CN105153122A

CN105153122A - [(indole-3-yl)pyrimidine-2-yl]aminophenylpropyl-2-eneamide derivative and its salt, preparation method of derivative, and application of derivative and salt

Info

Publication number: CN105153122A
Application number: CN201510534307.4A
Authority: CN
Inventors: 彭快; 郑飞鸣; 傅勇
Original assignee: Shanghai Sheng Kao Pharmaceutical Technology Co Ltd
Current assignee: Henan Inno Medicine Technology Co ltd
Priority date: 2015-08-27
Filing date: 2015-08-27
Publication date: 2015-12-16
Anticipated expiration: 2035-08-27
Also published as: CN105153122B

Abstract

The invention provides a [(indole-3-yl)pyrimidine-2-yl]aminophenylpropyl-2-eneamide derivative and its salt, a preparation method of the derivative, and application of the derivative and the salt. The [(indole-3-yl)pyrimidine-2-yl]aminophenylpropyl-2-eneamide derivative has a structure shown in the formula I below. Deuterium-carbon bonds in the derivative enable the derivative to decompose slowly in a human body, a medicament of the derivative has a longer half-life period and a higher concentration in blood, the dosage of the medicament is finally reduced, and toxic and side effects of the medicament are decreased. Experiments show that compared with AZD 9291 mesylates, AZD 929-D9 mesylate of the deuterium-substituted derivative has Cmax which is 1.32 times as high as that of AZD 9291, exposed dose 1.41 times as high as that of AZD 9291, and elimination half-life 1.31 times as long as that of AZD 9291.

Description

[ (indol-3-yl) pyrimidine-2-yl ] aminophenylpropyl-2-enamide derivative and salt, preparation method and application

Technical Field

The invention relates to the technical field of medicinal chemistry, in particular to [ (indole-3-yl) pyrimidine-2-yl ] aminophenylpropyl-2-enamide derivatives and salts thereof, and a preparation method and application thereof.

Background

Worldwide, the first malignancy to be both morbidity and mortality is lung cancer, the most common of which is non-small cell lung cancer. Epidermal growth factor receptor-1 (EGFR) is usually expressed on the surface of non-small cell lung cancer tumor cells; upon binding to its ligand or forming heterodimers with other epidermal growth factor receptors (HER2, EGFR3), downstream signaling pathways in the cell may be activated, leading to abnormal proliferation of the cell. It can be seen that EGFR is a very important therapeutic target for non-small cell lung cancer. In particular, in some non-small cell lung cancers, such as non-squamous cell carcinomas, there is a 19 exon point mutation and a 21 exon L858R deletion mutation of EGFR. Such EGFR-resistant Tyrosine Kinase Inhibitors (TKIs), such as Gefitinib (Gefitinib), Erlotinib (Erlotinib), Afatinib (Afatinib), are particularly effective in such patients. Compared with chemotherapy, the three medicines have high effective rate, no disease and long progressive period, and low toxicity.

However, tumors bearing EGFR sensitive mutations develop secondary mutations in EGFR at position 20, T790M, resulting in secondary resistance to such TKIs, which occurs at about 50% of the time during treatment with such TKIs. In addition, some patients carry the primary EGFR RT790M mutation, which is resistant to this type of TKI. Therefore, in order to overcome the drug resistance to Gefitinib (Gefitinib), Erlotinib (Erlotinib) and Afatinib (Afatinib) caused by secondary and primary drug resistance of the EGFRT790M, an irreversible EGFR tyrosine kinase inhibitor is developed, has good inhibition effect on non-small cell lung cancer with T790M mutation, 19 exon point mutation and L858R point mutation of EGFR, and can become a second-line therapeutic drug after Gefitinib (Gefitinib), Erlotinib (Erlotinib) and Afatinib (Afatinib) drug resistance.

In the prior art, AZD-9291 is a highly efficient selective EGFR mutant inhibitor; the molecular formula is C₂₈H₃₃N₇O₂(ii) a The structural formula is shown as formula 1:

the IC50 of AZD9291 was 12.92nM, 11.44nM and 493.8nM for exon 19 deleted EGFR, L858R/T790MEGFR and wild type EGFR, respectively. However, the half-life of AZD9291 is short, and the dosage is high, which causes the occurrence rate of adverse metabolism, such as high diarrhea, rash and nausea, therefore, the structure of the drug needs to be optimized to increase the half-life, so as to reduce the dosage of the drug and reduce the toxicity of the drug.

Disclosure of Invention

In view of the above, the present invention aims to provide [ (indol-3-yl) pyrimidin-2-yl ] aminophenylprop-2-enamide derivatives and salts thereof, and a preparation method and an application thereof, and the [ (indol-3-yl) pyrimidin-2-yl ] aminophenylprop-2-enamide derivatives provided by the present invention have an increased half-life, reduce the administration dosage of a drug, and reduce the drug toxicity.

The present invention provides [ (indol-3-yl) pyrimidin-2-yl ] aminophenylprop-2-enamide derivatives having the structure of formula I:

in the formula I, R is₁、R₂、R₃、R₄、R₅、R₆、R₇、R₈、R₉、R₁₀、R₁₁、R₁₂、R₁₃、R₁₄、R₁₅、R₁₆、R₁₇、R₁₈、R₁₉And R₂₀Independently hydrogen or deuterium.

Preferably, the number of deuterium atoms in the formula I is 9-29.

Preferably, the derivative has the structure of formula 101, formula 102, formula 103, or formula 104:

the invention provides a mesylate of the derivative in the technical scheme, which has a structure shown in a formula II:

the invention provides a preparation method of the derivative in the technical scheme, which comprises the following steps:

reacting a compound with a structure shown in a formula III with a compound with a structure shown in a formula IV to obtain a compound with a structure shown in a formula V;

sequentially reducing and amidating the compound with the structure of the formula V to obtain a derivative with the structure of the formula I;

the R is₄、R₅、R₆、R₇、R₈、R₉、R₁₀、R₁₁、R₁₂、R₁₃、R₁₄、R₁₅、R₁₆、R₁₇、R₁₈、R₁₉And R₂₀Independently selected from hydrogen or deuterium;

in the formula III, M is F, Cl, Br or I.

Preferably, the compound having the structure of formula III is prepared by the following preparation method:

reacting a compound with a structure shown in a formula VI with a compound with a structure shown in a formula VII to obtain a compound with a structure shown in a formula III;

the R is₁₁、R₁₂、R₁₃、R₁₄、R₁₅、R₁₆、R₁₇、R₁₈、R₁₉And R₂₀Independently selected from hydrogen or deuterium;

in the formula VI, R is F, Cl, Br or I.

Preferably, the temperature for the reaction of the compound with the structure of formula VI and the compound with the structure of formula VII is 90-100 ℃; the reaction time is 18-22 h.

Preferably, the reaction temperature of the compound with the structure of formula III and the compound with the structure of formula IV is 93-98 ℃; the reaction time is 5.5-6.5 h.

The invention provides an application of the derivative in the technical scheme, the mesylate of the derivative in the technical scheme or the derivative prepared by the preparation method in the technical scheme in the preparation of a medicament for treating tumors.

Preferably, the tumor is a malignant tumor.

The present invention provides [ (indol-3-yl) pyrimidin-2-yl)]An aminophenylpropyl-2-enamide derivative having the structure of formula I. The invention provides [ (indol-3-yl) pyrimidin-2-yl)]Deuterium-carbon bonds in the aminophenyl propyl-2-enamide derivatives lead the decomposition speed in vivo to be slow, prolong the half-life period of the medicine, increase the blood concentration of the medicine, finally reduce the dosage and reduce the toxic and side effect. The experimental results show that: compared with AZD9291 mesylate, C, the deuterated derivative AZD9291-D9 mesylate provided by the invention_maxIs 1.32 times of AZD9291 mesylate; its exposure (AUC)_0-t) Is 1.41 times of AZD9291 mesylate; it has elimination half-life t_1/2Is 1.31 times of AZD9291 mesylate, namely is prolonged by 31 percent,

drawings

FIG. 1 is a reaction scheme of example 1 of the present invention;

FIG. 2 is a LC-MS detection scheme of a brown solid obtained by preparation of example 1, step nine, of the present invention;

FIG. 3 is a NMR chart of a brown solid obtained by the ninth preparation in example 1 of the present invention;

FIG. 4 is a NMR chart of a brown solid obtained by the ninth preparation in example 1 of the present invention;

FIG. 5 is an LC-MS detection scheme of an off-white solid obtained by step ten preparation of example 1 of the present invention;

FIG. 6 is a NMR spectrum of a white-like solid obtained by the tenth preparation in example 1 of the present invention;

FIG. 7 is a graph showing the results of testing nude mice with the mesylate drug prepared in the examples and comparative examples of the present invention;

FIG. 8 shows the results of the test of the mesylate drug prepared in the examples and comparative examples for 40-120 days in nude mice.

Detailed Description

In the invention, the number of deuterium atoms in the formula I is preferably 9-29; in a specific embodiment of the present invention, the number of deuterium atoms in said formula I is specifically 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28 or 29.

In the present invention, the derivative preferably has the structure of formula II, formula III, formula IV or formula V:

in the invention, the number of deuterium atoms in the formula II is preferably 9-29; in a particular embodiment of the invention, the number of deuterium atoms in said formula II is in particular 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28 or 29.

In the present invention, the mesylate salt of the derivative preferably has the structure of formula 201, formula 202, formula 203, or formula 204:

in the formula III, M is F, Cl, Br or I.

The invention reacts the compound with the structure of formula VI and the compound with the structure of formula VII to obtain the compound with the structure of formula V.

In the invention, the compound with the structure of the formula III is prepared by the following preparation method:

in the formula VI, R is F, Cl, Br or I.

The source of the compound having the structure of formula VI is not particularly limited in the present invention, and the compound having the structure of formula VI known to those skilled in the art can be used, for example, commercially available products thereof can be used, and the compound having the structure of formula VI can be prepared by itself by using the technical scheme known to those skilled in the art for preparing the compound having the structure of formula VI. In the present invention, the compound having the structure of formula VI is preferably prepared by the following method:

reacting the compound with the structure of the formula VII and the trideuteroiodomethane to obtain a compound with the structure of the formula VI;

the source of the compound having the structure of formula VII in the present invention is not particularly limited, and the compound having the structure of formula VII known to those skilled in the art can be used, for example, commercially available compounds thereof can be used, or the compound having the structure of formula VII can be prepared by themselves using the technical scheme known to those skilled in the art for preparing the compound. In the present invention, the compound having the structure of formula VII is preferably prepared by the following preparation method:

reacting a compound with a structure of formula VIII with a compound with a structure of formula IX to obtain a compound with a structure of formula VII;

in the present invention, the compound having the structure of formula VIII is preferably specifically represented by formula 301:

in the present invention, the compound having the structure of formula IX is preferably specifically represented by formula 302:

in the invention, the compound with the structure of the formula VIII is preferably dissolved in dichloromethane to obtain a compound solution with the structure of the formula VIII. The volume ratio of the substance of the compound with the structure of the formula VIII to dichloromethane is preferably 0.13-0.16 mol: 500 mL. Preferred in the invention is AlCl₃In the presence of a compound of the structure of formula VIII and a compound of the structure of formula IX; the AlCl₃And a compound of formula VIIIThe mass ratio of the substances is preferably 0.15-0.2: 0.13-0.15.

In the present invention, the mass ratio of the compound of the structure of formula VIII to the compound of the structure of formula IX is preferably 1: 1; the reaction temperature of the compound with the structure of formula VIII and the compound with the structure of formula IX is preferably 20-30 ℃; the reaction time of the compound with the structure of formula VIII and the compound with the structure of formula IX is preferably 12-15 h. In the present invention, the reaction solution is preferably subjected to extraction. The invention preferably adopts ethyl acetate EtOAc for extraction; the number of extractions is preferably 3.

The reaction of the compound having the structure of formula VII and trideuteroiodomethane is preferably carried out in a nitrogen atmosphere. The present invention preferably performs the reaction of the compound having the structure of formula VII and trideuteroiodomethane in the presence of NaH and tetrahydrofuran. In the invention, the mass ratio of the NaH, the compound with the structure of the formula VII and the trideuteroiodomethane is preferably (6.7-6.8): (5.5-5.6): (6.6-6.7), more preferably (6.73-6.78): (5.53-5.55) and (6.63-6.68). In the present invention, the temperature at which the compound having the structure of formula VII and trideuteroiodomethane react is preferably 0 ℃; the time for the reaction of the compound having the structure of formula VII and trideuteroiodomethane is preferably 1.5 hours. In the invention, the reaction liquid obtained by the reaction of the compound with the structure of the formula VII and the trideuteroiodomethane is preferably extracted. The invention preferably adopts ethyl acetate EtOAc for extraction; the number of extractions is preferably 3. In the present invention, the organic phase obtained by extraction is preferably washed. The invention preferably employs a saturated brine wash; the number of washing is preferably 3. The invention preferably dries the washed product; the invention preferably adopts anhydrous sulfate for drying; the anhydrous sulfate is preferably anhydrous sodium sulfate.

In the present invention, the compound having the structure of formula VI is preferably specifically represented by formula 303:

the present invention is not particularly limited with respect to the source of the compound having the structure of formula VII, and any compound having the structure of formula VII known to those skilled in the art may be used, for example, commercially available compounds thereof may be used. In the present invention, the compound having the structure of formula VII is preferably specifically represented by formula 304:

the reaction of the compound having the structure of formula VI and the compound having the structure of formula VII is preferably carried out in the presence of p-toluenesulfonic acid and sec-amyl alcohol according to the present invention. In the present invention, the mass ratio of the compound having the structure of formula VI, the compound having the structure of formula VII, and p-toluenesulfonic acid is preferably 1: 1.15 to 1.2; the volume of the secondary amyl alcohol and the quantity ratio of the substances of the p-toluenesulfonic acid are preferably (23-26) mL: (1.15-1.2) mol. In the invention, the temperature for the reaction of the compound with the structure of formula VI and the compound with the structure of formula VII is preferably 90-100 ℃, and more preferably 95 ℃; the reaction time of the compound with the structure of formula VI and the compound with the structure of formula VII is preferably 18-22 h, and more preferably 20 h. In the invention, the reaction liquid obtained by the reaction of the compound with the structure of the formula VI and the compound with the structure of the formula VII is preferably cooled to room temperature, and solid is precipitated. The invention preferably carries out suction filtration, washing and drying on the obtained solid in sequence. The washing is preferably carried out using sec-amyl alcohol and water in this order. Vacuum drying is preferably employed in the present invention.

In the present invention, the compound having the structure of formula III is preferably specifically represented by formula 305:

the source of the compound having the structure of formula IV is not particularly limited in the present invention, and the compound having the structure of formula IV can be prepared by those skilled in the art, for example, commercially available products thereof can be used, or the compound having the structure of formula IV can be prepared by themselves by those skilled in the art. In the present invention, the preparation method of the compound having the structure of formula IV preferably comprises the following steps:

reacting the compound with the structure of the formula X with deuterated dimethylamine to obtain a compound with the structure of the formula XI;

reducing the compound with the structure of the formula XI to obtain a compound with the structure of the formula IV;

the compound with the structure of the formula X is reacted with deuterated dimethylamine to obtain the compound with the structure of the formula XI. In the present invention, the compound having the structure of formula X is preferably specifically represented by formula 306:

in the present invention, the deuterated dimethylamine preferably participates in the reaction in the form of a hydrochloride of deuterated dimethylamine; the mass ratio of the compound with the structure of the formula X to the deuterated dimethylamine is preferably 34-35: 22 to 23. In the present invention, the temperature of the reaction of the compound having the structure of formula X and deuterated dimethylamine is preferably 20 ℃ to 30 ℃; the reaction time of the compound with the structure of the formula X and the deuterated dimethylamine is preferably 20-24 h.

After the compound with the structure of the formula XI is obtained, the compound with the structure of the formula XI is reduced by the invention to obtain the compound with the structure of the formula IV.

The present invention preferably employs lithium aluminum hydride for the reduction of the compound having the structure of formula IV. In the invention, the mass ratio of the lithium aluminum hydride to the compound with the structure of formula IV is preferably 9-11: 1, and more preferably 10: 1. The present invention preferably performs the above reduction in the presence of anhydrous tetrahydrofuran. In the invention, the temperature of the reduction is preferably 0-5 ℃; the reduction time is preferably 5.5-6.5 h. The present invention preferably quenches, filters, adjusts the pH value, extracts and dries the reaction solution. The present invention preferably employs water for quenching. In the present invention, the pH of the reaction solution is preferably adjusted to 2 to 3. In the invention, the reaction solution after pH value adjustment is preferably mixed with water and then extracted to obtain a water phase. In the invention, the aqueous phase is preferably dried by freeze drying.

In the present invention, the compound having the structure of formula IV is preferably specifically represented by formula 307:

in the present invention, the reaction of the compound having the structure of formula III and the compound having the structure of formula IV is preferably performed in the presence of N, N-Diisopropylethylamine (DIEA) and N, N-Dimethylacetamide (DMAC). In the invention, the reaction temperature of the compound with the structure of formula III and the compound with the structure of formula IV is preferably 93-98 ℃; the reaction time of the compound with the structure of the formula III and the compound with the structure of the formula IV is preferably 5.5-6.5 h. In the invention, the mass ratio of the compound with the structure of formula III to the compound with the structure of formula IV is preferably 7-9: 10.

According to the invention, the reaction liquid obtained by the reaction of the compound with the structure of the formula III and the compound with the structure of the formula IV is preferably cooled to 20-30 ℃. In the present invention, the reaction solution after temperature reduction is preferably mixed with water, and stirred to precipitate a solid. According to the invention, the reaction liquid with solid separated out is preferably subjected to suction filtration to obtain a filter cake; washing the filter cake; sequentially extracting, washing, drying and concentrating the filtrate to obtain a solid product; and mixing the solid product and the filter cake to obtain the compound with the structure of the formula V.

After the compound with the structure of the formula V is obtained, the compound with the structure of the formula V is sequentially reduced and amidated to obtain the derivative with the structure of the formula I. The reduction of the compound of formula V is preferably carried out in the presence of iron powder and ammonium chloride. The invention preferably carries out the reduction of the compound with the structure of the formula V in a mixed solution of ethanol and water. According to the invention, the compound with the structure of formula V is heated to reflux for reaction; the reduction time of the compound with the structure shown in the formula V is preferably 2.5-3.5 h, and more preferably 3 h. According to the invention, the reaction liquid obtained by reducing the compound with the structure of the formula V is preferably cooled and filtered; and concentrating the obtained filtrate, and purifying the concentrate to obtain a reduction product.

The invention amidates the reduction product to obtain the derivative with the structure of formula I. The present invention preferably utilizes 3-chloropropionyl chloride and NaOH for amidation of the reduction product. Preferably, the reduction product is firstly reacted with 3-chloropropionyl chloride to obtain an intermediate; and reacting the intermediate with NaOH to obtain the derivative with the structure of the formula I. In the present invention, the amidation is preferably performed in a mixed solvent of tetrahydrofuran and water. In the invention, the mass ratio of the reduction product to the 3-chloropropionyl chloride is preferably 0.8-0.9: 1. In the present invention, the temperature of the amidation is preferably 0 ℃; the amidation time is preferably 9-11 h. Preferably, the amidation product is cooled to room temperature and then is mixed with water to obtain a mixed solution; extracting the mixed solution to obtain an organic phase; and washing, drying, decompressing, concentrating and purifying the organic phase in sequence. The present invention preferably employs ethyl acetate for extraction. The present invention preferably employs anhydrous sulfate for drying. The present invention preferably uses methanol and dichloromethane in a volume ratio of 1:10 as eluent for purification.

In the present invention, the mesylate of the derivative has the structure of formula II. The process for preparing the methanesulfonate salt of the derivative of the present invention is not particularly limited, and a method for forming the salt of the methanesulfonate salt, which is well known to those skilled in the art, may be used. In the present invention, the method for preparing the methanesulfonic acid salt of the derivative preferably comprises the steps of:

reacting the derivative with methanesulfonic acid to obtain the methanesulfonic acid salt of the derivative.

In the present invention, the derivative is preferably dissolved in a mixed solution of acetone and water to obtain a derivative solution. In the present invention, the ratio of the amount of the derivative to the volume of the mixed solution is preferably (0.07 to 0.09) mol: (0.4 to 0.5) mL, more preferably 0.08 mol: 0.44 mL; the volume ratio of acetone to water is preferably 10: 1. In the present invention, the methanesulfonic acid preferably participates in the reaction as an acetone solution of methanesulfonic acid; the volume ratio of the quantity of the methanesulfonic acid and the substance to the acetone is preferably (0.07-0.09) mmol: (0.045 to 0.055), more preferably 0.08 mmol: 0.05 mL.

In the invention, the reaction temperature of the derivative and methanesulfonic acid is 55-65 ℃; the reaction time of the derivative and methanesulfonic acid is preferably 1-5 hours.

In the present invention, it is preferable that the reaction solution obtained by reacting the derivative with methanesulfonic acid is cooled to 20 to 30 ℃ and a white solid is precipitated. In the present invention, it is preferred that the cooled reaction solution is filtered, and the obtained solid is washed with methyl tert-butyl ether (MTBE) to obtain the methanesulfonate of the derivative.

In the present invention, pharmaceutically acceptable salts, crystalline hydrates, solvates, prodrugs, single crystals or polymorphs of the derivatives are all within the scope of the present invention.

The invention also provides application of the derivative in the technical scheme, mesylate of the derivative in the technical scheme or the derivative prepared by the preparation method in the technical scheme in preparing a medicament for treating tumors.

In the present invention, the tumor is preferably a malignant tumor; the malignant tumor preferably comprises medullary thyroid carcinoma, non-small cell lung cancer, pancreatic cancer, renal cancer, metastatic renal cell carcinoma, bladder cancer, ovarian cancer, brain cancer, breast cancer, prostate cancer, multiple myeloma, soft tissue sarcoma, melanoma cancer, recurrent/metastatic Merkel cell carcinoma, endometrial cancer, plexiform neurofibroma, liver cancer, advanced cholangiocellular carcinoma, adult glioblastoma, metastatic nodal carcinoma, or acute myelogenous leukemia. More preferably medullary thyroid cancer, prostate cancer, liver cancer, or kidney cancer. In some embodiments of the invention, the malignancy targeted is non-small cell lung cancer.

In the invention, when the derivative or the methanesulfonate of the derivative is used as a medicine for treating tumors, the derivative or the methanesulfonate of the derivative further comprises pharmaceutically acceptable auxiliary materials. In the present invention, the pharmaceutically acceptable excipients include: one or more of pharmaceutically acceptable carriers, excipients, diluents, adjuvants, vehicles.

Pharmaceutically acceptable carrier materials include, but are not limited to: sugars such as lactose, glucose and sucrose; starches such as corn starch and potato starch; serum proteins, such as human serum albumin; cellulose and its derivatives such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; buffer substances, such as phosphates, glycine, sorbic acid, potassium sorbate; ion exchangers, aluminum stearate lecithin; partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as protamine sulfate, sodium chloride, disodium hydrogenphosphate, polyvinylpyrrolidone, colloidal silica, potassium hydrogenphosphate, magnesium trisilicate, polyacrylates, zinc salts, lanolin, polyethylene-polyoxypropylene-blocking polymers; gum powder; malt; gelatin; talc powder; adjuvants such as cocoa butter and suppository waxes; oils such as peanut oil, safflower oil, cottonseed oil, sesame oil, olive oil, soybean oil and corn oil; glycols, such as propylene glycol and polyethylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffering agents such as aluminum hydroxide and magnesium hydroxide; alginic acid; pyrogen-free water; isotonic salt; ringer's solution; ethanol, phosphate buffered solutions, and other non-toxic suitable lubricants such as sodium lauryl sulfate and magnesium stearate, coloring agents, releasing agents, coating materials, sweetening, flavoring and perfuming agents, preservatives and antioxidants.

In the present invention, the tumor drug prepared from the derivative or the mesylate salt of the derivative may be administered orally, by injection, by inhalation spray, topically, rectally, nasally, buccally, vaginally, or via an implantable kit. In the present invention, the injection administration includes subcutaneous, intravenous, intramuscular, intraarticular, intrasynovial (luminal), intrasternal, intramembranous, intraocular, intrahepatic, intralesional, intracranial injection or infusion techniques. In the present invention, the mode of administration of the drug is preferably oral administration, intraperitoneal administration, or intravenous injection. In the present invention, the pharmaceutical compositions may be administered in the form of aqueous or oleaginous suspensions which may be formulated according to the known art using suitable dispersing, wetting and suspending agents. The sterile injectable preparation may be a sterile injectable solution or suspension in a non-toxic acceptable diluent or solvent, for example, a solution of 1, 3-butanediol. These acceptable excipients and solvents may be water, sodium chloride solution, ringer's solution, and the like. Further, sterile, nonvolatile oils may conventionally be employed as a solvent or suspending medium.

Solid dosage forms for oral administration may include tablets, capsules, powders, granules and pills. Further, in these dosage forms, the active compound is mixed with at least one pharmaceutically acceptable inert excipient or carrier or filler, such as sodium citrate, calcium phosphate. Fillers such as lactose, starch, glucose, mannitol, sucrose or silicic acid; binders such as alginates, carboxymethylcellulose, sucrose, polyvinylpyrrolidone, gelatin, or acacia; absorbents such as bentonite or kaolin; humectants such as glycerol; disintegrating agents such as calcium carbonate, potato starch, alginic acid, agar-agar, tapioca starch, certain silicates or sodium carbonate; absorption enhancers such as quaternary ammonium compounds; wetting agents such as cetyl alcohol and glyceryl monostearate; lubricants such as solid polyethylene glycol, magnesium stearate, calcium stearate, talc, sodium lauryl sulfate, and mixtures thereof; retarder solutions such as paraffin; for dosage forms such as tablets, capsules or pills, buffering agents may be included.

Solid dosage forms may include dragees, tablets, granules, pills, capsules, and the like, and may be prepared using coatings or shells, such as enteric coatings or other materials well known in the art. And the active compound in such compositions can be released in a delayed manner, in a certain part of the digestive tract. Further, the active compound may be in microencapsulated form with one or more of the excipients mentioned above.

The tumor drug may also be incorporated into a suitable delivery vehicle that provides controlled and/or continuous release of the compound, and may also serve as a targeting moiety. Examples of delivery vehicles include, without limitation, adjuvants, synthetic adjuvants, microcapsules, microparticles, liposomes, and yeast cell wall particles. The walls of the yeast cells can be variously treated to selectively remove protein components, glucan or mannan layers.

The oncology drug may be administered orally in any acceptable oral dosage form including, but not limited to: capsules, tablets, aqueous suspensions or aqueous solutions. For oral administration as tablets and enteric coated tablet dosage forms, fillers generally include lactose, sucrose, corn starch, lactose, glucose, silicic acid, sodium citrate, mannitol, dicalcium phosphate; binders such as starch, cellulose derivatives, gelatin, polyvinylpyrrolidone (pvp), sucrose; lubricants such as glycerin; disintegrating agents such as agar, calcium carbonate, potato, tapioca starch, alginic acid, or croscarmellose sodium; lubricants such as magnesium stearate and glycerin are typically added. For oral administration in capsules, suitable diluents include lactose and dried corn starch. When the oral administration is an aqueous suspension, the active ingredient thereof consists of an emulsifying agent and a suspending agent. In preparing these dosage forms, pharmaceutically acceptable sweetening, flavoring or coloring agents may also be added.

In the present invention, the dose of the tumor drug prepared from the derivative or the mesylate of the derivative should be determined in consideration of the administration route, the health condition of the patient, and the like. In the present invention, the daily dose of the drug to be administered to a person having a body weight of 60kg is preferably 1 to 1000mg, more preferably 5 to 100 mg.

To further illustrate the present invention, the [ (indol-3-yl) pyrimidin-2-yl ] aminophenylprop-2-enamide derivatives and salts thereof, the preparation method and the use thereof provided by the present invention will be described in detail with reference to the following examples, which should not be construed as limiting the scope of the present invention.

Example 1

Referring to the reaction scheme shown in FIG. 1, FIG. 1 is a reaction scheme of example 1 of the present invention:

the method comprises the following steps: preparation of 3- (2-Chloropyrimidin-4-yl) -1H-indole

N₂To a solution of Compound 1(21g,0.14mol) in dichloromethane (500mL) under an atmosphere, AlCl was added in portions₃(22.5g,0.17mol), 10 minutes was added. After the resulting suspension was stirred at room temperature for 10 minutes, the reaction system was substantially clear. Compound 2(16.5g,0.14mol) was added to the above reaction mixture in portions at room temperature over 10 minutes. The resulting brown suspension was stirred at room temperature overnight; the reaction was monitored by TLC, and when complete consumption of compound 1 was achieved, the reaction was poured into ice water (500mL) with stirring and the system was extracted 3 times with EtOAc (300 mL). The organic phases were combined and washed 2 times with brine (200mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure to give 38g of a brown viscous crude product. The crude product was purified by silica gel column chromatography (eluent n-hexane: EtOAc ═ 5:1 to 3:1) to give 16g of the title compound 3 as a yellow solid in 49% yield.

The LC-MS test results for compound 3 were: LCMS (ESI +): M/z230.15(M + H);¹H-NMR(300MHz,DMSO-d6)12.09(brs,1H),8.59-8.54(m,2H),8.44-8.40(m,1H),7.92(d,J＝5.4Hz,1H),7.52-7.50(m,1H),7.34-7.20(m,2H)。

step two: preparation of 3- (2-chloropyrimidin-4-yl) -1-trideuteromethyl-indole

Under the nitrogen atmosphere and under the cooling of ice bath, 60% NaH (0.27g,6.75mmol) is added in portions to a THF (50mL) solution of 3- (2-chloropyrimidin-4-yl) -1H-indole 3(1.27g,5.53mmol), the reaction solution is stirred for 0.5 hour at 0 ℃, trideuteroiodomethane (0.96g,6.64mmol) is added dropwise to the reaction system, the reaction is finished after 10min, and the reaction is carried out for 1.5 hours at 0 ℃, and the reaction is monitored by TLC; when the starting material 3 was consumed, the reaction was terminated, and a saturated sodium bicarbonate solution (10mL) was slowly dropped into the reaction mixture to quench the reaction. The resulting mixture was extracted 3 times with EtOAc (40mL), the organic phases were combined and washed 3 times with saturated brine (40 mL); after drying over anhydrous sodium sulfate, concentration under reduced pressure gave 1.29g of yellow solid 4 in 94.9% yield, which was used in the next step without further purification of compound 4.

The test results for compound 4 were: 96.8 percent of HPLC; LCMS (ESI +): M/z247.16(M + H);¹H-NMR(300MHz,CDCl₃)8.46-8.42(m,1H),8.38-8.31(m,1H),7.95(s,1H),7.50-7.46(m,1H),7.40-7.28(m,3H)。

step three: preparation of N- (4-fluoro-2-methoxy-5-nitrophenyl) -4- (I-trideuteromethyl-indol-3-yl) pyrimidin-2-amine

Compound 4(1.19g,4.82mmol), compound 5(0.90g,4.82mmol) and p-TsOH were reacted at room temperature^.H₂O (1.1g,5.72mmol) was added to sec-amyl alcohol (24mL), and the resulting suspension was replaced with nitrogen gas 3 times, stirred for 5 minutes, and then warmed to 95 ℃ for reaction. The reaction was monitored by HPLC. After 20 hours HPLC showed complete disappearance of starting compound 4 and the reaction was complete. After the reaction mixture was cooled to room temperature, it was stirred for 1 hour to precipitate a solid. Suction was applied and the filter cake was washed successively with sec-amyl alcohol (2mL) and water (2 mL). The resulting solid was dried under vacuum to give 1.8g of compound 6 as a yellow solid. The product was used in the next step without further purification.

The test results for compound 6 were: HPLC 94.8% (minus p-TsOH); LCMS (ESI +): M/z397.11(M + H);¹H-NMR (300MHz, DMSO-d6)9.84(brs,1H),8.75(d, J ═ 8.1Hz,1H),8.69(t, J ═ 2.1Hz,1H),8.32(d, J ═ 6.6Hz,1H),8.20(d, J ═ 7.2Hz,1H),7.59(t, J ═ 8.1Hz,1H),7.53-7.47(m,2H),7.31(t, J ═ 7.5Hz,1H),7.13(t, J ═ 8.1Hz,1H),4.00(s,3H) (crude, containing approximately 25% TsOH).

Step four: preparation of intermediate 7-2

To the reaction flask were added 7(10g,0.13mol,1.0eq), 50mL of water and 50mL (1mol/L) of aqueous sodium hydroxide solution. Cooling to 0-5 ℃ by using an ice water bath after stirring and clarification, then dropwise adding 7-1(14.4g,0.13mol,1.0eq) toluene solution (30mL), and supplementing sodium hydroxide aqueous solution (1M, 216mL) during dropwise adding to keep the pH value of the system between 8-10. After the addition was complete, the mixture was allowed to warm to room temperature and stirred for 30 minutes. The reaction was diluted with 500mL of water and extracted with EtOAc (250mLx 3). The aqueous phase was collected and adjusted to pH 3-4 with dilute hydrochloric acid (1mol/L) and extracted three times with extraction followed by EtOAc (250mLx 3). The EtOAc phase is Na₂SO₄Drying, filtering and concentrating to obtain the compound 7-2.

¹HNMR(300MHz,CDCl₃)5.28(s,2H),4.20-4.15(m,2H),4.13-4.02(m,2H),1.27(t,3H,J＝7.2Hz)。

Step five: preparation of intermediates 7-3

To a solution of Pvi-Cl (4.13g,34.27mmol,1.5eq) and TEA (4.0g,39.53mmol,1.7eq) in chloroform (55mL) at room temperature was slowly added dropwise a solution of compound 7-2(5.04g,34.27mmol,1.5eq) in chloroform (55 mL). After the dropwise addition, the mixture was stirred at room temperature for 1.5 hours under nitrogen protection. Under nitrogen protection, hydrochloride of deuterated dimethylamine (2.0g, 22.84mmol,1.0eq) was added, followed by stirring at room temperature for 22 hours in a closed system. 100mL of water was added, and the mixture was stirred at room temperature for 1 hour, 40mL of chloroform was added, and the organic phase was washed once with 100mL of water, 3 times with 100mL of an aqueous solution of sodium carbonate (1mol/L), and once with 100mL of water. The chloroform phase is treated with Na₂SO₄Drying, filtering, concentrating and purifying by column chromatography to obtain compound 7-3(3.0g, yield: 72%);

¹HNMR(300MHz,CDCl₃)5.68(s,1H),4.21-4.10(m,2H),4.02-3.98(m,2H),1.27(t,3H,J＝7.2Hz).

step six: preparation of N-methyl-N ', N' -bis (trideuteromethyl) 1, 2-ethanediamine 8

Compound 7-3(3.0g,16.6mmol,1.0eq) was dissolved in 100mL of anhydrous THF. And (3) cooling to 0-5 ℃ by using an ice water bath under the protection of nitrogen, then slowly adding lithium aluminum hydride (7.0g,166mmol,10eq) in batches, and slowly heating to micro reflux after no bubbles are generated stably. The reaction was complete after 6 hours. And cooling the system to room temperature, cooling to 0-5 ℃ by using an ice water bath, dropwise and slowly adding 7mL of water for quenching, dropwise and slowly adding 7mL of 20% sodium hydroxide aqueous solution, and dropwise and slowly adding 7mL of water. After stirring for 30 min at rt, filtration and washing the filter cake three times with THF, the filtrate was collected, the solution was evaporated to dryness after adjusting the pH2-3 with dilute hydrochloric acid, water (150mL) was added and extracted three times with EtOAc (150mLx 3). The aqueous phase was lyophilized by freezing to give Compound 8(2.6g, yield: 86%).

The test results for compound 8 were:¹HNMR(300MHz,D₂O)3.51-3.42(m,4H),2.74(s,3H)。

step seven: preparation of N '- (2- [ bis (trideuteromethyl) amino ] ethyl) -2-methoxy-N' -methyl-N- [4- (1-trideuteromethyl-indol-3-yl) pyrimidin-2-yl ] -5-nitrobenz-1, 4-diamine 9

Adding compound 6(300mg,0.76mmol), compound 8(170.0mg,0.94mmol) and DIEA (440.0mg,3.4mmol) into DMAC (3mL) at room temperature, heating to 95 ℃ in a sealed tank for reaction, monitoring the reaction by HPLC, and after 6 hours, completely disappearing the raw material 6 by HPLC to finish the reaction; cooling the reaction solution to room temperature, adding water (3mL), stirring at room temperature for 2 hours, separating out solids, performing suction filtration, and washing a filter cake with water (2 mL); the filtrate was extracted 3 times with EtOAc (30mL), and the organic phases were combined and washed 6 times with water (30mL) to remove the DMAC. Dried by anhydrous sodium sulfate, and concentrated under reduced pressure to obtain brick red solid compound 9 with yield of 73%. The product was used in the next step without further purification.

The test results for compound 9 were: LCMS (ESI +): M/z485.33(M + H);¹H-NMR (crude 300MHz, CDCl)₃)9.59(s,1H),8.41-8.18(m,4H),7.63(s,1H),7.41-7.20(m,2H),7.20(d,J＝5.1Hz,1H),6.82(s,1H),4.04(s,3H),3.30(t,J＝6.9Hz,2H),3.0(s,3H),2.60(t,J＝6.9Hz,2H)。

Step eight: preparation of N¹- (2- [ di (trideuteromethyl) amino)]Ethyl) -5-methoxy-N¹-methyl-N⁴- [4- (1-Trideuteromethyl-indol-3-yl) pyrimidin-2-yl]Benzene-1, 2, 4-triamine 10

Compound 9(1.08g,2.23mmol), iron powder (0.750g,13.38mmol) and NH were added at room temperature₄Cl (0.084g,1.57mmol) was added to EtOH/H₂O (60mL/20mL) and warmed to reflux. After 3 hours, TLC showed complete disappearance of starting material 9 and the reaction was complete; cooling the reaction solution to room temperature, and filtering; the filter cake was washed 3 times with ethanol (10mL), the filtrate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography (eluent MeOH: DCM ═ 1:10, 1% aqueous ammonia) to give 790mg of compound 10 as a brown-gray solid in 78% yield.

The test results for compound 10 were: 98.1 percent of HPLC; LCMS (ESI +): M/z455.33(M + H);¹H-NMR(300MHz,CDCl₃)8.48-8.46(m,1H),8.34(d,J＝5.1Hz,1H),8.20(s,1H),7.82(s,1H),7.64(s,1H),7.39-7.28(m,3H),7.05(d,J＝5.1Hz,1H),6.67(s,1H),3.86(s,3H),3.49(s,2H),3.30(t,J＝5.7Hz,2H),3.08(d,J＝5.7Hz,2H),2.72(s,3H)

step nine: preparation of N- (2- {2- [ bis (trideuteromethyl) amino ] ethyl- (methyl) amino } -4-methoxy-5- { [4- (1-trideuteromethylindol-3-yl) pyrimidin-2-yl ] amino } phenyl) prop-2-enamide (AZD9291-d9)

To compound 10(0.7g,1.54mmol) in THF/H with ice-cooling₂3-chloropropionyl chloride (0.24g,1.85mmoL) is dripped into an O (6mL/0.6mL) solution, and dripping is completed after 10 minutes; the reaction solution was further stirred at 0 ℃. After 1 hour, TLC and LCMS showed complete consumption of starting material 10; NaOH (0.25g,6.15mmoL) was added to the reaction solution, and the temperature was raised to reflux. After 9 h, LCMS and TLC showed the intermediate was essentially reacted, the reaction was allowed to cool to rt, water (1mL) was added and extracted 3 times with EtOAc (30mL), the organic phases were combined and washed 3 times with water (10 mL). Dried over anhydrous sodium sulfate and concentrated under reduced pressure to give 0.9g of crude brown viscous substance. The crude product was purified by silica gel column chromatography (eluent MeOH: DCM ═ 1:10) to give 0.61g of foamy brown solid compound 11 in 78% yield.

LC-MS detection is carried out on the compound obtained in the ninth step, the detection result is shown in figure 2, figure 2 is an LC-MS detection diagram of the brown solid prepared in the ninth step of the example 1, and the LC-MS detection diagram can be seen from figure 2: LCMS (ESI +): M/z509.30(M + H);

in the invention, the compound obtained in the ninth step is subjected to nuclear magnetic resonance hydrogen spectrum detection, the detection result is shown in fig. 3, fig. 3 is a nuclear magnetic resonance hydrogen spectrum of the brown solid prepared in the ninth step in example 1 of the invention, and as can be seen from fig. 3:¹H-NMR(300MHz,CDCl₃)10.21(brs,1H),9.87(s,1H),9.14(s,1H),8.40(d,J＝5.4Hz,1H),8.09-8.07(m,1H),7.74(s,1H),7.43-7.40(m,1H),7.32-7.23(m,2H),7.22(d,J＝5.4Hz,1H),6.82(s,1H),6.52-6.35(m,2H),5.72(dd,J₁＝2.4Hz,J₂＝9.6Hz,1H),3.96(s,3H),2.91(t,J＝5.4Hz,2H),2.73(s,3H),2.30-2.28(m,2H)。

according to the invention, the nuclear magnetic resonance carbon spectrum test is performed on the compound obtained in the ninth step, the test result is shown in fig. 4, and fig. 4 is a nuclear magnetic resonance carbon spectrum of the brown solid prepared in the ninth step in example 1 of the invention.

Step ten: preparation of N- (2- {2- [ bis (trideuteromethyl) amino ] ethyl- (methyl) amino } -4-methoxy-5- { [4- (1-trideuteromethylindol-3-yl) pyrimidin-2-yl ] amino } phenyl) prop-2-enamide methanesulfonate (AZD9291-d9 methanesulfonate)

Dissolving compound 11(40mg,0.08mmoL) in acetone/water (0.4mL/0.04mL), heating to 60 deg.C, adding a solution of methanesulfonic acid (7.6mg,0.08mmoL) in acetone (0.05mL) at one time, stirring the reaction solution at 60 deg.C for 1.5 hr, naturally cooling to room temperature to precipitate off-white solid, filtering, washing with MTBE (1mL), and drying to obtain 44.6mg of white solid 12(AZD9291-d9 methanesulfonate); the yield of the off-white solid AZD9291-d9 mesylate was 94%.

Chemical purity > 98%, deuteration purity > 99%;

LC-MS detection is carried out on the off-white solid obtained in the tenth step of the example 1, the detection chart is shown in FIG. 5, FIG. 5 is the LC-MS detection chart of the off-white solid AZD9291-d9 mesylate prepared in the tenth step of the example 1, and as can be seen from FIG. 5: LCMS (ESI +): M/z509 (M-OMs);

the invention performs nuclear magnetic resonance hydrogen spectrum test on the off-white solid obtained in the step ten, the test result is shown in fig. 6, and fig. 6 is a nuclear magnetic resonance hydrogen spectrum of the off-white solid AZD9291-d9 mesylate prepared in the step ten of the example 1 of the invention.

As can be seen from fig. 6:¹H-NMR(300MHz,DMSO-d6)9.60(s,1H),9.18(s,1H),8.78(s,1H),8.54(s,1H),8.32(d,J＝5.7Hz,2H),7.96(s,1H),7.53(d,J＝8.1Hz,1H),7.27-7.13(m,3H),7.02(s,1H),6.69(dd,J₁＝9.9Hz,J₂＝16.5Hz,1H),6.33(d,J＝16.5Hz,1H),5.82(d,J＝10.2Hz,1H),3.91(s,3H),3.28(brs,4H),2.63(s,3H),2.09(s,3H)。

1.1 in vitro experiments on the deuterated derivative AZD9291-D9 mesylate

AZD9291-D9 mesylate prepared in example 1 was taken and examined for its killing effect on PC-9 and PC-9 resistant strain cells, with AZD9291 mesylate and gefitinib as positive controls and PBS as a blank control.

20000 cells of PC-9 and PC-9 resistant strains were inoculated to 7 75cm cells²And (4) carrying out cell culture in a cell culture flask. Subculturing the cells at 37 ℃, a 5% carbon dioxide incubator, counting, centrifuging at 1000rpm for 5 minutes, adjusting the cell density by using a culture medium, and inoculating the cells in a 96-well plate according to 2000 cells/100 mu L/well; on the next day, 100. mu.g/mL, 30. mu.g/mL, 10. mu.g/mL, 3. mu.g/mL, 1. mu.g/mL, 0.3. mu.g/mL, 0.1. mu.g/mL, 0.03. mu.g/mL, 0.01. mu.g/mL were each dosed, and PC-9 cells and PC-9 drug-resistant strains were cultured for 3 days and 5 days, respectively, according to the above cell doubling time. On the detection day, 100 μ L of CCK8 reagent (Nippon Hojindo chemical) is added into each cell culture well, the cell culture wells are placed in an incubator and kept stand for 1 hour, the light absorption value is measured at 450nm by a Beijing Pulang enzyme labeling instrument (model DNM9602), GraphPad software is applied to calculate IC50, and the detection result of the killing effect of the sample to be detected on the PC-9 and PC-9 drug-resistant strains is shown in Table 1; and gefitinib to PC-9The results of the detection of the killing effect of the drug-resistant strains are shown in table 2:

TABLE 1 test results of the killing effect of the samples to be tested on PC-9 and PC-9 resistant strains

TABLE 2 detection results of the killing effect of gefitinib on PC-9 resistant strains

As can be seen from Table 1, the samples to be tested in the experimental group and the positive control group have equivalent killing effect on the cells of the PC-9 drug-resistant strain and half-growth inhibitory concentration IC₅₀The deuterated AZD9291-D9 mesylate prepared in example 1 of the invention has equivalent killing effect on cancer cells to AZD9291 mesylate (P > 0.05), namely the in vitro bioactivity of the AZD9291 mesylate is not changed after being deuterated.

1.2 pharmacokinetic evaluation of the deuterated derivative AZD9291-D9

The deuterated AZD9291-D9 mesylate prepared in example 1 was taken and subjected to pharmacokinetic experiments with AZD9291 mesylate as a positive control and PBS as a blank control.

The tested animals were: male CD1 mice weighing 18-22 g were provided by Shanghai drug laboratory animal center, and were licensed as SYXK (Shanghai) 2010-0049. The test animals are adaptively raised in a test place 3-7 days before the test day, male CD1 mice are randomly divided into 2 groups, the mice are respectively subjected to gastric lavage and are given with the tested samples, the mice are fasted for 12 hours before the test, the mice freely drink water, and the mice are uniformly fed after being given for 2 hours; the samples to be tested are given 25mg/kg dose by single gavage, and the samples to be tested of the experimental group and the positive control group use 0.5% w/v hydroxypropyl methylcellulose (HPMC);

dissolving 0.1% w/v Tween in water; sampling at 0.25h, 0.5h, 1.0h, 2.0h, 3.0h, 5.0h, 8.0h, 10h and 24h after administration; 3 mice at each time point, 0.3mL of venous blood is taken from retrobulbar venous plexus of the mice after animal anesthesia at the set time points, the venous blood is placed into a heparinized test tube, 11000g of the venous blood is centrifuged for 5 minutes, and plasma is separated and frozen in a refrigerator at the temperature of 20 ℃ below zero; during sample detection, after protein is precipitated by methanol, plasma samples are subjected to LC-MS/MS (liquid chromatography-mass spectrometry) to determine the concentrations of AZD9291 mesylate and deuterated AZD9291-D9 mesylate in plasma, wherein the linear range is 30.0-30000 ng/mL.

The WinNonlin6.3 software was used to calculate the major pharmacokinetic parameters (T) after gastric gavage in mice_max，C_maxAUC, MRT and t_1/2). Wherein the peak concentration C is reached_maxAnd time to peak T_maxIs an actual measurement value.

Area under plasma concentration-time curve AUC_0-tThe value: and calculating by adopting a trapezoidal method.

AUC_0-∞＝AUC_0-t+C_t/k_e，

C_tThe blood concentration at the last measurable time point, k_eTo eliminate the rate constant;

elimination of half-life t_1/2＝0.693/k_e；

The mean residence time MRT is AUMC/AUC.

After mice were gavaged with 25mg/kg of AZD9291-D9 mesylate and AZD9291 mesylate, respectively, the pharmacokinetic parameters of the drugs are shown in Table 3, and Table 3 shows the pharmacokinetic parameters of the mice after gavaged with 25mg/kg of drugs, respectively:

TABLE 3 pharmacokinetic parameters of mice after gavage with 25mg/kg drug, respectively

Slave watch3, the experimental data show that the preparation method provided in example 1 prepares the C of deuterated AZD9291-D9 mesylate_maxIs 1.32 times of AZD9291 mesylate; its exposure (AUC)_0-t) Is 1.41 times of AZD9291 mesylate; it has elimination half-life t_1/2The deuterated AZD9291-D9 mesylate is 1.31 times of AZD9291 mesylate, namely is prolonged by 31%, and the deuterated AZD9291-D9 mesylate prepared by the invention is remarkably improved in blood concentration and prolonged in half-life period compared with AZD9291 mesylate and has remarkable difference.

Example 2

Preparation of N- (2- {2- [ bis (trideuteromethyl) amino ] ethyl- (methyl) amino } -4-methoxy-5- { [4- (1-trideuteromethyl-2, 4,5,6, 7-pentadeuteroindol-3-yl) pyrimidin-2-yl ] amino } phenyl) prop-2-enamide methanesulfonate (AZD9291-D14 methanesulfonate)

Example 2 was prepared according to the method described in example 1, except that: 1H-indole-2, 3,4,5,6,7-d6 is used for replacing 1H-indole to prepare the target product.

Through the test: chemical purity > 98%, deuteration purity > 98%; LCMS (ESI +): M/z514 (M-OMs).

Comparative example 1

Preparation of N- (2- {2- [ bis (trideuteromethyl) amino ] ethyl- (methyl) amino } -4-methoxy-5- { [4- (1-methylindol-3-yl) pyrimidin-2-yl ] amino } phenyl) prop-2-enamide methanesulfonate (AZD9291-D6 methanesulfonate):

the preparation was as described in example 1, except that: and replacing the trideuteroiodomethane with iodomethane to obtain the target product.

Chemical purity > 98%, deuteration purity > 99%; LCMS (ESI +): M/z506 (M-OMs).

Comparative example 2

Preparation of N- (2- { 2-dimethylaminoethyl- (methyl) amino } -4-methoxy-5- { [4- (methyl-2, 4,5,6, 7-pentadeuterated indol-3-yl) pyrimidin-2-yl ] amino } phenyl) prop-2-enamide methanesulfonate (AZD9291-D5 methanesulfonate):

the preparation was as described in example 1, except that: the target product is prepared by replacing trideuteroiodomethane with iodomethane, replacing N-methyl-N ', N' -dimethyl-1, 2-ethylenediamine with N-methyl-N ', N' -di (trideuteromethyl) 1, 2-ethylenediamine with 1H-indole-2, 3,4,5,6,7-d6 with 1H-indole.

Chemical purity > 98%, deuteration purity > 99%; LCMS (ESI +): M/z508 (M-OMs).

Comparative example 3

Preparation of N- (2- { 2-dimethylaminoethyl- (methyl) amino } -4-methoxy-5- { [4- (1-trideuteromethylindol-3-yl) pyrimidin-2-yl ] amino } phenyl) prop-2-enamide methanesulfonate (AZD9291-D3 methanesulfonate):

the preparation was as described in example 1, except that: the N-methyl-N ', N' -di (trideuteromethyl) 1, 2-ethanediamine 8 is replaced by N-methyl-N ', N' -dimethyl-1, 2-ethanediamine, so as to prepare the target product.

Chemical purity > 98%, deuteration purity > 99%; LCMS (ESI +): M/z503 (M-OMs).

Example 3 nude mouse experiment of mesylate salt of deuterated derivative:

culturing PC-9 lung cancer cell in RPMI1640 culture medium, and collecting 5 × 10 cells⁶(volume: 0.1mL) cells were inoculated subcutaneously into BALB/C-NU nude mice and grown for 4 months; measuring the growth condition of the transplanted tumor every 2 weeks, and calculating the tumor volume, wherein when the volume of the transplanted tumor grows to 0.2-0.4 cm³At this time, mice were randomized according to tumor size and divided into treatment and placebo control groups, respectively. Mean changes in tumor volume were measured using intragastric administration/placebo daily and counted using a two-sided T-test.

A model of the transplanted tumors was constructed and gavage fed with 2mg/kg body weight of AZD9291, AZD9291-D3 prepared in comparative example 1, AZD9291-D5 prepared in comparative example 2, AZD9291-D6 prepared in comparative example 1, AZD9291-D9 prepared in example 1, AZD9291-D14 prepared in example 2 and PBS negative control, respectively, 6 tumors per group were measured at different time points, and the tumor volume was measured in cm³。

7 groups of animals, PC-9 cell nude mouse model are set: the drug concentrations of the blank group, AZD9291 mesylate, AZD9291-D3 mesylate, AZD9291-D5 mesylate, AZD9291-D6 mesylate, AZD9291-D9 mesylate and AZD9291-D14 mesylate are all 2 mg/kg.

The test results for nude mice are shown in fig. 7, and fig. 7 is a test result for nude mice for the mesylate drug prepared in the examples and comparative examples of the present invention; wherein,the test curves of the blank group to the nude mice are shown,is a test curve of AZD9291 mesylate to nude mice,is a test curve of AZD9291-D5 mesylate to nude mice,the test curves of AZD9291-D3 to nude mice are shown,is a test curve of AZD9291-D6 mesylate to nude mice,is a test curve of AZD9291-D9 mesylate to nude mice,the test curve of AZD9291-D14 mesylate to nude mice is shown. FIG. 8 is an enlarged view of a portion of a curve in FIG. 7, and FIG. 8 is a result of testing 40-120 days on nude mice with methanesulfonate prepared according to examples and comparative examples of the present invention, wherein,is a test curve of AZD9291 mesylate to nude mice,the test curves of AZD9291-D3 to nude mice are shown,is a test curve of AZD9291-D6 mesylate to nude mice,is a test curve of AZD9291-D9 mesylate to nude mice,is a test curve of AZD9291-D14 mesylate to nude mice,the test curve of AZD9291-D5 mesylate to nude mice is shown.

The experimental data in FIG. 7 and FIG. 8 show that the tumor-bearing nude mice have the inhibiting ability, and AZD9291-D14 is AZD9291-D9, AZD9291-D6, AZD9291-D3, AZD9291-D5 and AZD9291 blank group; AZD9291-D9 is 1.5 times of AZD9291-D6 and 1.8 times of AZD 9291-D3; is 2 times of AZD 9291. In contrast, AZD9291-D14 and AZD9291-D9 showed the strongest inhibitory effect on tumor-bearing nude mice tumors.

The mesylate of the deuterated derivative provided by the invention greatly increases the half-life of the drug and prolongs the time of the drug staying in a human body; meanwhile, the concentration of the medicine in the blood is improved, so that a better curative effect is achieved; compared with AZD9291 mesylate, the mesylate of the deuterated derivative provided by the invention has smaller dosage, so that the problem of poor metabolism of the drug can be further solved, and the drug toxicity and other side effects can be reduced.

As can be seen from the above examples, the present invention provides [ (indol-3-yl) pyrimidin-2-yl ] aminophenylprop-2-enamide derivatives, having the structure of formula I. The deuterium-carbon bond in the [ (indole-3-yl) pyrimidine-2-yl ] aminophenylpropane-2-enamide derivative provided by the invention ensures that the deuterium-carbon bond is low in decomposition speed in vivo, the half-life period of the medicament is prolonged, the blood concentration of the medicament is increased, the dosage is finally reduced, and the toxic and side effects are reduced.

In addition, the derivative provided by the invention is more stable in vivo, reduces the metabolism of the derivative in the inner wall of the intestinal tract and the liver, reduces the undesirable metabolism of the medicament, and further reduces the toxicity and other side effects of the medicament.

The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.

Claims

[ (indol-3-yl) pyrimidin-2-yl ] aminophenylprop-2-enamide derivatives having the formula I:

in the formula I, R is₁、R₂、R₃、R₄、R₅、R₆、R₇、R₈、R₉、R₁₀、R₁₁、R₁₂、R₁₃、R₁₄、R₁₅、R₁₆、R₁₇、R₁₈、R₁₉And R₂₀Independently hydrogen or deuterium.
2. The derivative according to claim 1, wherein the number of deuterium atoms in formula I is 9-29.
3. The derivative of claim 1, wherein the derivative has the structure of formula 101, formula 102, formula 103, or formula 104:
4. a mesylate salt of a derivative of any one of claims 1 to 3, having the formula II:
5. a process for the preparation of a derivative according to any one of claims 1 to 3, comprising the steps of:

reacting a compound with a structure shown in a formula III with a compound with a structure shown in a formula IV to obtain a compound with a structure shown in a formula V;

sequentially reducing and amidating the compound with the structure of the formula V to obtain a derivative with the structure of the formula I;

the R is₄、R₅、R₆、R₇、R₈、R₉、R₁₀、R₁₁、R₁₂、R₁₃、R₁₄、R₁₅、R₁₆、R₁₇、R₁₈、R₁₉And R₂₀Independently selected from hydrogen or deuterium;

in the formula III, M is F, Cl, Br or I.
6. The method of claim 5, wherein the compound having the structure of formula III is prepared by the following method:

reacting a compound with a structure shown in a formula VI with a compound with a structure shown in a formula VII to obtain a compound with a structure shown in a formula III;

the R is₁₁、R₁₂、R₁₃、R₁₄、R₁₅、R₁₆、R₁₇、R₁₈、R₁₉And R₂₀Independently selected from hydrogen or deuterium;

in the formula VI, R is F, Cl, Br or I.
7. The preparation method according to claim 6, wherein the reaction temperature of the compound with the structure of formula VI and the compound with the structure of formula VII is 90-100 ℃; the reaction time is 18-22 h.
8. The preparation method according to claim 5, wherein the reaction temperature of the compound with the structure of formula III and the compound with the structure of formula IV is 93-98 ℃; the reaction time is 5.5-6.5 h.
9. Use of a derivative according to any one of claims 1 to 3, a mesylate salt of a derivative according to any one of claims 4 to 6, or a derivative prepared by a method according to any one of claims 7 to 8 for the preparation of a medicament for the treatment of a tumor.
10. The use of claim 9, wherein the tumor is a malignant tumor.