CN116390923A - Heterocyclic derivative and preparation method and application thereof - Google Patents

Abstract

The invention relates to heterocyclic derivatives, a preparation method and medical application thereof. In particular, the invention relates to heterocyclic derivatives represented by the general formula (I), a preparation method and pharmaceutically acceptable salts thereof, and application of the heterocyclic derivatives as therapeutic agents, particularly as K-Ras GTPase inhibitors, wherein each substituent group in the general formula (I) is defined as the specification.

Description

Heterocyclic derivative and preparation method and application thereof

The present application claims priority from the following chinese patent applications:

1) The invention relates to a heterocyclic derivative, a preparation method and application thereof, which are submitted to Chinese patent application 202010847639.9 of Chinese patent office in 8/21/2020;

2) The invention is named as heterocyclic derivatives, a preparation method and application thereof, and the heterocyclic derivatives are submitted to China patent application 202110323635.3 of China patent office on the day 26 of 3 months of 2021;

3) The invention is named as heterocyclic derivatives, a preparation method and application thereof, which are submitted to China patent application 202110569537.X of China patent office in 5-25 of 2021;

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

Technical Field

The invention relates to a heterocyclic derivative, a preparation method thereof, a pharmaceutical composition containing the derivative and application of the heterocyclic derivative as a therapeutic agent, in particular to an inhibitor of K-Ras GTPase.

Background

RAS represents a closely related group of monomeric globular proteins (21 kDa molecular weight) with 189 amino acids and which are associated with the plasma membrane and bind GDP or GTP. Under normal developmental or physiological conditions, the RAS is activated by receiving growth factors and various other extracellular signals, responsible for regulating functions such as cell growth, survival, migration and differentiation. RAS functions as a molecular switch, the on/off state of the RAS protein is determined by nucleotide binding, the active signaling conformation binds GTP, and the inactive conformation binds GDP. When the RAS contains bound GDP, it is in a dormant or quiescent or off state and is "inactive". When cells are exposed to certain growth promoting stimuli in response, the RAS is induced and converts the bound GDP to GTP. As GTP is bound, the RAS is "on" and is able to interact with and activate other proteins (its "downstream targets"). RAS proteins themselves have a very low inherent ability to hydrolyze GTP back to GDP and thereby turn themselves into an off state. Conversion of the RAS to shut down requires exogenous proteins called Gtpase Activating Proteins (GAPs) that interact with the RAS and greatly promote the conversion of GTP to GDP. Any mutation in the 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 produce a prolonged signal to the cell that signals it to continue growth and division. These signals may therefore cause cell growth and division, and overactivated RAS signaling may ultimately lead to cancer.

Structurally, RAS proteins contain a G domain responsible for enzymatic activity of the ras— guanine nucleotide binding and hydrolysis (gtpase reaction). It also includes a C-terminal extension called CAAX box, which can be post-translationally modified and targets the protein to a membrane. The G domain is approximately 21-25kDa in size and contains a phosphate binding loop (P-loop). The P-loop represents the pocket in the protein that binds the nucleotide and this is a rigid part of the domain with conserved amino acid residues that are necessary for nucleotide binding and hydrolysis (glycine 12, threonine 26 and lysine 16). The G domain also contains so-called switch I regions (residues 30-40) and switch II regions (residues 60-76), which are both dynamic parts of the protein, often denoted as "spring-loaded" mechanisms due to the ability of the dynamic parts to switch between resting and loaded states. The main interaction is the hydrogen bond formed by threonine-35 and glycine-60 with the gamma-phosphate of GTP, which maintains their active conformation in switch I and switch II, respectively. After hydrolysis of GTP and release of phosphate, both relax to an inactive GDP conformation.

Among RAS family members, oncogenic mutations are most common in KRAS (85%), while NRAS (12%) and HRAS (3%) are less common. KRAS mutations are prevalent in three major deadly 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 multiple myeloma, uterine cancer, cholangiocarcinoma, gastric cancer, bladder cancer, diffuse large B-cell lymphoma, rhabdomyosarcoma, cutaneous squamous cell carcinoma, cervical cancer, testicular germ cell carcinoma, etc., whereas KRAS mutations are rarely found (< 2%) in breast, ovarian and brain cancers. 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 increase in the frequency of specific allelic mutations comes mostly from classical smoking-induced classical mutations (G: C to T: A substitutions), resulting in KRAS G12C (GGT to TGT) and G12V (GGT to GTT) mutations.

Large genomics studies indicate that lung cancer KRAS mutations, including G12C, are mutually exclusive with other known driving oncogenic mutations in NSCLC (including EGFR, ALK, ROS1, RET, and BRAF), indicating the uniqueness of KRAS mutations in lung cancer. While at the same time KRAS mutations often coincide with certain co-mutations, such as STK11, KEAP1 and TP53, which in cooperation with the mutated RAS transform the cells into highly malignant and invasive tumor cells.

Three RAS oncogenes constitute the most frequently mutated gene family in human cancers. It is disappointing that despite thirty years of research efforts, there is still no clinically effective anti-RAS therapy, and targeting the gene using small molecules is a challenge. Accordingly, there is an urgent need in the art for small molecules for targeting the RAS (e.g., K-RAS, H-RAS, and/or N-RAS) and using the same to treat a variety of diseases, such as cancer.

At present, the clinical development of KRAS inhibitor at home and abroad is very competitive, wherein KRAS enzyme inhibitor MRTX-849 developed by Mirati Therapeutics Inc company already enters clinical stage two for preventing and treating diseases such as advanced solid tumor, metastatic colorectal cancer, metastatic non-small cell lung cancer and the like. There are other KRAS inhibitors under investigation including AMG-510 (Amgen Inc, phase 2) and MRTX1257 (Mirati Therapeutics Inc, found). Early clinical studies showed that KRAS inhibitors can control and alleviate disease progression in non-small cell lung cancer patients and can reduce tumor size in patients with advanced lung cancer and colorectal cancer. A series of KRAS inhibitor patent applications have been published, including WO2020047192, WO2019099524 and WO2018217651, etc., showing that there has been some progress in the research and use of KRAS inhibitors, however, existing KRAS inhibitors are still not satisfactory in terms of effectiveness and safety, and there is still a great room for improvement, and there is still a need to continue to research and develop new KRAS inhibitors.

Disclosure of Invention

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

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

wherein:

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 with one or more R ^c Substituted;

l is a bond or C ₁ -C ₆ An alkylene group; wherein said alkylene is optionally further substituted with one or more substituents selected from alkyl, halogen or hydroxy; preferably, L is a bond, -CH ₂ -、-CH ₂ CH ₂ -or-CH (CH) ₃ ) -; more preferably, L is a bond;

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

Ring a is selected from the group consisting of:

with the proviso 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 attached to ring A;

when ring A is selected from

When, -L-R ⁴ Absence of;

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;

R ^a selected from hydrogen atoms or fluorine;

R ^b selected from hydrogen atoms, -CH ₂ F、-CHF ₂ 、

R ^c The same OR different are each independently selected from hydrogen atom, alkyl, halogen, 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 R ⁵ Wherein said alkyl, cycloalkyl, heterocyclyl, aryl or heteroaryl is optionally further substituted with one or more substituents selected from alkyl, halo, 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 R ⁵ Is substituted by a substituent of (2);

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

R ^f selected from a hydrogen atom, halogen, alkyl or alkoxy; wherein said alkyl or alkoxy is optionally further substituted with one or more substituents selected from halogen, hydroxy, cyano, alkyl or alkoxy; r is R ^f Preferably halogen, more preferably fluorine or chlorine;

R ¹ selected from a hydrogen atom, halogen, alkyl or alkoxy; wherein said alkyl or alkoxy is optionally further substituted with one or more substituents selected from halogen, hydroxy, cyano, alkyl or alkoxy; r is R ¹ Preferably a hydrogen atom;

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

R ^A each independently selected from alkyl, halo, 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 R ⁵ The method comprises the steps of carrying out a first treatment on the surface of the Wherein said alkyl, cycloalkyl, heterocyclyl, aryl orThe heteroaryl group is optionally further substituted with one OR more groups selected from alkyl, halo, 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 R ⁵ Is substituted by a substituent of (2);

with the proviso that when ring A is selected from

When R is ² Not selected from

R ³ Selected from a hydrogen atom, halogen, alkyl, alkoxy, cyano, haloalkyl or haloalkoxy, preferably a hydrogen atom;

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

R ^B each independently selected from alkyl, halo, 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 R ⁵ The method comprises the steps of carrying out a first treatment on the surface of the Wherein said alkyl, cycloalkyl, heterocyclyl, arylOR heteroaryl optionally further substituted with one OR more substituents selected from alkyl, halogen, 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 R ⁵ Is substituted by a substituent of (2);

R ⁵ each independently selected from a hydrogen atom, an alkyl group, a cycloalkyl group, a heterocyclic group, an aryl group, or a heteroaryl group, wherein the alkyl group, cycloalkyl group, heterocyclic group, aryl group, or heteroaryl group is optionally further substituted with one or more groups selected from hydroxy, halogen, nitro, cyano, alkyl, alkoxy, haloalkyl, haloalkoxy, cycloalkyl group, heterocyclic group, aryl group, 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 Is substituted by a substituent of (2);

R ⁶ and R is ⁷ Each independently selected from a hydrogen atom, hydroxy, halogen, alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl or heteroaryl, wherein said alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl or heteroaryl is optionally further substituted with one or more substituents 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 Is substituted by a substituent of (2);

alternatively, R ⁶ And R is ⁷ Together with the atoms to which they are attached form a 4-8 membered heterocyclic group, wherein the 4-8 membered heterocyclic group contains one or more of N, O or S (O) _r And said 4-8 membered heterocyclyl is optionally further substituted with one or more substituents 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 Is substituted by a substituent of (2);

R ⁸ 、R ⁹ and R is ¹⁰ Each independently selected from the group consisting of hydrogen, alkyl, amino, cycloalkyl, heterocyclyl, aryl, or heteroaryl, wherein said alkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl is optionally further substituted with one or more substituents selected from the group consisting of hydroxy, halogen, nitro, amino, cyano, alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl, carboxyl, or carboxylate;

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

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

R ¹³ Selected from alkyl groups, preferably methyl;

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

r are each independently 0, 1 or 2.

In a preferred embodiment of the present invention, the compound of formula (I) or a stereoisomer, tautomer or pharmaceutically acceptable salt thereof is a compound of formula (II):

wherein:

ring B, R ¹ ～R ³ The definitions of X, Y, G, E and n are as described in the general formula (I).

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

-G-E is selected from:

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

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

e is defined as in formula (I).

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

in a preferred embodiment of the invention, the compounds of the general formula (I) or (II) or stereoisomers, tautomers or pharmaceutically acceptable salts thereof, wherein X, Y are each independently selected from CR ^f ；R ^f Selected from hydrogen atoms, halogen, alkyl groups, alkoxy groupsHaloalkyl or haloalkoxy, R ^f More preferably halogen, particularly preferably fluorine or chlorine.

In a preferred embodiment of the invention, for a compound of formula (I) or (II) or a stereoisomer, tautomer or pharmaceutically acceptable salt thereof, wherein R ¹ Selected from hydrogen atom, halogen, alkyl, alkoxy, haloalkyl or haloalkoxy, R ¹ More preferably a hydrogen atom.

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

R ² Selected from phenyl, naphthyl, pyridinyl, benzothiazolyl, or benzopyrazolyl, wherein said phenyl, naphthyl, pyridinyl, benzothiazolyl, or benzopyrazolyl is optionally further substituted with one or more R ^A Substitution;

R ^A each independently selected from halogen, hydroxy, alkyl, alkoxy, cycloalkyl or-NR ⁶ R ⁷ Wherein said alkyl or alkoxy is optionally further substituted with one or more groups selected from halogen or-NR ⁶ R ⁷ Is substituted by a substituent of (2); wherein said halogen is preferably fluorine;

R ⁶ and R is ⁷ 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 invention, for a compound of formula (I) or (II) or a stereoisomer, tautomer or pharmaceutically acceptable salt thereof, wherein R ² Selected from:

in a preferred embodiment of the invention, for a compound of formula (II) or a stereoisomer, tautomer or pharmaceutically acceptable salt thereof, wherein R ³ Selected from hydrogen atoms.

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

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

Or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof.

Note that: if there is a difference between a structural formula and the name given to the structural formula, the structural formula is subject to.

Further, the present invention provides a process for the preparation of a compound of formula (II), or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, comprising the steps of:

reacting a compound of formula (IIA) with a compound of formula (IIB) under basic conditions, optionally further deprotecting to give a compound of formula (II);

wherein:

X ¹ as leaving group, preferably chloro, bromo, iodo, OMs or OTs; more preferably chlorine;

ring B, R ¹ ～R ³ The definitions of X, Y, G, E and n are as described in formula (II).

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

In another aspect, the invention provides a method of inhibiting a K-Ras GTPase, wherein the method comprises administering to a subject (including patients and healthy subjects) in need thereof a pharmaceutical composition comprising an effective amount of a compound of formula (I) or (II), or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, and optionally a pharmaceutically acceptable carrier, excipient, or combination thereof, wherein the K-Ras GTPase is preferably KRAS G12C.

The present invention also provides the use of a compound of formula (I) or (II) or a stereoisomer, tautomer or pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, for the manufacture of a medicament for the treatment of a disease mediated by a KRAS mutation, wherein the disease mediated by a KRAS mutation is preferably selected from cancer, wherein the cancer is preferably selected from pancreatic cancer, colorectal cancer, lung cancer, multiple myeloma, uterine cancer, cholangiocarcinoma, gastric cancer, bladder cancer, diffuse large B-cell lymphoma, rhabdomyosarcoma, cutaneous 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.

In another aspect, the invention provides the use of a compound of formula (I) or (II), or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, in the preparation of 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 for preventing and/or treating KRAS mutation-mediated diseases, comprising administering to a patient a therapeutically effective amount of a compound of formula (I) or (II), or a tautomer, a meso, a racemate, an enantiomer, a diastereomer, or a mixture thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the same, wherein the KRAS mutation is preferably a KRAS G12C mutation.

The present invention also provides the use of a compound of formula (I) or (II) or a stereoisomer, tautomer or pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, for the manufacture of a medicament for the treatment of cancer, wherein the cancer is preferably selected from pancreatic cancer, colorectal cancer, lung cancer, multiple myeloma, uterine cancer, cholangiocarcinoma, gastric cancer, bladder cancer, diffuse large B-cell lymphoma, rhabdomyosarcoma, cutaneous 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 may be administered topically, orally, transdermally, rectally, vaginally, parenterally, intranasally, intrapulmonary, intraocular, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intradermal, intraperitoneal, subcutaneous, subcuticular or by inhalation. Pharmaceutical compositions containing the active ingredient may be in a form suitable for oral administration, for example, as tablets, troches, 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 non-toxic pharmaceutically acceptable excipients which are suitable for the manufacture of tablets.

The formulations of the present invention are suitably presented in unit dosage form and may be prepared by any method well known in the pharmaceutical arts. The amount of active ingredient that can be combined with the carrier material to produce a single dosage form can vary depending upon the host treated and the particular mode of administration. The amount of active ingredient that can be combined with a carrier material to produce a single dosage form generally refers to the amount of compound that is capable of producing a therapeutic effect.

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

When the compounds of the invention are administered to humans and animals in the form of a medicament, the compounds may be provided alone or in the form of a pharmaceutical composition, which may contain the active ingredient in combination with a pharmaceutically acceptable carrier, for example 0.1% to 99.5% (more preferably 0.5% to 90%) of the active ingredient.

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 its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose, and cellulose acetate; (4) powdered 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) buffering agents such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) ethanol; (20) phosphate buffer solution; (21) Cyclodextrins, e.g., targeting ligands attached to nanoparticles, e.g., accursinTM; and (22) other non-toxic compatible substances used 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, cysteamine hydrochloride, sodium bisulfate, 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 chelators such as citric acid, ethylenediamine tetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid and the like. Solid dosage forms (e.g., capsules, dragees, powders, granules and the like) may include one or more pharmaceutically acceptable carriers, such as sodium citrate or dicalcium phosphate, and/or any of the following: (1) Fillers or extenders, such as starch, lactose, sucrose, glucose, mannitol, and/or silicic acid; (2) Binders, such as carboxymethyl cellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose, and/or acacia; (3) humectants, such as glycerin; (4) Disintegrants, for example agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate; (5) dissolution retarders, such as paraffin; (6) an absorption accelerator, such as a quaternary ammonium compound; (7) Humectants, such as cetyl alcohol and glycerol monostearate; (8) absorbents 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) a colorant. Liquid dosage forms may include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active ingredient, the liquid dosage form may contain inert diluents commonly used in the art, such as water or other solvents; solubilizing agents and emulsifiers, for example, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1, 3-butylene glycol, oils (in particular cottonseed, groundnut, corn, germ, olive, castor, and sesame oils), glycerol, tetrahydrofuryl alcohol, polyethylene glycols 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-agar, and tragacanth, and mixtures thereof.

In addition to the active compounds, ointments, pastes, creams and gels may contain excipients such as animal and vegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicic acid, talc and zinc oxide, or mixtures thereof.

In addition to the active compounds, the powders and sprays can also contain excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicates and polyamide powder or mixtures of these substances. The spray may contain other conventional propellants such as chlorofluorohydrocarbons, and volatile unsubstituted hydrocarbons such as butane and propane.

Detailed description of the invention

Unless stated to the contrary, some of the 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 C ₁ -C ₂₀ Straight chain or branched aliphatic hydrocarbon groups. Preferably C ₁ -C ₁₀ Alkyl, more preferably C ₁ -C ₆ Alkyl, or C ₁ -C ₄ An 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-dimethylpropyl, 1, 2-dimethylpropyl, 2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1, 2-trimethylpropyl, 1-dimethylbutyl, 1, 2-dimethylbutyl, 2-dimethylbutyl, 1, 3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2, 3-dimethylbutyl, and the like. Alkyl groups may 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, vinyl, 1-propenyl, 2-propenyl, 1-, 2-or 3-butenyl, and the like. Preferably is C ₂ -C ₁₀ Alkenyl groups of (C) are more preferred ₂ -C ₆ Alkenyl groups, most preferably C ₂ -C ₄ Alkenyl groups. Alkenyl groups may be optionally substituted or unsubstituted.

"alkynyl" refers to an aliphatic hydrocarbon group containing one carbon-carbon triple bond, which may be straight or branched. Preferably is C ₂ -C ₁₀ More preferably C ₂ -C ₆ Alkynyl, most preferably C ₂ -C ₄ 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 may be substituted or unsubstituted.

"cycloalkyl" refers to saturated or partially saturated monocyclic, fused, bridged, and spiro carbocycles. Preferably C ₃ -C ₁₂ Cycloalkyl, more preferably C ₃ -C ₈ Cycloalkyl, most preferably C ₃ -C ₆ Cycloalkyl groups. Examples of monocyclic cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatrienyl, cyclooctyl, and the like, with cyclopropyl, cyclohexenyl being preferred. Cycloalkyl groups may be optionally substituted or unsubstituted.

"spirocycloalkyl" refers to a 5 to 18 membered, two or more cyclic structure, and monocyclic polycyclic groups sharing one carbon atom (called spiro atom) with each other, containing 1 or more double bonds within the ring, but no ring has a completely conjugated pi-electron aromatic system. The spirocycloalkyl group is preferably a 6-to 14-membered spirocycloalkyl group, more preferably a 7-to 10-membered spirocycloalkyl group. The spirocycloalkyl group is classified into a single spiro group, a double spiro group or a multiple spirocycloalkyl group according to the number of common spiro atoms between rings, preferably single spiro group and double spirocycloalkyl group, preferably 4-membered/5-membered, 4-membered/6-membered, 5-membered/5-membered or 5-membered/6-membered. 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 ring alkyl" refers to an all-carbon polycyclic group having 5 to 18 members, two or more cyclic structures sharing a pair of carbon atoms with each other, one or more of the rings may contain one or more double bonds, but none of the rings has an aromatic system with fully conjugated pi electrons, preferably a 6 to 12 member fused ring alkyl group, more preferably a 7 to 10 member fused ring alkyl group. The number of constituent rings may be classified as a bicyclic, tricyclic, tetracyclic or polycyclic fused ring alkyl group, preferably a bicyclic or tricyclic, more preferably a 5-membered/5-membered or 5-membered/6-membered bicycloalkyl group. Non-limiting examples of "fused ring alkyl" 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 tetradecahydrophenanthryl.

"bridged cycloalkyl" means an aromatic system having 5 to 18 membered ring structures, containing two or more cyclic structures, sharing two all-carbon polycyclic groups with one another that are not directly attached to a carbon atom, one or more of the rings may contain one or more double bonds, but none of the rings has a fully conjugated pi electron, preferably a 6 to 12 membered bridged cycloalkyl, more preferably a 7 to 10 membered bridged cycloalkyl. Cycloalkyl groups which may be classified as bicyclic, tricyclic, tetracyclic or polycyclic bridged according to the number of constituent rings are preferably bicyclic, tricyclic or tetracyclic, more preferably bicyclic or tricyclic. Non-limiting examples of "bridged cycloalkyl" include, but are not limited to: (1 s,4 s) -bicyclo [2.2.1] heptyl, bicyclo [3.2.1] octyl, (1 s,5 s) -bicyclo [3.3.1] nonyl, bicyclo [2.2.2] octyl, and (1 r,5 r) -bicyclo [3.3.2] decyl.

"heterocyclyl", "heterocycle" or "heterocyclic" are used interchangeably herein to refer to a non-aromatic heterocyclic group in which one or more of the ring-forming atoms are heteroatoms, such as oxygen, nitrogen, sulfur atoms, and the like, including monocyclic, fused, bridged and spiro rings. Preferably having a 5 to 7 membered single 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-dioxothiomorpholinyl, piperidinyl, 2-oxopiperidinyl, pyrrolidinyl, 2-oxopyrrolidinyl, piperazin-2-one, 8-oxa-3-aza-bicyclo [3.2.1] octyl, and piperazinyl. The heterocyclic group may be substituted or unsubstituted.

"Spirocyclic radical"refers to a 5 to 18 membered, two or more cyclic structure, polycyclic groups having one atom in common with each other between the monocyclic rings, containing 1 or more double bonds within the rings, but no ring has a completely conjugated pi-electron aromatic system in which one or more of the ring atoms is selected from nitrogen, oxygen or S (O) _r (wherein r is selected from 0, 1 or 2) and the remaining ring atoms are carbon. Preferably a 6 to 14 membered spiroheterocyclyl group, more preferably a 7 to 10 membered spiroheterocyclyl group. The spirocycloalkyl group is classified into a single spiro heterocyclic group, a double spiro heterocyclic group or a multiple spiro heterocyclic group according to the number of common spiro atoms between rings, and preferably a single spiro heterocyclic group and a double spiro heterocyclic group. More preferably a 4-membered/4-membered, 4-membered/5-membered, 4-membered/6-membered, 5-membered/5-membered or 5-membered/6-membered single spiro heterocyclic 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 [3.5 ]]Nonyl and 5-oxaspiro [2.4 ]]A heptyl group.

"fused heterocyclyl" refers to an all-carbon polycyclic group containing two or more cyclic structures sharing a pair of atoms with each other, one or more of the rings may contain one or more double bonds, but none of the rings has a fully conjugated pi-electron aromatic system in which one or more of the ring atoms is selected from nitrogen, oxygen, or S (O) _r (wherein r is selected from 0, 1 or 2) and the remaining ring atoms are carbon. Preferably a 6 to 14 membered fused heterocyclic group, more preferably a 7 to 10 membered fused heterocyclic group. The number of constituent rings may be classified as a bicyclic, tricyclic, tetracyclic or polycyclic fused heterocyclic group, preferably a bicyclic or tricyclic, more preferably a 5-membered/5-membered or 5-membered/6-membered bicyclic fused heterocyclic group. 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]Dioxin (dioxin).

"bridged heterocyclyl" means a 5 to 14 membered, 5 to 18 membered, polycyclic group containing two or more cyclic structures sharing two atoms not directly attached to each other, one or more of the rings may contain one or more double bonds, but none of the rings has a fully conjugated pi electron aromatic system in which one or more of the ring atoms is selected from nitrogen, oxygen or S (O) _r (wherein r is selected from 0, 1 or 2) and the remaining ring atoms are carbon. Preferably a 6 to 14 membered bridged heterocyclyl, more preferably a 7 to 10 membered bridged heterocyclyl. Heterocyclic groups which may be classified as bicyclic, tricyclic, tetracyclic or polycyclic bridged according to the number of constituent rings are 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 [3.3.2]And (3) a decyl group.

"aryl" refers to a carbocyclic aromatic system containing one or two rings, wherein the rings may be linked together in a fused manner. The term "aryl" includes monocyclic or bicyclic aryl groups such as phenyl, naphthyl, tetrahydronaphthyl aromatic groups. Preferably aryl is C ₆ -C ₁₀ Aryl groups, more preferably aryl groups are phenyl and naphthyl. Aryl groups may 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. Preferred are bicyclic heteroaryl groups, examples of "heteroaryl" include, but are not limited to, the following: furyl, pyridyl, 2-oxo-1, 2-dihydropyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, 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, and the like,

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

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

"haloalkyl" refers to a group wherein the alkyl is optionally further substituted with one or more halogens, where alkyl is as defined herein.

"hydroxyalkyl" refers to a group in which the alkyl group is optionally further substituted with one or more hydroxyl groups, where 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 an-OH group.

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

"amino" means-NH ₂ 。

"cyano" refers to-CN.

"nitro" means-NO ₂ 。

"benzyl" means-CH ₂ -phenyl.

"carboxy" means-C (O) OH.

"carboxylate" refers to-C (O) O-alkyl or-C (O) O-cycloalkyl, wherein alkyl, cycloalkyl are as defined above.

"DMSO" refers to dimethyl sulfoxide.

"BOC" refers to t-butoxycarbonyl.

"Ts" refers to p-toluenesulfonyl.

"Ms" refers to sulfonyl.

"T3P" refers to propyl phosphoric anhydride.

"DPPA" refers to diphenyl azide phosphate.

"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, preferably up to 5, more preferably 1 to 3 hydrogen atoms in the group are independently substituted with a corresponding number of substituents. It goes without saying that substituents are only in their possible chemical positions, and that the person skilled in the art is able to determine (by experiment or theory) possible or impossible substitutions without undue effort. For example, amino or hydroxyl groups having free hydrogen may be unstable when bound to carbon atoms having unsaturated (e.g., olefinic) bonds.

"substituted" or "substituted" as used herein, unless otherwise indicated, means that the group may be substituted with one or more groups selected from the group consisting of: alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, alkenyl, hydroxy, nitro, cyano, cycloalkyl, heterocyclyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, 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 is ⁵ Selected from a hydrogen atom, an alkyl group, a cycloalkyl group, a heterocyclic group, an aryl group or a heteroaryl group, wherein the alkyl group, the cycloalkyl group, the heterocyclic group, the aryl group or the heteroaryl group is optionally further substituted with one or more groups selected from hydroxyl group, halogen, nitro group, cyano group, alkyl group, alkoxy group, haloalkyl group, haloalkoxy group, cycloalkyl group, heterocyclic group, aryl group, heteroaryl group, =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 by a substituent of (2);

R ⁶ and R is ⁷ Each independently of the otherIs selected from the group consisting of 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, the alkoxy group, the cycloalkyl group, the heterocyclic group, the aryl group, or the heteroaryl group is optionally further substituted with one or more groups selected from the group consisting of a hydroxyl group, a halogen, a nitro group, a cyano group, an alkyl group, an alkoxy group, a cycloalkyl group, a heterocyclic group, an aryl group, a heteroaryl group, =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 by a substituent of (2);

alternatively, R ⁶ And R is ⁷ Together with the atoms to which they are attached form a 4-8 membered heterocyclic group, wherein the 4-8 membered heterocyclic group contains one or more of N, O or S (O) _r And said 4-8 membered heterocyclyl is optionally further substituted with one or more substituents 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 Is substituted by a substituent of (2);

wherein R is ⁸ 、R ⁹ And R is ¹⁰ Each independently selected from the group consisting of hydrogen, alkyl, amino, cycloalkyl, heterocyclyl, aryl, or heteroaryl, wherein said alkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl is optionally further substituted with one or more substituents selected from the group consisting of hydroxy, halogen, nitro, amino, cyano, alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl, carboxyl, or carboxylate;

r is 0, 1 or 2.

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

Unless otherwise indicated, the structures described herein also include all stereoisomers (e.g., diastereomers, enantiomers and atropisomers and geometric (conformational) isomeric forms of such structures, e.g., the R and S configurations of each asymmetric center, (Z) and (E) double bond isomers, and (Z) and (E) conformational isomers.

"pharmaceutically acceptable salts" refers to certain salts of the above compounds which retain the original biological activity and are suitable for pharmaceutical use. The pharmaceutically acceptable salts of the compounds represented by formula (I) may be metal salts, amine salts with suitable acids.

"pharmaceutical composition" means a mixture comprising one or more of the compounds described herein or a physiologically acceptable salt or prodrug thereof, and other chemical components, and optionally other components such as physiologically acceptable carriers and excipients. The purpose of the pharmaceutical composition is to promote the administration to organisms, facilitate the absorption of active ingredients and thus exert biological activity.

Synthesis method of compound of the invention

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

the preparation method of the compound shown in the general formula (II) or the stereoisomer, the tautomer or the pharmaceutically acceptable salt thereof comprises the following steps:

reacting a compound of formula (IIA) with a compound of formula (IIB) under basic conditions, optionally further deprotecting to give a compound of formula (II);

wherein:

X ¹ as leaving group, preferably chloro, bromo, iodo, OMs or OTs; more preferably chlorine;

Ring B, R ¹ ～R ³ The definitions of X, Y, G, E and n are as described in formula (II).

Detailed Description

The invention will be further described with reference to the following examples, which are not intended to limit the scope of the invention.

Examples

The preparation of representative compounds represented by formula (I) and related structural identification data are presented in the examples. It must be noted that the following examples are given by way of illustration and not by way of limitation. ¹ HNMR spectra were determined using a Bruker instrument (400 MHz) and chemical shifts were expressed in ppm. Tetramethylsilane internal standard (0.00 ppm) was used. ¹ HNMR representation method: s=singlet, d=doublet, t=triplet, m=multiplet, br=broadened, dd=doublet of doublet, dt=doublet of triplet. If coupling constants are provided, they are in Hz.

The mass spectrum is measured by an LC/MS instrument, and the ionization mode can be ESI or APCI.

The thin layer chromatography silica gel plate uses a smoke table yellow sea HSGF254 or Qingdao GF254 silica gel plate, the specification of the silica gel plate used by the Thin Layer Chromatography (TLC) is 0.15 mm-0.2 mm, and the specification of the thin layer chromatography separation and purification product is 0.4 mm-0.5 mm.

Column chromatography generally uses tobacco stand yellow sea silica gel 200-300 mesh silica gel as a carrier.

In the following examples, unless otherwise indicated, all temperatures are in degrees celsius and unless otherwise indicated, various starting materials and reagents are either commercially available or synthesized according to known methods, all of which are used without further purification and unless otherwise indicated, commercially available manufacturers include, but are not limited to, shanghai Haohong biological medicine technologies, shanghai Shaoshao reagent, shanghai Pico medicine, saen chemical technologies (Shanghai) and Shanghai Ling Kai medicine technologies, and the like.

CD ₃ OD: deuterated methanol.

CDCl ₃ : deuterated chloroform.

DMSO-d ₆ : deuterated dimethyl sulfoxide.

In the examples, unless otherwise specified, the solution in the reaction means an aqueous solution.

Purifying the compound using an eluent system of column chromatography and thin layer chromatography, wherein the system is selected from the group consisting of: a: petroleum ether and ethyl acetate systems; b: methylene chloride and methanol systems; c: dichloromethane and ethyl acetate systems; d: dichloromethane and ethanol system; e: ethyl acetate and tetrahydrofuran systems; the volume ratio of the solvent is different according to the polarity of the compound, and can be adjusted by adding a small amount of acidic or alkaline reagent, such as acetic acid or triethylamine.

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 ] quinolin-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-carboxylic acid tert-butyl ester

7-bromo-4, 6-dichloro-8-fluoro-3-nitroquinoline 1a (550 mg,1.62mmol, prepared according to published patent WO 2019110751) and 3-aminoazetidine-1-carboxylic acid tert-butyl ester 1b (417.98 mg,2.43 mmol) were dissolved in acetonitrile (5 mL), cooled to 0deg.C, N-diisopropylethylamine (627.32 mg,4.85 mmol) was added dropwise, and the reaction was allowed to proceed to room temperature for a further 6 hours. The residue was concentrated under reduced pressure and purified by silica gel column chromatography (eluent: A system) to give tert-butyl 3- ((7-bromo-6-chloro-8-fluoro-3-nitroquinolin-4-yl) amino) azetidine-1-carboxylate 1c (400 mg, 840.87. Mu. Mol), 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-carboxylic acid tert-butyl ester

3- ((7-bromo-6-chloro-8-fluoro-3-nitroquinolin-4-yl) amino) azetidine-1-carboxylic acid 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), and heated to 90℃for reaction for 5 hours. After the reaction, the mixture was filtered while it was still 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 give crude 3- ((3-amino-7-bromo-6-chloro-8-fluoroquinolin-4-yl) amino) azetidine-1-carboxylic acid 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-carboxylic acid tert-butyl ester

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

MS m/z(ESI):455.8[M+1] ⁺

Fourth step

3- (8-chloro-6-fluoro-7- (2-fluoro-6-hydroxyphenyl) -1H- [1,2,3] triazolo [4,5-c ] quinolin-1-yl) azetidine-1-carboxylic acid 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 (350 mg, 766.37. Mu. Mol), (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. Mu. Mol) were dissolved in tetrahydrofuran (10 mL) and heated to 50℃under argon atmosphere and reacted overnight. After the completion of the reaction, the filtrate was filtered and concentrated under reduced pressure, and the obtained residue was separated and purified by silica gel column chromatography (eluent: A system) to give 1g (20 mg, 40.99. Mu. Mol) of tert-butyl 3- (8-chloro-6-fluoro-7- (2-fluoro-6-hydroxyphenyl) -1H- [1,2,3] triazolo [4,5-c ] quinolin-1-yl) azetidine-1-carboxylate, yield: 5.35%.

MS m/z(ESI):487.9[M+1] ⁺

Fifth step

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

1g (15 mg, 30.74. Mu. Mol) of tert-butyl 3- (8-chloro-6-fluoro-7- (2-fluoro-6-hydroxyphenyl) -1H- [1,2,3] triazolo [4,5-c ] quinolin-1-yl) azetidine-1-carboxylate (15 mg, 30.74. Mu. Mol) was dissolved in methylene chloride (5 mL), and a solution of HCl in 1, 4-dioxane (4M, 3 mL) was added and reacted at room temperature for 2 hours. After the reaction was completed, the mixture was concentrated under reduced pressure to give crude 2- (1- (azetidin-3-yl) -8-chloro-6-fluoro-1H- [1,2,3] triazolo [4,5-c ] quinolin-7-yl) -3-fluorophenol for 1H (11 mg, 28.37. Mu. Mol), yield: 92.27%.

MS m/z(ESI):388.0[M+1] ⁺

Sixth step

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

2- (1- (azetidin-3-yl) -8-chloro-6-fluoro-1H- [1,2,3] triazolo [4,5-c ] quinolin-7-yl) -3-fluorophenol 1H (15.9 mg, 41.00. Mu. Mol) was dissolved in dichloromethane (5 mL) for 1H, triethylamine (16.60 mg, 164.01. Mu. Mol) was added, cooled to 0℃and a solution of acryloyl chloride (7.42 mg, 82.01. Mu. Mol) in dichloromethane (2 mL) was added dropwise thereto, and the reaction was continued at 0℃for 1 hour. The reaction solution was concentrated under reduced pressure to give 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. Mu. Mol) which was directly used in the next reaction.

MS m/z(ESI):496.0[M+1] ⁺

Seventh step

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

2- (1- (1-Acylazetidin-3-yl) -8-chloro-6-fluoro-1H- [1,2,3]Triazolo [4,5-c ]]Quinoline-7-yl) -3-fluorophenyl acrylate 1i (20 mg, 40.33. Mu. Mol) was dissolved in a mixed solvent of tetrahydrofuran (5 mL) and water (2 mL), and lithium hydroxide monohydrate (8.46 mg, 201.67. Mu. Mol) was added thereto and reacted under an ice bath for 0.5 hours. Concentrated under reduced pressure to remove tetrahydrofuran, extracted with dichloromethane (60 mL. Times.2), the combined organic phases dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure, and the resulting residue was isolated and purified by preparative HPLC (separation column: boston Prime C18,150X 30mm I.D.,5 μm; mobile phase A: water (0.05% NH) ₃ H ₂ O+10mM NH ₄ HCO ₃ ) 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) azetidin-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 ] quinolin-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) azetidine-1-carboxylic acid 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 (354.89 mg,1.31 mmol), 4- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) naphthalen-2-ol 2a (300 mg, 656.89. Mu. Mol), potassium phosphate (418.30 mg,1.97 mmol) and X-PHOS Pd G2 (51.62 mg, 65.69. Mu. Mol) were dissolved in tetrahydrofuran (5 mL), protected with argon and heated to 50℃overnight. After the completion of the reaction, the residue was concentrated under reduced pressure and purified by silica gel column chromatography (eluent: A system) to give tert-butyl 3- (8-chloro-6-fluoro-7- (3-hydroxynaphthalen-1-yl) -1H- [1,2,3] triazolo [4,5-c ] quinolin-1-yl) azetidine-1-carboxylate 2b (340 mg, 653.90. Mu. Mol), 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) naphthalen-2-ol

3- (8-chloro-6-fluoro-7- (3-hydroxynaphthalen-1-yl) -1H- [1,2,3] triazolo [4,5-c ] quinolin-1-yl) azetidine-1-carboxylic acid tert-butyl ester 2b (100 mg, 192.32. Mu. Mol) was dissolved in methylene chloride (4 mL), and a 1, 4-dioxane solution (4M, 3 mL) of HCl was added to react at room temperature for 1 hour. After the reaction was completed, the mixture was concentrated under reduced pressure to give crude 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. Mu. Mol), 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 ] 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 ]]Quinolin-7-yl) naphthalen-2-ol 2c (80 mg, 190.55. Mu. Mol) was dissolved in dichloromethane (5 mL), cooled to 0℃and triethylamine (77.13 mg, 762.20. Mu. Mol) was added dropwise to a solution of acryloyl chloride (17.25 mg, 190.55. Mu. Mol) in dichloromethane (2 mL) and the reaction was continued at 0℃for 1 hour. After the reaction, the system was washed with saturated ammonium chloride solution (100 mL), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the resulting residue was separated and purified by preparative HPLC (separation column: boston Prime C18,150 ×30mm i.d.,5 μm; mobile phase a: water (0.05% nh) ₃ H ₂ O+10mM 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) azetidin-1-yl) prop-2-en-1-one

First step

(7-fluoro-4- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) benzo [ d ] thiazol-2-yl) carbamic acid tert-butyl ester

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

MS m/z(ESI):256.9[M+1-139] ⁺

¹ H NMR(400MHz,DMSO-d ₆ )δ1.32(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-carboxylic acid tert-butyl ester

Tert-butyl 3- (7-bromo-8-chloro-6-fluoro-1H- [1,2,3] triazolo [4,5-c ] quinolin-1-yl) azetidine-1-carboxylate 1e (512 mg,1.30 mmol), (7-fluoro-4- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) benzo [ d ] thiazol-2-yl) carbamate 3b (539.15 mg,1.18 mmol), potassium phosphate (501.17 mg,2.36 mmol), X-PHOS Pd G2 (185.53 mg, 236.11. Mu. Mol) was dissolved in tetrahydrofuran (10 mL), and heated to 50℃under argon atmosphere for 3 hours. After the completion of the reaction, the residue was concentrated under reduced pressure and purified by silica gel column chromatography (eluent: A system) to give tert-butyl 3c (270 mg, 419.20. Mu. Mol) of 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, 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-carboxylic acid tert-butyl ester 3c (270 mg, 419.20. Mu. Mol) was dissolved in dichloromethane (8 mL) and a solution of HCl in 1, 4-dioxane (4M, 8 mL) was added and reacted at room temperature for 1 hour. After the reaction was completed, the mixture was concentrated under reduced pressure to give crude 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. Mu. Mol) which was directly used in the next reaction.

MS m/z(ESI):443.8[M+1] ⁺

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) 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 ]]Quinolin-7-yl) -7-fluorobenzo [ d]The thiazol-2-amine 3d (186 mg, 419.05. Mu. Mol) was dissolved in dichloromethane (3 mL), triethylamine (127.21 mg,1.26 mmol) was added, cooled to 0deg.C, and a solution of acryloyl chloride (37.93 mg, 419.05. Mu. Mol) in dichloromethane (2 mL) was added dropwise and the reaction was continued at 0deg.C for 1 hour. After the reaction, the system was washed with saturated ammonium chloride solution (100 mL), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the resulting residue was separated and purified by preparative HPLC (separation column: boston Prime C18,150 ×30mm i.d.,5 μm; mobile phase a: water (0.05% nh) ₃ H ₂ O+10mM 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) azetidin-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. Mu. Mol) is chiral by SFCResolution (column number: chiralPak AD, 250X 30mm I.D.,10 μm; mobile phase: A for CO.) ₂ and B for Isopropanol (0.1% nh3h2 o); column pressure: 100bar; flow rate: 70mL/min; detection wavelength: 220nm; column temperature: after purification at 38 ℃) compound 3A and compound 3B belonging to one of the single configuration compound (shorter retention time) and the single configuration compound (longer retention time) respectively are obtained.

Single configuration compounds (shorter retention time):

MS m/z(ESI)：498.1[M+1] ⁺

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

Single configuration compounds (longer retention time):

136mg; retention time 3.908 min, 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 (0.05 g, 136.24. Mu. Mol, prepared according to published patent WO2016164675A 1) was dissolved in tetrahydrofuran (2 mL), cooled to-78℃and lithium diisopropylamide (1M, 299.73. Mu.L) was added dropwise, and the temperature was slowly raised to 25℃and the reaction was continued for 3 hours. LC-MS detection reaction was complete. To the reaction solution was added 5mL of water to quench the reaction, extracted with ethyl acetate (5 mL. Times.3), and the organic phases were combined, dried over anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure to give crude (7-bromo-4, 6-dichloro-8-fluoroquinolin-3-yl) methanol 4b (20 mg, 61.55. Mu. Mol) as a yellow oil in 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. Mu. Mol) was dissolved in methylene chloride (2 mL), and manganese dioxide (66.88 mg, 769.32. Mu. Mol) was added at 25℃and heated to 40℃to react for 16 hours. LC-MS detection reaction was complete. Filtering, and concentrating under reduced pressure to obtain 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-carboxylic acid tert-butyl ester

7-bromo-4, 6-dichloro-8-fluoroquinoline-3-carbaldehyde 4c (500 mg,1.55 mmol) and 3-hydrazinoazetidine-1-carboxylic acid tert-butyl ester 4d (434.84 mg,2.32mmol, prepared according to published patent WO2012004743A 1) were dissolved in ethanol (6 mL), triethylamine (470.0 0mg,4.64mmol) was added and reacted at 80℃for 3 hours. LC-MS detection reaction was complete. The residue was concentrated under reduced pressure and purified by column chromatography over silica gel (eluent: A system) to give tert-butyl 3- (7-bromo-8-chloro-6-fluoro-1H-pyrazolo [4,3-c ] quinolin-1-yl) azetidine-1-carboxylate 4e (0.2 g, 438.88. Mu. Mol) in 28.35% yield.

MS m/z(ESI):455.0[M+1] ⁺

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-carboxylic acid tert-butyl ester

Tert-butyl 3- (7-bromo-8-chloro-6-fluoro-1H-pyrazolo [4,3-c ] quinolin-1-yl) azetidine-1-carboxylate 4e (300 mg, 658.32. Mu. Mol), (2- ((tert-butoxycarbonyl) amino) -5, 7-difluorobenzo [ d ] thiazol-4-yl) boronic acid 4f (260.78 mg, 789.98. Mu. Mol, prepared according to published patent U.S. Pat. No. 20200115375A 1), potassium phosphate (279.47 mg,1.32 mmol) and X-PHOS Pd G2 (51.73 mg, 65.83. Mu. Mol) were dissolved in tetrahydrofuran (8 mL) and reacted overnight at 50 ℃. The reaction mixture was cooled to room temperature, filtered through celite, the filtrate was concentrated under reduced pressure, an appropriate amount of methanol was added to dissolve the residue, filtered again, and the filtrate was concentrated under reduced pressure, and the resulting residue was purified by column chromatography on silica gel (eluent: A system) to give 4g (21 mg, 31.77. Mu. Mol) of tert-butyl 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 in 4.83% yield.

MS m/z(ESI):661.2[M+1] ⁺

Fifth step

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

4g (21 mg, 31.77. Mu. Mol) of tert-butyl 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 (21 mg, 31.77. Mu. Mol) was dissolved in dichloromethane (3 mL) and HCl/1, 4-dioxane solution (4M, 3 mL) was added and reacted at room temperature for 1 hour. Concentrated under reduced pressure to give 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) in 79.78% yield.

MS m/z(ESI):463.0[M+3] ⁺

Sixth step

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 ]]The thiazol-2-amine was dissolved in dichloromethane (5 mL) for 4h (14.6 mg, 25.34. Mu. Mol), cooled to 0deg.C, triethylamine (8.56 mg, 84.62. Mu. Mol) and acryloyl chloride (2.55 mg, 28.21. Mu. Mol) were added and the reaction was continued at 0deg.C for 1 hour. After the completion of the reaction, the reaction mixture was washed with a saturated ammonium chloride solution (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure, and the obtained residue was separated and purified by preparative HPLC (separation column: boston Prime C18,150X 30mm I.D.,5 μm; mobile phase A: water (0.05% NH) ₃ H ₂ O+10mM NH ₄ HCO ₃ ) 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) azetidin-1-yl) prop-2-en-1-one 4 (1 mg, 1.84. Mu. Mol) in 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] triazolo [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-carboxylic acid 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-carboxylic acid 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. Mu. Mol) were dissolved in tetrahydrofuran (6 mL) and protected with argon and heated to 50℃overnight. After the reaction was completed, the filtrate was filtered and concentrated under reduced pressure, and the resulting residue was separated and purified by preparative HPLC (separation column: AKZONOBEL Kromasil; 250X 21.2mm I.D.;5 μm; mobile phase A:0.05% TFA+H2O; mobile phase B: acetonitrile; flow rate: 20 mL/min) to give the product, tert-butyl 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 5a (100 mg, 151.04. Mu. Mol), 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-carboxylic acid tert-butyl ester 5a (143 mg, 215.99. Mu. Mol) was dissolved in dichloromethane (5 mL), and a 1, 4-dioxane solution (4M, 3 mL) of hydrochloric acid was added to react at room temperature for 1 hour. Concentration under reduced pressure afforded the 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] triazolo [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 ] quinolin-7-yl) -5, 7-difluorobenzo [ d ] thiazol-2-amine 5b (99.75 mg, 215.98. Mu. Mol) was dissolved in dichloromethane (3 mL), cooled to 0℃and triethylamine (65.56 mg, 647.94. Mu. Mol) and acryloyl chloride (19.55 mg, 215.98. Mu. Mol) were added and the reaction was continued at 0℃for 1 hour. After the reaction, the reaction mixture was concentrated under reduced pressure, and the resulting residue was separated and purified by preparative HPLC (separation column: AKZONOBEL Kromasil; 250X 21.2mm I.D.;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. Mu. Mol), 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] triazolo [4,5-c ] quinolin-1-yl) azetidin-1-yl) prop-2-en-1-one 5 (30 mg, 58.15. Mu. Mol) was chiral resolved by SFC (column type: chiralCel OD, 250X 30mm I.D., 10. Mu.m; mobile phase: A for CO2 and B for Ethanol; column pressure: 100bar; flow rate: 80mL/min; detection wavelength: 220nm; column temperature: 38 ℃) to give, after purification, compound 5A and compound 5B, each belonging to one of the single configuration compound (shorter retention time) and the single configuration compound (longer retention time).

Single configuration compounds (shorter retention time):

MS m/z(ESI)：516.1[M+1] ⁺

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

Single configuration compounds (longer retention time):

10mg; retention time 8.542 min, 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) azetidin-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), a solution of benzoyl isothiocyanate 6b (23.13 g,141.70 mmol) in tetrahydrofuran (100 mL) was slowly added dropwise, and after the dropwise addition was completed, the reaction was continued at room temperature for 3 hours, LCMS monitored the starting materials all converted to intermediates, water (80 mL), sodium hydroxide (6.80 g,170.04 mmol) were added, and the reaction was heated to 80℃overnight. Cooled to room temperature, extracted with ethyl acetate (100 mL. Times.1), the organic phase was washed with saturated brine (100 mL. Times.1), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography (eluent: A system) to give 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 bromine (5.77 g,36.12 mmol) was slowly added dropwise, keeping the temperature below 30 ℃. After the completion of the dropwise addition, the reaction mixture was heated to 40℃and reacted overnight. The reaction solution was cooled, poured into water (500 mL), made alkaline with saturated sodium carbonate solution, extracted with ethyl acetate (300 mL. Times.1), the organic phase was washed with saturated brine (100 mL. Times.1), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the resulting residue was isolated and purified by silica gel column chromatography (eluent: A system) to give 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 0deg.C, boron tribromide (22.12 g,88.29mmol,8.51 mL) was added dropwise, and the mixture was allowed to react overnight at room temperature. The reaction solution was poured into ice water (300 mL), made alkaline with saturated sodium carbonate solution, extracted with ethyl acetate (500 mL. Times.1), and the organic phase was washed with saturated brine (100 mL. Times.1), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give the 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] ⁺

Fourth step

(4- ((tert-Butoxycarbonyl) oxy) -5-fluorobenzo [ d ] thiazol-2-yl) carbamic acid tert-butyl ester

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,108.58mmol,15.13 mL), 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 solution was separated, the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give tert-butyl (4- ((tert-butoxycarbonyl) oxy) -5-fluorobenzo [ d ] thiazol-2-yl) carbamate 6f (20 g,52.03 mmol), a crude product, yield: 95.83%.

MS m/z(ESI):385.1[M+1] ⁺

Fifth step

(5-fluoro-4-hydroxybenzo [ d ] thiazol-2-yl) carbamic acid tert-butyl ester

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

MS m/z(ESI):228.9[M+1-56] ⁺

Sixth step

2- ((Boc) amino) -5-fluorobenzo [ d ] thiazol-4-yl triflate

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

MS m/z(ESI):361.0[M+1-56] ⁺

Seventh step

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

2- ((Boc) amino) -5-fluorobenzo [ d ] thiazol-4-yl triflate 6h (5 g,12.01 mmol), pinacol biborate (24.40 g,96.07 mmol) were mixed in 1, 4-dioxane (80 mL), potassium acetate (3.54 g,36.03 mmol) was added, argon was displaced, stirred for 10 min, and [1,1' -bis (diphenylphosphino) ferrocene ] palladium dichloride (2.64 g,3.60 mmol) was added and stirred for 8 h at 100℃under argon. The pinacol biborate (24.40 g,96.07 mmol) and [1,1' -bis (diphenylphosphino) ferrocene ] palladium dichloride (2.64 g,3.60 mmol) were added and stirring was continued for 8 hours at 100℃under argon. The reaction solution was concentrated under reduced pressure, and the obtained residue was purified by silica gel column chromatography (eluent: A system) to give 6i (2 g,6.41 mmol) of (2- ((tert-butoxycarbonyl) amino) -5-fluorobenzo [ d ] thiazol-4-yl) boronic acid as a product in the yield: 53.36%.

MS m/z(ESI):312.9[M+1] ⁺

Eighth step

2- (((3-bromo-4-chloro-2-fluorophenyl) amino) methylene) -3-oxobutanoic acid ethyl ester

3-bromo-4-chloro-2-fluoroaniline 6j (3 g,13.37mmol, prepared according to published patent WO2016164675A 1), ethyl 2- (ethoxymethylene) -3-oxobutanoate 6k (3.73 g,20.05 mmol) were mixed and reacted overnight at 120 ℃. After the reaction, the reaction mixture was concentrated under reduced pressure, and the obtained residue was purified by silica gel column chromatography (eluent: A system) to give ethyl 2- (((3-bromo-4-chloro-2-fluorophenyl) amino) methylene) -3-oxobutyrate 6m (3 g,8.23 mmol), yield: 61.56%.

MS m/z(ESI):365.8[M+1] ⁺

Ninth step

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

Ethyl 2- (((3-bromo-4-chloro-2-fluorophenyl) amino) methylene) -3-oxobutanoate 6m (1 g,2.74 mmol) was dissolved in diphenyl ether (50 mL), and the temperature was raised to 250℃for 1.5 hours. After the reaction was completed, the reaction solution was cooled to room temperature, poured into petroleum ether (200 mL), the solid was precipitated, filtered, and the cake was rinsed with petroleum ether (50 mL), the solid was collected and dried to give 1- (7-bromo-6-chloro-8-fluoro-4-hydroxyquinolin-3-yl) ethan-1-one 6n (300 mg, 941.84. Mu. Mol), yield: 34.34%.

MS m/z(ESI):319.7[M+1] ⁺

Tenth step

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

1- (7-bromo-6-chloro-8-fluoro-4-hydroxyquinolin-3-yl) ethan-1-one 6n (300 mg, 941.84. Mu. Mol) was added to glacial acetic acid (5 mL), 3-hydrazinoazetidine-1-carboxylic acid tert-butyl ester 4d (264 mg,1.41 mmol) was added, and the mixture was heated to 80℃and reacted overnight. The reaction solution was concentrated under reduced pressure, ethyl acetate (20 mL) was added thereto, saturated sodium hydrogencarbonate solution was added dropwise to adjust the pH to basicity, extracted with ethyl acetate (10 mL. Times.3), washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, dried, concentrated under reduced pressure, and the obtained residue was separated and purified by silica gel column chromatography (eluent: A system) to give 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 (150 mg, 319.33. Mu. Mol), yield: 33.90%.

MS m/z(ESI):469.0[M+1] ⁺

Eleventh step

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-carboxylic acid tert-butyl ester

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 (140 mg, 298.04. Mu. Mol), (2- ((tert-butoxycarbonyl) amino) -5-fluorobenzo [ d ] thiazol-4-yl) boronic acid 6i (186.05 mg, 596.08. Mu. Mol), potassium phosphate (126.52 mg, 596.08. Mu. Mol) and X-PHOS Pd G2 (46.84 mg, 59.61. Mu. Mol) were dissolved in tetrahydrofuran (3 mL) and reacted overnight at 50 ℃. After the reaction was completed, the filtrate was filtered and concentrated under reduced pressure, and the resulting residue was isolated and purified by preparative HPLC (separation column: AKZONOBEL Kromasil; 250X 21.2mm I.D.;5 μm; mobile phase A:0.05% TFA+H2O; mobile phase B: acetonitrile; flow rate: 20 mL/min) to give tert-butyl 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 6q (50 mg, 76.09. Mu. Mol) in 25.53% yield.

MS m/z(ESI):656.8[M+1] ⁺

Twelfth step

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-pyrazolo [4,3-c ] quinolin-1-yl) azetidine-1-carboxylic acid tert-butyl ester 6q (50 mg, 76.09. Mu. Mol) was dissolved in dichloromethane (3 mL), and a 1, 4-dioxane solution (4M, 1 mL) of hydrochloric acid was added to react at room temperature for 1 hour. The reaction solution was concentrated under reduced pressure to give 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] ⁺

Thirteenth step

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

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 (33.05 mg, 72.33. Mu. Mol) was dissolved in dichloromethane (3 mL), cooled to 0℃and triethylamine (21.96 mg, 216.98. Mu. Mol) and acryloyl chloride (7.86 mg, 86.79. Mu. Mol) were added and the reaction was continued at 0℃for 1 hour. After the reaction, the reaction mixture was concentrated under reduced pressure, and the resulting residue was separated and purified by preparative HPLC (separation column: AKZONOBEL Kromasil; 250X 21.2mm I.D.;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. Mu. Mol), 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-acrylamidoglzetidin-3-yl) -8-chloro-6-fluoro-1H- [1,2,3] triazolo [4,5-c ] quinolin-7-yl) -2-amino-7-fluorobenzo [ b ] thiophene-3-carbonitrile

First step

(3-cyano-7-fluoro-4- (4, 5-tetramethyl-1, 3, 2-dioxaborane-2-yl) benzo [ b ] thiophen-2-yl) carbamic acid tert-butyl ester

(4-bromo-3-cyano-7-fluorobenzo [ b ]]Tert-butyl thiophen-2-yl) carbamate 7a (300 mg, 808.14. Mu. Mol, prepared by self-preparation according to WO 2021118877), 4', 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 (diphenylphosphinophenyl ether) palladium (II) dichloride (57.85 mg, 80.81. Mu. Mol) were added to 1, 4-dioxabicyclo (10 mL), protected with argon, and heated to 95℃for 4 hours. After the reaction was completed, the mixture was cooled to room temperature and concentrated under reduced pressure to prepare a liquid phase for separation (separation column AKZONOBEL Kromasil; 250X 21.2mm I.D.;5 μm,20mL/min; mobile phase A:0.05% TFA+H2O; mobile phase B: CH3 CN) to give (3-cyano-7-fluoro-4- (4, 5-tetramethyl-1, 3, 2-dioxaborane-2-yl) benzo [ B)]Thiophen-2-yl) ammoniaTert-butyl benzoate 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-carboxylic acid tert-butyl ester

Tert-butyl (3-cyano-7-fluoro-4- (4, 5-tetramethyl-1, 3, 2-dioxaborane-2-yl) benzo [ b ] thiophen-2-yl) carbamate 7b (109.91 mg, 262.76. Mu. Mol), 3- (7-bromo-8-chloro-6-fluoro-1H- [1,2,3] triazolo [4,5-c ] quinolin-1-yl) azetidine-1-carboxylate 1e (100 mg, 218.96. Mu. Mol), potassium phosphate (92.95 mg, 437.93. Mu. Mol), chloro (2-dicyclohexylphosphino-2 ',4',6 '-triisopropyl-1, 1' -biphenyl) [2- (2 '-amino-1, 1' -biphenyl) ] palladium (II) (17.21 mg, 21.90. Mu. Mol) were added to tetrahydrofuran (10 mL), and the mixture was heated to 50℃overnight under argon atmosphere. After the reaction, the reaction mixture was filtered through celite and concentrated under reduced pressure to give a liquid phase (separation column AKZONOBEL Kromasil; 250X 21.2mm I.D.;5 μm,20mL/min; mobile phase A:0.05% TFA+H2O; mobile phase B: CH3 CN) 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-carboxylic acid 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 ] quinolin-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-carboxylic acid tert-butyl ester 7c (50 mg, 74.84. Mu. Mol) was added to dichloromethane (3 mL), 4M dioxane hydrochloride solution (2 mL) was further added, and after completion of the reaction, the reaction was reacted at room temperature for 2 hours and concentrated under reduced pressure to give 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, the next reaction was directly carried out.

MS m/z(ESI):467.8[M+1] ⁺

Fourth step

4- (1- (1-acrylamidoglzetidin-3-yl) -8-chloro-6-fluoro-1H- [1,2,3] triazolo [4,5-c ] quinolin-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 ] quinolin-7-yl) -7-fluorobenzo [ b ] thiophene-3-carbonitrile 7d (35 mg, 74.81. Mu. Mol) was added to dichloromethane (5 mL), triethylamine (22.71 mg, 224.42. Mu. Mol) was added, and the mixture was cooled to 0℃and a dichloromethane solution of acryloyl chloride (6.77 mg, 74.81. Mu. Mol) was added dropwise thereto and reacted at room temperature for 2 hours. After the reaction was completed, the mixture was concentrated under reduced pressure to give a liquid phase (separation column AKZONOBEL Kromasil; 250X 21.2mm I.D.;5 μm,20mL/min; mobile phase A:0.05% TFA+H2O; mobile phase B: CH3 CN) to give 4- (1- (1-acrylamidoglitazone-3-yl) -8-chloro-6-fluoro-1H- [1,2,3] triazolo [4,5-c ] quinolin-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 ability of the inventive Compounds to covalently bind to KRAS G12C protein

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

The experimental procedure is briefly described as follows: recombinant human KRAS G12C protein (aa 1-169) was prepared at a concentration of 4uM for use using reaction buffer (20mM HEPES,150mM NaCl,1mM MgCl2,1mM DTT). Test compounds were dissolved in DMSO to prepare 10mM stock solution, which was then diluted with reaction buffer for use. First 1.5uL of test compound diluted with reaction buffer (final concentration of reaction system 3. Mu.M or 10. Mu.M) was added to the wells, followed by addition of 23.5uL of reaction buffer, mixing well, followed by addition of 25 uL of 4uM recombinant human KRAS G12C protein, incubation at room temperature for 5 or 15 minutes, termination of the reaction by addition of 5uL of acetic acid, and transfer of the sample to a sample bottle. Detection of the Exposure using the Agilent 1290/6530 instrument The ratio of covalent binding of the test compound to KRAS G12C protein, the sample was purified on a liquid chromatography column (XBridge Protein BEH C,

3.5 μm,2.1 mm. Times.50 mm), mobile phase A was 0.1% aqueous formic acid and mobile phase B was acetonitrile during the detection procedure, mobile phase elution procedure was: 0 to 0.5 minutes, maintaining the mobile phase A:95%,2.5 minutes, mobile phase a became 30% and held for 0.5 minutes, 3.1 minutes, mobile phase a became 95% and held for 1.9 minutes; flow rate: 0.5ml/min; finally, the data were analyzed using MassHunter Workstation Software Bioconfirm Version B.08.00 software to obtain the covalent Binding Rate (Binding Rate) of the test compound at a concentration of 3. Mu.M under incubation for 5min with KRAS G12C protein, as shown in Table 1.

TABLE 1 covalent binding Rate of the inventive Compounds with KRAS G12C protein

Conclusion the compounds of the present invention have better covalent binding rates with KRAS G12C protein.

Test example 2 inhibition of NCI-H358 cell proliferation assay by Compounds of the invention

The following method was used to determine the effect of the compounds of the invention on NCI-H358 cell proliferation. NCI-H358 cells (containing KRAS G12C mutation) were purchased from Shanghai institute of life sciences, china academy of sciences, and cultured in RPMI 1640 medium containing 10% fetal bovine serum, 100U penicillin, 100. Mu.g/mL streptomycin and 1mM Sodium Pyruvate. Cell viability by

Luminescent Cell Viability Assay kit (Promega, cat# G7573).

The experimental method is operated according to the steps of the instruction book of the kit,the following is a brief description: test compounds were prepared by first dissolving the test compounds in DMSO to prepare a 10mM stock solution, and then diluting the stock solution with medium to prepare test samples, wherein the final concentration of the compounds ranged from 1000nM to 0.015nM. Cells in the logarithmic growth phase were seeded at a density of 800 cells per well in 96-well cell culture plates at 37℃with 5% CO ₂ The culture was continued overnight in the incubator, followed by the addition of the test compound and continued for 120 hours. After the incubation was completed, a volume of 50. Mu.L of CellTiter-Glo assay solution was added to each well, and after shaking for 5 minutes, the mixture was allowed to stand for 10 minutes, followed by reading the Luminescence values of each well of the sample on a microplate reader using the Luminescence mode. The percent inhibition of compounds at each concentration point was calculated by comparison with the values of the control group (0.3% dmso), followed by nonlinear regression analysis of the compound concentration log-inhibition in GraphPad Prism 5 software to obtain IC compounds that inhibited cell proliferation ₅₀ The values are shown in Table 2.

TABLE 2 IC for inhibition of NCI-H358 cell proliferation by the compounds of the present invention ₅₀ Data

Conclusion the compounds of the present invention have a better 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 methods were used to determine the p-ERK1/2 inhibitory activity of the compounds of the invention on NCI-H358 cells. The method uses an Advanced phospho-ERK1/2 (Thr 202/tyr 204) kit (cat No. 64 AERPEH) from Cisbio, and the detailed experimental procedure is referred to the kit instructions. NCI-H358 cells (containing KRAS G12C mutation) were purchased from the China academy of sciences of Shanghai life sciences cell resource center.

The experimental procedure is briefly described as follows: NCI-H358 cells were cultured in RPMI 1640 complete medium containing 10% fetal bovine serum, 100U penicillin, 100 μg/mL streptomycin and 1mM Sodium Pyruvate. NCI-H358 cells were plated in 96-well plates 30000 per well, with medium being complete medium, at 37℃with 5% CO ₂ The cells were incubated overnight in an incubator. Test compounds were dissolved in DMSO to prepare a 10mM stock solution, then diluted with RPMI 1640 basal medium, and 90. Mu.L of RPMI 1640 basal medium containing the test compound at the corresponding concentration was added to each well, and the test compound was placed in a cell culture incubator for 3 hours and 40 minutes at a final concentration in the reaction system in the range of 1000nM to 0.015 nM. Subsequently, 10. Mu.L of hEGF (available from Roche under accession number 11376454001) in RPMI 1640 basal medium was added to a final concentration of 5nM and incubated in an incubator for 20 minutes. Cell supernatants were discarded, cells were washed with ice-bath PBS, after which 45. Mu.L of 1 Xcell phospho/total protein lysis buffer (Advanced phospho-ERK1/2 kit component) was added to each well for lysis, and 96-well plates were placed on ice for half an hour, followed by detection of lysates with reference to Advanced phospho-ERK1/2 (Thr 202/tyr 204) kit instructions. Finally, the fluorescence intensities of the wells at excitation wavelengths of 304nM, at which the emission wavelengths of 620nM and 665nM are measured on an microplate reader in TF-FRET mode, and the fluorescence intensity ratio of the wells 665/620 is calculated. The percent inhibition of the test compounds at each concentration was calculated by comparison with the fluorescence intensity ratio of the control group (0.1% dmso) and nonlinear regression analysis was performed by GraphPad Prism 5 software with the test compound concentration log-inhibition to obtain compound IC ₅₀ Values.

Conclusion the compounds of the invention have better proliferation inhibition effect on p-ERK1/2 in NCI-H358 cells.

Claims (16)

1. A compound of formula (I) or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof:

   

   wherein:

   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 with one or more R ^c Substituted;

   l is a bond or C ₁ -C ₆ An alkylene group; wherein said alkylene is optionally further substituted with one or more substituents selected from alkyl, halogen or hydroxy; preferably, L is a bond or-CH ₂ -、-CH ₂ CH ₂ -or-CH (CH) ₃ ) -; more preferably, L is a bond;

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

   Ring a is selected from the group consisting of:

   

   with the proviso 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 attached to ring A;

   when ring A is selected from

   

   When, -L-R ⁴ Absence of;

   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;

   R ^a selected from hydrogen atoms or fluorine;

   R ^b selected from hydrogen atoms, -CH ₂ F、-CHF ₂ 、

   

   R ^c The same OR different are each independently selected from hydrogen atom, alkyl, halogen, 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 R ⁵ Wherein said alkyl, cycloalkyl, heterocyclyl, aryl OR heteroaryl is optionally further substituted with one OR more substituents selected from alkyl, halo, 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 R ⁵ Is substituted by a substituent of (2);

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

   R ^f selected from a hydrogen atom, halogen, alkyl or alkoxy; wherein said alkyl or alkoxy is optionally further substituted with one or more substituents selected from halogen, hydroxy, cyano, alkyl or alkoxy; r is R ^f Preferably halogen, more preferably fluorine or chlorine;

   R ¹ selected from a hydrogen atom, halogen, alkyl or alkoxy; wherein said alkyl or alkoxy is optionally further substituted with one or more substituents selected from halogen, hydroxy, cyano, alkyl or alkoxy; r is R ¹ Preferably a hydrogen atom;

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

   R ^A each independently selected from alkyl, halo, 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 R ⁵ The method comprises the steps of carrying out a first treatment on the surface of the Wherein said alkyl, cycloalkyl, heterocyclyl, aryl OR heteroaryl is optionally further substituted with one OR more substituents selected from alkyl, halo, 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 R ⁵ Is substituted by a substituent of (2);

   with the proviso that when ring A is selected from

   

   When R is ² Not selected from

   

   R ³ Selected from a hydrogen atom, halogen, alkyl, alkoxy, cyano, haloalkyl or haloalkoxy, preferably a hydrogen atom;

   R ⁴ selected from a hydrogen atom, an aryl group or a heteroaryl group; wherein said aryl or heteroaryl is optionally further substituted with one or more R ^B Substitution; r is R ⁴ Heteroaryl groups are preferred.

   R ^B Each independently selected from alkyl, halo, 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 R ⁵ The method comprises the steps of carrying out a first treatment on the surface of the Wherein said alkyl, cycloalkyl, heterocyclyl, aryl OR heteroaryl is optionally further substituted with one OR more substituents selected from alkyl, halo, 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 R ⁵ Is substituted by a substituent of (2);

   R ⁵ selected from a hydrogen atom, an alkyl group, a cycloalkyl group, a heterocyclic group, an aryl group or a heteroaryl group, wherein the alkyl group, the cycloalkyl group, the heterocyclic group, the aryl group or the heteroaryl group is optionally further substituted with one or more groups selected from hydroxyl group, halogen, nitro group, cyano group, alkyl group, alkoxy group, haloalkyl group, haloalkoxy group, cycloalkyl group, heterocyclic group, aryl group, heteroaryl group, =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 by a substituent of (2);

   R ⁶ and R is ⁷ Each independently selected from a hydrogen atom, hydroxy, halogen, alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl or heteroaryl, wherein said alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl or heteroaryl is optionally further substituted with one or more substituents 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 Is substituted by a substituent of (2);

   alternatively, R ⁶ And R is ⁷ Together with the atoms to which they are attached form a 4-8 membered heterocyclic group, wherein the 4-8 membered heterocyclic group contains one or more of N, O or S (O) _r And said 4-8 membered heterocyclic group optionally furtherIs substituted with 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 Is substituted by a substituent of (2);

   R ⁸ 、R ⁹ and R is ¹⁰ Each independently selected from the group consisting of hydrogen, alkyl, amino, cycloalkyl, heterocyclyl, aryl, or heteroaryl, wherein said alkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl is optionally further substituted with one or more substituents selected from the group consisting of hydroxy, halogen, nitro, amino, cyano, alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl, carboxyl, or carboxylate;

   R ¹¹ Selected from a hydrogen atom, halogen, alkyl, alkoxy, cyano, nitro, amino, hydroxy, haloalkyl, haloalkoxy, or = O;

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

   R ¹³ Selected from alkyl groups, preferably methyl;

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

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

   

   wherein:

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

   -G-E is selected from:

   

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

   R ² selected from phenyl, naphthyl, pyridinyl, benzothiazolyl, or benzopyrazolyl, wherein said phenyl, naphthyl, pyridinyl, benzothiazolyl, or benzopyrazolyl is optionally further substituted with one or more R ^A Substitution;

   R ^A each independently selected from halogen, hydroxy, alkyl, alkoxy, cycloalkyl or-NR ⁶ R ⁷ Wherein said alkyl or alkoxy is optionally further substituted with one or more groups selected from halogen or-NR ⁶ R ⁷ Is substituted by a substituent of (2); wherein said halogen is preferably fluorine;

   R ⁶ and R is ⁷ Each independently selected from a hydrogen atom or an alkyl group, wherein the alkyl group is preferably methyl.
8. A compound according to any one of claims 1-7, or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, wherein R ² Selected from:

   

   
9. a compound according to any one of claims 1-8, or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, wherein R ³ Selected from hydrogen atomsAnd (5) a seed.
10. The 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. a compound according to any one of claims 1-10, or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, wherein the compound is:

    
12. a process for the preparation of a compound of general formula (II) according to any one of claims 2 to 11, or a stereoisomer, tautomer or pharmaceutically acceptable salt thereof, which process comprises the steps of:

    

    reacting a compound of formula (IIA) with a compound of formula (IIB) under basic conditions, optionally further deprotecting to give a compound of formula (II);

    wherein:

    X ¹ preferably selected from chlorine, bromine, iodine, OMs or OTs, more preferably chlorine;

    ring B, R ¹ ～R ³ The definitions of X, Y, G, E and n are as defined in claim 2.
13. A pharmaceutical composition comprising an effective amount of a compound according to any one of claims 1 to 11, or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, and optionally a pharmaceutically acceptable carrier, excipient, or combination thereof.
14. Use of a compound according to any one of claims 1 to 11, or a stereoisomer, tautomer or pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 13, for the preparation of a K-Ras gtpase inhibitor, wherein the K-Ras gtpase inhibitor is preferably a KRAS G12C inhibitor.
15. Use of a compound according to any one of claims 1 to 11, or a stereoisomer, tautomer or pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 13, for the manufacture of a medicament for the treatment of a disease mediated by KRAS mutations, wherein the disease mediated by KRAS mutations is preferably selected from cancer, wherein the cancer is preferably selected from pancreatic cancer, colorectal cancer, lung cancer, multiple myeloma, uterine cancer, cholangiocarcinoma, gastric cancer, bladder cancer, diffuse large B-cell lymphoma, rhabdomyosarcoma, cutaneous squamous cell carcinoma, cervical cancer, testicular germ cell carcinoma, particularly preferably the cancer is pancreatic cancer, colorectal cancer and lung cancer, wherein the KRAS mutations are preferably KRAS G12C mutations.
16. Use of a compound according to any one of claims 1 to 11, or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 13, for the manufacture of a medicament for the treatment of cancer, wherein the cancer is preferably selected from pancreatic cancer, colorectal cancer, lung cancer, multiple myeloma, uterine cancer, cholangiocarcinoma, gastric cancer, bladder cancer, diffuse large B-cell lymphoma, rhabdomyosarcoma, cutaneous squamous cell carcinoma, cervical cancer, testicular germ cell carcinoma, more preferably from pancreatic cancer, colorectal cancer and lung cancer; wherein the lung cancer is preferably non-small cell lung cancer.