WO2022037631A1 - Heterocyclic derivative and preparation method therefor and use thereof - Google Patents

Abstract

The present invention relates to a heterocyclic derivative and a preparation method therefor and use thereof in medicine. Specifically, the present invention relates to a heterocyclic derivative represented by general formula (I), a preparation method therefor and a pharmaceutically acceptable salt thereof, and use thereof as therapeutic agents, especially as K-Ras GTPase inhibitors, wherein the definition of substituents in the general formula (I) is the same as the definition in the description.

Description

Heterocyclic derivatives and preparation methods and uses thereof

This application claims priority to the following Chinese patent applications:

1) The Chinese patent application 202010847639.9 submitted to the Chinese Patent Office on August 21, 2020 with the name of the invention "heterocyclic derivatives and their preparation methods and uses";

2) Chinese patent application No. 202110323635.3 submitted to the Chinese Patent Office on March 26, 2021 with the invention title of "Heterocyclic Derivatives and their Preparation and Use";

3) Chinese patent application 202110569537.X submitted to the Chinese Patent Office on May 25, 2021 with the name of the invention as "Heterocyclic Derivatives and Preparation Methods and Uses thereof";

The contents of each of the above priority applications are incorporated herein by reference in their entirety.

technical field

The present invention relates to a heterocyclic derivative, a preparation method thereof, a pharmaceutical composition containing the derivative and its use as a therapeutic agent, especially as a K-Ras GTPase inhibitor.

Background technique

RAS represents a group of closely related monomeric globular proteins (21 kDa molecular weight) that have 189 amino acids and are associated with the plasma membrane and bind GDP or GTP. Under normal developmental or physiological conditions, RAS is activated by receiving growth factors and various other extracellular signals and is responsible for regulating functions such as cell growth, survival, migration and differentiation. RAS acts as a molecular switch, and the on/off state of RAS proteins is determined by nucleotide binding, with the active signaling conformation bound to GTP and the inactive conformation bound to GDP. When RAS contains bound GDP, it is in a dormant or quiescent or off state and is "inactive". When cells respond to exposure to certain growth-promoting stimuli, RAS is induced and converts bound GDP to GTP. With GTP bound, RAS is "on" and is able to interact with and activate other proteins (its "downstream targets"). The RAS protein itself has a very low intrinsic ability to hydrolyze GTP back to GDP and thereby turn itself into the off state. Switching RAS off requires exogenous proteins called GTPases activating proteins (GAPs), which interact with RAS and greatly facilitate the conversion of GTP to GDP. Any mutation in RAS that affects its ability to interact with GAP or convert GTP back to GDP will result in prolonged activation of the protein and thus an extended signal to the cell that tells it to continue growing and split. So these signals allow cells to grow and divide, and overactive RAS signaling may ultimately lead to cancer.

Structurally, RAS proteins contain a G domain responsible for the enzymatic activity of RAS, guanine nucleotide binding and hydrolysis (GTPase reaction). It also includes a C-terminal extension called the CAAX box, which can be post-translationally modified and target the protein to the membrane. The G domain is approximately 21-25 kDa in size and contains a phosphate-binding loop (P-loop). The P-loop represents the pocket in the protein that binds nucleotides, and this is the rigid part of the domain with conserved amino acid residues necessary for nucleotide binding and hydrolysis (glycine 12, threo amino acid 26 and lysine 16). The G domain also contains the so-called switch I region (residues 30-40) and switch II region (residues 60-76), both of which are the dynamic part of the protein, since the dynamic part has the ability to move between rest and load states The ability to switch is often referred to as a "spring loaded" mechanism. The main interaction is the hydrogen bond formed by threonine-35 and glycine-60 with the γ-phosphate of GTP, which allows switch I and switch II regions, respectively, to maintain their active conformations. After hydrolysis of GTP and release of phosphate, both relax to the inactive GDP conformation.

Among RAS family members, oncogenic mutations were most frequently found in KRAS (85%), whereas NRAS (12%) and HRAS (3%) were less common. KRAS mutations are prevalent in the three most lethal cancer types in the United States: pancreatic cancer (95%), colorectal cancer (45%), and lung cancer (25%). KRAS mutations are also found in other cancer types including carcinoma, diffuse large B-cell lymphoma, rhabdomyosarcoma, cutaneous squamous cell carcinoma, cervical cancer, testicular germ cell carcinoma, etc. Rarely found (<2%). In non-small cell lung cancer (NSCLC), KRAS G12C is the most common mutation, accounting for nearly half of all KRAS mutations, followed by G12V and G12D. In non-small cell lung cancer, the increased frequency of allele-specific mutations is mostly due to classic smoking-induced mutations (G:C to T:A substitutions), resulting in KRAS G12C (GGT to TGT) and G12V (GGT) to GTT) mutation.

Large genomic studies have shown that KRAS mutations in lung cancer, including G12C, are mutually exclusive with other known driver oncogenic mutations in NSCLC, including EGFR, ALK, ROS1, RET, and BRAF, suggesting the uniqueness of KRAS mutations in lung cancer. Meanwhile, KRAS mutations frequently co-occur with certain co-mutations, such as STK11, KEAP1, and TP53, which cooperate with mutated RAS to transform cells into highly malignant and aggressive tumor cells.

Three RAS oncogenes constitute the most frequently mutated gene family in human cancers. Disappointingly, despite more than three decades of research efforts, there is still no clinically effective anti-RAS therapy, and targeting this gene with small molecules is challenging. Therefore, there is an urgent need in the art for small molecules for targeting RAS (eg, K-RAS, H-RAS and/or N-RAS) and using them to treat various diseases, such as cancer.

At present, there is fierce competition for clinical development of KRAS inhibitors at home and abroad. Among them, MRTX-849, a KRAS enzyme inhibitor developed by Mirati Therapeutics Inc, has entered clinical phase II for the prevention and treatment of advanced solid tumors, metastatic colorectal cancer and metastasis diseases such as non-small cell lung cancer. There are also other KRAS inhibitors under investigation, including AMG-510 (Amgen Inc, phase 2) and MRTX1257 (Mirati Therapeutics Inc, Discovery). Early clinical studies have shown that KRAS inhibitors can control and alleviate disease progression in patients with non-small cell lung cancer and can reduce tumor size in patients with advanced lung and colorectal cancer. At present, a series of KRAS inhibitor patent applications have been published, including WO2020047192, WO2019099524 and WO2018217651, etc., indicating that the research and application of KRAS inhibitors have made certain progress. However, the existing KRAS inhibitors are not effective and safe. The aspects are still not satisfactory, the room for improvement is still huge, and it is still necessary to continue the research and development of new KRAS inhibitors.

SUMMARY OF THE INVENTION

The present inventors unexpectedly found in research that the tetracyclic derivatives represented by the following general formula (I), or stereoisomers, tautomers or pharmaceutically acceptable salts thereof can be used as effective KRAS inhibitors agent with good efficacy and safety.

Based on the above findings, in the first aspect, the present invention provides a heterocyclic derivative represented by the general formula (I), or a stereoisomer, tautomer or a pharmaceutically acceptable salt thereof:

in:

E is selected from

G is a 4- to 12-membered heterocyclic group containing 1-2 nitrogen atoms, preferably a 4-membered heterocyclic group, wherein the heterocyclic group is optionally further substituted by one or more R ^c ;

L is a chemical bond or a C ₁ -C ₆ alkylene group; wherein the alkylene group is optionally further substituted by one or more substituents selected from alkyl, halogen or hydroxyl; preferably, L is a chemical bond, -CH ₂ -, -CH ₂ CH ₂ - or -CH(CH ₃ )-; more preferably, L is a chemical bond;

X and Y are each independently selected from N or CR ^f ;

Ring A is selected from the group:

The condition is that when ring A is selected from

When , X is selected from N, Y is selected from CR ^f , the carbon atom of G is connected with ring A;

When ring A is selected from

, -LR ⁴ does not exist;

W is selected from N or CR ^d ;

Z is selected from N or CR ^e ;

Ring B is selected from 5-6 membered heteroaryl;

Ring C is selected from 5-6 membered heteroaryl;

^Ra is selected from hydrogen atoms or fluorine;

R ^b is selected from hydrogen atom, -CH ₂ F, -CHF ₂ ,

R ^c is the same or different, and each is independently selected from a hydrogen atom, an alkyl group, a halogen, a nitro group, a cyano group, a cycloalkyl group, a heterocyclic group, an aryl group, a heteroaryl group, =O, -OR ⁵ , -C( O)R ⁵ , -C(O)OR ⁵ , -NHC(O)R ⁵ , -NHC(O)OR ⁵ , -NR ⁶ R ⁷ , -C(O)NR ⁶ R ⁷ , -CH ₂ NHC ( O)OR ⁵ , -CH ₂ NR ⁶ R ⁷ or -S(O) _r R ⁵ , wherein said alkyl, cycloalkyl, heterocyclyl, aryl or heteroaryl is optionally further modified by one or more Multiple selected from alkyl, halogen, nitro, cyano, cycloalkyl, heterocyclyl, aryl, heteroaryl, =O, ^-OR5 , -C(O) ^R5 , -C(O) OR ⁵ , -NHC(O)R ⁵ , -NHC(O)OR ⁵ , -NR ⁶ R ⁷ , -C(O)NR ⁶ R ⁷ , -CH ₂ NHC(O)OR ⁵ , -CH ₂ NR ⁶ R ⁷ or -S(O) _r R ⁵ is substituted by the substituent;

R ^d and R ^e are the same or different, each independently selected from hydrogen atom, halogen, alkyl, alkoxy, cyano, nitro, amino, hydroxyl, haloalkyl or haloalkoxy, preferably cyano;

R ^{f is} selected from hydrogen atom, halogen, alkyl or alkoxy; wherein said alkyl or alkoxy is optionally further selected by one or more of halogen, hydroxy, cyano, alkyl or alkoxy substituted by the substituent; R ^f is preferably halogen, more preferably fluorine or chlorine;

R ¹ is selected from hydrogen atom, halogen, alkyl or alkoxy; wherein said alkyl or alkoxy is optionally further selected by one or more of halogen, hydroxyl, cyano, alkyl or alkoxy substituted by the substituent; R ¹ is preferably a hydrogen atom;

R ² is selected from aryl or heteroaryl, wherein said aryl or heteroaryl is optionally further substituted with one or more ^RA ;

R ^A is each independently selected from alkyl, halogen, nitro, cyano, cycloalkyl, heterocyclyl, aryl, heteroaryl, =O, ^-OR5 , -C(O) ^R5 , -C (O)OR ⁵ , -NHC(O)R ⁵ -NHC(O)OR ⁵ , -NR ⁶ R ⁷ , -C(O)NR ⁶ R ⁷ , -CH ₂ NHC(O)OR ⁵ , -CH ₂ NR ⁶ R ⁷ or -S(O) _r R ⁵ ; wherein said alkyl, cycloalkyl, heterocyclyl, aryl or heteroaryl is optionally further , nitro, cyano, cycloalkyl, heterocyclyl, aryl, heteroaryl, =O, -OR ⁵ , -C(O)R ⁵ , -C(O)OR ⁵ , -NHC(O) R ⁵ -NHC(O)OR ⁵ , -NR ⁶ R ⁷ , -C(O)NR ⁶ R ⁷ , -CH ₂ NHC(O)OR ⁵ , -CH ₂ NR ⁶ R ⁷ or -S(O) _r Replaced by the substituent of R ⁵ ;

The condition is that when ring A is selected from

, R ² is not selected from

R ³ is selected from hydrogen atom, halogen, alkyl, alkoxy, cyano, haloalkyl or haloalkoxy, preferably hydrogen;

R ⁴ is selected from a hydrogen atom, an aryl group or a heteroaryl group; wherein said aryl or heteroaryl group is optionally further substituted by one or more R ^B ; R ⁴ is preferably a heteroaryl group;

Each R ^B is independently selected from alkyl, halogen, nitro, cyano, cycloalkyl, heterocyclyl, aryl, heteroaryl, =O, ^-OR5 , -C(O) ^R5 , -C (O)OR ⁵ , -NHC(O)R ⁵ -NHC(O)OR ⁵ , -NR ⁶ R ⁷ , -C(O)NR ⁶ R ⁷ , -CH ₂ NHC(O)OR ⁵ , -CH ₂ NR ⁶ R ⁷ or -S(O) _r R ⁵ ; wherein said alkyl, cycloalkyl, heterocyclyl, aryl or heteroaryl is optionally further , nitro, cyano, cycloalkyl, heterocyclyl, aryl, heteroaryl, =O, -OR ⁵ , -C(O)R ⁵ , -C(O)OR ⁵ , -NHC(O) R ⁵ -NHC(O)OR ⁵ , -NR ⁶ R ⁷ , -C(O)NR ⁶ R ⁷ , -CH ₂ NHC(O)OR ⁵ , -CH ₂ NR ⁶ R ⁷ or -S(O) _r Replaced by the substituent of R ⁵ ;

R ⁵ is each independently selected from a hydrogen atom, an alkyl group, a cycloalkyl group, a heterocyclyl group, an aryl group or a heteroaryl group, wherein said alkyl group, cycloalkyl group, heterocyclyl group, aryl group or heteroaryl group is any is further selected by one or more selected from the group consisting of hydroxy, halogen, nitro, cyano, alkyl, alkoxy, haloalkyl, haloalkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl, = O, -C(O) ^R8 , _-C (O) ^OR8 , ^-OC (O) ^R8 , ^-NR9R10 , ^-C ( ^O ) ^NR9R10 , ^-SO2NR9R10 or -Replaced by the substituent of NR ⁹ C(O)R ¹⁰ ;

R ⁶ and R ⁷ are each independently selected from a hydrogen atom, a hydroxyl group, a halogen, an alkyl group, an alkoxy group, a cycloalkyl group, a heterocyclic group, an aryl group or a heteroaryl group, wherein the alkyl group, alkoxy group, A cycloalkyl, heterocyclyl, aryl or heteroaryl group is optionally further selected from one or more groups selected from hydroxy, halogen, nitro, cyano, alkyl, alkoxy, cycloalkyl, heterocyclyl, Aryl, heteroaryl, =O, -C(O)R ⁸ , -C(O)OR ⁸ , -OC(O)R ⁸ , -NR ⁹ R ¹⁰ , -C(O)NR ⁹ R ¹⁰ , -SO ₂ NR ⁹ R ¹⁰ or -NR ⁹ C(O)R ¹⁰ substituent;

Alternatively, R ⁶ and R ⁷ together with the atoms to which they are attached form a 4- to 8-membered heterocyclyl group containing one or more N, O or S(O) _r within the 4- to 8-membered heterocyclyl group, and The 4-8 membered heterocyclic group is optionally further selected from one or more groups selected from hydroxyl, halogen, nitro, cyano, alkyl, alkoxy, cycloalkyl, heterocyclic, aryl, heterocyclic Aryl, =O, -C(O)R ⁸ , -C(O)OR ⁸ , -OC(O)R ⁸ , -NR ⁹ R ¹⁰ , -C(O)NR ⁹ R ¹⁰ , -SO ₂ NR ⁹ R ¹⁰ or -NR ⁹ C(O) R ¹⁰ is substituted with a substituent;

R ⁸ , R ⁹ and R ¹⁰ are each independently selected from a hydrogen atom, an alkyl group, an amino group, a cycloalkyl group, a heterocyclic group, an aryl group or a heteroaryl group, wherein said alkyl group, cycloalkyl group, heterocyclic group , aryl or heteroaryl are optionally further selected from one or more groups selected from hydroxy, halogen, nitro, amino, cyano, alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl substituted by substituents of carboxylate, carboxyl or carboxylate;

R ¹¹ is selected from hydrogen atom, halogen, alkyl, alkoxy, cyano, nitro, amino, hydroxyl, haloalkyl, haloalkoxy or =O;

R ¹² is selected from a hydrogen atom, an alkyl group, -C(O)R ¹³ or -S(O) ₂ R ¹³ ;

R ¹³ is selected from alkyl, preferably methyl;

n is each independently selected from 0, 1 or 2;

r is each independently 0, 1 or 2.

In a preferred embodiment of the present invention, the compound represented by the general formula (I) or its stereoisomer, tautomer or its pharmaceutically acceptable salt is the compound represented by the general formula (II) or its Stereoisomers, tautomers or pharmaceutically acceptable salts thereof:

in:

Ring B, R ¹ to R ³ , X, Y, G, E and n are as defined in the general formula (I).

In a preferred embodiment of the present invention, for the compound of general formula (I) or (II) or its stereoisomer, tautomer or its pharmaceutically acceptable salt, wherein:

-G-E are selected from:

R ^c are the same or different, each independently selected from hydrogen atom, halogen, alkyl or alkoxy, preferably alkyl, more preferably methyl;

m is selected from 0, 1, 2, 3 or 4;

E is as defined in general formula (I).

In a preferred embodiment of the present invention, for the compound of general formula (I) or (II) or its stereoisomer, tautomer or its pharmaceutically acceptable salt, wherein E is selected from:

In a preferred embodiment of the present invention, for the compound of general formula (I) or (II) or its stereoisomer, tautomer or its pharmaceutically acceptable salt, wherein X and Y are each independently is selected from CR ^f ; R ^{f is} selected from hydrogen atom, halogen, alkyl, alkoxy, haloalkyl or haloalkoxy, R ^f is more preferably halogen, particularly preferably fluorine or chlorine.

In a preferred embodiment of the present invention, for the compound of general formula (I) or (II) or its stereoisomer, tautomer or its pharmaceutically acceptable salt, wherein R ¹ is selected from hydrogen atom, halogen, alkyl, alkoxy, haloalkyl or haloalkoxy, R ¹ is more preferably a hydrogen atom.

In a preferred embodiment of the present invention, for the compound of general formula (I) or (II) or its stereoisomer, tautomer or its pharmaceutically acceptable salt, wherein:

R ² is selected from phenyl, naphthyl, pyridyl, benzothiazolyl or benzopyrazolyl, wherein said phenyl, naphthyl, pyridyl, benzothiazolyl or benzopyrazolyl is optionally further replaced by one or more ^RAs ;

R ^A is each independently selected from halogen, hydroxy, alkyl, alkoxy, cycloalkyl or -NR ⁶ R ⁷ , wherein said alkyl or alkoxy is optionally further selected by one or more selected from halogen or -Substituents of NR ⁶ R ⁷ are substituted; wherein the halogen is preferably fluorine;

R ⁶ and R ⁷ are each independently selected from a hydrogen atom or an alkyl group, wherein the alkyl group is more preferably a methyl group.

In a preferred embodiment of the present invention, for the compound of general formula (I) or (II) or its stereoisomer, tautomer or its pharmaceutically acceptable salt, wherein R ² is selected from:

In a preferred embodiment of the present invention, for the compound of general formula (II) or its stereoisomer, tautomer or its pharmaceutically acceptable salt, wherein R ³ is selected from hydrogen atom.

In a preferred embodiment of the present invention, for the compound of general formula (II) or its stereoisomer, tautomer or its pharmaceutically acceptable salt, wherein ring B is selected from:

Typical compounds of the present invention include, but are not limited to:

or its stereoisomers, tautomers or pharmaceutically acceptable salts thereof.

Note: If there is a discrepancy between the structural formula and the given name of the structural formula, the structural formula shall prevail.

Further, the present invention provides a preparation method of the compound of general formula (II) or its stereoisomer, tautomer or pharmaceutically acceptable salt thereof, said method comprising the following steps:

The compound of general formula (IIA) is reacted with the compound of general formula (IIB) under basic conditions, and optionally the protective group is further removed to obtain the compound of general formula (II);

in:

X ¹ is a leaving group, and the leaving group is preferably chlorine, bromine, iodine, OMs or OTs; more preferably chlorine;

Ring B, R ¹ to R ³ , X, Y, G, E and n are as defined in the general formula (II).

In another aspect, the present invention provides a pharmaceutical composition containing an effective dose of the compound of general formula (I) or (II) or a stereoisomer, tautomer or A pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, excipient or a combination thereof.

In another aspect, the present invention provides a method for inhibiting K-Ras GTPase, wherein the method comprises, administering to a subject (including patients and healthy subjects) in need thereof a pharmaceutical composition, the medicine The composition contains an effective dose of the compound described in general formula (I) or (II) or its stereoisomer, tautomer or its pharmaceutically acceptable salt, and optional pharmaceutically acceptable carrier, excipient. Form or their combination, wherein K-Ras GTPase is preferably KRAS G12C.

The present invention also provides a compound of the general formula (I) or (II) or a stereoisomer, a tautomer or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof prepared for treatment Use in medicine for diseases mediated by KRAS mutation, wherein said disease mediated by KRAS mutation is preferably selected from cancer, wherein said cancer is preferably selected from pancreatic cancer, colorectal cancer, lung cancer, multiple myeloma , uterine cancer, bile duct cancer, gastric cancer, bladder cancer, diffuse large B-cell lymphoma, rhabdomyosarcoma, skin squamous cell carcinoma, cervical cancer, testicular germ cell cancer, more preferably pancreatic cancer, colorectal cancer and lung cancer; wherein The lung cancer is preferably non-small cell lung cancer.

In another aspect, the present invention provides a compound of the general formula (I) or (II) or a stereoisomer, a tautomer or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof in the preparation Use in a K-Ras GTPase inhibitor, wherein the K-Ras GTPase inhibitor is preferably a KRAS G12C inhibitor.

Another aspect of the present invention relates to a method of preventing and/or treating KRAS mutation-mediated diseases, comprising administering to a patient a therapeutically effective dose of a compound represented by general formula (I) or (II) or a tautomer thereof isomer, meso, racemate, enantiomer, diastereomer or a mixture thereof or a pharmaceutically acceptable salt thereof or a pharmaceutical composition comprising the same, wherein the KRAS mutation is preferably for the KRAS G12C mutation.

The present invention also provides a compound of the general formula (I) or (II) or a stereoisomer, a tautomer or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof prepared for treatment Use in the medicament of cancer, wherein said cancer is preferably selected from pancreatic cancer, colorectal cancer, lung cancer, multiple myeloma, uterine cancer, bile duct cancer, gastric cancer, bladder cancer, diffuse large B-cell lymphoma, rhabdomyosarcoma , skin squamous cell carcinoma, cervical cancer, testicular germ cell carcinoma, more preferably pancreatic cancer, colorectal cancer and lung cancer; wherein the lung cancer is preferably non-small cell lung cancer.

The pharmaceutical formulations of the present invention can be topical, oral, transdermal, rectal, vaginal, parenteral, intranasal, intrapulmonary, intraocular, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intradermal , intraperitoneal, subcutaneous, substratum corneum, or by inhalation. Pharmaceutical compositions containing the active ingredient may be in a form suitable for oral administration, such as tablets, dragees, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsions, hard or soft capsules, or syrups or Elixirs. Tablets may contain the active ingredient in admixture with suitable nontoxic pharmaceutically acceptable excipients for the manufacture of tablets.

The formulations of the present invention are suitably presented in unit dosage form, and such formulations may be prepared by any of the methods well known in the art of pharmacy. The amount of active ingredient that can be combined with carrier materials to produce a single dosage form can vary depending upon the host treated and the particular mode of administration. The amount of active ingredient which, in combination with a carrier material, can produce a single dosage form generally refers to that amount of compound which produces a therapeutic effect.

Dosage forms for topical or transdermal administration of the compounds of this invention may include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches and inhalants. The active compound can be mixed under sterile conditions with a pharmaceutically acceptable carrier, and it can be mixed with any preservatives, buffers or propellants that may be required.

When the compounds of the present invention are administered in pharmaceutical form to humans and animals, the compounds may be provided alone or in the form of pharmaceutical compositions, which may contain admixture with a pharmaceutically acceptable carrier. Active ingredients in combination, eg, 0.1% to 99.5% (more preferably, 0.5% to 90%) active ingredients.

Examples of pharmaceutically acceptable carriers include, but are not limited to: (1) sugars such as lactose, glucose and sucrose; (2) starches such as corn starch and potato starch; (3) cellulose and derivatives thereof such as carboxylate (4) powdered gum tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients such as cocoa butter and Suppository waxes; (9) oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; (10) glycols such as propylene glycol; (11) polyols such as glycerol, sorbitol , mannitol and polyethylene glycol; (12) esters such as ethyl oleate and ethyl laurate; (13) agar; (14) buffers such as magnesium hydroxide and aluminum hydroxide; (15) seaweed (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) ethanol; (20) phosphate buffer solution; (21) cyclodextrin, e.g. linked Targeting ligands in nanoparticles, such as Accurins™; and (22) other non-toxic compatible substances in pharmaceutical formulations, such as polymer-based compositions.

Examples of pharmaceutically acceptable antioxidants include, but are not limited to: (1) water-soluble antioxidants such as ascorbic acid, cysteine hydrochloride, sodium bisulfite, sodium metabisulfite, sodium sulfite, and the like; ( 2) Oil-soluble antioxidants such as ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, alpha-tocopherol and the like; and (3) metal chelating agents such as citric acid, ethylenediaminetetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid and the like. Solid dosage forms (eg, capsules, dragees, dragees, powders, granules, and the like) may include one or more pharmaceutically acceptable carriers, such as sodium citrate or dicalcium phosphate, and/or the following Any one of: (1) fillers or bulking agents, such as starch, lactose, sucrose, glucose, mannitol and/or silicic acid; (2) binders, such as carboxymethyl cellulose, alginate, Gelatin, polyvinylpyrrolidone, sucrose and/or acacia; (3) humectants, such as glycerol; (4) disintegrants, such as agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates and sodium carbonate; (5) dissolution retarders, such as paraffin; (6) absorption accelerators, such as quaternary ammonium compounds; (7) humectants, such as cetyl alcohol and glycerol monostearate; (8) absorption agents such as kaolin and bentonite; (9) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate and mixtures thereof; and (10) colorants. Liquid dosage forms can include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active ingredient, liquid dosage forms may contain inert diluents commonly used in the art, such as water or other solvents; solubilizers and emulsifiers, such as ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzene Methanol, benzyl benzoate, propylene glycol, 1,3-butanediol, oils (especially cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil, and sesame oil), glycerin, tetrahydrofuran methanol, polyethylene Diols and fatty acid esters of sorbitan, and mixtures thereof.

Suspensions, in addition to the active compounds, may also contain suspending agents such as ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum hydroxide oxide, bentonite, Agar and tragacanth and mixtures thereof.

Ointments, pastes, creams and gels can contain, in addition to the active compound, excipients such as animal and vegetable fats, oils, waxes, paraffins, starches, tragacanth, cellulose derivatives, poly Ethylene glycol, polysiloxane, bentonite, silicic acid, talc and zinc oxide or mixtures thereof.

Powders and sprays can contain, in addition to the active compound, excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicates and polyamide powder, or mixtures of these substances. The sprays can contain other customary propellants, such as chlorofluorohydrocarbons, and volatile unsubstituted hydrocarbons, such as butane and propane.

Detailed description of the invention

Unless stated to the contrary, some terms used in the specification and claims of the present invention are defined as follows:

"Alkyl" when taken as a group or part of a group is meant to include _C1 - _C20 straight or branched chain aliphatic hydrocarbon groups. Preferably it is a C ₁ -C ₁₀ alkyl group, more preferably a C ₁ -C ₆ alkyl group, or a C ₁ -C ₄ alkyl group. Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-di Methylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1 -Ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethyl Butyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl Wait. Alkyl groups can be substituted or unsubstituted.

"Alkenyl" refers to an alkyl group as defined above consisting of at least two carbon atoms and at least one carbon-carbon double bond, representative examples include, but are not limited to, ethenyl, 1-propenyl, 2-propenyl, 1-, 2- or 3-butenyl, etc. Preferred are C2 _{- C10} alkenyl groups, more preferably C2 _{- C6} alkenyl groups, and most preferably C2 _{- C4} alkenyl groups. Alkenyl groups can be optionally substituted or unsubstituted.

"Alkynyl" refers to an aliphatic hydrocarbon group containing a carbon-carbon triple bond, which may be straight or branched. Preferred are C2 _{- C10} alkynyl groups, more preferably C2 _{- C6} alkynyl groups, and most preferably C2 _{- C4} alkynyl groups. Examples of alkynyl groups include, but are not limited to, ethynyl, 1-propynyl, 2-propynyl, 1-, 2- or 3-butynyl, and the like. Alkynyl groups can be substituted or unsubstituted.

"Cycloalkyl" refers to saturated or partially saturated monocyclic, fused, bridged and spirocyclic carbocyclic rings. Preferably it is C ₃ -C ₁₂ cycloalkyl, more preferably C ₃ -C ₈ cycloalkyl, most preferably C ₃ -C ₆ cycloalkyl. Examples of monocyclic cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptyl Alkenyl, cyclooctyl, etc., preferably cyclopropyl and cyclohexenyl. Cycloalkyl groups can be optionally substituted or unsubstituted.

"Spirocycloalkyl" refers to a polycyclic group with 5 to 18 members, two or more cyclic structures, and the single rings share one carbon atom (called spiro atom) with each other, and the ring contains one or more aromatic systems with double bonds but none of the rings have fully conjugated pi electrons. The spirocycloalkyl group is preferably a 6- to 14-membered spirocycloalkyl group, more preferably a 7- to 10-membered spirocycloalkyl group. According to the number of spiro atoms shared between the rings, spirocycloalkyl groups are divided into mono-spiro, double-spiro or poly-spirocycloalkyl groups, preferably mono-spiro and double-spirocycloalkyl groups, preferably 4-membered/5-membered, 4-membered Yuan/6 Yuan, 5 Yuan/5 Yuan or 5 Yuan/6 Yuan. Non-limiting examples of "spirocycloalkyl" include, but are not limited to, spiro[4.5]decyl, spiro[4.4]nonyl, spiro[3.5]nonyl, spiro[2.4]heptyl.

"Fused cycloalkyl" refers to a 5- to 18-membered all-carbon polycyclic group containing two or more cyclic structures sharing a pair of carbon atoms with each other, one or more rings may contain one or more double bonds, An aromatic system in which none of the rings has a fully conjugated pi electron, preferably a 6- to 12-membered fused cycloalkyl group, more preferably a 7- to 10-membered fused cycloalkyl group. According to the number of constituent rings, it can be divided into bicyclic, tricyclic, tetracyclic or polycyclic fused cycloalkyl, preferably bicyclic or tricyclic, more preferably 5-membered/5-membered or 5-membered/6-membered bicycloalkyl. Non-limiting examples of "fused cycloalkyl" include, but are not limited to: bicyclo[3.1.0]hexyl, bicyclo[3.2.0]hept-1-enyl, bicyclo[3.2.0]heptyl, Decalinyl or tetrahydrophenanthryl.

"Bridged cycloalkyl" refers to an all-carbon polycyclic group of 5 to 18 members, containing two or more cyclic structures, sharing two carbon atoms that are not directly connected to each other, and one or more rings may contain one or more Aromatic systems in which multiple double bonds, but none of the rings have fully conjugated pi electrons, are preferably 6- to 12-membered bridged cycloalkyl, more preferably 7- to 10-membered bridged cycloalkyl. According to the number of constituent rings, it can be divided into bicyclic, tricyclic, tetracyclic or polycyclic bridged cycloalkyl, preferably bicyclic, tricyclic or tetracyclic, more preferably bicyclic or tricyclic. Non-limiting examples of "bridged cycloalkyl" include, but are not limited to: (1s,4s)-bicyclo[2.2.1]heptyl, bicyclo[3.2.1]octyl, (1s,5s)-dicyclo[3.2.1]octyl Cyclo[3.3.1]nonyl, bicyclo[2.2.2]octyl, (1r,5r)-bicyclo[3.3.2]decyl.

"Heterocyclyl," "heterocycle," or "heterocyclic" are used interchangeably herein to refer to a non-aromatic heterocyclyl in which one or more of the ring-forming atoms is a heteroatom, such as oxygen, Nitrogen, sulfur atoms, etc., including monocyclic, fused, bridged and spiro rings. It preferably has a 5- to 7-membered monocyclic ring or a 7- to 10-membered bi- or tricyclic ring, which may contain 1, 2 or 3 atoms selected from nitrogen, oxygen and/or sulfur. Examples of "heterocyclyl" include, but are not limited to, morpholinyl, oxetanyl, thiomorpholinyl, tetrahydropyranyl, 1,1-dioxothiomorpholinyl, piperidinyl , 2-oxopiperidinyl, pyrrolidinyl, 2-oxopyrrolidinyl, piperazin-2-one, 8-oxa-3-aza-bicyclo[3.2.1]octyl and piperazinyl . Heterocyclyl groups can be substituted or unsubstituted.

"Spiroheterocyclyl" refers to a polycyclic group with 5 to 18 members, two or more cyclic structures, and single rings share one atom with each other, and the ring contains one or more double bonds, but no An aromatic system with fully conjugated pi electrons in one ring, wherein one or more ring atoms are selected from nitrogen, oxygen, or a heteroatom of S(O) _r (wherein r is selected from 0, 1, or 2), and the remaining ring atoms are carbon. It is preferably a 6- to 14-membered spiroheterocyclic group, more preferably a 7- to 10-membered spiroheterocyclic group. Spirocycloalkyl groups are classified into mono-spiroheterocyclyl, bis-spiroheterocyclyl or poly-spiroheterocyclyl according to the number of spiro atoms shared between rings, preferably mono-spiroheterocyclyl and bis-spiroheterocyclyl. More preferably, it is a 4-membered/4-membered, 4-membered/5-membered, 4-membered/6-membered, 5-membered/5-membered or 5-membered/6-membered monospiroheterocyclyl group. Non-limiting examples of "spiroheterocyclyl" include, but are not limited to: 1,7-dioxaspiro[4.5]decyl, 2-oxa-7-azaspiro[4.4]nonyl, 7-oxaspiro[4.4]nonyl Heterospiro[3.5]nonyl and 5-oxaspiro[2.4]heptyl.

"Fused heterocyclic group" refers to an all-carbon polycyclic group containing two or more ring structures sharing a pair of atoms with each other, one or more rings may contain one or more double bonds, but no ring has complete Conjugated pi-electron aromatic systems wherein one or more ring atoms are selected from nitrogen, oxygen or heteroatoms of S(O) _r (wherein r is selected from 0, 1 or 2) and the remaining ring atoms are carbon. It is preferably a 6- to 14-membered condensed heterocyclic group, more preferably a 7- to 10-membered condensed heterocyclic group. According to the number of constituent rings, it can be divided into bicyclic, tricyclic, tetracyclic or polycyclic fused heterocyclic groups, preferably bicyclic or tricyclic, more preferably 5-membered/5-membered or 5-membered/6-membered bicyclic fused heterocyclic groups. Non-limiting examples of "fused heterocyclyl" include, but are not limited to: octahydropyrrolo[3,4-c]pyrrolyl, octahydro-1H-isoindolyl, 3-azabicyclo[3.1. 0] Hexyl, octahydrobenzo[b][1,4]dioxine.

"Bridged heterocyclyl" refers to a 5- to 14-membered, 5- to 18-membered polycyclic group containing two or more cyclic structures that share two atoms that are not directly connected to each other, and one or more rings may be Aromatic systems containing one or more double bonds but none of the rings have fully conjugated pi electrons, wherein one or more ring atoms are selected from nitrogen, oxygen or S(O) _r (wherein r is selected from 0, 1 or 2), the remaining ring atoms are carbon. Preferably it is a 6- to 14-membered bridged heterocyclic group, more preferably a 7- to 10-membered bridged heterocyclic group. According to the number of constituent rings, it can be divided into bicyclic, tricyclic, tetracyclic or polycyclic bridged heterocyclic groups, preferably bicyclic, tricyclic or tetracyclic, more preferably bicyclic or tricyclic. Non-limiting examples of "bridged heterocyclyl" include, but are not limited to: 2-azabicyclo[2.2.1]heptyl, 2-azabicyclo[2.2.2]octyl, and 2-azabicyclo Cyclo[3.3.2]decyl.

"Aryl" refers to a carbocyclic aromatic system containing one or two rings, wherein the rings may be joined together in a fused fashion. The term "aryl" includes monocyclic or bicyclic aryl groups such as phenyl, naphthyl, tetrahydronaphthyl aromatic groups. Preferred aryl groups are _C6 - _C10 aryl groups, more preferred aryl groups are phenyl and naphthyl. Aryl groups can be substituted or unsubstituted.

"Heteroaryl" refers to an aromatic 5 to 6 membered monocyclic or 8 to 10 membered bicyclic ring, which may contain 1 to 4 atoms selected from nitrogen, oxygen and/or sulfur. Preferably bicyclic heteroaryl, examples of "heteroaryl" include but are not limited to the following: furyl, pyridyl, 2-oxo-1,2-dihydropyridyl, pyridazinyl, pyrimidinyl, pyridyl Azinyl, thienyl, isoxazolyl, oxazolyl, oxadiazolyl, imidazolyl, pyrrolyl, pyrazolyl, triazolyl, tetrazolyl, thiazolyl, isothiazolyl, 1,2, 3-thiadiazolyl, benzodioxolyl, benzothienyl, benzimidazolyl, indolyl, isoindolyl, 1,3-dioxo-isoindolyl, Quinolinyl, Indazolyl, Benzisothiazolyl, Benzoxazolyl, Benzisoxazolyl,

Heteroaryl groups in the present invention may be substituted or unsubstituted.

"Alkoxy" refers to a group (alkyl-O-). Wherein, alkyl is as defined herein. C ₁ -C ₆ and C ₁ -C ₄ alkoxy groups are preferred. Examples include, but are not limited to: methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, and the like.

"Haloalkyl" means a group in which an alkyl group is optionally further substituted with one or more halogens, wherein alkyl is as defined herein.

"Hydroxyalkyl" refers to a group in which an alkyl group is optionally further substituted with one or more hydroxy groups, wherein alkyl is as defined herein.

"Haloalkoxy" refers to a group in which the alkyl group of (alkyl-O-) is optionally further substituted with one or more halogens, wherein alkoxy is as defined herein.

"Hydroxy" refers to the -OH group.

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

"Amino" refers to _-NH2 .

"Cyano" refers to -CN.

"Nitro" refers to _-NO2 .

"Benzyl" refers to _-CH2 -phenyl.

"Carboxyl" refers to -C(O)OH.

"Carboxylate" means -C(O)O-alkyl or -C(O)O-cycloalkyl, wherein alkyl and cycloalkyl are as defined above.

"DMSO" refers to dimethyl sulfoxide.

"BOC" refers to t-butoxycarbonyl.

"Ts" refers to p-toluenesulfonyl.

"Ms" methylsulfonyl.

"T3P" refers to propylphosphoric anhydride.

"DPPA" refers to diphenylphosphoryl azide.

"DEA" refers to diethylamine.

"X-PHOS Pd G2" Chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1 ,1'-biphenyl)]palladium(II).

"Substituted" means that one or more hydrogen atoms in a group, preferably up to 5, more preferably 1 to 3 hydrogen atoms, independently of one another, are substituted with a corresponding number of substituents. It goes without saying that the substituents are only in their possible chemical positions, and the person skilled in the art can determine (either experimentally or theoretically) possible or impossible substitutions without undue effort. For example, amino or hydroxyl groups with free hydrogens may be unstable when combined with carbon atoms with unsaturated (eg, olefinic) bonds.

"Substituted" or "substituted" in this specification, unless otherwise specified, means that the group may be substituted by one or more groups selected from the group consisting of: alkyl, alkenyl, alkynyl, alkoxy group, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocyclyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, ring Alkylthio, heterocycloalkylthio, amino, haloalkyl, hydroxyalkyl, carboxyl, carboxylate, =O, -C(O)R ⁵ , -C(O)OR ⁵ , -NHC(O) R ⁵ , -NHC(O)OR ⁵ , -NR ⁶ R ⁷ , -C(O)NR ⁶ R ⁷ , -CH ₂ NHC(O)OR ⁵ , -CH ₂ NR ⁶ R ⁷ or -S(O) _r R ⁵ ;

Wherein, R ⁵ is selected from hydrogen atom, alkyl group, cycloalkyl group, heterocyclic group, aryl group or heteroaryl group, wherein said alkyl group, cycloalkyl group, heterocyclic group, aryl group or heteroaryl group is optional is further selected by one or more of hydroxy, halogen, nitro, cyano, alkyl, alkoxy, haloalkyl, haloalkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl, =O , -C(O)R ⁸ , -C(O)OR ⁸ , -OC(O)R ⁸ , -NR ⁹ R ¹⁰ , -C(O)NR ⁹ R ¹⁰ , -SO ₂ NR ⁹ R ¹⁰ or - Substituted by the substituent of NR ⁹ C(O)R ¹⁰ ;

R ⁶ and R ⁷ are each independently selected from a hydrogen atom, a hydroxyl group, a halogen, an alkyl group, an alkoxy group, a cycloalkyl group, a heterocyclic group, an aryl group or a heteroaryl group, wherein the alkyl group, alkoxy group, A cycloalkyl, heterocyclyl, aryl or heteroaryl group is optionally further selected from one or more groups selected from hydroxy, halogen, nitro, cyano, alkyl, alkoxy, cycloalkyl, heterocyclyl, Aryl, heteroaryl, =O, -C(O)R ⁸ , -C(O)OR ⁸ , -OC(O)R ⁸ , -NR ⁹ R ¹⁰ , -C(O)NR ⁹ R ¹⁰ , -SO ₂ NR ⁹ R ¹⁰ or -NR ⁹ C(O)R ¹⁰ substituent;

Alternatively, R ⁶ and R ⁷ together with the atoms to which they are attached form a 4- to 8-membered heterocyclyl group containing one or more N, O or S(O) _r within the 4- to 8-membered heterocyclyl group, and The 4-8 membered heterocyclic group is optionally further selected from one or more groups selected from hydroxyl, halogen, nitro, cyano, alkyl, alkoxy, cycloalkyl, heterocyclic, aryl, heterocyclic Aryl, =O, -C(O)R ⁸ , -C(O)OR ⁸ , -OC(O)R ⁸ , -NR ⁹ R ¹⁰ , -C(O)NR ⁹ R ¹⁰ , -SO ₂ NR ⁹ R ¹⁰ or -NR ⁹ C(O) R ¹⁰ is substituted with a substituent;

wherein, R ⁸ , R ⁹ and R ¹⁰ are each independently selected from a hydrogen atom, an alkyl group, an amino group, a cycloalkyl group, a heterocyclic group, an aryl group or a heteroaryl group, wherein the alkyl group, cycloalkyl group, hetero The cyclyl, aryl or heteroaryl group is optionally further selected from one or more groups selected from hydroxy, halogen, nitro, amino, cyano, alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl, Substituted by substituents of heteroaryl, carboxyl or carboxylate;

r is 0, 1 or 2.

The compounds of the present invention may contain asymmetric centers or chiral centers and therefore exist in different stereoisomeric forms. All stereoisomeric forms of the compounds of the present invention are contemplated, including but not limited to diastereomers, enantiomers, and atropisomers and geometric (conformational) isomers and Their mixtures, such as racemic mixtures, are within the scope of the present invention.

Unless otherwise indicated, the structures depicted herein also include all isomeric (eg, diastereomeric, enantiomeric, and atropisomeric and geometric (conformational) isomeric forms of such structures; for example, , the R and S configuration of each asymmetric center, the (Z) and (E) double bond isomers, and the (Z) and (E) conformational isomers. Thus the individual stereoisomers of the compounds of the present invention as well as Enantiomeric mixtures, diastereomeric mixtures and geometric (conformational) isomer mixtures are all within the scope of the present invention.

"Pharmaceutically acceptable salts" refers to certain salts of the above-mentioned compounds that retain their original biological activity and are suitable for medicinal use. The pharmaceutically acceptable salt of the compound represented by the formula (I) may be a metal salt or an amine salt with a suitable acid.

"Pharmaceutical composition" means a mixture containing one or more of the compounds described herein, or a physiologically pharmaceutically acceptable salt or prodrug thereof, with other chemical components, and optionally other components such as physiologically pharmaceutically acceptable carriers and excipients. The purpose of the pharmaceutical composition is to facilitate the administration to the organism, facilitate the absorption of the active ingredient and then exert the biological activity.

Synthetic method of the compound of the present invention

In order to accomplish the purpose of the present invention, the present invention adopts following technical scheme:

The preparation method of the compound described in the general formula (II) of the present invention or its stereoisomer, tautomer or its pharmaceutically acceptable salt comprises the following steps:

The compound of general formula (IIA) is reacted with the compound of general formula (IIB) under basic conditions, and optionally the protective group is further removed to obtain the compound of general formula (II);

in:

X ¹ is a leaving group, and the leaving group is preferably chlorine, bromine, iodine, OMs or OTs; more preferably chlorine;

Ring B, R ¹ to R ³ , X, Y, G, E and n are as defined in the general formula (II).

detailed description

The following examples are used to further describe the present invention, but these examples do not limit the scope of the present invention.

Example

The examples give the preparation and related structural identification data of the representative compounds represented by formula (I). It must be noted that the following examples are used to illustrate the present invention rather than limit it. The ¹ H NMR spectrum was measured with a Bruker instrument (400 MHz), and the chemical shifts were expressed in ppm. A tetramethylsilane internal standard (0.00 ppm) was used. ¹ HNMR representation: s=singlet, d=doublet, t=triplet, m=multiplet, br=broadened, dd=doublet of doublet, dt=doublet of triplet. When coupling constants are provided, they are in Hz.

Mass spectrum is obtained by LC/MS instrument, and the ionization mode can be ESI or APCI.

The thin layer chromatography silica gel plate uses Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plate, the size of the silica gel plate used for thin layer chromatography (TLC) is 0.15mm ~ 0.2mm, and the size of the TLC separation and purification products is 0.4mm ~0.5mm.

Column chromatography generally uses Yantai Huanghai silica gel 200-300 mesh silica gel as the carrier.

In the following examples, all temperatures are in degrees Celsius unless otherwise indicated, and various starting materials and reagents were obtained from commercially available or synthesized according to known methods without further purification. For direct use, unless otherwise specified, commercially available manufacturers include but are not limited to Shanghai Haohong Biomedical Technology Co., Ltd., Shanghai Shaoyuan Reagent Co., Ltd., Shanghai Bide Pharmaceutical Technology Co., Ltd., Saen Chemical Technology (Shanghai) Co., Ltd. and Shanghai Lingkai Pharmaceutical Technology Co., Ltd., etc.

_CD3OD : Deuterated methanol.

CDCl ₃ : deuterated chloroform.

DMSO-d ₆ : deuterated dimethyl sulfoxide.

Unless otherwise specified in the examples, the solution in the reaction refers to an aqueous solution.

The compound was purified using column chromatography and thin layer chromatography eluent system, wherein the system was selected from: A: petroleum ether and ethyl acetate system; B: dichloromethane and methanol system; C: dichloromethane and ethyl acetate system; D: dichloromethane and ethanol system; E: ethyl acetate and tetrahydrofuran system; the volume ratio of the solvent varies according to the polarity of the compound, and a small amount of acidic or basic reagent can also be added to adjust , such as acetic acid or triethylamine, etc.

Room temperature: 20℃～30℃.

Example 1

1-(3-(8-Chloro-6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-1H-[1,2,3]triazolo[4,5-c]quinoline- 1-yl)azetidin-1-yl)prop-2-en-1-one

first step

3-((7-Bromo-6-chloro-8-fluoro-3-nitroquinolin-4-yl)amino)azetidine-1-carboxylate tert-butyl ester

7-Bromo-4,6-dichloro-8-fluoro-3-nitroquinoline 1a (550 mg, 1.62 mmol, prepared according to published patent WO2019110751) and tertiary 3-aminoazetidine-1-carboxylic acid Butyl ester 1b (417.98 mg, 2.43 mmol) was dissolved in acetonitrile (5 mL), cooled to 0 °C, N,N-diisopropylethylamine (627.32 mg, 4.85 mmol) was added dropwise, and the reaction was continued at room temperature for 6 Hour. Concentrated under reduced pressure, the obtained residue was separated and purified by silica gel column chromatography (eluent: A system) to obtain 3-((7-bromo-6-chloro-8-fluoro-3-nitroquinoline-4 -yl)amino)azetidine-1-carboxylate tert-butyl ester 1c (400 mg, 840.87 μmol), yield: 51.91%.

MS m/z(ESI): 474.7[M+1] ⁺

second step

3-((3-Amino-7-bromo-6-chloro-8-fluoroquinolin-4-yl)amino)azetidine-1-carboxylate tert-butyl ester

3-((7-Bromo-6-chloro-8-fluoro-3-nitroquinolin-4-yl)amino)azetidine-1-carboxylate tert-butyl ester 1c (1.2 g, 2.52 mmol ), ammonium chloride (674.67 mg, 12.61 mmol) and iron powder (704.44 mg, 12.61 mmol) were dissolved in a mixed solvent of methanol (10 mL) and water (2 mL), heated to 90° C. to react for 5 hours. After the reaction was completed, it was filtered while hot, and the filtrate was concentrated under reduced pressure to remove methanol. The system was extracted with ethyl acetate (100 mL×2), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain crude 3-((3-amino-7-bromo-6-chloro-8-fluoro. Quinolin-4-yl)amino)azetidine-1-carboxylate tert-butyl ester 1d (950 mg, 2.13 mmol), yield: 84.49%.

MS m/z(ESI): 445.0[M+1] ⁺

third step

tert-butyl

3-(7-Bromo-8-chloro-6-fluoro-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)azetidine-1-carboxy tert-butyl acid

3-((3-Amino-7-bromo-6-chloro-8-fluoroquinolin-4-yl)amino)azetidine-1-carboxylate tert-butyl ester 1d (950 mg, 2.13 mmol) was dissolved in In a mixed solvent of acetic acid (10 mL) and water (2 mL), cooled to 0 °C, sodium nitrite (220.60 mg, 3.20 mmol) was added, the reaction was continued at 0 °C for 0.5 hours, and the temperature was raised to room temperature for 2 hours. After the reaction, the system was adjusted to basicity with saturated sodium carbonate solution, extracted with ethyl acetate (100 mL×2), the organic phases were combined, washed with saturated brine (100 mL×3), dried over anhydrous sodium sulfate, filtered, Concentrated under reduced pressure, the obtained residue was separated and purified by silica gel column chromatography (eluent: system A) to obtain 3-(7-bromo-8-chloro-6-fluoro-1H-[1,2,3] Triazolo[4,5-c]quinolin-1-yl)azetidine-1-carboxylate tert-butyl ester 1e (520 mg, 1.14 mmol), yield: 53.42%.

MS m/z(ESI): 455.8[M+1] ⁺

the fourth step

3-(8-Chloro-6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl ) tert-butyl azetidine-1-carboxylate

3-(7-Bromo-8-chloro-6-fluoro-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)azetidine-1- tert-Butyl carboxylate 1e (350 mg, 766.37 μmol), (2-fluoro-6-hydroxyphenyl)boronic acid 1f (262.88 mg, 1.69 mmol), potassium phosphate (325.34 mg, 1.53 mmol) and X-PHOS Pd G2 ( 120.44 mg, 153.27 μmol) was dissolved in tetrahydrofuran (10 mL) under argon protection, heated to 50° C., and reacted overnight. After the reaction is completed, filter and concentrate the filtrate under reduced pressure, and the obtained residue is separated and purified by silica gel column chromatography (eluent: A system) to obtain 3-(8-chloro-6-fluoro-7-(2- Fluoro-6-hydroxyphenyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)azetidine-1-carboxylate tert-butyl ester 1g ( 20 mg, 40.99 μmol), yield: 5.35%.

MS m/z(ESI): 487.9[M+1] ⁺

the fifth step

2-(1-(azetidin-3-yl)-8-chloro-6-fluoro-1H-[1,2,3]triazolo[4,5-c]quinolin-7-yl )-3-Fluorophenol

3-(8-Chloro-6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-1H-[1,2,3]triazolo[4,5-c]quinoline-1- 1 g (15 mg, 30.74 μmol) of tert-butyl azetidine-1-carboxylate was dissolved in dichloromethane (5 mL), and a solution of HCl in 1,4-dioxane (4 M, 3 mL) was added, The reaction was carried out at room temperature for 2 hours. After the reaction, concentrated under reduced pressure to obtain crude 2-(1-(azetidin-3-yl)-8-chloro-6-fluoro-1H-[1,2,3]triazolo[4, 5-c]Quinolin-7-yl)-3-fluorophenol 1 h (11 mg, 28.37 μmol), yield: 92.27%.

MS m/z(ESI): 388.0[M+1] ⁺

Step 6

2-(1-(1-Acryloylazetidin-3-yl)-8-chloro-6-fluoro-1H-[1,2,3]triazolo[4,5-c]quinoline -7-yl)-3-fluorophenylacrylate

2-(1-(azetidin-3-yl)-8-chloro-6-fluoro-1H-[1,2,3]triazolo[4,5-c]quinoline-7- base)-3-fluorophenol for 1h (15.9mg, 41.00μmol) was dissolved in dichloromethane (5mL) for 1h, triethylamine (16.60mg, 164.01μmol) was added, cooled to 0°C, and acryloyl chloride (7.42mg) was added dropwise , 82.01 μmol) in dichloromethane (2 mL), and the reaction was continued for 1 hour at 0 °C. The reaction solution was concentrated under reduced pressure to obtain crude 2-(1-(1-acryloylazetidin-3-yl)-8-chloro-6-fluoro-1H-[1,2,3]triazolo [4,5-c]quinolin-7-yl)-3-fluorophenylacrylate 1i (20.3 mg, 40.99 μmol) was used directly in the next reaction.

MS m/z(ESI): 496.0[M+1] ⁺

Step 7

1-(3-(8-Chloro-6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-1H-[1,2,3]triazolo[4,5-c]quinoline- 1-yl)azetidin-1-yl)prop-2-en-1-one

2-(1-(1-Acryloylazetidin-3-yl)-8-chloro-6-fluoro-1H-[1,2,3]triazolo[4,5-c]quinoline Lin-7-yl)-3-fluorophenylacrylate 1i (20 mg, 40.33 μmol) was dissolved in a mixed solvent of tetrahydrofuran (5 mL) and water (2 mL), and lithium hydroxide monohydrate (8.46 mg, 201.67 μmol) was added. ), reacted under ice bath for 0.5 h. Concentrate under reduced pressure to remove tetrahydrofuran, extract with dichloromethane (60 mL×2), combine the organic phases, dry over anhydrous sodium sulfate, filter, and concentrate under reduced pressure. The obtained residue is separated and purified by preparative HPLC (separation column: Boston Prime C18, 150 x 30 mm ID, 5 μm; mobile phase A: water (0.05% _{NH3H2O +} 10 mM _{NH4HCO3 )} , mobile phase B: acetonitrile; flow rate: 25 mL/min) to give 1-(3-( 8-Chloro-6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)aza Cyclobutan-1-yl)prop-2-en-1-one 1 (6 mg, 12.22 μmol), yield: 30.30%.

MS m/z(ESI): 442.0[M+1] ⁺

Example 2

1-(3-(8-Chloro-6-fluoro-7-(3-hydroxynaphthalen-1-yl)-1H-[1,2,3]triazolo[4,5-c]quinoline-1 -yl)azetidin-1-yl)prop-2-en-1-one

first step

3-(8-Chloro-6-fluoro-7-(3-hydroxynaphthalen-1-yl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl) tert-butyl azetidine-1-carboxylate

3-(7-Bromo-8-chloro-6-fluoro-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)azetidine-1- tert-Butyl carboxylate 1e (354.89 mg, 1.31 mmol), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2-yl)naphthalene-2 -Alcohol 2a (300 mg, 656.89 μmol), potassium phosphate (418.30 mg, 1.97 mmol) and X-PHOS Pd G2 (51.62 mg, 65.69 μmol) were dissolved in tetrahydrofuran (5 mL), protected by argon, heated to 50°C, and reacted overnight. After the reaction was completed, concentrated under reduced pressure, and the obtained residue was separated and purified by silica gel column chromatography (eluent: A system) to obtain 3-(8-chloro-6-fluoro-7-(3-hydroxynaphthalene-1). -yl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)azetidine-1-carboxylate tert-butyl ester 2b (340 mg, 653.90 μmol) , Yield: 99.55%.

MS m/z(ESI): 520.2[M+1] ⁺

second step

4-(1-(azetidin-3-yl)-8-chloro-6-fluoro-1H-[1,2,3]triazolo[4,5-c]quinolin-7-yl ) Naphthalene-2-ol

3-(8-Chloro-6-fluoro-7-(3-hydroxynaphthalen-1-yl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl ) tert-butyl azetidine-1-carboxylate 2b (100 mg, 192.32 μmol) was dissolved in dichloromethane (4 mL), a solution of HCl in 1,4-dioxane (4 M, 3 mL) was added, room temperature React for 1 hour. After the reaction, concentrated under reduced pressure to obtain crude product 4-(1-(azetidin-3-yl)-8-chloro-6-fluoro-1H-[1,2,3]triazolo[4, 5-c]Quinolin-7-yl)naphthalen-2-ol 2c (80 mg, 190.55 μmol), yield: 99.08%.

MS m/z(ESI): 420.1[M+1] ⁺

third step

1-(3-(8-Chloro-6-fluoro-7-(3-hydroxynaphthalen-1-yl)-1H-[1,2,3]triazolo[4,5-c]quinoline-1 -yl)azetidin-1-yl)prop-2-en-1-one

4-(1-(azetidin-3-yl)-8-chloro-6-fluoro-1H-[1,2,3]triazolo[4,5-c]quinoline-7- base)naphthalene-2-ol 2c (80mg, 190.55μmol) was dissolved in dichloromethane (5mL), cooled to 0°C, triethylamine (77.13mg, 762.20μmol) was added, acryloyl chloride (17.25mg, 190.55μmol) was added dropwise μmol) in dichloromethane (2 mL), the reaction was continued for 1 hour at 0°C. After the reaction, the system was washed with saturated ammonium chloride solution (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The obtained residue was separated and purified by preparative HPLC (separation column: Boston Prime C18, 150×30 mm ID). , 5 μm; mobile phase A: water (0.05% NH ₃ H ₂ O + 10 mM NH ₄ HCO ₃ ), mobile phase B: acetonitrile; flow rate: 25 mL/min) to give 1-(3-(8-chloro-6- Fluoro-7-(3-hydroxynaphthalen-1-yl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)azetidin-1-yl ) prop-2-en-1-one 2 (25 mg, 48.93 μmol), yield: 25.68%.

MS m/z(ESI): 474.1[M+1] ⁺

Example 3

1-(3-(7-(2-Amino-7-fluorobenzo[d]thiazol-4-yl)-8-chloro-6-fluoro-1H-[1,2,3]triazolo[4 ,5-c]quinolin-1-yl)azetidine-1-yl)prop-2-en-1-one

first step

(7-Fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2-yl)benzo[d]thiazol-2-yl)amino tert-butyl formate

2-((tert-butoxycarbonyl)amino)-7-fluorobenzo[d]thiazol-4-yl trifluoromethanesulfonate 3a (8.5 g, 20.41 mmol, prepared according to published patent US 20200115375), acetic acid Potassium (6.01 g, 61.24 mmol), pinacol diboronate (41.47 g, 163.32 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium dichloromethane complex (2.81 g, 3.27 mmol) was dissolved in 1,4-dioxane (200 mL) and reacted at 85° C. for 16 hours under nitrogen protection. The progress of the reaction was monitored by LC-MS. After the reaction, 300 mL of water was added to dilute the reaction solution, extracted with ethyl acetate (300 mL×2), the organic phases were combined, washed with saturated brine (300 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the obtained The residue was separated and purified by silica gel column chromatography (eluent: E system) to obtain (7-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxane) Pentaboran-2-yl)benzo[d]thiazol-2-yl)carbamate tert-butyl ester 3b (1.7 g), yield: 21.12%.

MS m/z(ESI): 256.9[M+1-139] ⁺

¹ H NMR (400MHz, DMSO-d ₆ ) δ 1.32(s, 12H), 1.51(s, 9H), 7.15(t, J=8.66Hz, 1H), 7.76(t, J=7.28Hz, 1H) ,12.14(br s,1H).

second step

3-(7-(2-((tert-butoxycarbonyl)amino)-7-fluorobenzo[d]thiazol-4-yl)-8-chloro-6-fluoro-1H-[1,2,3 ]Triazolo[4,5-c]quinolin-1-yl)azetidine-1-carboxylate tert-butyl ester

3-(7-Bromo-8-chloro-6-fluoro-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)azetidine-1- Carboxylic acid tert-butyl ester 1e (512 mg, 1.30 mmol), (7-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2-yl) ) benzo[d]thiazol-2-yl)carbamate tert-butyl ester 3b (539.15 mg, 1.18 mmol), potassium phosphate (501.17 mg, 2.36 mmol), X-PHOS Pd G2 (185.53 mg, 236.11 μmol) were dissolved in In tetrahydrofuran (10 mL), under argon protection, heated to 50°C, and reacted for 3 hours. After the reaction was completed, it was concentrated under reduced pressure, and the obtained residue was separated and purified by silica gel column chromatography (eluent: A system) to obtain 3-(7-(2-((tert-butoxycarbonyl)amino)-7 -Fluorobenzo[d]thiazol-4-yl)-8-chloro-6-fluoro-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)azepine Cyclobutane-1-carboxylate tert-butyl ester 3c (270 mg, 419.20 μmol), yield: 32.23%.

MS m/z(ESI): 443.8[M+1] ⁺

third step

4-(1-(azetidin-3-yl)-8-chloro-6-fluoro-1H-[1,2,3]triazolo[4,5-c]quinolin-7-yl )-7-Fluorobenzo[d]thiazol-2-amine

3-(7-(2-((tert-butoxycarbonyl)amino)-7-fluorobenzo[d]thiazol-4-yl)-8-chloro-6-fluoro-1H-[1,2, 3] Triazolo[4,5-c]quinolin-1-yl)azetidine-1-carboxylate tert-butyl ester 3c (270 mg, 419.20 μmol) was dissolved in dichloromethane (8 mL) and added A solution of HCl in 1,4-dioxane (4M, 8 mL) was reacted at room temperature for 1 hour. After the reaction, concentrated under reduced pressure to obtain crude product 4-(1-(azetidin-3-yl)-8-chloro-6-fluoro-1H-[1,2,3]triazolo[4, 5-c]quinolin-7-yl)-7-fluorobenzo[d]thiazol-2-amine 3d (186 mg, 419.05 μmol) was used directly in the next reaction.

MS m/z(ESI): 443.8[M+1] ⁺

the fourth step

1-(3-(7-(2-Amino-7-fluorobenzo[d]thiazol-4-yl)-8-chloro-6-fluoro-1H-[1,2,3]triazolo[4 ,5-c]quinolin-1-yl)azetidine-1-yl)prop-2-en-1-one

4-(1-(azetidin-3-yl)-8-chloro-6-fluoro-1H-[1,2,3]triazolo[4,5-c]quinoline-7- base)-7-fluorobenzo[d]thiazol-2-amine 3d (186 mg, 419.05 μmol) was dissolved in dichloromethane (3 mL), added triethylamine (127.21 mg, 1.26 mmol), cooled to 0 °C, A solution of acryloyl chloride (37.93 mg, 419.05 μmol) in dichloromethane (2 mL) was added dropwise, and the reaction was continued at 0° C. for 1 hour. After the reaction, the system was washed with saturated ammonium chloride solution (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The obtained residue was separated and purified by preparative HPLC (separation column: Boston Prime C18, 150×30 mm ID). , 5 μm; mobile phase A: water (0.05% NH ₃ H ₂ O + 10 mM NH ₄ HCO ₃ ), mobile phase B: acetonitrile; flow rate: 25 mL/min) to give 1-(3-(7-(2-amino) -7-Fluorobenzo[d]thiazol-4-yl)-8-chloro-6-fluoro-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl) Azetidine-1-yl)prop-2-en-1-one 3 (100 mg, 200.84 μmol), yield: 47.93%.

MS m/z(ESI): 498.0[M+1] ⁺

Examples 3A and 3B

1-(3-(7-(2-Amino-7-fluorobenzo[d]thiazol-4-yl)-8-chloro-6-fluoro-1H-[1,2,3]triazolo[ 4,5-c]Quinolin-1-yl)azetidin-1-yl)prop-2-en-1-one 3 (250 mg, 501.9 μmol) was chiral resolved by SFC (column model: ChiralPak AD, 250×30mm ID, 10μm; mobile phase: A for CO ₂ and B for Isopropanol (0.1%NH3H2O); column pressure: 100bar; flow rate: 70mL/min; detection wavelength: 220nm; column temperature: 38℃) after purification , to obtain compound 3A and compound 3B, which belong to one of the single configuration compounds (shorter retention time) and single configuration compounds (longer retention time), respectively.

Single configuration compound (shorter retention time):

MS m/z(ESI): 498.1[M+1] ⁺

80 mg; retention time 2.792 minutes, chiral purity 100% ee.

Single configuration compound (longer retention time):

136mg; retention time 3.908 minutes, chiral purity 99.08%ee.

MS m/z(ESI): 498.2[M+1] ⁺

Example 4

1-(3-(7-(2-Amino-5,7-difluorobenzo[d]thiazol-4-yl)-8-chloro-6-fluoro-1H-pyrazolo[4,3-c ]quinolin-1-yl)azetidin-1-yl)prop-2-en-1-one

first step

(7-bromo-4,6-dichloro-8-fluoroquinolin-3-yl)methanol

(7-Bromo-4,6-dichloro-8-fluoroquinolin-3-yl)methanol

7-Bromo-4,6-dichloro-8-fluoroquinoline-3-carboxylic acid ethyl ester 4a (0.05g, 136.24μmol, prepared according to published patent WO2016164675A1) was dissolved in tetrahydrofuran (2mL), cooled to -78 °C, lithium diisopropylamide (1M, 299.73 μL) was added dropwise, the temperature was slowly raised to 25 °C, and the reaction was continued for 3 hours. The reaction was complete as detected by LC-MS. 5 mL of water was added to the reaction solution to quench the reaction, extracted with ethyl acetate (5 mL × 3), the organic phases were combined, dried over anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure to obtain a yellow oily crude product (7-bromo-4, 6-Dichloro-8-fluoroquinolin-3-yl)methanol 4b (20 mg, 61.55 μmol), 45.17% yield.

MS m/z(ESI): 225.9[M+3] ⁺

second step

7-Bromo-4,6-dichloro-8-fluoroquinoline-3-carbaldehyde

(7-Bromo-4,6-dichloro-8-fluoroquinolin-3-yl)methanol 4b (0.025 g, 76.93 μmol) was dissolved in dichloromethane (2 mL), and manganese dioxide ( 66.88 mg, 769.32 μmol), heated to 40 °C for 16 hours. The reaction was complete as detected by LC-MS. Filtration and concentration under reduced pressure gave crude 7-bromo-4,6-dichloro-8-fluoroquinoline-3-carbaldehyde 4c.

MS m/z(ESI): 223.6[M+3] ⁺

third step

tert-butyl

3-(7-Bromo-8-chloro-6-fluoro-1H-pyrazolo[4,3-c]quinolin-1-yl)azetidine-1-carboxylate tert-butyl ester

7-Bromo-4,6-dichloro-8-fluoroquinoline-3-carbaldehyde 4c (500 mg, 1.55 mmol) and tert-butyl 3-hydrazinoazetidine-1-carboxylate 4d (434.84 mg , 2.32mmol, prepared according to published patent WO2012004743A1) was dissolved in ethanol (6mL), added triethylamine (470.00mg, 4.64mmol), and reacted at 80°C for 3 hours. The reaction was complete as detected by LC-MS. Concentrated under reduced pressure, the obtained residue was separated and purified by silica gel column chromatography (eluent: system A) to obtain 3-(7-bromo-8-chloro-6-fluoro-1H-pyrazolo[4,3- c] Quinolin-1-yl)azetidine-1-carboxylate tert-butyl ester 4e (0.2 g, 438.88 μmol), 28.35% yield.

MS m/z(ESI): 455.0[M+1] ⁺

the fourth step

3-(7-(2-((tert-butoxycarbonyl)amino)-5,7-difluorobenzo[d]thiazol-4-yl)-8-chloro-6-fluoro-1H-pyrazolo [4,3-c]Quinolin-1-yl)azetidine-1-carboxylate tert-butyl ester

3-(7-Bromo-8-chloro-6-fluoro-1H-pyrazolo[4,3-c]quinolin-1-yl)azetidine-1-carboxylate tert-butyl ester 4e ( 300mg, 658.32μmol), (2-((tert-butoxycarbonyl)amino)-5,7-difluorobenzo[d]thiazol-4-yl)boronic acid 4f (260.78mg, 789.98μmol, according to published patent US20200115375A1 prepared), potassium phosphate (279.47 mg, 1.32 mmol) and X-PHOS Pd G2 (51.73 mg, 65.83 μmol) were dissolved in tetrahydrofuran (8 mL) and reacted at 50°C overnight. Cooled to room temperature, the reaction solution was filtered with celite, the filtrate was concentrated under reduced pressure, an appropriate amount of methanol was added to dissolve the residue, filtered again, the filtrate was concentrated under reduced pressure, and the obtained residue was separated and purified by silica gel column chromatography (eluent). : A system) to give 3-(7-(2-((tert-butoxycarbonyl)amino)-5,7-difluorobenzo[d]thiazol-4-yl)-8-chloro-6-fluoro -1H-pyrazolo[4,3-c]quinolin-1-yl)azetidine-1-carboxylate tert-butyl ester 4 g (21 mg, 31.77 μmol), 4.83% yield.

MS m/z(ESI): 661.2[M+1] ⁺

the fifth step

4-(1-(azetidin-3-yl)-8-chloro-6-fluoro-1H-pyrazolo[4,3-c]quinolin-7-yl)-5,7-di Fluorobenzo[d]thiazol-2-amine

3-(7-(2-((tert-butoxycarbonyl)amino)-5,7-difluorobenzo[d]thiazol-4-yl)-8-chloro-6-fluoro-1H-pyrazole [4,3-c]quinolin-1-yl)azetidine-1-carboxylate tert-butyl ester 4g (21mg, 31.77μmol) was dissolved in dichloromethane (3mL), HCl/1 was added, 4-Dioxane solution (4M, 3 mL) was reacted at room temperature for 1 hour. Concentration under reduced pressure gave crude 4-(1-(azetidin-3-yl)-8-chloro-6-fluoro-1H-pyrazolo[4,3-c]quinolin-7-yl) -5,7-Difluorobenzo[d]thiazol-2-amine 4h (14.6 mg, 25.34 μmol), 79.78% yield.

MS m/z(ESI): 463.0[M+3] ⁺

Step 6

1-(3-(7-(2-Amino-5,7-difluorobenzo[d]thiazol-4-yl)-8-chloro-6-fluoro-1H-pyrazolo[4,3-c ]quinolin-1-yl)azetidin-1-yl)prop-2-en-1-one

4-(1-(azetidin-3-yl)-8-chloro-6-fluoro-1H-pyrazolo[4,3-c]quinolin-7-yl)-5,7- Difluorobenzo[d]thiazol-2-amine 4h (14.6 mg, 25.34 μmol) was dissolved in dichloromethane (5 mL), cooled to 0°C, triethylamine (8.56 mg, 84.62 μmol) and acryloyl chloride ( 2.55 mg, 28.21 μmol), and the reaction was continued for 1 hour at 0 °C. After the reaction, the reaction solution was washed with saturated ammonium chloride solution (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The obtained residue was separated and purified by preparative HPLC (separation column: Boston Prime C18, 150 × 30 mm). ID, 5 μm; mobile phase A: water (0.05% _{NH3H2O +} 10 mM _{NH4HCO3 )} , mobile phase B: acetonitrile; flow rate: 25 mL/min) to give 1-(3-(7-(2- Amino-5,7-difluorobenzo[d]thiazol-4-yl)-8-chloro-6-fluoro-1H-pyrazolo[4,3-c]quinolin-1-yl)azacycle Butan-1-yl)prop-2-en-1-one 4 (1 mg, 1.84 μmol), 6.54% yield.

MS m/z(ESI): 516.0[M+2] ⁺

Example 5

1-(3-(7-(2-Amino-5,7-difluorobenzo[d]thiazol-4-yl)-8-chloro-6-fluoro-1H-[1,2,3]triazole [4,5-c]quinolin-1-yl)azetidin-1-yl)prop-2-en-1-one

first step

3-(7-(2-((tert-butoxycarbonyl)amino)-5,7-difluorobenzo[d]thiazol-4-yl)-8-chloro-6-fluoro-1H-[1,2 ,3]Triazolo[4,5-c]quinolin-1-yl)azetidine-1-carboxylate tert-butyl ester

(2-((tert-butoxycarbonyl)amino)-5,7-difluorobenzo[d]thiazol-4-yl)boronic acid 4f (426 mg, 1.29 mmol), 3-(7-bromo-8- Chloro-6-fluoro-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)azetidine-1-carboxylate tert-butyl ester 1e (491.12 mg, 1.08 mmol), potassium phosphate (456.52 mg, 2.15 mmol) and X-PHOS Pd G2 (84.50 mg, 107.54 μmol) were dissolved in tetrahydrofuran (6 mL), under argon protection, heated to 50 °C and reacted overnight. After the reaction, the filtrate was concentrated under reduced pressure, and the obtained residue was separated and purified by preparative HPLC (separation column: AKZONOBEL Kromasil; 250×21.2 mm ID; 5 μm; mobile phase A: 0.05% TFA+H2O, mobile phase B: Acetonitrile; flow rate: 20 mL/min) to give the product 3-(7-(2-((tert-butoxycarbonyl)amino)-5,7-difluorobenzo[d]thiazol-4-yl)-8-chloro -6-Fluoro-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)azetidine-1-carboxylate tert-butyl ester 5a (100 mg, 151.04 μmol ) in 14.05% yield.

MS m/z(ESI): 661.8[M+1] ⁺

second step

4-(1-(azetidin-3-yl)-8-chloro-6-fluoro-1H-[1,2,3]triazolo[4,5-c]quinolin-7-yl )-5,7-difluorobenzo[d]thiazol-2-amine

3-(7-(2-((tert-butoxycarbonyl)amino)-5,7-difluorobenzo[d]thiazol-4-yl)-8-chloro-6-fluoro-1H-[1, 2,3]Triazolo[4,5-c]quinolin-1-yl)azetidine-1-carboxylate tert-butyl ester 5a (143 mg, 215.99 μmol) was dissolved in dichloromethane (5 mL) , 1,4-dioxane solution of hydrochloric acid (4M, 3 mL) was added, and the reaction was carried out at room temperature for 1 hour. Concentration under reduced pressure gave crude product 4-(1-(azetidin-3-yl)-8-chloro-6-fluoro-1H-[1,2,3]triazolo[4,5-c ]quinolin-7-yl)-5,7-difluorobenzo[d]thiazol-2-amine 5b (99.75 mg, 215.98 μmol), yield: 100%.

MS m/z(ESI): 461.8[M+1] ⁺

third step

1-(3-(7-(2-Amino-5,7-difluorobenzo[d]thiazol-4-yl)-8-chloro-6-fluoro-1H-[1,2,3]triazole [4,5-c]quinolin-1-yl)azetidin-1-yl)prop-2-en-1-one

4-(1-(azetidin-3-yl)-8-chloro-6-fluoro-1H-[1,2,3]triazolo[4,5-c]quinoline-7- base)-5,7-difluorobenzo[d]thiazol-2-amine 5b (99.75 mg, 215.98 μmol) was dissolved in dichloromethane (3 mL), cooled to 0 °C, added triethylamine (65.56 mg, 647.94 μmol) and acryloyl chloride (19.55 mg, 215.98 μmol), the reaction was continued for 1 hour at 0°C. After the reaction, the reaction solution was concentrated under reduced pressure, and the obtained residue was separated and purified by preparative HPLC (separation column: AKZONOBEL Kromasil; 250×21.2 mm ID; 5 μm; mobile phase A: 0.05% TFA+H2O, mobile phase B: acetonitrile; Flow rate: 20 mL/min) to give the product 1-(3-(7-(2-amino-5,7-difluorobenzo[d]thiazol-4-yl)-8-chloro-6-fluoro-1H- [1,2,3]Triazolo[4,5-c]quinolin-1-yl)azetidin-1-yl)prop-2-en-1-one 5 (30 mg, 58.15 μmol) , Yield: 26.92%.

MS m/z(ESI): 516.1[M+1] ⁺

¹ H NMR (400MHz, Chloroform-d) δ9.62(s,1H), 8.08(s,1H), 7.96-7.46(m,2H), 6.95(s,1H), 6.56-6.22(m,2H) ,6.04(t,J＝6.7Hz,1H),5.82(d,J＝10.3Hz,1H),5.43–4.64(m,4H).

Examples 5A and 5B

1-(3-(7-(2-Amino-5,7-difluorobenzo[d]thiazol-4-yl)-8-chloro-6-fluoro-1H-[1,2,3]tris Azolo[4,5-c]quinolin-1-yl)azetidin-1-yl)prop-2-en-1-one 5 (30 mg, 58.15 μmol) was chiral resolved by SFC (column Model: ChiralCel OD, 250×30mm ID, 10μm; mobile phase: A for CO2 and B for Ethanol; column pressure: 100bar; flow rate: 80mL/min; detection wavelength: 220nm; Compound 5A and Compound 5B belong to one of the single configuration compound (shorter retention time) and the single configuration compound (longer retention time).

Single configuration compound (shorter retention time):

MS m/z(ESI): 516.1[M+1] ⁺

13 mg; retention time 5.026 minutes, chiral purity 100% ee.

Single configuration compound (longer retention time):

10mg; retention time 8.542 minutes, chiral purity 100%ee.

MS m/z(ESI): 516.1[M+1] ⁺

Example 6

1-(3-(7-(2-Amino-5-fluorobenzo[d]thiazol-4-yl)-8-chloro-6-fluoro-3-methyl-1H-pyrazolo[4,3 -c]quinolin-1-yl)azetidine-1-yl)prop-2-en-1-one

first step

1-(3-Fluoro-2-methoxyphenyl)thiourea

3-Fluoro-2-methoxyaniline 6a (20 g, 141.70 mmol) was added to tetrahydrofuran (500 mL), and a solution of benzoyl isothiocyanate 6b (23.13 g, 141.70 mmol) in tetrahydrofuran (100 mL) was slowly added dropwise. After the dropwise addition was completed, the reaction was continued at room temperature for 3 hours. The LCMS monitoring raw materials were all converted into intermediates. Water (80 mL) and sodium hydroxide (6.80 g, 170.04 mmol) were added, and the reaction was heated to 80 °C overnight. Cooled to room temperature, extracted with ethyl acetate (100 mL×1), the organic phase was washed with saturated brine (100 mL×1), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The obtained residue was subjected to silica gel column chromatography (washed). Removal agent: A system) was separated and purified to obtain the product 1-(3-fluoro-2-methoxyphenyl)thiourea 6c (22 g, 109.87 mmol), yield: 77.55%.

MS m/z(ESI): 201.1[M+1] ⁺

second step

5-Fluoro-4-methoxybenzo[d]thiazol-2-amine

1-(3-Fluoro-2-methoxyphenyl)thiourea 6c (7.09 g, 35.41 mmol) was added to acetic acid (200 mL), lithium bromide (4.61 g, 53.11 mmol) was added, and liquid bromine (5.77 mmol) was slowly added dropwise. g, 36.12 mmol), keeping the temperature below 30°C. After the dropwise addition, the temperature of the reaction solution was raised to 40°C for overnight reaction. The reaction solution was cooled, poured into water (500 mL), made alkaline with saturated sodium carbonate solution, extracted with ethyl acetate (300 mL×1), the organic phase was washed with saturated brine (100 mL×1), dried over anhydrous sodium sulfate, Filtration and concentration under reduced pressure, the obtained residue was separated and purified by silica gel column chromatography (eluent: A system) to obtain the product 5-fluoro-4-methoxybenzo[d]thiazol-2-amine 6d ( 7 g, 35.31 mmol), yield: 99.73%.

MS m/z(ESI): 199.1[M+1] ⁺

third step

2-Amino-5-fluorobenzo[d]thiazol-4-ol

5-Fluoro-4-methoxybenzo[d]thiazol-2-amine 6d (7 g, 35.31 mmol) was added to dichloromethane (70 mL), cooled to 0 °C, and boron tribromide (22.12 g) was added dropwise. , 88.29mmol, 8.51mL), after the dropwise addition was completed, the reaction was transferred to room temperature overnight. The reaction solution was poured into ice water (300 mL), made alkaline with saturated sodium carbonate solution, extracted with ethyl acetate (500 mL×1), the organic phase was washed with saturated brine (100 mL×1), dried over anhydrous sodium sulfate, filtered , concentrated under reduced pressure to obtain crude product 2-amino-5-fluorobenzo[d]thiazol-4-ol 6e (5.2 g, 28.23 mmol), yield: 79.94%.

MS m/z(ESI): 185.1[M+1] ⁺

the fourth step

tert-Butyl (4-((tert-butoxycarbonyl)oxy)-5-fluorobenzo[d]thiazol-2-yl)carbamate

2-Amino-5-fluorobenzo[d]thiazol-4-ol 6e (10 g, 54.29 mmol), dimethylaminopyridine (1.33 g, 10.86 mmol), triethylamine (10.99 g, 108.58 mmol, 15.13 mL) and di-tert-butyl dicarbonate (23.70 g, 108.58 mmol) were added to dichloromethane (80 mL) and reacted at room temperature for 4 hours. The reaction solution was concentrated under reduced pressure, ethyl acetate (100 mL) and water (50 mL) were added, the layers were separated, the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product (4-((tert-butoxycarbonyl) Oxy)-5-fluorobenzo[d]thiazol-2-yl)carbamate tert-butyl ester 6f (20 g, 52.03 mmol), yield: 95.83%.

MS m/z(ESI): 385.1[M+1] ⁺

the fifth step

tert-Butyl (5-fluoro-4-hydroxybenzo[d]thiazol-2-yl)carbamate

tert-Butyl (4-((tert-butoxycarbonyl)oxy)-5-fluorobenzo[d]thiazol-2-yl)carbamate 6f (20 g, 52.03 mmol) was added to tetrahydrofuran (80 mL) and water (20 mL) ), cooled to 0°C, added lithium hydroxide monohydrate (10.92 g, 260.13 mmol), and transferred to room temperature to react overnight. The reaction solution was added with ethyl acetate (100 mL) and water (50 mL), extracted with ethyl acetate (100 mL×2), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product (5-fluoro-4 - tert-butyl hydroxybenzo[d]thiazol-2-yl)carbamate 6 g (14.45 g, 50.83 mmol), yield: 97.69%.

MS m/z(ESI): 228.9[M+1-56] ⁺

Step 6

2-((tert-Butoxycarbonyl)amino)-5-fluorobenzo[d]thiazol-4-yl trifluoromethanesulfonate

Under ice bath, tert-butyl (5-fluoro-4-hydroxybenzo[d]thiazol-2-yl)carbamate 6 g (20 g, 70.35 mmol) was dissolved in dichloromethane (80 mL), followed by the addition of pyridine ( 11.13 g, 140.69 mmol, 11.36 mL), trifluoromethanesulfonic anhydride (23.82 g, 84.42 mmol, 14.26 mL), stirred for 30 minutes. The reaction solution was added with water (150 mL), extracted with dichloromethane (150 mL×3), and the organic phases were combined, washed with an aqueous solution of citric acid monohydrate (50 mL) and saturated brine (50 mL) in turn, dried over anhydrous sodium sulfate, filtered, and reduced The resulting residue was separated and purified by silica gel column chromatography (eluent: system A) to obtain the product 2-((tert-butoxycarbonyl)amino)-5-fluorobenzo[d]thiazole-4- triflate 6h (13.6 g, 32.66 mmol), yield: 46.43%.

MS m/z(ESI): 361.0[M+1-56] ⁺

Step 7

(2-((tert-butoxycarbonyl)amino)-5-fluorobenzo[d]thiazol-4-yl)boronic acid

2-((tert-butoxycarbonyl)amino)-5-fluorobenzo[d]thiazol-4-yl trifluoromethanesulfonate 6h (5 g, 12.01 mmol), pinacol biboronate (24.40 g, 96.07 mmol) was mixed with 1,4-dioxane (80 mL), potassium acetate (3.54 g, 36.03 mmol) was added, replaced with argon, stirred for 10 minutes, and [1,1'-bis(diphenylphosphine) was added base)ferrocene]palladium dichloride (2.64 g, 3.60 mmol), under argon protection, and stirred at 100° C. for 8 hours. Add pinacol biboronate (24.40g, 96.07mmol), [1,1'-bis(diphenylphosphino)ferrocene]dichloride palladium (2.64g, 3.60mmol), under argon protection, Stirring was continued at 100°C for 8 hours. The reaction solution was concentrated under reduced pressure, and the obtained residue was separated and purified by silica gel column chromatography (eluent: system A) to obtain the product (2-((tert-butoxycarbonyl)amino)-5-fluorobenzo[d] Thiazol-4-yl)boronic acid 6i (2 g, 6.41 mmol), yield: 53.36%.

MS m/z(ESI): 312.9[M+1] ⁺

Step 8

2-(((3-Bromo-4-chloro-2-fluorophenyl)amino)methylene)-3-oxobutyric acid ethyl ester

3-Bromo-4-chloro-2-fluoroaniline 6j (3g, 13.37mmol, prepared according to published patent WO2016164675A1), 2-(ethoxymethylene)-3-oxobutyric acid ethyl ester 6k (3.73g , 20.05 mmol) and mixed, and the temperature was raised to 120° C. to react overnight. After the reaction was completed, the reaction solution was concentrated under reduced pressure, and the obtained residue was separated and purified by silica gel column chromatography (eluent: system A) to obtain the product 2-(((3-bromo-4-chloro-2-fluorophenyl) Amino)methylene)-3-oxobutyric acid ethyl ester 6m (3 g, 8.23 mmol), yield: 61.56%.

MS m/z(ESI): 365.8[M+1] ⁺

Step 9

1-(7-Bromo-6-chloro-8-fluoro-4-hydroxyquinolin-3-yl)ethan-1-one

2-(((3-Bromo-4-chloro-2-fluorophenyl)amino)methylene)-3-oxobutyric acid ethyl ester 6m (1 g, 2.74 mmol) was dissolved in diphenyl ether (50 mL) During the reaction, the temperature was raised to 250°C for 1.5 hours. After the reaction was completed, the reaction solution was lowered to room temperature, poured into petroleum ether (200 mL), a solid was precipitated, filtered, and the filter cake was rinsed with petroleum ether (50 mL), the solid was collected, and dried to obtain 1-(7-bromo-6-chloro-1-(7-bromo-6-chloro). -8-Fluoro-4-hydroxyquinolin-3-yl)ethan-1-one 6n (300 mg, 941.84 μmol), yield: 34.34%.

MS m/z(ESI): 319.7[M+1] ⁺

Step 10

3-(7-Bromo-8-chloro-6-fluoro-3-methyl-1H-pyrazolo[4,3-c]quinolin-1-yl)azetidine-1-carboxylic acid tertiary Butyl ester

1-(7-Bromo-6-chloro-8-fluoro-4-hydroxyquinolin-3-yl)ethan-1-one 6n (300 mg, 941.84 μmol) was added to glacial acetic acid (5 mL) and 3-hydrazine was added tert-butyl azetidine-1-carboxylate 4d (264 mg, 1.41 mmol), the temperature was raised to 80° C. to react overnight. The reaction solution was concentrated under reduced pressure, ethyl acetate (20 mL) was added, saturated sodium bicarbonate solution was added dropwise to adjust the pH to basic, extracted with ethyl acetate (10 mL×3), washed with saturated brine (20 mL), and dried over anhydrous sodium sulfate. , filtered, dried and concentrated under reduced pressure, the obtained residue was separated and purified by silica gel column chromatography (eluent: A system) to obtain the product 3-(7-bromo-8-chloro-6-fluoro-3-methyl) -1H-pyrazolo[4,3-c]quinolin-1-yl)azetidine-1-carboxylate tert-butyl ester 6p (150 mg, 319.33 μmol), yield: 33.90%.

MS m/z(ESI): 469.0[M+1] ⁺

Step 11

3-(7-(2-((tert-butoxycarbonyl)amino)-5-fluorobenzo[d]thiazol-4-yl)-8-chloro-6-fluoro-3-methyl-1H-pyrazole tert-Butyl [4,3-c]quinolin-1-yl)azetidine-1-carboxylate

3-(7-Bromo-8-chloro-6-fluoro-3-methyl-1H-pyrazolo[4,3-c]quinolin-1-yl)azetidine-1-carboxylic acid tert-Butyl ester 6p (140mg, 298.04μmol), (2-((tert-butoxycarbonyl)amino)-5-fluorobenzo[d]thiazol-4-yl)boronic acid 6i (186.05mg, 596.08μmol), potassium phosphate (126.52 mg, 596.08 μmol) and X-PHOS Pd G2 (46.84 mg, 59.61 μmol) were dissolved in tetrahydrofuran (3 mL) and reacted at 50°C overnight. After the reaction, the filtrate was concentrated under reduced pressure, and the obtained residue was separated and purified by preparative HPLC (separation column: AKZONOBEL Kromasil; 250×21.2 mm ID; 5 μm; mobile phase A: 0.05% TFA+H2O, mobile phase B: Acetonitrile; flow rate: 20 mL/min) to give the product 3-(7-(2-((tert-butoxycarbonyl)amino)-5-fluorobenzo[d]thiazol-4-yl)-8-chloro-6- Fluoro-3-methyl-1H-pyrazolo[4,3-c]quinolin-1-yl)azetidine-1-carboxylate tert-butyl ester 6q (50 mg, 76.09 μmol), yield 25.53 %.

MS m/z(ESI): 656.8[M+1] ⁺

Step 12

4-(1-(azetidin-3-yl)-8-chloro-6-fluoro-3-methyl-1H-pyrazolo[4,3-c]quinolin-7-yl)- 5-Fluorobenzo[d]thiazol-2-amine

3-(7-(2-((tert-butoxycarbonyl)amino)-5-fluorobenzo[d]thiazol-4-yl)-8-chloro-6-fluoro-3-methyl-1H-pyridine Azolo[4,3-c]quinolin-1-yl)azetidine-1-carboxylate tert-butyl ester 6q (50 mg, 76.09 μmol) was dissolved in dichloromethane (3 mL), and 1 of hydrochloric acid was added. , 4-dioxane solution (4M, 1 mL) was reacted at room temperature for 1 hour. The reaction solution was concentrated under reduced pressure to obtain the crude product 4-(1-(azetidin-3-yl)-8-chloro-6-fluoro-3-methyl-1H-pyrazolo[4,3-c] ]quinolin-7-yl)-5-fluorobenzo[d]thiazol-2-amine 6r (34.76 mg, 76.09 μmol), yield: 100%.

MS m/z(ESI): 456.8[M+1] ⁺

step thirteen

1-(3-(7-(2-Amino-5-fluorobenzo[d]thiazol-4-yl)-8-chloro-6-fluoro-3-methyl-1H-pyrazolo[4,3 -c]quinolin-1-yl)azetidine-1-yl)prop-2-en-1-one

4-(1-(azetidin-3-yl)-8-chloro-6-fluoro-3-methyl-1H-pyrazolo[4,3-c]quinolin-7-yl) -5-Fluorobenzo[d]thiazol-2-amine 6r (33.05mg, 72.33μmol) was dissolved in dichloromethane (3mL), cooled to 0°C, triethylamine (21.96mg, 216.98μmol) and propylene were added Acid chloride (7.86 mg, 86.79 μmol), and the reaction was continued for 1 hour at 0°C. After the reaction, the reaction solution was concentrated under reduced pressure, and the obtained residue was separated and purified by preparative HPLC (separation column: AKZONOBEL Kromasil; 250×21.2 mm ID; 5 μm; mobile phase A: 0.05% TFA+H2O, mobile phase B: acetonitrile; Flow rate: 20 mL/min) to give the product 1-(3-(7-(2-amino-5-fluorobenzo[d]thiazol-4-yl)-8-chloro-6-fluoro-3-methyl- 1H-pyrazolo[4,3-c]quinolin-1-yl)azetidin-1-yl)prop-2-en-1-one 6 (5 mg, 9.16 μmol), yield: 12.67 %.

MS m/z(ESI): 512.0[M+1] ⁺

¹ H NMR(400MHz,Methanol-d4)δ9.38(d,J=4.2Hz,1H),8.54(s,1H),7.84(dd,J=8.9,5.0Hz,1H),7.15(t,J =9.2Hz,1H),6.51-6.17(m,2H),5.82(d,J=10.2Hz,1H),5.08-4.90(m,3H),4.72(m,2H),2.79(s,3H) .

Example 7

4-(1-(1-Acryloylazetidin-3-yl)-8-chloro-6-fluoro-1H-[1,2,3]triazolo[4,5-c]quinoline -7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile

first step

(3-cyano-7-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaboran-2-yl)benzo[b]thiophene-2- base) tert-butyl carbamate

(4-Bromo-3-cyano-7-fluorobenzo[b]thiophen-2-yl)carbamate tert-butyl ester 7a (300 mg, 808.14 μmol, prepared according to patent WO2021118877), 4,4,4 ',4',5,5,5',5'-Octamethyl-2,2'-bis(1,3,2-dioxaborane) (779.82 mg, 3.07 mmol), potassium acetate (237.93 mg, 2.42 mmol), bis(diphenylphosphine phenyl ether) palladium(II) dichloride (57.85 mg, 80.81 μmol) was added to 1,4-dioxane (10 mL), under argon protection, heated to 95°C, the reaction was carried out for 4 hours. After the reaction, cooled to room temperature, concentrated under reduced pressure, and prepared for liquid phase separation (separation column AKZONOBEL Kromasil; 250×21.2 mm ID; 5 μm, 20 mL/min; mobile phase A: 0.05% TFA+H2O, mobile phase B: CH3CN) , to give (3-cyano-7-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaboran-2-yl)benzo[b]thiophene- 2-yl) tert-butyl carbamate 7b (200 mg), yield: 59.17%. MS m/z(ESI): 416.9[M-1] ^-

second step

3-(7-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-8-chloro-6-fluoro-1H-[ 1,2,3]Triazolo[4,5-c]quinolin-1-yl)azetidine-1-carboxylate tert-butyl ester

(3-cyano-7-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaboran-2-yl)benzo[b]thiophene-2 -yl) tert-butyl carbamate 7b (109.91 mg, 262.76 μmol), 3-(7-bromo-8-chloro-6-fluoro-1H-[1,2,3]triazolo[4,5-c] ]quinolin-1-yl)azetidine-1-carboxylate tert-butyl ester 1e (100mg, 218.96μmol), potassium phosphate (92.95mg, 437.93μmol), chloro (2-dicyclohexylphosphino-2' ,4',6'-Triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) (17.21mg, 21.90μmol) It was added to tetrahydrofuran (10 mL), protected by argon, heated to 50°C, and reacted overnight. After the reaction, the reaction liquid was filtered through diatomaceous earth and concentrated under reduced pressure to prepare liquid phase separation (separation column AKZONOBEL Kromasil; 250 × 21.2 mm ID; 5 μm, 20 mL/min; mobile phase A: 0.05% TFA+H2O, mobile phase B: CH3CN) to give 3-(7-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-8-chloro-6 - Fluoro-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)azetidine-1-carboxylate tert-butyl ester 7c (50 mg), yield: 34.18 %.

MS m/z(ESI): 667.8[M+1] ⁺

third step

2-Amino-4-(1-(azetidin-3-yl)-8-chloro-6-fluoro-1H-[1,2,3]triazolo[4,5-c]quinoline -7-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile

3-(7-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-8-chloro-6-fluoro-1H- [1,2,3]Triazolo[4,5-c]quinolin-1-yl)azetidine-1-carboxylate tert-butyl ester 7c (50 mg, 74.84 μmol) was added to dichloromethane (3 mL ), then added 4M hydrochloric acid dioxane solution (2 mL), reacted at room temperature for 2 hours, after the reaction was completed, concentrated under reduced pressure to obtain 2-amino-4-(1-(azetidin-3-yl) -8-Chloro-6-fluoro-1H-[1,2,3]triazolo[4,5-c]quinolin-7-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile 7d (35 mg), yield: 99.96%, without purification, proceed directly to the next reaction.

MS m/z(ESI): 467.8[M+1] ⁺

the fourth step

4-(1-(1-Acryloylazetidin-3-yl)-8-chloro-6-fluoro-1H-[1,2,3]triazolo[4,5-c]quinoline -7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile

2-Amino-4-(1-(azetidin-3-yl)-8-chloro-6-fluoro-1H-[1,2,3]triazolo[4,5-c]quinoline Lin-7-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile 7d (35 mg, 74.81 μmol) was added to dichloromethane (5 mL), triethylamine (22.71 mg, 224.42 μmol) was added, and cooled To 0° C., a solution of acryloyl chloride (6.77 mg, 74.81 μmol) in dichloromethane was added dropwise, and the reaction was carried out at room temperature for 2 hours. After the reaction, concentrated under reduced pressure to prepare liquid phase separation (separation column AKZONOBEL Kromasil; 250 × 21.2 mm ID; 5 μm, 20 mL/min; mobile phase A: 0.05% TFA+H2O, mobile phase B: CH3CN) to obtain 4- (1-(1-Acryloylazetidin-3-yl)-8-chloro-6-fluoro-1H-[1,2,3]triazolo[4,5-c]quinoline-7 -yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile 7 (8 mg), yield: 19.47%.

MS m/z(ESI): 522.0[M+1] ⁺

Biological evaluation

Test Example 1. Determination of the covalent binding ability of the compound of the present invention to KRAS G12C protein

The following method was used to determine the covalent binding ability of the compounds of the present invention to recombinant human KRAS G12C protein under in vitro conditions.

The experimental procedure is briefly described as follows: Recombinant human KRAS G12C protein (aa1-169) was prepared with reaction buffer (20mM HEPES, 150mM NaCl, 1mM MgCl2, 1mM DTT) at a concentration of 4uM for use. Test compounds were dissolved in DMSO to prepare 10 mM stock solutions and subsequently diluted with reaction buffer for use. First add 1.5uL of the test compound diluted with reaction buffer (final concentration of reaction system is 3μM or 10μM) into the well, then add 23.5uL of reaction buffer and mix well, then add 25μL of 4uM recombinant human KRAS G12C protein, room temperature After 5 or 15 minutes of incubation under conditions, the reaction was stopped by the addition of 5 uL of acetic acid and the sample was transferred to the vial. An Agilent 1290/6530 instrument was used to detect the ratio of the covalent binding of the test compound to the KRAS G12C protein. The samples were subjected to liquid chromatography (XBridge Protein BEH C4,

3.5μm, 2.1mm×50mm), mobile phase A is 0.1% formic acid aqueous solution, mobile phase B is acetonitrile, mobile phase elution program is: 0 ~ 0.5 minutes, keep mobile phase A: 95%, 2.5 At 30 minutes, mobile phase A changed to 30%, and kept for 0.5 minutes, 3.1 minutes, mobile phase A changed to 95%, and kept for 1.9 minutes; flow rate: 0.5ml/min; finally use MassHunter Workstation Software Bioconfirm Version B.08.00 software The data were analyzed to obtain the covalent binding rate (Binding Rate) of the test compound to the KRAS G12C protein under the condition that the concentration of the test compound was 3 μM and incubated for 5 min, as shown in Table 1.

Table 1 Covalent binding rate of the compounds of the present invention to KRAS G12C protein

Conclusion: The compound of the present invention has a good covalent binding rate with KRAS G12C protein.

Test Example 2. The compound of the present invention inhibits the proliferation of NCI-H358 cells

The following methods were used to determine the effect of compounds of the present invention on proliferation of NCI-H358 cells. NCI-H358 cells (containing KRAS G12C mutation) were purchased from the Cell Resource Center, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, and cultured in RPMI 1640 medium containing 10% fetal bovine serum, 100 U penicillin, 100 μg/mL streptomycin and 1 mM Sodium Pyruvate middle. cell viability through

Assays were performed with the Luminescent Cell Viability Assay Kit (Promega, Cat. No. G7573).

The experimental method was operated in accordance with the steps in the kit instructions, which are briefly described as follows: the test compound was first dissolved in DMSO to prepare a 10 mM stock solution, and then diluted with culture medium to prepare a test sample. The final concentration of the compound was in the range of 1000nM-0.015nM . Cells in logarithmic growth phase were seeded into 96-well cell culture plates at a density of 800 cells per well, cultured overnight in a 37°C, 5% _CO2 incubator, followed by the addition of test compounds for 120 hours. After the incubation, 50 μL of CellTiter-Glo detection solution was added to each well, shaken for 5 minutes, and then allowed to stand for 10 minutes. Then, the luminescence value of each well of the sample was read on a microplate reader using Luminescence mode. By comparing with the value of the control group (0.3% DMSO), the percentage inhibition rate of the compound at each concentration point was calculated, and then a nonlinear regression analysis was performed in the GraphPad Prism 5 software with the compound concentration logarithm-inhibition rate to obtain the compound inhibiting cell proliferation. The _IC50 values are shown in Table 2.

Table 2 IC ₅₀ data of the compounds of the present invention on NCI-H358 cell proliferation inhibition

Conclusion: The compounds of the present invention have a good proliferation inhibition effect on NCI-H358 (human non-small cell lung cancer) cells.

Test Example 3. Determination of the Inhibitory Activity of the Compounds of the Invention on p-ERK1/2 in NCI-H358 Cells

The following method was used to determine the inhibitory activity of the compounds of the present invention on p-ERK1/2 in NCI-H358 cells. This method uses the Advanced phospho-ERK1/2 (Thr202/tyr204) kit (Cat. No. 64AERPEH) from Cisbio. For detailed experimental operations, please refer to the kit instructions. NCI-H358 cells (containing KRAS G12C mutation) were purchased from the Cell Resource Center of Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences.

The experimental procedure is briefly described as follows: NCI-H358 cells were cultured in complete RPMI 1640 medium containing 10% fetal bovine serum, 100 U penicillin, 100 μg/mL streptomycin and 1 mM Sodium Pyruvate. NCI-H358 cells were plated in 96-well plates at 30,000 cells per well, and the medium was complete medium, and cultured overnight in a 37°C, 5% CO ₂ incubator. The test compound was dissolved in DMSO to prepare a 10mM stock solution, then diluted with RPMI 1640 basal medium, and 90 μL of RPMI 1640 basal medium containing the corresponding concentration of the test compound was added to each well. The concentration range is 1000nM-0.015nM, placed in a cell incubator for 3 hours and 40 minutes. Subsequently, 10 μL of hEGF prepared in RPMI 1640 basal medium (purchased from Roche, Cat. No. 11376454001) was added to a final concentration of 5 nM and incubated in an incubator for 20 minutes. Discard the cell supernatant, wash the cells with ice-bathed PBS, then add 45 μL of 1×cell phospho/total protein lysis buffer (component of the Advanced phospho-ERK1/2 kit) to each well for lysis, and place the 96-well plate on ice Lysis was carried out for half an hour, and then the lysate was detected according to the instructions of the Advanced phospho-ERK1/2 (Thr202/tyr204) kit. Finally, under the excitation wavelength of 304nM, the fluorescence intensity of each well at 620nM and 665nM was measured on a microplate reader in TF-FRET mode, and the fluorescence intensity ratio of 665/620 in each well was calculated. By comparing the fluorescence intensity ratio with the control group (0.1% DMSO), the percentage inhibition rate of the test compound at each concentration was calculated, and the numerical-inhibition rate was subjected to nonlinear regression analysis with the test compound concentration by GraphPad Prism 5 software. , to obtain the _IC50 value of the compound.

Conclusion: The compounds of the present invention have a good proliferation inhibitory effect on p-ERK1/2 in NCI-H358 cells.

Claims (16)

1. A compound represented by the general formula (I) or a stereoisomer, tautomer or a pharmaceutically acceptable salt thereof:

   

   in:

   E is selected from

   

   G is a 4- to 12-membered heterocyclic group containing 1-2 nitrogen atoms, preferably a 4-membered heterocyclic group, wherein the heterocyclic group is optionally further substituted by one or more R ^c ;

   L is a chemical bond or a C ₁ -C ₆ alkylene group; wherein the alkylene group is optionally further substituted by one or more substituents selected from alkyl, halogen or hydroxyl; preferably, L is a chemical bond or -CH ₂ -, -CH ₂ CH ₂ - or -CH(CH ₃ )-; more preferably, L is a chemical bond;

   X and Y are each independently selected from N or CR ^f ;

   Ring A is selected from the group:

   

   The condition is that when ring A is selected from

   

   When , X is selected from N, Y is selected from CR ^f , the carbon atom of G is connected with ring A;

   When ring A is selected from

   

   , -LR ⁴ does not exist;

   W is selected from N or CR ^d ;

   Z is selected from N or CR ^e ;

   Ring B is selected from 5-6 membered heteroaryl;

   Ring C is selected from 5-6 membered heteroaryl;

   ^Ra is selected from hydrogen atoms or fluorine;

   R ^b is selected from hydrogen atom, -CH ₂ F, -CHF ₂ ,

   

   R ^c is the same or different, and each is independently selected from a hydrogen atom, an alkyl group, a halogen, a nitro group, a cyano group, a cycloalkyl group, a heterocyclic group, an aryl group, a heteroaryl group, =O, -OR ⁵ , -C( O)R ⁵ , -C(O)OR ⁵ , -NHC(O)R ⁵ , -NHC(O)OR ⁵ , -NR ⁶ R ⁷ , -C(O)NR ⁶ R ⁷ , -CH ₂ NHC ( O)OR ⁵ , -CH ₂ NR ⁶ R ⁷ or -S(O) _r R ⁵ , wherein said alkyl, cycloalkyl, heterocyclyl, aryl or heteroaryl is optionally further modified by one or more Multiple selected from alkyl, halogen, nitro, cyano, cycloalkyl, heterocyclyl, aryl, heteroaryl, =O, ^-OR5 , -C(O) ^R5 , -C(O) OR ⁵ , -NHC(O)R ⁵ , -NHC(O)OR ⁵ , -NR ⁶ R ⁷ , -C(O)NR ⁶ R ⁷ , -CH ₂ NHC(O)OR ⁵ , -CH ₂ NR ⁶ R ⁷ or -S(O) _r R ⁵ is substituted by the substituent;

   R ^d and R ^e are the same or different, each independently selected from hydrogen atom, halogen, alkyl, alkoxy, cyano, nitro, amino, hydroxyl, haloalkyl or haloalkoxy, preferably cyano;

   R ^{f is} selected from hydrogen atom, halogen, alkyl or alkoxy; wherein said alkyl or alkoxy is optionally further selected by one or more of halogen, hydroxy, cyano, alkyl or alkoxy substituted by the substituent; R ^f is preferably halogen, more preferably fluorine or chlorine;

   R ¹ is selected from hydrogen atom, halogen, alkyl or alkoxy; wherein said alkyl or alkoxy is optionally further selected by one or more of halogen, hydroxyl, cyano, alkyl or alkoxy substituted by the substituent; R ¹ is preferably a hydrogen atom;

   R ² is selected from aryl or heteroaryl, wherein said aryl or heteroaryl is optionally further substituted with one or more ^RA ;

   R ^A is each independently selected from alkyl, halogen, nitro, cyano, cycloalkyl, heterocyclyl, aryl, heteroaryl, =O, ^-OR5 , -C(O) ^R5 , -C (O)OR ⁵ , -NHC(O)R ⁵ , -NHC(O)OR ⁵ , -NR ⁶ R ⁷ , -C(O)NR ⁶ R ⁷ , -CH ₂ NHC(O)OR ⁵ , -CH ₂ NR ⁶ R ⁷ or -S(O) _r R ⁵ ; wherein said alkyl, cycloalkyl, heterocyclyl, aryl or heteroaryl is optionally further selected from one or more of alkyl, halogen, nitro, cyano, cycloalkyl, heterocyclyl, aryl, heteroaryl, =O, ^-OR5 , -C(O)R5, -C(O ^{) OR5} , -NHC(O )R ⁵ -NHC(O)OR ⁵ , -NR ⁶ R ⁷ , -C(O)NR ⁶ R ⁷ , -CH ₂ NHC(O)OR ⁵ , -CH ₂ NR ⁶ R ⁷ or -S(O) _r R ⁵ is substituted by the substituent;

   The condition is that when ring A is selected from

   

   , R ² is not selected from

   

   R ³ is selected from hydrogen atom, halogen, alkyl, alkoxy, cyano, haloalkyl or haloalkoxy, preferably hydrogen;

   R ⁴ is selected from a hydrogen atom, an aryl group or a heteroaryl group; wherein said aryl or heteroaryl group is optionally further substituted by one or more R ^B ; R ⁴ is preferably a heteroaryl group.

   Each R ^B is independently selected from alkyl, halogen, nitro, cyano, cycloalkyl, heterocyclyl, aryl, heteroaryl, =O, ^-OR5 , -C(O) ^R5 , -C (O)OR ⁵ , -NHC(O)R ⁵ -NHC(O)OR ⁵ , -NR ⁶ R ⁷ , -C(O)NR ⁶ R ⁷ , -CH ₂ NHC(O)OR ⁵ , -CH ₂ NR ⁶ R ⁷ or -S(O) _r R ⁵ ; wherein said alkyl, cycloalkyl, heterocyclyl, aryl or heteroaryl is optionally further , nitro, cyano, cycloalkyl, heterocyclyl, aryl, heteroaryl, =O, -OR ⁵ , -C(O)R ⁵ , -C(O)OR ⁵ , -NHC(O) R ⁵ -NHC(O)OR ⁵ , -NR ⁶ R ⁷ , -C(O)NR ⁶ R ⁷ , -CH ₂ NHC(O)OR ⁵ , -CH ₂ NR ⁶ R ⁷ or -S(O) _r Replaced by the substituent of R ⁵ ;

   R ⁵ is selected from a hydrogen atom, an alkyl group, a cycloalkyl group, a heterocyclyl group, an aryl group or a heteroaryl group, wherein said alkyl group, cycloalkyl group, heterocyclyl group, aryl group or heteroaryl group is optionally further One or more selected from hydroxy, halogen, nitro, cyano, alkyl, alkoxy, haloalkyl, haloalkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl, =O, - C(O)R ⁸ , -C(O)OR ⁸ , -OC(O)R ⁸ , -NR ⁹ R ¹⁰ , -C(O)NR ⁹ R ¹⁰ , -SO ₂ NR ⁹ R ¹⁰ or -NR ⁹ Substituents of C(O)R ¹⁰ are substituted;

   R ⁶ and R ⁷ are each independently selected from a hydrogen atom, a hydroxyl group, a halogen, an alkyl group, an alkoxy group, a cycloalkyl group, a heterocyclic group, an aryl group or a heteroaryl group, wherein the alkyl group, alkoxy group, A cycloalkyl, heterocyclyl, aryl or heteroaryl group is optionally further selected from one or more groups selected from hydroxy, halogen, nitro, cyano, alkyl, alkoxy, cycloalkyl, heterocyclyl, Aryl, heteroaryl, =O, -C(O)R ⁸ , -C(O)OR ⁸ , -OC(O)R ⁸ , -NR ⁹ R ¹⁰ , -C(O)NR ⁹ R ¹⁰ , -SO ₂ NR ⁹ R ¹⁰ or -NR ⁹ C(O)R ¹⁰ substituent;

   Alternatively, R ⁶ and R ⁷ together with the atoms to which they are attached form a 4- to 8-membered heterocyclyl group containing one or more N, O or S(O) _r within the 4- to 8-membered heterocyclyl group, and The 4-8 membered heterocyclic group is optionally further selected from one or more groups selected from hydroxyl, halogen, nitro, cyano, alkyl, alkoxy, cycloalkyl, heterocyclic, aryl, heterocyclic Aryl, =O, -C(O)R ⁸ , -C(O)OR ⁸ , -OC(O)R ⁸ , -NR ⁹ R ¹⁰ , -C(O)NR ⁹ R ¹⁰ , -SO ₂ NR ⁹ R ¹⁰ or -NR ⁹ C(O) R ¹⁰ is substituted with a substituent;

   R ⁸ , R ⁹ and R ¹⁰ are each independently selected from a hydrogen atom, an alkyl group, an amino group, a cycloalkyl group, a heterocyclic group, an aryl group or a heteroaryl group, wherein said alkyl group, cycloalkyl group, heterocyclic group , aryl or heteroaryl are optionally further selected from one or more groups selected from hydroxy, halogen, nitro, amino, cyano, alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl substituted by substituents of carboxylate, carboxyl or carboxylate;

   R ¹¹ is selected from hydrogen atom, halogen, alkyl, alkoxy, cyano, nitro, amino, hydroxyl, haloalkyl, haloalkoxy or =O;

   R ¹² is selected from a hydrogen atom, an alkyl group, -C(O)R ¹³ or -S(O) ₂ R ¹³ ;

   R ¹³ is selected from alkyl, preferably methyl;

   n is each independently selected from 0, 1 or 2;

   r is each independently 0, 1 or 2.
2. The compound according to claim 1 or a stereoisomer, tautomer or a pharmaceutically acceptable salt thereof, which is a compound represented by the general formula (II) or a stereoisomer or tautomer thereof body or its pharmaceutically acceptable salt:

   

   in:

   Ring B, R ¹ to R ³ , X, Y, G, E and n are as defined in claim 1 .
3. The compound according to claim 1 or 2 or a stereoisomer, tautomer or pharmaceutically acceptable salt thereof, wherein:

   -G-E are selected from:

   

   R ^c are the same or different, each independently selected from hydrogen atom, halogen, alkyl or alkoxy, preferably alkyl, more preferably methyl;

   m is selected from 0, 1, 2, 3 or 4;

   E is as defined in claim 1 .
4. The compound according to any one of claims 1 to 3, or a stereoisomer, tautomer or pharmaceutically acceptable salt thereof, wherein E is selected from:

   
5. The compound according to any one of claims 1-4 or a stereoisomer, tautomer or a pharmaceutically acceptable salt thereof, wherein X, Y are each independently selected from CR ^f ; R ^{f is} selected from hydrogen atom, halogen, alkyl, alkoxy, haloalkyl or haloalkoxy, R ^f is preferably halogen, more preferably fluorine or chlorine.
6. The compound according to any one of claims 1-5 or a stereoisomer, tautomer or a pharmaceutically acceptable salt thereof, wherein R ¹ is selected from hydrogen atom, halogen, alkyl, alkoxy, haloalkyl or haloalkoxy, R ¹ is preferably a hydrogen atom.
7. The compound according to any one of claims 1-6 or a stereoisomer, tautomer or pharmaceutically acceptable salt thereof, wherein:

   R ² is selected from phenyl, naphthyl, pyridyl, benzothiazolyl or benzopyrazolyl, wherein said phenyl, naphthyl, pyridyl, benzothiazolyl or benzopyrazolyl is optionally further replaced by one or more ^RAs ;

   R ^A is each independently selected from halogen, hydroxy, alkyl, alkoxy, cycloalkyl or -NR ⁶ R ⁷ , wherein said alkyl or alkoxy is optionally further selected by one or more selected from halogen or -Substituents of NR ⁶ R ⁷ are substituted; wherein the halogen is preferably fluorine;

   R ⁶ and R ⁷ are each independently selected from a hydrogen atom or an alkyl group, wherein the alkyl group is preferably a methyl group.
8. ^A compound according to any one of claims 1-7 or a stereoisomer, tautomer or pharmaceutically acceptable salt thereof, wherein R is selected from:

   

   
9. The compound of any one of claims 1-8, or a stereoisomer, tautomer, or a pharmaceutically acceptable salt thereof, wherein ^R3 is selected from a hydrogen atom.
10. A compound according to any one of claims 1-9 or a stereoisomer, tautomer or pharmaceutically acceptable salt thereof, wherein Ring B is selected from:

    
11. The compound according to any one of claims 1-10 or a stereoisomer, tautomer or a pharmaceutically acceptable salt thereof, wherein the compound is:

    
12. A preparation method of a compound of general formula (II) according to any one of claims 2-11 or a stereoisomer, tautomer or a pharmaceutically acceptable salt thereof, the method comprising the steps of:

    

    The compound of general formula (IIA) is reacted with the compound of general formula (IIB) under basic conditions, and the protecting group is optionally further removed to obtain the compound of general formula (II);

    in:

    X ¹ is a leaving group, preferably selected from chlorine, bromine, iodine, OMs or OTs, more preferably chlorine;

    Ring B, R ¹ to R ³ , X, Y, G, E and n are as defined in claim 2 .
13. A pharmaceutical composition containing an effective dose of a compound according to any one of claims 1 to 11 or a stereoisomer, tautomer or a pharmaceutically acceptable salt thereof, and optionally a Pharmaceutically acceptable carriers, excipients or combinations thereof.
14. The compound according to any one of claims 1 to 11 or a stereoisomer, a tautomer or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 13 in the preparation of K-Ras Use in a GTPase inhibitor, wherein the K-Ras GTPase inhibitor is preferably a KRAS G12C inhibitor.
15. The compound according to any one of claims 1 to 11 or a stereoisomer, tautomer or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 13 in preparation for use in therapy Use in medicine for diseases mediated by KRAS mutation, wherein said disease mediated by KRAS mutation is preferably selected from cancer, wherein said cancer is preferably selected from pancreatic cancer, colorectal cancer, lung cancer, multiple myeloma , uterine cancer, bile duct cancer, gastric cancer, bladder cancer, diffuse large B cell lymphoma, rhabdomyosarcoma, skin squamous cell carcinoma, cervical cancer, testicular germ cell cancer, particularly preferably, the cancer is preferably pancreatic cancer, colon cancer Rectal cancer and lung cancer, wherein said KRAS mutation is preferably KRAS G12C mutation.
16. The compound according to any one of claims 1 to 11 or a stereoisomer, tautomer or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 13 in preparation for use in therapy Use in the medicament of cancer, wherein said cancer is preferably selected from pancreatic cancer, colorectal cancer, lung cancer, multiple myeloma, uterine cancer, bile duct cancer, gastric cancer, bladder cancer, diffuse large B-cell lymphoma, rhabdomyosarcoma , skin squamous cell carcinoma, cervical cancer, testicular germ cell carcinoma, more preferably selected from pancreatic cancer, colorectal cancer and lung cancer; wherein the lung cancer is preferably non-small cell lung cancer.