WO2017092413A1

WO2017092413A1 - Diaminopyrimidine compounds and composition comprising same

Info

Publication number: WO2017092413A1
Application number: PCT/CN2016/096320
Authority: WO
Inventors: 王义汉; 李焕银
Original assignee: 深圳市塔吉瑞生物医药有限公司
Priority date: 2015-12-02
Filing date: 2016-08-23
Publication date: 2017-06-08
Also published as: CN106188138A; CN106188138B; CN108948082A; CN113912648A

Abstract

Provided are diaminopyrimidine compounds as shown in formula (I), and a pharmaceutical composition comprising the compounds or a crystal form, a pharmaceutically acceptable salt, a hydrate or solvate, a stereoisomer, a prodrug or an isotopic variant thereof. The diaminopyrimidine compounds and the composition comprising the compounds exhibit an excellent inhibitory effect against protein kinases. They also have better pharmacokinetic parameter characteristics, which allows increasing drug concentration of the compounds in an animal body and enhances drug efficacy and safety.

Description

Diaminopyrimidine compound and composition comprising the same

Technical field

The invention belongs to the technical field of medicine, and in particular relates to a diaminopyrimidine compound and a composition comprising the same.

Background technique

In the past 30 years, lung cancer mortality has increased by 465%, and the incidence rate has increased by 26.9% per year, which has become the cause of the first malignant tumor death in China. Non-small cell lung cancer (NSCLC) accounts for more than 80% of all lung cancers. Only one-third of patients with NSCLC have a chance of surgery. About 70% of patients have a local advanced stage at the time of presentation. Or there is a distant metastasis and the chance of surgery is lost. In this case, drug treatment is particularly important. The anaplastic lymphoma kinase (ALK) gene fusion has recently become an important biomarker for patients with specific NSCLC subgroups, and thus with the corresponding inhibitors. The International Association for the Study of Lung Cancer (IASLC) recommends ALK fusion testing to guide patient screening, and patients with advanced adenocarcinoma who are eligible for ALK inhibitors regardless of gender, race, smoking history, or other clinical risk factors. Fluorescence in situ hybridization (FISH) with dual-tag separation probes is used to select patients who are eligible for ALK-TKI therapy. This diagnostic method has been approved by the US FDA and has been used in the study of crizotinib for the treatment of ALK rearranged tumors. Adopted. Crizotinib is a competitive inhibitor of oral adenosine triphosphate (ATP) that inhibits ALK and MET tyrosine kinases and also inhibits the activity of ROS1 and RON kinases.

However, crizotinib has the following side effects: visual impairment, gastrointestinal side effects, and elevated levels of grade 3-4 liver transaminases in 16% of cases. In addition, ALK-positive patients inevitably acquire acquired resistance after the initial phase of crizotinib treatment. Thus, there is a need to develop compounds that have ALK kinase inhibitory activity and/or have better pharmacodynamic/pharmacokinetic properties.

Summary of the invention

In view of the above technical problems, the present invention discloses a diaminopyrimidine compound and a composition comprising the same which has better ALK kinase inhibitory activity and/or has better pharmacodynamic/pharmacokinetic properties.

In this regard, the technical solution adopted by the present invention is:

It is an object of the present invention to provide a novel class of compounds having ALK kinase inhibitory activity and/or having better pharmacodynamic/pharmacokinetic properties.

In a first aspect of the invention, there is provided a diaminopyrimidine compound of the formula (I), or a crystalline form thereof, a pharmaceutically acceptable salt, a hydrate or a solvent compound.

In the formula:

Wherein: R ^1a , R ^1b , R ^1c , R ^2a , R ^2b , R ^3a , R ^3b , R ^4a , R ^4b , R ^5a , R ^5b , R ⁶ , R ^7a , R ^7b , R ^8a , R ^8b , R ^9a , R ^9b , R ^10a , R ^10b , R ¹¹ , R ¹² , R ¹³ , R ^14a , R ^14b , R ^14c , R ¹⁵ , R ¹⁶ , R ^17a , R ^17b , R ^17c , R ^18a , R ^18b And R ^18c , R ¹⁹ , R ²⁰ , R ²¹ and R ²² are each independently hydrogen, deuterium, halogen or trifluoromethyl;

R ¹⁶ is hydrogen, deuterium, halogen, cyano, unsubstituted C ₁ -C ₆ alkyl or C ₁ -C ₆ alkoxy, one or more deuterated or fully deuterated C ₁ -C ₆ An alkyl or C ₁ -C ₆ alkoxy group, or one or more halogen-substituted or perhalogen-substituted C ₁ -C ₆ alkyl or C ₁ -C ₆ alkoxy;

Additional conditions are R ^1a , R ^1b , R ^1c , R ^2a , R ^2b , R ^3a , R ^3b , R ^4a , R ^4b , R ^5a , R ^5b , R ⁶ , R ^7a , R ^7b , R ^8a , R ^8b And R ^9a , R ^9b , R ^10a , R ^10b , R ¹¹ , R ¹² , R ¹³ , R ^14a , R ^14b , R ^14c , R ¹⁵ , R ¹⁶ , R ^17a , R ^17b , R ^17c , R ^18a , R At least one of ^18b , R ^18c , R ¹⁹ , R ²⁰ , R ²¹ and R ²² is deuterated or deuterated.

The shape and volume of hydrazine in the drug molecule are substantially the same as hydrogen. If the hydrogen in the drug molecule is selectively replaced with hydrazine, the deuterated drug generally retains its original biological activity and selectivity. At the same time, the inventors have confirmed through experiments that the binding of carbon-germanium bonds is more stable than the combination of carbon-hydrogen bonds, which can directly affect the absorption, distribution, metabolism and excretion of some drugs, thereby improving the efficacy, safety and tolerability of the drugs.

In another preferred embodiment, the strontium isotope content of strontium at each metamorphic position is at least greater than the natural strontium isotope content (0.015%), preferably greater than 30%, more preferably greater than 50%, and even more preferably greater than 75%. More preferably greater than 95%, more preferably greater than 99%.

Specifically, in the present invention, R ^1a , R ^1b , R ^1c , R ^2a , R ^2b , R ^3a , R ^3b , R ^4a , R ^4b , R ^5a , R ^5b , R ⁶ , R ^7a , R ^7b , R ^8a , R ^8b , R ^9a , R ^9b , R ^10a , R ^10b , R ¹¹ , R ¹² , R ¹³ , R ^14a , R ^14b , R ^14c , R ¹⁵ , R ¹⁶ , R ^17a , R ^17b , R ^17c And 氘 isotope content in each of the deuterated positions of R ^18a , R ^18b , R ^18c , R ¹⁹ , R ²⁰ , R ²¹ and R ²² is at least 5%, preferably more than 10%, more preferably more than 15%, more preferably More than 20%, more preferably more than 25%, more preferably more than 30%, more preferably more than 35%, more preferably more than 40%, more preferably more than 45%, more preferably more than 50%, more preferably More than 55%, more preferably more than 60%, more preferably more than 65%, more preferably more than 70%, more preferably more than 75%, more preferably more than 80%, more preferably more than 85%, more preferably more than 90%, more preferably greater than 95%, and even more preferably greater than 99%.

In another preferred embodiment, the compound of the formula (I) contains at least one deuterium atom, and the number of deuterium atoms may be any one of 1 to 38.

In another preferred embodiment, R ^1a , R ^1b , R ^1c , R ^2a , R ^2b , R ^3a , R ^3b , R ^4a , R ^4b , R ^5a , R ^5b , R ⁶ of the compound of formula (I), R ^7a , R ^7b , R ^8a , R ^8b , R ^9a , R ^9b , R ^10a , R ^10b , R ¹¹ , R ¹² , R ¹³ , R ^14a , R ^14b , R ^14c , R ¹⁵ , R ¹⁶ , R ^17a And R ^17b , R ^17c , R ^18a , R ^18b , R ^18c , R ¹⁹ , R ²⁰ , R ²¹ and R ²² , at least one of R contains ruthenium, more preferably two R contains ruthenium, more preferably three R氘, preferably four R 氘, more preferably five R 氘, more preferably six R 氘, more preferably seven R 氘, more preferably eight R 氘, better Nine R 氘, preferably ten R 氘, more preferably eleven R 氘, more preferably twelve R 氘, more preferably thirteen R 氘, better ten The four Rs contain ruthenium, more preferably fifteen R 氘, more preferably sixteen R 氘, more preferably seventeen R 氘, more preferably eighteen R 氘, more preferably ten Nine R containing bismuth, more preferably twenty R 氘, more preferably twenty-one R containing 氘, more preferably twenty-two R containing 氘, more preferably twenty-three R containing 氘, more Jiadi 20 R contains 氘, more preferably twenty-five R contains 氘, more preferably twenty-six R contains 氘, more preferably twenty-seven R contains 氘, more preferably twenty-eight R contains 氘, more Good land twenty-nine R contains 氘, more preferably thirty R contains 氘, more preferably thirty-one R contains 氘, more preferably thirty-two R contains 氘, more preferably thirty-three R氘, preferably thirty-four R 氘, more preferably thirty-five R 氘, more preferably thirty-six R 氘, more preferably thirty-seven R 氘, more preferably Thirty-eight R contains 氘.

In another preferred embodiment, R ^1a , R ^1b and R ^1c are each independently hydrazine or hydrogen.

In another preferred embodiment, R ^2a , R ^2b , R ^3a , R ^3b , R ^4a , R ^4b , R ^5a and R ^5b are each independently hydrazine or hydrogen.

In another preferred embodiment, R ⁶ is deuterium or hydrogen.

In another preferred embodiment, R ^7a , R ^7b , R ^8a , R ^8b , R ^9a , R ^9b , R ^10a and R ^10b are each independently hydrazine or hydrogen.

In another preferred embodiment, R ¹¹ , R ¹² and R ¹³ are each independently hydrazine or hydrogen.

In another preferred embodiment, R ^14a, R ^14b and R ^14c are each independently hydrogen or deuterium.

In another preferred embodiment, R ^17a , R ^17b and R ^17c are each independently hydrazine or hydrogen.

In another preferred embodiment, R ^18a , R ^18b and R ^18c are each independently hydrazine or hydrogen.

In another preferred embodiment, R ¹⁹ , R ²⁰ , R ²¹ and R ²² are each independently hydrazine or hydrogen.

In another preferred embodiment, R ^{16 is} independently selected from the group consisting of halogen, trifluoromethyl, cyano, one or more deuterated alkyl groups and alkoxy groups.

In another preferred embodiment, the compound is selected from the group consisting of the compounds or pharmaceutically acceptable salts thereof:

In another preferred embodiment, the compound does not include a non-deuterated compound.

In another preferred embodiment, the non-deuterated compound is 5-chloro-N4-(2-(dimethylphosphinyl)phenyl)-N2-(2-methoxy-4-(4- (4-Methylpiperazin-1-yl)-piperidin-1-yl)phenyl)pyrimidine-2,4-diamine.

In a second aspect of the invention, there is provided a method of preparing a pharmaceutical composition comprising the steps of: pharmaceutically acceptable carrier and a compound of the first aspect of the invention, or a crystalline form thereof, pharmaceutically acceptable The accepted salt, hydrate or solvate is mixed to form a pharmaceutical composition.

In a third aspect of the invention, there is provided a pharmaceutical composition comprising a pharmaceutically acceptable carrier and a compound of the first aspect of the invention, or a crystalline form thereof, a pharmaceutically acceptable salt, hydrated Or a solvate.

In another preferred embodiment, the pharmaceutical composition is an injection, a sachet, a tablet, a pill, a powder or a granule.

In another preferred embodiment, the pharmaceutical composition further comprises an additional therapeutic agent, which is cancer, cardiovascular disease, inflammation, infection, immune disease, cell proliferative disease, viral disease. , a metabolic disease, or a drug for organ transplantation.

In a fourth aspect of the invention, the compound of the first aspect of the invention, or a crystal form thereof, or a drug thereof The use of a physiologically acceptable salt, prodrug, stereoisomer, isotopic variation, hydrate or solvate for the preparation of a pharmaceutical composition for inhibiting protease.

In another preferred embodiment, the pharmaceutical composition is for treating and preventing diseases such as cancer, cell proliferative diseases, inflammation, infection, immune diseases, organ transplantation, viral diseases, cardiovascular diseases or metabolic diseases. .

In another preferred embodiment, the cancer includes, but is not limited to, lung cancer, head and neck cancer, breast cancer, prostate cancer, esophageal cancer, rectal cancer, colon cancer, nasopharyngeal cancer, uterine cancer, pancreatic cancer, lymphoma, Blood cancer, osteosarcoma, melanoma, kidney cancer, stomach cancer, liver cancer, bladder cancer, thyroid cancer or colorectal cancer.

In another preferred embodiment, the immune disease or inflammation includes, but is not limited to, rheumatoid arthritis, osteoarthritis, rheumatoid spondylitis, gout, asthma, bronchitis, rhinitis, chronic obstructive pulmonary disease, Cystic fibrosis.

In another preferred embodiment, the cell proliferative disorder refers to lung cancer, head and neck cancer, breast cancer, prostate cancer, esophageal cancer, rectal cancer, colon cancer, nasopharyngeal cancer, uterine cancer, pancreatic cancer, lymphoma, blood cancer. , osteosarcoma, melanoma, kidney cancer, stomach cancer, liver cancer, bladder cancer, thyroid cancer or colorectal cancer.

In another preferred embodiment, the cancer is non-small cell lung cancer.

In a fifth aspect of the invention, there is provided a method of inhibiting a protein kinase (such as ALK kinase) or a disease (such as cancer, cell proliferative disease, inflammation, infection, immune disease, organ transplantation, viral disease) A method of treating a cardiovascular disease or a metabolic disease comprising the step of administering a compound described in the first aspect of the invention, or a crystalline form, a pharmaceutically acceptable salt, a hydrate or a solvent thereof, to a subject in need of treatment The composition, or the pharmaceutical composition described in the third aspect of the invention.

It is to be understood that within the scope of the present invention, the various technical features of the present invention and the various technical features specifically described hereinafter (as in the embodiments) may be combined with each other to constitute a new or preferred technical solution. Due to space limitations, we will not repeat them here.

The invention also includes isotopically labeled compounds, equivalent to the original compounds disclosed herein. Examples of isotopes which may be listed as compounds of the present invention include hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine and chlorine isotopes such as ² H, ³ H, ¹³ C, ¹⁴ C, ¹⁵ N, ¹⁷ O, ¹⁸ O, respectively. , ³¹ P, ³² P, ³⁵ S, ¹⁸ F and ³⁶ Cl. a compound, or an enantiomer, a diastereomer, an isomer, or a pharmaceutically acceptable salt or solvate of the present invention, wherein an isotope or other isotopic atom containing the above compound is within the scope of the present invention . Certain isotopically-labeled compounds of the present invention, such as the radioisotopes of ³ H and ¹⁴ C, are also among them, useful in tissue distribution experiments of drugs and substrates.氚, ie ³ H and carbon-14, ie ¹⁴ C, are easier to prepare and detect and are preferred in isotopes. In addition, heavier isotopic substitutions such as guanidine, or ² H, are preferred in certain therapies due to their good metabolic stability, such as increased half-life or reduced dosage in vivo, and therefore may be preferred in certain circumstances. Isotopically labeled compounds can be prepared in a conventional manner by replacing the readily available isotopically labeled reagent with a non-isotopic reagent using the protocol of the examples.

Herein, "halogen" means F, Cl, Br, and I unless otherwise specified. More preferably, the halogen atom is selected from the group consisting of F, Cl and Br.

Herein, "C ₁ -C ₆ alkyl" means a straight or branched alkyl group having 1 to 6 carbon atoms, such as methyl, ethyl, propyl, isopropyl, unless otherwise specified. Butyl, isobutyl, tert-butyl, or the like.

As used herein, unless otherwise specified, "deuterated" means that one or more hydrogens in the compound or group are replaced by deuterium; deuteration may be monosubstituted, disubstituted, polysubstituted or fully substituted. The terms "one or more deuterated" are used interchangeably with "one or more deuterated".

As used herein, unless otherwise specified, "non-deuterated compound" means a compound containing a proportion of germanium atoms not higher than the natural helium isotope content (0.015%).

In the present invention, pharmaceutically acceptable salts include inorganic salts and organic salts. A preferred class of salts are the salts of the compounds of the invention with acids. Suitable acids for forming salts include, but are not limited to, mineral acids such as hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid, phosphoric acid; formic acid, acetic acid, trifluoroacetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, Organic acids such as fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, picric acid, benzoic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, benzenesulfonic acid, naphthalenesulfonic acid; Amino acids such as amino acid, phenylalanine, aspartic acid, and glutamic acid. Another preferred class of salts are the salts of the compounds of the invention with bases, such as alkali metal salts (for example sodium or potassium salts), alkaline earth metal salts (for example magnesium or calcium salts), ammonium salts (for example lower alkanolammonium). Salts and other pharmaceutically acceptable amine salts), such as methylamine, ethylamine, propylamine, dimethylamine, trimethylamine, diethylamine, triethylamine, tert-butyl A base amine salt, an ethylenediamine salt, a hydroxyethylamine salt, a dihydroxyethylamine salt, a trihydroxyethylamine salt, and an amine salt formed of morpholine, piperazine, and lysine, respectively.

The term "solvate" refers to a complex of a compound of the invention that is coordinated to a solvent molecule to form a specific ratio. "Hydrate" means a complex formed by the coordination of a compound of the invention with water.

Compared with the prior art, the beneficial effects of the present invention are: the compound of the present invention has excellent inhibitory effect on a protein kinase (for example, ALK kinase); the technology of sputum is used to change the metabolism of the compound in the organism, so that The compounds have better pharmacokinetic parameter properties. In this case, the dosage can be changed and a long-acting preparation can be formed to improve the applicability; replacing the hydrogen atom in the compound with hydrazine can increase the drug concentration of the compound in the animal due to its strontium isotope effect, thereby improving the therapeutic effect; The substitution of a hydrogen atom in a compound by hydrazine may increase the safety of the compound due to inhibition of certain metabolites.

detailed description

The preparation of the structural compound of the formula (I) of the present invention is more specifically described below, but these specific methods do not constitute any limitation to the present invention. The compounds of the invention may also optionally be described in the specification or known in the art. Such synthetic methods are conveniently combined and can be readily made by those skilled in the art to which the present invention pertains.

The preparation of the unsubstituted diaminopyrimidine compound and its physiologically compatible salt used in the present invention is known. The preparation of the diaminopyrimidine compound corresponding to the deuterated can be carried out by the same route using the corresponding deuterated starting compound as a starting material. For example, the compound of the formula (I) of the present invention can be produced according to the preparation method described in WO2012061299, except that the non-deuterated raw material is replaced with a deuterated raw material in the reaction. Usually, in the preparation scheme, each reaction is usually carried out in an inert solvent at room temperature to reflux temperature (e.g., 0 ° C to 100 ° C, preferably 0 ° C to 80 ° C). The reaction time is usually from 0.1 to 60 hours, preferably from 0.5 to 24 hours.

The following general preparative routes can be used to synthesize the compounds of the formula (I) of the present invention. The synthetic route is as follows:

The synthesis of piperazine substituted piperidinamide is as follows:

The details will be described below in conjunction with the embodiments.

Example 1

According to the following Scheme Preparation of 5-chloro -N ⁴ - [2- (dimethylphosphoryl) phenyl] -N ² - ^{{2-d3--methoxy-4-} [4- (4-methylpiperazine -1-yl)-piperidin-1-yl]phenyl}pyrimidine-2,4-diamine (compound 9):

(1) Preparation of Compound 2: 30 mL of acetone was added to a 100 mL single-mouth bottle, and 5-fluoro-2-nitrophenol (2.0 g, 12.7 mmol), anhydrous potassium carbonate (3.5 g, 25.4 mmol), and hydrazine were sequentially added thereto with stirring. Methyl iodide (2.4 g, 16.5 mmol) was warmed to 60 ° C and stirred for 2 h. The mixture was cooled to room temperature, and the residue was evaporated to dryness. ^{_{1 H NMR (300MHz, CDCl 3}} ) (δ / ppm) 8.00-7.95 (m, 1H), 6.83-6.71 (m, 2H), LC-MS (APCI): m / z = 175.2 (M + 1) + , 95%.

(2) Preparation of Compound 4: N,N-dimethylformamide (4 mL) was added to a 25 mL single-necked flask, and 4-fluoro-2-d3-methoxynitrobenzene (0.4 g, 2.3 mmol) was sequentially added with stirring. , 1-methyl-4-(piperidin-4-yl)piperazine hydrochloride (0.7 g, 3.2 mmol), anhydrous potassium carbonate (0.95 g, 6.9 mmol), and the mixture was warmed to 80 ° C, N ₂ reactions overnight. After cooling to room temperature, poured into ice water (80 mL), EtOAc (EtOAc:EtOAc) LC-MS (APCI): m / z = 338.2 (M + 1) +; 1 H NMR (300MHz, CDCl 3) (δ / ppm) 8.01 (d, J = 9.3Hz, 1H), 6.43 (dd, J = 9.6 Hz, J = 2.7 Hz, 1H), 6.31 (d, J = 2.4 Hz, 1H), 3.98 - 3.94 (m, 2H), 3.03 - 2.94 (m, 2H), 2.65 - 2.62 (m, 4H) , 2.54-2.46 (m, 5H), 2.32 (s, 3H), 2.01-1.96 (m, 2H), 1.65-1.60 (m, 2H).

(3) Preparation of compound 5: 6 mL of ethanol and 2 mL of water were added to a 25 mL single-mouth bottle, and 1-(1-(3-d3-methoxy-4-nitrophenyl)piperidin-4-yl was added in sequence with stirring. -4-methylpiperazine (0.2 g, 0.59 mmol), reduced iron powder (0.20 g, 3.55 mmol), ammonium chloride (16 mg, 0.30 mmol), the reaction mixture was heated to 85 ° C under N ₂ atmosphere, and kept warm The reaction was stirred for 1 h. The mixture was cooled to room temperature, the solid was filtered, evaporated, evaporated, evaporated, evaporated, evaporated, evaporated The solid was 0.15 g, the yield was 79.6%, and it was directly poured into the next step.

(4) Preparation of Compound 8: DMF (3 mL) was added to a 25 mL single-necked flask, and 2,5,6-trichloropyrimidine (0.72 g, 3.9 mmol) and 2-(dimethylphosphine oxide) aniline were added in sequence with stirring. 0.5 g, 3 mmol), anhydrous potassium carbonate (0.62 g, 4.5 mmol), warmed to 60 ° C and stirred for 4 h. After cooling to room temperature, ethyl acetate (30 mL) and water (30 mL) were added successively, and the layers were separated, and the aqueous layer was extracted with ethyl acetate (30 mL x 2). The organic phase was combined, washed with water (60 mL x 2) The residue was dried over sodium sulfate, filtered and evaporated. LC-MS (APCI): m / z = 175.2 (M + 1) +; 1 H NMR (CDCl 3, 300MHz) (δ / ppm) 8.00-7.95 (m, 1H), 6.83-6.71 (m, 2H) .

(5) Preparation of compound 9: 2 mL of ethylene glycol monomethyl ether was added to a 25 mL single-mouth bottle, and 2,5-dichloro-N-(2-(dimethyloxyphosphino)phenyl)pyrimidine was sequentially added under stirring. 4-amine (60 mg, 0.19 mmol), 1-(1-(3-d3-methoxy-4-aminophenyl)piperidin-4-yl)-4-methylpiperazine (60 mg, 0.2 mmol Concentrated hydrochloric acid (two drops), the reaction mixture was heated to 100 ° C under N ₂ atmosphere, and stirred to react overnight. After cooling to room temperature, a saturated aqueous solution of sodium hydrogencarbonate (10 mL), EtOAc (EtOAc) %. LC-MS (APCI): m / z = 587.2 (M + 1) +; 1 H NMR (300MHz, DMSO-d 6) (δ / ppm) 11.18 (s, 1H), 8.51-8.47 (m, 1H) , 0.08-8.07 (m, 2H), 7.57-7.50 (m, 1H), 7.40-7.32 (m, 2H), 7.12-7.07 (m, 1H), 6.62 (d, J = 2.4 Hz, 1H), 6.47 (dd, J=9.0 Hz, 2.4 Hz, 1H), 3.77-3.72 (m, 2H), 2.82-2.61 (m, 11H), 2.48-2.34 (m, 3H), 1.91-1.85 (m, 2H), 1.79 (s, 3H), 1.74 (s, 3H), 1.59-1.52 (m, 2H).

Example 2

Preparation of 5-chloro-N ⁴ -[2-(dimethylphosphoryl)phenyl]-N ² -{2-methoxy-4-[4-(4-d3-methylpiperazine) according to the following synthetic route -1-yl)-piperidin-1-yl]phenyl}pyrimidine-2,4-diamine (compound 15):

(1) Preparation of Compound 10: N,N-dimethylformamide (10 mL) was added to a 100 mL one-neck flask, and 4-fluoro-2-methoxynitrobenzene (2 g, 11.8 mmol) and piperidine were sequentially added with stirring. 4-ketohydrochloride (2.23 g, 16.5 mmol), anhydrous potassium carbonate (4.88 g, 35.4 mmol), and the mixture was warmed to 80 ° C and allowed to react overnight under N ₂ atmosphere. After cooling to room temperature, poured into ice water (80 mL), EtOAc (EtOAc m. LC-MS (APCI): m / z = 251.2 (M + 1) +; 1 H NMR (300MHz, DMSO-d6) (δ / ppm) 7.93 (d, J = 9.6Hz, 1H), 6.62 (dd, J = 9.6 Hz, 2.4 Hz, 1H), 6.53 (d, J = 2.4 Hz, 1H), 3.94 (s, 3H), 3.84 (t, J = 6.3 Hz, 4H), 2.50 (t, J = 6.3 Hz) , 4H).

(2) Preparation of Compound 11: 20 mL of toluene was placed in a 100 mL one-necked flask, and 1-(3-methoxy-4-nitrophenyl)piperidin-4-one (0.53 g, 2.1 mmol) was added in that Triethylamine (0.8 mL), N-Boc piperazine (0.85 g, 4.5 mmol), stirred for 30 min under N ₂ atmosphere, sodium borohydride (0.4 g, 1.92 mmol) was added in one portion, stirred for 30 min, three times again 1.2 g of sodium borohydride hydride was added, and the reaction was completed. Add a saturated aqueous solution of sodium hydrogencarbonate (30 mL), EtOAc (EtOAc m. The solid was 0.58 g, and the yield was 65.7%. LC-MS (APCI): m / z = 421.2 (M + 1) +; 1 H NMR (300MHz, CDCl 3) (δ / ppm) 8.01 (d, J = 9.3Hz, 1H), 6.43 (dd, J = 9.6 Hz, 2.7 Hz, 1H), 6.31 (d, J = 2.4 Hz, 1H), 3.94 (s, 3H), 3.03-2.94 (m, 2H), 2.65-2.62 (m, 4H), 2.54-2.46 (m, 5H), 2.32 (s, 3H), 2.01-1.96 (m, 2H), 1.65-1.60 (m, 2H), 1.51 (s, 9H).

(3) Preparation of Compound 12: 20 mL of dichloromethane was added to a 100 mL single-necked flask, and 4-(1-(3-methoxy-4-nitrophenyl)piperidin-4-yl)piperazine was sequentially added with stirring. Base 1-tert-butyl ester (0.58g, 1.4mmol), trifluoroacetic acid (2mL), the reaction was stirred at room temperature for 1h under N ₂ atmosphere, the reaction solution was concentrated to dryness, and saturated aqueous sodium hydrogencarbonate (10 mL) was added. chloride extraction (20mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated to give a yellow solid 0.45g, yield 100%, LC-MS (APCI ): m / z = 321.2 (m + 1) +.

(4) Preparation of compound 13: 5 mL of acetonitrile was added to a 25 mL single-necked flask, and 4-(1-(3-methoxy-4-nitrophenyl)piperidin-4-yl)piperazine (0.32) was added in sequence with stirring. g, 1 mmol), triethylamine (0.12 g, 1.2 mmol) was added, and the mixture was cooled in ice-water-cooled. The product was passed through a silica gel column to obtain a yellow solid (0.15 g, yield: 44.5%). LC-MS (APCI): m / z = 338.2 (M + 1) +; 1 H NMR (300MHz, CDCl 3) (δ / ppm) 8.01 (d, J = 9.3Hz, 1H), 6.43 (dd, J = 9.6 Hz, 2.7 Hz, 1H), 6.31 (d, J = 2.4 Hz, 1H), 3.98-3.94 (m, 2H), 3.92 (s, 3H), 3.03-2.94 (m, 2H), 2.65-2.62 (m, 4H), 2.54-2.46 (m, 5H), 2.01-1.96 (m, 2H), 1.65-1.60 (m, 2H).

(5) Preparation of compound 14, which is prepared in the same manner as the preparation method of compound 5, except that 1-(1-(3-methoxy-4-nitrophenyl)piperidin-4-yl)-4-d3-methylpiperazine in place of 1-(1-(3-d3-methoxy-) 4-Nitrophenyl)piperidin-4-yl)-4-methylpiperazine.

(6) Preparation of 5-chloro-N ⁴ -[2-(dimethylphosphinyl)phenyl]-N ² -{2-methoxy-4-[4-(4-d3-methylpiperazine) 1-yl)-piperidin-1-yl]phenyl}pyrimidine-2,4-diamine (compound 14), which is prepared in the same manner as the preparation method of compound 9, except that 1-(1-) is used. (3-methoxy-4-aminophenyl)piperidin-4-yl)-4-d3-methylpiperazine in place of 1-(1-(3-d3-methoxy-4-aminobenzene) Basepiperidin-4-yl)-4-methylpiperazine. LC-MS (APCI): m / z = 587.2 (M + 1) +; 1 H NMR (300MHz, DMSO-d6) (δ / ppm) 11.18 (s, 1H), 8.51-8.47 (m, 1H), 0.08-8.07(m,2H), 7.57-7.50(m,1H), 7.40-7.32(m,2H),7.12-7.07(m,1H),6.62(d,J=2.4Hz,1H),6.47( Dd, J=9.0 Hz, 2.4 Hz, 1H), 3.76-3.71 (m, 6H), 2.82-2.61 (m, 11H), 2.48-2.34 (m, 3H), 1.91-1.85 (m, 2H), 1.79 (s, 3H), 1.74 (s, 3H), 1.59-1.52 (m, 2H).

Example 3

Preparation of 5-chloro-N ⁴ -[2-(dimethylphosphoryl)phenyl]-N ² -{2-methoxy-4-[4-(4-methylpiperazine-1) according to the following synthetic route -yl)-4-d-piperidin-1-yl]phenyl}pyrimidine-2,4-diamine (compound 20):

(1) Preparation of compound 16: 10 mL of deuterated methanol was added to a 50 mL one-necked flask, and 1-(3-methoxy-4-nitrophenyl)piperidin-4-one (0.25 g, 1 mmol) was added with stirring in an ice water bath. After completely dissolving, slowly add sodium borohydride (42 mg, 1 mmol), stir the reaction in an ice water bath under N ₂ atmosphere for 5 min, add heavy water (2 mL) to quench the reaction, and stir at room temperature for 30 min, then add water (30 mL) and Ethyl acetate (30 mL), EtOAc (EtOAc m. Dry over anhydrous sodium sulfate, filtered and concentrated to give 0.25 g of a yellow solid. LC-MS (ESI): m / z = 254.2 (M + 1) +; 1 H NMR (300MHz, DMSO-d 6) (δ / ppm) 7.88 (d, J = 9.3Hz, 1H), 6.58 (dd , J=9.3Hz, 2.4Hz, 1H), 6.49(d, J=2.4Hz, 1H), 4.75(s,1H), 3.90(s,3H),3.83-3.77(m,2H),3.23-3.14 (s, 2H), 1.84-1.76 (m, 2H), 1.45-1.37 (m, 2H).

(2) Preparation of Compound 17: Dichloromethane (15 mL) was added to a 50 mL single-necked flask, and 1-(3-methyl-4-nitrophenyl)-4-d-piperidin-4-ol was added in an ice water bath. (0.25 g, 1 mmol), triethylamine (0.18 g, 1.8 mmol) was added under stirring, and methylsulfonyl chloride (0.17 g, 1.5 mmol) was slowly added dropwise thereto, and the reaction was stirred at room temperature under N ₂ atmosphere for 1 h. Add water (20 mL), shake the organic layer, extract the aqueous layer with dichloromethane (20 mL x 2), and combine the organic layer with 0.5M HCl aqueous solution (20 mL x 1), saturated sodium bicarbonate (15 mL x) 1) Saturated brine (15 mL x 1), dried over anhydrous sodium sulfate, filtered and evaporated to dryness

(3) Preparation of Compound 18: DMF (3 mL) was added to a 25 mL single-necked flask, and 1-(3-methyl-4-nitrophenyl)-4-d-piperidine-4-methylsulfonate was added sequentially with stirring. The acid ester (0.3 g, 0.9 mmol), 1-methylpiperazine (0.36 g, 3.6 mmol), anhydrous potassium carbonate (0.62 g, 4.5 mmol), the mixture was warmed to 100 ° C, and stirred overnight under N ₂ atmosphere. . After cooling to room temperature, water (30 mL) and ethyl acetate (30 mL) were added, and the organic layer was separated, ethyl acetate was evaporated, ethyl acetate (20 <RTIgt; The sodium was dried, filtered, and concentrated. LC-MS (APCI): m / z = 339.2 (M + 1) +; 1 H NMR (300MHz, CDCl 3) (δ / ppm) 8.01 (d, J = 9.3Hz, 1H), 6.43 (dd, J = 9.6 Hz, 2.7 Hz, 1H), 6.31 (d, J = 2.4 Hz, 1H), 3.98-3.94 (m, 2H), 3.03-2.94 (m, 2H), 2.65-2.62 (m, 4H), 2.54 - 2.46 (m, 5H), 2.32 (s, 3H), 2.01-1.96 (m, 2H), 1.65-1.60 (m, 2H).

(4) Preparation of 1-[1-(3-methoxy-4-aminophenyl)-4-d-piperidin-4-yl]-4-methylpiperazine (Compound 19), a preparation method thereof Consistent with the preparation of Compound 5, except that 1-[1-(3-methoxy-4-nitrophenyl)-4-d-piperidin-4-yl]-4-methylper The azine is substituted for 1-[1-(3-d3-methoxy-4-nitrophenyl)piperidin-4-yl]-4-methylpiperazine.

(5) Preparation of 5-chloro-N ⁴ -[2-(dimethylphosphoryl)phenyl]-N ² -{2-methoxy-4-[4-(4-methylpiperazin-1- 4-)-piperidin-1-yl]phenyl}pyrimidine-2,4-diamine (compound 20), which is prepared in the same manner as the preparation method of compound 9, except that 1-[1 -(3-methoxy-4-aminophenyl)-4-d-piperidin-4-yl]-4-methylpiperazine in place of 1-[1-(3-d3-methoxy-4 -Aminophenyl)piperidin-4-yl]-4-methylpiperazine. LC-MS (APCI): m / z = 587.2 (M + 1) +; 1 H NMR (300MHz, DMSO-d6) δ (ppm): 11.18 (s, 1H), 8.51-8.47 (m, 1H), 0.08-8.07(m,2H), 7.57-7.50(m,1H), 7.40-7.32(m,2H),7.12-7.07(m,1H),6.62(d,J=2.4Hz,1H),6.47( Dd, J=9.0 Hz, 2.4 Hz, 1H), 3.76-3.71 (m, 6H), 2.82-2.61 (m, 10H), 2.48-2.34 (m, 3H), 1.91-1.85 (m, 2H), 1.79 (s, 3H), 1.74 (s, 3H), 1.59-1.52 (m, 2H).

Example 4

Preparation of 5-chloro -N ⁴ - [2- (dimethylphosphoryl) phenyl] -N ² - {2- methoxy-4- [4- (4-methyl -3,3,5,5 -d4-piperazin-1-yl)-piperidin-1-yl]phenyl}pyrimidine-2,4-diamine (Compound 21):

A method similar to that described in Example 2 was carried out except that 4-methyl-3,3,5,5-d4-piperazine was used instead of N-methylpiperazine to prepare the target compound.

Example 5

Preparation of 5-chloro-N ⁴ -[2-(dimethylphosphoryl)phenyl]-N ² -{2-methoxy-4-[4-(4-methyl 2,2,3,3, 5,5,6,6-d8-piperazin-1-yl)piperidin-1-yl]phenyl}pyrimidine-2,4-diamine (Compound 22):

A method similar to that described in Example 2, except that 4-methyl-2,2,3,3,5,5,6,6-d8-piperazine was used instead of N-methylpiperazine to obtain a target. Compound. LC-MS (APCI): m / z = 592.4 (M + 1) +; 1 H NMR (400MHz, CD 3 OD) (δ / ppm) 8.35 (dd, J = 8.4Hz, 4.4Hz, 1H), 8.04 (s, 1H), 7.69 (d, J = 8.8 Hz, 1H), 7.65-7.59 (m, 1H), 7.53 (t, J = 8 Hz, 1H), 7.29-7.25 (m, 1H), 6.67 (d , J = 2.4 Hz, 1H), 6.46 (dd, J = 8.8 Hz, 2.4 Hz, 1H), 3.86 (s, 3H), 3.71 (d, J = 12.8 Hz, 2H), 2.76-2.70 (m, 2H) ), 2.61-2.56 (m, 4H), 2.04 (d, J = 12.8 Hz, 2H), 1.87 (s, 3H), 1.83 (s, 3H), 1.76-1.65 (m, 2H).

Example 6

Preparation of 5-chloro-N ⁴ -[2-(dimethylphosphoryl)phenyl]-N ² -{2-d3-methoxy-4-[4-(4-d3-methyl) according to the following synthetic route Piperazin-1-yl)piperidin-1-yl]phenyl}pyrimidine-2,4-diamine (Compound 25):

(1) Preparation of Compound 23: 5 mL of acetonitrile was placed in a 25 mL single-necked flask, and 4-(1-(3-methoxy-4-nitrophenyl)piperidin-4-yl)piperazine (0.32) was added in sequence with stirring. g, 1 mmol), triethylamine (0.12 g, 1.2 mmol) was added, and the mixture was cooled in ice-water-cooled. The product was passed through a silica gel column to obtain a yellow solid (0.15 g, yield: 44.5%). LC-MS (APCI): m / z = 340.2 (M + 1) +.

(2) Preparation of Compound 24, which is produced in the same manner as in the preparation of Compound 5, except that 1-[1-(3-d3-methoxy-4-nitrophenyl)piperidin-4-yl is used. ]-4-d3-methylpiperazine in place of 1-[1-(3-d3-methoxy-4-nitrophenyl)piperidin-4-yl]-4-methylpiperazine.

(3) Preparation of Compound 25 in the same manner as in the preparation of Compound 9, except that 1-[1-(3-d3-methoxy-4-aminophenyl)piperidin-4-yl was used. ]-4-d3-methylpiperazine in place of 1-[1-(3-d3-methoxy-4-aminophenyl)piperidin-4-yl]-4-methylpiperazine. LC-MS (APCI): m / z = 587.2 (M + 1) +; 1 H NMR (300MHz, DMSO-d6) (δ / ppm) 11.18 (s, 1H), 8.51-8.47 (m, 1H), 0.08-8.07(m,2H), 7.57-7.50(m,1H), 7.40-7.32(m,2H),7.12-7.07(m,1H),6.62(d,J=2.4Hz,1H),6.47( Dd, J=9.0 Hz, 2.4 Hz, 1H), 3.76-3.71 (m, 6H), 2.82-2.61 (m, 11H), 2.48-2.34 (m, 3H), 1.91-1.85 (m, 2H), 1.79 (s, 3H), 1.74 (s, 3H), 1.59-1.52 (m, 2H).

Example 7

According to the following Scheme Preparation of 5-chloro -N ⁴ - [2- (dimethylphosphoryl) phenyl] -N ² - {2- methoxy -4- [4- (4-d3- methyl-2 , 2,3,3,5,5,6,6-d8-piperazin-1-yl)piperidin-1-yl]phenyl}pyrimidine-2,4-diamine (Compound 29):

(1) Preparation of Compound 27: Compound 26 (214 mg, 652 μmol), a heavy aqueous solution of deuterated formaldehyde (313 mg, 1.95 mmol, 20% / D ₂ O), and CH ₃ COOD (1 drop) were stirred at room temperature for 10 minutes. After adding sodium cyanoborohydride (129 mg, 1.95 mmol), stirring was continued for 30 minutes, then neutralized with triethylamine, concentrated, and purified by column chromatography to yield 175 mg of a yellow solid. LC-MS (APCI): m / z = 346.4 (M + 1) +.

(2) Preparation of Compound 28 in the same manner as in the preparation of Compound 5, except that 1-[1-(3-methoxy-4-nitrophenyl)piperidin-4-yl]- 4-d3-methyl-2,2,3,3,5,5,6,6-d8-piperazine in place of 1-[1-(3-d3-methoxy-4-nitrophenyl)per Pyridin-4-yl]-4-methylpiperazine.

(3) Preparation of Compound 29 in the same manner as in the preparation of Compound 9, except that 1-[1-(methoxy-4-aminophenyl)piperidin-4-yl]-4- D3-methyl-2,2,3,3,5,5,6,6-d8-piperazine in place of 1-[1-(3-d3-methoxy-4-aminophenyl)piperidine- 4-yl]-4-methylpiperazine. LC-MS (APCI): m / z = 595.4 (M + 1) +; 1 H NMR (300MHz, CDCl 3) (δ / ppm) 10.80 (s, 1H), 8.63 (dd, J = 4.8Hz, 2.4 Hz, 1H), 8.09-8.07 (m, 2H), 7.50 (t, J = 4.5 Hz, 1H), 7.30-7.25 (m, 2H), 7.14-7.10 (m, 1H), 6.55 (d, J = 1.5 Hz, 1H), 6.49 (dd, J = 5.4 Hz, 1.5 Hz, 1H), 3.87 (s, 3H), 3.66 (d, J = 7.5 Hz, 2H), 2.73 - 2.68 (m, 2H), 2.40 - 2.36 (m, 1H), 1.95 (d, J = 7.5 Hz, 2H), 1.85 (s, 3H), 1.82 (s, 3H), 1.76-1.68 (m, 2H).

Example 8

According to the following Scheme Preparation of 5-chloro -N ⁴ - [2- (dimethylphosphoryl) phenyl] -N ² - ^{{2-d3--methoxy-4-} [4- (4-methyl-2 , 2,3,3,5,5,6,6-d8-piperazin-1-yl)piperidin-1-yl]phenyl}pyrimidine-2,4-diamine (Compound 32):

(1) Compound 30 was prepared in the same manner as in the preparation of Compound 10 except that d3-5-fluoro-2-nitroanisole was used instead of 5-fluoro-2-nitroanisole. LC-MS (APCI): m / z = 254.5 (M + 1) +.

(2) Preparation of compound 31: tetraisopropyl titanate (Ti(Oi-Pr) ₄ , 5 mL) was added to compound 30 (900 mg, 3.6 mmol) and ₂ , ₂ , ₃ , ₃ , 5, 5, 6, A solution of 6-d8-piperazine (474 mg, 5.03 mmol) was stirred at room temperature overnight, then 10 mL of ethanol was added, and sodium nitricarb borohydride (678 mg, 10.79 mmol) was added and the mixture was stirred at room temperature for 3 hours. The water (10 mL) in which 5 g of celite was dissolved was poured, and stirring was continued for 30 minutes. The solvent was removed, and then purified by column chromatography to afford product (yield: 25.4%). LC-MS (APCI): m / z = 332.5 (M + 1) +.

(3) Compound 32 was prepared in the same manner as in the preparation of Compound 29 except that Compound 31 was used instead of Compound 26, and formaldehyde was substituted for deuterated formaldehyde, and sodium nitrile borohydride was used instead of sodium deuterated borohydride. The target product was finally obtained as a white solid, 40 mg in total, yield 53.0%. LC-MS (APCI): m / z = 595.5 (M + 1) +; 1 H NMR (300MHz, CDCl 3) (δ / ppm) 10.80 (s, 1H), 8.62 (dd, J = 8.1Hz, 4.5 Hz, 1H), 8.11-8.08 (m, 2H), 7.50 (t, J = 7.8 Hz, 1H), 7.32-7.25 (m, 2H), 7.16-7.11 (m, 1H), 6.54 (d, J = 1.6 Hz, 1H), 6.48 (dd, J = 8.4 Hz, 2.4 Hz, 1H), 3.66 (d, J = 12 Hz, 2H), 2.74 - 2.67 (m, 2H), 2.61-2.55 (m, 1H), 2.48 (s, 3H), 2.02 (d, J = 12.3 Hz, 2H), 1.84 (s, 3H), 1.81 (s, 3H), 1.79-1.73 (m, 2H).

Example 9

Preparation of 5-chloro-N ⁴ -[2-(dimethylphosphoryl)phenyl]-N ² -{2-d3-methoxy-4-[4-(4-d2-methyl-2,2 ,3,3,5,5,6,6-d8-piperazin-1-yl)piperidin-1-yl]phenyl}pyrimidine-2,4-diamine (Compound 33):

Similar to the method described in Example 7, except that Compound 31 was used instead of Compound 26, and sodium nitrile borohydride was substituted for sodium deuterated borohydride. The target product was finally obtained as a yellow solid, a total of 70 mg, yield 27.2%. LC-MS (APCI): m / z = 597.4 (M + 1) +; 1 H NMR (300MHz, CDCl 3) (δ / ppm) 10.82 (s, 1H), 8.62 (dd, J = 8.4Hz, 4.5 Hz, 1H), 8.13 - 8.09 (m, 2H), 7.50 (t, J = 7.5 Hz, 1H), 7.33 - 7.26 (m, 2H), 7.16 - 7.10 (m, 1H), 6.54 (d, J = 2.1 Hz, 1H), 6.48 (dd, J = 9.0 Hz, 2.4 Hz, 1H), 3.66 (d, J = 12.6 Hz, 2H), 2.76-2.59 (m, 3H), 2.56 (s, 1H), 2.08 -2.00 (m, 2H), 1.86 (s, 3H), 1.81 (s, 3H), 1.79-1.72 (m, 2H).

Example 10

Preparation of 5-chloro-N ⁴ -[2-(dimethylphosphoryl)phenyl]-N ² -{2-d3-methoxy-4-[4-(4-d3-methyl-2,2 ,3,3,5,5,6,6-d8-piperazin-1-yl)piperidin-1-yl]phenyl}pyrimidine-2,4-diamine (compound 34):

Similar to the method described in Example 7, except that Compound 31 was substituted for Compound 26. The target product was finally obtained as a white solid, a total of 110 mg, yield 36.7%. LC-MS (APCI): m / z = 598.4 (M + 1) +; 1 H NMR (300MHz, CDCl 3) (δ / ppm) 8.35 (dd, J = 8.4Hz, 4.4Hz, 1H), 8.04 ( s, 1H), 7.68 (d, J = 8.4 Hz, 1H), 7.65-7.59 (m, 1H), 7.52 (t, J = 8 Hz, 1H), 7.29-7.25 (m, 1H), 6.67 (d, J=6.8 Hz, 1H), 6.46 (dd, J=8.8 Hz, 2.4 Hz, 1H), 3.71 (d, J = 12.4 Hz, 2H), 2.76-2.70 (m, 2H), 2.64-2.58 (m, 1H), 2.04 (d, J = 12.4 Hz, 2H), 1.87 (s, 3H), 1.83 (s, 3H), 1.76-1.66 (m, 2H).

Example 11

Biological evaluation of compounds

The compounds of the invention were evaluated in a number of tests to determine their biological activity.

(1) Evaluation of kinase inhibition

Compound Formulation: Test compounds were dissolved in DMSO to make a 20 mM stock solution. Compounds were diluted to 0.1 mM in DMSO (100 times the final concentration of the dilution) before use and diluted in 3 folds at 11 concentrations. Dilute to 4 times the final concentration of the dilution solution with the buffer.

Kinase assay: After the buffer was prepared, the enzyme was mixed with different concentrations of the compound prepared by pre-diluting, and allowed to stand at room temperature for 30 minutes at each concentration. The corresponding substrate and ATP were added and reacted at room temperature for 60 minutes (in which a negative positive control was set). After the reaction was completed, the antibody was added for detection. After incubation at room temperature for 60 minutes, Evnvision was detected and data was collected. Data analysis and mapping according to XLfit5 software. Crizotinib was used as a control.

Table 1 Comparison of the kinase inhibition effects of Examples 1 to 10 and the control of crizotinib

As shown in Table 1, the compound of the present invention exhibited excellent inhibitory activity against the ALK L1196M mutant (IC ₅₀ less than 20 nM) as compared with the existing ALK inhibitor crizotinib, indicating that the compound of the present invention is highly potent against ALK. Inhibition ability.

(2) Cytotoxicity experiment

The inhibitory effects of the compounds of Examples 1 to 10 on tumor cells were examined by the tetrazolium salt (MTS) method, and crizotinib was used as a control. The experimental results are shown in Table 2. Compared to the existing ALK inhibitor crizotinib, the compounds of the present invention all exhibited excellent anticancer activity against the growth of the ALK mutant L1196M cancer cells.

Table 2 Comparison of cytotoxicity experiments of Examples 1 to 10 and the control of crizotinib

(3) Metabolic stability evaluation

Microsomal experiments: human liver microsomes: 0.5 mg/mL, Xenotech; rat liver microsomes: 0.5 mg/mL, Xenotech; coenzyme (NADPH/NADH): 1 mM, Sigma Life Science; magnesium chloride: 5 mM, 100 mM phosphate buffer Agent (pH 7.4).

Preparation of the stock solution: A certain amount of the compound of the example was accurately weighed and dissolved to 5 mM with DMSO.

Preparation of phosphate buffer (100 mM, pH 7.4): Mix 150 mL of pre-formed 0.5 M potassium dihydrogen phosphate and 700 mL of 0.5 M potassium dihydrogen phosphate solution, and adjust the mixture with 0.5 M potassium dihydrogen phosphate solution. The pH was adjusted to 7.4, diluted 5 times with ultrapure water before use, and magnesium chloride was added to obtain a phosphate buffer (100 mM) containing 100 mM potassium phosphate, 3.3 mM magnesium chloride, and a pH of 7.4.

A solution of NADPH regeneration system (containing 6.5 mM NADP, 16.5 mM G-6-P, 3 U/mL G-6-P D, 3.3 mM magnesium chloride) was prepared and placed on wet ice before use.

Formulation stop solution: acetonitrile solution containing 50 ng/mL propranolol hydrochloride and 200 ng/mL tolbutamide (internal standard). Take 25057.5 μL of phosphate buffer (pH 7.4) into a 50 mL centrifuge tube, add 812.5 μL of human liver microsomes, and mix to obtain a liver microsome dilution with a protein concentration of 0.625 mg/mL. 25057.5 μL of phosphate buffer (pH 7.4) was taken into a 50 mL centrifuge tube, and 812.5 μL of SD rat liver microsomes were added and mixed to obtain a liver microsome dilution having a protein concentration of 0.625 mg/mL.

Incubation of the sample: The stock solution of the corresponding compound was diluted to 0.25 mM with an aqueous solution containing 70% acetonitrile. As a working fluid, spare. 398 μL of human liver microsomes or rat liver microsome dilutions were added to 96-well incubation plates (N=2), and 2 μL of 0.25 mM working solution was added and mixed.

Determination of metabolic stability: 300 μL of pre-cooled stop solution was added to each well of a 96-well deep well plate and placed on ice as a stop plate. The 96-well incubation plate and the NADPH regeneration system were placed in a 37 ° C water bath, shaken at 100 rpm, and pre-incubated for 5 min. 80 μL of the incubation solution was taken from each well of the incubation plate and added to the stopper plate, and mixed, and 20 μL of the NADPH regeneration system solution was added as a sample of 0 min. Then, 80 μL of the NADPH regeneration system solution was added to each well of the incubation plate to start the reaction and start timing. The corresponding compound had a reaction concentration of 1 μM and a protein concentration of 0.5 mg/mL. 100 μL of the reaction solution was taken at 10, 30, and 90 min, respectively, and added to the stopper, and the reaction was terminated by vortexing for 3 min. The plate was centrifuged at 5000 x g for 10 min at 4 °C. 100 μL of the supernatant was taken into a 96-well plate to which 100 μL of distilled water was previously added, mixed, and sample analysis was performed by LC-MS/MS.

Data analysis: The peak area of the corresponding compound and the internal standard was detected by LC-MS/MS system, and the ratio of the peak area of the compound to the internal standard was calculated. The slope is measured by the natural logarithm of the percentage of the remaining amount of the compound versus time, and t _1/2 and CL _{int are} calculated according to the following formula, where V/M is equal to 1/protein concentration.

The metabolic stability of human and rat liver microsomes was evaluated by comparing the compounds of the present invention and their compounds without deuteration. The half-life and liver intrinsic clearance (Clint) as indicators of metabolic stability are shown in Table 3. The undeuterated compound AP26113 was used as a control sample in Table 3. As shown in Table 3, the compounds of the present invention can significantly improve metabolic stability by comparison with the undeuterated compound AP26113, and are thus more suitable for the preparation of metastatic ALK-positive non-small cells for the treatment of crizotinib tolerance. Drugs for lung cancer.

Table 3 Comparison of metabolic stability of Examples 1 to 10 and AP26113 control

(4) Pharmacokinetic evaluation in rats

Six male Sprague-Dawley rats, 7-8 weeks old, weighing 210 g, divided into 2 groups, 3 in each group, intravenously or orally administered a single dose of compound (3 mg/kg intravenously, 10 mg/kg orally). Its pharmacokinetic differences. Rats were fed a standard diet and given water. Fasting began 16 hours before the test. The drug was dissolved with PEG400 and dimethyl sulfoxide. Blood was collected from the eyelids at a time point of 0.083 hours, 0.25 hours, 0.5 hours, 1 hour, 2 hours, 4 hours, 6 hours, 8 hours, 12 hours, and 24 hours after administration. Rats were briefly anesthetized after inhalation of ether, and 300 μL of blood samples were collected from the eyelids in test tubes. There was 30 μL of 1% heparin salt solution in the test tube. The tubes were dried overnight at 60 ° C before use. After the blood sample collection was completed, the rats were anesthetized with ether and sacrificed. Immediately after the blood sample is collected, gently invert the tube at least 5 times to ensure adequate mixing and place on ice. Blood samples were centrifuged at 5000 rpm for 5 minutes at 4 ° C to separate plasma from red blood cells. Pipette 100 μL of plasma into a clean plastic centrifuge tube to indicate the name and time point of the compound. Plasma was stored at -80 °C prior to analysis. The concentration of the compound of the invention in plasma was determined by LC-MS/MS. Pharmacokinetic parameters were calculated based on the plasma concentration of each animal at different time points.

The experimental results are shown in Table 4 below. Compared with the control compound AP26113, the oral utilization rate (F) of the compound 15 of Example 2 of the present invention is comparable to that of the control compound AP26113, but its half-life is prolonged and the metabolic stability is remarkably improved; The oral availability of Compound 9 was significantly increased (up 20%), indicating better pharmacokinetics in animals.

Table 4 Rat pharmacokinetic experiments

It is to be understood that the examples are merely illustrative of the invention and are not intended to limit the scope of the invention, and the experimental methods in which the specific conditions are not indicated in the examples, usually in accordance with conventional conditions or in accordance with the conditions suggested by the manufacturer. Parts and percentages are parts by weight and percentage by weight unless otherwise stated. The above is a further detailed description of the present invention in connection with the specific preferred embodiments, and the specific embodiments of the present invention are not limited to the description. It will be apparent to those skilled in the art that the present invention may be made without departing from the spirit and scope of the invention.

Claims

A diaminopyrimidine compound characterized by a diaminopyrimidine compound represented by formula (I), or a crystalline form thereof, a pharmaceutically acceptable salt, a hydrate or a solvent compound,

Wherein R 1a , R 1b , R 1c , R 2a , R 2b , R 3a , R 3b , R 4a , R 4b , R 5a , R 5b , R 6 , R 7a , R 7b , R 8a , R 8b , R 9a , R 9b , R 10a , R 10b , R 11 , R 12 , R 13 , R 14a , R 14b , R 14c , R 15 , R 16 , R 17a , R 17b , R 17c , R 18a , R 18b And R 18c , R 19 , R 20 , R 21 and R 22 are each independently hydrogen, deuterium, halogen or trifluoromethyl;

R 16 is hydrogen, deuterium, halogen, cyano, unsubstituted C 1 -C 6 alkyl or C 1 -C 6 alkoxy, one or more deuterated or fully deuterated C 1 -C 6 An alkyl or C 1 -C 6 alkoxy group, or one or more halogen-substituted or perhalogen-substituted C 1 -C 6 alkyl or C 1 -C 6 alkoxy;

Additional conditions are R 1a , R 1b , R 1c , R 2a , R 2b , R 3a , R 3b , R 4a , R 4b , R 5a , R 5b , R 6 , R 7a , R 7b , R 8a , R 8b And R 9a , R 9b , R 10a , R 10b , R 11 , R 12 , R 13 , R 14a , R 14b , R 14c , R 15 , R 16 , R 17a , R 17b , R 17c , R 18a , R At least one of 18b , R 18c , R 19 , R 20 , R 21 and R 22 is deuterated or deuterated.
The diaminopyrimidine compound according to claim 1, wherein R 1a , R 1b and R 1c are hydrazine.
The diaminopyrimidine compound according to claim 1, wherein: R 7a, R 7b, R 8a, R 8b, R 9a, R 9b, R 10a and R 10b are each independently hydrogen or deuterium.
The diaminopyrimidine compound according to claim 1, wherein R 14a , R 14b and R 14c are hydrazine.
The diaminopyrimidine compound according to claim 1, wherein the compound is selected from the group consisting of the following compounds or a pharmaceutically acceptable salt thereof:
A method for producing a pharmaceutical composition of a diaminopyrimidine compound according to any one of claims 1 to 5, wherein a pharmaceutically acceptable carrier and the compound described in the first aspect of the invention, or The crystalline form, pharmaceutically acceptable salt, prodrug, stereoisomer, isotopic variant hydrate or solvate is combined to form a pharmaceutical composition.
A pharmaceutical composition comprising a pharmaceutically acceptable carrier and the diaminopyrimidine compound according to any one of claims 1 to 5, or a crystalline form thereof, a pharmaceutically acceptable salt, or a hydrate thereof Or a pharmaceutical composition of a solvate, stereoisomer, prodrug or isotopic variation.
The pharmaceutical composition according to claim 7, which further comprises other therapeutic agents, which are cancer, cardiovascular disease, inflammation, infection, immune disease, cell proliferative disease, viral disease, A drug for metabolic diseases, or organ transplantation.
Use of a diaminopyrimidine compound according to any one of claims 1 to 5, or a crystalline form thereof, a pharmaceutically acceptable salt, a hydrate or a solvate thereof, for the preparation of an inhibitor of ALK kinase Pharmaceutical composition.