WO2021027911A1 - Novel spirocyclic k-ras g12c inhibitor - Google Patents

Abstract

The present invention relates to a spirocyclic compound and a preparation method therefor and use thereof, in particular to a compound represented by formula (1) and a preparation method therefor, and use of the compound of formula (1) and optical isomers, crystal forms, and pharmaceutically acceptable salts thereof as irreversible inhibitors of a G12C mutant K-Ras protein in the preparation of drugs for Ras-related diseases such as anti-tumor.

Description

New spirocyclic K-Ras G12C inhibitor

This application claims the priority of the Chinese patent application CN2019107569656 whose filing date is August 15, 2019. This application quotes the full text of the aforementioned Chinese patent application.

Technical field

The present invention belongs to the field of medicinal chemistry, and more specifically, to a new type of K-Ras G12C inhibitor, its preparation method and the use method of the compound.

Background technique

The Ras protein family is an important signal transduction and transmission molecule in the cell, which plays an important role in growth and development. The analysis and research of a large number of in vitro tumor cells, animal models and human tumor samples show that the excessive activation of Ras family proteins is an early event in the development of human tumors and an important cause of the occurrence and development of a variety of cancers. Therefore, targeting and inhibiting the activity of Ras protein are important means to treat related tumors.

There are two forms of Ras protein. It binds to GDP and is in an inactive resting state. When cells receive signals such as growth factor stimulation, Ras protein binds to GTP and is activated. The activated Ras protein recruits a variety of signal transfer proteins, promotes the phosphorylation of downstream signal molecules such as ERK and S6, thereby activating the Ras signal transduction pathway, regulating cell growth, survival, migration and differentiation. The GTPase activity of Ras protein itself can hydrolyze GTP back to GDP. In addition, the presence of GTPase activating proteins (GAPs) in cells interacting with Ras greatly promotes the activity of Ras GTPase, thereby preventing excessive activation of Ras protein.

Mutations on K-Ras, H-Ras and N-Ras proteins in the Ras protein family are one of the common gene mutations in a variety of tumors, and are the main factors leading to the excessive activation of Ras protein in tumors. Compared with the wild-type Ras protein, these mutations cause the Ras protein activity to be unregulated, stably bind to GTP, and continue to activate, thereby promoting the growth, migration and differentiation of tumor cells. Among them, K-Ras protein mutations are the most common, accounting for 85% of all Ras mutations, while N-Ras (12%) and H-Ras (3%) are relatively rare. K-Ras mutations are extremely common in a variety of cancers: including pancreatic cancer (95%), colorectal cancer (45%) and lung cancer (25%), etc., and relatively rare in breast, ovarian, and brain cancers (< 2%). K-Ras mutation sites are mainly concentrated in G12, and G12C mutations are the most common. For example, in non-small cell lung cancer (NSCLC), K-Ras G12C accounts for 50% of all K-Ras mutations, followed by G12V and G12D. Genomics studies have shown that the K-Ras mutation in non-small cell lung cancer does not coexist with EGFR, ALK, ROS1, RET, and BRAF mutations, but coexists with mutations such as STK11, KEAP1, and TP53, suggesting that K-Ras mutations may be associated with STK11, KEAP1 Synergistic effect with TP53 mutation and other factors participates in the malignant transformation, proliferation and invasion of cells. In addition to tumors, the abnormal activation of Ras protein is also involved in non-neoplastic diseases such as diabetes and neurodegenerative diseases. It can be seen that small molecule compounds targeting Ras protein can enable a large number of cancer patients and Ras with specific gene mutations. Non-cancerous patients with excessive activation of the pathway benefit.

However, since the discovery of Ras mutations in tumors for 40 years, although we have a more in-depth understanding of the pathogenic mechanism of the Ras pathway, there is no effective target for a large number of patients with Ras protein mutations and excessive activation of the Ras pathway. The treatment of Ras protein. Therefore, the development of highly active small molecule inhibitors against Ras protein, especially K-Ras G12C protein with high mutation frequency, has important clinical significance.

Summary of the invention

The present invention aims to provide a class of compounds with general structural formula as shown in formula (1), or each of its optical isomers, crystal forms, pharmaceutically acceptable salts, hydrates or solvates:

In formula (1):

L is a chemical bond or NH;

A is a bivalent 4-12 membered saturated or partially saturated monocyclic, bicyclic, bridged or spiro ring containing 1-2 N atoms. The monocyclic, bicyclic, bridged or spiro ring may be optionally substituted by one R ² is substituted with one or more, when a plurality of R & lt substituted ^2, R ² may be the same or different, R ² is H, CN, C1-C3 alkyl, halogen substituted C1-C3 alkyl or cyano-substituted C1- C3 alkyl;

R ¹ is an aryl group or a heteroaryl group, and the aryl group or heteroaryl group can be substituted by 1-3 of the following groups: halogen, hydroxyl, amino, C1-C3 alkyl, C2-C4 alkenyl, C3 -C6 cycloalkyl, C1-C3 alkoxy, halogen-substituted C1-C3 alkyl or halogen-substituted C1-C3 alkoxy. When substituted by multiple substituents, the substituents may be the same or different;

Indicates that there is a single bond or double bond between V and Z, when Z is CO and V is

When, there is a single bond between V and Z; or, when V is N, and Z is

When, there is a double bond connection between V and Z;

R _a , R _b , R _c and R _d are all H, and R _e and R _f and their connected carbon atoms form a C3-C6 cycloalkyl or C4-C6 heterocyclic ring; or, R _c , R _d , R _e and R _f are both H, R _a and R _b and the carbon atom to which they are attached form a C3-C6 cycloalkyl or C4-C6 heterocyclic ring; _{or, R a, R b, R} e and R _f are both H, R _c and R _d and the carbon atom to which they are connected form a C3-C6 cycloalkyl or C4-C6 heterocyclic ring; and

E is an electrophilic moiety capable of forming a covalent bond with the cysteine residue at position 12 of the K-Ras mutant protein,

among them,

Y is a chemical bond or C1-C6 alkylene group;

R ³ is an amino-substituted alkyl group, cycloalkyl group, alkyl-substituted amido group, heterocyclic group, aryl group or heteroaryl group. The heterocyclic group, aryl group or heteroaryl group can be substituted by 1-3 The group is substituted: halogen, O, CN, OH, hydroxy substituted alkyl, dialkyl substituted amino, C1-C6 alkyl, C3-C6 cycloalkyl, halogen substituted C1-C3 alkyl or halogen substituted C1 -C3 alkoxy, when substituted by multiple substituents, the substituents may be the same or different.

In another preferred embodiment, the structure of the compound of general formula (1) is as shown in general formula (1A) or general formula (1B):

among them:

L is a chemical bond or NH;

A is a bivalent 4-12 membered saturated or partially saturated monocyclic, bicyclic, bridged or spiro ring containing 1-2 N atoms. The monocyclic, bicyclic, bridged or spiro ring may be optionally substituted by one R ² is substituted with one or more, when a plurality of R & lt substituted ^2, R ² may be the same or different, R ² is H, CN, C1-C3 alkyl, halogen substituted C1-C3 alkyl or cyano-substituted C1- C3 alkyl;

R ¹ is an aryl group or a heteroaryl group, and the aryl group or heteroaryl group can be substituted by 1-3 of the following groups: halogen, hydroxyl, amino, C1-C3 alkyl, C2-C4 alkenyl, C3 -C6 cycloalkyl, C1-C3 alkoxy, halogen-substituted C1-C3 alkyl or halogen-substituted C1-C3 alkoxy. When substituted by multiple substituents, the substituents may be the same or different;

Y is a chemical bond or C1-C6 alkylene group;

R ³ is an amino-substituted alkyl group, cycloalkyl group, alkyl-substituted amido group, heterocyclic group, aryl group or heteroaryl group. The heterocyclic group, aryl group or heteroaryl group can be substituted by 1-3 The group is substituted: halogen, O, CN, OH, hydroxy substituted alkyl, dialkyl substituted amino, C1-C6 alkyl, C3-C6 cycloalkyl, halogen substituted C1-C3 alkyl or halogen substituted C1 -C3 alkoxy, when it is substituted by multiple substituents, the substituents may be the same or different;

R _a , R _b , R _c and R _d are all H, and R _e and R _f and their connected carbon atoms form a C3-C6 cycloalkyl or C4-C6 heterocyclic ring; or, R _c , R _d , R _e and R _f are both H, R _a and R _b and the carbon atom to which they are attached form a C3-C6 cycloalkyl or C4-C6 heterocyclic ring; _{or, R a, R b, R} e and R _f are both H, R _c and R _d and the carbon atom to which they are connected form a C3-C6 cycloalkyl or C4-C6 heterocyclic ring; and

E is an electrophilic moiety capable of forming a covalent bond with the cysteine residue at position 12 of the K-Ras mutant protein.

In another preferred embodiment, in the general formula (1), general formula (1A) and general formula (1B), E is a group containing an electrophilic carbon-carbon double bond or a carbon-carbon triple bond.

In another preferred example, in the general formula (1), general formula (1A) and general formula (1B), E is:

Among them, R ⁴ is H, F, CF ₃ , OMe or -CH ₂ OMe, and R ⁵ is H, Me, Et, CN, -CONH ₂ , -CH ₂ F, -CHF ₂ , CF ₃ , -CH ₂ OH , CH ₂ OMe,

In another preferred example, in the general formula (1), general formula (1A) and general formula (1B), -ALE is:

Wherein, n is 1 or 2, L is a chemical bond or NH, and R ² is H, CN, C1-C3 alkyl, halogen substituted C1-C3 alkyl, or cyano substituted C1-C3 alkyl.

In another preferred example, in the general formula (1), general formula (1A) and general formula (1B), Y is a chemical bond, -CH ₂ -, -CH(Me)- or -CH ₂ CH ₂ -.

In another preferred example, in the general formula (1), general formula (1A) and general formula (1B), R ¹ is:

Wherein R ⁶ and R ^{7 are} independently H, halogen, hydroxy, amino, C1-C3 alkyl, C2-C4 alkenyl, C3-C6 cycloalkyl, C1-C3 alkoxy, halogen substituted C1-C3 alkyl Or halogen substituted C1-C3 alkoxy.

In another preferred embodiment, in the general formula (1), general formula (1A) and general formula (1B), R ³ is:

Where n is 1, 2 or 3, R ⁸ and R ^{9 are} independently H, halogen, hydroxyl, amino, C1-C3 alkyl, C2-C4 alkenyl, C3-C6 cycloalkyl, C1-C3 alkoxy , Halogen substituted C1-C3 alkyl or halogen substituted C1-C3 alkoxy, R ¹⁰ is C1-C3 alkyl, C3-C6 cycloalkyl, C3-C6 cycloalkyl alkyl, C1-C3 alkoxy alkane Group, halogen substituted C1-C3 alkyl, halogen substituted C3-C6 cycloalkyl or

In various embodiments, the compound has one of the structures listed in Table 1 below:

Table 1: List of representative compounds of the present invention:

Another object of the present invention is to provide a pharmaceutical composition, which contains a pharmacologically acceptable excipient or carrier, and the compound of the general formula (1) of the present invention, or each of its optical isomers, pharmaceutically acceptable The inorganic or organic salt as the active ingredient.

Another object of the present invention is to provide the use of the above-mentioned compound of the present invention, or each of its optical isomers, or pharmaceutically acceptable inorganic or organic salts, for preparing and treating RAS-related diseases.

It should be understood that the foregoing general description and the following detailed description of the present invention are both exemplary and illustrative, and are intended to provide a further description of the claimed invention.

Compound synthesis

The following specifically describes the preparation methods of the compounds of the general formula (1) of the present invention, but these specific methods do not constitute any limitation to the present invention.

The compound of general formula (1) described above can be synthesized using standard synthesis techniques or well-known techniques and methods combined in the text. In addition, the solvent, temperature and other reaction conditions mentioned here can be changed. The starting materials for the synthesis of the compound can be synthesized or obtained from commercial sources such as, but not limited to, Aldrich Chemical Co. (Milwaukee, Wis.) or Sigma Chemical Co. (St. Louis, Mo.). The compounds described herein and other related compounds with different substituents can be synthesized using well-known techniques and raw materials, including those found in March, ADVANCED ORGANIC CHEMISTRY 4 ^th Ed., (Wiley 1992); Carey and Sundberg, ADVANCED ORGANIC CHEMISTRY 4 ^th Ed., Vols. A and B (Plenum 2000, 2001), Green and Wuts, PROTECTIVE GROUPS IN ORGANIC SYNTHESIS 3 ^rd Ed., (Wiley 1999). The general method of compound preparation can be changed by using appropriate reagents and conditions for introducing different groups in the molecular formula provided herein.

In one aspect, the compounds described herein are according to methods well known in the art. However, the conditions of the method, such as reactants, solvents, bases, amounts of compounds used, reaction temperature, time required for the reaction, etc. are not limited to the following explanations. The compounds of the present invention can also be conveniently prepared by combining various synthetic methods described in this specification or known in the art, and such combinations can be easily performed by those skilled in the art to which the present invention belongs. On the one hand, the present invention also provides a method for preparing the compound represented by the general formula (1), which is prepared by the following general reaction scheme 1 or 2:

General reaction scheme 1

Embodiment of the compounds of formula (1) (Method A) was prepared according to the general reaction Scheme 1, wherein _{R a, R b, R c} , R d, R e, R f, R 1, R 2, R 3, A , E and Y are as defined above. As shown in the general reaction scheme 1, the raw material S (synthesized with reference to the method described in the preparation examples 1-9 of this patent) and the fragment A produce A1 under alkaline conditions, and the protective group of A1 (such as Boc) is removed to obtain A2, A2 and R1-X yielded A3, A3 a compound in the presence of an oxidant A4, A4 and reaction fragment R ³ -Y-OH to generate under appropriate conditions A5, A5 removing the protecting group (e.g., Cbz) to give A6, A6, and acid chlorides Or the acid anhydride compound reacts to form A7.

General reaction scheme 2

Formula (1) embodiment of the compounds according to the general reaction scheme (Method B) Preparation 2, wherein _{R a, R b, R c} , R d, R e, R f, R 1, R 2, R 3, A , E and Y are as defined above, and X represents chlorine, bromine, iodine or OTf. As shown in the general reaction scheme 2, intermediate A1 generates B1 in the presence of an oxidizing agent, B1 is hydrolyzed to B2 under alkaline conditions, B2 reacts with R ³ -YX fragments under appropriate conditions to generate B3, and B3 removes the protective group ( For example, Boc) obtains B4, B4 is coupled with R1-X to obtain B5, B5 is deprotected (for example, Cbz) to obtain B6, and B6 is reacted with acid chloride or acid anhydride compound to generate B7.

Further forms of the compound

"Pharmaceutically acceptable" here refers to a substance, such as a carrier or diluent, that does not make the biological activity or properties of the compound disappear, and is relatively non-toxic, for example, when a substance is administered to an individual, it does not cause unwanted biological effects or Interacts with any components it contains in a harmful way.

The term "pharmaceutically acceptable salt" refers to a form of a compound that does not cause important irritation to the administered organism and does not cause the biological activity and properties of the compound to disappear. In some specific aspects, pharmaceutically acceptable salts are obtained by reacting compounds of formula (1) with acids, such as hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, phosphoric acid, nitric acid, phosphoric acid and other inorganic acids, formic acid, acetic acid, Propionic acid, oxalic acid, trifluoroacetic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, picric acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, etc. Organic acids and acidic amino acids such as aspartic acid and glutamic acid.

It should be understood that references to pharmaceutically acceptable salts include solvent addition forms or crystalline forms, especially solvates or polymorphs. Solvates contain stoichiometric or non-stoichiometric solvents, and are selectively formed during crystallization with pharmaceutically acceptable solvents such as water, ethanol, etc. A hydrate is formed when the solvent is water, or an alcoholate is formed when the solvent is ethanol. Solvates of the compound of formula (1) are conveniently prepared or formed according to the methods described herein. For example, the hydrate of the compound of formula (1) is conveniently prepared by recrystallization from a mixed solvent of water/organic solvent. The organic solvent used includes, but is not limited to, dioxane, tetrahydrofuran, ethanol or methanol. In addition, the compounds mentioned here can exist in unsolvated and solvated forms. In summary, for the purposes of the compounds and methods provided herein, the solvated form is considered equivalent to the unsolvated form.

In other specific embodiments, the compound of formula (1) is prepared in different forms, including, but not limited to, amorphous, pulverized, and nano-particle size forms. In addition, the compound of formula (1) includes a crystalline form and may also be a polymorphic form. Polymorphs include different lattice arrangements of the same elemental composition of the compound. Polymorphs usually have different X-ray diffraction patterns, infrared spectra, melting points, density, hardness, crystal form, optical and electrical properties, stability and solubility. Different factors such as recrystallization solvent, crystallization rate and storage temperature may cause a single crystal form to dominate.

In another aspect, the compound of formula (1) has one or more stereocenters and is therefore in the form of racemates, racemic mixtures, single enantiomers, diastereomeric compounds and single diastereomers appear. The asymmetric centers that can exist depend on the nature of the various substituents on the molecule. Each such asymmetric center will independently produce two optical isomers, and all possible mixtures of optical isomers and diastereomers and pure or partially pure compounds are included within the scope of the present invention. The present invention is meant to include all such isomeric forms of these compounds.

the term

Unless otherwise stated, the terms used in the application of the present invention, including the specification and claims, are defined as follows. It must be noted that in the specification and appended claims, if the text does not clearly indicate otherwise, the singular form "a" includes the plural meaning. If not otherwise stated, conventional methods of mass spectrometry, nuclear magnetism, HPLC, protein chemistry, biochemistry, recombinant DNA technology and pharmacology are used. In this application, the use of "or" or "and" means "and/or" unless otherwise specified.

"Alkyl" refers to saturated aliphatic hydrocarbon groups, including straight and branched chain groups of 1 to 6 carbon atoms. Preferably, a lower alkyl group containing 1 to 4 carbon atoms, such as methyl, ethyl, propyl, 2-propyl, n-butyl, isobutyl, tert-butyl. As used herein, "alkyl" includes unsubstituted and substituted alkyl groups, especially alkyl groups substituted with one or more halogens. Preferred alkyl groups are selected from CH ₃ , CH ₃ CH ₂ , CF ₃ , CHF ₂ , CF ₃ CH ₂ , ⁱ Pr, ⁿ Pr, ⁱ Bu, ^c Pr, ⁿ Bu or ^t Bu.

"Cycloalkyl" refers to a 3- to 6-membered all-carbon monocyclic aliphatic hydrocarbon group in which one or more rings may contain one or more double bonds, but none of the rings have a fully conjugated π-electron system. For example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexane, cyclohexadiene and the like.

"Alkoxy" refers to an alkyl group bonded to the rest of the molecule through an ether oxygen atom. Representative alkoxy groups are those with 1-6 carbon atoms, such as methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, sec-butoxy And tert-butoxy. As used herein, "alkoxy" includes unsubstituted and substituted alkoxy, especially alkoxy substituted with one or more halogens. Preferred alkoxy groups are selected from OCH ₃ , OCF ₃ , CHF ₂ O, CF ₃ CH ₂ O, ⁱ PrO, ⁿ PrO, ⁱ BuO, ^c PrO, ⁿ BuO or ^t BuO.

"Aryl" refers to a group having at least one aromatic ring structure, that is, a carbocyclic aryl group having a conjugated π-electron system, such as a benzene ring and a naphthalene ring.

"Heteroaryl" refers to an aromatic group containing one or more heteroatoms (O, S or N). The heteroaryl group is monocyclic or polycyclic, such as a monocyclic heteroaryl ring and one or more carbocyclic rings. Aromatic groups or other monocyclic heterocyclic groups are condensed. Examples of heteroaryl groups include, but are not limited to, pyridyl, pyridazinyl, imidazolyl, pyrimidinyl, pyrazolyl, triazolyl, pyrazinyl, quinolinyl, isoquinolinyl, tetrazolyl, furanyl , Thienyl, isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, pyrrolyl, indolyl, benzimidazolyl, benzofuranyl, benzothiazolyl, benzothienyl, benzoxanyl Azolyl, benzopyridyl and pyrrolopyrimidinyl.

"Halogen" refers to fluorine, chlorine, bromine or iodine.

The term "bond" or "single bond" refers to a chemical bond between two atoms or between two fragments (when the atoms connected by a bond are considered part of a larger structure). On the one hand, when the group described herein is a bond, the lack of a reference group allows a bond to be formed between the remaining defined groups.

The term "membered ring" includes any cyclic structure. The term "element" means to indicate the number of skeletal atoms constituting the ring. Thus, for example, cyclohexyl, pyridyl, pyranyl, and thiopyranyl are six-membered rings, and cyclopentyl, pyrrolyl, furyl, and thienyl are five-membered rings.

The term "fragment" refers to a specific part or functional group of a molecule. Chemical fragments are generally considered to be chemical entities contained or attached to molecules.

Specific pharmacy and medical terms

The term "acceptable", as used herein, refers to a prescription component or active ingredient that does not have unduly harmful effects on the health of the general treatment target.

The terms "treatment", "treatment process" or "therapy" as used herein include alleviating, inhibiting, or improving the symptoms or conditions of diseases; inhibiting the occurrence of complications; improving or preventing underlying metabolic syndrome; inhibiting the occurrence of diseases or symptoms, Such as controlling the development of diseases or conditions; reducing diseases or symptoms; reducing diseases or symptoms; reducing complications caused by diseases or symptoms, or preventing or treating signs caused by diseases or symptoms.

As used herein, a certain compound or pharmaceutical composition, after administration, can improve a certain disease, symptom or condition, especially its severity, delay the onset, slow the progression of the disease, or reduce the duration of the disease. Regardless of fixed administration or temporary administration, continuous administration or intermittent administration, it can be attributed to or related to the administration.

"Active ingredient" refers to the compound represented by the general formula (1) and the pharmaceutically acceptable inorganic or organic salt of the compound of the general formula (1). The compounds of the present invention may contain one or more asymmetric centers, and therefore appear as racemates, racemic mixtures, single enantiomers, diastereomeric compounds, and single diastereomers. The asymmetric centers that can exist depend on the nature of the various substituents on the molecule. Each such asymmetric center will independently produce two optical isomers, and all possible mixtures of optical isomers and diastereomers and pure or partially pure compounds are included within the scope of the present invention. The present invention is meant to include all such isomeric forms of these compounds.

The terms "compound", "composition", "agent" or "medicine or medicament" are used interchangeably herein, and all refer to when administered to an individual (human or medicament). Animal), a compound or composition that can induce the desired pharmaceutical and/or physiological response through local and/or systemic effects.

The term "administered (administered, administrative or, administration)" herein refers to the direct administration of the compound or composition, or the administration of a prodrug, derivative, or analog of the active compound Etc., and can form an equivalent amount of the active compound in the subject of administration.

Although the numerical ranges and parameters used to define the wider range of the present invention are approximate numerical values, the relevant numerical values in the specific embodiments are presented here as accurately as possible. However, any value inherently inevitably contains the standard deviation due to individual test methods. Here, "about" usually means that the actual value is within plus or minus 10%, 5%, 1%, or 0.5% of a specific value or range. Alternatively, the term "about" means that the actual value falls within the acceptable standard error of the average, depending on the consideration of those skilled in the art. In addition to the experimental examples, or unless otherwise clearly stated, all ranges, quantities, values and percentages used herein (for example, used to describe the amount of material, length of time, temperature, operating conditions, quantity ratios and other similar Those) have been modified by "about". Therefore, unless otherwise stated to the contrary, the numerical parameters disclosed in this specification and the appended claims are approximate values and can be changed according to requirements. At least these numerical parameters should be understood as the indicated effective number of digits and the value obtained by the general carry method.

Unless otherwise defined in this specification, the scientific and technical terms used herein have the same meaning as the usual meaning understood by those skilled in the art. In addition, without conflict with context, the singular nouns used in this specification cover the plural nouns; and the plural nouns also cover the singular nouns.

Therapeutic use

The present invention provides methods of using the compounds or pharmaceutical compositions of the present invention to treat diseases, including but not limited to conditions involving G12C K-Ras, G12C H-Ras, and/or G12C N-Ras mutations (such as cancer).

In some factual solutions, a method for cancer treatment is provided, which method comprises administering to an individual in need an effective amount of any of the aforementioned protective structure (1) compound pharmaceutical compositions. In some embodiments, the cancer is mediated by K-Ras, H-Ras, and/or G12C N-Ras mutations. In other embodiments, the cancer is lung cancer, pancreatic cancer, colon cancer, MYH-related polyposis, or colorectal cancer.

Route of administration

The compound of the present invention and its pharmaceutically acceptable salt can be prepared into various preparations, which contain a safe and effective amount of the compound of the present invention or its pharmaceutically acceptable salt and a pharmacologically acceptable excipient or carrier . The "safe and effective amount" refers to: the amount of the compound is sufficient to significantly improve the condition without causing serious side effects. The safe and effective amount of the compound is determined according to the age, condition, and course of treatment of the subject to be treated.

"Pharmaceutically acceptable excipient or carrier" refers to: one or more compatible solid or liquid fillers or gel substances, which are suitable for human use, and must have sufficient purity and sufficiently low toxicity . "Compatibility" here means that each component of the composition can be blended with the compound of the present invention and between them without significantly reducing the efficacy of the compound. Examples of pharmacologically acceptable excipients or carriers include cellulose and its derivatives (such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants ( Such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerin, mannitol, sorbitol, etc.), emulsifiers (such as Tween)

), wetting agents (such as sodium lauryl sulfate), coloring agents, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

When the compound of the present invention is administered, it can be administered orally, rectally, parenterally (intravenous, intramuscular, or subcutaneous), or locally.

Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or mixed with the following ingredients: (a) fillers or compatibilizers, for example, Starch, lactose, sucrose, glucose, mannitol and silicic acid; (b) binders, such as hydroxymethyl cellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose and gum arabic; (c) humectant, For example, glycerin; (d) disintegrants, such as agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) slow solvents, such as paraffin; (f) Absorption accelerators, such as quaternary amine compounds; (g) wetting agents, such as cetyl alcohol and glyceryl monostearate; (h) adsorbents, such as kaolin; and (i) lubricants, such as talc, hard Calcium fatty acid, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, or mixtures thereof. In capsules, tablets and pills, the dosage form may also contain buffering agents.

Solid dosage forms such as tablets, sugar pills, capsules, pills and granules can be prepared with coatings and shell materials, such as enteric coatings and other materials known in the art. They may contain opacifying agents, and the active compound or the release of the compound in such a composition may be released in a certain part of the digestive tract in a delayed manner. Examples of embedding components that can be used are polymeric substances and waxes. If necessary, the active compound can also be formed into microcapsules with one or more of the above-mentioned excipients.

Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active compound, the liquid dosage form may contain inert diluents conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1 , 3-Butanediol, dimethylformamide and oils, especially cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil or mixtures of these substances.

In addition to these inert diluents, the composition may also contain adjuvants such as wetting agents, emulsifying and suspending agents, sweetening agents, flavoring agents and perfumes.

In addition to the active compound, the suspension may contain suspending agents, for example, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide and agar, or mixtures of these substances, and the like.

The composition for parenteral injection may contain physiologically acceptable sterile aqueous or non-aqueous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and non-aqueous carriers, diluents, solvents or excipients include water, ethanol, polyols and suitable mixtures thereof.

The dosage form of the compound of the present invention for topical administration includes ointment, powder, patch, spray and inhalant. The active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants that may be required if necessary.

The compound of the present invention can be administered alone or in combination with other pharmaceutically acceptable compounds.

When using the pharmaceutical composition, a safe and effective amount of the compound of the present invention is applied to a mammal (such as a human) in need of treatment, wherein the dosage is the pharmaceutically effective dosage considered to be administered. For a 60kg body weight, the daily The administration dose is usually 1 to 1000 mg, preferably 10 to 500 mg. Of course, the specific dosage should also consider factors such as the route of administration, the patient's health status, etc., which are within the skill range of a skilled physician.

The above-mentioned features mentioned in the present invention or the features mentioned in the embodiments can be combined arbitrarily. All the features disclosed in the specification of this case can be used in combination with any composition form, and each feature disclosed in the specification can be replaced by any alternative feature that can provide the same, equal or similar purpose. Therefore, unless otherwise specified, the disclosed features are only general examples of equal or similar features.

In the following description, various specific aspects, characteristics and advantages of the above-mentioned compounds, methods, and pharmaceutical compositions will be described in detail to make the content of the present invention very clear. It should be understood that the following detailed description and examples describe specific embodiments and are for reference only. After reading the description of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent situations also fall within the scope defined by this application.

In all the examples, ¹ H-NMR was recorded with a Vian Mercury 400 nuclear magnetic resonance instrument, and the chemical shifts were expressed in δ (ppm); the silica gel used for separation was not specified, all were 200-300 meshes, and the ratio of the eluate was volume ratio.

The present invention uses the following acronyms: Ar stands for argon; aq stands for aqueous solution; (Boc) ₂ O stands for di-tert-butyl dicarbonate; CDCl ₃ stands for deuterated chloroform; CD ₃ OD stands for deuterated methanol; CH ₃ CN stands for Acetonitrile; CH ₃ NO ₂ stands for nitromethane; (COCl) ₂ stands for oxalyl chloride; Cs ₂ CO ₃ stands for cesium carbonate; CuI stands for cuprous iodide; DBU stands for 1,8-diaza heterocycle [5,4,0 ]Undecene-7; DCM stands for dichloromethane; DIPEA stands for diisopropylethylamine; Dioxane stands for 1,4-dioxane; DMF stands for dimethylformamide; DMSO stands for dimethyl sulfoxide; EA stands for ethyl acetate; EtOH stands for ethanol; EtONa stands for sodium ethoxide; h stands for hours; NaOH stands for sodium hydroxide; NaOMe stands for sodium methoxide; LC-MS stands for liquid phase-mass spectrometry; m-CPBA stands for m-chloroperoxybenzoic acid; MeOH stands for methanol; min stands for minutes; MS stands for mass spectrometry; Na ₂ CO ₃ stands for sodium carbonate; NaH stands for sodium hydride; NMR stands for nuclear magnetic resonance; Pd ₂ (dba) ₃ stands for tris(dibenzylideneacetone) dipalladium; PE stands for Petroleum ether; SOCl ₂ stands for thionyl chloride; ^t- BuOK stands for potassium tert-butoxide; ^t- BuONa stands for sodium tert-butoxide; TFA (CF ₃ COOH) stands for trifluoroacetic acid; Tf ₂ O stands for trifluoromethanesulfonic anhydride; THF stands for tetrahydrofuran; Xantphos stands for 4,5-bis(diphenylphosphine)-9,9-dimethylxanthene.

detailed description

Preparation Example 1: 2'-(methylthio)-4'-(((trifluoromethyl)sulfonyl)oxy)-5',8'-dihydro-7'H-spiro[cyclopropane-1 Synthesis of ,6'-pyrido[3,4-d]pyrimidine]-7'-carboxylic acid tert-butyl ester (S-1)

(E)-3-(1-((tert-butoxycarbonyl)amino)cyclopropyl)methyl acrylate

Add methyl 2-(diethoxyphosphoryl)acetate (21.0g, 0.1mol) to a 500mL three-necked flask, dry THF (200mL), and reduce the temperature to -20℃ under Ar gas protection, and then at -20℃～-10 LiHMDS (130mL, 1M in THF, 0.13mol) was added dropwise at ℃, after dripping, the mixture was stirred at -10℃ for 30min, and then (1-aldehyde cyclopropyl) t-butyl carbamate (16.7) was added dropwise at -10℃ g, 90mmol) in THF (30mL), after dripping, the mixture is stirred at -10°C~rt for 5h. After the completion of the reaction monitored by LC-MS, it was quenched by adding saturated ammonium chloride solution (100mL), extracted with EA (100mL), the organic phase was washed with saturated sodium chloride solution (100mL), concentrated to dryness, and the residue was column chromatographed (EA /PE=0/5 to 1/5) was purified to obtain a light brown oil (13.46g, yield: 62%), ESI-MS m/z: 242.0 [M+H] ⁺ .

(E)-3-(1-((tert-butoxycarbonyl)(2-methoxy-2-oxoethyl)amino)cyclopropyl)methyl acrylate

Add (E)-3-(1-((tert-butoxycarbonyl)amino)cyclopropyl)methyl acrylate (13.4g, 55.5mmol), DMF (135mL), Ar gas protection and reduce the temperature to After 0℃, NaH (2.89g, 60% in mineral, 72.2mmol) was added in batches. After the addition, the system was stirred at 0～5℃ for 30min. Then ethyl bromoacetate (10.2g, 66.6mmol) was added. The mixture was stirred at room temperature for reaction. 20h. After the reaction was monitored by LC-MS, the system was quenched with saturated ammonium chloride solution (100mL), extracted with EA (100mL*2), the combined organic phases were washed with saturated sodium chloride solution (100mL), concentrated to dryness, and the residue column Purification by chromatography (EA/PE=0/5 to 1/5) gave a light brown oil (8.87g, yield 51%), ESI-MS m/z: 336.0 [M+Na] ⁺ .

Methyl 3-(1-((tert-butoxycarbonyl)(2-methoxy-2-oxoethyl)amino)cyclopropyl)propionate

Add (E)-3-(1-((tert-butoxycarbonyl)(2-methoxy-2-oxoethyl)amino)cyclopropyl)methyl acrylate (8.87g, 28.34) to a 500mL single-mouth bottle. mmol), MeOH (170 mL), 10% Pd/C (800 mg, wet% = 50%), H _{2 was} replaced three times and then hydrogenated under normal pressure at room temperature for 20 h. After the completion of the reaction monitored by LC-MS, the system was filtered through celite, and the filtrate was concentrated to dryness to obtain a colorless oil (9.1 g, yield 100%), ESI-MS m/z: 338.0 [M+Na] ⁺ .

4-(tert-butyl)7-ethyl 6-oxo-4-azaspiro[2.5]octane-4,7-dicarboxylate

Methyl 3-(1-((tert-butoxycarbonyl)(2-methoxy-2-oxoethyl)amino)cyclopropyl)propionate (9.1g, 28.34mmol) was dissolved in anhydrous methanol (180mL), add freshly prepared sodium methoxide (2.30g, 42.51mmol) under the protection of Ar, then heat to reflux for 5h, LC-MS monitors the completion of the reaction, adjust the pH of the mixture to 5-6, concentrate, and add acetic acid to the residue Ethyl acetate (100 mL), H ₂ O (100 mL), the aqueous phase was extracted with EA (50 mL), the combined organic phases were washed with saturated sodium chloride solution (50 mL), concentrated, and column chromatography (PE/EA=10/1) to 2/1) Purification to obtain a nearly colorless oil (3.3g, yield 41%), ESI-MS m/z: 284.0[M+H] ⁺ .

2'-(Methylthio)-4'-oxo-3',4',5',8'-tetrahydro-7'H-spiro[cyclopropane-1,6'-pyrido[3,4 -d]]pyrimidine]-7'-tert-butyl carboxylate

Add 4-(tert-butyl)7-ethyl 6-oxo-4-azaspiro[2.5]octane-4,7-dicarboxylate (3.3g, 11.65mmol) and MeOH into a 250mL single-mouth bottle (66 mL), sodium methoxide (3.14 g, 58.2 mmol) and S-methyl isothiourea sulfate (3.5 g, 17.5 mmol) were added under the protection of Ar, and reacted at room temperature for 20 h. Monitoring by LC-MS and TLC (PE/EA=2/1), after the reaction was completed, the mixture was adjusted to pH 5-6 with 2N HCl in an ice bath, and concentrated under reduced pressure. H ₂ O (60 mL) and EA (15 mL) were added to the residue, the slurry was stirred for half an hour, filtered, and dried to obtain an off-white solid (3.43 g, yield 91%), ESI-MS m/z: 324.1 [M+ H] ⁺ .

2'-(methylthio)-4'-(((trifluoromethyl)sulfonyl)oxy)-5',8'-dihydro-7'H-spiro[cyclopropane-1,6'- Pyrido[3,4-d]pyrimidine]-7'-carboxylic acid tert-butyl ester

Add 2'-(methylthio)-4'-oxo-3',4',5',8'-tetrahydro-7'H-spiro[cyclopropane-1,6'- into a 250mL three-necked flask Pyrido[3,4-d]]pyrimidine]-7'-carboxylic acid tert-butyl ester (3.43g, 10.6mmol), DIPEA (4.1g, 31.8mmol) and DCM (70mL), protected by Ar, cooled to At 0-5°C, a solution of Tf ₂ O (3.89 g, 13.78 mmol) in DCM (20 mL) was added dropwise. After the addition was completed, the mixture was stirred at room temperature and reacted for 20 hours. TLC monitoring (PE/EA=2/1) After the reaction is complete, add H ₂ O (100 mL) to quench, then add 1N HCl dropwise to adjust the pH to 5-6, separate the liquids, and use saturated sodium chloride solution (50 mL ) Wash, concentrate, and purify the residue by column chromatography (PE/EA=10/0 to 5/1) to obtain an off-white solid (3.5g, yield 72.6%), ESI-MS m/z: 456.1[M+H ] ⁺ .

Preparation Example 2: 2'-(methylthio)-4'-(((trifluoromethyl)sulfonyl)oxy)-5',8'-dihydro-7'H-spiro[cyclobutane- Synthesis of 1,6'-pyrido[3,4-d]pyrimidine]-7'-carboxylic acid tert-butyl ester (S-2)

Using tert-butyl (1-aldehyde cyclobutyl) carbamate as raw material, the target intermediate S-2 was obtained by the synthesis method of intermediate S-1, ESI-MS m/z: 470.1 [M+H] ⁺ .

Preparation Example 3: 2'-(methylthio)-4'-(((trifluoromethyl)sulfonyl)oxy)-5',8'-dihydro-7'H-spiro[oxetan Synthesis of alkane-3,6'-pyrido[3,4-d]pyrimidine]-7'-carboxylic acid tert-butyl ester (S-3)

Using tert-butyl (3-formyloxetan-3-yl)carbamate as raw material, the target intermediate S-3 was obtained by the synthesis method of intermediate S-1, ESI-MS m/z:472.1[ M+H] ⁺ .

Preparation Example 4: 2'-(methylthio)-4'-(((trifluoromethyl)sulfonyl)oxy)-5',6'-dihydro-7'H-spiro[cyclopropane-1 Synthesis of ,8'-pyrido[3,4-d]pyrimidine]-7'-carboxylic acid tert-butyl ester (S-4)

Ethyl 1-aminocyclopropane-1-carboxylate

Dissolve 1-aminocyclobutane-1-carboxylic acid (10.1g, 0.1mol) in ethanol (100mL), slowly add thionyl chloride (23.8g, 0.2mol) dropwise under ice bath, and react at room temperature for 20h after the completion , LC-MS monitored the completion of the reaction, the mixture was concentrated, the residue was added with DCM (100mL) and sodium bicarbonate aqueous solution (100mL), stirred, separated, the aqueous phase was extracted with (100mL*2), and the organic phase was combined with saturated chlorine Wash with sodium chloride solution (100 mL*2), dry with anhydrous sodium sulfate, filter, and concentrate to obtain a pale yellow oil (9.08 g, yield 70%), ESI-MS m/z: 130.1 [M+H] ⁺ .

Ethyl 1-((4-ethoxy-4-oxobutyl)amino)cyclopropane-1-carboxylate

Dissolve ethyl 1-aminocyclopropane-1-carboxylate (9.08g, 70.4mmol) in acetonitrile (200mL), add DIPEA (18.2g, 141mmol), ethyl 4-bromobutyrate (27.5g, 141mmol) , Under the protection of Ar, the temperature was raised to 80°C and reacted for 20h. LC-MS monitored the completion of the reaction. After concentration, the residue was added with ethyl acetate (200mL), H ₂ O (100mL), stirred and separated. The aqueous phase was separated with ethyl acetate ( 100mL), the combined organic phase was washed with saturated sodium chloride (100mL), concentrated, and the residue was purified by column chromatography (DCM/MeOH=20/0 to 20/1) to obtain a light yellow oil (8.70g, yield 51 %), ESI-MS m/z: 244.2 [M+H] ⁺ .

Ethyl 1-((tert-butoxycarbonyl)(4-ethoxy-4-oxobutyl)amino)cyclopropane-1-carboxylate

Dissolve 1-((4-ethoxy-4-oxobutyl)amino)cyclopropane-1-carboxylic acid ethyl ester (8.70g, 35.8mmol) with THF (100mL) and H ₂ O (50mL), Boc ₂ O (15.7g, 72mmol) and Na ₂ CO ₃ (11.4g, 0.107mol) were added, the mixture was raised to 60℃ and the reaction was stirred for 20 hours. After the completion of the reaction was monitored by LC-MS, the reaction solution was added with EA (100mL) and H ₂ O (100mL), stirring, liquid separation, the organic phase was concentrated, the residue was purified by column chromatography (PE/EA=5/0 to 5/1) to obtain a colorless oil (7.61g, yield 62%), ESI-MS m/z: 344.2 [M+H] ⁺ .

4-(tert-butyl)7-ethyl-8-oxo-4-azaspiro[2.5]octane-4,7-dicarboxylate

Dissolve 1-((tert-butoxycarbonyl)(4-ethoxy-4-oxobutyl)amino)cyclopropane-1-carboxylic acid ethyl ester (7.61g, 22.2mmol) in absolute ethanol (140mL) Under the protection of Ar, freshly prepared sodium ethoxide (2.26g, 33.3mmol) was added, and then the temperature was raised to reflux for 5h. LC-MS monitored the completion of the reaction. The pH of the mixture was adjusted to 5-6, concentrated, and the residue was added with ethyl acetate ( 100mL) and H ₂ O (100mL), the aqueous phase was extracted with EA (50mL), the combined organic phases were washed with saturated sodium chloride solution (50mL), concentrated, and column chromatography (PE/EA=10/1 to 2/ 1) Purification to obtain a nearly colorless oil (2.31g, yield 35%), ESI-MS m/z: 298.0[M+H] ⁺ .

2'-(Methylthio)-4'-carbonyl-3',4',5',6'-tetrahydro-7'H-spiro[cyclopropane-1,8'-pyrido[3,4- d]pyrimidine]-7'-tert-butyl carboxylate

Add 4-(tert-butyl)7-ethyl-8-oxo-4-azaspiro[2.5]octane-4,7-dicarboxylate (4.76g, 16.0mmol) and Sodium ethoxide (5.44g, 80mmol) and S-methylisothiourea sulfate (4.8g, 24mmol) were added under the protection of EtOH (100mL) and Ar, respectively, and reacted at room temperature for 20h. Monitoring by LC-MS and TLC (PE/EA=2/1), after the reaction was completed, the mixture was adjusted to pH 5-6 with 2N HCl in an ice bath, and concentrated under reduced pressure. H ₂ O (60 mL) and EA (30 mL) were added to the residue, the slurry was stirred for half an hour, filtered, and dried to obtain an off-white solid (4.24 g, yield 82%), ESI-MS m/z: 324.1 [M+ H] ⁺ .

2'-(methylthio)-4'-(((trifluoromethyl)sulfonyl)oxy)-5',6'-dihydro-7'H-spiro[cyclopropane-1,8'- Pyrido[3,4-d]pyrimidine]-7'-carboxylic acid tert-butyl ester

Add 2'-(methylthio)-4'-carbonyl-3',4',5',6'-tetrahydro-7'H-spiro[cyclobutane-1,8'- into a 250mL three-necked flask Pyrido[3,4-d]pyrimidine]-7'-carboxylic acid tert-butyl ester (4.24g, 13.11mmol), DIPEA (5.08g, 39.40mmol) and DCM (100mL), protected by Ar, cooled to At 0-5°C, a solution of Tf ₂ O (4.45 g, 15.75 mmol) in DCM (20 mL) was added dropwise. After the addition was completed, the mixture was stirred and reacted at room temperature for 20 hours. TLC monitoring (PE/EA=2/1) After the reaction is complete, add H ₂ O (100 mL) to quench, then add 1N HCl dropwise to adjust the pH to 5-6, separate the liquids, and use saturated sodium chloride solution (50 mL ) Wash, concentrate, and purify the residue by column chromatography (PE/EA=10/0 to 10/1) to obtain an off-white solid (4.35g, yield 73%), ESI-MS m/z: 456.1[M+H ] ⁺ .

Preparation Example 5: 2'-(methylthio)-4'-(((trifluoromethyl)sulfonyl)oxy)-5',6'-dihydro-7'H-spiro[cyclobutane- Synthesis of 1,8'-pyrido[3,4-d]pyrimidine]-7'-carboxylic acid tert-butyl ester (S-5)

Using 1-aminocyclobutyl-1-carboxylic acid as a raw material, the target intermediate S-5 was obtained by the synthetic method of intermediate S-4, ESI-MS m/z: 470.1 [M+H] ⁺ .

Preparation Example 6: 2'-(methylthio)-4'-(((trifluoromethyl)sulfonyl)oxy)-5',6'-dihydro-7'H-spiro[oxetan Synthesis of alkane-1,8'-pyrido[3,4-d]pyrimidine]-7'-carboxylic acid tert-butyl ester (S-6)

Using 3-aminooxetane-3-carboxylic acid as a raw material, the target intermediate S-6 was obtained by the synthesis method of fragment S-4, ESI-MS m/z: 472.1 [M+H] ⁺ .

Preparation Example 7: 2'-(methylthio)-4'-(((trifluoromethyl)sulfonyl)oxy)-6'H-spiro[oxetane-3,5'-pyrido Synthesis of [3,4-d]pyrimidine]-7'(8'H)-tert-butyl carboxylate (S-7)

2-(oxetan-3-ylidene) ethyl acetate

Dissolve oxetane-3-one (10.8g, 150mm0l) in dichloromethane (150mL), add ethoxyformylmethylenetriphenylphosphonium (52.2g, 150mmol) under ice bath, add After completion, react at room temperature overnight. LC-MS monitors the completion of the reaction. Concentrate to obtain a white solid. The solid is slurried with PE/EA (10/1, 100mL), the filtrate is concentrated, and column chromatography (PE/EA=20/1 to 5/1) , A nearly colorless gum (15g, yield 70%) was obtained, ESI-MS m/z: 143.0[M+H] ⁺ .

2-(3-(nitromethyl)oxetan-3-yl)ethyl acetate

Dissolve 2-(oxetan-3-ylidene) ethyl acetate (14.2g, 100mL) in nitromethane (100mL), add DBU (1.52g, 10mmol), react at room temperature overnight, LC-MS Monitor the completion of the reaction, concentrate, dissolve the residue with ethyl acetate, wash with dilute acid, concentrate and column chromatography (PE/EA=20/1to 5/1) to obtain a nearly colorless gum (10g, yield 50%) ), ESI-MS m/z: 204.0 [M+H] ⁺ .

2-(3-(aminomethyl)oxetan-3-yl)ethyl acetate

Dissolve 2-(3-(nitromethyl)oxetan-3-yl)ethyl acetate (6.1g, 30mmol) in ethanol (100mL), add trifluoroacetic acid (6.8g, 60mmol), Add 10% Pd/C (1.36g, wet%=20%), add H ₂ , react overnight at 50°C, monitor by LC-Ms, after the reaction is complete, filter, and concentrate the initial product directly into the next reaction, ESI- MS m/z: 174.1 [M+H] ⁺ .

N-(tert-Butoxycarbonyl)-N-((3-(2-ethoxy-2-carbonylethyl)oxetan-3-yl)methyl)glycine ethyl ester

2-(3-(Aminomethyl)oxetan-3-yl)ethyl acetate (theoretical amount, 30mmol) was dissolved in acetonitrile (150mL), ethyl bromoacetate (5.0g, 30mmol) was added, Potassium carbonate (20g, 150mmol), heated at 50°C and reacted overnight. LC-MS monitoring, the reaction was completed, filtered, concentrated and dissolved in dichloromethane (150mL), DIPEA (7.74g, 60mmol), Boc ₂ O (7.8g) , 36mmol), react at room temperature overnight, monitored by LC-MS, the reaction is complete, the reaction solution is washed with water, concentrated and column chromatography (PE/EA = 20/1 to 5/1) to obtain a nearly colorless gum (5.4g, Yield 50%), ESI-MS m/z: 360.1 [M+H] ⁺ .

6-(tert-butyl)9-ethyl 8-carbonyl-2-oxa-6-azaspiro[3.5]nonane-6,9-dicarboxylate

Add N-(tert-butoxycarbonyl)-N-((3-(2-ethoxy-2-carbonylethyl)oxetan-3-yl)methyl)glycine ethyl ester (7.2g , 20mmol) was dissolved in absolute ethanol (60mL), and freshly prepared sodium ethoxide (2.7g, 40mmol) was added. The temperature was heated and refluxed for 5h, monitored by LC-MS. After the reaction, the pH was adjusted to neutral, concentrated, and ethyl acetate was added. (100mL) dissolved, washed with water, saturated sodium chloride solution, concentrated and column chromatography (PE/EA=10/1 to 2/1) to obtain a nearly colorless gum (2.5g, yield 40%), ESI-MS m/z: 314.1 [M+H] ⁺ .

2'-(Methylthio)-4'-carbonyl-4',8'-dihydro-3'H-spiro[oxetane-3,5'-pyrido[3,4-d]pyrimidine ]-7'(6'H)-tert-butyl formate

Add 6-(tert-butyl)9-ethyl 8-carbonyl-2-oxa-6-azaspiro[3.5]nonane-6,9-dicarboxylate (5g, 15.96mmol ) And EtOH (100 mL), under the protection of Ar, sodium ethoxide (5.43 g, 79.8 mmol) and S-methyl isothiourea sulfate (4.8 g, 23.9 mmol) were added respectively, and reacted at room temperature overnight. LC-MS monitoring and TLC spot plate (PE/EA=2/1), cooling in an ice bath, adjust the pH to 5 with 2N HCl, concentrate, add H ₂ O (30 mL) and EA (30 mL) to the crude product, beat and stir After half an hour, filtered and dried to obtain an off-white solid (4.3g, yield 79%), ESI-MS m/z: 340.1[M+H] ⁺ .

2'-(methylthio)-4'-(((trifluoromethyl)sulfonyl)oxy)-6'H-spiro[oxetane-3,5'-pyrido[3,4 -d]pyrimidine]-7'(8'H)-tert-butyl carboxylate

Add 2'-(methylthio)-4'-carbonyl-4',8'-dihydro-3'H-spiro[oxetane-3,5'-pyrido[3 ,4-d]pyrimidine]-7'(6'H)-carboxylic acid tert-butyl ester (4.0g, 11.80mmol), DIPEA (4.57g, 35.40mmol) and DCM (50mL), protected by Ar, cooled in an ice bath To 0-5°C, add Tf ₂ O (5.0 g, 17.7 mmol) in DCM (10 mL) solution dropwise, after the dropwise addition is complete, react at room temperature, TLC plate (PE/EA = 2/1), after the raw materials are basically reacted After concentrating the reaction solution, it was purified by column chromatography (PE/EA=1/0 to 10/1) to obtain a yellow solid (3.3g, yield 60%), ESI-MS m/z: 472.0[M+H] ⁺ .

Preparation Example 8: 2'-(methylthio)-4'-(((trifluoromethyl)sulfonyl)oxy)-6'H-spiro[cyclobutane-1,5'-pyrido[3 Synthesis of ,4-d]pyrimidine]-7'(8'H)-tert-butyl carboxylate (S-8)

Using cyclobutanone as raw material, the target intermediate S-8 was obtained by the synthesis method of fragment S-7, ESI-MS m/z: 470.1 [M+H] ⁺ .

Preparation Example 9: 2'-(methylthio)-4'-(((trifluoromethyl)sulfonyl)oxy)-6'H-spiro[cyclopropane-1,5'-pyrido[3, Synthesis of tert-butyl 4-d]pyrimidine]-7'(8'H)-carboxylate (S-9)

Using 2-cyclopropylmethylene ethyl acetate as raw material, the target intermediate S-9 was obtained by the synthesis method of fragment S-7, ESI-MS m/z: 456.1 [M+H] ⁺ .

Example 1: 2-((S)-1-acryloyl-4-(2'-(((S)-1-methylpyrrolidin-2-yl)methoxy)-7'-(naphthalene- 1-yl)-7',8'-dihydro-5'H-spiro[cyclopropane-1,6'-pyrido[3,4-d]pyrimidine]-4'-yl)piperazine-2- Synthesis of acetonitrile (compound 1)

Compound 1 was prepared according to Method A as described below:

(S)-4'-(4-((Benzyloxy)carbonyl)-3-(cyanomethyl)piperazin-1-yl)-2'-(methylthio)-5',8'- Dihydro-7'H-spiro[cyclopropane-1,6'-pyrido[3,4-d]pyrimidine]-7'-carboxylic acid tert-butyl ester (1-1)

Add S-1 (9.1g, 20.0mmol), DIPEA (5.16g, 40mmol), benzyl(S)-2-(cyanomethyl)piperazine-1-carboxylic acid benzyl ester (5.22 g, 20.0mmol) and DMF (91mL), under the protection of Ar, the reaction was heated to 100°C for 1 hour, monitored by TLC (PE/EA=10/1), the reaction of the raw materials was completed, the reaction solution was cooled to room temperature, and water ( 100mL), extracted with EA (100mL*2), combined the organic phases, washed with saturated sodium chloride solution, concentrated the organic phase, purified by column chromatography (PE/EA=1/0 to 2/1) to obtain a white solid 1-1 (10.6 g, yield 94%), ESI-MS m/z: 565.3 [M+H] ⁺ .

(S)-4'-(4-((Benzyloxy)carbonyl)-3-(cyanomethyl)piperazin-1-yl)-2'-(methylsulfonyl)-5',8' -Dihydro-7'H-spiro[cyclopropane-1,6'-pyrido[3,4-d]pyrimidine]-7'-carboxylic acid tert-butyl ester (1-2)

Add 1-1 (10.6g, 18.8mmol) and DCM (210mL) to a 500mL single-necked flask, under Ar protection, cool to 0-5℃ in an ice bath, add m-CPBA (11.4g, 65.8mmol), under ice bath React for 2 hours. TLC monitoring (PE/EA=1/5), the reaction of the raw materials is complete, add saturated sodium bicarbonate solution (60mL) to the reaction solution, stir and separate, the aqueous phase is extracted with DCM (100mL*2), and the organic phases are combined , Washed with saturated sodium chloride solution, concentrated the organic phase, and purified by column chromatography (PE: EA=1:0 to 1:1) to obtain white solid 1-2 (9.1g, yield 81%), ESI-MS m/ z:597.2[M+H] ⁺ .

4'-((S)-4-((benzyloxy)carbonyl)-3-(cyanomethyl)piperazin-1-yl)-2'-(((S)-1-methylpyrrolidine -2-yl)methoxy-tert-butyl)-5',8'-dihydro7'H-spiro[cyclopropane-1,6'-pyrido[3,4-d]pyrimidine]-7'- Tert-Butyl formate (1-3)

Add (S)-(1-methylpyrrolidin-2-yl)methanol (2.108g, 18.30mmol) and toluene (180mL) into a 500mL single-neck bottle, and cool to 0～5℃ in an ice bath under the protection of Ar gas, then Potassium tert-butoxide (2.57g, 22.88mmol) was added, after stirring at 0～5℃ for 15min, 1-2 (9.1g, 15.25mmol) was added, and the mixture was stirred at 0～10℃ for 2h. Monitoring by LC-MS and TLC (PE/EA=1/1), after the reaction of the raw materials is complete, the system is quenched by adding saturated ammonium chloride (100mL), stirred and separated, and the aqueous phase is extracted with EA (90mL*2). The combined organic phase was washed with saturated sodium chloride solution (90mL*2), concentrated, and the residue was purified by column chromatography (EA/PE=0/1 to 1/2) to obtain off-white solid 1-3 (7.23g, yield Rate 75%), ESI-MS m/z: 632.3 [M+H] ⁺ .

(S)-2-(cyanomethyl)-4-(2'-(((S)-1-methylpyrrolidin-2-yl)methoxy)-7',8'-dihydro- 5'H-spiro[cyclopropane-1,6'-pyrido[3,4-d]pyrimidine]-4'-yl)piperazine-1-carboxylic acid benzyl ester (1-4)

1-3 (7.23g, 11.44mmol), DCM (140mL) and TFA (25.2g, 221mmol) were added to a 250mL single-neck flask, and the mixture was stirred at room temperature for reaction for 2h. After the LC-MS monitoring the reaction was complete, the system was concentrated, the residue was added with DCM (200mL), sodium bicarbonate solution (100mL), stirred at room temperature for 30min and then separated, the aqueous phase was extracted with DCM (100mL), and the organic phase was combined with saturated chlorine Washed with sodium chloride solution, dried with anhydrous sodium sulfate, filtered and concentrated to dryness to obtain yellow-brown oil 1-4 (5.66g, yield 93%), ESI-MS m/z: 532.3[M+H] ⁺ .

(S)-2-(cyanomethyl)-4-(2'-(((S)-1-methylpyrrolidin-2-yl)methoxy)-7'-(naphthalene-1-yl )-7',8'Dihydro-5'H-spiro[cyclopropane-1,6'-pyrido[3,4-d]pyrimidine]-4'-yl)piperazine-1-carboxylic acid benzyl ester ( 1-5)

Add 1-4 (266mg, 0.5mmol), 1-bromonaphthalene (166mg, 0.75mmol), ^t- BuONa (144mg, 1.5mmol) and Dioxane (20mL) to a 100mL single-mouth bottle, add Sphos-G3 after Ar replacement protection -Pd (87mg, 0.1mmol), the mixture was heated to reflux and stirred for 20h under the protection of Ar. After the LC-MS monitoring reaction was completed, the system was quenched with water, extracted with EA (20mL*2), the organic phase was washed with saturated sodium chloride, concentrated and purified by column chromatography (DCM/MeOH/NH ₄ OH=40/1/0.02 ) To obtain light brown oil 1-5 (72 mg, yield 22%), ESI-MS m/z: 658.4 [M+H] ⁺ .

2-((S)-4-(2'-(((S)-1-methylpyrrolidin-2-yl)methoxy)-7'-(naphthalen-1-yl)-7',8 '-Dihydro-5'H-spiro[cyclopropane-1,6'-pyrido[3,4-d]pyrimidine]-4'-yl)piperazin-2-yl)acetonitrile (1-6)

Add 1-5 (72mg, 0.11mmol), MeOH (10mL) and 10% Pd/C (20mg, wet%=50%) to a 50mL single-mouth flask, and stir for 20h at room temperature and normal pressure after H ₂ replacement for three times. After the LC-MS monitoring reaction was completed, the system was filtered, and the filtrate was concentrated to obtain a yellow foamy solid 1-6 (58 mg, yield 100%), ESI-MS m/z: 524.3 [M+H] ⁺ .

2-((S)-1-acryloyl-4-(2'-(((S)-1-methylpyrrolidin-2-yl)methoxy)-7'-(naphthalene-1-yl) -7',8'-Dihydro-5'H-spiro[cyclopropane-1,6'-pyrido[3,4-d]pyrimidine]-4'-yl)piperazin-2-yl)acetonitrile ( Compound 1)

Add 1-6 (58mg, 0.11mmol), DCM (5mL), DIPEA (42mg, 0.33mmol) to a 50mL single-mouth flask, and cool to 0℃ in an ice bath under the protection of Ar, and then add acryloyl chloride (9mg, 0.10mmol) dropwise. After dripping the DCM solution, the system was stirred at 0-5°C for 1h. After the LC-MS monitors the completion of the reaction, it is quenched by adding water and separated. The aqueous phase is extracted with DCM (10 mL). The combined organic phases are washed with saturated sodium chloride solution (10 mL). After concentration, the residue is purified by pre-TLC to obtain an off-white color. Solid compound 1 (13 mg, yield 20%).

¹ H NMR (400MHz, CDCl ₃ ) δ: 7.97 (m, 1H), 7.72-7.21 (m, 6H), 6.63-6.52 (m, 1H), 6.24 (dd, J = 16.9, 1.9 Hz, 1H), 5.72 (ddd, J = 10.5, 3.2, 1.9 Hz, 1H), 4.66 (d, J = 11.9 Hz, 1H), 4.32 (m, 3H), 3.64-3.51 (m, 3H), 3.48-3.22 (m, 4H),2.74(s,3H),2.53-2.32(m,7H),1.86-1.65(m,4H),0.65-0.46(m,4H); ESI-MS m/z:578.3[M+H] ⁺ .

Example 2-Example 93: Synthesis of Compound 2-Compound 93

Using intermediates S-1 to S-9 as starting materials, using synthetic method A, similar to the synthetic method of compound 1, compound 2-compound 93 can be obtained.

Example 94: 2-((S)-1-(2-fluoroacryloyl)-4-(7'-(8-methylnaphthalene-1-yl)-1'-(((S)-1- (Methylpyrrolidin-2-yl)methyl)-2'-oxy-1',5',7',8'-tetrahydro-2'H-spiro[cyclopropane-1,6'-pyrido[ Synthesis of 3,4-d]pyrimidine)-4'-yl)piperazin-2-yl)acetonitrile (compound 94)

Compound 94 was prepared according to Method B as described below:

(S)-4'-(4-((Benzyloxy)carbonyl)-3-(cyanomethyl)piperazin-1-yl)-2'-oxo-1',2',5', 8'-Tetrahydro-7'H-spiro[cyclopropane-1,6'-pyrido[3,4-d]pyrimidine]-7'-carboxylic acid tert-butyl ester (94-1)

Add 1-2 (5.27g, 8.83mmol) and 1,4-dioxane (100mL) to a 250mL single-mouth flask, then add sodium hydroxide solution (44.1mL, 2N), react at room temperature overnight, TLC monitoring (PE /EA = 1/1) After the raw material has reacted completely, adjust the pH to 7 with 2N hydrochloric acid to separate out a solid, filter to obtain a white solid 94-1, and then extract the filtrate with EA (50mL), wash with saturated sodium chloride solution, and anhydrous sulfuric acid The sodium was dried and filtered. After concentration, the crude product was added to EA (5mL) to be slurried and stirred, filtered to obtain a white solid 94-1, which was combined to obtain the product 94-1 (3.82g, yield 81%), ESI-MS m/z: 535.2 [M +H] ⁺ .

4'-((S)-4-((benzyloxy)carbonyl)-3-(cyanomethyl)piperazin-1-yl)-1'-(((S)-1-methylpyrrolidine -2-yl)methyl)-2'-oxo-1',2',5',8'-tetrahydro-7'-spiro[cyclopropane-1,6'-pyrido[3,4- d]pyrimidine)-7'-tert-butyl carboxylate (94-2)

Add 94-1 (1.26g, 2.36mmol), Cs ₂ CO ₃ (1.53g, 4.70mmol), (S)-2-(bromomethyl)-1-methylpyrrolidine (632mg) to the 100mL sealed tube reactor , 3.55mmol), CuI (89mg, 0.47mmol) and DMSO (24mL), after sealing, the temperature was raised to 100°C and the reaction was stirred for 20h. The system was cooled to room temperature, quenched by adding saturated ammonium chloride solution (20mL), stirred at room temperature for 30min and then extracted with EA (20mL*2), the combined organic phase was washed with saturated sodium chloride solution (20mL*2), concentrated, and the residue Purified by column chromatography (DCM/MeOH=30/0 to 20/1) to obtain a yellow-brown solid 94-2 (610 mg, yield 41%), ESI-MS m/z: 632.3 [M+H] ⁺ .

(S)-2-(cyanomethyl)-4-(1'-(((S)-1-methylpyrrolidin-2-yl)methyl)-2'-oxo-1',5 ',7',8'-Tetrahydro-2'H-spiro[cyclopropane-1,6'-pyrido[3,4-d]pyrimidine]-4'-yl)piperazine-1-carboxylic acid benzyl ester (94-3)

94-2 (610 mg, 0.966 mmol), DCM (10 mL) and TFA (2.20 g, 19.31 mmol) were added to a 250 mL single-neck flask, and the mixture was stirred at room temperature for 4 hours. After the reaction was monitored by LC-MS, the system was concentrated, the residue was added with DCM (20mL), sodium bicarbonate solution (20mL), stirred at room temperature for 30min and then separated, the aqueous phase was extracted with DCM (20mL), and the organic phase was combined with saturated chlorine Wash with sodium chloride solution, dry with anhydrous sodium sulfate, filter, and concentrate to obtain 94-3 (493 mg, yield 96%) as a yellow-brown oil, ESI-MS m/z: 532.3 [M+H] ⁺ .

(S)-2-(cyanomethyl)-4-(7'-(8-methylnaphthalene-1-yl)-1'-(((S)-1-methylpyrrolidin-2-yl )Methyl)-2'-oxo-1',5',7',8'-tetrahydro-2'H-spiro[cyclopropane-1,6'-pyrido[3,4-d]pyrimidine ]-4'-yl)piperazine-1-carboxylic acid benzyl ester (94-4)

Add 94-3 (272mg, 0.512mmol), 1-bromo-8-methylnaphthalene (170mg, 0.768mmol), sodium tert-butoxide (98mg, 1.024mmol), Xantphos (58mg, 0.1mmol) and Dioxane (10 mL), Pd ₂ (dba) ₃ (46 mg, 0.05 mmol) was added after Ar replacement protection, and the mixture was heated to reflux and reacted for 20 h under the protection of Ar. LC-MS monitors after completion of the reaction. The system is quenched by adding water. EA (20mL*2) extraction. The organic phase was washed with saturated sodium chloride. Concentrated and purified by column chromatography (MeOH/DCM=0/1 to 1/40 to 1/20) to obtain a light brown oil 94-4 (62mg, yield 18%), ESI-MS m/z: 672.4[M +H] ⁺ .

2-((S)-4-(7'-(8-methylnaphthalene-1-yl)-1'-(((S)-1-methylpyrrolidin-2-yl)methyl)-2 '-Oxo-1',5',7',8'-tetrahydro-2'H-spiro[cyclopropane-1,6'-pyrido[3,4-d]pyrimidine]-4'-yl )Piperazin-2-yl)acetonitrile (94-5)

Add 94-4 (62 mg, 0.092 mmol), MeOH (10 mL) and 10% Pd/C (20 mg, wet% = 50%) to a 100 mL single-mouth flask, and stir for 20 h at room temperature and normal pressure after H ₂ replacement for three times. After the LC-MS monitoring reaction was completed, the system was filtered, and the filtrate was concentrated to obtain a yellow foamy solid 94-5 (55 mg, yield 100%), ESI-MS m/z: 538.3 [M+H] ⁺ .

2-((S)-1-(2-fluoroacryloyl)-4-(7'-(8-methylnaphthalene-1-yl)-1'-(((S)-1-methylpyrrolidine -2-yl)methyl)-2'-oxy-1',5',7',8'-tetrahydro-2'H-spiro[cyclopropane-1,6'-pyrido[3,4- d)pyrimidine)-4'-yl)piperazin-2-yl)acetonitrile (compound 94)

Add 2-fluoro-acrylic acid (25mg, 0.277mmol), DCM (5mL) and DMF (2mg) to a 50mL single-neck flask. After Ar replacement protection, the temperature is cooled to 0°C, and then oxalyl chloride (30mg, 0.236mmol) in DCM ( 2mL) solution, after dripping, the mixture was stirred at room temperature for 2h. Then the mixture was cooled to 0°C in an ice bath again, and then 94-5 (47 mg, 0.088 mmol) in DCM (2 mL) and DIPEA (57 mg, 0.44 mmol) in DCM (2 mL) were added dropwise in sequence. The mixture was stirred under ice bath for 1 hour. After monitoring the completion of the reaction by LC-MS, the reaction was quenched by adding water (10 mL), and the layers were separated. The aqueous phase was extracted with DCM (10 mL). The combined organic phases were washed with saturated sodium chloride solution (10 mL), dried over anhydrous sodium sulfate, and the crude product was pre -TLC purification gave 94 off-white solid (22mg, yield 41%).

¹ H NMR (400MHz, CDCl ₃ ) δ: 7.64 (d, J = 19.2Hz, 2H), 7.31 (m, 4H), 5.40 (d, J = 47.5 Hz, 1H), 5.23 (d, J = 16.7 Hz ,1H), 4.49(m,1H),4.33-3.97(m,1H),3.74-3.51(m,4H),3.52-3.21(m,5H)3.18-2.75(m,6H),2.65(s, 3H),2.56-2.35(m,4H),1.94-1.82(m,4H),0.62-0.35(m,4H); ESI-MS m/z:610.3[M+H] ⁺ .

Example 95-Example 165: Synthesis of Compound 95-Compound 165

Using intermediates S-1 to S-9 as starting materials, using synthetic method B, similar to the synthetic method of compound 94, compound 95-compound 165 can be obtained.

Example 166: Detection of pERK and ERK protein content in H358 cells by compounds

H358 cells were planted in a 24-well plate. After one day of growth, add the test compound (at a concentration of 1μM). After the compound acts for 24 hours, after lysing the cells, transfer the cell lysate to a 96-well ELISA plate and use an ELISA kit (abcam 176660) Determine the levels of pERK and ERK in the lysate, calculate the ratio of pERK and ERK, and compare it with the DMSO group to calculate the percentage of compounds that inhibit pERK activity. The results are shown in Table 2 below.

Table 2. Inhibitory activity of the compounds of the present invention on pERK levels in H358 cells

Compound	Inhibition rate(%)	Compound	Inhibition rate(%)	Compound	Inhibition rate(%)
1	+++	2	+++	3	+++
4	+++	5	+++	6	+++
7	+++	8	+++	9	+++
10	+++	11	+++	12	+++
13	+++	14	+++	15	+++
16	+++	17	+++	18	+++
19	+++	20	+++	twenty one	++
twenty two	+++	twenty three	+++	twenty four	++

25	+++	26	+++	27	+++
28	+++	29	+++	30	+++
31	+++	32	+++	33	++
34	++	35	++	36	+++
37	+++	38	+++	39	+++
40	+++	41	+++	42	+++
43	++	44	++	45	+++
46	+++	47	+++	48	+++
49	+++	50	+++	51	+++
52	+++	53	+++	54	+++
55	+++	56	+++	57	+++
58	+++	59	+++	60	+++
61	+++	62	+++	63	+++
64	+++	65	+++	66	+++
67	+++	68	+++	69	+++
70	+++	71	+++	72	+++
73	+++	74	+++	75	+++
76	+++	77	+++	78	+++
79	+++	80	+++	81	+++
82	+++	83	+++	84	+++
85	+++	86	+++	87	+++
88	+++	89	+++	90	+++
91	+++	92	+++	93	+++
94	+++	95	+++	96	+++
97	+++	98	++	99	++
100	++	101	++	102	+++
103	+++	104	+++	105	+++
106	++	107	++	108	++
109	++	110	+++	111	+++
112	+++	113	+++	114	++

115	++	116	++	117	++
118	+++	119	+++	120	+++
121	+++	122	++	123	++
124	++	125	++	126	+++
127	+++	128	+++	129	+++
130	++	131	++	132	++
133	++	134	+++	135	+++
136	+++	137	+++	138	++
139	++	140	++	141	++
142	+++	143	+++	144	+++
145	++	146	+++	147	++
148	++	149	++	150	+++
151	+++	152	+++	153	+++
154	++	155	++	156	++
157	++	158	+++	159	+++
160	+++	161	+++	162	++
163	++	164	++	165	++

+ Indicates that the inhibition rate is less than or equal to 50%

++ means the inhibition rate is 50% to 90%

+++ means the inhibition rate is greater than 90%.

Example 167 Anti-proliferative activity of compound on H358 cells

2500 H358 cells were planted in an ultra-low adsorption 96-well plate (corning, 7007). After one day of growth, add gradient dilution compound (up to 30μM, 5-fold dilution, five doses in total), three days after adding the compound, add Cell Titer Glow (Promega, G9681) evaluates the growth of the pellets and calculates the IC ₅₀ value. The results are shown in Table 3 below.

Table 3. Anti-proliferative activity of the compounds of the present invention on H358 cells

Compound	IC 50	Compound	IC 50	Compound	IC 50
1	+++	2	+++	3	+++
4	+++	5	+++	6	+++
7	+++	8	+++	9	+++
10	+++	11	+++	12	+++
13	+++	14	+++	15	+++

16	+++	17	+++	18	+++
19	+++	20	+++	twenty one	++
twenty two	+++	twenty three	+++	twenty four	++
25	+++	26	+++	27	+++
28	+++	29	+++	30	+++
31	+++	32	+++	33	++
34	++	35	++	36	+++
37	+++	38	+++	39	+++
40	+++	41	+++	42	+++
43	++	44	++	45	+++
46	+++	47	+++	48	+++
49	+++	50	+++	51	+++
52	+++	53	+++	54	+++
55	+++	56	+++	57	+++
58	+++	59	+++	60	+++
61	+++	62	+++	63	+++
64	+++	65	+++	66	+++
67	+++	68	+++	69	+++
70	+++	71	+++	72	+++
73	+++	74	+++	75	+++
76	+++	77	+++	78	+++
79	+++	80	+++	81	+++
82	+++	83	+++	84	+++
85	+++	86	+++	87	+++
88	+++	89	+++	90	+++
91	+++	92	+++	93	+++
94	+++	95	+++	96	+++
97	+++	98	++	99	++
100	++	101	++	102	+++
103	+++	104	+++	105	+++

106	++	107	++	108	++
109	++	110	+++	111	+++
112	+++	113	+++	114	++
115	++	116	++	117	++
118	+++	119	+++	120	+++
121	+++	122	++	123	++
124	++	125	++	126	+++
127	+++	128	++	129	++
130	++	131	++	132	+
133	+	134	+++	135	+++
136	+++	137	++	138	++
139	++	140	++	141	+
142	+++	143	+++	144	+++
145	++	146	+++	147	++
148	++	149	++	150	+++
151	+++	152	+++	153	+++
154	++	155	++	156	++
157	++	158	+++	159	+++
160	+++	161	+++	162	++
163	++	164	++	165	++

+ Indicates that the IC _{50 of the} compound is greater than 30 μM

++ means that the IC ₅₀ of the compound is 1 to 30 μM

+++ means that the IC _{50 of the} compound is less than 1 μM.

Although the specific embodiments of the present invention are described above, those skilled in the art should understand that these are merely examples, and various changes or modifications can be made to these embodiments without departing from the principle and essence of the present invention. modify. Therefore, the protection scope of the present invention is defined by the appended claims.

Claims (14)

1. A compound represented by the general formula (1) or its optical isomers, crystal forms, pharmaceutically acceptable salts, hydrates or solvates:

   

   In formula (1):

   L is a chemical bond or NH;

   A is a bivalent 4-12 membered saturated or partially saturated monocyclic, bicyclic, bridged or spiro ring containing 1-2 N atoms. The monocyclic, bicyclic, bridged or spiro ring may be optionally substituted by one R ² is substituted with one or more, when a plurality of R & lt substituted ^2, R ² may be the same or different, R ² is H, CN, C1-C3 alkyl, halogen substituted C1-C3 alkyl or cyano-substituted C1- C3 alkyl;

   R ¹ is an aryl group or a heteroaryl group, and the aryl group or heteroaryl group can be substituted by 1-3 of the following groups: halogen, hydroxyl, amino, C1-C3 alkyl, C2-C4 alkenyl, C3 -C6 cycloalkyl, C1-C3 alkoxy, halogen-substituted C1-C3 alkyl or halogen-substituted C1-C3 alkoxy. When substituted by multiple substituents, the substituents may be the same or different;

   

   Indicates that there is a single bond or double bond between V and Z, when Z is CO and V is

   

   When, there is a single bond between V and Z; or, when V is N, and Z is

   

   When, there is a double bond connection between V and Z;

   R _a , R _b , R _c and R _d are all H, and R _e and R _f and their connected carbon atoms form a C3-C6 cycloalkyl or C4-C6 heterocyclic ring; or, R _c , R _d , R _e and R _f are both H, R _a and R _b and the carbon atom to which they are attached form a C3-C6 cycloalkyl or C4-C6 heterocyclic ring; _{or, R a, R b, R} e and R _f are both H, R _c and R _d and the carbon atom to which they are connected form a C3-C6 cycloalkyl or C4-C6 heterocyclic ring; and

   E is an electrophilic moiety capable of forming a covalent bond with the cysteine residue at position 12 of the K-Ras mutant protein,

   among them,

   Y is a chemical bond or C1-C6 alkylene group;

   R ³ is an amino-substituted alkyl group, cycloalkyl group, alkyl-substituted amido group, heterocyclic group, aryl group or heteroaryl group. The heterocyclic group, aryl group or heteroaryl group can be substituted by 1-3 The group is substituted: halogen, O, CN, OH, hydroxy substituted alkyl, dialkyl substituted amino, C1-C6 alkyl, C3-C6 cycloalkyl, halogen substituted C1-C3 alkyl or halogen substituted C1 -C3 alkoxy, when substituted by multiple substituents, the substituents may be the same or different.
2. The compound according to claim 1, the structure is as shown in general formula (1A) or general formula (1B):

   

   among them:

   L is a chemical bond or NH;

   A is a bivalent 4-12 membered saturated or partially saturated monocyclic, bicyclic, bridged or spiro ring containing 1-2 N atoms. The monocyclic, bicyclic, bridged or spiro ring may be optionally substituted by one R ² is substituted with one or more, when a plurality of R & lt substituted ^2, R ² may be the same or different, R ² is H, CN, C1-C3 alkyl, halogen substituted C1-C3 alkyl or cyano-substituted C1- C3 alkyl;

   R ¹ is an aryl group or a heteroaryl group, and the aryl group or heteroaryl group can be substituted by 1-3 of the following groups: halogen, hydroxyl, amino, C1-C3 alkyl, C2-C4 alkenyl, C3 -C6 cycloalkyl, C1-C3 alkoxy, halogen-substituted C1-C3 alkyl or halogen-substituted C1-C3 alkoxy. When substituted by multiple substituents, the substituents may be the same or different;

   Y is a chemical bond or C1-C6 alkylene group;

   R ³ is an amino-substituted alkyl group, cycloalkyl group, alkyl-substituted amido group, heterocyclic group, aryl group or heteroaryl group. The heterocyclic group, aryl group or heteroaryl group can be substituted by 1-3 The group is substituted: halogen, O, CN, OH, hydroxy substituted alkyl, dialkyl substituted amino, C1-C6 alkyl, C3-C6 cycloalkyl, halogen substituted C1-C3 alkyl or halogen substituted C1 -C3 alkoxy, when it is substituted by multiple substituents, the substituents may be the same or different;

   R _a , R _b , R _c and R _d are all H, and R _e and R _f and their connected carbon atoms form a C3-C6 cycloalkyl or C4-C6 heterocyclic ring; or, R _c , R _d , R _e and R _f are both H, R _a and R _b and the carbon atom to which they are attached form a C3-C6 cycloalkyl or C4-C6 heterocyclic ring; _{or, R a, R b, R} e and R _f are both H, R _c and R _d and the carbon atom to which they are connected form a C3-C6 cycloalkyl or C4-C6 heterocyclic ring; and

   E is an electrophilic moiety capable of forming a covalent bond with the cysteine residue at position 12 of the K-Ras mutant protein.
3. The compound of claim 1 or 2, wherein in the general formula (1), general formula (1A) and general formula (1B), E is a group containing an electrophilic carbon-carbon double bond or a carbon-carbon triple bond group.
4. The compound of claim 3, wherein in the general formula (1), general formula (1A) and general formula (1B), E is:

   

   Among them, R ⁴ is H, F, CF ₃ , OMe or -CH ₂ OMe, and R ⁵ is H, Me, Et, CN, -CONH ₂ , -CH ₂ F, -CHF ₂ , CF ₃ , -CH ₂ OH , CH ₂ OMe,

   

   
5. The compound of claim 1 or 2, wherein in the general formula (1), general formula (1A) and general formula (1B), -ALE is:

   

   

   Wherein, n is 1 or 2, L is a chemical bond or NH, and R ² is H, CN, C1-C3 alkyl, halogen substituted C1-C3 alkyl, or cyano substituted C1-C3 alkyl.
6. The compound according to claim 1 or 2, wherein in the general formula (1), general formula (1A) and general formula (1B), Y is a chemical bond, -CH ₂ -, -CH(Me)- or- CH ₂ CH ₂ -.
7. The compound of claim 1 or 2, wherein in the general formula (1), general formula (1A) and general formula (1B), R ¹ is:

   

   

   Wherein R ⁶ and R ^{7 are} independently H, halogen, hydroxy, amino, C1-C3 alkyl, C2-C4 alkenyl, C3-C6 cycloalkyl, C1-C3 alkoxy, halogen substituted C1-C3 alkyl Or halogen substituted C1-C3 alkoxy.
8. The compound of claim 1 or 2, wherein in the general formula (1), general formula (1A) and general formula (1B), R ³ is:

   

   

   

   Where n is 1, 2 or 3, R ⁸ and R ^{9 are} independently H, halogen, hydroxyl, amino, C1-C3 alkyl, C2-C4 alkenyl, C3-C6 cycloalkyl, C1-C3 alkoxy , Halogen substituted C1-C3 alkyl or halogen substituted C1-C3 alkoxy, R ¹⁰ is C1-C3 alkyl, C3-C6 cycloalkyl, C3-C6 cycloalkyl alkyl, C1-C3 alkoxy alkane Group, halogen substituted C1-C3 alkyl, halogen substituted C3-C6 cycloalkyl or

   
9. The compound of claims 1-8, or a pharmaceutically acceptable salt thereof, wherein the compound has one of the following structures:

   

   

   

   

   

   

   

   
10. A pharmaceutical composition for the treatment, regulation and/or prevention of diseases related to K-Ras G12C mutant protein, characterized in that it contains a pharmaceutically acceptable excipient or carrier, and the right as an active ingredient The compound according to any one of claims 1-9, or each optical isomer, pharmaceutically acceptable salt, hydrate or solvate thereof.
11. The pharmaceutical composition of claim 10, wherein the composition is an oral dosage form.
12. The pharmaceutical composition of claim 10, wherein the composition is in the form of injection.
13. A use of the compound according to any one of claims 1-9, or each of its optical isomers, crystal forms, pharmaceutically acceptable salts, hydrates or solvates, which is characterized in that Individuals in need are methods for diseases mediated by K-Ras G12C mutations.
14. The method of claim 13, wherein the disease is cancer, and the cancer is blood cancer and solid tumor.