CN107382879B - Pyrimidine compound, EGFR inhibitor and application thereof - Google Patents
Pyrimidine compound, EGFR inhibitor and application thereof Download PDFInfo
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- C07D401/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a chain containing hetero atoms as chain links
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- C07D403/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings linked by a chain containing hetero atoms as chain links
Abstract
The invention discloses a pyrimidine compound, an EGFR inhibitor and application thereof. The pyrimidine compound comprises a compound shown as a formula I, or pharmaceutically acceptable salt, stereoisomer, solvate or prodrug thereof; the EGFR inhibitor comprises the pyrimidine compounds. The pyrimidine compounds of the invention can inhibit activation or resistance mutation of one or more EGFR; the double-mutation EGFR T790M/L858R enzyme can be inhibited from proliferating under nanomolar concentration, and the inhibition on wild type EGFR enzyme is relatively weak; the composition can be used for treating EGFR sensitive type mutant cancer, and is also suitable for the case of generating secondary drug resistance in the current EGFR-TKI treatment; meanwhile, the mutation selectivity of the compound greatly reduces the toxic and side effects caused by inhibiting the wild EGFR, and the compound is an ideal medicine for treating diseases caused by EGFR mutation.
Description
Technical Field
The invention belongs to the technical field of medicines, and particularly relates to a pyrimidine compound, an EGFR inhibitor and application thereof in preparing medicines for regulating the activity of EGFR tyrosine kinase or treating EGFR related diseases, in particular non-small cell lung cancer.
Background
The epidermal Growth Factor receptor egfr (epidermal Growth Factor receptor) is one of the transmembrane protein tyrosine kinases of the erbB receptor family. When bound to a growth factor ligand, such as Epidermal Growth Factor (EGF), the receptor may homodimerize with additional EGFR molecules or heterodimerize with another family member, such as erbB2(HER2), erbB3(HER3), or erbB4(HER 4). Homodimerization and/or heterodimerization of erbB receptors results in phosphorylation of key tyrosine residues in the intracellular domain and in stimulation of many intracellular signaling pathways involved in cell proliferation and survival. Dysregulation of erbB family signaling promotes proliferation, invasion, metastasis, angiogenesis, and tumor cell survival, and has been described in many human cancers (including lung, head and neck, and breast cancers, among others).
Therefore, many drugs that represent the erbB family as rational targets for the development of anticancer drugs, such as those targeting EGFR or erbB2, have now been widely used clinically, including gefitinib (IRESSA)TM) Erlotinib (TARCEVA)TM) And lapatinib (TYKERB)TM) And the like. A detailed discussion of erbB receptor signaling and its involvement in tumorigenesis is provided in New England Journal of Medicine (2008, stage 358, 1160-1174) and Biochemical and Biophysical Research Communications (2004, stage 319, 1-11).
Lung cancer is the most prevalent cancer worldwide, the first of all cancers to have chinese incidence, and the highest incidence and mortality, and approximately 30% of lung cancer patients in china have EGFR mutations, with L858R and exon19 deletion mutations accounting for more than about 90% of these patients being more sensitive to EGFR inhibitors. The first-generation EGFR inhibitors such as erlotinib and gefitinib which are on the market at present have better curative effect on the patients, can reduce the tumor of more than 60 percent of the patients and obviously prolong the progression-free survival period of the patients. However, most patients acquire resistance at 6-12 months, and this resistance pattern is a further mutation of EGFR, which reduces their sensitivity to first generation EGFR inhibitors. The most common of these mutations is the so-called "gatekeeper" mutation T790M (Science,2004, Vol.304, 1497-1500; New England Journal of Medicine 2004,350, 2129. sub.2139), which is replaced by L-methionine (M) which is originally present at this site, and the mutated EGF tyrosine kinase R no longer binds to gefitinib or erlotinib, so that the first generation EGFR inhibitor will no longer be effective, resulting in the current state of drug unavailability of such patients. It was found that 50% of patients who developed resistance to first generation EGFR inhibitors tested for the EGFR T790M mutation. The first generation EGFR inhibitors, such as gefitinib and erlotinib, were greater than 3 μ M in T790M mutant cell line H1975 and were essentially inactive.
The curative effect of the second generation irreversible pan-EGFR inhibitor (Afatinib BIBW2992) which is developed and marketed at present on EGFR mutant lung cancer patients is remarkably better than that of the first generation EGFR inhibitor. However, the second generation inhibitor also has strong wild type EGFR inhibitory activity, the inhibitory activity to the wild type EGFR is obviously higher than that to drug-resistant T790M mutation, the toxic and side effects such as rash of patients are serious, the curative effect of drug-resistant patients is poor, and only a small part of drug-resistant patients of the first generation EGFR inhibitor respond to the drugs.
In order to improve the inhibitory activity on mutations such as drug-resistant EGFR T790M and reduce the inhibitory activity on wild type EGFR, the development of a third-generation EGFR mutant selective inhibitor with higher activity, better selectivity and lower toxicity has important significance.
Disclosure of Invention
The invention aims to provide a pyrimidine compound which has good inhibitory activity on EGFR.
It is a second object of the present invention to provide an EGFR inhibitor.
The third object of the present invention is to provide a use of the above-mentioned EGFR inhibitor for the preparation of a medicament for modulating EGFR tyrosine kinase activity or treating EGFR-related diseases.
In order to achieve the above purpose, the technical scheme adopted by the invention is as follows:
a pyrimidine compound, comprising a compound of formula I, or a pharmaceutically acceptable salt, stereoisomer, solvate, or prodrug thereof:
wherein Ar is selected from phenyl or substituted phenyl or quinolyl or substituted quinolyl;
R1selected from hydrogen, halogen, trifluoromethyl or cyano;
R2selected from methoxy, difluoromethoxy, difluorodeuterated methoxy or trifluoromethoxy;
R3any one structure selected from the following:
X1、X2、X3each independently selected from hydrogen or halogen.
When Ar is substituted phenyl, the substituted phenyl is mono-substituted, di-substituted or tri-substituted phenyl, and the substituents are respectively and independently selected from halogen, cyano, nitro, ester group and C1-4Alkyl or cycloalkyl, C1-4Alkoxy or cycloalkoxy, C1-4Haloalkyl, C1-4Acyl radical, C1-6Alkylamino or cycloalkylamino. The quinolyl group is 6-quinolyl.
Preferably, in the pyrimidine compounds, the compound shown in formula I is selected from:
the pharmaceutically acceptable salt is an inorganic acid salt or an organic acid salt, and the inorganic acid salt is selected from hydrochloride, hydrobromide, hydroiodide, sulfate, bisulfate, nitrate, phosphate and acid phosphate; the organic acid salt is selected from formate, acetate, trifluoroacetate, propionate, pyruvate, glycolate, oxalate, malonate, fumarate, maleate, lactate, malate, citrate, tartrate, methanesulfonate, ethanesulfonate, benzenesulfonate, salicylate, picrate, glutamate, salicylate, ascorbate, camphorate, camphorsulfonate.
The pyrimidine compound is a compound shown in a formula I, or pharmaceutically acceptable salt, stereoisomer, solvate or prodrug thereof, and can inhibit one or more activation or resistance mutations of EGFR (epidermal growth factor receptor), such as an L858R activation mutant, an Exon19 deletion EGFR activation mutant and a T790M resistance mutant; the compound improves the inhibition activity on mutations such as drug-resistant EGFRT790M and the like, reduces the inhibition activity on wild type EGFR, and can be used for developing third-generation EGFR mutant selective inhibitors with higher activity, better selectivity and lower toxicity.
In vitro experiments show that the pyrimidine compound can inhibit the proliferation of EGFR T790M/L858R double mutant enzyme under nanomolar concentration, and the inhibition on wild type EGFR enzyme is relatively weak. Therefore, the compounds can be used for treating EGFR sensitive type mutation cancers and also are applicable to the cases generating secondary drug resistance in the current EGFR-TKI treatment; meanwhile, the mutation selectivity of the compound greatly reduces the toxic and side effects caused by inhibiting the wild EGFR, and the compound is an ideal medicine for treating diseases caused by EGFR mutation.
The solvate referred to in the present invention means a complex formed by the compound of the present invention and a solvent. They either react in a solvent or precipitate out of a solvent or crystallize out. For example, the complex formed with water is known as a hydrate; others include alcoholates, ketonates, and the like. The solvate of the invention comprises a compound shown in the formula I, and a solvate of salt and stereoisomer thereof.
The stereoisomers mentioned in the present invention mean that the compounds of formula I according to the invention may contain one or more chiral centers and exist in different optically active forms. When the compound contains one chiral center, the compound comprises enantiomers. The present invention includes both isomers and mixtures of isomers, such as racemic mixtures. Enantiomers can be resolved by methods known in the art, such as crystallization and chiral chromatography. When the compounds of formula I contain more than one chiral center, non-corresponding isomers may exist. Stereoisomers of the present invention include resolved optically pure specific isomers as well as mixtures of non-corresponding isomers. Diastereomers may be resolved by methods known in the art, such as crystallization and preparative chromatography.
Prodrugs as referred to herein are intended to include known amino and carboxyl protecting groups, and are intended to be hydrolyzed under physiological conditions or released via enzymatic reaction to give the parent compound. Specific prodrug preparation methods can be referred to in the art (Saulnier, M.G.; Frannesson, D.B.; Deshpande, M.S.; Hansel, S.B and Vysa, D.M.Bioorg.Med.ChemLett.1994,4, 1985-oz 1990; and Greenwald, R.B.; Choe, Y.H.; Conover, C.D.; Shum, K.; Wu D.; Royzen, M.J.Med.Chem.2000, 43, 475.).
Dissolving the intermediate A, the intermediate B and p-toluenesulfonic acid in an organic solvent, reacting at 50-100 ℃ in a protective atmosphere, adding dichloromethane and a saturated sodium carbonate aqueous solution after the reaction is finished, layering, taking an organic phase, removing the solvent, and separating and purifying to obtain the pyrimidine compound;
the structural formulas of the intermediate A and the intermediate B are respectively as follows:
wherein, Ar and R1、R2、R3、X1、X2、X3As defined in formula I.
The preparation method relates to the following reaction formula:
the molar ratio of the intermediate A to the intermediate B is 1-5: 1. The p-toluenesulfonic acid is added in the form of p-toluenesulfonic acid monohydrate; the molar ratio of the p-toluenesulfonic acid to the intermediate B is 0.5-2: 1. The organic solvent is 2-pentanol; the dosage of the organic solvent is as follows: 5mL of organic solvent was used for every 50mg of intermediate B. The protective atmosphere is nitrogen. The separation and purification is separation by column chromatography.
The intermediate A is prepared by the following method:
the method comprises the following steps: mixing the compound a1 with diisopropylethylamine and n-butanol to obtain a mixture, cooling the mixture to-20 ℃, adding the compound a2 for reaction, heating to room temperature, stirring overnight, removing the solvent of the reaction system, adding ethyl acetate and water into the residue, layering, taking an organic phase, removing the solvent, and separating and purifying to obtain the compound A-S-D-methyl ethyl-ethylamine;
alternatively, method 2: mixing the compound a1 with diisopropylethylamine, n-butanol and the compound a2 to obtain a mixture, heating the mixture to 100 ℃ for reacting overnight, removing a solvent of a reaction system, adding ethyl acetate and water into residues, layering, taking an organic phase, removing the solvent, separating and purifying to obtain the compound a 1;
the structural formulas of the compound a1 and the compound a2 are shown as follows:
wherein, Ar and R1As defined in formula (I).
The preparation method of the intermediate A relates to the following reaction formula:
wherein the molar ratio of the compound a1 to the compound a2 is 1: 0.5-2. The dosage of the diisopropylethylamine is as follows: 1-3 mL of diisopropylethylamine is added to every 4-9 mmol of the compound a 1. The dosage of the n-butanol is as follows: 20-30 mL of n-butanol is added for every 4-9 mmol of the compound a 1. In the preparation of the intermediate A, the separation and purification are performed by column chromatography. The developing solvent used for the column chromatography is a mixture of ethyl acetate and petroleum ether.
Preferably, the intermediate a is selected from the following compounds:
the intermediate B is prepared by the following method:
preparing compound b1 into phenol sodium salt, and mixing with methyl iodide or ethyl difluorobromoacetate or mixture of deuterium water and ethyl difluorobromoacetate (R)2Precursor of (b) undergo substitution reaction, then undergo reduction (hydrogenation), nitration, protection of amino group with di-tert-butyl dicarbonate, and then undergo substitution reaction with substituent R3The precursor (B) undergoes a substitution reaction, followed by reduction (hydrogenation), and then reacts with acrylic acid, halogen-substituted acrylic acid, or acid chloride to give an intermediate B.
The involved reaction formula is as follows:
in the above-mentioned process for producing intermediate B, the intermediate B can be obtained from any step up to the time when starting materials are available in the prior art.
Preferably, intermediate B is selected from the following compounds:
an EGFR inhibitor comprises the pyrimidine compound.
The EGFR inhibitor further comprises a pharmaceutically acceptable carrier.
The EGFR inhibitor can be the compound shown in the formula I, or pharmaceutically acceptable salt, stereoisomer, solvate or prodrug thereof, and can also be a pharmaceutical composition containing the compound. The pharmaceutical composition further comprises a pharmaceutically acceptable carrier.
The EGFR inhibitor of the present invention can be administered by combining the compound shown in formula I, or pharmaceutically acceptable salt, stereoisomer, solvate or prodrug thereof, with one or more pharmaceutically acceptable carriers to form a suitable dosage form. These dosage forms are suitable for oral, rectal, topical, oral, and other parenteral administration (e.g., subcutaneous, intramuscular, intravenous, etc.).
Where the EGFR inhibitor of the present invention is a pharmaceutical composition, the composition is formulated, dosed and administered in a manner consistent with medical practice specifications. The "effective amount" of a compound to be administered will depend on, among other factors, the particular condition being treated, the individual being treated, the cause of the condition, the target of the drug, and the mode of administration.
The application of the EGFR inhibitor in preparing medicines for regulating the activity of EGFR tyrosine kinase or treating EGFR related diseases.
The EGFR tyrosine kinase activity regulation or EGFR related disease treatment refers to cancer, diabetes, immune system diseases, neurodegenerative diseases or cardiovascular diseases.
An application of the EGFR inhibitor in preparing a medicament for treating non-small cell lung cancer. The EGFR inhibitor of the present invention is particularly useful for the preparation of a medicament for the treatment of cancer, such as non-small cell lung cancer.
The EGFR inhibitor can be used for preparing medicaments for regulating the activity of EGFR tyrosine kinase or treating EGFR related diseases, such as cancers, diabetes, immune system diseases, neurodegenerative diseases or cardiovascular diseases and the like, and is particularly suitable for treating non-small cell lung cancer caused by EGFR mutation, including sensitive mutation (such as L858R mutation or exon19 deletion) and drug-resistant mutation (such as EGFRT790M mutation).
The EGFR inhibitor of the present invention, which comprises a compound represented by formula I, or a pharmaceutically acceptable salt, stereoisomer, solvate or prodrug thereof, can be used as a monotherapy in anticancer therapy, or in addition to that, can be used in combination with conventional surgery or radiotherapy or chemotherapy or immunotherapy. The above therapies may be administered concurrently, simultaneously, sequentially, or separately with an EGFR inhibitor of the present invention.
The medicament for modulating the activity of EGFR tyrosine kinase or treating EGFR-related diseases may further comprise any one or more of the following drugs in addition to the EGFR inhibitor of the present invention: gefitinib, erlotinib, lapatinib, XL647, NVP-AEE-788, ARRY-334543, vandetanib, PF00299804, cetuximab, panitumumab, pertuzumab, zalutumumab, nimotuzumab, MDX-214, CDX-110, IMC-11F8, CNF2024, tanespimycin, apramycin, IPI-504, NVP-AUY 922.
Detailed Description
The present invention will be further described with reference to the following embodiments.
In specific embodiments, eq is the molar equivalent of the reactants.
In a specific embodiment, the synthesis of intermediate a and intermediate B used is as follows. In the synthesis reaction formula of the intermediate A and the intermediate B, Ar is selected from phenyl or substituted phenyl, the substituted phenyl is mono-substituted, di-substituted or tri-substituted phenyl, and the substituents are respectively and independently selected from halogen, cyano, nitro, ester group and C1-4Alkyl or cycloalkyl, C1-4Alkoxy or cycloalkoxy, C1-4Haloalkyl, C1-4Acyl radical, C1-6Alkylamino or cycloalkylamino; r1Selected from hydrogen, halogen, trifluoromethyl or cyano; r2Selected from methoxy, difluoromethoxy, difluorodeuterated methoxy or trifluoromethoxy; r3Any one structure selected from the following:
X1、X2、X3each independently selected from hydrogen or halogen.
The preparation method of the intermediate a is method a1, method a2 or method A3, as follows.
Method A1: adding 7.8mmol of compound a1-1 into a 100mL single-neck bottle, and adding 3mL of diisopropylethylamine and 30mL of n-butanol to obtain a mixture; cooling the mixture to-20 ℃ by using a cold bath, slowly dropwise adding a compound a2(13.8mmol), reacting at a low temperature for 1h after the addition is finished, removing the cold bath, heating to room temperature, stirring overnight, decompressing and evaporating the solvent, adding 100mL of Ethyl Acetate (EA) into the residue, adding 50mL of water for washing for 2 times, evaporating an organic phase, and performing column chromatography separation on the residue (eluent is ethyl acetate: petroleum ether is 1:30 (volume ratio)), so as to obtain an intermediate A1.
The above process A1 relates to the following reaction formula:
wherein Ar is selected from phenyl or substituted phenyl, the substituted phenyl is mono-substituted, di-substituted or tri-substituted phenyl, and the substituents are respectively and independently selected from halogen, cyano, nitro, ester group and C1-4Alkyl or cycloalkyl, C1-4Alkoxy or cycloalkoxy, C1-4Haloalkyl, C1-4Acyl radical, C1-6Alkylamino or cycloalkylamino.
Method A2: adding 4.35mmol of the compound a1-2 into a 50mL single-neck bottle, adding 1mL of diisopropylethylamine and 20mL of n-butanol, adding 2.2mmol of the compound a2 to obtain a mixture, heating the mixture to 100 ℃, reacting overnight, evaporating the solvent under reduced pressure, adding 50mL of Ethyl Acetate (EA) into the residue, adding 50mL of water, washing for 2 times, evaporating the organic phase, and carrying out column chromatography separation on the residue to obtain an intermediate A2.
The above process A2 relates to the following reaction formula:
method A3: adding 9mmol of compound a1-3 into a 50mL single-neck bottle, adding 1mL of diisopropylethylamine and 25mL of n-butanol, adding 9mmol of compound a2 to obtain a mixture, heating the mixture to 100 ℃ for reaction overnight, evaporating the solvent under reduced pressure, adding 50mL of Ethyl Acetate (EA) into the residue, adding 50mL of water, washing for 2 times, evaporating the organic phase, and carrying out column chromatography separation on the residue (eluent is ethyl acetate: petroleum ether is 1:20 (volume ratio)), thus obtaining an intermediate A3.
The above process A3 relates to the following reaction formula:
the preparation of intermediate B is method B1 or method B2, as detailed below.
Method B1 includes the following steps:
1) synthesis of Compound b 1-2: 50g of compound b1-1 were dissolved in 250mL of tetrahydrofuran, 12.6g of sodium hydroxide were dissolved in 250mL of water, the two solutions were mixed and stirred overnight, the tetrahydrofuran was removed by rotary evaporation, the aqueous phase was washed twice with dichloromethane, most of the water was removed by rotary evaporation, the remainder was evaporated naturally, and the residue was dried thoroughly in a vacuum oven to give 55g of an orange solid, compound b 1-2. The analytical data for this compound are as follows: 1H-NMR (400MHz, & DMSO): δ: 7.73(t, J ═ 8.22, 1H); 6.02(dd, J ═ 2.97, 13.79, 1H); 5.77(dt, J ═ 2.87, 7.45, 1H).
2) Synthesis of Compound b 1-3: adding 7g of compound b1-2 and 17.5g of potassium carbonate into a reaction bottle, then adding 700mL of DMF (dimethylformamide), 70g of deuterium water and 17.5g of bromodifluoroacetic acid ethyl ester into the reaction bottle under the protection of argon, gradually heating to 50 ℃, stirring overnight, TLC shows that most of raw materials disappear, cooling, diluting the reaction liquid with water, extracting with dichloromethane for three times, combining organic phases, washing with water for 5 times, drying and spin-drying the organic layer, and separating the residue by column chromatography to obtain 6.2g of light yellow oily substance which is compound b 1-3. The analytical data for this compound are as follows: 1H-NMR (400MHz, CDCl)3)：δ：8.01(q,J＝5.58，3.51,1H)；7.07-7.15(m,2H)。
3) Synthesis of Compound b 1-4: dissolving 1g of compound b1-3 in 25mL of ethanol, adding palladium carbon, and stirring at room temperature and normal pressure in a hydrogen atmosphere overnight; TLC detection of disappearance of the starting material, filtration of the palladium on carbon, washing with ethanol and spin-drying of the solvent gave 0.78g of compound b1-4 as yellowA colored oil. The analytical data for this compound are as follows: 1H-NMR (400MHz, CDCl)3)：：δ：6.70-6.84(m,3H)；3.71(br,2H)；MS m/z(ESI):179[M+H]+。
4) Synthesis of Compound b 1-5: adding 0.78g of the compound b1-4 into 5mL of concentrated sulfuric acid cooled in an ice-water bath in batches at the temperature lower than 10 ℃ to completely dissolve the compound b1-4, then adding 0.45g of potassium nitrate, stirring at room temperature overnight, after the reaction is finished, pouring the reaction solution into ice water, then alkalifying with ammonia water, extracting an organic layer with ethyl acetate, combining the organic layers, washing with saturated saline solution once, drying and spin-drying the organic layer, and performing column chromatography separation on the residue to obtain 0.8g of yellow solid, namely the compound b 1-5. The analytical data for this compound are as follows: 1H-NMR (400MHz, CDCl)3)：δ：7.46(d,J＝6.48,1H)；6.99(d,J＝10.94,1H)；4.03(br,1H)；MS m/z(ESI):224[M+H]+。
5) Synthesis of Compound b 1-6: 3.1g of compound b1-5 and 171mg of DMAP (4-dimethylaminopyridine) are dissolved in 40mL of acetonitrile, 3.6g of (Boc)2O (di-tert-butyl dicarbonate) is added under ice bath, then the mixture is gradually raised to room temperature and stirred overnight, TLC detection raw materials disappear, then the reaction solution is directly dried by spinning, and the residue is subjected to column chromatography separation to obtain 3g of yellow solid, namely the compound b 1-6. The analytical data for this compound are as follows: 1H-NMR (400MHz, CDCl)3)：δ：8.99(d,J＝8.10,1H)；7.07(d,J＝12.15,1H)；6.85(br,1H)；MS m/z(ESI):324[M+H]+。
6) Synthesis of Compound b 1-7: dissolving 1.5g of compound b1-6, 950mg of N, N, N' -trimethylethylenediamine and 1.8g of diisopropyldiamine in 20mL of DMAC (N, N-dimethylacetamide), heating to 60 ℃, stirring overnight, detecting the disappearance of raw materials by TLC, pouring the reaction solution into water and Ethyl Acetate (EA), washing the water layer once with Ethyl Acetate (EA), then combining the EA layers, washing with water for 5 times to remove DMA, and drying the organic layer; the residue was subjected to column chromatography to give 2.2g of a yellow solid, i.e., compound b 1-7.
7) Synthesis of Compound b 1-8: 2.2g of compound b1-7 was dissolved in 40mL of Ethyl Acetate (EA), 0.2g of palladium on carbon was added thereto, the mixture was stirred overnight at room temperature under a hydrogen atmosphere, and the crude was detected by TLCWhen the material disappeared, the palladium-carbon was filtered off, and the mother liquor was spin-dried to obtain 2g of a yellow oily substance, i.e., compound b 1-8. The analytical data for this compound are as follows: 1H-NMR (400MHz, CDCl)3)：δ：7.47(s,1H)；6.78(s,1H)；6.69(br,1H)；4.25(br,2H)；2.85(t,J＝6.89,1H)；2.62(s,3H)；2.36(t,J＝6.48,1H)；2.25(s,6H)；MS m/z(ESI):376[M+H]+。
8) Synthesis of intermediate B-1: dissolving 2g of compound B1-8 in 40mL of dichloromethane, adding 0.83g of diisopropylethylamine, cooling to about 0 ℃ in an ice salt bath under the protection of nitrogen, then dropwise adding 0.55g of acryloyl chloride, gradually heating to room temperature after the dropwise addition, stirring for two hours, detecting the disappearance of raw materials by TLC, spin-drying the reaction solution, adding 5mL of concentrated hydrochloric acid into the residue, stirring for two hours, showing the disappearance of the raw materials by TLC, adjusting the pH to about 8 with a saturated sodium carbonate aqueous solution, alkalifying the system, extracting the reaction solution for three times with (ethyl acetate) EA, combining organic layers, drying, spin-drying, and carrying out column chromatography on the residue to obtain 750mg of yellow oily matter, namely an intermediate B1. The analytical data for this compound are as follows: 1H-NMR (400MHz, CDCl)3)：δ：8.07(s,1H)；6.92(s,1H)；6.39(dd,J＝1.63，17.09,1H)；6.22(dd,J＝10.48，17.47,1H)；5.68(dd,J＝1.42，9.91,1H)；3.81(br,2H)；2.77(t,J＝5.44,1H)；2.63(s,3H)；2.23(s,8H)；MS m/z(ESI):330[M+H]+。
Method B1 relates to the following reaction:
method B2 includes the following steps:
1) synthesis of Compound b 2-2: taking 50g of the compound b2-1, adding 500mL of methanol for complete dissolution, adding 10g of Pd/C (palladium on carbon), and carrying out hydrogenation reaction at 35 ℃ for two days; monitoring by a point plate, directly filtering Pd/C after the reaction of the raw materials is finished, and performing spin-drying on a methanol phase to obtain 39g of a crude compound b 2-2.
2) Synthesis of Compound b 2-3: 39g of crude compound b2-2 was added to 500mL of concentrated sulfuric acid (ice salt bath) at T<Stirring at 10 deg.C to dissolve completely, and adding while maintaining the temperature1ep of potassium nitrate, and stirring at room temperature overnight; the next day, the reaction mixture was poured into ice water, and the pH was adjusted with ammonia water>And 7, extracting with ethyl acetate, drying, and performing column chromatography separation to obtain 44g of a product, namely the compound b 2-3. The analytical data for this compound are as follows: 1H NMR (CDCl)3)δ7.39(d,J＝7.2Hz,1H),6.63(d,J＝12.4Hz,1H),3.94(s,3H),3.90(broad,2H)。
3) Synthesis of Compound b 2-4: adding 20g of compound b2-3 into 500mL of dichloromethane, cooling the mixture to-5 ℃ with an ice salt bath, dropwise adding 1.1eq of dichloromethane solution of di-tert-butyl dicarbonate, adding 0.2eq of DMAP (4-dimethylaminopyridine) after dropwise adding, naturally heating to room temperature, and stirring overnight; the next day, the reaction is performed by spotting, and after the reaction is completed, column chromatography separation is performed to obtain 24g of a yellow solid, namely the compound b 2-4. The analytical data for this compound are as follows: 1H NMR (CDCl)3)δ8.89(s,1H),6.97(s,1H),6.71(d,J＝12.4Hz,1H),3.97(s,3H),1.53(s,9H)；MS:Calcd for C12H15FN2O5(M-H)-:286.1,Found:285.0。
4) Synthesis of Compound b 2-5: taking 13.5g of the compound b2-4, adding the compound b2-4 into 200mL of DMAC (N, N-dimethylacetamide), and stirring to completely dissolve the compound b 2-4; then 2eq of N, N, N' -trimethylethylenediamine and 3eq of DIEA (N, N-diisopropylethylamine) were added, the temperature was raised to 110 ℃ and the mixture was stirred overnight; the next day, the reaction was complete to give 22g of crude compound b2-5 as an oil. The analytical data for this compound are as follows:1H NMR(CDCl3)δ8.54(s,1H),6.85(s,1H),6.60(s,1H),3.90(s,3H),3.22(t,J＝6.8Hz,2H),2.81(s,3H),2.55(t,J＝7.2Hz,2H),2.26(s,6H),1.49(s,9H)；MS:Calcdfor C17H28N4O5(M+H)+:368.21,Found:369.3。
5) synthesis of Compound b 2-6: 22g of crude compound b2-5 is taken and added into 400mL of ethyl acetate, stirred and dissolved completely, 4.07g of Pd/C (palladium on carbon) is added, and hydrogenation reaction is carried out at 20 ℃ overnight; the next day, after the reaction of the starting materials was complete, Pd/C was directly filtered off and concentrated to give 17g of crude compound b2-6 as a black oil. The analytical data for this compound are as follows:1H NMR(CDCl3)δ7.517(s,1H),6.941(s,1H),6.61(s,1H),4.10(m,2H),3.76(s,3H),2.92(m,2H),2.62(s,3H),2.40(m,2H),2.27(s,6H),1.49(s,9H)；MS:Calcd for C17H30N4O3(M+H)+:338.23,Found:339.4。
6) synthesis of Compound b 2-7: taking 17.3g of a crude compound b2-6, adding 500mL of dichloromethane and 1.2eq of DIEA (N, N-diisopropylethylamine), cooling to-5 ℃ in a ice salt bath, dropwise adding 1.1eq of acryloyl chloride under the protection of argon, naturally heating to room temperature after dropwise adding, after 3 hours, finishing the reaction, directly performing rotary evaporation and concentration at low temperature to remove the solvent, and obtaining 23g of a crude compound b 2-7.
7) Synthesis of intermediate B-2: 23g of crude compound b2-7 was taken and added to 50mL of THF, the ice salt bath was cooled to-5 deg.C, and 100mL of concentrated hydrochloric acid was added at temperature T<And (3) stirring the mixture for 2 hours at 10 ℃, performing plate spotting reaction, and performing column chromatography separation to obtain 5.2g of a product, namely an intermediate B2. The analytical data for this compound are as follows:1H NMR(CDCl3)δ10.10(s,1H),7.97(s,1H),6.68(s,1H),6.41-6.21(m,2H),5.65(m,1H),3.81(s,3H),3.76(s,2H),2.82(m,2H),2.65(s,3H),2.20(s,6H)；MS:Calcd for C15H24N4O2(M+H)+:292.19,Found:293.3。
method B2 relates to the following reaction:
method B3 includes the following steps:
1) synthesis of Compound b 3-2: adding 50g of the raw material compound b3-1 into 500mL of DMF, adding 1.5eq of ethyl difluorobromoacetate and 2eq of potassium carbonate, stirring at room temperature for 10min, adding 3eq of water, heating in an oil bath under the protection of argon to 50 ℃, cooling to 0 ℃ after 4 hours of reaction of the raw materials, adding water for quenching, extracting with an organic phase formed by mixing dichloromethane and petroleum ether in equal proportion, drying the organic phase, concentrating, and performing column chromatography separation to obtain 77g of a compound b3-2 product. The analytical data for this compound are as follows:1H NMR(CDCl3)δ8.03(dd,J＝9.2,5.6Hz,1H),7.149-7.081(m,2H),6.645(t,J＝72.4Hz,1H)。
2) transformingSynthesis of Compound b 3-3: taking 77g of compound b3-2, adding 700mL of absolute ethyl alcohol for complete dissolution, adding 13g of Pd/C (palladium on carbon), and carrying out hydrogenation reaction at 20 ℃ overnight; monitoring by a point plate, directly filtering Pd/C after the reaction of the raw materials is finished, and performing spin drying to obtain 55g of a crude compound b 3-3. The analytical data for this compound are as follows: 1H NMR (CDCl3) δ 6.84-6.70(m,3H),6.468(t, J ═ 73.6Hz,1H),3.16(broad, 2H); MS Calcd for C7H6F3NO(M+H)+:178.04,Found:178.00。
3) Synthesis of Compound b 3-4: 55g of the crude compound b3-3 was added to 600mL of concentrated sulfuric acid (ice salt bath), and<stirring at 10 ℃ for complete dissolution, keeping the temperature, adding 1ep potassium nitrate, and stirring at room temperature overnight; the next day, pour into ice water, adjust pH with ammonia>And 7, extracting with ethyl acetate, drying, and performing column chromatography separation to obtain 63g of a tawny product, namely the compound b 3-4. The analytical data for this compound are as follows:1H NMR(CDCl3)δ7.46(d,J＝7.2Hz,1H),7.01(d,J＝11.2Hz,1H),6.597(t,J＝72.4Hz,1H),4.047(broad,2H)。
4) synthesis of Compound b 3-5: adding 11.1g of compound b3-4 into 200mL of dichloromethane, cooling the mixture to-5 ℃ by using an ice salt bath, dropwise adding 1.1eq of dichloromethane solution of di-tert-butyl dicarbonate, adding 0.2eq of DMAP (4-dimethylaminopyridine) after dropwise adding, naturally heating to room temperature, stirring overnight, adding the mixture for the next day, spotting, carrying out reaction, and carrying out column chromatography separation to obtain 9.7g of a yellow product, namely the compound b 3-5. The analytical data for this compound are as follows: 1H NMR (CDCl)3)δ9.00(d,J＝8Hz,1H),7.07(d,J＝10.8Hz,1H),6.864(s,1H),6.661(t,J＝71.2Hz,1H),1.541(s,9H)；MS:Calcd for C12H13F3N2O5(M-H)-:323.08,Found:321.1。
5) Synthesis of Compound b 3-6: 0.82g of Compound b3-5 was dissolved in 10mL of DMAC (N, N-dimethylacetamide), and 2eq of R was added3H and 3eq of DIEA (N, N-diisopropylethylamine) are reacted at 80 ℃ overnight under the protection of argon; detecting the next day until the raw materials disappear; removing DMA by rotary evaporation at 80 deg.C, adding 50mL saturated sodium carbonate aqueous solution and 50mL dichloromethane, extracting with dichloromethane for 2 times, mixing organic phases, drying, and concentrating to obtain compound1.3g of crude product b 3-6. The analytical data for this compound are as follows: MS Calcd for C16H21F2N3O6(M+H)+:389.14,Found:390.2。
6) Synthesis of Compound b 3-7: dissolving 1.2g of the compound b3-6 in 100mL of ethyl acetate, adding 0.4eq of Pd/C (palladium on carbon), and carrying out hydrogenation reaction at 20 ℃ overnight; the detection of the next day shows that the raw material disappears, Pd/C is filtered out, the concentration is carried out, and the column chromatography separation is carried out, so that 0.7g of the compound b3-7 product is obtained. The analytical data for this compound are as follows: 1H NMR (CDCl)3)δ9.32(br,1H),9.23(s,1H),6.96(s,1H),6.73(s,1H),6.47(t,J＝74.8Hz,1H),5.85(m,1H),5.24(m,1H),3.86(m,4H),2.83(m,4H),1.53(s,9H)；MS:Calcd for C16H23F2N3O4(M+H)+:359.17,Found:360.2。
7) Synthesis of Compound b 3-8: taking 0.1g of raw material acrylic acid (or substituted acrylic acid), dissolving with 20mL of DCM (dichloromethane), adding 0.7eq of oxalyl chloride and one drop of DMF, and reacting at-5 ℃ for 4 hours under the protection of argon to generate acyl chloride to obtain an acyl chloride solution for later use; meanwhile, in another reaction flask, 0.2g (0.5eq) of compound b3-7 was taken out, dissolved in 20mL of dry dichloromethane, and 0.15g of DIEA was added thereto, and the prepared acid chloride solution was poured into the reaction mixture at-5 ℃ for reaction overnight; the next day, directly spin-drying to obtain compound b 3-8. The analytical data for this compound are as follows: MS Calcd for C19H24F3N3O5(M+H)+:431.17,Found:432.2。
8) Synthesis of intermediate B-3: taking 0.1g of compound b3-8, adding 3mL of concentrated hydrochloric acid, discharging bubbles when adding, and stirring for 3 minutes; the reaction solution was added dropwise to 10mL of saturated sodium carbonate solution, and after dropping, the solution pH>10; extracting with 30mL of dichloromethane for 2 times, drying and concentrating to obtain 0.08g of a product, namely an intermediate B3. The analytical data for this compound are as follows: MS Calcd for C14H16F3N6O6(M+H)+:331.11,Found:332.1。
Method B3 relates to the following reaction:
example 1
The structural formula of the pyrimidine compound of the embodiment is shown as formula I-1:
the preparation method of the pyrimidine compound of the embodiment is as follows: dissolving 50mg (0.15mmol) of the intermediate B, 150mg (0.5mmol) of the intermediate A and 35mg (0.18mmol) of p-toluenesulfonic acid monohydrate in 5mL of 2-pentanol, heating to 50 ℃, stirring overnight under the protection of nitrogen, TLC (thin layer chromatography) for basically disappearance of raw materials and volume spinning, adding 20mL of dichloromethane and 20mL of saturated aqueous sodium carbonate solution, layering, washing the aqueous phase twice with 20mL of dichloromethane, combining the organic phases, drying and spinning, and separating by TLC to obtain 20mg of a product, namely the compound I-1. The analytical data for this compound are as follows: 1H-NMR (400MHz, CDCl)3)：δ：10.18(br,1H)；9.19(br,1H)；8.38(s,1H)；7.52(dt,J＝2.39，11.32,1H),7.16-7.24(m,2H)；7.03-7.06(m,2H)；6.86(br,1H)；6.71(t,J＝7.11,1H)；6.30-6.38(m,2H)；5.67(dd,J＝2.62，9.58,1H)；2.84(t,J＝5.27,2H)；2.70(s,3H)；2.37(t,J＝4.91,2H)；2.30(s,6H)；MS m/z(ESI):585[M+H]+。
The preparation method relates to the following reaction formula:
wherein, the intermediate A1-1 is prepared by the method A1; intermediate B1-1 was prepared using method B1 described above.
The pyrimidine compounds of examples 2 to 41 and the intermediates A and B used in the preparation method are shown in Table 1, and the rest is the same as example 1.
TABLE 1 pyrimidines of examples 2-41 and intermediates A, B used in their preparation
Example 42
The pyrimidine compound of this example is the mesylate of the pyrimidine compound (I-6) shown in example 6, and the structural formula is shown in formula I-42:
the preparation method of the pyrimidine compound (mesylate) of the embodiment comprises the following steps: adding 0.37g of the compound I-6 into a 50mL single-neck bottle, adding 10mL of acetone and 1mL of water, after the addition, slowly adding 64mg of methanesulfonic acid under stirring, after the addition, reacting at 50 ℃ for 3h, after the reaction liquid is evaporated to dryness, adding 6mL of acetonitrile, heating to 70 ℃, stirring for 30min, slowly cooling to separate out a solid, filtering the solid, washing with acetonitrile, and drying to obtain 140mg of a white solid, namely the compound I-30. The purity was determined by HPLC to be 98.5%. The analytical data for the compound I-30 obtained were: HNMR (400M, d)6-DMSO):9.35(s1H),9.14(s,1H),8.76(s,1H),8.70(s,1H),8.36(s,1H),7.95(s,2H),7.85(d,J＝8.10Hz,1H),7.41(m,2H),6.97(s,1H),6.60(m,1H),6.24(d,J＝16.7Hz,1H)，5.78(d,J＝11.4Hz,1H),3.82(s,3H),3.26(s,4H),2.79(s,6H),2.59(s,3H),2.31(s,3H)。
The preparation method relates to the following reaction formula:
examples of the experiments
First, this experimental example examined the inhibitory activity of the pyrimidine compounds of examples 1 to 41 on wild-type EGFR and mutant EGFR kinase.
The method is used for determining the inhibition effect of the substance to be detected on the activity of double mutant EGFR kinase (EGFR T790M/L858R kinase) and wild type EGFR kinase (EGFR WT). Wild type EGFR and mutant EGFR (T790M/L858R) kinases used in this assay were purchased from Carna Bioscience.
Experiment design:
preparation of test compounds:
1. the compound to be tested was formulated as a 10mM (mmol/L) DMSO solution and the control compound AZD9291 was formulated as a 1mM (mmol/L) DMSO solution.
2. Test compound solutions were serially diluted to 12 concentrations (or other desired test concentrations) by 3-fold dilution on 384-well plates of a TECAN EVO 200.
3. Transfer 20nL of test solution to 384-well plates using Echo550 (Coring 3570). DMSO was used as a blank control.
Carrying out an enzyme test:
1. a1.3 Xenzyme solution containing the enzyme, substrate and cofactor was prepared as shown in Table 2 below.
2. mu.L of 1.3 Xenzyme solution was added to each well of the well plate and incubated at room temperature for 30 minutes.
3. The test reaction was started by adding 5. mu.L of 4 XATP solution. The final volume of solution in each well should be 20. mu.L, containing the ingredients shown in Table 2 below.
4. The plates were incubated at room temperature for 90 minutes and the test reactions were stopped by adding 40. mu.L of stop buffer (containing 0.5M EDTA).
5. Experimental data for each well was analyzed using EZ detection.
TABLE 2 enzyme solution parameter Table in enzyme test
And (3) data analysis:
1. using the read Conversion (CR), the inhibition ratio was calculated according to the following formula:
2. IC50 and Ki values were calculated using XLFit (equalisation 201) according to the following formula,
the results of the measurements are shown in Table 3 below.
TABLE 3 results of Activity inhibition assays for wild-type EGFR and mutant EGFR kinases
By combining the experimental results, compared with the prior art, the pyrimidine compound provided by the invention has the following advantages:
(1) the compounds shown in formula I have very good inhibitory activity on EGFR, particularly the inhibitory activity on EGFR mutation (particularly EGFR T790M/L858R mutation) which is obviously higher than that of AZD9291 in the prior art, such as examples 1, 3, 9, 11, 12, 14, 15, 16, 17, 18, 19, 20, 21, 22 and the like. On the premise of achieving the same treatment effect, the dosage can be greatly reduced, so that other side effects caused by the medicine are greatly reduced.
(2) The compound shown in the formula I has low inhibitory activity on wild EGFR, is obviously superior to a compound AZD9291 in the prior art of the same generation, such as examples 1, 2, 3, 4, 6, 8 and the like, has selective inhibitory activity on enzyme up to 10-60 times, is obviously superior to 3 times of the compound AZD9291 in the prior art, and has higher selectivity than a second generation EGFR inhibitor. In the aspect of medicine application, the problems of serious toxic and side effects and the like of rash of patients caused by strong inhibition on wild EGFR can be well reduced.
(3) The compounds of the invention also exhibit advantageous physical properties (e.g., water solubility, etc.), advantageous metabolic profiles (e.g., better pharmacokinetic profiles, such as bioavailability), compared to other known EGFR mutation inhibitors.
Secondly, the compound is used for testing the cell growth inhibition activity:
the test methods and procedures are performed by methods well known to those skilled in the art and the reagents used in the methods are commercially available.
The test method comprises the following steps:
1. the experimental steps are as follows:
(1) 40nL of test compound solution was applied to the test plate using Echo (non-contact nanoliter sonic pipetting system).
(2) Cells were prepared as a 25000cell/mL solution, and 40 μ L was then plated onto the designated 384-well test plate.
(3) The plates were incubated at 37 ℃ in 5% carbon dioxide at 95% humidity for 72 hours.
(4) Adding 40 μ L of the extract to each well
And (3) a reagent.
(5) The test plate was incubated at room temperature for 30 minutes to stabilize the luminescence signal.
(6) The test plate was sealed and air bubbles were removed at a centrifugation speed of 1000 rpm.
(7) The test plate was shaken on a shaker for 1 minute.
(8) The test board data is read.
2. Data processing
(1) The residual ratio was calculated using the following formula:
s: testing the sample reading;
v: reading a blank sample;
m: AZD9291 assay (1. mu.M for PC-9 and H1975, 30. mu.M for A431) readings;
IC50 was calculated using XLFIT (V5.3.1.3) software.
The results are shown in Table 4.
TABLE 4 results of the measurement of cytostatic Activity of Compounds
As can be seen from Table 4, the exemplary compounds of the present invention showed strong inhibitory activity against EGFR mutant cells (H1975, PC-9); compared with a control compound AZD9291, the inhibitory activity of the compound on the growth of EGFR mutant cells is improved by 4-7 times compared with the control compound AZD 9291.
Wherein, the structure of a comparison compound AZD9291 (trade name: Merritinib) is as follows:
Claims (7)
1. a pyrimidine compound, characterized in that: a compound comprising the structural formula:
2. a pyrimidine compound according to claim 1, wherein: the pharmaceutically acceptable salt is an inorganic acid salt or an organic acid salt, and the inorganic acid salt is selected from hydrochloride, hydrobromide, hydroiodide, sulfate, bisulfate, nitrate, phosphate and acid phosphate; the organic acid salt is selected from formate, acetate, trifluoroacetate, propionate, pyruvate, glycolate, oxalate, malonate, fumarate, maleate, lactate, malate, citrate, tartrate, methanesulfonate, ethanesulfonate, benzenesulfonate, salicylate, picrate, glutamate, salicylate, ascorbate, camphorate, camphorsulfonate.
3. An EGFR inhibitor characterized by: comprising a pyrimidine compound according to any one of claims 1 to 2.
4. The EGFR inhibitor according to claim 3, wherein: also comprises a pharmaceutically acceptable carrier.
5. Use of an EGFR inhibitor according to claim 3 for the preparation of a medicament for modulating EGFR tyrosine kinase activity or treating EGFR related diseases.
6. Use according to claim 5, characterized in that: the EGFR tyrosine kinase activity regulation or EGFR related disease treatment refers to cancer, diabetes, immune system diseases, neurodegenerative diseases or cardiovascular diseases.
7. Use of an EGFR inhibitor according to claim 3 for the preparation of a medicament for the treatment of non-small cell lung cancer.
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| CA3145686C (en) * | 2019-07-26 | 2023-03-21 | Charles Z. Ding | Pyrimidine compound acting on egfr and erbb2 |
| WO2021104305A1 (en) * | 2019-11-26 | 2021-06-03 | 上海翰森生物医药科技有限公司 | Nitrogen-containing polycyclic derivative inhibitor, preparation method therefor and application thereof |
| CN114539269B (en) * | 2020-11-19 | 2023-04-28 | 上海医药工业研究院有限公司 | Nitrogen-containing macrocyclic compound, preparation method and application thereof |
| CN117177968A (en) * | 2021-03-11 | 2023-12-05 | 昂科比克斯有限公司 | Pyrimidine derivatives exhibiting inhibitory effect on cancer cell growth |
| CN117794902A (en) * | 2021-08-02 | 2024-03-29 | 迪哲(江苏)医药股份有限公司 | Novel pharmaceutically acceptable salts and polymorphic forms of ErbB and BTK inhibitors |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103501612A (en) * | 2011-05-04 | 2014-01-08 | 阿里亚德医药股份有限公司 | Compounds for inhibiting cell proliferation in EGFR-driven cancers |
| CN105377835A (en) * | 2013-07-11 | 2016-03-02 | 贝达药业股份有限公司 | Protein tyrosine kinase modulators and methods of use |
| CN105384694A (en) * | 2014-08-22 | 2016-03-09 | 四川海思科制药有限公司 | Substituted aminopyrimidine derivative, preparation method therefor and pharmaceutical application thereof |
| CN105601573A (en) * | 2014-11-24 | 2016-05-25 | 中国科学院上海药物研究所 | 2-aminopyrimidine compounds as well as pharmaceutical compositions and applications thereof |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP6469567B2 (en) * | 2012-05-05 | 2019-02-13 | アリアド・ファーマシューティカルズ・インコーポレイテッド | Compound for inhibiting cell proliferation of EGFR-activated cancer |
| CN105503827B (en) * | 2014-10-11 | 2019-09-24 | 上海翰森生物医药科技有限公司 | EGFR inhibitor and its preparation method and application |
| CN106661000B (en) * | 2014-10-11 | 2019-04-09 | 上海翰森生物医药科技有限公司 | EGFR inhibitor and its preparation and application |
-
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103501612A (en) * | 2011-05-04 | 2014-01-08 | 阿里亚德医药股份有限公司 | Compounds for inhibiting cell proliferation in EGFR-driven cancers |
| CN105377835A (en) * | 2013-07-11 | 2016-03-02 | 贝达药业股份有限公司 | Protein tyrosine kinase modulators and methods of use |
| CN105384694A (en) * | 2014-08-22 | 2016-03-09 | 四川海思科制药有限公司 | Substituted aminopyrimidine derivative, preparation method therefor and pharmaceutical application thereof |
| CN105601573A (en) * | 2014-11-24 | 2016-05-25 | 中国科学院上海药物研究所 | 2-aminopyrimidine compounds as well as pharmaceutical compositions and applications thereof |
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| CN106083736A (en) | 2016-11-09 |
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