CN109593064B

CN109593064B - Dithiocarbamate compounds as BTK inhibitors

Info

Publication number: CN109593064B
Application number: CN201811429773.6A
Authority: CN
Inventors: 尹玉新; 翟峥
Original assignee: Beijing Boyuan Precision Medical Technology Co ltd
Current assignee: Beijing Boyuan Precision Medical Technology Co., Ltd.
Priority date: 2018-11-28
Filing date: 2018-11-28
Publication date: 2020-08-11
Anticipated expiration: 2038-11-28
Also published as: CN109593064A

Abstract

The invention aims to provide a dithiocarbamate compound serving as a BTK inhibitor, a pharmaceutical composition, a preparation method and application thereof. The compound has a structure shown in the following general formula (I).

Description

Dithiocarbamate compounds as BTK inhibitors

Technical Field

The invention belongs to the technical field of medicinal chemistry, and relates to a novel compound, a method for preparing the compound, a medicinal composition and application thereof, in particular to a novel dithiocarbamic acid ester compound serving as a BTK inhibitor and application of the compound in preparing medicaments for treating or preventing cancers.

Background

Lymphoma is a relatively common tumor in our country, and ranks 11-13 of the causes of death of malignant tumors. B cell-derived malignancies include Chronic Lymphocytic Leukemia (CLL), Acute Lymphocytic Leukemia (ALL), Small Lymphocytic Lymphoma (SLL), Mantle Cell Lymphoma (MCL), Follicular Lymphoma (FL), Diffuse Large B Cell Lymphoma (DLBCL), Multiple Myeloma (MM), and Waldenstrom's Macroglobulinemia (WM). Currently, chemotherapy is the main treatment for lymphoma, and commonly used drugs include chlorambucil, cyclophosphamide, doxorubicin, vincristine, fludarabine, bendamustine and other cytotoxic drugs. CD20 positive B cell lymphomas can be combined with rituximab to improve treatment response rate and progression-free survival. However, chemotherapeutic drugs have limited antitumor efficacy and some patients have poor tolerance for adverse drug reactions. Therefore, the research and development of a new generation of molecular targeted antitumor drugs has become a hotspot, and the molecular targeted antitumor drugs have the advantages of specifically killing and killing tumor cells which grow rapidly, avoiding the damage to normal cells, and reducing the toxic and side effects of alopecia, gastrointestinal tract reaction, bone marrow inhibition and the like of the traditional chemotherapy drugs. Has wide prospect for target selection and drug development aiming at B cell malignant tumor.

Survival, proliferation, apoptosis and migration of B cell lymphoma cells depend on a tumor microenvironment, a BCR signal pathway is essential for survival of B cell-derived lymphoma cells, and a B cell receptor signal pathway is overactive in malignant B cells, so that normal differentiation and apoptosis of the B cells are inhibited, and abnormal proliferation is promoted. It is known that abnormal regulation of the BCR pathway occurs in malignancies of various B cell types, and thus, inhibition of activation of the BCR signaling pathway can ultimately prevent malignant B cell growth and proliferation. Bruton's Tyrosine Kinase (BTK) is a key regulator of the B Cell Receptor (BCR) signaling pathway, mainly expressed in B cells. The human BTK gene is located on the X chromosome (Xq21.33-Xq22) and includes 19 exons, with a full length of 37.5kb, and encodes a protein belonging to a member of the non-receptor tyrosine Tec family. The function of BTK is related to human immune diseases at the earliest, and in X-linked agammaglobulinemia (XLA), mutation of BTK is found to affect BTK protein expression, so that B cells are immature and dead, and the body of a patient is lack of antibodies and mature peripheral B lymphocytes and is immune. When an organism generates normal immune response, after BCR on the surface of a B cell is combined with an antigen of the B cell, the gathered BCR induces the activation of Src family kinases such as Lyn, Fyn and the like at the downstream, so that the migration of BTK to a plasma membrane and the autophosphorylation of a tyrosine 223 site are triggered, the activated BTK further induces the increase of the concentration of calcium ions in a cell, the activation of MAPKs and NF-kB signal pathways and the like through the activation of PLC-gamma, so that the expression of genes and cytokines is regulated, and the proliferation of the B cell is promoted. Continued activation of BTK leads to an abnormality in B cell function, can promote a variety of B cell malignancies, and elicit an autoimmune response. BTK therefore becomes an attractive target for the treatment of B-cell lymphoma and Rheumatoid Arthritis (RA).

In recent years small molecule inhibitors against BTK have evolved greatly in the treatment of hematologic malignancies and autoimmune diseases. BTK inhibitors currently used in preclinical and clinical studies are mainly divided into two classes, reversible BTK inhibitors and irreversible BTK inhibitors, wherein irreversible inhibitors include: ibrutinib (Ibrutinib) was the first BTK small molecule inhibitor to market, approved by the U.S. FDA in 11 months 2013 and 2 months 2014, for the treatment of Mantle Cell Lymphoma (MCL) and Chronic Lymphocytic Leukemia (CLL), respectively. The acarabretinib is a novel second-generation BTK inhibitor, improves the selectivity of protein tyrosine kinase, reduces side effects, is approved by FDA to be on the market in 2017 and 10 months, and is used for mantle cell lymphoma patients who have failed at least one-line treatment. Spebrutinib is a BTK inhibitor currently in clinical research and is used for treating chronic lymphocytic leukemia (clinic stage III) and rheumatoid arthritis (clinic stage II). The compounds are irreversible BTK inhibitors, the structure of which contains an acrylamide group, and the compounds can form covalent bonding with cysteine residue Cys481 of BTK protein, occupy the binding site of ATP and kinase to inhibit the activity of the kinase, and permanently destroy the biological activity of the protein. Another class of inhibitors are reversible inhibitors and compounds that have been introduced into clinical trials include GDC-0834 and BMS-986142, among others. The action mechanism of the compound is to competitively inhibit the combination of ATP and kinase, then the drug is metabolized and separated from the kinase, and dynamic equilibrium is achieved. Reversible inhibitors do not cause permanent damage to the target protein, but only temporarily interfere with its physiological function. In view of the hot direction of BTK inhibitor research, there is a need in the art to develop compounds with novel frameworks, which enhance the activity and selectivity of drugs, improve the toxic and side effects and physicochemical properties, and simultaneously cope with drug-resistant mutations caused by therapy.

The invention relates to a novel-structure aminodithioformate BTK inhibitor, and tests on kinase inhibition activity and cell proliferation inhibition activity of the BTK inhibitor prove that the BTK inhibitor has obvious effects on BTK kinase inhibition and tumor cell proliferation inhibition.

Disclosure of Invention

In the research process, the inventor finds a compound shown as a general formula (I) or a pharmaceutically acceptable salt thereof, wherein the compound has BTK inhibitory activity and can be used for treating BTK positive related cancers.

In a first aspect the present invention provides a compound having the general formula (i):

wherein:

l is selected from- (CH)₂)₂-、-(CH₂)₃-、-(CH₂)₄-; and/or

R₁、R₂The heterocyclic group is the same or different and is independently selected from hydrogen, alkyl, cycloalkyl, substituted or unsubstituted benzyl, substituted or unsubstituted heteroaryl alkyl, or forms a five-membered or six-membered heterocyclic group together with the N atom to which they are connected, wherein the heterocyclic group contains 1-2 heteroatoms selected from N, O or S; the heteroaryl group contains 1-3 heteroatoms selected from N, O or S; the substituted benzyl and substituted heteroarylalkyl groups having one or more substituents independently selected from C_1-4Alkoxy, cyano.

In a preferred embodiment of the first aspect of the invention L is selected from- (CH)₂)₂-、-(CH₂)₃-、-(CH₂)₄-; and/or

R₁、R₂Are the same or different and are each independently selected from hydrogen, methyl, ethyl, allyl, cyclopropyl, substituted or unsubstituted benzyl, picolyl, pyrazinylmethyl, or piperidinyl, morpholinyl, 2, 6-dimethylmorpholinyl, N-methylpiperazinyl, N-hydroxyethylpiperazinyl, N-acetylpiperazinyl, formed together with the N atom to which they are attached; the one or more substituents of said substituted benzyl group are independently selected from: methoxy group, cyano group.

According to an embodiment of the present invention, the compound of formula I according to the present invention may be at least one selected from the group consisting of:

2- [4- [ [4- [ (3-acrylamidophenyl) amine ] -5-fluoropyrimidin-2-yl ] amine ] phenoxyethyl methylaminodithioformate;

ethylaminodithiocarboxylic acid-2- [4- [ [4- [ (3-acrylamidophenyl) amine ] -5-fluoropyrimidin-2-yl ] amine ] phenoxyethyl ester;

2- [4- [ [4- [ (3-acrylamidophenyl) amine ] -5-fluoropyrimidin-2-yl ] amine ] phenoxyethyl allylaminodithioformate;

2- [4- [ [4- [ (3-acrylamidophenyl) amine ] -5-fluoropyrimidin-2-yl ] amine ] phenoxyethyl dimethylaminothioate;

diethylaminodithiocarboxylic acid-2- [4- [ [4- [ (3-acrylamidophenyl) amine ] -5-fluoropyrimidin-2-yl ] amine ] phenoxyethyl ester;

2- [4- [ [4- [ (3-acrylamidophenyl) amine ] -5-fluoropyrimidin-2-yl ] amine ] phenoxyethyl cyclopropylaminodithioformate;

piperidine-1-dithiocarboxylic acid-2- [4- [ [4- [ (3-acrylamidophenyl) amine ] -5-fluoropyrimidin-2-yl ] amine ] phenoxyethyl ester;

morpholine-4-dithiocarboxylic acid-2- [4- [ [4- [ (3-acrylamidophenyl) amine ] -5-fluoropyrimidin-2-yl ] amine ] phenoxyethyl ester;

2, 6-dimethylmorpholine-4-dithiocarboxylic acid 2- [4- [ [4- [ (3-acrylamidophenyl) amine ] -5-fluoropyrimidin-2-yl ] amine ] phenoxyethyl ester;

4-methylpiperazine-1-dithiocarboxylic acid-2- [4- [ [4- [ (3-acrylamidophenyl) amine ] -5-fluoropyrimidin-2-yl ] amine ] phenoxyethyl ester;

4-isopropylpiperazine-1-dithiocarboxylic acid-2- [4- [ [4- [ (3-acrylamidophenyl) amine ] -5-fluoropyrimidin-2-yl ] amine ] phenoxyethyl ester;

4- (2-hydroxyethyl) piperazine-1-dithiocarboxylic acid 2- [4- [ [4- [ (3-acrylamidophenyl) amine ] -5-fluoropyrimidin-2-yl ] amine ] phenoxyethyl ester;

4-acetylpiperazine-1-dithiocarboxylic acid-2- [4- [ [4- [ (3-acrylamidophenyl) amine ] -5-fluoropyrimidin-2-yl ] amine ] phenoxyethyl ester;

benzylamino dithiocarboxylic acid-2- [4- [ [4- [ (3-acrylamidophenyl) amine ] -5-fluoropyrimidin-2-yl ] amine ] phenoxyethyl ester;

4-methoxybenzyldithiocarbamic acid-2- [4- [ [4- [ (3-acrylamidophenyl) amine ] -5-fluoropyrimidin-2-yl ] amine ] phenoxyethyl ester;

3,4, 5-trimethoxybenzylamino dithiocarboxylic acid-2- [4- [ [4- [ (3-acrylamidophenyl) amine ] -5-fluoropyrimidin-2-yl ] amine ] phenoxyethyl ester;

4-cyanobenzylaminodithiocarboxylic acid-2- [4- [ [4- [ (3-acrylamidophenyl) amine ] -5-fluoropyrimidin-2-yl ] amine ] phenoxyethyl ester;

(pyridin-3-methyl) aminodithiocarboxylic acid-2- [4- [ [4- [ (3-acrylamidophenyl) amine ] -5-fluoropyrimidin-2-yl ] amine ] phenoxyethyl ester;

(pyridin-4-methyl) aminodithiocarboxylic acid-2- [4- [ [4- [ (3-acrylamidophenyl) amine ] -5-fluoropyrimidin-2-yl ] amine ] phenoxyethyl ester;

2- [4- [ [4- [ (3-acrylamidophenyl) amine ] -5-fluoropyrimidin-2-yl ] amine ] phenoxyethyl (pyridin-2-methyl) aminodithioformate;

2- [4- [ [4- [ (3-acrylamidophenyl) amine ] -5-fluoropyrimidin-2-yl ] amine ] phenoxyethyl (pyrazin-2-methyl) aminodithioformate;

3- [4- [ [4- [ (3-acrylamidophenyl) amine ] -5-fluoropyrimidin-2-yl ] amine ] phenoxypropyl dimethylaminothioate;

4-isopropylpiperazine-1-dithiocarboxylic acid-3- [4- [ [4- [ (3-acrylamidophenyl) amine ] -5-fluoropyrimidin-2-yl ] amine ] phenoxypropyl ester;

(pyridin-3-methyl) aminodithiocarboxylic acid-3- [4- [ [4- [ (3-acrylamidophenyl) amine ] -5-fluoropyrimidin-2-yl ] amine ] phenoxypropyl ester;

4- [4- [ [4- [ (3-acrylamidophenyl) amine ] -5-fluoropyrimidin-2-yl ] amine ] phenoxybutyl dimethylaminothioate;

(pyridin-3-methyl) aminodithiocarboxylic acid 4- [4- [ [4- [ (3-acrylamidophenyl) amine ] -5-fluoropyrimidin-2-yl ] amine ] phenoxybutyl ester;

as used herein, the term "pharmaceutically acceptable salts" refers to the conventional non-toxic salts formed by the reaction of a compound of formula I with an inorganic or organic acid. For example, the conventional non-toxic salts can be prepared by reacting a compound of formula I with an inorganic or organic acid. Wherein, the inorganic acid can be hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, sulfamic acid, phosphoric acid and the like, and the organic acid can be citric acid, tartaric acid, lactic acid, pyruvic acid, acetic acid, benzenesulfonic acid, p-toluenesulfonic acid, methanesulfonic acid, naphthalenesulfonic acid, ethanesulfonic acid, naphthalenedisulfonic acid, maleic acid, malic acid, malonic acid, fumaric acid, succinic acid, propionic acid, oxalic acid, trifluoroacetic acid, stearic acid, pamoic acid, hydroxymaleic acid, phenylacetic acid, benzoic acid, salicylic acid, glutamic acid, ascorbic acid, p-aminobenzenesulfonic acid, 2-acetoxybenzoic acid, isethionic acid and the like.

In a second aspect, the present invention provides a pharmaceutical composition comprising a compound according to the first aspect of the present invention or a pharmaceutically acceptable salt thereof.

A third aspect of the present invention provides the use of a compound as described above, or a pharmaceutically acceptable salt or solvate thereof, and a pharmaceutical composition for the manufacture of a medicament for modulating the catalytic activity of a tyrosine kinase, wherein the protein kinase is selected from bruton's tyrosine kinase.

In a fourth aspect, the present invention provides the use of the above-mentioned compounds or pharmaceutically acceptable salts thereof and pharmaceutical compositions for the manufacture of a medicament for the treatment or prevention of cancer.

In a preferred embodiment of the fourth aspect of the invention, the cancer is chronic lymphocytic leukemia, acute lymphocytic leukemia, small lymphocytic lymphoma, mantle cell lymphoma, follicular lymphoma, diffuse large B-cell lymphoma, multiple myeloma, waldenstrom's macroglobulinemia, and the like.

A fifth aspect of the invention provides a process for the preparation of a compound according to the first aspect of the invention:

(1) nucleophilic substitution reaction of 2, 4-dichloro-5-fluoropyrimidine and m-nitroaniline in isopropanol to obtain a compound 1, wherein the base of the reaction in the step is N, N-diisopropylethylamine;

(2) reacting p-nitrophenol with 1-bromo-2-chloroethane or 1-bromo-3-chloropropane or 1-bromo-3-chlorobutane under the action of potassium carbonate to obtain a compound 2, and reducing the compound 2 by iron powder and ammonium chloride to obtain a compound 3;

(3) carrying out nucleophilic substitution reaction on the compound 1 and the compound 3 to obtain a compound 4, reducing the compound 4 by iron powder and ammonium chloride to obtain a compound 5, and reacting the compound 5 with acryloyl chloride to obtain a compound 6;

(4) reacting the compound 6 with sodium iodide to obtain a compound 7, and reacting the compound 7 with carbon disulfide and different amines to obtain a target compound;

r described in the above synthetic scheme₁、R₂Are the same or different and are each independently selected from hydrogen, methyl, ethyl, allyl, cyclopropyl, substituted or unsubstituted benzyl, picolyl, pyrazinylmethyl, or piperidinyl, morpholinyl, 2, 6-dimethylmorpholinyl, N-methylpiperazinyl, N-hydroxyethylpiperazinyl, N-acetylpiperazinyl, formed together with the N atom to which they are attached; the one or more substituents of said substituted benzyl group are independently selected from: methoxy group, cyano group.

Detailed Description

The term "alkyl" as used herein refers to a group consisting of only carbon and hydrogen atoms, and having no unsaturation (e.g., double bonds, triple bonds, or rings), which encompasses a wide variety of possible geometric and stereoisomeric groups. This group is connected to the rest of the molecule by a single bond. By way of non-limiting examples of alkyl groups, mention may be made of the following linear or branched groups: methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl and seven further isomers thereof, n-hexyl and sixteen further isomers thereof, n-heptyl and various isomers thereof, n-octyl and various isomers thereof, n-nonyl and various isomers thereof, and n-decyl and various isomers thereof.

The term "cycloalkyl" as used herein refers to a saturated non-aromatic ring system consisting of at least 3 carbon atoms, which may be monocyclic, bicyclic, polycyclic, fused, bridged, or spiro. As non-limiting examples of cycloalkyl groups, the following groups may be cited: cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl; and fused, bridged or spiro ring groups formed from two or more of the above-mentioned monocyclic rings via a common side and a common carbon atom.

The term "alkoxy" as used herein refers to a group having an oxygen atom attached to the alkyl group and a single bond through the oxygen atom to the rest of the molecule, and encompasses a wide variety of possible geometric and stereoisomeric groups. By way of non-limiting examples of alkoxy radicals, mention may be made of the following linear or branched radicals: methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy, tert-butoxy, n-pentoxy and its other seven isomers, n-hexoxy and its other sixteen isomers, n-heptoxy and its various isomers, n-octoxy and its various isomers, n-nonoxy and its various isomers, n-decoxy and its various isomers.

The term "heteroaryl" as used herein refers to a 5-14 membered aromatic heterocyclic ring system having one or more heteroatoms independently selected from N, O or S, which may be monocyclic, bicyclic, polycyclic, wherein bicyclic and polycyclic rings may be formed from a single ring by single bond linkages or fused. As non-limiting examples of heteroaryl groups, the following groups may be cited: oxazolyl, isoxazolyl, imidazolyl, furyl, indolyl, isoindolyl, pyrrolyl, triazolyl, triazinyl, tetrazolyl, thienyl, thiazolyl, isothiazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, benzofuryl, benzothiazolyl, benzoxazolyl, benzimidazolyl, benzothienyl, benzopyranyl, carbazolyl, quinolyl, isoquinolyl, quinazolinyl, cinnolinyl, naphthyridinyl, pteridinyl, purinyl, quinoxalinyl, thiadiazolyl, indolizinyl, acridinyl, phenazinyl, phthalazinyl, coumarinyl, pyrazolopyridinyl, pyridopyridazinyl, pyrrolopyridyl, imidazopyridinyl, pyrazolopyridazinyl; and a group formed by the above-mentioned heteroaryl group by a single bond connection or a fusion connection.

The term "heteroarylalkyl" as used herein refers to an alkyl group wherein one or more hydrogen atoms are independently replaced by a heteroaryl group, wherein the heteroaryl and alkyl groups are as defined above.

The term "halogen" or "halo" as used herein refers to fluorine, chlorine, bromine or iodine.

The pharmaceutical composition of the present invention contains a compound according to the first aspect of the present invention or a pharmaceutically acceptable salt or solvate thereof as an active ingredient. In addition, the pharmaceutical composition may further comprise a pharmaceutically acceptable carrier, including but not limited to: water, salt solutions, alcohols, polyethylene glycols, polyhydroxyethoxylated castor oil, peanut oil, olive oil, gelatin, lactose, terra alba, sucrose, dextrin, magnesium carbonate, sugar, cyclodextrin, amylose, magnesium stearate, talc, gelatin, agar-agar, pectin, acacia, stearic acid or cellulose lower alkyl ethers, silicic acid, fatty acids, fatty acid amines, fatty acid monoglycerides and diglycerides, pentaerythritol fatty acid ethers, polyoxyethylene, hydroxymethylcellulose and polyvinylpyrrolidone. The pharmaceutical composition may further comprise one or more pharmaceutically acceptable adjuvants, wetting agents, emulsifying agents, suspending agents, preservatives, tonicity adjusting agents, buffering agents, sweetening agents, flavoring agents, coloring agents or any combination of the foregoing.

The pharmaceutical composition of the present invention can be formulated into any form of preparations, such as capsules, tablets, aerosols, solutions, suspensions, dragees, syrups, emulsions, ointments, pastes, injections, powders, granules, pastes, sustained-release preparations, foams. The drug of the present invention may be formulated into an oral administration preparation, a nasal administration preparation, a pulmonary administration preparation, an buccal preparation, a subcutaneous administration preparation, an intradermal administration preparation, a transdermal administration preparation, a parenteral administration preparation, a rectal administration preparation, a depot administration preparation, an intravenous administration preparation, an intraurethral administration preparation, an intramuscular administration preparation, an intranasal administration preparation, an ophthalmic administration preparation, an epidural administration preparation or a topical administration preparation, according to the administration route.

The "cancer" in the present invention includes various cancers known in the art, including but not limited to: chronic lymphocytic leukemia, acute lymphocytic leukemia, small lymphocytic lymphoma, mantle cell lymphoma, follicular lymphoma, diffuse large B-cell lymphoma, multiple myeloma, and waldenstrom's macroglobulinemia.

Examples

The present invention will be described in further detail with reference to examples, but the present invention is not limited to these examples.

In one exemplary embodiment, the compounds of the present invention are synthesized by the following method, wherein each substituent is as defined in formula (i).

Preparation example 1: 2-chloro-5-fluoro-N- (3-nitrophenyl) pyrimidin-4-amine

2, 4-dichloro-5-fluoropyrimidine (2g, 12mmol) and m-nitroaniline (1.38g, 10mmol) were dissolved in 20ml of isopropanol, DIEA (1.6ml,10mmol) was added, the mixture was heated under reflux at 90 ℃ for 6 hours until a large amount of yellow solid was formed, the mixture was cooled to room temperature, filtered by suction, the solid was washed with 95% ethanol and ether, and dried to give a pale yellow flocculent solid (1.77g, 66% yield).¹H NMR(400MHz,DMSO-d₆)10.38(s,1H),8.72(t,J＝2.1Hz,1H),8.41(d,J＝3.3Hz,1H),8.14(dd,J＝8.2,1.4Hz,1H),7.98–7.89(m,1H),7.65(t,J＝8.2Hz,1H)。

Preparative example 2 a: 1- (2-chloroethoxy) -4-nitrobenzene

P-nitrophenol (1.39g, 10mmol) and potassium carbonate (1.38g, 10mmol) were added to 25ml acetonitrile, 1-bromo-2-chloroethane (2.5ml, 30mmol) was added dropwise, and the reaction was refluxed at 80 ℃ overnight. After the reaction, water was added to dissolve potassium carbonate, extraction was performed with ethyl acetate, the organic phase was washed with saturated sodium bicarbonate and brine, and evaporated to dryness under reduced pressure to obtain a white solid (1.69g, yield 84%) which was used in the next reaction without purification.

Preparative example 2 b: 1- (3-chloropropoxy) -4-nitrobenzene

P-nitrophenol (1.39g, 10mmol) and potassium carbonate (1.38g, 10mmol) were added to 25ml acetonitrile, 1-bromo-3-chloropropane (1.5ml, 15mmol) was added dropwise, and the reaction was refluxed at 80 ℃ overnight. After the reaction, water was added to dissolve potassium carbonate, extraction was performed with ethyl acetate, the organic phase was washed with saturated sodium bicarbonate and brine, and evaporated to dryness under reduced pressure to obtain a white solid (1.92g, yield 89%), and the product was used in the next reaction without purification.

Preparative example 2 c: 1- (4-chlorobutoxy) -4-nitrobenzene

P-nitrophenol (1.39g, 10mmol) and potassium carbonate (1.38g, 10mmol) were added to 25ml acetonitrile, 1-bromo-4-chlorobutane (1.7ml, 15mmol) was added dropwise, and the reaction was refluxed at 80 ℃ overnight. After the reaction, water was added to dissolve potassium carbonate, extraction was performed with ethyl acetate, the organic phase was washed with saturated sodium bicarbonate and brine, and evaporated to dryness under reduced pressure to obtain a white solid (1.98g, yield 86%) which was used in the next reaction without purification.

Preparative example 3 a: 4- (2-chloroethoxy) aniline

The product of preparation example 2a (2g, 10mmol) was dissolved in 40ml of 75% ethanol, and reduced iron powder (2.8g, 50mmol) and ammonium chloride (0.8g, 15mmol) were added with stirring and the reaction was refluxed at 80 ℃ for about 1 h. The mixture was filtered under vacuum while hot, the filtrate was neutralized with saturated sodium bicarbonate, extracted with ethyl acetate, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and then separated with silica gel column (petroleum ether: ethyl acetate: 2:1), to obtain white crystals (1.25g, yield 73%).¹H NMR(400MHz,CDCl₃)6.79(d,J＝8.7Hz,2H),6.66(d,J＝8.6Hz,2H),4.17(t,J＝5.9Hz,2H),3.79(t,J＝5.9Hz,2H),3.45(s,2H).

Preparative example 3 b: 4- (3-chloropropoxy) aniline

Preparation of the product of example 2b (2.15g, 10)mmol) was dissolved in 30ml of 75% ethanol, and reduced iron powder (2.8g, 50mmol) and ammonium chloride (0.8g, 15mmol) were added with stirring, and the reaction was refluxed at 80 ℃ for about 1 h. Vacuum-filtering while hot, adding saturated sodium bicarbonate to the filtrate for neutralization, extracting with ethyl acetate, washing the organic phase with saturated sodium chloride, drying over anhydrous sodium sulfate, separating with silica gel column (petroleum ether: ethyl acetate 4:1), and purifying to obtain white crystals 1-c (1.71g, yield 92%).¹H NMR(400MHz,CDCl₃)6.74(d,J＝8.7Hz,2H),6.66(d,J＝8.6Hz,2H),4.06(t,J＝6.3Hz,2H),3.68(t,J＝6.3Hz,2H),3.46(s,2H),1.98-1.83(m,2H)。

Preparative example 3 c: 4- (4-chloropropoxy) aniline

The procedure described for preparation example 3a was followed except that the starting material preparation example 2a product was changed to the preparation example 2c product.

Preparative example 4 a: n is a radical of²- [4- (2-chloroethoxy) phenyl group]-5-fluoro-N⁴- (3-nitrophenyl) pyrimidine-2, 4-diamine

The product of preparation example 3a (940mg, 5.5mmol) and the product of preparation example 1 (1.34g, 5mmol) were dissolved in 25ml of isopropanol, trifluoroacetic acid (0.5ml, 6mmol) was added dropwise, heating and refluxing were carried out at 90 ℃ for 8h, cooling to room temperature to precipitate a white solid, suction filtration was carried out, the cake was washed with 95% ethanol, and drying was carried out to obtain a white solid powder (1.6g, yield 79%) which was used in the next reaction without purification.¹H NMR(400MHz,DMSO-d₆)10.73(s,1H),10.25(s,1H),8.51(t,J＝2.0Hz,1H),8.36(d,J＝4.7Hz,1H),8.20(d,J＝7.5Hz,1H),7.98(dd,J＝8.2,1.7Hz,1H),7.62(t,J＝8.2Hz,1H),7.39(d,J＝8.9Hz,2H),6.87(d,J＝9.0Hz,2H),4.24–4.18(m,2H),3.97–3.91(m,2H)。

Preparative example 4 b: n is a radical of²- [4- (3-Chloropropoxy) phenyl group]-5-fluoro-N⁴- (3-nitrophenyl) pyrimidine-2, 4-diamine

The procedure described for preparation example 4a was followed except that the starting material preparation example 3a was changed to the preparation example 3 b.

Preparative example 4 c: n is a radical of²- [4- (4-Neobutoxy) phenyl group]-5-fluoro-N⁴- (3-nitrophenyl) pyrimidine-2, 4-diamine

The procedure described for preparation example 4a was followed except that the starting material preparation example 3a was changed to the preparation example 3 c.

Preparative example 5 a: n is a radical of⁴- (3-aminophenyl) -N²- [4- (2-chloroethoxy) phenyl group]-5-fluoropyrimidine-2, 4-diamine

Preparation example (807mg, 2mmol) was dissolved in 20ml of 75% ethanol, and reduced iron powder (450mg, 10mmol) and ammonium chloride (160mg, 3mmol) were added with stirring, and the reaction was refluxed at 80 ℃ for about 2 hours. The reaction mixture was filtered while hot, the filtrate was extracted with ethyl acetate, the organic phase was washed with saturated brine, and evaporated to dryness under reduced pressure to give a white solid (690mg, yield 93%), which was used directly in the next reaction without purification.

Preparative example 5 b: n is a radical of⁴- (3-aminophenyl) -N²- [4- (3-Chloropropoxy) phenyl group]-5-fluoropyrimidine-2, 4-diamine

The procedure described for preparation example 5a was followed except that the starting material preparation example 4a was changed to the preparation example 4 b.

Preparative example 5 c: n is a radical of⁴- (3-aminophenyl) -N²- [4- (4-Neobutoxy) phenyl group]-5-fluoropyrimidines2, 4-diamines

The procedure described for preparation example 5a was followed except that the starting material preparation example 4a was changed to the preparation example 4 c.

Preparative example 6 a: n-3- [ [2- [ [4- (2-chloroethoxy) phenyl ] amino-5-fluoropyrimidin-4-yl ] amino ] phenyl acrylamide

The product of preparation example 5a (374mg, 1mmol) was dissolved in 10ml DMF, sodium carbonate (212mg, 1mmol) was added, the reaction was stirred in an ice bath, acryloyl chloride (85 μ L, 1.05mmol) was slowly added dropwise, after all addition, the reaction was reacted for 0.5h, ethyl acetate and water were added to the reaction solution, ethyl acetate was extracted, saturated brine was washed, dried over anhydrous sodium sulfate, and separated by silica gel column (petroleum ether: ethyl acetate ═ 1:1) to obtain a white powdery solid 1-h (372mg, yield 87%).¹H NMR(400MHz,DMSO-d₆)10.13(s,1H),9.39(s,1H),9.01(s,1H),8.08(d,J＝3.6Hz,1H),7.96(s,1H),7.57(d,J＝8.8Hz,2H),7.51(d,J＝7.9Hz,1H),7.43(d,J＝7.9Hz,1H),7.30(t,J＝8.1Hz,1H),6.80(d,J＝8.8Hz,2H),6.48(dd,J＝16.9,10.1Hz,1H),6.28(d,J＝16.9Hz,1H),5.77(d,J＝10.2Hz,1H),4.21–4.13(m,2H),3.95–3.88(m,2H)。

Preparative example 6 b: n-3- [ [2- [ [4- (3-chloropropoxy) phenyl ] amino-5-fluoropyrimidin-4-yl ] amino ] phenyl acrylamide

The procedure described for preparation example 6a was followed except that the starting material preparation example 5a was changed to the preparation example 5 b.

Preparative example 6 c: n-3- [ [2- [ [4- (4-chlorobutoxy) phenyl ] amino-5-fluoropyrimidin-4-yl ] amino ] phenyl acrylamide

The procedure described for preparation example 6a was followed except that the starting material preparation example 5a was changed to the preparation example 5 c.

Preparative example 7 a: n-3- [ [ 5-fluoro-2- [ [4- (2-iodoethoxy) phenyl ] amino ] pyrimidin-4-yl ] amino ] phenylacrylamide

The product of preparation example 6a (428mg, 1mmol) and sodium iodide (750mg, 5mmol) were dissolved in 10ml acetone and reacted at 60 ℃ under reflux for 48h, with sodium chloride precipitate being formed continuously during the reaction. After completion of the reaction, water was added, and the mixture was extracted with ethyl acetate, washed with saturated brine, and evaporated to dryness under reduced pressure to obtain a white solid powder (478mg, yield 92%).

Preparative example 7 b: n-3- [ [ 5-fluoro-2- [ [4- (3-iodopropoxy) phenyl ] amino ] pyrimidin-4-yl ] amino ] phenylacrylamide

The procedure described for preparation 7a was followed except that the starting material, preparation 6a, was changed to the preparation 6 b.

Preparative example 7 c: n-3- [ [ 5-fluoro-2- [ [4- (4-iodobutyloxy) phenyl ] amino ] pyrimidin-4-yl ] amino ] phenylacrylamide

The procedure described for preparation 7a was followed except that the starting material, preparation 6a, was changed to the preparation 6 c.

Example 1: methyldithiocarbamic acid-2- [4- [ [4- [ (3-acrylamidophenyl) amine ] -5-fluoropyrimidin-2-yl ] amine ] phenoxyethyl ester

Methylamine hydrochloride (81mg,1.2mmol) was dissolved in 15mL acetone, triethylamine (556. mu.L, 4mmol) and carbon disulfide (120. mu.L, 2mmol) were added dropwise, and the mixture was stirred at room temperature for 30 min. Then, the product of preparation example 7a (519mg,1mmol) was added and reacted at 60 ℃ for 1 h. To the reaction system, silica gel was directly added, the solvent was dried under reduced pressure, and the mixture was separated with silica gel (P: E ═ 1:1) to obtain a white solid (245mg, yield 49%).¹H NMR(400MHz,DMSO-d₆)10.11(s,1H),10.02(d,J＝3.5Hz,1H),9.36(s,1H),8.97(s,1H),8.07(d,J＝3.3Hz,1H),7.96(s,1H),7.55(d,J＝8.7Hz,2H),7.50(d,J＝7.8Hz,1H),7.42(d,J＝7.7Hz,1H),7.29(t,J＝8.0Hz,1H),6.78(d,J＝8.7Hz,2H),6.47(dd,J＝16.9,10.1Hz,1H),6.27(d,J＝16.8Hz,1H),5.75(d,J＝11.2Hz,1H),4.07(t,J＝6.3Hz,2H),3.57(t,J＝6.2Hz,2H),3.06(d,J＝4.2Hz,3H)。¹³C NMR(101MHz,DMSO-d₆)196.37,163.59,156.16(d,J＝2.5Hz),153.13,150.25(d,J＝10.7Hz),142.07,141.25(d,J＝19.0Hz),139.62,139.53(d,J＝6.4Hz),134.85,132.38,129.10,127.31,120.73,117.84,115.32,114.78,113.94,66.97,34.29,33.85。HRMS(ESI)m/z calcd forC23H24FN6O2S2[M+H]+499.13807.Found:499.13693。

Example 2: ethylaminodithiocarboxylic acid-2- [4- [ [4- [ (3-acrylamidophenyl) amine ] -5-fluoropyrimidin-2-yl ] amine ] phenoxyethyl ester

The procedure described in preparation example 1 was followed, except that the starting methylamine hydrochloride was changed to ethylamine in 52% yield.¹HNMR(400MHz,DMSO-d₆)10.11(s,1H),10.04(t,J＝4.9Hz,1H),9.37(s,1H),8.97(s,1H),8.06(d,J＝3.7Hz,1H),7.94(s,1H),7.54(d,J＝9.0Hz,2H),7.49(d,J＝8.0Hz,1H),7.41(d,J＝8.1Hz,1H),7.28(t,J＝8.1Hz,1H),6.78(d,J＝9.0Hz,2H),6.46(dd,J＝17.0,10.1Hz,1H),6.26(dd,J＝17.0,1.9Hz,1H),5.75(dd,J＝10.1,1.9Hz,1H),4.06(t,J＝6.5Hz,2H),3.62-3.55(m,4H),1.14(t,J＝7.3Hz,3H)。¹³C NMR(101MHz,DMSO-d₆)195.42,163.57,156.12(d,J＝2.6Hz),153.13,150.25(d,J＝10.7Hz),142.05,141.19(d,J＝19.0Hz),139.60,139.52(d,J＝4.6Hz),134.81,132.38,129.11,127.32,120.73,117.84,115.31,114.76,113.92,66.98,42.28,33.65,13.54。HRMS(ESI)m/z calcd forC₂₄H₂₆FN₆O₂S₂[M+H]⁺513.15372.Found:513.15381。

Example 3: 2- [4- [ [4- [ (3-acrylamidophenyl) amine ] -5-fluoropyrimidin-2-yl ] amine ] phenoxyethyl allylaminodithioformate

The procedure described in preparation example 1 was followed, except that the starting methylamine hydrochloride was changed to allylamine, yield 43%.¹HNMR(400MHz,DMSO-d₆)10.24(t,J＝5.2Hz,1H),10.12(s,1H),9.38(s,1H),8.98(s,1H),8.08(d,J＝3.7Hz,1H),7.96(s,1H),7.55(d,J＝9.0Hz,2H),7.51(d,J＝8.0Hz,1H),7.43(d,J＝8.0Hz,1H),7.29(t,J＝8.1Hz,1H),6.79(d,J＝9.0Hz,2H),6.48(dd,J＝17.0,10.1Hz,1H),6.28(dd,J＝17.0,1.9Hz,1H),5.88(ddd,J＝22.7,10.6,5.6Hz,1H),5.76(dd,J＝10.1,1.9Hz,1H),5.18(ddd,J＝13.7,11.7,1.5Hz,2H),4.26(t,J＝5.4Hz,2H),4.08(t,J＝6.5Hz,2H),3.59(t,J＝6.4Hz,2H)。¹³C NMR(101MHz,DMSO-d₆)196.43,163.57,156.13(d,J＝2.6Hz),153.11,150.24(d,J＝10.7Hz),142.06,141.23(d,J＝20.6Hz),139.61,139.52(d,J＝5.4Hz),134.84,133.29,132.38,129.11,127.33,120.71,117.84,117.47,115.31,114.76,113.93,66.94,49.42,33.83。HRMS(ESI)m/z calcd for C₂₅H₂₆FN₆O₂S₂[M+H]⁺525.15372.Found:525.15430。

Example 4: dimethyldithiocarbamate-2- [4- [ [4- [ (3-acrylamidophenyl) amine ] -5-fluoropyrimidin-2-yl ] amine ] phenoxyethyl ester

The procedure described in preparation example 1 was followed except that the starting methylamine salt was usedThe acid salt was changed to dimethylamine hydrochloride, yield 42%.¹H NMR(400MHz,DMSO-d₆)10.12(s,1H),9.38(s,1H),8.98(s,1H),8.07(d,J＝3.6Hz,1H),7.95(s,1H),7.54(d,J＝8.9Hz,2H),7.50(d,J＝8.3Hz,1H),7.42(d,J＝8.0Hz,1H),7.28(t,J＝8.1Hz,1H),6.78(d,J＝8.9Hz,2H),6.47(dd,J＝16.9,10.1Hz,1H),6.27(dd,J＝16.9,1.5Hz,1H),5.78–5.72(m,1H),4.10(t,J＝6.3Hz,2H),3.62(t,J＝6.3Hz,2H),3.48(s,3H),3.37(s,3H)。¹³C NMR(101MHz,DMSO-d₆)195.17,163.57,156.12(d,J＝2.7Hz),153.10,150.24(d,J＝11.0Hz),142.05,141.21(d,J＝20.4Hz),139.60,139.51(d,J＝5.0Hz),134.83,132.37,129.12,127.33,120.72,117.83,115.29,114.76,113.90,66.48,45.63,41.79,36.34。HRMS(ESI)m/z calcdfor C₂₄H₂₆FN₆O₂S₂[M+H]⁺513.15372.Found:513.15308。

Example 5: diethylaminodithiocarboxylic acid-2- [4- [ [4- [ (3-acrylamidophenyl) amine ] -5-fluoropyrimidin-2-yl ] amine ] phenoxyethyl ester

The procedure described in preparation example 1 was followed except that the starting methylamine hydrochloride was changed to diethylamine in a yield of 46%.¹HNMR(400MHz,DMSO-d₆)10.12(s,1H),9.37(s,1H),8.98(s,1H),8.07(d,J＝3.7Hz,1H),7.96(s,1H),7.55(d,J＝9.0Hz,2H),7.51(d,J＝8.0Hz,1H),7.42(d,J＝8.1Hz,1H),7.29(t,J＝8.1Hz,1H),6.80(d,J＝9.0Hz,2H),6.47(dd,J＝17.0,10.1Hz,1H),6.27(dd,J＝17.0,1.9Hz,1H),5.75(dd,J＝10.1,1.9Hz,1H),4.11(t,J＝6.4Hz,2H),3.98(q,J＝7.0Hz,2H),3.77(q,J＝7.0Hz,2H),3.63(t,J＝6.4Hz,2H),1.27–1.16(m,6H)。¹³C NMR(101MHz,DMSO-d₆)193.88,163.56,156.15(d,J＝2.8Hz),153.11,150.23(d,J＝10.6Hz),142.06,141.26(d,J＝19.6Hz),139.61,139.52(d,J＝6.2Hz),134.86,132.39,129.10,127.27,120.70,117.81,115.28,114.79,113.90,66.55,49.75,47.07,35.87,12.84,11.82。HRMS(ESI)m/z calcd for C₂₆H₃₀FN₆O₂S₂[M+H]⁺541.18502.Found:541.18500。

Example 6: cyclopropanedithiocarbamic acid 2- [4- [ [4- [ (3-acrylamidophenyl) amine ] -5-fluoropyrimidin-2-yl ] amine ] phenoxyethyl ester

The procedure described in preparative example 1 was followed except that the starting methylamine hydrochloride was changed to cyclopropylamine, yield 37%.¹HNMR(400MHz,DMSO-d₆)10.13(s,1H),10.07(s,1H),9.38(s,1H),8.99(s,1H),8.08(s,1H),7.95(s,1H),7.55(d,J＝6.1Hz,2H),7.50(d,J＝6.4Hz,1H),7.42(d,J＝6.3Hz,1H),7.30(d,J＝7.2Hz,1H),6.79(s,2H),6.47(dd,J＝15.9,10.4Hz,1H),6.27(d,J＝16.6Hz,1H),5.76(d,J＝5.3Hz,1H),4.09(t,J＝6.3Hz,2H),3.60(t,J＝6.3Hz,2H),3.43-3.31(m,1H),0.73(dd,J＝40.8,13.5Hz,4H)。¹³C NMR(101MHz,DMSO-d₆)196.78,163.56,156.12,153.09,150.22(d,J＝9.7Hz),142.04,141.26(d,J＝19.5Hz),139.58,139.51(d,J＝5.5Hz),134.83,132.37,129.11,127.34,120.68,117.81,115.28,114.74,113.89,66.90,33.76,30.58,7.87,6.60。HRMS(ESI)m/z calcd forC₂₅H₂₆FN₆O₂S₂[M+H]⁺525.15372.Found:525.15295。

Example 7: piperidine-1-dithiocarboxylic acid-2- [4- [ [4- [ (3-acrylamidophenyl) amine ] -5-fluoropyrimidin-2-yl ] amine ] phenoxyethyl ester

The procedure described in preparation example 1 was followed except that the starting methylamine hydrochloride was changed to piperidine hydrochloride in a 36% yield.¹HNMR(400MHz,DMSO-d₆)10.12(s,1H),9.38(s,1H),8.99(s,1H),8.07(d,J＝3.7Hz,1H),7.96(s,1H),7.55(d,J＝9.0Hz,2H),7.51(d,J＝8.0Hz,1H),7.42(d,J＝8.1Hz,1H),7.29(t,J＝8.1Hz,1H),6.79(d,J＝9.0Hz,2H),6.47(dd,J＝17.0,10.1Hz,1H),6.27(dd,J＝17.0,1.9Hz,1H),5.75(dd,J＝10.1,2.0Hz,1H),4.23(s,2H),4.11(t,J＝6.4Hz,2H),3.91(d,J＝4.9Hz,2H),3.65(t,J＝6.4Hz,2H),1.61(d,J＝4.5Hz,6H)。¹³C NMR(101MHz,DMSO-d₆)193.69,163.57,156.12(d,J＝2.8Hz),153.13,150.25(d,J＝10.8Hz),142.06,141.19(d,J＝18.9Hz),139.61,139.52(d,J＝4.7Hz),134.83,132.39,129.10,127.29,120.73,117.83,115.29,114.78,113.91,66.61,53.07,51.44,35.93,26.26,25.67,24.01。HRMS(ESI)m/z calcd for C₂₇H₃₀FN₆O₂S₂[M+H]⁺553.18502.Found:553.18414。

Example 8: morpholine-4-dithiocarboxylic acid-2- [4- [ [4- [ (3-acrylamidophenyl) amine ] -5-fluoropyrimidin-2-yl ] amine ] phenoxyethyl ester

The procedure described in preparation example 1 was followed except that the starting methylamine hydrochloride was changed to morpholine in a yield of 60%.¹HNMR(400MHz,DMSO-d₆)10.07(s,1H),9.32(s,1H),8.93(s,1H),8.01(d,J＝3.6Hz,1H),7.90(s,1H),7.49(d,J＝8.9Hz,2H),7.45(d,J＝8.1Hz,1H),7.37(d,J＝8.0Hz,1H),7.23(t,J＝8.1Hz,1H),6.73(d,J＝8.9Hz,2H),6.42(dd,J＝16.9,10.1Hz,1H),6.22(d,J＝16.9Hz,1H),5.70(d,J＝10.1Hz,1H),4.19(s,2H),4.07(t,J＝6.3Hz,2H),3.90(s,2H),3.62(s,6H)。¹³C NMR(101MHz,DMSO-d₆)195.58,163.52,156.07(d,J＝2.6Hz),153.02,150.18(d,J＝10.9Hz),142.00,141.17(d,J＝20.2Hz),139.55,139.47(d,J＝5.0Hz),134.81,132.32,129.06,127.28,120.66,117.77,115.23,114.71,113.83,66.39,66.01,65.98,51.78,50.68,35.74。HRMS(ESI)m/z calcd forC₂₆H₂₈FN₆O₃S₂[M+H]⁺555.16428.Found:555.16376。

Example 9: 2, 6-Dimethylmorpholine-4-dithiocarboxylic acid 2- [4- [ [4- [ (3-acrylamidophenyl) amine ] -5-fluoropyrimidin-2-yl ] amine ] phenoxyethyl ester

The procedure described in preparation example 1 was followed except that the starting methylamine hydrochloride was changed2, 6-dimethylmorpholine in 43% yield.¹H NMR(400MHz,DMSO-d₆)10.12(s,1H),9.38(s,1H),8.98(s,1H),8.07(d,J＝3.7Hz,1H),7.95(t,J＝1.7Hz,1H),7.55(d,J＝9.1Hz,2H),7.51(d,J＝8.2Hz,1H),7.43(d,J＝8.6Hz,1H),7.29(t,J＝8.1Hz,1H),6.79(d,J＝9.1Hz,2H),6.47(dd,J＝17.0,10.1Hz,1H),6.27(dd,J＝17.0,2.0Hz,1H),5.76(dd,J＝10.1,2.0Hz,1H),5.28(s,1H),4.43(s,1H),4.14(t,J＝6.3Hz,2H),3.68(t,J＝6.3Hz,2H),3.63-3.55(m,2H),3.17–2.69(m,2H),1.15(d,J＝6.2Hz,6H)。¹³CNMR(101MHz,DMSO-d₆)195.41,163.56,156.14(d,J＝2.8Hz),153.06,150.23(d,J＝10.8Hz),142.06,141.25(d,J＝20.2Hz),139.62,139.53(d,J＝5.9Hz),134.89,132.39,129.10,127.30,120.70,117.82,115.28,114.77,113.89,71.24,66.45,56.38,55.21,35.89,18.81。HRMS(ESI)m/z calcd for C₂₈H₃₂FN₆O₃S₂[M+H]⁺583.19558.Found:583.19489。

Example 10: 4-methylpiperazine-1-dithiocarboxylic acid-2- [4- [ [4- [ (3-acrylamidophenyl) amine ] -5-fluoropyrimidin-2-yl ] amine ] phenoxyethyl ester

The procedure described in preparation example 1 was followed except that the starting methylamine hydrochloride was changed to N-methylpiperazine, yield 52%.¹HNMR(400MHz,DMSO-d₆)10.12(s,1H),9.37(s,1H),8.98(s,1H),8.07(d,J＝3.7Hz,1H),7.96(s,1H),7.55(d,J＝9.0Hz,2H),7.51(d,J＝8.0Hz,1H),7.42(d,J＝8.2Hz,1H),7.28(t,J＝8.1Hz,1H),6.79(d,J＝9.0Hz,2H),6.47(dd,J＝17.0,10.1Hz,1H),6.27(dd,J＝17.0,1.9Hz,1H),5.75(dd,J＝10.1,1.9Hz,1H),4.24(s,2H),4.12(t,J＝6.3Hz,2H),3.92(s,2H),3.66(t,J＝6.3Hz,2H),2.44–2.33(m,4H),2.20(s,3H)。¹³C NMR(101MHz,DMSODMSO-d₆1,163.57,156.14(d,J＝2.6Hz),153.08,150.23(d,J＝10.7Hz),142.07,141.26(d,J＝19.4Hz),139.62,139.53(d,J＝5.8Hz),134.88,132.40,129.11,127.31,120.69,117.81,115.27,114.77,113.88,66.50,54.42,51.52,50.00,45.51,35.94。HRMS(ESI)m/zcalcd for C₂₇H₃₁FN₇O₂S₂[M+H]⁺568.19592.Found:568.19537。

Example 11: 4-isopropylpiperazine-1-dithiocarboxylic acid 2- [4- [ [4- [ (3-acrylamidophenyl) amine ] -5-fluoropyrimidin-2-yl ] amine ] phenoxyethyl ester

The procedure described in preparation example 1 was followed except that the starting methylamine hydrochloride was changed to N-isopropylpiperazine with a yield of 63%.¹H NMR(400MHz,DMSO-d₆)10.12(s,1H),9.37(s,1H),8.98(s,1H),8.07(d,J＝3.7Hz,1H),7.95(s,1H),7.55(d,J＝9.1Hz,2H),7.52(d,J＝8.3Hz,1H),7.42(d,J＝8.7Hz,1H),7.29(t,J＝8.1Hz,1H),6.79(d,J＝9.1Hz,2H),6.47(dd,J＝17.0,10.1Hz,1H),6.27(dd,J＝17.0,2.0Hz,1H),5.76(dd,J＝10.1,2.0Hz,1H),4.23(s,2H),4.12(t,J＝6.4Hz,2H),3.91(s,2H),3.66(t,J＝6.3Hz,2H),2.69(hept,J＝6.4Hz,1H),2.51(dd,J＝5.4,3.6Hz,4H),0.97(d,J＝6.5Hz,6H)。¹³C NMR(101MHz,DMSO-d₆)194.72,163.56,156.14(d,J＝2.8Hz),153.09,150.23(d,J＝10.8Hz),142.06,141.25(d,J＝19.2Hz),139.61,139.53(d,J＝6.1Hz),134.87,132.39,129.10,127.29,120.69,117.81,115.27,114.78,113.88,66.53,53.94,52.07,50.62,48.21,35.87,18.57。HRMS(ESI)m/z calcd for C₂₉H₃₅FN₇O₂S₂[M+H]⁺596.22722.Found:596.22754。

Example 12: 4- (2-hydroxyethyl) piperazine-1-dithiocarboxylic acid 2- [4- [ [4- [ (3-acrylamidophenyl) amine ] -5-fluoropyrimidin-2-yl ] amine ] phenoxyethyl ester

The procedure described in preparation example 1 was followed except that the starting methylamine hydrochloride was changed to 1- (2-hydroxyethyl) piperazine in a yield of 32%.¹H NMR(400MHz,DMSO-d₆)10.13(s,1H),9.38(s,1H),8.99(s,1H),8.07(d,J＝3.3Hz,1H),7.96(s,1H),7.55(d,J＝8.7Hz,2H),7.51(d,J＝7.8Hz,1H),7.42(d,J＝7.8Hz,1H),7.29(t,J＝8.0Hz,1H),6.79(d,J＝8.7Hz,2H),6.47(dd,J＝16.9,10.1Hz,1H),6.27(d,J＝16.9Hz,1H),5.76(d,J＝10.1Hz,1H),4.49(s,1H),4.25(s,2H),4.12(t,J＝6.1Hz,2H),3.93(s,2H),3.66(t,J＝6.0Hz,2H),3.53(s,2H),2.51(t,J＝7.9Hz,6H),2.44(t,J＝5.9Hz,2H)。¹³C NMR(101MHz,DMSO-d₆)194.88,163.56,156.13(d,J＝2.6Hz),153.07,150.22(d,J＝10.8Hz),142.05,141.25(d,J＝18.4Hz),139.61,139.52(d,J＝5.7Hz),134.86,132.38,129.11,127.32,120.68,117.80,115.27,114.76,113.88,66.50,59.93,58.94,52.98,51.64,50.29,35.90。HRMS(ESI)m/zcalcd for C₂₈H₃₃FN₇O₃S₂[M+H]⁺598.20648.Found:598.20544。

Example 13: 4-Acetylpiperazine-1-dithiocarboxylic acid-2- [4- [ [4- [ (3-acrylamidophenyl) amine ] -5-fluoropyrimidin-2-yl ] amine ] phenoxyethyl ester

The procedure described in preparation example 1 was followed except that the starting methylamine hydrochloride was changed to N-acetylpiperazine with a yield of 57%.¹H NMR(400MHz,DMSO-d₆)10.13(s,1H),9.39(s,1H),9.00(s,1H),8.08(d,J＝3.7Hz,1H),7.95(s,1H),7.55(d,J＝9.1Hz,2H),7.51(d,J＝8.1Hz,1H),7.42(d,J＝8.6Hz,1H),7.29(t,J＝8.1Hz,1H),6.79(d,J＝9.1Hz,2H),6.47(dd,J＝17.0,10.1Hz,1H),6.27(dd,J＝17.0,2.0Hz,1H),5.76(dd,J＝10.1,2.0Hz,1H),4.44-4.18(m,2H),4.14(t,J＝6.3Hz,2H),4.09–3.91(m,2H),3.68(t,J＝6.3Hz,2H),3.60(dd,J＝10.8,7.2Hz,4H),2.03(s,3H)。¹³CNMR(101MHz,DMSO-d₆)195.56,169.14,163.56,156.11(d,J＝2.8Hz),153.07,150.24(d,J＝10.8Hz),142.05,141.20(d,J＝20.5Hz),139.60,139.52(d,J＝4.4Hz),134.86,132.38,129.11,127.34,120.71,117.83,115.28,114.76,113.88,66.42,51.07,49.72,44.90,35.93,21.69。HRMS(ESI)m/zcalcd for C₂₈H₃₁FN₇O₃S₂[M+H]⁺596.19083.Found:596.19122。

Example 14: benzylaminodithioic acid 2- [4- [ [4- [ (3-acrylamidophenyl) amine ] -5-fluoropyrimidin-2-yl ] amine ] phenoxyethyl ester

The procedure described in preparation example 1 was followed except that the starting methylamine hydrochloride was changed to benzylamine hydrochloride in 49% yield.¹HNMR(400MHz,DMSO-d₆)10.56(t,J＝5.6Hz,1H),10.13(s,1H),9.38(s,1H),8.99(s,1H),8.08(d,J＝3.7Hz,1H),7.96(s,1H),7.56(d,J＝9.0Hz,2H),7.51(d,J＝8.1Hz,1H),7.43(d,J＝8.1Hz,1H),7.38–7.25(m,6H),6.79(d,J＝9.0Hz,2H),6.48(dd,J＝17.0,10.1Hz,1H),6.27(dd,J＝17.0,1.9Hz,1H),5.76(dd,J＝10.1,1.9Hz,1H),4.88(d,J＝5.6Hz,2H),4.10(t,J＝6.4Hz,2H),3.61(t,J＝6.4Hz,2H)。¹³C NMR(101MHz,DMSO-d₆)196.89,163.57,156.14(d,J＝2.8Hz),153.11,150.24(d,J＝10.8Hz),142.06,141.25(d,J＝19.1Hz),139.61,139.52(d,J＝5.6Hz),137.73,134.86,132.38,129.11,128.85,128.16,127.69,127.33,120.70,117.83,115.30,114.77,113.93,66.93,50.20,33.92。HRMS(ESI)m/z calcd for C₂₉H₂₈FN₆O₂S₂[M+H]⁺575.16937.Found:575.16913。

Example 15: 4-Methoxybenzyldithiocarbamic acid 2- [4- [ [4- [ (3-acrylamidophenyl) amine ] -5-fluoropyrimidin-2-yl ] amine ] phenoxyethyl ester

The procedure described in preparation example 1 was followed except that the starting methylamine hydrochloride was changed to p-methylbenzylamine, yield 61%.¹HNMR(400MHz,DMSO-d₆)10.49(t,J＝5.5Hz,1H),10.13(s,1H),9.39(s,1H),9.00(s,1H),8.08(d,J＝3.7Hz,1H),7.96(s,1H),7.55(d,J＝9.0Hz,2H),7.51(d,J＝8.0Hz,1H),7.43(d,J＝8.1Hz,1H),7.31–7.23(m,3H),6.90(d,J＝8.6Hz,2H),6.78(d,J＝9.0Hz,2H),6.47(dd,J＝17.0,10.1Hz,1H),6.27(dd,J＝17.0,1.9Hz,1H),5.78–5.71(m,1H),4.79(d,J＝5.5Hz,2H),4.08(t,J＝6.4Hz,2H),3.73(s,3H),3.60(t,J＝6.4Hz,2H)。¹³C NMR(101MHz,DMSO-d₆)196.38,163.57,159.01,156.13(d,J＝2.6Hz),153.09,150.23(d,J＝10.7Hz),142.05,141.25(d,J＝18.9Hz),139.60,139.52(d,J＝5.6Hz),134.84,132.36,129.69,129.62,129.11,127.35,120.67,117.81,115.28,114.74,114.24,113.89,66.92,55.54,49.77,33.84。HRMS(ESI)m/z calcdfor C₃₀H₃₀FN₆O₃S₂[M+H]⁺605.17993.Found:605.17920。

Example 16: 3,4, 5-trimethoxybenzylamino dithiocarboxylic acid 2- [4- [ [4- [ (3-acrylamidophenyl) amine ] -5-fluoropyrimidin-2-yl ] amine ] phenoxyethyl ester

The procedure described in preparation example 1 was followed except that the starting methylamine hydrochloride was changed to 3,4, 5-trimethoxybenzylamine with a yield of 52%.¹H NMR(400MHz,DMSO-d₆)10.47(t,J＝5.3Hz,1H),10.13(s,1H),9.38(s,1H),8.99(s,1H),8.08(d,J＝3.6Hz,1H),7.96(s,1H),7.55(d,J＝8.9Hz,2H),7.51(d,J＝7.9Hz,1H),7.43(d,J＝8.0Hz,1H),7.29(t,J＝8.1Hz,1H),6.78(d,J＝8.9Hz,2H),6.66(s,2H),6.47(dd,J＝16.9,10.1Hz,1H),6.27(dd,J＝17.0,1.6Hz,1H),5.78–5.73(m,1H),4.79(d,J＝5.3Hz,2H),4.10(t,J＝6.4Hz,2H),3.74(s,6H),3.64(s,3H),3.61(t,J＝6.4Hz,2H)。¹³C NMR(101MHz,DMSO-d₆)196.67,163.57,156.13(d,J＝2.8Hz),153.28,153.10,150.23(d,J＝10.8Hz),142.06,141.24(d,J＝19.1Hz),139.60,139.52(d,J＝6.2Hz),137.22,134.85,133.13,132.36,129.11,127.34,120.68,117.81,115.29,114.73,113.90,105.83,66.92,60.45,56.29,50.70,33.91。HRMS(ESI)m/z calcd for C₃₂H₃₄FN₆O₅S₂[M+H]⁺665.20106.Found:665.20050。

Example 17: 4-Cyanobenzylaminodithiocarboxylic acid-2- [4- [ [4- [ (3-acrylamidophenyl) amine ] -5-fluoropyrimidin-2-yl ] amine ] phenoxyethyl ester

The procedure described in preparation example 1 was followed except that the starting methylamine hydrochloride was changed to 4- (aminomethyl) benzonitrile hydrochloride in a yield of 35%.¹H NMR(400MHz,DMSO-d₆)10.65(t,J＝5.6Hz,1H),10.13(s,1H),9.38(s,1H),9.00(s,1H),8.08(d,J＝3.6Hz,1H),7.95(s,1H),7.82(d,J＝8.2Hz,2H),7.55(d,J＝8.9Hz,2H),7.51(d,J＝7.9Hz,1H),7.47(d,J＝8.2Hz,2H),7.43(d,J＝8.2Hz,1H),7.29(t,J＝8.1Hz,1H),6.79(d,J＝9.0Hz,2H),6.47(dd,J＝16.9,10.1Hz,1H),6.27(dd,J＝17.0,1.8Hz,1H),5.78–5.73(m,1H),4.94(d,J＝5.6Hz,2H),4.10(t,J＝6.3Hz,2H),3.61(t,J＝6.3Hz,2H)。¹³C NMR(101MHz,DMSO-d₆)197.72,163.57,156.13(d,J＝2.7Hz),153.07,150.23(d,J＝10.6Hz),143.59,142.05,141.24(d,J＝19.3Hz),139.61,139.52(d,J＝6.0Hz),134.87,132.82,132.36,129.11,128.78,127.34,120.68,119.26,117.82,115.29,114.76,113.89,110.38,66.82,49.63,34.06。HRMS(ESI)m/z calcd for C₃₀H₂₇FN₇O₂S₂[M+H]⁺600.16462.Found:600.16418。

Example 18: (pyridin-3-methyl) aminodithiocarboxylic acid 2- [4- [ [4- [ (3-acrylamidophenyl) amine ] -5-fluoropyrimidin-2-yl ] amine ] phenoxyethyl ester

The procedure described in preparation example 1 was followed except that the starting methylamine hydrochloride was changed to 3-aminomethylpyridine in a yield of 38%.¹HNMR(400MHz,DMSO-d₆)10.60(t,J＝5.6Hz,1H).10.13(s,1H),9.39(s,1H),8.99(s,1H),8.55(d,J＝1.8Hz,1H),8.49(dd,J＝4.8,1.6Hz,1H),8.08(d,J＝3.7Hz,1H),7.96(s,1H),7.72(dt,J＝7.8,1.9Hz,1H),7.55(d,J＝9.1Hz,2H),7.51(d,J＝8.1Hz,1H),7.43(d,J＝8.7Hz,1H),7.38(ddd,J＝7.8,4.8,0.6Hz,1H),7.29(t,J＝8.1Hz,1H),6.79(d,J＝9.1Hz,2H),6.47(dd,J＝17.0,10.1Hz,1H),6.27(dd,J＝17.0,2.0Hz,1H),5.75(dd,J＝10.1,2.0Hz,1H),4.88(d,J＝5.6Hz,2H),4.10(t,J＝6.4Hz,2H),3.61(t,J＝6.4Hz,2H)。¹³CNMR(101MHz,DMSO-d₆)197.26,163.57,156.13(d,J＝2.6Hz),153.09,150.24(d,J＝10.8Hz),149.58,148.91,142.05,141.23(d,J＝20.3Hz),139.61,139.52(d,J＝5.3Hz),136.03,134.86,133.34,132.37,129.11,127.33,124.00,120.70,117.83,115.30,114.76,113.92,66.85,47.80,33.99。HRMS(ESI)m/zcalcd for C₂₈H₂₇FN₇O₂S₂[M+H]⁺576.16462.Found:576.16394。

Example 19: (pyridin-4-methyl) aminodithiocarboxylic acid 2- [4- [ [4- [ (3-acrylamidophenyl) amine ] -5-fluoropyrimidin-2-yl ] amine ] phenoxyethyl ester

The procedure described in preparation example 1 was followed except that the starting methylamine hydrochloride was changed to 4-aminomethylpyridine in 44% yield.¹H NMR(400MHz,DMSO-d₆)10.64(t,J＝5.6Hz,1H),10.13(s,1H),9.39(s,1H),9.00(s,1H),8.53(d,J＝5.9Hz,2H),8.08(d,J＝3.7Hz,1H),7.96(s,1H),7.56(d,J＝9.0Hz,2H),7.50(s,1H),7.43(d,J＝8.1Hz,1H),7.30(d,J＝8.1Hz,1H),7.26(d,J＝5.8Hz,2H),6.79(d,J＝9.0Hz,2H),6.47(dd,J＝16.9,10.1Hz,1H),6.27(dd,J＝17.0,1.9Hz,1H),5.78–5.73(m,1H),4.89(d,J＝5.7Hz,2H),4.11(t,J＝6.4Hz,2H),3.62(t,J＝6.4Hz,2H)。¹³C NMR(101MHz,DMSO-d₆)197.93,163.57,156.13(d,J＝2.7Hz),153.08,150.23(d,J＝10.9Hz),150.08,146.72,142.06,141.26(d,J＝19.9Hz),139.61,139.52(d,J＝5.8Hz),134.88,132.37,129.12,127.35,122.78,120.68,117.82,115.29,114.78,113.90,66.84,48.93,34.08。HRMS(ESI)m/z calcd forC₂₈H₂₇FN₇O₂S₂[M+H]⁺576.16462.Found:576.16412。

Example 20: (pyridin-2-methyl) aminodithiocarboxylic acid 2- [4- [ [4- [ (3-acrylamidophenyl) amine ] -5-fluoropyrimidin-2-yl ] amine ] phenoxyethyl ester

The procedure described in preparation example 1 was followed except that the starting methylamine salt was usedThe acid salt was changed to 2-aminomethylpyridine, yield 47%.¹HNMR(400MHz,DMSO-d₆)10.62(t,J＝5.4Hz,1H),10.12(s,1H),9.38(s,1H),8.99(s,1H),8.57–8.47(m,1H),8.07(d,J＝3.6Hz,1H),7.95(s,1H),7.76(td,J＝7.7,1.6Hz,1H),7.55(d,J＝8.9Hz,2H),7.50(d,J＝7.9Hz,1H),7.41(d,J＝8.0Hz,1H),7.35–7.23(m,3H),6.78(d,J＝8.9Hz,2H),6.46(dd,J＝16.9,10.1Hz,1H),6.26(dd,J＝17.0,1.8Hz,1H),5.78–5.72(m,1H),4.93(d,J＝5.6Hz,2H),4.10(t,J＝6.3Hz,2H),3.60(t,J＝6.4Hz,2H)。¹³C NMR(101MHz,DMSO-d₆)197.41,163.57,156.95,156.13(d,J＝2.4Hz),153.09,150.23(d,J＝10.6Hz),149.55,142.05,141.24(d,J＝18.7Hz),139.60,139.52(d,J＝5.2Hz),137.21,134.85,132.36,129.12,127.36,122.88,122.02,120.69,117.83,115.30,114.76,113.92,66.88,51.92,34.00。HRMS(ESI)m/z calcd for C₂₈H₂₇FN₇O₂S₂[M+H]⁺576.16462.Found:576.16412。

Example 21: (pyrazine-2-methyl) aminodithiocarboxylic acid-2- [4- [ [4- [ (3-acrylamidophenyl) amine ] -5-fluoropyrimidin-2-yl ] amine ] phenoxyethyl ester

The procedure described in preparation example 1 was followed except that the starting methylamine hydrochloride was changed to 2-aminomethylpyrazine, yield 31%.¹H NMR(400MHz,DMSO-d₆)10.69(t,J＝5.4Hz,1H),10.13(s,1H),9.38(s,1H),8.99(s,1H),8.61(d,J＝2.1Hz,2H),8.55(d,J＝2.2Hz,1H),8.08(d,J＝3.7Hz,1H),7.96(s,1H),7.55(d,J＝9.0Hz,2H),7.50(d,J＝8.0Hz,1H),7.42(d,J＝8.1Hz,1H),7.29(t,J＝8.1Hz,1H),6.78(d,J＝9.0Hz,2H),6.47(dd,J＝17.0,10.1Hz,1H),6.26(dd,J＝17.0,1.9Hz,1H),5.75(dd,J＝9.8,2.2Hz,1H),4.97(d,J＝5.5Hz,2H),4.09(t,J＝6.4Hz,2H),3.60(t,J＝6.4Hz,2H)。¹³C NMR(101MHz,DMSO-d₆)197.74,163.56,156.11(d,J＝2.6Hz),153.06,152.76,150.23(d,J＝10.7Hz),144.64,144.29,143.88,142.04,141.23(d,J＝20.0Hz),139.59,139.51(d,J＝5.0Hz),134.85,132.36,129.11,127.35,120.67,117.82,115.29,114.75,113.90,66.79,49.81,34.07。HRMS(ESI)m/z calcd for C₂₇H₂₆FN₈O₂S₂[M+H]⁺577.15987.Found:577.15930。

Example 22: 3- [4- [ [4- [ (3-acrylamidophenyl) amine ] -5-fluoropyrimidin-2-yl ] amine ] phenoxypropyl dimethylaminothioate

Dimethylamine hydrochloride (98mg,1.2mmol) was dissolved in 15mL acetone, triethylamine (556. mu.L, 4mmol) and carbon disulfide (120. mu.L, 2mmol) were added dropwise, and the mixture was stirred at room temperature for 30 min. Then, the product of preparation example 7b (533mg,1mmol) was added and reacted at 60 ℃ for 1 hour. To the reaction system, silica gel was directly added, the solvent was dried under reduced pressure, and the mixture was separated with silica gel (P: E ═ 2:1) to obtain a white solid (189mg, yield 36%).¹H NMR(400MHz,DMSO-d₆)10.13(s,1H),9.38(s,1H),8.98(s,1H),8.07(d,J＝3.6Hz,1H),7.96(s,1H),7.55(d,J＝8.9Hz,2H),7.51(d,J＝8.0Hz,1H),7.44(d,J＝8.0Hz,1H),7.29(t,J＝8.1Hz,1H),6.77(d,J＝8.9Hz,2H),6.48(dd,J＝16.9,10.1Hz,1H),6.28(dd,J＝17.0,1.7Hz,1H),5.82–5.68(m,1H),3.96(t,J＝6.1Hz,2H),3.47(s,3H),3.37–3.30(m,5H),2.13–1.99(m,2H)。¹³C NMR(101MHz,DMSO-d₆)195.58,163.56,156.17(d,J＝2.7Hz),153.47,150.21(d,J＝10.8Hz),142.03,141.27(d,J＝21.0Hz),139.64,139.53(d,J＝7.4Hz),134.63,132.40,129.10,127.29,120.74,117.82,115.27,114.70,113.92,66.78,45.35,41.72,33.85,28.61。HRMS(ESI)m/z calcd for C₂₅H₂₈FN₆O₂S₂[M+H]⁺527.16937.Found:527.16937。

Example 23: 4-isopropylpiperazine-1-dithiocarboxylic acid-3- [4- [ [4- [ (3-acrylamidophenyl) amine ] -5-fluoropyrimidin-2-yl ] amine ] phenoxypropyl ester

The procedure described in preparation example 22 was followed except that the starting dimethylamine hydrochloride salt was changed to N-isopropylpiperazine, followed by collectingThe rate was 31%.¹H NMR(400MHz,DMSO-d₆)10.13(s,1H),9.38(s,1H),8.98(s,1H),8.07(d,J＝3.6Hz,1H),7.96(s,1H),7.56(d,J＝8.9Hz,2H),7.52(d,J＝9.3Hz,1H),7.44(d,J＝8.0Hz,1H),7.29(t,J＝8.1Hz,1H),6.78(d,J＝8.9Hz,2H),6.48(dd,J＝16.9,10.1Hz,1H),6.28(dd,J＝17.0,1.7Hz,1H),5.82–5.70(m,1H),4.22(s,2H),3.97(t,J＝6.1Hz,2H),3.88(s,2H),3.38(t,J＝7.0Hz,2H),2.68(dt,J＝12.9,6.5Hz,1H),2.50(d,J＝3.8Hz,4H),2.07(dd,J＝11.4,4.5Hz,2H),0.96(d,J＝6.5Hz,6H)。¹³C NMR(101MHz,DMSO-d₆)195.12,163.55,156.17(d,J＝2.7Hz),153.45,150.21(d,J＝10.8Hz),142.03,141.27(d,J＝18.6Hz),139.64,139.54(d,J＝8.0Hz),134.64,132.41,129.09,127.27,120.72,117.78,115.25,114.70,113.87,66.77,53.94,51.70,50.52,48.21,33.38,28.66,18.56。HRMS(ESI)m/z calcd for C₃₀H₃₇FN₇O₂S₂[M+H]⁺610.24287.Found:610.24237。

Example 24: (pyridin-3-methyl) aminodithiocarboxylic acid 3- [4- [ [4- [ (3-acrylamidophenyl) amine ] -5-fluoropyrimidin-2-yl ] amine ] phenoxypropyl ester

The procedure described in preparation example 22 was followed except that the starting dimethylamine hydrochloride salt was changed to 3-aminomethylpyridine in a yield of 32%.¹H NMR(400MHz,DMSO-d₆)10.50(t,J＝5.6Hz,1H),10.13(s,1H),9.38(s,1H),8.98(s,1H),8.54(d,J＝1.7Hz,1H),8.48(dd,J＝4.8,1.5Hz,1H),8.07(d,J＝3.7Hz,1H),7.96(s,1H),7.70(d,J＝7.8Hz,1H),7.55(d,J＝9.0Hz,2H),7.50(d,J＝8.0Hz,1H),7.43(d,J＝8.3Hz,1H),7.37(dd,J＝7.7,4.8Hz,1H),7.29(t,J＝8.1Hz,1H),6.77(d,J＝9.0Hz,2H),6.48(dd,J＝17.0,10.1Hz,1H),6.27(dd,J＝17.0,1.9Hz,1H),5.81–5.72(m,1H),4.87(d,J＝5.6Hz,2H),3.96(t,J＝6.1Hz,2H),3.38–3.34(m,2H),2.10–2.01(m,2H)。¹³C NMR(101MHz,DMSO-d₆)197.63,163.56,156.16(d,J＝2.7Hz),153.46,150.22(d,J＝10.7Hz),149.55,148.88,142.03,141.28(d,J＝18.9Hz),139.58,139.52(d,J＝7.1Hz),135.94,134.64,133.44,132.39,129.11,127.31,123.97,120.71,117.82,115.28,114.69,113.92,66.71,47.55,31.48,29.10。HRMS(ESI)m/z calcdfor C₂₉H₂₉FN₇O₂S₂[M+H]⁺590.18027.Found:590.17993。

Example 25: dimethyldithiocarbamate-4- [4- [ [4- [ (3-acrylamidophenyl) amine ] -5-fluoropyrimidin-2-yl ] amine ] phenoxybutyl ester

Dimethylamine hydrochloride (98mg,1.2mmol) was dissolved in 15mL acetone, triethylamine (556. mu.L, 4mmol) and carbon disulfide (120. mu.L, 2mmol) were added dropwise, and the mixture was stirred at room temperature for 30 min. Then, the product of preparation example 7c (547mg,1mmol) was added and reacted at 60 ℃ for 1 h. To the reaction system, silica gel was directly added, the solvent was dried under reduced pressure, and the mixture was separated with silica gel (P: E ═ 2:1) to obtain a white solid (329mg, yield 61%).¹H NMR(400MHz,DMSO-d₆)10.13(s,1H),9.38(s,1H),8.96(s,1H),8.07(d,J＝3.7Hz,1H),7.96(s,1H),7.56-7.49(m,3H),7.44(d,J＝8.3Hz,1H),7.29(t,J＝8.1Hz,1H),6.75(d,J＝9.0Hz,2H),6.48(dd,J＝17.0,10.1Hz,1H),6.28(dd,J＝17.0,1.9Hz,1H),5.79–5.72(m,1H),3.90(s,2H),3.47(s,3H),3.34(s,3H),3.29(t,J＝6.9Hz,2H),1.78(s,4H)。¹³C NMR(101MHz,DMSO-d₆)195.83,163.55,156.19(d,J＝2.7Hz),153.64,150.21(d,J＝10.8Hz),142.02,141.28(d,J＝19.6Hz),139.63,139.53(d,J＝8.1Hz),134.49,132.40,129.10,127.28,120.76,117.80,115.26,114.66,113.91,67.62,45.34,41.68,36.87,28.50,25.71。HRMS(ESI)m/z calcd forC₂₆H₃₀FN₆O₂S₂[M+H]⁺541.18502.Found:541.18506。

Example 26: (pyridin-3-methyl) aminodithiocarboxylic acid 4- [4- [ [4- [ (3-acrylamidophenyl) amine ] -5-fluoropyrimidin-2-yl ] amine ] phenoxybutyl ester

The procedure described in preparation example 25 was followed except thatDimethylaminomethane hydrochloride was changed to 3-aminomethylpyridine, and the yield was 52%.¹H NMR(400MHz,DMSO-d₆)10.46(t,J＝5.5Hz,1H),10.13(s,1H),9.38(s,1H),8.97(s,1H),8.54(d,J＝1.6Hz,1H),8.48(dd,J＝4.7,1.3Hz,1H),8.07(d,J＝3.7Hz,1H),7.96(s,1H),7.70(d,J＝7.8Hz,1H),7.54(d,J＝9.0Hz,2H),7.51(d,J＝8.6Hz,1H),7.44(d,J＝8.1Hz,1H),7.37(dd,J＝7.7,4.8Hz,1H),7.29(t,J＝8.1Hz,1H),6.75(d,J＝9.0Hz,2H),6.48(dd,J＝16.9,10.1Hz,1H),6.27(dd,J＝17.0,1.8Hz,1H),5.80–5.72(m,1H),4.87(d,J＝5.5Hz,2H),3.90(s,2H),3.29(d,J＝6.3Hz,2H),1.77(s,4H)。¹³C NMR(101MHz,DMSO-d₆)197.86,163.56,156.18(d,J＝2.7Hz),153.63,150.21(d,J＝10.8Hz),149.55,148.87,142.02,141.27(d,J＝20.0Hz),139.66,139.53(d,J＝8.0Hz),135.93,134.49,133.48,132.40,129.10,127.28,123.96,120.75,117.81,115.27,114.65,113.93,67.60,47.53,34.42,28.44,26.21。HRMS(ESI)m/z calcd forC₃₀H₃₁FN₇O₂S₂[M+H]⁺604.19592.Found:604.19580。

Evaluation of BTK kinase inhibitory Activity

In the invention, a Promega kit ADP-Glo is adopted^TMKinase inhibition assays and calculation of IC for the Compounds of the examples, Kinase Assay and BTK Kinase enzyme assays₅₀Experimental procedure 1) 1 × kinase assay buffer (40mM Tris, pH 7.5; 20mM MgCl) was diluted according to Promega kit instructions₂；0.1mg/ml BSA；2mMMnCl₂50 mu MDTT), preparing 2.5 × kinase (25 ng/. mu.L BTK, Active) and 2.5 × Substrate/ATP (Substrate, Poly (4:1Glu, Tyr) 0.5. mu.g/. mu.L; 125. mu.M Ultra-pure ATP). 2) from stock solution in the kit, adding kinase reaction system into white 384-well plate, and adding total volume 5. mu.L, wherein 1. mu.L of test compound diluent, 2. mu.L of 2.5 × kinase and 2. mu.L of 2.5 × Substrate/ATP. no kinase blank control well, adding 3. mu.L of 1 × kinase detection buffer solution and 2. mu.L of 2.5 × Substrate/ATP. no compound negative control well, adding 1. mu.L of 1 × kinase detection buffer solution, 2. mu.L of 2.5 × kinase and 2. mu.L of 2.5 × Substrate/ATP, mixing the reaction system, and adding 1. mu.L of 5. mu.5. mu.3) of Glo-L Substrate/ATP, and mixing the reaction system well at room temperature^TMReagent incubation for 30min at RTThe kinase reaction is terminated and the remaining ATP in the reaction system is consumed. 4) After 10 μ LKING ase Detection Buffer was added to each well and incubated at room temperature for 30 minutes, the chemiluminescent signal was detected using a multifunctional microplate reader. The detection reagent converts ADP in the reaction system into ATP, and a chemical light-reflecting signal generated by luciferase/luciferin reaction is used for detecting ATP, wherein the chemical light-reflecting signal is in direct proportion to the concentration of ADP in the reaction system, so that the activity of kinase is reflected. 5) Computing IC₅₀The value: in this experiment, each compound was tested for kinase inhibition at 5 concentrations, 100nM, 25nM, 6.25nM,1.56nM, 0.39nM final concentration, respectively. The kinase inhibition rate per well is (light intensity of negative well-light intensity of experimental well)/(light intensity of negative well-light intensity of blank well) × 100%. Obtaining IC by nonlinear fitting of GraphPad Prism software₅₀The value is obtained. The test results are shown in Table 1.

Evaluation of cell proliferation inhibitory Activity

The invention adopts a Promega kit

AQ_ueousNon-Radioactive cell proliferation Assay for drug inhibition of cell proliferation studies. In the experiment, Ramos cells and Raji cells, which are human Burkitt lymphoma cell lines, were cultured in 8% FBS RPMI-1640 medium, humidified at 37 ℃ and 5% CO₂The culture box is subcultured by using a T75 culture bottle, other adherent cell lines are cultured in 8% FBS DMEM culture medium, the cell culture dish is subcultured in a 37 ℃ humid culture box containing 5% CO2, the inhibition rate of the compound on the proliferation of the tumor cells is measured by using an MTS method, 1) suspended cells are taken, cells in a logarithmic growth phase are placed in a centrifuge tube, centrifuged at 1000rpm for 3 minutes, supernate is discarded, the cells are re-suspended by using 8% FBS RPMI-1640 culture medium, cell counting is carried out, and the cell concentration is adjusted to be 2 × 10⁵one/mL, and seeded in 96-well plates, 50. mu.L of cell suspension per well, drug-containing DMSO stock solution was diluted with media gradient and added to the well plates containing cells, 50. mu.L of drug dilution per well, 100. mu.L total volume of media per well, final cell concentration of 1 × 10⁴One/well, detection was performed after 48 hours of incubation. 2) Adherent cells: taking the cells in logarithmic growth phase, sucking out and culturingCulturing the culture medium in a dish, washing the cells once with PBS, adding trypsin to digest the cells, stopping digestion with 10% FBS DMEM medium, blowing the cells into a centrifuge tube, centrifuging at 1000rpm for 3 minutes, removing supernatant, resuspending the cells with 8% FBS DMEM medium, counting the cells, and adjusting the cell concentration to 5 × 10⁴cells/mL were seeded in 96-well plates and 100. mu.L of cell suspension was added to each well to a final cell concentration of 5 × 10³Per well. And (3) incubating the cells overnight, changing the culture medium after the cells are attached to the wall, sucking the culture medium of each hole, adding 100 mu L of culture medium gradient diluent of the medicine, and detecting after incubating for 48 hours. 3) Mu.l of MTS detection reagent (2mg/mLMTS,0.046mg/mL PMS) was added to each well, and after 1 hour of incubation, OD at 490nm was read using a multifunctional microplate reader. 4) Calculate IC50 value: in this experiment, each compound was tested for inhibition of cell proliferation at 5 concentration gradients and the experiment was repeated three times to obtain an average, with final concentrations of drug of 40. mu.M, 10. mu.M, 2.5. mu.M, 0.625. mu.M, and 0.156. mu.M, respectively. In the experiment, medium wells without cells were used as blank control, and wells without drug were used as negative control. The cell proliferation inhibition rate was 100% (negative control well OD value-experimental well OD value)/(negative control well OD value-blank control well OD value). Finally, the IC of the compound for inhibiting cell proliferation is obtained by nonlinear fitting by GraphPad Prism software₅₀The value is obtained. The test results are shown in Table 1.

TABLE 1 inhibitory Activity of the Compounds described in the examples on BTK kinase and on Ramos and Raji cell proliferation

As can be seen from the results in the above table, all of the compounds described in the examples had significant inhibitory effects on the proliferation of BTK kinase and Ramos and Raji cells, with the compound described in example 18 having the best effect and IC for BTK kinase₅₀IC values of 1.15nM for Ramos and Raji cells₅₀The values are 0.357 μ M and 0.706 μ M, respectively, and the inhibitory activity to kinase and the inhibitory activity to cell proliferation are superior to those of positive drugs.

Claims

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

wherein L is selected from- (CH)₂)₂-、-(CH₂)₃-、-(CH₂)₄-; and/or

2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, selected from the group consisting of:

4-isopropylpiperazine-1-dithiocarboxylic acid 4- [4- [ [4- [ (3-acrylamidophenyl) amine ] -5-fluoropyrimidin-2-yl ] amine ] phenoxybutyl ester;

(pyridin-3-methyl) aminodithiocarboxylic acid 4- [4- [ [4- [ (3-acrylamidophenyl) amine ] -5-fluoropyrimidin-2-yl ] amine ] phenoxybutyl ester.

3. A pharmaceutical composition comprising the compound of any one of claims 1 to 2 or a pharmaceutically acceptable salt thereof as an active ingredient.

4. Use of a compound according to any one of claims 1-2 or a pharmaceutically acceptable salt thereof or a pharmaceutical composition according to claim 3 in the manufacture of a medicament for modulating the catalytic activity of a tyrosine kinase, wherein the tyrosine kinase is selected from bruton's tyrosine kinase.

5. Use of a compound according to any one of claims 1-2 or a pharmaceutically acceptable salt thereof or a pharmaceutical composition according to claim 3 in the manufacture of a medicament for the treatment or prevention of cancer.

6. The use according to claim 5, wherein the cancer is chronic lymphocytic leukemia, acute lymphocytic leukemia, small lymphocytic lymphoma, mantle cell lymphoma, follicular lymphoma, diffuse large B-cell lymphoma, multiple myeloma, and Waldenstrom's macroglobulinemia.

7. A process for the preparation of a compound according to claim 1, characterized in that: