoxy)-3-methyl-1-oxobutane-2-yl)amino)-5,6-dihydropyridine-1,3(2H)-dicarboxylate (5.0 g, 11.72 mmol) was dissolved in anhydrous toluene (10 mL) and added dropwise to the reaction solution. After stirring at -78 °C for 30 min, HMPA (2.94 g, 16.41 mmol) was added to the reaction solution, and stirring was continued for another 30 min. Iodomethane (1.09 mL, 17.58 mmol) was added to the reaction solution, and the solution was allowed to rise naturally to room temperature while stirring for 6 hours. After the reaction was complete, the reaction was quenched with 1 M hydrochloric acid (pH 4-5) under ice-water bath cooling. The mixture was extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to give a pale yellow oil. 1.57 g, yield: 45%. 1 H NMR (400MHz, CDCl3) δ4.52(d,J=13.6Hz,1H),4.18(tt,J=10.0,7.2,5.8Hz,3H),3.32(ddd,J=13.7,10.1,4.4Hz,1H),3.0 6(d,J=13.6Hz,1H),2.76(s,1H),2.48(dt,J=14.9,4.4Hz,1H),1.49(d,J=4.6Hz,9H),1.30(s,3H),1.26(t,J=7.1Hz,3H).

[0113] Step 3: Synthesis of 1-(tert-butyl)-3-ethyl(S,E)-4-(fluoromethylene)-3-methylpiperidine-1,3-dicarboxylate

[0114] (Fluoromethylene)triphenylphosphine tetrafluoroborate (3.01 g, 7.89 mmol) was dissolved in anhydrous tetrahydrofuran (15 mL), and the solution was purged with nitrogen three times. Under dry ice and ethanol conditions, the temperature of the reaction mixture was lowered to -70 °C, and a potassium tert-butoxide-tetrahydrofuran solution (7.89 mL, 1 M, 7.89 mmol) was added dropwise to the reaction system. The temperature was maintained and stirring continued for 1 h. Then, a solution of 1-(tert-butyl)-3-ethyl(R)-3-methyl-4-oxopiperidinium-1,3-dicarboxylate (1.5 g, 5.26 mmol) in anhydrous tetrahydrofuran (15 mL) was added dropwise to the reaction system. After the addition was complete, the resulting mixture was slowly heated to room temperature and stirred overnight. After the reaction was completed by TLC monitoring, the reaction solution was slowly poured into water (100 mL) and extracted three times with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain a colorless oil. 1.0g, yield: 63%. 1 H NMR (400MHz, CDCl3) δ6.55 (d, J=84.7Hz, 1H), 4.34 (s, 1H), 4.15 (s, 2H), 2.91 (ddd, J=12.8, 11.0, 3.9 Hz,1H),2.76(d,J=13.2Hz,1H),2.66(d,J=14.5Hz,1H),2.19(s,1H),1.46(s,9H),1.31–1.22(m,7H).

[0115] Step 4: Synthesis of (S,E)-4-(fluoromethylene)-3-methylpiperidine-3-carboxylic acid ethyl ester hydrochloride

[0116] 1-(tert-butyl)-3-ethyl(S,E)-4-(fluoromethylene)-3-methylpiperidine-1,3-dicarboxylate (1.0 g, 3.32 mmol) was added to 10 mL of 4 M dioxane hydrochloride solution and stirred at room temperature for 1 h. After the reaction was complete, the reaction solution was concentrated to obtain a solid, which was then slurried with 5 mL of tert-butyl methyl ether for 10 minutes. The mixture was filtered, the solid was collected, and dried under vacuum to obtain a white solid. 787 mg, yield: 100%. 1 H NMR (400MHz, DMSO) δ9.71(s,1H),8.51(s,1H),7.00(dd,J=83.7,1.7Hz,1H),4.17(qq,J=10.8,7.1Hz,2H),3.58(dd ,J=12.8,2.1Hz,1H),3.27–3.16(m,1H),2.90–2.73(m,3H),2.06–1.93(m,1H),1.29(s,3H),1.21(t,J=7.1Hz,3H).

[0117] Step 5: Synthesis of (S,E)-4-(fluoromethylene)-1,3-dimethylpiperidine-3-carboxylic acid ethyl ester

[0118] (S,E)-4-(fluoromethylene)-3-methylpiperidine-3-carboxylic acid ethyl ester hydrochloride (787 mg, 3.32 mmol) was dissolved in methanol (5 mL). Triethylamine was added dropwise until the pH of the reaction solution reached approximately 10. After stirring for 10 minutes, glacial acetic acid was added to the reaction solution until the pH reached approximately 4. A 37% aqueous formaldehyde solution (0.74 mL, 9.96 mmol) was added, and the mixture was stirred at room temperature for 30 minutes. Sodium cyanoborohydride (229 mg, 3.64 mmol) was added to the reaction solution, and the mixture was stirred at room temperature for 1 hour. After the reaction was complete as monitored by TLC, the reaction solution was concentrated, and column chromatography was used to separate the product into an oily substance. 706 mg, yield: 99%.

[0119] Step 6: Synthesis of (S,E)-(4-(fluoromethylene)-1,3-dimethylpiperidin-3-yl)methanol

[0120] Ethyl (S,E)-4-(fluoromethylene)-1,3-dimethylpiperidine-3-carboxylate (706 mg, 3.32 mmol) was dissolved in anhydrous tetrahydrofuran (5 mL). LiAlH4 (189 mg, 4.98 mmol) was added in portions to the reaction mixture while cooling in an ice-water bath. The reaction was stirred at room temperature for 30 min. TLC was used to monitor the reaction until complete. The reaction was quenched by adding sodium sulfate decahydrate. The mixture was filtered and washed with anhydrous tetrahydrofuran. The filtrate was dried over anhydrous sodium sulfate and concentrated to give a pale yellow oil. Yield: 92% (530 mg). 1 H NMR(400MHz, CDCl3) δ6.50(dd,J=86.4,2.4Hz,1H),3.78(dd,J=10.4,1.6Hz,1H),3.54(dd,J=10.6,1.8Hz ,1H),2.91–2.84(m,1H),2.77(ddd,J=13.7,4.0,2.1Hz,2H),2.23(s,3H),2.03–1.88(m,3H),0.92(s,3H).

[0121] Synthesis of intermediate (E)-2-(8-(fluoromethylene)-3-(methoxymethoxy)-5,6,7,8-tetrahydronaphth-1-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborane:

[0122] Step 1: Synthesis of (E)-8-bromo-1-(fluoromethylene)-6-(methoxymethoxy)-1,2,3,4-tetrahydronaphthalene

[0123] A solution of triphenylphosphine (229 mg, 0.6 mmol) in tetrahydrofuran (2 mL) was cooled to -70 °C, and a solution of potassium tert-butoxide in tetrahydrofuran (0.6 mL, 1 M) was added dropwise. The mixture was stirred at this temperature for 50 minutes, followed by the addition of a solution of 8-bromo-6-(methoxymethoxy)-3,4-dihydronaphthyl-1(2H)-one (114 mg, 0.4 mmol) in tetrahydrofuran (2 mL). The reaction solution was gradually heated to room temperature and stirred overnight. After the reaction was complete, the mixture was extracted with saturated ammonium chloride aqueous solution (10 mL) and ethyl acetate (10 mL). The organic phase was separated and washed successively with saturated sodium bicarbonate aqueous solution (5 mL) and saturated brine (5 mL), dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. Preparative TLC separation yielded an oil-free product (65 mg, yield: 54%). 1 H NMR (400MHz, CDCl3) δ7.37(d,J=44Hz,1H),7.18(d,J=4Hz,1H),6.93(d,J=4H z,1H),5.21(s,2H),3.37(s,3H),2.58(t,J=8Hz,2H),1.67(tt,J=4,8Hz,2H).

[0124] Step 2: Synthesis of (E)-2-(8-(fluoromethylene)-3-(methoxymethoxy)-5,6,7,8-tetrahydronaphth-1-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborane

[0125] A tetrahydrofuran solution (1 mL) of compound (E)-8-bromo-1-(fluoromethylene)-6-(methoxymethoxy)-1,2,3,4-tetrahydronaphthalene (60 mg, 0.2 mmol) was cooled to -68 °C, and a hexane solution of n-butyllithium (2.5 M, 0.11 mL) was added dropwise. The resulting brownish-yellow solution was stirred at this temperature for 40 minutes. Then, a tetrahydrofuran solution (1 mL) of isopropanol pinacol borate (52 mg, 0.28 mmol) was added, and the reaction mixture was slowly heated to room temperature and stirred for 8 hours. The reaction was quenched dropwise with a saturated ammonium chloride aqueous solution (1 mL), followed by extraction with ethyl acetate (10 mL) and water (10 mL) to separate the organic phase. The aqueous phase was extracted with ethyl acetate (5 mL). The combined organic phases were washed successively with a saturated sodium bicarbonate aqueous solution (5 mL) and a saturated brine solution (5 mL), dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. The target product was obtained by separation using preparative TLC plates as a colorless gel solid (26 mg, 0.043 mmol, yield 22%).

[0126] Example 1: Synthesis of 4-(4-((1R,5S)-3,8-diazabicyclo[3.2.1]octane-3-yl)-8-fluoro-2-(((S,E)-4-(fluoromethylene)-1,3-dimethylpiperidin-3-yl)methoxy)pyrido[4,3-d]pyrimidin-7-yl)-5-methyl-5,6,7,8-tetrahydronaphthol-2-phenol

[0127] Step 1: Synthesis of (1R,5S)-3-(7-chloro-8-fluoro-2-(((S,E)-4-(fluoromethylene)-1,3-dimethylpiperidin-3-yl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester

[0128] Compound (S,E)-(4-(fluoromethylene)-1,3-dimethylpiperidin-3-yl)methanol (526 mg, 3.04 mmol) was dissolved in anhydrous tetrahydrofuran (5 mL). 60% NaH (122 mg, 3.04 mmol) was added under ice-water bath cooling, and the mixture was stirred for 30 minutes under ice-water bath cooling. (1R,5S)-3-(2,7-dichloro-8-fluoropyridino[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester (1.0 g, 2.33 mmol) was added to the reaction solution, and the mixture was slowly brought to room temperature and stirred for 12 hours. After the reaction was complete as detected by TLC, saturated ammonium chloride was added to quench the reaction, followed by extraction with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain an off-white solid. Yield: 75% (1.0 g). ES + (m / z): 565.78 [M+H] + .

[0129] Step 2: Synthesis of (1R,5S)-3-(8-fluoro-2-(((S,E)-4-(fluoromethylene)-1,3-dimethylpiperidin-3-yl)methoxy)-7-(3-(methoxymethoxy)-8-methyl-5,6,7,8-tetrahydronaphthyl)pyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate

[0130] The compound (1R,5S)-3-(7-chloro-8-fluoro-2-(((S,E)-4-(fluoromethylene)-1,3-dimethylpiperidin-3-yl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester (100 mg, 0.18 mmol), 2-(3-(methoxymethoxy)-8-methyl-5,6,7,8- Tetrahydronaphthyl-1-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborane (88 mg, 0.27 mmol), potassium phosphate (113 mg, 0.53 mmol), and cataCXium-A-Pd-G3 (15 mg, 0.02 mmol) were added to a reaction flask, followed by the addition of ethanol (3 mL) and water (1 mL). The mixture was purged with nitrogen three times, heated to 75 °C, and stirred for 6 hours. After the reaction was complete as monitored by TLC, it was cooled to room temperature, extracted with ethyl acetate and water, dried over anhydrous sodium sulfate, concentrated, and purified by TLC to obtain an off-white solid. Yield: 77% (100 mg). ES + (m / z): 735.27 [M+H] + .

[0131] Step 3: Synthesis of 4-(4-((1R,5S)-3,8-diazabicyclo[3.2.1]octane-3-yl)-8-fluoro-2-(((S,E)-4-(fluoromethylene)-1,3-dimethylpiperidin-3-yl)methoxy)pyrido[4,3-d]pyrimidin-7-yl)-5-methyl-5,6,7,8-tetrahydronaphthyl-2-phenol

[0132] The compound (1R,5S)-3-(8-fluoro-2-(((S,E)-4-(fluoromethylene)-1,3-dimethylpiperidin-3-yl)methoxy)-7-(3-(methoxymethoxy)-8-methyl-5,6,7,8-tetrahydronaphthyl)pyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (100 mg, 0.14 mmol) was dissolved in dichloromethane (3 mL), and then a 1,4-dioxane solution of HCl (1 mL, 4 M) was added. The mixture was stirred at room temperature for 1 hour. After the reaction was completed by TLC monitoring, the reaction was quenched with saturated sodium bicarbonate aqueous solution, extracted with dichloromethane, washed with the organic phase and brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the target product as an off-white solid. 70 mg, yield: 87%. 1H NMR (400MHz, DMSO) δ9.24(s,1H),9.07(s,1H),6.69(dd,J=86.8,1.8Hz,1H),6.56(d,J=2.6Hz,1H),6 .48(d,J=2.6Hz,1H),4.62(d,J=10.6Hz,1H),4.46(d,J=12.0Hz,1H),4.28(dd,J=17.5,11.2Hz,2H),3 .61(d,J=11.9Hz,1H),3.52(t,J=9.6Hz,3H),3.06(d,J=8.5Hz,1H),2.70(dd,J=16.3,10.6Hz,4H),2 .22(dd,J=15.2,10.0Hz,1H),2.14(s,3H),1.95–1.51(m,10H),1.10(s,3H),0.71(d,J=7.0Hz,3H).ES + (m / z): 592.01 [M+H] + .

[0133] Examples 2-64 were obtained by applying the preparation method of Example 1.

[0134] SFC resolution conditions for diastereomers: Column: Chiralcel OJ-3, 150×4.6mm ID, 3μm; Mobile phase: A: CO2 / B: IPA (0.1% IPAm, v / v); Flow rate: 2.5mL / min; Column temp.: 35℃; ABPR: 2000psi.

[0135] Example 74: Synthesis of 4-((5S,5aS,6R,9S)-1-fluoro-12-((S,E)-4-(fluoromethylene)-1,3-dimethylpiperidin-3-yl)methoxy)-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaza-6,9-methylnaphtho[1,8-ab]hept-2-yl)-5-ethyl-5,6,7,8-tetrahydronaphthol-2-phenol

[0136] Step 1: Synthesis of (1R,2S,5S)-2-((S)-1-((7-chloro-8-fluoro-4-hydroxy-2-(methylthio)pyrido[4,3-d]pyrimidin-5-yl)oxy)ethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester

[0137] Under ice-water bath cooling, sodium hydrogen (256 mg, 6.41 mmol) was added to a tetrahydrofuran (18 mL) solution of (1R,2S,5S)-2-((S)-1-hydroxyethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester (468 mg, 1.83 mmol). The reaction mixture was stirred at this temperature for 30 min, followed by the addition of 5,7-dichloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4-ol (465 mg, 1.66 mmol). The reaction mixture was slowly brought to room temperature and stirred overnight. The reaction was quenched by the addition of a small amount of saturated ammonium chloride aqueous solution. The reaction mixture was extracted with dichloromethane (50 mL) and water (50 mL) and separated. The aqueous phase was extracted with dichloromethane (50 mL). The combined organic phases were dried over sodium sulfate and rotary evaporated to give a yellow viscous product (800 mg) which was used directly for the next step. MS:(ESI)m / z=499.3[MH] - .

[0138] Step 2: Synthesis of tert-butyl(5S,5aS,6R,9S)-2-chloro-1-fluoro-5-methyl-12-(methylthio)-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaza-6,9-methylnaphtho[1,8-ab]heptane-14 carboxylic acid ester

[0139] A solution of (1R,2S,5S)-2-((S)-1-((7-chloro-8-fluoro-4-hydroxy-2-(methylthio)pyrido[4,3-d]pyrimidin-5-yl)oxy)ethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester (500 mg, 1.0 mmol) and DIEA (0.52 mL, 3.0 mmol) in dichloromethane (30 mL) was cooled in an ice-water bath. Phosphorus oxychloride (0.47 mL, 5.0 mmol) was added dropwise. The resulting yellow solution was stirred at room temperature for 2 hours. The mixture was cooled in an ice-water bath, and a saturated aqueous sodium bicarbonate solution was added, followed by stirring for 1 hour to completely quench the reaction. The mixture was extracted with dichloromethane (20 mL) and a saturated aqueous sodium bicarbonate solution (30 mL), resulting in two phases. The aqueous phase was extracted with dichloromethane (10 mL). The combined organic phases were washed with saturated brine (10 mL), dried over sodium sulfate, and then rotary evaporated to obtain a solid. Separation by rapid column chromatography (ethyl acetate: dichloromethane = 1:40) yielded a pale yellow solid (301 mg, 63% yield). MS: (ESI) m / z = 481.3 [MH] -

[0140] Step 3: Synthesis of tert-butyl(5S,5aS,6R,9S)-2-chloro-1-fluoro-5-methyl-12-(methylthio)-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaza-6,9-methylnaphtho[1,8-ab]heptane-14 carboxylic acid ester

[0141] Add tert-butyl(5S,5aS,6R,9S)-2-chloro-1-fluoro-5-methyl-12-(methylthio)-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaza-6,9-methylnaphtho[1,8-ab]heptane-14 carboxylate (480 mg, 1.0 mmol), 5-methyl-4-tetra,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl-5,6,7,8-tetrahydronaphthol-2-phenol (398 mg, 1.2 mmol), CataCXium APd G4 (111 mg, 0.15 mmol), and tripotassium phosphate (636 mg, 3.0 mmol) to a round-bottom flask, purging the argon atmosphere three times. Add a mixture of dioxane (10 mL) and water (2 mL) as a solvent. The mixture was stirred overnight at 70°C. After cooling to room temperature, it was extracted with ethyl acetate (30 mL) and water (30 mL). The aqueous phase was extracted with ethyl acetate (10 mL) after separation. The combined organic phases were washed with saturated brine (20 mL) and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure, and the crude product was separated by column chromatography (dichloromethane:methanol = 50:1) to give a yellow solid (580 mg, 0.89 mmol, yield 89%). MS: (ESI) m / z = 651.5 [M+H] +

[0142] Step 4: Synthesis of tert-butyl(5S,5aS,6R,9S)-1-fluoro-2-(3-(methoxymethoxy)-8-methyl-5,6,7,8-tetrahydronaphth-1-yl)-5-methyl-12-(methylsulfinyl)-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaza-6,9-methylnaphtho[1,8-ab]heptane-14 carboxylic acid ester

[0143] Under ice-water bath cooling, m-chloroperoxybenzoic acid (32 mg, 0.159 mmol) was added in a single batch to a 2 mL solution of tert-butyl(5S,5aS,6R,9S)-2-chloro-1-fluoro-5-methyl-12-(methylthio)-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaza-6,9-methylnaphtho[1,8-ab]heptane-14 carboxylic acid ester (94 mg, 0.145 mmol) in dichloromethane. The resulting solution was stirred in an ice-water bath for 20 minutes. It was then diluted with 15 mL of dichloromethane. The solution was washed successively with 10 mL of sodium sulfite aqueous solution, 10 mL of saturated sodium bicarbonate aqueous solution, and 5 mL of saturated brine. The organic phase was dried over sodium sulfate and then rotary evaporated to obtain a crude solid product, which was directly used in the next reaction step.

[0144] Step 5; Synthesis of tert-butyl(5S,5aS,6R,9S)-1-fluoro-12-((S,E)-4-(fluoromethylene)-1,3-dimethylpiperidin-3-yl)methoxy)-2-(3-(methoxy-methoxy)-8-methyl-5,6,7,8-tetrahydronaphthyl-1-yl)-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaza-6,9-methylnaphtho[1,8-ab]heptane-14 carboxylic acid ester

[0145] Under ice-water bath cooling, a tetrahydrofuran solution of potassium tert-butoxide (0.217 mL, 1 M, 0.217 mmol) was added dropwise to a toluene solution of tert-butyl(5S,5aS,6R,9S)-1-fluoro-2-(3-(methoxymethoxy)-8-methyl-5,6,7,8-tetrahydronaphthyl-1-yl)-5-methyl-12-(methylsulfinyl)-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaza-6,9-methylnaphtho[1,8-ab]heptane-14 carboxylic acid ester (not exceeding 0.145 mmol) and (S,E)-(4-(fluoromethylene)-1,3-dimethylpiperidin-3-yl)methanol (25 mg, 0.145 mmol). The reaction solution was stirred overnight at room temperature. Extracted with ethyl acetate (20 mL) and water (15 mL). The organic phase was washed with saturated brine, dried over sodium sulfate, and then rotary evaporated. The resulting solid was separated by silica gel slurry (methanol:dichloromethane = 1:40) to obtain a foamy solid product (40 mg, 0.054 mmol, yield 36%).

[0146] Step Six: Synthesis of 4-((5S,5aS,6R,9S)-1-fluoro-12-((S,E)-4-(fluoromethylene)-1,3-dimethylpiperidin-3-yl)methoxy)-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaza-6,9-methylnaphtho[1,8-ab]hept-2-yl)-5-ethyl-5,6,7,8-tetrahydronaphthol-2-phenol

[0147] A reaction flask containing tert-butyl(5S,5aS,6R,9S)-1-fluoro-12-((S,E)-4-(fluoromethylene)-1,3-dimethylpiperidin-3-yl)methoxy)-2-(3-(methoxy-methoxy)-8-methyl-5,6,7,8-tetrahydronaphthyl-1-yl)-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaza-6,9-methylnaphtho[1,8-ab]heptane-14 carboxylic acid ester (10 mg, 0.0129 mmol) was added to a pre-mixed solution of acetonitrile (1 mL) and 1,4-dioxane (1 mL) of hydrogen chloride. The resulting pale yellow solution was stirred at room temperature for 30 minutes. The solution was diluted with ethyl acetate (5 mL), and the resulting suspension was rotary evaporated to give a white solid. Extracted and separated by dichloromethane (15 mL) and saturated sodium bicarbonate (15 mL), the aqueous phase was extracted with dichloromethane (10 mL × 2). The combined organic phases were washed with saturated brine (10 mL), dried over sodium sulfate, and rotary evaporated. The crude product was separated by TLC (methanol:dichloromethane = 1:8) to obtain a white powder product (5.5 mg, 0.0087 mmol, yield 67%). 1 H NMR (400MHz, CDCl3) δ9.26 (s, 1H), 6.71 (d, J = 88Hz, 1H), 6.55 (s, 1H), 6.48 (s, 1H), 5.22 (d, J = 12Hz, 1H), 4.64 (d,J=12Hz,1H),4.30(d,J=12Hz,1H),4.24-4.20(m,1H),4.02-3.97(m,1H),3.85-3.81(m,1H),3.27-3.20(m, 3H),3.12-3.06(m,1H),2.73-2.69(m,4H),2.51-2.33(m,1H),2.17(s,3H),1.91-1.64(m,8H),1.56-1.52(m, 1H),1.45(d,J=4.0Hz,3H),1.32(d,J=16Hz,1H),1.12(s,3H),0.70(d,J=8.0Hz,3H)MS:(ESI)m / z=633.5[M+H] +

[0148] Examples 75-123 were obtained by applying the preparation method of Example 74.

[0149] SFC resolution conditions for diastereomers: Column: Chiralcel OJ-3, 150×4.6mm ID, 3μm; Mobile phase: A: CO2 / B: IPA (0.1% IPAm, v / v); Flow rate: 2.5mL / min; Column temp.: 35℃; ABPR: 2000psi.

[0150] Example 124: Synthesis of (S)-4-(8-fluoro-2-(((S,E)-4-(fluoromethylene)-1,3-dimethylpiperidin-3-yl)methoxy)-7-((R)-3-hydroxy-8-methyl-5,6,7,8-tetrahydronaphth-1-yl)pyrido[4,3-d]pyrimidin-4-yl)-6-methyl-1,4-oxazacycloheptane-6-ol

[0151] Step 1: (S)-4-(2,7-dichloro-8-fluoropyridino[4,3-d]pyrimidin-4-yl)-6-methyl-1,4-oxazetane-6-ol

[0152] 2,4,7-Trichloro-8-fluoropyridino[4,3-d]pyrimidine (2 g, 7.92 mmol) was dissolved in anhydrous dichloromethane (20 mL). DIEA (3.07 g, 23.77 mmol) and (S)-6-methyl-1,4-oxazacycloheptan-6-ol hydrochloride (1.46 g, 8.72 mmol) were added under dry ice-ethanol bath cooling. The mixture was stirred for 10 minutes under dry ice-ethanol bath cooling. After the reaction was complete, the reaction was quenched with saturated ammonium chloride aqueous solution, extracted with ethyl acetate, washed with saturated laboratory water, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography (DCM / EA = 3:1 to 2:1) to give 2.7 g of white solid, yield 98.2%. 1 H NMR (400MHz, DMSO) δ9.49 (s, 1H), 5.21 (s, 1H), 4.37 (d, J = 14.0Hz, 1H), 4.25 (d, J=14.7Hz,1H),4.08–3.90(m,3H),3.82(s,1H),3.63–3.47(m,2H),1.16(s,3H).

[0153] Step 2: (S)-4-(7-chloro-8-fluoro-2-(((S,E)-4-(fluoromethylene)-1,3-dimethylpiperidin-3-yl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)-6-methyl-1,4-oxazacycloheptan-6-ol

[0154] (S)-4-(2,7-dichloro-8-fluoropyridino[4,3-d]pyrimidin-4-yl)-6-methyl-1,4-oxazacycloheptan-6-ol (100 mg, 0.288 mmol) and (S,E)-(4-(fluoromethylene)-1,3-dimethylpiperidin-3-yl)methanol (64 mg, 0.374 mmol) were dissolved in anhydrous 1,4-dioxane (2 mL), followed by the addition of DIEA (112 mg, 0.864 mmol). The mixture was heated to 95 °C and stirred for 24 hours. After the reaction was complete, the mixture was cooled to room temperature, diluted with water, extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and purified by TLC (DCM / MeOH = 20 / 1) to give 36 mg of a white solid, yield: 25%. MS: (ESI) m / z = 484.38 [M+H] +

[0155] Step 3: (S)-4-(8-fluoro-2-(((S,E)-4-(fluoromethylene)-1,3-dimethylpiperidin-3-yl)methoxy)-7-((R)-3-(methoxymethoxy)-8-methyl-5,6,7,8-tetrahydronaphth-1-yl)pyrido[4,3-d]pyrimidin-4-yl)-6-methyl-1,4-oxazacycloheptane-6-ol

[0156] The compound (S)-4-(7-chloro-8-fluoro-2-(((S,E)-4-(fluoromethylene)-1,3-dimethylpiperidin-3-yl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)-6-methyl-1,4-oxazacycloheptan-6-ol (36 mg, 0.07 mmol), (R)-2-(3-(methoxymethoxy)-8-methyl-5,6,7,8-tetrahydronaphthyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborane (32 mg, 0.10 mmol), potassium phosphate (47 mg, 0.22 mmol), and CataCXium A Pd G4 (8 mg, 0.01 mmol) were used to replace argon three times. A mixed solvent of dioxane (10 mL) and water (2 mL) was added. The mixture was stirred overnight at 70 °C. The mixture was cooled to room temperature and extracted with ethyl acetate (30 mL) and water (30 mL). After separation, the aqueous phase was extracted with ethyl acetate (10 mL). The combined organic phases were washed with saturated brine (20 mL) and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure, and the crude product was purified by preparative TLC (dichloromethane:methanol = 20:1) to give a white solid (25 mg, yield 52%). MS: (ESI) m / z = 654.44 [M+H] +

[0157] Step 4: (S)-4-(8-fluoro-2-(((S,E)-4-(fluoromethylene)-1,3-dimethylpiperidin-3-yl)methoxy)-7-((R)-3-hydroxy-8-methyl-5,6,7,8-tetrahydronaphth-1-yl)pyrido[4,3-d]pyrimidin-4-yl)-6-methyl-1,4-oxazetane-6-ol

[0158] The compound (S)-4-(8-fluoro-2-(((S,E)-4-(fluoromethylene)-1,3-dimethylpiperidin-3-yl)methoxy)-7-((R)-3-(methoxymethoxy)-8-methyl-5,6,7,8-tetrahydronaphthyl)pyrido[4,3-d]pyrimidin-4-yl)-6-methyl-1,4-oxazetane-6-ol (25 mg, 0.04 mmol) was dissolved in acetonitrile (1 mL), and then 0.5 mL of 1,4-dioxane solution in 4N HCl was added. The mixture was stirred at room temperature for 20 minutes. After the reaction was complete, a saturated sodium bicarbonate aqueous solution was added to quench the reaction. The mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated to obtain a crude product. The crude product was purified by preparative TLC (DCM / MeOH = 15 / 1) to give a white solid (15 mg, yield 75%). MS: (ESI) m / z = 610.42 [M+H] + .

[0159] Examples 125-145 were obtained by applying the preparation method of Example 124.

[0160] Example 146: Synthesis of (R)-4-((S)-1-fluoro-12-(((S,E)-4-(fluoromethylene)-1,3-dimethylpiperidin-3-yl)methoxy)-5a,6,9,10-tetrahydro-5H,8H-4,7-dioxa-3,10a,11,13-tetraazanaphtho[1,8-ab]hepten-2-yl)-5-methyl-5,6,7,8-tetrahydronaphthol:

[0161] Step 1: Synthesis of (S)-5-((1,4-oxazacycloheptane-3-yl)methoxy)-7-chloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4-ol

[0162] (R)-(1,4-oxazacycloheptan-3-yl)methanol (200 mg, 1.52 mmol) was dissolved in anhydrous tetrahydrofuran (20 mL), and 60% sodium hydride (221 mg, 5.54 mmol) was added under ice-water bath cooling. After stirring for 30 minutes under ice-water bath cooling, 5,7-dichloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4-ol (388 mg, 1.39 mmol) was added to the reaction solution, and the reaction was stirred at room temperature for 2 hours. After the reaction was completed, saturated ammonium chloride was added to quench the reaction, and a solid precipitated. The solid was filtered, the filter cake was washed with water, and dried under vacuum to give an off-white solid (495 mg, 95% yield).

[0163] Step 2: (S)-2-chloro-1-fluoro-12-(methylthio)-5a,6,9,10-tetrahydro-5H,8H-4,7-dioxa-3,10a,11,13-tetraazanaphtho[1,8-ab]heptene

[0164] (S)-5-((1,4-oxazacycloheptane-3-yl)methoxy)-7-chloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4-ol (495 mg, 1.32 mmol) was dissolved in anhydrous acetonitrile (10 mL), followed by the addition of DIEA (324 mg, 2.51 mmol) and HATU (753 mg, 1.98 mmol). The mixture was stirred at room temperature for 2 hours. After the reaction was complete, water was added to quench the reaction, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to give an off-white solid (340 mg, 72% yield).

[0165] Step 3: Synthesis of (S)-2-chloro-1-fluoro-12-(methanesulfonyl)-5a,6,9,10-tetrahydro-5H,8H-4,7-dioxa-3,10a,11,13-tetraazanaphtho[1,8-ab]heptene

[0166] The compound (S)-2-chloro-1-fluoro-12-(methylthio)-5a,6,9,10-tetrahydro-5H,8H-4,7-dioxa-3,10a,11,13-tetraazanaphtho[1,8-ab]hepten (495 mg, 1.39 mmol) was dissolved in dichloromethane (10 mL), and m-chloroperoxybenzoic acid (555 mg, 3.06 mmol) was added. The mixture was stirred at room temperature for 2 hours. After the reaction was complete, the reaction was quenched with saturated sodium bicarbonate, extracted with dichloromethane, dried over anhydrous sodium sulfate, and concentrated to obtain a crude product which was directly used in the next step (539 mg, 100% yield).

[0167] Step 4: Synthesis of (S)-2-chloro-1-fluoro-12-(((S,E)-4-(fluoromethylene)-1,3-dimethylpiperidin-3-yl)methoxy)-5a,6,9,10-tetrahydro-5H,8H-4,7-dioxa-3,10a,11,13-tetraazanaphtho[1,8-ab]heptenene

[0168] Compounds (S)-2-chloro-1-fluoro-12-(methanesulfonyl)-5a,6,9,10-tetrahydro-5H,8H-4,7-dioxa-3,10a,11,13-tetraazanaphtho[1,8-ab]hepten (74 mg, 0.19 mmol) and (S,E)-(4-(fluoromethylene)-1,3-dimethylpiperidin-3-yl)methanol (43 mg, 0.25 mmol) were dissolved in anhydrous tetrahydrofuran. Sodium tert-butoxide (22 mg, 0.23 mmol) was added under ice-salt bath cooling, and the reaction was stirred for 2 hours under ice-salt bath cooling. After the reaction was completed, saturated ammonium chloride was added to quench the reaction, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, concentrated, and purified by TLC to give a white solid (40 mg, yield: 44%). MS: (ESI) m / z = 482.29 [M+H] + .

[0169] Step 5: Synthesis of (S)-1-fluoro-12-(((S,E)-4-(fluoromethylene)-1,3-dimethylpiperidin-3-yl)methoxy)-2-((R)-3-(methoxymethoxy)-8-methyl-5,6,7,8-tetrahydronaphth-1-yl)-5a,6,9,10-tetrahydro-5H,8H-4,7-dioxa-3,10a,11,13-tetraazanaphtho[1,8-ab]heptenene

[0170] Argon gas was replaced three times with (S)-2-chloro-1-fluoro-12-(((S,E)-4-(fluoromethylene)-1,3-dimethylpiperidin-3-yl)methoxy)-5a,6,9,10-tetrahydro-5H,8H-4,7-dioxa-3,10a,11,13-tetraazanaphtho[1,8-ab]hepten (40 mg, 0.08 mmol), (R)-2-(3-(methoxymethoxy)-8-methyl-5,6,7,8-tetrahydronaphth-1-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborane (33 mg, 0.10 mmol), potassium phosphate (53 mg, 0.25 mmol), and CataCXium A Pd G4 (9 mg, 0.001 mmol). A mixed solvent of dioxane (2 mL) and water (0.5 mL) was then added. The mixture was stirred overnight at 70°C. After cooling to room temperature, it was extracted with ethyl acetate (20 mL) and water (20 mL). The aqueous phase was extracted with ethyl acetate (10 mL) after separation. The combined organic phases were washed with saturated brine (20 mL) and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure, and the crude product was purified by preparative TLC (dichloromethane:methanol = 20:1) to give a white solid (35 mg, 64% yield). MS: (ESI) m / z = 652.61 [M+H] + .

[0171] Step 6: Synthesis of (R)-4-((S)-1-fluoro-12-(((S,E)-4-(fluoromethylene)-1,3-dimethylpiperidin-3-yl)methoxy)-5a,6,9,10-tetrahydro-5H,8H-4,7-dioxa-3,10a,11,13-tetraazanaphtho[1,8-ab]hepten-2-yl)-5-methyl-5,6,7,8-tetrahydronaphthol

[0172] (S)-1-fluoro-12-(((S,E)-4-(fluoromethylene)-1,3-dimethylpiperidin-3-yl)methoxy)-2-((R)-3-(methoxymethoxy)-8-methyl-5,6,7,8-tetrahydronaphth-1-yl)-5a,6,9,10-tetrahydro-5H,8H-4,7-dioxa-3,10a,11,13-tetraazanaphtho[1,8-ab]hepten (35 mg, 0.05 mmol) was dissolved in acetonitrile (1 mL), and then a 1,4-dioxane solution (0.5 mL) in 4N HCl was added. The reaction was stirred at room temperature for 30 minutes. The reaction was quenched with saturated sodium bicarbonate aqueous solution, extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and purified by TLC to give a white solid (24 mg, yield: 73%). MS:(ESI)m / z=608.46[M+H] + .

[0173] Examples 147-204 were obtained by applying the preparation method of Example 146.

[0174] Example 205: Synthesis of (2,2-dimethyl-1,3-dioxolane-4-yl)methyl((R)-4-((5S,5aS,6R,9S)-1-fluoro-12-((S,E)-4-(fluoromethylene)-1,3-dimethylpiperidin-3-yl)methoxy)-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaza-6,9-methylnaphtho[1,8-ab]hept-2-yl)-5-methyl ester-5,6,7,8-tetrahydronaphtho-2-yl) carbonate

[0175] Step 1: Synthesis of tert-butyl(5S,5aS,6R,9S)-1-fluoro-12-((S,E)-4-(fluoromethylene)-1,3-dimethylpiperidin-3-yl)methoxy)-2-((R)-3-hydroxy-8-methyl-5,6,7,8-tetrahydronaphth-1-yl)-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaza-6,9-methylnaphtho[1,8-ab]heptane-14 carboxylic acid ester

[0176] At room temperature, a methanol solution (2 mL) of di-tert-butyl dicarbonate (21.8 mg, 0.1 mmol) was added dropwise to a methanol solution (3 mL) of (R)-4-((5S,5aS,6R,9S)-1-fluoro-12-((S,E)-4-(fluoromethylene)-1,3-dimethylpiperidin-3-yl)methoxy)-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaza-6,9-methylnaphtho[1,8-ab]hept-2-yl)-5-ethyl-5,6,7,8-tetrahydronaphthyl-2-phenol (64 mg, 0.1 mmol). The reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was concentrated under reduced pressure, and the residue was dissolved in ethyl acetate (5 mL). Concentrating this solution under reduced pressure yielded a yellow solid (74 mg, 0.1 mmol, crude yield 100%). The crude product was used directly in the next step. MS m / z: 733.6 [M+H]+.

[0177] Step 2: Synthesis of tert-butyl(5S,5aS,6R,9S)-2-((8R)-3-((((2,2-dimethyl-1,3-dioxolane-4-yl)methoxy)carbonyl)oxy)-8-methyl-5,6,7,8-tetrahydronaphth-1-yl)-1-fluoro-12-((S,E)-4-(fluoromethylene)-1,3-dimethylpiperidin-3-yl)formyl)-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaza-6,9-methylnaphtho[1,8-ab]heptane-14-carboxylic acid ester

[0178] At room temperature, methyl 2,2-dimethyl-1,3-dioxane-3-yl)methoxy)-2-((R)-3-hydroxy-8-methyl-5,6,7,8-tetrahydronaphthyl-1-yl)-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaza-6,9-methylnaphtho[1,8-ab]heptane-14-carboxylic acid ester (74 mg, 0.12 mmol) and triethylamine (28 μL) in acetonitrile (2 mL) was added to (45 mg, 0.2 mmol) of (2,2-dimethyl-1,3-dioxolane-4-yl)1H-imidazolium-1-carboxylate. The resulting solution was stirred overnight at room temperature. The solution was diluted with ethyl acetate (2 mL), concentrated by rotary evaporation to obtain a yellow solid, and then treated under high vacuum with an oil pump. Preparative TLC (ethyl acetate: petroleum ether = 2:1) was used to separate the solid to a pale yellow solid (60 mg, 0.067 mmol, yield 67%). MS m / z: 891.6 [M+H]+.

[0179] Step 3: Synthesis of (2,2-dimethyl-1,3-dioxolane-4-yl)methyl((R)-4-((5S,5aS,6R,9S)-1-fluoro-12-((S,E)-4-(fluoromethylene)-1,3-dimethylpiperidin-3-yl)methoxy)-5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaza-6,9-methylnaphtho[1,8-ab]hept-2-yl)-5-methyl ester-5,6,7,8-tetrahydronaphtho-2-yl) carbonate

[0180] Under ice-water bath cooling, the tert-butyl(5S,5aS,6R,9S)-2-((8R)-3-((((2,2-dimethyl-1,3-dioxolane-4-yl)methoxy)carbonyl)oxy)-8-methyl-5,6,7,8-tetrahydronaphthyl)-1-fluoro-12-((S,E)-4-(fluoromethylene)-1,3-dimethylpiperidin-3-yl)formyl Trimethylsilyl trifluoromethanesulfonate (75 mg, 0.34 mmol) was added dropwise to an anhydrous dichloromethane (2 mL) solution of 5-methyl-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaza-6,9-methylnaphtho[1,8-ab]heptane-14-carboxylic acid ester (60 mg, 0.067 mmol). The resulting solution was stirred at this temperature for 4 hours. The reaction mixture was added dropwise to a saturated aqueous solution of sodium bicarbonate (10 mL) cooled in an ice-water bath and stirred vigorously. The mixture was extracted with dichloromethane (10 mL x 3), the organic phases were combined, washed with saturated brine, and dried over sodium sulfate. The solution was concentrated under reduced pressure to give a yellow viscous substance, which was purified by preparative TLC (methanol:dichloromethane = 1:8) to give an off-white solid (2 mg, 0.0025 mmol, yield 4%). MS m / z:791.5[M+H]+.

[0181] Examples 206-219 were obtained by applying the preparation method of Example 205.

[0182] Example 219: Synthesis of (S)-4-(8-fluoro-2-((S,Z)-2-(fluoromethylene)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy-d2)-7-((R)-3-hydroxy-8-methyl-5,6,7,8-tetrahydronaphth-1-yl)pyrido[4,3-d]pyrimidin-4-yl)-6-methyl-1,4-oxazin-6-ol

[0183] Step 1: (S)-4-(7-chloro-8-fluoro-2-(((S,E)-4-(fluoromethylene)-1,3-dimethylpiperidin-3-yl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)-6-methyl-1,4-oxazacycloheptan-6-ol

[0184] (S)-4-(2,7-dichloro-8-fluoropyridino[4,3-d]pyrimidin-4-yl)-6-methyl-1,4-oxazolidine-6-ol (22 mg, 0.0634 mmol) and (S,Z)-(2-(fluoromethylene)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methane-d2-ol (13.2 mg, 0.0761 mmol) were dissolved in anhydrous 1,4-dioxane (2 mL), followed by the addition of DIEA (11.2 mg, 0.0864 mmol). The mixture was heated to 95 °C and stirred for 24 hours. After the reaction was complete, the mixture was cooled to room temperature, diluted with water, extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and purified by TLC (DCM / MeOH = 20 / 1) to give a white solid (6 mg, yield: 20%). MS:(ESI)m / z=484.48[M+H] +

[0185] Step 2: (S)-4-(8-fluoro-2-((S,Z)-2-(fluoromethylene)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy-d2)-7-((R)-3-(methoxymethoxy)-8-methyl-5,6,7,8-tetrahydronaphthyl)pyrido[4,3-d]pyrimidin-4-yl)-6-methyl-1,4-oxazin-6-ol

[0186] The compound (S)-4-(7-chloro-8-fluoro-2-(((S,E)-4-(fluoromethylene)-1,3-dimethylpiperidin-3-yl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)-6-methyl-1,4-oxazacycloheptan-6-ol (6 mg, 0.0124 mmol), (R)-2-(3-(methoxymethoxy)-8-methyl-5,6,7,8-tetrahydronaphthyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborane (4.8 mg, 0.0149 mmol), potassium phosphate (7.6 mg, 0.036 mmol), and CataCXium A Pd G4 (1.4 mg, 0.00186 mmol) were purged three times with argon. A mixed solvent of dioxane (1.6 mL) and water (0.4 mL) was then added. The mixture was stirred overnight at 70°C. After cooling to room temperature, it was extracted with ethyl acetate (15 mL) and water (15 mL). The aqueous phase was extracted with ethyl acetate (10 mL) after separation. The combined organic phases were washed with saturated brine (10 mL) and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure, and the crude product was purified by preparative TLC (dichloromethane:methanol = 20:1) to give a white solid (6 mg, 0.00917 mmol, yield 74%). MS: (ESI) m / z = 654.54 [M+H] +

[0187] Step 3: (S)-4-(8-fluoro-2-((S,Z)-2-(fluoromethylene)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy-d2)-7-((R)-3-hydroxy-8-methyl-5,6,7,8-tetrahydronaphth-1-yl)pyrido[4,3-d]pyrimidin-4-yl)-6-methyl-1,4-oxazin-6-ol

[0188] The compound (S)-4-(8-fluoro-2-((S,Z)-2-(fluoromethylene)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy-d2)-7-((R)-3-(methoxymethoxy)-8-methyl-5,6,7,8-tetrahydronaphthyl)pyrido[4,3-d]pyrimidin-4-yl)-6-methyl-1,4-oxazin-6-ol (6 mg, 0.00917 mmol) was dissolved in acetonitrile (1 mL), and then a 1,4-dioxane solution (0.5 mL) of 4N HCl was added. The mixture was stirred at room temperature for 20 minutes. After the reaction was complete, a saturated aqueous sodium bicarbonate solution was added to quench the reaction. The mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated to obtain a crude product. The crude product was purified by preparative TLC (DCM / MeOH = 15 / 1) to give a white solid (3 mg, 0.00492 mmol, yield 54%), MS: (ESI) m / z = 610.51 [M+H). + .

[0189] Examples 220-246 were obtained by applying the preparation method of Example 219.

[0190] Biological evaluation experiments:

[0191] Example 1: AGS cell viability assay

[0192] This experiment evaluated and verified the inhibitory activity of the compounds contained in this invention on the proliferation of gastric cancer cell line AGS carrying the KRAS G12D mutation.

[0193] Cell information:

[0194] Main experimental materials, reagents and instruments:

[0195] 1. Cells: AGS for gastric cancer was purchased from Shanghai Enzyme Research Biotechnology Co., Ltd.

[0196] 2. Reagents: F12K medium (Gibco), fetal bovine serum (enzyme-based), Tryple cell digestion solution (Gibco), MTT (Solepro), DMSO (Solepro), DPBS (Solepro).

[0197] 3. Instruments: 37℃, 5% CO2 incubator (Likang), microplate reader (UTRAO), biosafety cabinet (Likang), cell counting chamber, microscope (Aote Optics).

[0198] 4. Experimental consumables: 96-well cell culture plate (Corning, 3599), 96-well round-bottom refill plate (Corning, 3799).

[0199] Cell viability testing experimental steps:

[0200] 1. Digest cells in the logarithmic growth phase, terminate digestion with fresh culture medium, and count viable cells using the trypan blue exclusion method to ensure cell viability is above 95%. Adjust the cell concentration to an appropriate density with fresh culture medium, add 90 μL of cell suspension to each well (1000 cells per well), and fill the other edge wells with sterile deionized water.

[0201] 2. Incubate overnight at 37°C in a 5% CO2 incubator.

[0202] 3. Experimental drug preparation: Dissolve the drug in DMSO to prepare a 20mM stock solution, then dilute it with DMSO to prepare a 1mM solution. Serially dilute this solution nine times at a 1:3 ratio to obtain 1000-fold compound gradient solutions. Add 2 μL of each gradient compound solution to 198 μL of cell culture medium to obtain a 10× gradient compound solution. Add 10 μL of the 10× compound solution to a 96-well culture plate inoculated with cells, with three replicates for each drug concentration. The control group was prepared with culture medium containing 0.1% DMSO. After the procedure, place the cell culture plate in an incubator and incubate for 72 hours.

[0203] 4. After incubation, add 10 μL of MTT solution (5 mg / mL) to each well and continue incubation for 4 hours.

[0204] 5. Carefully aspirate the culture medium from the wells, add 150 μL DDMSO to each well, and shake rapidly for 10 seconds in a microplate reader to fully dissolve the crystals. Measure the absorbance (OD) at a wavelength of 492 nm.

[0205] 6. Calculate the cell viability per well using the formula (Cell viability = (OD of test drug - OD of blank control) / (OD of solvent control - OD of blank control)). Analyze the viability data using GraphPad software to obtain the dose-response curve and calculate the IC50. 50 The values ​​and results are shown in the table below:

[0206] Example 2: GP2D cell viability assay

[0207] This experiment evaluated and verified the inhibitory activity of the compounds contained in this invention on the proliferation of the GP2D colon cancer cell line carrying the KRAS G12D mutation.

[0208] Cell information:

[0209] Main experimental materials, reagents and instruments:

[0210] 1. Cells: GP2D cells for colorectal cancer were purchased from Zhejiang Meisen Cell Technology Co., Ltd.

[0211] 2. Reagents: DMEM medium (Gibco), fetal bovine serum FBS (Enzyme Research), Tryple cell digestion solution (Gibco), MTT (Solepro), DMSO (Solepro), DPBS (Solepro).

[0212] 3. Instruments: 37℃, 5% CO2 incubator (Likang), microplate reader (UTRAO), biosafety cabinet (Likang), cell counting chamber, microscope (Aote Optics).

[0213] 4. Experimental consumables: 96-well cell culture plate (Corning, 3599), 96-well round-bottom refill plate (Corning, 3799).

[0214] Cell viability testing experimental steps:

[0215] 1. Digest cells in the logarithmic growth phase, terminate digestion with fresh culture medium, and count viable cells using the trypan blue exclusion method to ensure cell viability is above 95%. Adjust the cell concentration to an appropriate density with fresh culture medium, add 90 μL of cell suspension to each well (3000 cells per well), and fill the other edge wells with sterile deionized water.

[0216] 2. Incubate overnight at 37°C in a 5% CO2 incubator.

[0217] 3. Experimental drug preparation: Dissolve the drug in DMSO to prepare a 20mM stock solution, then dilute it with DMSO to prepare a 1mM solution. Serially dilute this solution nine times at a 1:3 ratio to obtain 1000-fold compound gradient solutions. Add 2 μL of each gradient compound solution to 198 μL of cell culture medium to obtain a 10× gradient compound solution. Add 10 μL of the 10× compound solution to a 96-well culture plate inoculated with cells, with three replicates for each drug concentration. The control group was prepared with culture medium containing 0.1% DMSO. After the procedure, place the cell culture plate in an incubator and incubate for 72 hours.

[0218] 4. After incubation, add 10 μL of MTT solution (5 mg / mL) to each well and continue incubation for 4 hours.

[0219] 5. Carefully aspirate the culture medium from the wells, add 150 μL DDMSO to each well, and shake rapidly for 10 seconds in a microplate reader to fully dissolve the crystals. Measure the absorbance (OD) at a wavelength of 492 nm.

[0220] 6. Calculate the cell viability per well using the formula (Cell viability = (OD of test drug - OD of blank control) / (OD of solvent control - OD of blank control)). Analyze the viability data using GraphPad software to obtain the dose-response curve and calculate the IC50. 50 The values ​​and results are shown in the table below:

[0221] Example 3: PANC-04-03 Cell Viability Assay

[0222] This experiment evaluated and verified the inhibitory activity of the compounds contained in this invention on the proliferation of pancreatic cancer cell line PANC-04-03 carrying the KRAS G12D mutation.

[0223] Cell information:

[0224] Main experimental materials, reagents and instruments:

[0225] 1. Cells: PANC-04-03 were purchased from Nanjing Kebai Biotechnology Co., Ltd.

[0226] 2. Reagents: RPMI 1640 medium (Gibco), fetal bovine serum (Enzyme Research), 100X ITS-G (ProBio), Tryple cell digestion solution (Gibco), MTT (Solepro), DMSO (Solepro), DPBS (Solepro).

[0227] 3. Instruments: 37℃, 5% CO2 incubator (Likang), microplate reader (UTRAO), biosafety cabinet (Likang), cell counting chamber, microscope (Aote Optics).

[0228] 4. Experimental consumables: 96-well cell culture plate (Corning, 3599), 96-well round-bottom refill plate (Corning, 3799).

[0229] Cell viability testing experimental steps:

[0230] 1. Digest cells in the logarithmic growth phase, terminate digestion with fresh culture medium, and count viable cells using the trypan blue exclusion method to ensure cell viability is above 95%. Adjust the cell concentration to an appropriate density with fresh culture medium, add 90 μL of cell suspension to each well (2000 cells per well), and fill the other edge wells with sterile deionized water.

[0231] 2. Incubate overnight at 37°C in a 5% CO2 incubator.

[0232] 3. Experimental drug preparation: Dissolve the drug in DMSO to prepare a 20mM stock solution, then dilute it with DMSO to prepare a 1mM solution. Serially dilute this solution nine times at a 1:3 ratio to obtain 1000-fold compound gradient solutions. Add 2 μL of each gradient compound solution to 198 μL of cell culture medium to obtain a 10× gradient compound solution. Add 10 μL of the 10× compound solution to a 96-well culture plate inoculated with cells, with three replicates for each drug concentration. The control group was prepared with culture medium containing 0.1% DMSO. After the procedure, place the cell culture plate in an incubator and incubate for 72 hours.

[0233] 4. After incubation, add 10 μL of MTT solution (5 mg / mL) to each well and continue incubation for 4 hours.

[0234] 5. Carefully aspirate the culture medium from the wells, add 150 μL DDMSO to each well, and shake rapidly for 10 seconds in a microplate reader to fully dissolve the crystals. Measure the absorbance (OD) at a wavelength of 492 nm.

[0235] 6. Calculate the cell viability per well using the formula (Cell viability = (OD of test drug - OD of blank control) / (OD of solvent control - OD of blank control)). Analyze the viability data using GraphPad software to obtain the dose-response curve and calculate the IC50. 50 The values ​​and results are shown in the table below:

[0236] Example 4: Pharmacokinetic characteristics of the compounds of the present invention in SD rats

[0237] The pharmacokinetic characteristics of the representative compounds of this invention were evaluated using pharmacokinetic experiments on SD rats.

[0238] Male SD rats, aged 6-8 weeks, were purchased from Spiford (Beijing) Biotechnology Co., Ltd.

[0239] Oral drug preparation: Weigh the compound from the examples into a 5% DMSO / 10% Solutol / 85% saline solution. First, add 5% DMSO according to the total volume ratio, vortex and sonicate until completely dissolved. Then add 10% Solutol, vortex and sonicate until completely dissolved. Finally, add 85% PBS, vortex and sonicate to obtain a clear solution with a concentration of 3 mg / mL. SD rats were administered the drug formulation by gavage at a dose of 10 mL / kg. Blood samples were collected from the jugular vein at 0.25 h, 0.5 h, 1.0 h, 2.0 h, 4.0 h, 6.0 h, 8.0 h, and 24.0 h post-administration. 50 μL of each sample was placed in an EDTA-K2 tube, centrifuged at low temperature for 6 minutes, and plasma was collected and stored at -80℃ for analysis. The final results were obtained using LC-MS / MS.

[0240] Pharmacokinetic parameters of the compounds of this invention in rats after oral administration:

[0241] NA: Not detected

[0242] The "positive compound 1" used in the rat pharmacokinetic evaluation of the compound in this application is Example 229 in WO2022247760A1, and its chemical structure is as follows:

[0243] Experimental results show that the compounds in the embodiments of this invention have good pharmacokinetic properties in rat pharmacokinetic evaluation.

[0244] Example 5: NCI-H441 cell viability assay

[0245] This experiment evaluated and verified the inhibitory activity of the compounds contained in this invention on the proliferation of pancreatic cancer cell line NCI-H441 carrying the KRAS G12V mutation.

[0246] Cell information:

[0247] Main experimental materials, reagents and instruments:

[0248] 1. Cells: NCI-H441 were purchased from Shanghai Enzyme Research Biotechnology Co., Ltd.

[0249] 2. Reagents: RPMI 1640 medium (Gibco), fetal bovine serum (enzyme-based), Tryple cell digestion solution (Gibco), MTT (Solepro), DMSO (Solepro), DPBS (Solepro).

[0250] 3. Instruments: 37℃, 5% CO2 incubator (Likang), microplate reader (UTRAO), biosafety cabinet (Likang), cell counting chamber, microscope (Aote Optics).

[0251] 4. Experimental consumables: 96-well cell culture plate (Corning, 3599), 96-well round-bottom refill plate (Corning, 3799).

[0252] Cell viability testing experimental steps:

[0253] 1. Digest cells in the logarithmic growth phase, terminate digestion with fresh culture medium, and count viable cells using the trypan blue exclusion method to ensure cell viability is above 95%. Adjust the cell concentration to an appropriate density with fresh culture medium, add 90 μL of cell suspension to each well (2000 cells per well), and fill the other edge wells with sterile deionized water.

[0254] 2. Incubate overnight at 37°C in a 5% CO2 incubator.

[0255] 3. Experimental drug preparation: Dissolve the drug in DMSO to prepare a 20mM stock solution, then dilute it with DMSO to prepare a 1mM solution. Serially dilute this solution nine times at a 1:3 ratio to obtain 1000-fold compound gradient solutions. Add 2 μL of each gradient compound solution to 198 μL of cell culture medium to obtain a 10× gradient compound solution. Add 10 μL of the 10× compound solution to a 96-well culture plate inoculated with cells, with three replicates for each drug concentration. The control group was prepared with culture medium containing 0.1% DMSO. After the procedure, place the cell culture plate in an incubator and incubate for 72 hours.

[0256] 4. After incubation, add 10 μL of MTT solution (5 mg / mL) to each well and continue incubation for 4 hours.

[0257] 5. Carefully aspirate the culture medium from the wells, add 150 μL DDMSO to each well, and shake rapidly for 10 seconds in a microplate reader to fully dissolve the crystals. Measure the absorbance (OD) at a wavelength of 492 nm.

[0258] 6. Calculate the cell viability per well using the formula (Cell viability = (OD of test drug - OD of blank control) / (OD of solvent control - OD of blank control)). Analyze the viability data using GraphPad software to obtain the dose-response curve and calculate the IC50. 50 value.

[0259] Example 6: SW620, H358, and A549 cell viability assay

[0260] This experiment evaluated and verified the inhibitory activity of the compounds contained in this invention on the proliferation of colon cancer cell line SW620 carrying the KRAS G12V mutation.

[0261] Cell information:

[0262] Main experimental materials, reagents and instruments:

[0263] 1. Cells: SW620 and H358 were purchased from Shanghai Enzyme Research Biotechnology Co., Ltd., and A549 was donated to Tianjin Medical University.

[0264] 2. Reagents: L-15 medium (source culture), 1640 medium (Gibco), F12K medium (Gibco), fetal bovine serum (enzyme-based), Tryple cell digestion solution (Gibco), MTT (Solepro), DMSO (Solepro), DPBS (Solepro).

[0265] 3. Instruments: 37℃, 5% CO2 incubator (Likang), microplate reader (UTRAO), biosafety cabinet (Likang), cell counting chamber, microscope (Aote Optics).

[0266] 4. Experimental consumables: 96-well cell culture plate (Corning, 3599), 96-well round-bottom refill plate (Corning, 3799).

[0267] Cell viability testing experimental steps:

[0268] 1. Digest cells in the logarithmic growth phase, terminate digestion with fresh culture medium, and count viable cells using the trypan blue exclusion method to ensure cell viability is above 95%. Adjust the cell concentration to an appropriate density with fresh culture medium, add 90 μL of cell suspension to each well, with 1000 to 5000 cells per well, and fill the other edge wells with sterile deionized water.

[0269] 2. Incubate overnight at 37°C in a 5% CO2 incubator.

[0270] 3. Experimental drug preparation: Dissolve the drug in DMSO to prepare a 20mM stock solution, then dilute it with DMSO to prepare a 1mM solution. Serially dilute this solution nine times at a 1:3 ratio to obtain 1000-fold compound gradient solutions. Add 2 μL of each gradient compound solution to 198 μL of cell culture medium to obtain a 10× gradient compound solution. Add 10 μL of the 10× compound solution to a 96-well culture plate inoculated with cells, with three replicates for each drug concentration. The control group was prepared with culture medium containing 0.1% DMSO. After the procedure, place the cell culture plate in an incubator and incubate for 72 hours.

[0271] 4. After incubation, add 10 μL of MTT solution (5 mg / mL) to each well and continue incubation for 4 hours.

[0272] 5. Carefully aspirate the culture medium from the wells, add 150 μL DDMSO to each well, and shake rapidly for 10 seconds in a microplate reader to fully dissolve the crystals. Measure the absorbance (OD) at a wavelength of 492 nm.

[0273] 6. Calculate the cell viability per well using the formula (Cell viability = (OD of test drug - OD of blank control) / (OD of solvent control - OD of blank control)). Analyze the viability data using GraphPad software to obtain the dose-response curve and calculate the IC50. 50 value.

[0274] The test results for some compounds are shown in the table below:

Claims

1. A compound of formula (I) or a pharmaceutically acceptable salt, solvate, tautomer, stereoisomer, or prodrug thereof, wherein the compound of formula (I) is: in: Y is It is a 4- to 12-element saturated or partially saturated single ring, bridged ring, or helical ring, wherein the saturated or partially saturated single ring, bridged ring, or helical ring is optionally composed of one or more R 1 Replace, X 1 Selected from N and CR 4 ;X 2 Selected from NR 4 , O, or C(R) 4 )2, R 1 Selected from hydrogen, halogens, optional halogenated elements, hydroxyl groups, C 1-6 Alkoxy, deuterated C 1-6 Alkoxy, -NR 1a R 1b C replaced by deuterium 1- 6-alkyl, CN, -OR 1a -SR 1a -NR 1a R 1b -S(O)R 1a -S(O)2R 1a -CH2OR 1a -C(O)OR 1a -NR 1a C(O)R 1b -C(O)NR 1a R 1b -S(O)2N(R) 1a R 1b )2 and 5- to 6-membered heteroaryl, wherein R 1a and R 1b Each independently is hydrogen, C 1-6 Alkyl, hydroxyl C 1-6 Alkyl, halogenated or deuterated C 1-6 Alkyl and C 1-6 Alkoxy C 1-6 Alkyl-, C 3-6 cycloalkyl; R 4 Selected from hydrogen, halogens, and C atoms optionally substituted with halogens or hydroxyl groups. 1-6 Alkyl, CN, -OR 4a -SR 4a -S(O)R 4a -S(O)2R 4a -C(O)R 4a -C(O)OR 4a -NR 4a C(O)R 4b -C(O)NR 4a R 4b and -S(O)2N(R 4a R 4b )2, where R 4a and R 4b Each is independently hydrogen, a 4-6 membered oxecyclic alkyl group optionally substituted with methyl, dimethyl or isopropyl, C 1-6 Alkyl, hydroxyl C 1-6 Alkyl, Halogenated C 1- 6-alkyl and C 1-6 Alkoxy C 1-6 alkyl-; L represents a single bond, -O-, -S-, or -NR. La -、-O-(CR La R Lb ) t -、-S-(CR La R Lb ) t -、-NR c -(CR La R Lb ) t -、-(CR La R Lb ) t -O-、-(CR La R Lb ) t -S-、-(CR La R Lb ) t -NR Lc -, -C(O)-, -SO2-, -SO-, -C(O)-O-, -OC(O)-, -C(O)-NR Lc -or-N Lc C(O)-, where R La R Lb and R Lc Each is independently selected from hydrogen, deuterium, and C. 1-6 Alkyl group, or R group attached to the same carbon atom La and R Lb Together with the attached carbon atom, it forms a C3-C6 cycloalkyl group, wherein t is an integer from 1 to 6; R 2 C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 3-8 Cycloalkyl, aryl, heteroaryl, or heterocyclic, wherein the C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 3-8 Cycloalkyl, aryl, heteroaryl, and heterocyclic groups are each independently unsubstituted or converted by deuterium, halogen, cyano, or C. 1-6 Alkyl, Halogenated C 1-6 Alkyl, C 1-6 Alkylene, Halogenated C 1-6 Alkylene, hydroxyl C 1-6 Alkyl, C 1-6 Alkoxy C 1-6 Alkyl, oxo, -OR 2a -C(O)R 2a 、-(CR 2a R 2b ) m -OC(O)NR 2c R 2d -CO2R 2a -CONR 2c R 2d -NR 2c R 2d C 3-8 cycloalkyl, C 3-8 cycloalkyl C 1-6 One or more substitutions of alkyl, aryl, heteroaryl, and heterocyclic groups, wherein R 2a R 2b R 2c and R 2d Each is independently hydrogen, C 1-6 Alkyl, hydroxyl C 1-6 Alkyl, Halogenated C 1-6 Alkyl and C 1-6 Alkoxy C 1-6 Alkyl group, or R atom attached to the same nitrogen atom 2c and R 2d Together with the attached nitrogen atom, it forms a 4- to 6-membered heterocycle, the heterocycle containing 0, 1 or 2 additional heteroatoms selected from nitrogen, oxygen or sulfur as ring members, and wherein m is an integer from 1 to 6; R 3 It is an aryl or heteroaryl group, wherein the aryl or heteroaryl group is optionally composed of one or more R groups. 8 Replace; each R 8 Independently selected C groups are chosen from halogens, cyano groups, oxo groups, and C groups optionally substituted with halogens, cyano groups, hydroxyl groups, and deuterated groups. 1-6 Alkyl, C2-C6 alkenyl group optionally substituted with hydroxyl or deuterium, C2-C6 ynyl group optionally substituted with hydroxyl or deuterium, -OR 8a -SR 8a -S(O)2R 8a -P(=O)R 8a R 8b -NR 8a R 8b -C(O)NR 8a R 8b Optional halogenated or C 1-6 Alkyl-substituted C3-C6 cycloalkyl, C3-C8 cycloalkyl, heterocyclic, heteroaryl, and aryl groups, wherein R 8a and R 8b Each independent hydrogen atom, and C atoms that can be arbitrarily substituted with halogens. 1-6 Alkyl and C 1-6 Alkoxy-C 1-6 alkyl-; Q 1 Q 2 and Q 3 Each is independently N or CR 6 M 1 and M 2 Each is independently N or CR 7 The condition is Q 1 and M 1 At least one of them is N; Where R 6 and R 7 Each is independently hydrogen, halogen, hydroxyl, cyano, nitro, C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 3- 8-Cycloalkyl, aryl, heteroaryl or heterocyclic, -OR 6a -C(O)R 6a -CO2R 6a -CONR 6a R 6b or -NR 6a R 6b Wherein C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 3-8 Cycloalkyl, aryl, heteroaryl, and heterocyclic groups are each independently converted by oxidizing, halogenating, hydroxyling, or C-oxidizing. 1-4 Alkoxy, C 1-4 Alkyl, C 3-6 Cycloalkyl, nitro, cyano and -NR d R e One or more substitutions in, where R 6a R 6b R 6c and R 6d Each is independently hydrogen, C 3-6 Alkyl, C 1-6 Alkyl, hydroxyl C 1-6 Alkyl, Halogenated C 1-6 Alkyl and C 1-6 Alkoxy C 1-6 alkyl-; Alternatively, a substituent on Y and Q 3 The substituents on them are linked together to form a macrocyclic structure.

2. The compound of claim 1, wherein X 1 Let N, X 2 For NR 4 ,as well as for Where n is an integer between 2 and 4, and R 1 It is hydrogen, hydroxyl, or optionally halogenated, hydroxyl, C 1-6 Alkoxy, deuterated C 1-6 Alkoxy, -NR 1a R 1b C replaced by deuterium 1-6 Alkyl; preferably Y is Where R 1 For hydrogen, hydroxyl, C 1-6 Alkyl, Halogenated C 1-6 Alkyl, deuterated C 1- 6-alkyl, C 1-6 Alkoxy C 1-6 Alkyl- or deuterated C 1-6 Alkoxy C 1-6 Alkyl group; more preferably, Y is...

3. The compound of claim 1 or 2, wherein L is -O- or -O-(CH2). t -or Where t is an integer from 1 to 6, t1+t2 is an integer from 0 to 5, and t3 is an integer from 1 to 3; preferably L is -O-CH2- or -O-.

4. The compound of any one of claims 1-3, wherein Q 1 and M 1 All are N.

5. The compound of any one of claims 1-4, wherein Q 3 and M 2 All are CH.

6. The compound of any one of claims 1-5, wherein Q 2 Let N be the number of elements in the array.

7. The compound of any one of claims 1-6, wherein R 2 The heterocyclic group is unsubstituted or modified by deuterium, halogen, or C. 1-6 Alkyl, C 1-6 Alkylene, Halogenated C 1-6 Alkylene, -OR 2a and -(CR 2a R 2b ) m -OC(O)NR 2c R 2d One or more substitutions in the equation, wherein each variable is defined as in equation (I); Preferably, the heterocyclic group is unsubstituted or replaced by deuterium, halogen, or C. 1-6 Alkyl, C 1-6 Alkylene, Halogenated C 1-6 Alkylene and -OR 2a One or more of the heterocyclic group is substituted; more preferably, the heterocyclic group is unsubstituted or substituted by one or two of halogen, methyl, methoxy, methylene, and halomethylene. Preferably R 2 It is a 4- to 8-membered monocyclic heterocycle containing 1, 2, or 3 heteroatoms selected from oxygen, nitrogen, and sulfur as ring members, or a 6- to 12-membered bicyclic heterocycle (preferably a bridged bicyclic) containing 1, 2, or 3 heteroatoms selected from oxygen, nitrogen, and sulfur as ring members, wherein the monocyclic or bicyclic heterocyclic group is unsubstituted or substituted with deuterium, halogen, or C. 1-6 Alkyl, C 1-6 Alkylene, Halogenated C 1-6 Alkylene, -OR 2a and -(CR 2a R 2b ) m -OC(O)NR 2c R 2d One or more substitutions in the heterocyclic group, wherein each variable is defined as in equation (I); preferably, the heterocyclic group is unsubstituted or halogenated, C 1-6 Alkyl, C 1-6 Alkylene, Halogenated C 1-6 Alkylene and -OR 2a One or more substitutions in; More preferably, the heterocyclic group is unsubstituted or substituted by one or two of deuterium, halogen, methyl, methoxy, methylene, and halomethylene; Preferably R 2 It is a monocyclic heterocyclic ring, which is piperidinyl, aza-butyl, or pyrrolidinyl, said ring being unsubstituted or converted by one or two deuterium, halogen, or C. 1-6 Alkyl, C 1-6 Alkylene or halogenated C 1-6 Alkyl substituent; Preferably R 2 It is a bicyclic heterocycle, namely octahydrocyclopentadiene, wherein at least one carbon atom is replaced by a nitrogen atom, and one of the other carbon atoms is optionally replaced by an oxygen atom.

8. The compound of any one of claims 1-7, wherein L is -O-CH2- or and R 2 The heterocyclic group is unsubstituted or modified by deuterium, halogen, or C. 1-6 Alkyl, C 1-6 Alkylene, Halogenated C 1-6 Alkylene, -OR 2a and -(CR 2a R 2b ) m -OC(O)NR 2c R 2d One or more substitutions in the equation, wherein each variable is defined as in equation (I); Preferably, the heterocyclic group is unsubstituted or replaced by deuterium, halogen, or C. 1-6 Alkyl, C 1-6 Alkylene, Halogenated C 1-6 Alkylene and -OR 2a One or more substitutions in; More preferably, the heterocyclic group is unsubstituted or substituted by one or two of deuterium, halogen, methyl, methoxy, methylene or halomethylene; Preferably, L is -O-CH2- or and R 2 It is a 4- to 8-membered monocyclic heterocycle containing 1, 2, or 3 heteroatoms selected from oxygen, nitrogen, and sulfur as ring members, or a 6- to 12-membered bicyclic heterocycle (preferably a bridged bicyclic) containing 1, 2, or 3 heteroatoms selected from oxygen, nitrogen, and sulfur as ring members, wherein the monocyclic or bicyclic heterocyclic group is unsubstituted or converted by halogen, C 1-6 Alkyl, C 1-6 Alkylene, Halogenated C 1-6 Alkylene, -OR 2a and -(CR 2a R 2b ) m -OC(O)NR 2c R 2d One or more substitutions in the equation, wherein each variable is defined as in equation (I); Preferably, the heterocyclic group is unsubstituted or replaced by deuterium, halogen, or C. 1-6 Alkyl, C 1-6 Alkylene, Halogenated C 1-6 Alkylene and -OR 2a One or more substitutions in; More preferably, the heterocyclic group is unsubstituted or substituted with one or two of deuterium, halogen, methyl, methoxy, methylene and halomethylene; Preferably, L is -O-CH2- or and R 2 It is a monocyclic heterocyclic ring, which is piperidinyl, aza-butyl, or pyrrolidinyl, wherein the ring is unsubstituted or converted by one or two halogens, C 1-6 Alkyl, C 1-6 Alkylene, Halogenated C 1-6 Alkyl substituent; Preferably, L is -O-CH2- or and R 2 It is a bicyclic heterocycle, namely octahydrocyclopentadiene, wherein at least one carbon atom is replaced by a nitrogen atom, and one of the other carbon atoms is optionally replaced by an oxygen atom.

9. The compound of any one of claims 1-8, wherein LR 2 for 10. The compound of any one of claims 1-9, wherein R 3 It is an aryl or heteroaryl group of a benzo5-8 membered saturated ring or heterocycle, wherein the benzo5-8 membered saturated ring, aryl or heteroaryl group is substituented by 1 to 3 R groups. 8 Replace, R 8 Each element is independently selected from: hydrogen, halogens, or C elements optionally substituted with halogens. 1-6 Alkyl, C2-C6 ynyl, -P(=O)R 8a R 8b -NR 8a R 8b -OR 8a -SR 8a Optional halogenated or C 1-6 Alkyl-substituted C3-C6 cycloalkyl, wherein R 8a and R 8b Each independent hydrogen, -C(O)OC 1-6 Alkyl groups and -C(O)NHC 1-6 alkyl; Preferably, R 3 The benzo5-8 membered saturated ring is composed of 1 to 3 substituents R. 8 Replace, R 8 Each element is independently selected from: hydrogen, halogens, or C elements optionally substituted with halogens. 1-6 alkyl.

11. The compound of any one of claims 1-10, wherein R 3 for ,in: R 8 Preferred from -OH, -NH2, -OC(O)-C 1-6 Alkyl group, -OC(O)CH2CH3, -OC(O)NH-C 1-6 Alkyl or alkoxy.

12. The compound of claim 1, wherein the compound of formula (I) is as shown in formula (I-1): in Y is R 6 It is hydrogen, ethynyl, propynyl, methoxy, isopropoxy, or halogenated C. 1-6 Alkyl and C 1-6 Alkoxy; R 7 It is hydrogen, halogen, methoxy, -OCD3, halogenated or unsubstituted C 1-6 Alkoxy; LR 2 for The remaining variables are as defined in equation (I).

13. The compound of claim 1, wherein the compound of formula (I) is as shown in formula (I-1-1): in, R 1 It can be hydrogen, methyl, ethyl, -CH2-OCD3, CD3, or CD2CD3. R 6 It can be hydrogen, ethynyl, propynyl, methoxy, -OCD3, halogenated or unsubstituted C 1-6 Alkoxy; R 7 Preferably, it contains hydrogen, halogen, methoxy, -OCD3, halogenated or unsubstituted C. 1-6 Alkoxy; LR 2 for R 3 for The remaining variables are as defined in equation (I).

14. The compound of claim 1, wherein Q in the compound of formula (I) 3 For CR 6 And R 6 With R 1 The links form 7-8 elemental heterocyclic rings, as shown in equation (I-2): in: G 1 and G 2 Each of these is independent: single bond, -O-, -S-, -NR La -、-O-(CR La R Lb ) t -、-S-(CR La R Lb ) t -、-NR c -(CR La R Lb ) t -、-(CR La R Lb ) t -O-、-(CR La R Lb ) t -S-、-(CR La R Lb ) t -NR Lc -, -C(O)-, -SO2-, -SO-, -C(O)-O-, -OC(O)-, -C(O)-NR Lc -or-N Lc C(O)-, where R La R Lb and R Lc Each is independently selected from hydrogen and deuterated C. 1-6 Alkyl or C 1- 6 alkyl groups, and t is an integer from 1 to 6, with the remaining variables as defined for formula (I).

15. The compound of claim 1, wherein the compound of formula (I) is as shown in formula (I-2-1): in: G 1 and G 2 Each of these is independent: single bond, -O-, -S-, -NR La -、-O-(CR La R Lb ) t -、-S-(CR La R Lb ) t -、-NR c -(CR La R Lb ) t -、-(CR La R Lb ) t -O-、-(CR La R Lb ) t -S-、-(CR La R Lb ) t -NR Lc -, -C(O)-, -SO2-, -SO-, -C(O)-O-, -OC(O)-, -C(O)-NR Lc -or-N Lc C(O)-, where R La R Lb and R Lc Each is independently selected from hydrogen and deuterated C. 1-6 Alkyl or C 1- 6 alkyl groups, wherein t is an integer from 1 to 6, R 1 Preferably, it is hydrogen, methyl, ethyl, -CH2-OCD3, CD3, and CD2CD3, Q 2 Preferably N or CR 6 The remaining variables are as defined in equation (I).

16. The compound of claim 1, wherein the compound of formula (I) is as shown in formula (I-2-1): in: G 1 -G 2 -O-(CR) La R Lb ) t -、-S-(CR La R Lb ) t -、-NR c -(CR La R Lb ) t -、-(CR La R Lb ) t -O-、-(CR La R Lb ) t -S- or -(CR) La R Lb ) t -NR Lc -, where R La R Lb and R Lc Each is independently selected from hydrogen and deuterated C. 1-6 Alkyl or C 1-6 Alkyl groups, and wherein t is an integer from 1 to 6, R 1 Preferably, it is hydrogen, methyl, ethyl, -CH2-OCD3, CD3, and CD2CD3, Q 2 Preferably N or CR 6 The remaining variables are as defined in equation (I). In some implementations, G 1 -G 2 -O-(CHR) La )-、-S-(CHR La )-、-NH-(CHR La )-、-(CHR La )-O-、-(CHR La -S- or -(CHR) La )-NH-, where R La Selected from hydrogen and deuterated C 1-6 Alkyl or C 1-6 Alkyl, R 1 The preferred components are hydrogen, methyl, ethyl, -CH2-OH, -CH2-OCD3, CD3, and CD2CD3, with the remaining variables as defined for formula (I).

17. The compound of claim 1, wherein the compound of formula (I) is as shown in formula (I-2-2): in: n is an integer from 1 to 6, G 1 and G 2 Each of these is independent: single bond, -O-, -S-, -NR La -、-O-(CR La R Lb ) t -、-S-(CR La R Lb ) t -、-NR c -(CR La R Lb ) t -、-(CR La R Lb ) t -O-、-(CR La R Lb ) t -S-、-(CR La R Lb ) t -NR Lc -, -C(O)-, -SO2-, -SO-, -C(O)-O-, -OC(O)-, -C(O)-NR Lc -or-N Lc C(O)-, where R La R Lb and R Lc Each is independently selected from hydrogen and deuterated C. 1-6 Alkyl or C 1-6 Alkyl groups, and wherein t is an integer from 1 to 6, R 1 Preferably, the constituents are hydrogen, methyl, ethyl, -CH2-OH, -CH2-OCD3, CD3, and CD2CD3, Q. 2 Preferably N or CR 6 The remaining variables are as defined in equation (I).

18. The compound of claim 1, wherein the compound of formula (I) is as shown in formula (I-2-3): in: n is an integer from 1 to 6, G 1 and G 2 Each of these is independent: single bond, -O-, -S-, -NR La -、-O-(CR La R Lb ) t -、-S-(CR La R Lb ) t -、-NR c -(CR La R Lb ) t -、-(CR La R Lb ) t -O-、-(CR La R Lb ) t -S-、-(CR La R Lb ) t -NR Lc -, -C(O)-, -SO2-, -SO-, -C(O)-O-, -OC(O)-, -C(O)-NR Lc -or-N Lc C(O)-, where R La R Lb and R Lc Each is independently selected from hydrogen and deuterated C. 1-6 Alkyl or C 1-6 Alkyl groups, and wherein t is an integer from 1 to 6, R 1 Preferably, hydrogen, methyl, ethyl, trifluoromethyl, trifluoroethyl, C 3-6 Cycloalkyl, -CH2-OCD3, CD3 and CD2CD3, Q 2 Preferably N or CR 6 The remaining variables are as defined in equation (I).

19. The compound of claim 1, wherein the compound is a specific compound disclosed in this invention.

20. A pharmaceutical composition comprising the compound as described in any one of claims 1-19 or a pharmaceutically acceptable salt, tautomer, prodrug, or stereoisomer thereof.

21. Use of a compound as described in any one of claims 1-19, or a pharmaceutically acceptable salt, tautomer, prodrug, or stereoisomer thereof, in the preparation of a treatment for cancers associated with H-ras, K-ras, or N-ras inhibition.

22. The use as described in claim 21, wherein the cancer is a disease mediated by KRas mutation and KRAS amplification selected from pancreatic cancer, colon cancer, rectal cancer, lung adenocarcinoma, lung cancer, bile duct cancer, endometrial cancer, ovarian cancer, and leukemia; most preferably selected from pancreatic cancer, colon cancer, rectal cancer, lung adenocarcinoma, and bile duct cancer. Cancers mediated by G12C, G12D, G12V, G12A, G12R, G12S, G13D, and Q61H mutations.

23. The use according to claim 22, wherein the cancer is lung cancer, colorectal cancer, or pancreatic cancer.

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