CN115322158A - Substituted quinazolines as KRASG12C protein inhibitors - Google Patents

Abstract

The invention discloses a substituted quinazoline compound as a KRAS ^G12C protein inhibitor, which belongs to the field of biomedicine. The present invention provides a substituted quinazoline compound having a structure represented by general formula (I). The substituted quinazoline compound has good KRAS ^G12C protein inhibitory activity, and has good KRAS ^G12C inhibitory activity and metabolic stability while ensuring good KRAS G12C inhibitory activity. In the case of sex, it also has better target inhibition selectivity.

Description

Substituted quinazolines as KRASG12C protein inhibitors

technical field

The invention relates to a substituted quinazoline compound as a KRAS ^G12C protein inhibitor, belonging to the field of biomedicine.

Background technique

The KRAS gene (Kirsten Rat Sarcoma Viral Oncogene Homolog, Kirsten Rat Sarcoma Viral Oncogene Homolog) mutation was first discovered in NSCLC genes in 1984. The KRAS gene encodes a GTP/GDP binding protein belonging to the small GTPase family, KRAS The protein is composed of six β-sheets and five α-helices. The KRAS protein is 20kDa in size. The function of the KRAS protein is as a molecular switch to turn on/off the transduction of tyrosine kinases and related cell signaling pathways. The molecules involved in signal transduction in the cell signaling pathway in which KRAS protein participates include EGFR, Raf, MEK, MAPK, PI3K, and Akt. The activation of KRAS is achieved after linking with GTP under the action of guanine nucleotide exchange factors (including SOS protein, RAF, PI3K, etc.), and the inactivation of KRAS protein is through linking with GDP. When GTP is connected to KRAS protein, the conformation of KRAS changes, and the changed conformation prompts KRAS to interact with downstream signaling regulatory proteins such as Raf, PI3K, Ral-GDS, etc., leading to the proliferation and growth of cancer cells. The inactivation of KRAS is through the interaction between KRAS and GAPs (GTPase activating protein), thereby enhancing the function of the GTPase of KRAS protein and realizing the connection between KRAS and GDP. Through this mechanism, KRAS plays a role in cell proliferation and regulation of cell signaling. transduction plays an important role.

The KRAS protein consists of three structural domains with significant functional differences and a total of 188 amino acids. The three structural domains are the effector region, the allosteric region, and the hypervariable region. The effector region is the KRAS protein substrate binding unit, which is mainly combined with effector proteins and nucleotides, and is divided into three regions: loop region, switch1 region, and switch2 region. The amino acid sequence relative to the effector region is highly conserved (100% homology in different RAS subtypes), and the hypervariable region and allosteric region have 86% and 15% amino acid sequence identity compared with other RAS subtypes, respectively. The allosteric region is related to the interaction and dimerization of KRAS with the membrane, while the hypervariable region is related to the recognition of KRAS and the cell membrane.

KRAS mutations are associated with 22% of human cancers, including lung cancer (17%), colon cancer 33%, and pancreatic cancer 61%. 80% of KRAS mutations occur at codon 12, causing single amino acid substitutions, glycine to cysteine (G12C, 14%), aspartic acid (G12D, 36%), and valine (G12V, 23%), and the RAS mutation in lung cancer is mainly KRAS mutation (44%).

Due to the high expression of KRAS ^G12C mutation in tumor patients, it will also make patients resistant to other targeted drugs, KRAS ^G12C protein has attracted more and more experts and scholars' attention. However, after nearly three decades of hard work, it is still full of difficulties to directly develop small molecule inhibitors for clinical use against the KRAS ^G12C target. ) and use it to treat various diseases, such as cancer.

Alexis has obtained the first KRAS ^G12C inhibitor, ARS-1620, targeting the allosteric site of KRAS ^G12C , and many types of KRAS ^G12C inhibitors have been obtained based on the core structure of ARS-1620. Currently, the only One approved KRAS ^G12C inhibitor is Sotorasib (AMG-510) from Amgen, and another KRAS ^G12C inhibitor that is being applied for marketing is Adagrasib (MRTX849).

Contents of the invention

In order to solve the above problems in the prior art, the present invention provides a compound with general formula (I) (including its stereoisomers and tautomers) or a pharmaceutically acceptable salt thereof, a pharmaceutical composition and use. The compound of the invention has the characteristic of targeting and inhibiting KRAS ^G12C protein.

The present invention aims to provide a class of substituted quinazoline compounds with a general structural formula as shown in Formula 1, or their respective optical isomers, crystal forms, pharmaceutically acceptable salts, hydrates or solvates:

in:

m is 1 or 2;

R is selected from methoxy, hydroxyl, amino, halogen (F, Cl, Br, I), methylamino, ^C1 -C2 alkyl, C1-C2 alkoxy;

Y is an oxygen atom or a nitrogen atom;

R ² is selected from aryl or heteroaryl, and the aryl or heteroaryl can be substituted by 1-2 of the following groups: halogen, C1-C2 alkoxy, hydroxyl, amino, cyano, C1- C2 alkyl, C3-C6 cycloalkyl;

为

R ³ is monosubstituted to trisubstituted, selected from hydrogen, methyl, hydroxymethyl, or

for

R ⁴ is independently selected from hydrogen, fluorine, and methoxy;

为

R ⁵ is monosubstituted to trisubstituted, selected from hydrogen, methyl, hydroxymethyl; or

for

In one embodiment of the invention, R ¹ is selected from methoxy or methylamino.

^In one embodiment of the present invention, R is selected from aryl or heteroaryl, and the aryl or heteroaryl can be substituted by one of the following groups: halogen (fluorine, chlorine, bromine, iodine), C1-2 alkoxy, cyano.

优选为

其中R^3’和R^3”分别独立选自氢、甲基、羟甲基；或者

为

In one embodiment of the invention,

preferably

Wherein R ^3' and R ^3" are independently selected from hydrogen, methyl, hydroxymethyl; or

for

In one embodiment of the present invention, R ^3' and R ^3" are not hydrogen at the same time.

^In one embodiment of the invention, R4 is hydrogen or fluorine.

中R⁵优选为甲基、羟甲基，或者

为

In one embodiment of the invention,

In R ⁵ is preferably methyl, hydroxymethyl, or

for

In one embodiment of the present invention, Y is oxygen or nitrogen.

In one embodiment of the present invention, aryl refers to a substituted or unsubstituted benzene ring, naphthalene ring.

In one embodiment of the invention, heteroaryl refers to a substituted or unsubstituted heteroaryl which is a mono- or bicyclic aromatic ring system having 5 to 12 ring atoms, wherein at least one atom in the ring system is Heteroatoms selected from N, O and S. This definition also includes ring systems in which heteroaryl is fused to a benzene ring. Examples of suitable "heteroaryl rings" or "heteroaryl groups" are furyl, furanyl, imidazolyl, 1H-indazolyl, indolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, pyrimidinyl, pyrazinyl, pyrazolyl, pyridyl, pyrrolyl, Thiazolyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,3,4-thiadiazolyl and thienyl, Benzimidazolyl, benzofuryl, benzothienyl, benzoxazolyl, benzothiazolyl, benzotriazolyl, benzotetrazolyl, benzisoxazolyl, benzisothiazolyl , Quinolinyl.

Preferred:

In some embodiments of the present invention, m is selected as 1.

In certain embodiments of the invention, Y is oxygen.

^In certain embodiments of the invention, R4 is independently selected from hydrogen.

In certain embodiments of the invention, R ¹ is selected from methoxy.

In some embodiments of the present invention, R ² is selected from halo(chlorofluoro)aryl, alkoxy(C1-C2)aryl, and the inventors have found that such groups have more favorable hydrophobic interactions resulting in better protein inhibitory activity.

In certain embodiments of the invention, R ³ is selected from methyl.

^In certain embodiments of the invention, R is selected from hydrogen.

In some embodiments of the present invention, the compound has the following structure and stereo configuration:

In some embodiments of the present invention, in order to obtain better KRAS ^G12C protein inhibitory activity, preferably, the compound has the following structure and stereo configuration:

In one embodiment of the present invention, the pharmaceutically acceptable salt is an inorganic salt or an organic salt, and the inorganic salt includes hydrochloride, hydrobromide, hydroiodide, sulfate, bisulfate, Nitrates, phosphates, acid phosphates; said organic salts are selected from the group consisting of acetates, trifluoroacetates, propionates, pyruvates, glycolates, oxalates, malonates, Fumarate, Maleate, Lactate, Malate, Citrate, Tartrate, Methanesulfonate, Isosulfonate, Benzenesulfonate, Salicylate.

The present invention also provides the above-mentioned substituted quinazoline compounds, or their optical isomers, crystal forms, pharmaceutically acceptable salts, hydrates or solvates used in the preparation of prevention or treatment of KRAS ^G12C protein-mediated use in medicines for diseases.

The present invention also provides a pharmaceutical composition comprising the above-mentioned substituted quinazoline compound, or its optical isomers, crystal forms, pharmaceutically acceptable salts, hydrates or solvates.

In one embodiment of the present invention, the pharmaceutical composition further includes: a pharmaceutically acceptable carrier, excipient or diluent.

The present invention also provides the use of the above-mentioned substituted quinazoline compounds, or their optical isomers, crystal forms, pharmaceutically acceptable salts, hydrates or solvates in preparing KRAS ^G12C mutation inhibitors.

The present invention also provides a method for treating cancer, which comprises administering a therapeutically effective amount of the aforementioned compound, its pharmaceutically acceptable salt, stereoisomer, solvate or prodrug to a subject in need thereof, Or any combination of the above, or the step of administering the above pharmaceutical composition.

The present invention also provides a medicine for treating cancer, including the above-mentioned substituted quinazoline compounds, or their optical isomers, crystal forms, pharmaceutically acceptable salts, hydrates or solvates.

In one embodiment of the invention, the cancer is pancreatic ductal carcinoma, colorectal cancer, multiple myeloma, lung cancer, cutaneous melanoma, endometrioid carcinoma, uterine carcinosarcoma, thyroid cancer, acute myeloid Leukemia, urothelial carcinoma of the bladder, gastric cancer, cervical cancer, squamous cell carcinoma of the head and neck, diffuse large B-cell lymphoma, esophageal cancer, chronic lymphocytic leukemia, squamous cell carcinoma of the lung, small cell lung cancer, renal papillary Cell carcinoma, adenoid cystic carcinoma, chromophobe renal cell carcinoma, liver carcinoma, invasive carcinoma of the breast, squamous cell carcinoma of the cervix, serous adenocarcinoma of the ovary, adrenocortical carcinoma, prostate cancer, neuroblastoma, low-grade glia of the brain tumor, glioblastoma, medulloblastoma, squamous cell carcinoma of the esophagus, clear cell carcinoma of the kidney, osteosarcoma, small cell carcinoma of the ovary, rhabdoid tumor, sarcoma, neuroendocrine tumor of the small intestine, T-cell prolymphocytic leukemia .

Beneficial effect:

The compound with the structure represented by the general formula (I) provided by the present invention has a certain selectivity and a relatively novel structure under the condition of ensuring good KRAS ^G12C inhibitory activity and metabolic stability.

Detailed ways

The technical solutions of the present invention will be described in detail below in conjunction with embodiments.

"(1-2C)alkyl" in the present invention means a saturated branched or branched monovalent hydrocarbon group of 1 to 2 carbon atoms, respectively. Examples include, but are not limited to methyl, ethyl. "(1-3C)alkyl" can be defined by analogy, specific examples include but not limited to methyl, ethyl, 1-propyl, 2-propyl.

"Halogen" in the present invention means fluorine, chlorine, bromine and iodine.

"Pharmaceutically acceptable salts" in the present invention means those salts which retain the biological effectiveness and properties of the parent compound. The term "salt" refers to any salt of a compound according to the invention prepared from inorganic or organic acids or bases and internally formed salts. Usually, such salts have a physiologically acceptable anion or cation.

"Administering" or "administering" a compound to an individual in the present invention means providing a compound of the invention to an individual in need of treatment.

References throughout this specification to "an embodiment" or "an embodiment" or "in another embodiment" or "in some embodiments" or "in some embodiments of the present application" mean that at least An embodiment includes specific reference elements, structures or features described in relation to that embodiment. Thus, the phrases "in one embodiment" or "in an embodiment" or "in another embodiment" or "in certain embodiments" or "in some embodiments" or "in some "in an embodiment" does not necessarily all refer to the same embodiment. Furthermore, particular elements, structures or characteristics may be combined in any suitable manner in one or more embodiments.

<compound or pharmaceutically acceptable salt>

Further, the compounds of formula (I) or their salts may be isolated in the form of solvates, and any such solvates therefore fall within the scope of the present invention.

Compounds of general formula (I) also include compounds which differ only in the presence of one or more isotopically enriched atoms. For example, compounds of the present invention include within the scope of the present invention compounds wherein one or more hydrogen atoms are replaced by deuterium or tritium, or one or more carbon atoms are replaced by a 13C- or 14C-enriched carbon.

The compounds of the present invention or pharmaceutically acceptable salts thereof can be formulated as solid preparations for oral administration, including, but not limited to, capsules, tablets, pills, powders, granules and the like. In these solid dosage forms, the compound of general formula (I) of the present invention is mixed as an active ingredient with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate. Or mixed with the following ingredients: (1) fillers or solubilizers, such as starch, lactose, sucrose, glucose, mannitol and silicic acid, etc.; (2) binders, such as hydroxymethylcellulose, alginate, gelatin , polyvinylpyrrolidone, sucrose, gum arabic, etc.; (3) humectants, such as glycerin, etc.; (4) disintegrants, such as agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain silicic acid salt and sodium carbonate, etc.; (5) slow solvents, such as paraffin, etc.; (6) absorption accelerators, such as quaternary ammonium compounds, etc.; (7) wetting agents, such as cetyl alcohol and glyceryl monostearate, etc.; (8) ) Adsorbents, such as kaolin, etc.; (9) Lubricants, such as talc, calcium stearate, solid polyethylene glycol, sodium lauryl sulfate, etc., or mixtures thereof. Capsules, tablets, pills may also contain buffering agents.

The solid dosage forms such as tablets, dragees, capsules, pills, and granules can be crystal-coated or microencapsulated with coatings and shell materials, such as enteric coatings and others well known in the art. They may contain opacifying agents, and the release of the active ingredient in such compositions may be in a delayed manner in a certain part of the alimentary canal. Examples of usable embedding components are polymeric substances and waxy substances. If necessary, the active ingredient can also be in the form of microcapsules with one or more of the above-mentioned excipients.

The compounds of the present invention or pharmaceutically acceptable salts thereof can be formulated into liquid dosage forms for oral administration, including, but not limited to, pharmaceutically acceptable emulsions, solutions, suspensions, syrups, tinctures and the like. In addition to the compound of general formula (I) or a pharmaceutically acceptable salt thereof as an active ingredient, the liquid dosage form may contain inert diluents conventionally used in the art, such as water and other solvents, solubilizers and emulsifiers, e.g., ethanol , isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butanediol, dimethylformamide and oils, especially cottonseed oil, peanut oil, corn oil, olive oil, castor oil, sesame oil, etc. or mixtures of these substances, etc. Besides these inert diluents, the liquid dosage forms of the present invention may also include conventional adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring and perfuming agents, and the like.

Such suspending agents include, for example, ethoxylated stearyl alcohol, polyoxyethylene sorbitol, and sorbitan, microcrystalline cellulose, agar-agar and the like or mixtures of these substances.

The compounds of the present invention and their pharmaceutically acceptable salts can be formulated into dosage forms for parenteral injection, including, but not limited to, physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions, and Sterile powder for reconstitution into sterile injectable solutions and dispersions. Suitable carriers, diluents, solvents, excipients include water, ethanol, polyols and suitable mixtures thereof.

The compounds of the present invention or pharmaceutically acceptable salts thereof can be formulated into dosage forms for topical administration, including ointments, powders, suppositories, drops, sprays, inhalants and the like. The compound of general formula (I) of the present invention or a pharmaceutically acceptable salt thereof as an active ingredient under sterile conditions and a physiologically acceptable carrier and optional preservatives, buffers, and propellants that may be required if necessary Mix together.

The pharmaceutical composition of the present invention includes the compound of general formula (I) or a pharmaceutically acceptable salt thereof as an active ingredient, as well as pharmaceutically acceptable carriers, excipients and diluents. When preparing a pharmaceutical composition, usually the compound of the general formula (I) of the present invention or a pharmaceutically acceptable salt thereof is mixed with a pharmaceutically acceptable carrier, excipient or diluent, and the unit dose of its preparation formula contains 0.05 g-210 mg of the compound of formula (I) or a pharmaceutically acceptable salt thereof, preferably, the unit dose of the formulation contains 0.1 mg-100 mg of the compound of formula (I).

<purpose>

The present invention also provides the effect of the compound of general formula (I) or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating KRAS ^G12C protein-related cancer.

The compound of general formula (I) or its pharmaceutically acceptable salt closes the signaling pathway that promotes the proliferation of cancer cells by maintaining the KRAS ^G12C inactivation state, and is used for the treatment of cancers in mammals including humans. The aforementioned mammals, including humans, are administered a therapeutically effective amount of the compound of the above formula (I) or a pharmaceutically acceptable salt thereof.

A "therapeutically effective amount" is an amount of a compound of the invention effective to produce a biological or medical response (eg, decrease or inhibit enzymatic protein activity, or ameliorate symptoms, alleviate disease conditions, slow or delay disease progression, or prevent disease) in an individual.

The cancers mentioned in the present invention include pancreatic ductal carcinoma, colorectal cancer, multiple myeloma, lung cancer, skin melanoma, endometrial carcinoma, uterine carcinosarcoma, thyroid cancer, acute myelogenous leukemia, bladder and urinary tract Skin cancer, gastric cancer, cervical cancer, head and neck squamous cell carcinoma, diffuse large B-cell lymphoma, esophageal cancer, chronic lymphocytic leukemia, lung squamous cell carcinoma, small cell lung cancer, renal papillary cell carcinoma, adenoid Cystic carcinoma, chromophobe renal cell carcinoma, liver cancer, breast invasive carcinoma, cervical squamous cell carcinoma, ovarian serous adenocarcinoma, adrenocortical carcinoma, prostate cancer, neuroblastoma, low-grade glioma, glioblastoma Cytoma, medulloblastoma, squamous cell carcinoma of the esophagus, clear cell carcinoma of the kidney, osteosarcoma, small cell carcinoma of the ovary, rhabdoid tumor, sarcoma, neuroendocrine tumor of the small intestine, T-cell prolymphocytic leukemia.

The compounds of the present invention or pharmaceutically acceptable salts thereof can be administered to mammals including humans, and can be administered orally, rectally, parenterally (intravenously, intramuscularly or subcutaneously), topically (powder, ointment, drops) ) or intratumoral administration.

The compounds of the present invention and their pharmaceutically acceptable salts, as well as pharmaceutical compositions containing such compounds or their salts, can be used in combination with other antineoplastic drugs or therapies such as radiotherapy or chemotherapy.

The following examples are used to illustrate but not limit the synthesis method of the compound of general formula (I).

Temperatures are in degrees Celsius. If not stated otherwise, reagents were purchased from commercial suppliers and used without further purification. The structures of final products, intermediates and starting materials are confirmed by standard analytical methods, eg, spectral characterization, MS, NMR, and abbreviations used are those conventional in the art.

Preparation of Intermediate A:

The preparation method of 4-chloro-2-methoxyquinazoline-7-acetate (intermediate A):

The preparation of step a, 4-hydroxyl-2-nitrobenzoic acid:

4-Methoxy-2-nitrobenzoic acid (20 g, 101.5 mmol) was suspended in 100 mL of 40% hydrobromic acid and 100 mL of acetic acid. React at 120° C. for 20 h, and TLC detects that the reaction is complete. The reaction system was cooled to room temperature, concentrated under reduced pressure to obtain the crude product; the crude product was dissolved in 150 mL of dichloromethane, saturated aqueous sodium bicarbonate was added, the water phase was separated, the pH was adjusted to about 2 with 1M hydrochloric acid, extracted with ethyl acetate, and the combined The extract was washed with saturated brine, dried over anhydrous sodium sulfate, concentrated and dried to obtain 20.2 g of crude 2-amino-5-hydroxy 4-methoxybenzoic acid as a yellow solid.

The preparation of step b, 2-amino-4-hydroxybenzoic acid:

In a hydrogen atmosphere at room temperature, 100 mL of a mixed solvent of 2-amino-5-hydroxy 4-methoxybenzoic acid (20.2 g, 110.38 mmol), 2 g of Pd-C (10%) and methanol-tetrahydrofuran (V ₁ : V ₂ =1:1) the mixture was stirred for 3 h, and the reaction was complete as detected by TLC. After filtering, washing with methanol, the methanol extracts were combined, and the solvent was distilled off under reduced pressure to obtain a black solid product (10.47 g, yield 62%).

Step c, 2-methoxyquinazoline-4, the preparation of 7-diol:

To a mixture of 2-amino-5-hydroxy 4-methoxybenzoic acid (10.47 g, 68.43 mmol) and 2-methoxyethanol (150 mL) was added oxymethylisourea hydrochloride (15.12 g, 136.86 mmol) , heated to 125° C. to react overnight, and TLC detected that the reaction was complete. After the reaction solution was cooled, it was distilled under reduced pressure, and the residue was poured into 250 mL of water, and the pH value was adjusted to 7 by adding ammonia water dropwise, filtered, the filter cake was washed with water, and the filter cake was vacuum-dried to obtain a brown solid 2-methoxyquinazoline-4 , Synthesis of 7-diol (8.93g, yield 68%) step d, 4-hydroxyl-2-methoxyquinazolin-7-yl acetate:

Add pyridine (7.36g, 93.06mmol) to a mixture of 2-methoxyquinazoline-4,7-diol (8.93g, 46.53mmol) and 65mL of acetic anhydride, heat to 80°C for 1h, and detect the reaction by TLC completely. After the reaction solution was cooled and distilled under reduced pressure, the residue was poured into 100 mL of water, filtered, and the filter cake was vacuum-dried to obtain a gray solid 4-hydroxy-2-methoxyquinazolin-7-yl acetate (10.57 g, produced rate 97%).

The preparation of step e, 4-chloro-2-methoxyquinazoline-7-acetate:

To the mixture of 4-hydroxy-2-methoxyquinazoline-7-acetate (10.57g, 45.13mmol) and 1,2-dichloroethane (80mL), drop 80mL of thionyl chloride, then Add 2 drops of N,N'dimethylformamide dropwise, after the drop is complete, reflux at 80°C, and TLC detects that the reaction is complete. The solvent was evaporated under reduced pressure, the residue was poured into 150 mL of water, and saturated sodium bicarbonate aqueous solution was slowly added dropwise to the residue under an ice bath to adjust the pH value to 8-9, filtered, and the filter cake was washed with water and dried in vacuum. 4-Chloro-2-methoxyquinazolin-7-yl acetate was obtained as a white solid (9.89 g, 87% yield).

Preparation of Intermediate B:

The preparation method of (S)-4-(chlorocarbonyl)-3-methylpiperazine-1-carboxylic acid tert-butyl ester (intermediate B):

Step f, the preparation of (S)-4-(chlorocarbonyl)-3-methylpiperazine-1-carboxylic acid tert-butyl ester:

At 0°C, under nitrogen protection, pyridine (0.79 g, 10.08 mmol) was added dropwise to a mixture containing 20 mL of anhydrous dichloromethane and trichloromethyl carbonate (1 g, 3.36 mmol), and then (S) - A solution of tert-butyl 3-methylpiperazine-1-carboxylate (0.67g, 3.36mmol) in dichloromethane, after dropping, the mixture was moved to room temperature and stirred overnight. The reaction mixture was concentrated under reduced pressure to obtain yellow solid B (0.79 g, yield 90%), which was directly used in the next reaction without further purification.

Preparation of Intermediate C:

The preparation method of (R)-4-(chlorocarbonyl)-3-methylpiperazine-1-carboxylic acid tert-butyl ester (intermediate C):

Preparation of step g, (R)-4-(chlorocarbonyl)-3-methylpiperazine-1-carboxylic acid tert-butyl ester

At 0°C, under nitrogen protection, pyridine (0.96 g, 12.15 mmol) was added dropwise to a mixture containing 20 mL of anhydrous dichloromethane and trichloromethyl carbonate (1.2 g, 4.05 mmol), and then (R )-3-Methylpiperazine-1-carboxylic acid tert-butyl ester (0.81g, 4.05mmol) in dichloromethane solution, after dropping, the mixture was moved to room temperature and stirred overnight. The reaction mixture was concentrated under reduced pressure to obtain yellow solid C (0.98 g, yield 93%), which was directly used in the next reaction without further purification.

Preparation of Intermediate D:

The preparation method of (R)-4-(chlorocarbonyl)-2-methylpiperazine-1-carboxylic acid tert-butyl ester (intermediate D):

Step h, the preparation of (R)-4-(chlorocarbonyl)-2-methylpiperazine-1-carboxylic acid tert-butyl ester

At 0°C, under nitrogen protection, add pyridine (0.79 g, 10.08 mmol) dropwise to a mixture containing 20 mL of anhydrous dichloromethane and trichloromethyl carbonate (1 g, 3.36 mmol), and then add dropwise (R) - A solution of tert-butyl 2-methylpiperazine-1-carboxylate (0.67g, 3.36mmol) in dichloromethane, after dropping, the mixture was moved to room temperature and stirred overnight. The reaction mixture was concentrated under reduced pressure to obtain yellow solid D (0.78 g, yield 89%), which was directly used in the next reaction without further purification.

Preparation of Intermediate E:

The preparation method of (S)-4-(chlorocarbonyl)-2-methylpiperazine-1-carboxylic acid tert-butyl ester (intermediate E):

The preparation of step i,

At 0°C, under nitrogen protection, pyridine (0.89 g, 11.13 mmol) was added dropwise to a mixture containing 20 mL of anhydrous dichloromethane and trichloromethyl carbonate (1.1 g, 3.71 mmol), and then (S )-2-Methylpiperazine-1-carboxylic acid tert-butyl ester (0.74g, 3.71mmol) in dichloromethane solution, after dropping, the mixture was moved to room temperature and stirred overnight. The reaction mixture was concentrated under reduced pressure to obtain yellow oil E (0.88 g, yield 91%), which was directly used in the next reaction without further purification.

Example 1: 4-(4-(4-chlorophenyl)piperazin-1-yl)-2-methoxyquinazolin-7-yl(R)-4-acryloyl-2-methylpiper Oxyzine-1-carboxylate

The synthetic route is:

Concrete preparation process is as follows:

Preparation of step j, 4-(4-(4-chlorophenyl)piperazin-1-yl)-2-methoxyquinazoline-7-acetate:

Add 1,4-dioxane (15mL) to 4-chloro-2-methoxyquinazoline-7-acetate (500mg, 1.98mmol), add N, N -Diisopropylethylamine (0.27g, 2.15mmol), 1-(4-chlorophenyl)piperazine (0.42g, 2.15mmol), after dropping, stirred at 50°C for 1.5h. The starting material was consumed as monitored by TLC. After standing still, the reaction system was cooled to room temperature, concentrated to dryness, added 20 mL of water, extracted with DCM (2*10 mL), and combined the organic phases. The organic phase was washed with water (1*15mL), washed with saturated sodium chloride (1*10mL), the organic phase was dried by adding anhydrous sodium sulfate, filtered with suction, and the organic phase was distilled under reduced pressure to obtain 4-(4-(4-chlorophenyl )piperazin-1-yl)-2-methoxyquinazoline-7-acetate was directly fed to the next crude product (0.74g, 1.72mmol, crude yield 84%) without further treatment.

Preparation of step k, 4-(4-(4-chlorophenyl)piperazin-1-yl)-2-methoxyquinazolin-7-alcohol:

4-(4-(4-Chlorophenyl)piperazin-1-yl)-2-methoxyquinazoline-7-acetate (0.74 g, 1.79 mmol) was dissolved in anhydrous methanol (10 mL) In, anhydrous potassium carbonate solid (0.53 g, 3.90 mmol) was added, and stirred overnight at room temperature. TLC detects that the reaction is complete. Distilled under reduced pressure to obtain a residue purified by column chromatography to obtain 4-(4-(4-chlorophenyl)piperazin-1-yl)-2-methoxyquinazolin-7-ol (0.463g, 1.25 mmol, yield 70%).

Step 1, 4-(tert-butyl)1-(4-(4-(4-chlorophenyl)piperazin-1-yl)-2-methoxyquinazolin-7-yl)(R)- Preparation of 2-methylpiperazine-1,4-dicarboxylate:

(R)-tert-butyl 3-methylpiperazine-1-carboxylate (0.5 g, 2.5 mmol) was added to 4-(4-(4-chlorophenyl)piperazin-1-yl)-2 -Methoxyquinazolin-7-ol (0.463g, 1.25mmol), and anhydrous potassium carbonate (0.345g, 2.5mmol) in the reaction system of N,N-dimethylformamide (13mL), room temperature Stir overnight. TLC detects that the reaction of raw materials is complete. Add the reaction solution dropwise to 60mL of ice water which is constantly stirring, stir for 10min after the dropwise addition, let stand for 10min, filter with suction, dry the filter cake in a vacuum oven at 40°C for 3h, and obtain 4-(tert-butyl) 1-(4-(4-(4-chlorophenyl)piperazin-1-yl)-2-methoxyquinazolin-7-yl)(R)-2-methylpiperazine-1,4 -Dicarboxylate (483 mg, 0.81 mmol, yield 65.8%).

Step m, 4-(4-(4-chlorophenyl)piperazin-1-yl)-2-methoxyquinazolin-7-yl(R)-2-methylpiperazine-1-carboxylic acid Preparation of esters

In compound 4-(tert-butyl)1-(4-(4-(4-chlorophenyl)piperazin-1-yl)-2-methoxyquinazolin-7-yl)(R)-2 -Methylpiperazine-1,4-dicarboxylate (483 mg, 0.81 mmol) was added with 15 mL of 4M hydrogen chloride-methanol solution, stirred at room temperature for 1 h, and the reaction of raw materials was detected by TLC. Distillation under reduced pressure gives compound 4-(4-(4-chlorophenyl) piperazin-1-yl)-2-methoxyquinazolin-7-yl (R)-2-methylpiperazine-1- Carboxylate hydrochloride (0.38g, 0.78mmol, yield 97%) was directly used in the next reaction without further treatment.

Step n, 4-(4-(4-chlorophenyl)piperazin-1-yl)-2-methoxyquinazolin-7-yl(R)-4-acryloyl-2-methylpiperazine - Preparation of 1-carboxylate

-20°C, under nitrogen atmosphere, to compound 4-(4-(4-chlorophenyl)piperazin-1-yl)-2-methoxyquinazolin-7-yl(R)-2-methyl Piperazine-1-carboxylate hydrochloride (0.38g, 0.78mmol), add dichloromethane (8.0mL), N,N-diisopropylethylamine (403mg, 3.12mmol), dissolve and add di A solution of acryloyl chloride (211 mg, 2.34 mmol) diluted with methyl chloride (0.5 mL) was stirred at -20°C for 30 min. TLC detects that the reaction of the raw materials is complete, the reaction solution is moved to room temperature, and 10 mL of saturated sodium bicarbonate and 3 mL are added and stirred for 30 min, separated, the organic phase is washed with water (2*10 mL), and saturated sodium chloride solution (2*10 mL), After the organic phase was evaporated to dryness, the sample was directly mixed and passed through the column to obtain 4-(4-(4-chlorophenyl)piperazin-1-yl)-2-methoxyquinazolin-7-yl(R)-4- Acryloyl-2-methylpiperazine-2-carboxylate (236 mg, 0.43 mmol, yield 56.3%).

MS (ESI) m/z: 551 [M+H] ⁺ . ¹ H NMR (400MHz, Chloroform-d) δ8.69(s, 1H), 7.62(s, 1H), 7.28(d, J=19.1Hz, 1H), 6.99(t, J=8.6Hz, 2H), 6.92(dd,J=9.1,4.6Hz,2H),6.56(t,J=13.6Hz,1H),6.41–6.33(m,1H),5.80–5.73(m,1H),4.64(s,1H) ,3.96(s,3H),3.90–3.83(m,4H),3.29(s,10H),1.31(s,3H).

The corresponding substituted quinazoline compounds of Examples 2-7 were synthesized according to the operation similar to that of Example 1.

Example 2: 2-methoxy-4-(4-phenylpiperazin-1-yl)quinazolin-7-yl (R)-4-acryloyl-2-methylpiperazine-1-carboxy Ester

Synthesis of 2-methoxy-4-(4-phenylpiperazin-1-yl)quinazolin-7-yl(R)-4-acryloyl-2-methylpiperazine-1-carboxylate Refer to Example 1. LC-MS (ESI) m/z: 517 [M+H] ⁺ . ¹ H NMR (400MHz, DMSO-d ₆ )δ8.62(s, 1H), 7.75(s, 1H), 7.35(s, 1H), 7.24(t, J=7.9Hz, 2H), 6.98(d, J＝8.1Hz, 2H), 6.79(d, J＝7.1Hz, 1H), 6.22–6.13(m, 1H), 5.75(s, 1H), 5.73(d, J＝2.2Hz, 1H), 4.32( d,J＝38.4Hz,1H),3.94(s,3H),3.83–3.80(m,4H),3.35(s,10H),1.24(s,3H).

Example 3: 2-methoxy-4-(4-phenylpiperazin-1-yl)quinazolin-7-yl (S)-4-acryloyl-2-methylpiperazine-1-carboxy Ester

Synthesis of 2-methoxy-4-(4-phenylpiperazin-1-yl)quinazolin-7-yl(S)-4-acryloyl-2-methylpiperazine-1-carboxylate Refer to Example 1. LC-MS (ESI) m/z: 517 [M+H] ⁺ . ¹ H NMR (400MHz, DMSO-d ₆ )δ8.70(s, 1H), 7.64(s, 1H), 7.31(s, 1H), 7.24(t, J=7.9Hz, 2H), 6.99(d, J＝8.1Hz, 2H), 6.89(d, J＝7.1Hz, 1H), 6.22–6.13(m, 1H), 5.75(s, 1H), 5.73(d, J＝2.2Hz, 1H), 4.32( d,J＝38.4Hz,1H),3.94(s,3H),3.83–3.80(m,4H),3.39(s,10H),1.77(s,3H).

Example 4: 4-(4-(4-chlorophenyl)piperazin-1-yl)-2-methoxyquinazolin-7-yl(R)-4-acryloyl-3-methylpiper Oxyzine-1-carboxylate

4-(4-(4-Chlorophenyl)piperazin-1-yl)-2-methoxyquinazolin-7-yl(R)-4-acryloyl-3-methylpiperazine-1- The synthesis of carboxylate refers to Example 1. MS (ESI) m/z: 551 [M+H] ⁺ . ¹ H NMR (400MHz, Chloroform-d) δ8.68(s,1H),7.62(s,1H),7.40(s,1H),7.26–7.21(m,2H),6.87(d,J＝8.9Hz ,2H),6.40(s,1H),6.17–6.05(m,1H),5.88–5.82(m,1H),5.80–5.75(m,1H),3.96(s,3H),3.92–3.87(m ,4H),3.33(d,J＝9.1Hz,8H),2.94–2.66(m,2H),1.32(s,3H).

Example 5: 4-(4-(4-chlorophenyl)piperazin-1-yl)-2-methoxyquinazolin-7-yl(S)-4-acryloyl-3-methylpiper Oxyzine-1-carboxylate

4-(4-(4-Chlorophenyl)piperazin-1-yl)-2-methoxyquinazolin-7-yl(S)-4-acryloyl-3-methylpiperazine-1- The synthesis of carboxylate refers to Example 1. MS (ESI) m/z: 551 [M+H] ⁺ . ¹ H NMR (400MHz, Chloroform-d) δ8.70(s, 1H), 7.62(d, J=6.6Hz, 1H), 7.32(s, 1H), 7.28–7.21(m, 2H), 6.88(d ,J=8.6Hz,2H),6.58(dd,J=16.3,10.7Hz,1H),6.35(d,J=16.6Hz,1H),5.76(d,J=10.5Hz,1H),4.15(dd ,J=17.6,10.0Hz,1H),3.96(s,3H),3.87(s,4H),3.42–2.99(m,8H),2.05(s,2H),1.37(d,J=14.9Hz, 3H).

Example 6: 2-methoxy-4-(4-(4-methoxyphenyl)piperazin-1-yl)quinazolin-7-yl(R)-4-acryloyl-2-methyl Piperazine-1-carboxylate

2-Methoxy-4-(4-(4-methoxyphenyl)piperazin-1-yl)quinazolin-7-yl(R)-4-acryloyl-2-methylpiperazine- The synthesis of 1-carboxylate refers to Example 1. MS (ESI) m/z: 547 [M+H] ⁺ . ¹ H NMR (400MHz, Chloroform-d) δ8.69(s, 1H), 7.63(d, J=6.9Hz, 1H), 7.32(s, 1H), 7.29–7.18(m, 3H), 6.89(d ,J=8.7Hz,2H),6.59(d,J=16.9Hz,1H),6.35(d,J=11.5Hz,1H),5.74(d,J=4.8Hz,1H),3.96(s,6H ),3.86(s,4H),3.34(s,10H),1.36(d,J=19.9Hz,3H).

Example 7: KRAS ^G12C -SOS1 protein complex inhibition rate test

The testing concentration of the compound to be tested is 500 nM, and the 100% DMSO solution diluted to 200-fold final concentration in a 384-well plate. Use the dispenser Echo 550 to transfer 50 nl of the compound with a final concentration of 200 times to the target plate 384-well-plate. Add 50 nl of 100% DMSO to the negative control wells and positive control wells respectively, prepare a 4-fold final concentration of Tag1-SOS1 solution with Diluent buffer, add 2.5 μl of 4-fold final concentration of Tag1-SOS1 solution to a 384-well plate, and use Prepare a Tag2-KRAS-G12C solution with 4 times the final concentration in diluent buffer; add 2.5 μl of diluent buffer to the negative control well. The 384-well plate was centrifuged at 1000 rpm for 30 seconds, shaken to mix and incubated at room temperature for 15 minutes. Anti-Tag1-TB3+ solution with 1 times final concentration and Anti-Tag2-XL665 solution with 1 times final concentration were prepared in Detection buffer. After mixing the two solutions, add 5 μl of Mix solution to each well. The 384-well plate was centrifuged at 1000rpm for 30s, shaken and mixed, incubated at room temperature for 120min, and the reading was recorded with an Envision microplate reader. The specific results are shown in Table 1.

The specific inhibitory rate (500nM) of table 1 embodiment and commercially available compound

Example number Inhibition rate of KRAS^G12C-SOS1 protein complex (n=2) 1 60.7%±3.2 2 45.7%±1.2 3 41.2%±0.7 4 63.1%±3.2 5 63.2%±3.5 6 93.8%±4.2 ARS-1620 81.7%±1.2

While the invention has been illustrated by the foregoing specific examples, it should not be construed as being limited thereto; rather the invention encompasses the general aspects disclosed previously. Various modifications can be made and have various embodiments without departing from the spirit and scope of the invention.

Claims (10)

1. A substituted quinazoline compound with a structural general formula shown as formula 1, or optical isomers, crystal forms, pharmaceutically acceptable salts, hydrates or solvates thereof,

wherein:

m is 1 or 2;

R ¹ selected from methoxy, hydroxy, amino, halogen, methylamino, C1-C2 alkyl, C1-C2 alkoxy;

y is oxygen or nitrogen;

R ² selected from aryl or heteroaryl, substituted or unsubstituted; wherein aryl or heteroaryl is substituted with 1-2 of the following groups: halogen, C1-C2 alkoxy, hydroxy, amino, cyano, C1-C2 alkyl, C3-C6 cycloalkyl;

R ³ is mono-to tri-substituted and is selected from hydrogen,Methyl, hydroxymethyl, or

Is composed of

R ⁴ Independently selected from hydrogen, fluorine, methoxy;

R ⁵ is mono-to tri-substituted and is selected from hydrogen, methyl, hydroxymethyl; or alternatively

Is composed of

2. The substituted quinazoline compound according to claim 1, or each optical isomer, each crystal form, pharmaceutically acceptable salt, hydrate or solvate thereof, wherein the aryl group is a substituted or unsubstituted benzene ring or naphthalene ring.

3. The substituted quinazoline compound according to claim 1, or each optical isomer, each crystal form, pharmaceutically acceptable salt, hydrate or solvate thereof, wherein the heteroaryl group means that the substituted or unsubstituted heteroaryl group is a mono-or bicyclic aromatic ring system having 5 to 12 ring atoms, wherein at least one atom in the ring system is a heteroatom selected from N, O and S.

4. The substituted quinazoline compound according to claim 1, or each optical isomer, each crystal form, pharmaceutically acceptable salt, hydrate or solvate thereof, wherein the pharmaceutically acceptable salt is an inorganic salt or an organic salt; the inorganic salt comprises hydrochloride, hydrobromide, hydroiodide, sulfate, bisulfate, nitrate, phosphate and acid phosphate; the organic salt is selected from acetate, trifluoroacetate, propionate, pyruvate, glycolate, oxalate, malonate, fumarate, maleate, lactate, malate, citrate, tartrate, methanesulfonate, benzenesulfonate, salicylate.

5. Use of the substituted quinazoline compound according to any one of claims 1 to 4, or each optical isomer, each crystal form, a pharmaceutically acceptable salt, a hydrate or a solvate thereof, in the preparation of a medicament for preventing or treating KRAS ^G12C Use in the manufacture of a medicament for a protein mediated disease.

6. A pharmaceutical composition comprising the substituted quinazoline compound according to any one of claims 1 to 4, or each optical isomer, each crystal form, a pharmaceutically acceptable salt, a hydrate or a solvate thereof.

7. The pharmaceutical composition of claim 6, further comprising: a pharmaceutically acceptable carrier, excipient or diluent.

8. Use of the substituted quinazoline compound of any one of claims 1 to 4, or the pharmaceutical composition of the optical isomers, crystal forms, pharmaceutically acceptable salts, hydrates or solvates thereof for the preparation of KRAS ^G12C Use in mutation inhibitors.

9. A medicament for treating cancer, which comprises a pharmaceutical composition of the substituted quinazoline compound as claimed in any one of claims 1 to 4, or each optical isomer, each crystal form, a pharmaceutically acceptable salt, a hydrate or a solvate thereof.

10. The medicament for treating cancer according to claim 9, wherein the cancer is ductal carcinoma of pancreas, colorectal cancer, multiple myeloma, lung cancer, cutaneous melanoma, endometrioid carcinoma of uterus, sarcoma of uterus cancer, thyroid cancer, acute myelogenous leukemia, urothelial carcinoma of bladder, stomach cancer, cervical cancer, squamous cell carcinoma of head and neck, diffuse large B-cell lymphoma, esophageal cancer, chronic lymphocytic leukemia, squamous cell carcinoma of lung, small cell lung cancer, papillary cell carcinoma of kidney, adenoid cystic carcinoma, chromophobe renal cell carcinoma, liver cancer, breast infiltrating cancer, squamous cell carcinoma of cervix, serous adenocarcinoma of ovary, adrenocortical carcinoma, prostate cancer, neuroblastoma, brain low-grade glioma, glioblastoma, medulloblastoma, esophageal cell carcinoma, renal clear cell squamous cell carcinoma, osteosarcoma, small cell ovarian cancer, rhabdoid tumor, sarcoma, small-intestinal neuroblastoma, T-cell prolymphocytic leukemia.