CN115594671A - Benzothiazole derivative and preparation method and application thereof - Google Patents

Benzothiazole derivative and preparation method and application thereof Download PDF

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CN115594671A
CN115594671A CN202110774091.4A CN202110774091A CN115594671A CN 115594671 A CN115594671 A CN 115594671A CN 202110774091 A CN202110774091 A CN 202110774091A CN 115594671 A CN115594671 A CN 115594671A
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benzothiazol
enamide
amino
ylmethoxy
pyridin
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CN115594671B (en
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刘亚婧
宫平
赵燕芳
侯云雷
秦铭泽
刘默逸
杨帆
马幸幸
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Shenyang Pharmaceutical University
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Abstract

Benzothiazole derivative shown as general formula I, geometric isomer thereof, and pharmaceutically acceptable salt, hydrate, solvate or prodrug thereof, wherein substituent R 1 、R 2 And R 3 Have the meaning given in the claims. Relates to a preparation method thereof, and also relates to a compound of the general formula I, which has irreversible inhibition on the activation of tyrosine kinase and effectively overcomes EGFR T790M drug resistance mutation, and also relates to the application of the compound, geometrical isomers thereof, and pharmaceutically acceptable salts, hydrates, solvates or prodrugs thereof in the preparation of drugs for treating EGFR mutationThe application in the preparation of medicine for treating and/or preventing lung cancer, breast cancer, gastric cancer, intestinal cancer and liver cancer.

Description

Benzothiazole derivative and preparation method and application thereof
Technical Field
The invention relates to the field of medicinal chemistry, in particular to benzothiazole derivatives, geometric isomers thereof, pharmaceutically acceptable salts, hydrates, solvates or prodrugs thereof, a preparation method thereof and application of medicinal compositions containing the benzothiazole derivatives in treating EGFR mutation diseases, especially in medicaments for treating lung cancer, breast cancer, gastric cancer, intestinal cancer and liver cancer.
Background
Malignant tumor is a common disease and frequently encountered disease seriously threatening human health, and the mortality rate of human caused by the malignant tumor accounts for the second place of all disease mortality rates, and is only second to cardiovascular and cerebrovascular diseases. Among them, lung cancer is one of the most rapidly growing malignant tumors that threaten human health and life. In many countries over the last 50 years, both lung cancer incidence and mortality have been reported to be significantly higher, with lung cancer incidence and mortality accounting for the first of all malignancies in men and lung cancer incidence and mortality accounting for the second in women. Non-small cell lung cancer (NSCLC) accounts for about 80-85% of all lung cancers, and about 75% of patients have been found to be in the middle-advanced stage with a 5-year survival rate of less than 15%.
The treatment methods of NSCLC include surgical treatment, radiation treatment, drug treatment (chemotherapy) and biological treatment, but are still mainly chemotherapy to a great extent. In recent years, with the rapid development of molecular biology, researchers have more clear understanding of the occurrence and development process of NSCLC and related targets and signaling pathways involved. At present, molecular targeted therapeutic drugs have the characteristics of strong targeting property, small toxic and side effects, remarkable curative effect and the like, and are widely applied clinically. An epidermal growth factor receptor tyrosine kinase inhibitor (EGFR-TKI) has become the most widely applied molecular targeted therapeutic drug for the clinical treatment of NSCLC.
The epidermal growth factor receptor EGFR (epidermal growth factor receptor) is an expression product of the protooncogene HER1, has ligand-induced tyrosine protein kinase activity, and is a member of the HER family. The HER family includes HER1 (ErbBl, EGFR), HER2 (ErbB 2), HER3 (ErbB 3) and HER4 (ErbB 4), all of which are located on cell membranes.
In recent years, related researches show that the occurrence of various malignant tumors is related to the abnormal expression of EGFR, and the occurrence is closely related to the complex biological behavior process of the tumors and the prognosis of patients. The possible carcinogenic mechanisms are: 1. increased expression of EGFR leads to increased downstream signaling, probably due to increased transcription at the gene level or gene amplification, etc.; 2. increased expression of the mutant EGFR receptor or ligand such that the ligand-independent tyrosine kinase domain is activated; 3. the increase of ligands over-activates the normal receptors, and many tumors can produce excess EGF and/or TGF-Q by both autocrine and paracrine, over-amplifying EGFR signaling. Therefore, overexpression of EGFR, abnormal activation of downstream signaling pathways, and abnormal functioning of target genes all contribute to the immortalization of cells to form tumors.
Clinical studies found that the sites of mutation in the EGFR gene were mainly concentrated in exons 18, 19, 20 and 21 of the intracellular TK region. Among them, deletion of exon 19 (amino acid residues 746-750, accounting for 45-50% of the total mutation) and point mutation of Leu858Arg (L858R) of exon 21 (accounting for 40-45% of the mutation) were called sensitive mutations, also called common mutations, and others were called rare mutations. It is noted that EGFR gene is subject to drug resistance mutation, and the main reason for clinical drug resistance is mutation at the T790M site. The T790M mutation is formed by replacing coded threonine by bulky methionine, so that steric hindrance is increased, the binding capacity of a receptor and TKI is reduced, the binding capacity of EGFR and ATP is improved, EGFR is in an activated state again, tumor cells continue to maintain proliferation activity, and finally the activity of an inhibitor is lost to generate drug resistance. This mutation is the main cause of drug resistance of NSCLC patients to first generation EGFR-TKIs, and the mutation frequency is about 50%.
In recent years, pharmaceutical chemists have developed a series of therapeutic drugs with different action mechanisms aiming at the target of EGFR, and the therapeutic drugs can be divided into two categories, namely monoclonal antibodies and small molecule tyrosine kinase inhibitors according to different molecular types. The monoclonal antibody can competitively inhibit the combination of EGFR and ligand, thereby inhibiting the activation and autophosphorylation of receptor, accelerating the endocytosis and degradation of cell surface receptor, reducing the formation of heterodimer, inhibiting the activation of signal conduction system, and inhibiting the proliferation of tumor cells; the action mechanism of the small molecule tyrosine kinase inhibitor is that the intracellular tyrosine kinase region is competitively combined with ATP, and the interaction between ATP and the phosphorylation site of tyrosine kinase is blocked, so that the activation of EGFR tyrosine kinase and a series of downstream signal transduction pathways are inhibited.
The EGFR monoclonal antibody brings effective treatment methods and more treatment options for lung cancer patients, and the monoclonal antibody medicines have the advantages of strong action specificity and obvious curative effect. However, the drugs have poor stability, complex preparation method and administration mode and high cost due to the adoption of a biological pharmaceutical method. In addition, in clinical application, the medicines show serious adverse reactions. These reasons limit the clinical use of such drugs.
Disclosure of Invention
Aiming at the problems in the prior art, the invention provides a benzothiazole derivative, a preparation method and application thereof, wherein the benzothiazole derivative has excellent receptor protein tyrosine kinase inhibitory activity and antitumor cell proliferative cancer activity, and the preparation method is simple and controllable.
The invention relates to benzothiazole derivatives, geometric isomers thereof, and pharmaceutically acceptable salts, hydrates, solvates or prodrugs thereof, wherein the formula I is as follows:
Figure BDA0003153718020000021
wherein,
R 1 and R 2 Identical or different, each independently represents an alkyl or cycloalkyl group, wherein the alkyl group is preferably C 1 -C 4 Alkyl, cycloalkyl is preferably C 3 -C 6 Cycloalkyl, or R 1 And R 2 Together with the nitrogen atom to which it is attached form a 5-10 membered saturated or partially saturated heterocyclyl containing 1-2 heteroatoms selected from N, O, S;
R 3 is a substituent on a benzene ring, preferably 1 to 3 substituents, and is independently selected from H, halogen, alkyl, alkoxy, trifluoromethyl, trifluoromethoxy, nitro, 5-10-membered heteroaryl C 1 -C 4 Oxy, alkyl preferably being C 1 -C 4 Alkyl, alkoxy preferably being C 1 -C 4 Alkoxy, said heteroaryl containing 1-3 heteroatoms selected from N, O, S, a 5-10 membered heteroaryl C 1 -C 4 The oxy group is preferably a pyridylmethyleneoxy group;
n is an integer selected from 1 to 3.
The present invention preferably relates to compounds of formula (I) as defined below, and pharmaceutically acceptable salts, solvates or prodrugs thereof, wherein,
R 1 、R 2 are the same or different and are each independently selected from C 1 -C 4 Alkyl radical, C 3 -C 6 Cycloalkyl, or R 1 、R 2 Together with the nitrogen atom to which it is attached form a 5-6 membered saturated or partially saturated heterocyclyl containing 1-2 heteroatoms selected from N, O;
R 3 is 1-3 substituents independently selected from H, halogen, C 1 -C 4 Alkyl radical, C 1 -C 4 Alkoxy, trifluoromethyl, trifluoromethoxy, nitro, 5-6 membered heteroarylmethylenoxy, said heteroaryl containing 1-3 heteroatoms selected from N, O, S
n is 1.
The present invention also preferably relates to compounds of formula i as defined below, geometric isomers thereof, and pharmaceutically acceptable salts, hydrates, solvates or prodrugs thereof, wherein,
R 1 、R 2 are the same or different and are each independently selected from C 1 -C 4 Alkyl radical, C 3 -C 6 Cycloalkyl, or R 1 、R 2 Together with the nitrogen atom to which it is attached form a 5-6 membered saturated or partially saturated heterocyclyl containing 1-2 heteroatoms selected from N, O;
R 3 is 1-3 substituents independently selected from H, halogen, C 1 -C 4 Alkyl radical, C 1 -C 4 Alkoxy, trifluoromethyl, trifluoromethoxy, nitro, 5-6 membered heteroarylmethylenoxy, said heteroaryl containing 1-3 heteroatoms selected from N, O, S.
The present invention particularly preferably relates to compounds of the general formula i as defined below, geometric isomers thereof, and pharmaceutically acceptable salts, hydrates, solvates or prodrugs thereof, wherein,
R 1 、R 2 are the same or different and are each independently selected from C 1 -C 4 Alkyl, or R 1 、R 2 Together with the nitrogen atom to which it is attached form a 5-6 membered saturated heterocyclyl containing 1-2 heteroatoms selected from N, O;
R 3 is 1-3 substituents independently selected from H, F, cl, methyl, methoxy, trifluoromethyl, trifluoromethoxy, nitro, pyridylmethyleneoxy;
n is 1.
Very particular preference is given according to the invention to the compounds of the following general formula I, the geometric isomers thereof, and the pharmaceutically acceptable salts, hydrates, solvates or prodrugs thereof, preferably the following compounds, without these compounds being intended to restrict the invention in any way:
(E) -4- (dimethylamino) -N- (2- ((3-fluoro-4- (pyridin-2-ylmethoxy) phenyl) amino) benzothiazol-6-yl) but-2-enamide
(E) -N- (2- ((3-chloro-4- (pyridin-2-ylmethoxy) phenyl) amino) benzothiazol-6-yl) -4- (dimethylamino) but-2-enamide
(E) -4- (dimethylamino) -N- (2- ((4- (pyridin-2-ylmethoxy) phenyl) amino) benzothiazol-6-yl) but-2-enamide
(E) -4- (dimethylamino) -N- (2- ((3-nitro-4- (pyridin-2-ylmethoxy) phenyl) amino) benzothiazol-6-yl) but-2-enamide
(E) -4- (dimethylamino) -N- (2- ((3-methyl-4- (pyridin-2-ylmethoxy) phenyl) amino) benzothiazol-6-yl) but-2-enamide
(E) -4- (dimethylamino) -N- (2- ((3-methoxy-4- (pyridin-2-ylmethoxy) phenyl) amino) benzothiazol-6-yl) but-2-enamide
(E) -N- (2- ((3-fluoro-4- (pyridin-2-ylmethoxy) phenyl) amino) benzothiazol-6-yl) -4- (piperidin-1-yl) butylbut-2-enamide
(E) -N- (2- ((3-chloro-4- (pyridin-2-ylmethoxy) phenyl) amino) benzothiazol-6-yl) -4- (piperidin-1-yl) butylbut-2-enamide
(E) -N- (2- ((3-fluoro-4- (pyridin-2-ylmethoxy) phenyl) amino) benzothiazol-6-yl) - (morpholin-4-yl) butylbut-2-enamide
(E) -N- (2- ((3-chloro-4- (pyridin-2-ylmethoxy) phenyl) amino) benzothiazol-6-yl) - (morpholin-4-yl) butylbut-2-enamide
(E) - (morpholin-4-yl) -N- (2- ((4- (pyridin-2-ylmethoxy) phenyl) amino) benzothiazol-6-yl) but-2-enamide
(E) -N- (2- ((3-methyl-4- (pyridin-2-ylmethoxy) phenyl) amino) benzothiazol-6-yl) - (morpholin-4-yl) butylbut-2-enamide
(E) -N- (2- ((3-methoxy-4- (pyridin-2-ylmethoxy) phenyl) amino) benzothiazol-6-yl) - (morpholin-4-yl) butylbut-2-enamide
(E) -N- (2- ((5-chloro-2- (pyridin-2-ylmethoxy) phenyl) amino) benzothiazol-6-yl) -4- (dimethylamino) but-2-enamide
(E) -N- (2- ((5-chloro-2- (pyridin-2-ylmethoxy) phenyl) amino) benzothiazol-6-yl) -4- (piperidin-1-yl) butylbut-2-enamide
(E) -N- (2- ((5-chloro-2- (pyridin-2-ylmethoxy) phenyl) amino) benzothiazol-6-yl) - (morpholin-4-yl) butylbut-2-enamide
(E) -N- (2- ((3-bromo-4- (pyridin-2-ylmethoxy) phenyl) amino) benzothiazol-6-yl) -4- (dimethylamino) but-2-enamide
(E) -N- (2- ((3-bromo-4- (pyridin-2-ylmethoxy) phenyl) amino) benzothiazol-6-yl) -4- (piperidin-1-yl) butylbut-2-enamide
(E) -N- (2- ((3-bromo-4- (pyridin-2-ylmethoxy) phenyl) amino) benzothiazol-6-yl) - (morpholin-4-yl) butylbut-2-enamide.
Furthermore, the benzothiazole derivatives of the above formula I may form pharmaceutically acceptable salts with acids according to conventional methods in the art. Pharmaceutically acceptable salts include the inorganic and organic acid addition salts, with the following acid addition salts being particularly preferred: hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, benzenesulfonic acid, naphthalenedisulfonic acid, acetic acid, propionic acid, lactic acid, trifluoroacetic acid, maleic acid, citric acid, fumaric acid, oxalic acid, tartaric acid, benzoic acid.
In addition, the present invention also includes prodrugs of the compounds of the present invention. Prodrugs, according to the present invention, are derivatives of compounds of formula i which may themselves have poor or even no activity, but which, upon administration, are converted under physiological conditions (e.g., by metabolism, solvolysis or otherwise) to the corresponding biologically active form.
"halogen" in the context of the present invention means fluorine or chlorine; "alkyl" refers to straight or branched chain alkyl; "cycloalkyl" refers to a substituted or unsubstituted cycloalkyl; "heteroaryl" means a monocyclic or polycyclic ring system containing one or more heteroatoms selected from N, O, S, which ring system is aromatic, such as imidazolyl, pyridyl, pyrazolyl, (1,2,3) -and (1,2,4) -triazolyl, furyl, thienyl, pyrrolyl, thiazolyl, benzothiazolyl, oxazolyl, isoxazolyl, naphthyl, quinolinyl, isoquinolinyl, benzimidazolyl, benzoxazolyl; "saturated or partially saturated heterocyclyl" means a monocyclic or polycyclic ring system containing one or more heteroatoms selected from N, O, S, such as pyrrolidinyl, morpholinyl, piperazinyl, piperidinyl, pyrazolidinyl, imidazolidinyl, phthalimidyl, and thiazolinyl.
Scheme A below describes the preparation of the general formula I of the present invention, all starting materials are prepared by the methods described in these schemes, by methods well known to those of ordinary skill in the art of organic chemistry or are commercially available. All of the final compounds of the present invention are prepared by the methods described in these schemes or by methods analogous thereto, which are well known to those of ordinary skill in the art of organic chemistry. All variable factors applied in these illustrations are as defined below or in the claims.
Figure BDA0003153718020000061
Route A Synthesis of Compounds of formula I
In scheme A, 2-amino-6-nitrobenzothiazole (1) is used as the starting material via (Boc) 2 Protecting by O, reducing, condensing with diethoxyphosphonic acid to obtain an intermediate 4, then carrying out Wittig-Horner reaction on the intermediate 4 and an intermediate 7 to obtain an intermediate 5, carrying out deprotection to obtain an intermediate 6, and finally carrying out substitution reaction on the intermediate 6 and an intermediate 8 to obtain the target compound shown in the general formula I.
Intermediate 7 was prepared as shown in scheme B by hydrolysis of feed 9 in concentrated hydrochloric acid.
Figure BDA0003153718020000062
Route B Synthesis of intermediate 7
The medicinal composition contains benzothiazole derivatives shown in the general formula I and pharmaceutically acceptable salts thereof as active ingredients, and also contains pharmaceutically acceptable carriers or excipients.
Pharmaceutically acceptable carriers for use in the pharmaceutical compositions of the present invention are of the usual type available in the pharmaceutical art and include: binders, lubricants, disintegrants, solubilizing agents, diluents, stabilizers, suspending agents, non-coloring agents, flavoring agents, etc. for oral preparations; preservatives, solubilizers, stabilizers and the like for injectable preparations; bases for topical formulations, diluents, lubricants, preservatives, and the like.
The pharmaceutical compositions of the present invention may be administered orally or parenterally (e.g., intravenously, subcutaneously, intraperitoneally, or topically), and if certain drugs are unstable in gastric conditions, they may be formulated as enteric coated tablets, which are more practical than the administration of monoclonal antibody-based drugs, and conventional formulation techniques may be used in preparing such pharmaceutical compositions.
The benzothiazole derivative or the pharmaceutically acceptable salt or the medicinal composition disclosed by the invention is applied to the preparation of medicaments for treating and/or preventing EGFR (epidermal growth factor receptor) mutant diseases, in particular to the application of medicaments for treating and/or preventing EGFR mutant tumors, and especially the application of medicaments for treating and/or preventing lung cancer, breast cancer, gastric cancer, intestinal cancer or liver cancer.
The benzothiazole derivative and the preparation method and the application thereof are subjected to the inhibition activity screening of EGFR T790M mutant kinase and T790M/L858R double mutant kinase and the anti-tumor cell proliferation activity screening of EGFR T790M/L858R double mutant non-small cell lung cancer H1975 cell strain, and the result shows that the benzothiazole derivative and the pharmaceutically acceptable salt thereof have excellent receptor protein tyrosine kinase inhibition activity and anti-tumor cell proliferation cancer activity. Therefore, the compound has good application prospect.
Detailed Description
The examples and preparations provided below further illustrate and exemplify the compounds of the present invention and their methods of preparation. It should be understood that the scope of the following examples and preparations are not intended to limit the scope of the invention in any way.
The reagents used in the experiment are all analytically pure or chemically pure; the melting points of the compounds are measured by an MP420/430 full-automatic melting point instrument manufactured by the Jinan sea energy instruments, inc., and the temperature is not corrected; measuring the mass spectrum by an Agilent 1100 type quadrupole liquid chromatography combined instrument; the hydrogen nuclear magnetic resonance spectrum was measured with a Bruker ARX-400MHz nuclear magnetic resonance analyzer.
Example 1 preparation of (E) -4- (dimethylamino) -N- (2- ((3-fluoro-4- (pyridin-2-ylmethoxy) phenyl) amino) benzothiazol-6-yl) but-2-enamide
Step 1: preparation of tert-butyl (6-nitrobenzothiazol-2-yl) carbamate (2)
10.0g (50 mmol) of 2-amino-6-nitrobenzothiazole were dissolved in 150mL of DMF at room temperature, 16.8 g (77 mmol) of Boc anhydride and 3.1g (25 mmol) of DMAP were added in this order, and the temperature was raised to 90 ℃ for 4h. After the reaction, the reaction solution was cooled to room temperature, poured into cold water to precipitate a large amount of yellow solid, filtered, washed with water to obtain a filter cake, and dried to obtain 14.8g of yellow solid with a yield of 97.8%. MS (ESI) m/z:296.1[ 2 ], [ M + H ]] + 。
Step 2: preparation of tert-butyl (6-aminobenzothiazol-2-yl) carbamate (3)
At room temperature, 10.0g (34 mmol) of intermediate 2 was added to 100mL dioxane, and the temperature was raised to 70 ℃. 1.4g (5 mmol) FeCl were added successively 3 ·6H 2 O and 0.1g (10 mmol) of activated carbon, 25.4g (508 mmol) of 80% hydrazine hydrate is added dropwise, and the temperature is raised to 80 ℃ for reaction for 2 hours after the dropwise addition. After the reaction, the reaction solution is filtered while the reaction solution is hot, the filter cake is washed by hot dioxane, the filtrate is concentrated under reduced pressure, 200mL of water is added into the residue, the mixture is stirred for 30min at room temperature, the filtration is carried out, and the filter cake is washed by water to obtain light brown solid 6.3g, wherein the yield is 70.2%. MS (ESI) m/z of 266.3[ 2 ], [ M + H ]] + 。
And step 3: preparation of tert-butyl (6- (2- (diethoxyphosphoryl) acetylamino) benzothiazol-2-yl) carbamate (4)
At room temperature, 10.0g (38 mmol) of intermediate 3 was added to 100mL of ethyl acetate, the temperature was raised to 70 ℃, a 50% mass fraction ethyl acetate solution of diethoxyphosphonic acid was added dropwise, and the reaction was carried out at 70 ℃ for 3 hours. After completion of the reaction, the reaction mixture was concentrated under reduced pressure, and 100mL of ethyl acetate, water (3X 30 mL), saturated brine (2X 30 mL) and anhydrous Na were added to the residue 2 SO 4 Drying, suction filtering and vacuum concentrating to obtain gray solid 9.1g with yield 54.5%. MS (ESI) m/z of 444.1[ 2 ], [ M ] +H] + 。
And 4, step 4: preparation of tert-butyl (E) - (6- (4- (dimethylamino) but-2-enoylamino) benzothiazol-2-yl) carbamate (5)
5.5g (34 mmol) of dimethylaminoethylacetaldehyde diethyl acetal are dissolved in 20mL of concentrated hydrochloric acid at room temperature, and reacted at 120 ℃ for 4h. After the reaction, the reaction solution was concentrated under reduced pressure to obtain brown oil for use without further purification.
5.4g (135 mmol) of sodium hydroxide was dissolved in a mixed solvent of ethanol/water (volume ratio, ethanol: water = 10). After completion of the reaction, the reaction mixture was concentrated under reduced pressure, 100mL of water was added, extraction was performed with n-butanol (3X 50 mL), and the organic layers were combined, washed with water (2X 50 mL), washed with saturated brine (2X 50 mL), and dried over anhydrous Na 2 SO 4 Drying, suction filtration and decompression concentration to obtain light yellow solid 2.9g with yield of 67.4%. MS (ESI) m/z:377.4[ 2 ] M + H] + 。
And 5: (E) Preparation of (E) -N- (2-aminobenzothiazol-6-yl) -4- (dimethylamino) but-2-enamide (6)
2.0g (5.3 mmol) of intermediate 5 are dissolved in 15mL of dichloromethane at room temperature. Trifluoroacetic acid 15mL is added dropwise under ice bath, and after the dropwise addition is finished, the reaction is carried out for 3 hours at room temperature. After the reaction is finished, concentrating the reaction solution under reduced pressure, adding 20mL of water, adjusting the pH value to 9-10 by using a saturated sodium carbonate solution, separating out the existing solid, performing suction filtration, and washing a filter cake by water to obtain 0.7g of off-white solid, wherein the yield is 47.8%. MS (ESI) m/z 275.4[ m-H ]] - 。
Step 6: preparation of 2- ((4-bromo-2-fluorophenoxy) methyl) pyridine (8)
1.7g (10 mmol) of chloromethylpyridine hydrochloride and 2.9g (20 mmol) of potassium carbonate were added to 40mL of DMF at room temperature, and the mixture was stirred at room temperature for 30min, then 2g (10 mmol) of 2-fluoro-4-bromophenol and 0.1g (0.6 mmol) of potassium iodide were added in this order, and the mixture was allowed to react at 60 ℃ for 2h. After the reaction, the reaction solution was cooled to room temperature, poured into 200mL of cold water, to precipitate a large amount of white solid, filtered, washed with water, and dried to obtain 2.9g of white solid with a yield of 99.4%. MS (ESI) m/z 282.2[ 2 ], [ M ] +H] + 。
And 7: (E) Preparation of (E) -4- (dimethylamino) -N- (2- ((3-fluoro-4- (pyridin-2-ylmethoxy) phenyl) amino) benzothiazol-6-yl) but-2-enamide
At the room temperature, the reaction kettle is used for heating,0.15g (0.5 mmol) of intermediate 6, 0.31g (1.1 mmol) of intermediate 8, 0.03g (0.16 mmol) of cuprous iodide and 0.38g (2.7 mmol) of potassium carbonate were added to 10mL of DMSO and reacted at 120 ℃ for 2h. After completion of the reaction, the reaction mixture was cooled to room temperature, 20mL of water was added, extraction was performed with methylene chloride (3X 30 mL), and the organic layers were combined, washed with water (2X 30 mL), washed with saturated brine (2X 30 mL), and anhydrous Na 2 SO 4 Drying, suction filtering and vacuum concentrating to obtain light brown solid 0.1g, yield 41.9%. m.p. 133.1-134.5 deg.C, MS (ESI) m/z 476.3[ m-H ]] - 。
1 H NMR(400MHz,DMSO-d 6 )δ10.08(s,1H),8.58(d,J＝4.4Hz,1H),7.85(td,J＝7.7,1.6 Hz,1H),7.74–7.66(m,1H),7.63(s,1H),7.61(s,1H),7.52(d,J＝7.8Hz,1H),7.36(dd,J＝7.1, 5.3Hz,1H),7.26(t,J＝8.8Hz,1H),7.17(dd,J＝11.5,1.9Hz,1H),7.10(d,J＝8.4Hz,1H),7.02 (d,J＝8.6Hz,1H),6.68(dt,J＝15.2,6.1Hz,1H),6.23(d,J＝15.3Hz,1H),5.24(s,2H),3.23(d, J＝6.0Hz,2H),2.30(s,6H).
The compounds of examples 2-19 were prepared according to the procedure for example 1, selecting the appropriate starting materials and reagents, respectively. When referring to specific reaction starting materials, it is understood that one skilled in the art can select appropriate starting materials and reagents as desired in the examples.
A benzothiazole derivative of the general formula I:
Figure BDA0003153718020000091
the structural formulae of examples 1 to 19 of the present invention are shown in the following table 1:
TABLE 1 structural formulas of examples 1 to 19
Figure BDA0003153718020000092
Figure BDA0003153718020000101
Figure BDA0003153718020000111
Example 2 preparation of (E) -N- (2- ((3-chloro-4- (pyridin-2-ylmethoxy) phenyl) amino) benzothiazol-6-yl) -4- (dimethylamino) but-2-enamide
m.p.:133.4～134.6℃，MS(ESI)m/z:492.3[M-H] - 。
1 H NMR(400MHz,DMSO-d 6 )δ10.09(s,1H),8.58(d,J＝4.5Hz,1H),7.87(t,J＝8.0Hz, 1H),7.70–7.58(m,2H),7.55(d,J＝7.7Hz,1H),7.38(s,2H),7.26(d,J＝8.6Hz,1H),7.20(d,J＝ 8.2Hz,1H),7.09(s,1H),6.77–6.57(m,1H),6.23(d,J＝15.4Hz,1H),5.27(s,2H),3.24(d,J＝ 6.0Hz,2H),2.31(s,6H).
EXAMPLE 3 preparation of (E) -4- (dimethylamino) -N- (2- ((4- (pyridin-2-ylmethoxy) phenyl) amino) benzothiazol-6-yl) but-2-enamide
m.p.:131.7～132.9℃，MS(ESI)m/z:460.3[M+H] + 。
1 H NMR(400MHz,DMSO-d 6 )δ10.04(s,1H),8.58(d,J＝4.7Hz,1H),7.83(t,J＝6.9Hz, 1H),7.59(s,2H),7.51(d,J＝7.8Hz,1H),7.35(dd,J＝6.9,5.2Hz,1H),7.27(d,J＝8.5Hz,2H), 7.06(d,J＝8.7Hz,2H),6.76–6.60(m,1H),6.21(d,J＝15.2Hz,1H),5.17(s,2H),3.18(d,J＝6.0 Hz,2H),2.27(s,6H).
Example 4 preparation of (E) -4- (dimethylamino) -N- (2- ((3-nitro-4- (pyridin-2-ylmethoxy) phenyl) amino) benzothiazol-6-yl) but-2-enamide
m.p.:138.2～140.1℃，MS(ESI)m/z:505.6[M+H] + 。
1 H NMR(600MHz,DMSO-d 6 )δ10.09(s,1H),8.57(d,J＝4.9Hz,1H),8.06(s,1H),7.87(t, J＝8.4Hz,1H),7.77(s,1H),7.62(s,1H),7.58(d,J＝9.0Hz,1H),7.53(d,J＝7.7Hz,1H),7.45(s, 1H),7.40–7.33(m,1H),7.04(d,J＝9.1Hz,1H),6.67(dt,J＝12.6,6.1Hz,1H),6.25(d,J＝15.5 Hz,1H),5.35(s,2H),3.25(s,2H),2.31(s,6H).
EXAMPLE 5 preparation of (E) -4- (dimethylamino) -N- (2- ((3-methyl-4- (pyridin-2-ylmethoxy) phenyl) amino) benzothiazol-6-yl) but-2-enamide
m.p.:137.9～138.5℃，MS(ESI)m/z:474.3[M+H] + 。
1 H NMR(600MHz,DMSO-d 6 )δ8.58(s,1H),8.07(s,1H),7.85(td,J＝7.7,1.6Hz,1H), 7.64(s,1H),7.57(s,1H),7.54(d,J＝7.8Hz,1H),7.35(dd,J＝6.9,5.2Hz,1H),7.21(s,1H),7.12 (s,2H),7.04(d,J＝8.6Hz,1H),6.67(d,J＝15.6Hz,1H),6.23(d,J＝15.4Hz,1H),5.19(s,2H), 3.51(s,2H),2.32(s,6H),2.22(s,3H).
EXAMPLE 6 preparation of (E) -4- (dimethylamino) -N- (2- ((3-methoxy-4- (pyridin-2-ylmethoxy) phenyl) amino) benzothiazol-6-yl) but-2-enamide
m.p.:135.4～136.7℃，MS(ESI)m/z:490.3[M+H] + 。
1 H NMR(400MHz,DMSO-d 6 )δ10.07(s,1H),8.57(d,J＝5.3Hz,1H),7.84(td,J＝7.7,1.7 Hz,1H),7.60(d,J＝10.5Hz,2H),7.52(d,J＝7.7Hz,1H),7.35(dd,J＝7.0,5.2Hz,1H),7.09(d, J＝8.7Hz,1H),7.06(d,J＝8.5Hz,1H),7.03(d,J＝2.0Hz,1H),6.79(d,J＝8.5Hz,1H),6.67(dt, J＝12.7,6.1Hz,1H),6.22(d,J＝15.4Hz,1H),5.15(s,2H),3.77(s,3H),3.20(d,J＝6.0Hz,2H), 2.28(s,6H).
EXAMPLE 7 preparation of (E) -N- (2- ((3-fluoro-4- (pyridin-2-ylmethoxy) phenyl) amino) benzothiazol-6-yl) -4- (piperidin-1-yl) butylbut-2-enamide
m.p.:133.1～134.5℃，MS(ESI)m/z:518.4[M+H] + 。
1 H NMR(400MHz,DMSO-d 6 )δ10.06(s,1H),8.58(d,J＝4.0Hz,1H),7.93–7.75(m,1H), 7.62(d,J＝5.8Hz,1H),7.52(d,J＝7.3Hz,2H),7.41–7.31(m,2H),7.30–7.22(m,1H),7.17(d, J＝11.5Hz,1H),7.09(d,J＝6.9Hz,1H),7.01(d,J＝7.4Hz,1H),6.68(dd,J＝15.6,5.5Hz,1H), 5.24(s,2H),3.21(s,2H),2.47(s,4H),1.54(d,J＝3.0Hz,4H),1.41(s,2H).
EXAMPLE 8 preparation of (E) -N- (2- ((3-chloro-4- (pyridin-2-ylmethoxy) phenyl) amino) benzothiazol-6-yl) -4- (piperidin-1-yl) butylbut-2-enamide
m.p.:135.7～136.9℃，MS(ESI)m/z:534.6[M+H] + 。
1 H NMR(600MHz,DMSO-d 6 )δ8.57(s,1H),8.06(s,1H),7.86(d,J＝14.7Hz,1H),7.62(s, 1H),7.54(d,J＝7.7Hz,1H),7.36(d,J＝11.3Hz,2H),7.25(d,J＝8.6Hz,1H),7.19(d,J＝8.8Hz, 1H),7.08(d,J＝8.2Hz,1H),6.68(d,J＝15.6Hz,1H),6.22(t,J＝15.2Hz,1H),5.29(s,2H),3.20 (s,2H),2.47(s,4H),1.54(s,4H),1.41(s,2H).
Example 9 preparation of (E) -N- (2- ((3-fluoro-4- (pyridin-2-ylmethoxy) phenyl) amino) benzothiazol-6-yl) - (morpholin-4-yl) butylbut-2-enamide
m.p.:140.3～141.8℃，MS(ESI)m/z:518.5[M-H] - 。
1 H NMR(600MHz,DMSO-d 6 )δ8.66–8.49(m,1H),8.06(s,1H),7.85(t,J＝8.4Hz,1H), 7.71(d,J＝8.7Hz,1H),7.53(dd,J＝16.8,8.1Hz,1H),7.48–7.43(m,1H),7.39–7.32(m,1H), 7.26(t,J＝10.5Hz,1H),7.22–7.15(m,1H),7.13(d,J＝8.5Hz,1H),7.01(d,J＝9.0Hz,1H),6.68 (d,J＝15.2Hz,1H),5.36–5.08(s,2H),3.58(d,J＝4.0Hz,4H),3.20–2.96(m,2H),2.37(s,4H).
EXAMPLE 10 preparation of (E) -N- (2- ((3-chloro-4- (pyridin-2-ylmethoxy) phenyl) amino) benzothiazol-6-yl) - (morpholin-4-yl) butylbut-2-enamide
m.p.:138.5～139.9℃，MS(ESI)m/z:536.4[M+H] + 。
1 H NMR(600MHz,DMSO-d 6 )δ10.11(s,1H),8.58(s,1H),7.86(t,J＝6.7Hz,1H),7.77(s, 1H),7.69(d,J＝7.8Hz,1H),7.54(d,J＝7.4Hz,2H),7.37(d,J＝14.7Hz,2H),7.26(d,J＝8.4 Hz,1H),7.22–7.11(m,2H),6.69(d,J＝15.2Hz,1H),5.27(s,2H),3.59(s,4H),3.12(s,2H),2.38 (s,4H).
EXAMPLE 11 preparation of (E) - (morpholin-4-yl) -N- (2- ((4- (pyridin-2-ylmethoxy) phenyl) amino) benzothiazol-6-yl) but-2-enamide
m.p.:137.2～138.8℃，MS(ESI)m/z:500.4[M-H] - 。
1 H NMR(600MHz,DMSO-d 6 )δ10.08(s,1H),8.57(d,J＝4.4Hz,1H),7.83(t,J＝7.7Hz, 1H),7.72–7.62(m,2H),7.51(s,1H),7.50(s,1H),7.34(dd,J＝11.2,6.0Hz,1H),7.26(d,J＝8.7 Hz,2H),7.14(d,J＝8.6Hz,1H),7.06(d,J＝8.8Hz,2H),6.74–6.60(m,1H),5.17(s,2H),3.71– 3.49(m,4H),3.11(d,J＝5.5Hz,2H),2.39(s,4H).
Example 12 preparation of (E) -N- (2- ((3-methyl-4- (pyridin-2-ylmethoxy) phenyl) amino) benzothiazol-6-yl) - (morpholin-4-yl) butylbut-2-enamide
m.p.:129.5～130.7℃，MS(ESI)m/z:514.4[M-H] - 。
1 H NMR(400MHz,DMSO-d 6 )δ10.09(s,1H),8.57(d,J＝4.5Hz,1H),7.85(t,J＝7.7Hz, 1H),7.71–7.62(m,2H),7.54(d,J＝14.4Hz,1H),7.40–7.30(m,2H),7.20(s,1H),7.13(t,J＝ 8.5Hz,2H),7.04(d,J＝8.6Hz,1H),6.82–6.56(m,1H),5.19(s,2H),3.87–3.47(m,4H),3.15 (d,J＝4.2Hz,2H),2.42(d,J＝2.8Hz,4H),2.21(s,3H).
Example 13 preparation of (E) -N- (2- ((3-methoxy-4- (pyridin-2-ylmethoxy) phenyl) amino) benzothiazol-6-yl) - (morpholin-4-yl) butylbut-2-enamide
m.p.:133.5～134.8℃，MS(ESI)m/z:532.4[M+H] + 。
1 H NMR(400MHz,DMSO-d 6 )δ10.07(s,1H),8.56(d,J＝4.7Hz,1H),7.83(t,J＝7.7Hz, 1H),7.73–7.62(m,2H),7.51(d,J＝7.8Hz,1H),7.42–7.29(m,1H),7.13(d,J＝8.6Hz,1H), 7.05(d,J＝8.5Hz,2H),7.02(d,J＝2.0Hz,1H),6.78(d,J＝8.4Hz,1H),6.68(d,J＝15.4Hz, 1H),5.14(s,2H),3.77(s,3H),3.67–3.47(m,4H),3.10(d,J＝5.5Hz,2H),2.38(s,4H).
Example 14 preparation of (E) -N- (2- ((5-chloro-2- (pyridin-2-ylmethoxy) phenyl) amino) benzothiazol-6-yl) -4- (dimethylamino) but-2-enamide
m.p.:151.2～152.6℃，MS(ESI)m/z:492.3[M-H] - 。
1 H NMR(400MHz,DMSO-d 6 )δ10.11(s,1H),8.58(d,J＝4.2Hz,1H),7.85(td,J＝7.7,1.5 Hz,1H),7.79(d,J＝1.2Hz,1H),7.67(dd,J＝8.4,1.1Hz,1H),7.50(d,J＝7.8Hz,1H),7.40– 7.33(m,1H),7.23(dd,J＝8.7,2.4Hz,1H),7.17(d,J＝9.3Hz,1H),7.15(d,J＝8.9Hz,1H),6.69 (dt,J＝15.1,6.1Hz,1H),6.54(d,J＝2.2Hz,1H),6.23(d,J＝15.4Hz,1H),5.28(s,2H),3.21(d, J＝5.8Hz,2H),2.28(s,6H).
EXAMPLE 15 preparation of (E) -N- (2- ((5-chloro-2- (pyridin-2-ylmethoxy) phenyl) amino) benzothiazol-6-yl) -4- (piperidin-1-yl) butylbut-2-enamide
m.p.:161.5～162.4℃，MS(ESI)m/z:534.5[M+H] + 。
1 H NMR(600MHz,DMSO-d 6 )δ10.13(s,1H),8.58(d,J＝4.3Hz,1H),8.07(s,1H),7.84(t, J＝7.7Hz,1H),7.81(s,1H),7.68(d,J＝5.0Hz,1H),7.49(d,J＝7.7Hz,1H),7.42–7.33(m,1H), 7.23(dd,J＝8.7,2.2Hz,1H),7.19(d,J＝5.7Hz,1H),7.15(d,J＝8.7Hz,1H),6.70(dd,J＝15.4, 6.1Hz,1H),6.54(s,1H),5.27(s,2H),3.26(s,2H),2.47(s,4H),1.72–1.48(m,4H),1.41(s,2H).
EXAMPLE 16 preparation of (E) -N- (2- ((5-chloro-2- (pyridin-2-ylmethoxy) phenyl) amino) benzothiazol-6-yl) - (morpholin-4-yl) butylbut-2-enamide
m.p.:162.1～163.8℃，MS(ESI)m/z:514.4[M-H] - 。
1 H NMR(400MHz,DMSO-d 6 )δ10.16(s,1H),8.58(d,J＝4.3Hz,1H),7.90–7.81(m,2H), 7.75(dd,J＝8.8,2.1Hz,1H),7.48(d,J＝7.8Hz,1H),7.40–7.33(m,1H),7.28–7.24(m,2H), 7.24(d,J＝2.3Hz,1H),7.17(d,J＝8.8Hz,1H),6.70(dt,J＝15.4,5.9Hz,1H),6.52(d,J＝2.4 Hz,1H),5.28(s,2H),3.64–3.51(m,4H),3.12(d,J＝5.2Hz,2H),2.39(s,4H).
Example 17 preparation of (E) -N- (2- ((3-bromo-4- (pyridin-2-ylmethoxy) phenyl) amino) benzothiazol-6-yl) -4- (dimethylamino) but-2-enamide
m.p.:168.2～168.9℃，MS(ESI)m/z:538.4[M-H] - 。
1 H NMR(400MHz,DMSO-d 6 )δ10.13(s,1H),8.56(d,J＝4.3Hz,1H),7.92–7.81(m,2H), 7.76(dd,J＝8.8,2.1Hz,1H),7.49(d,J＝7.8Hz,1H),7.41–7.35(m,1H),7.30–7.25(m,2H), 7.21(d,J＝2.3Hz,1H),7.19(d,J＝8.8Hz,1H),6.75(dt,J＝15.4,5.9Hz,1H),6.53(d,J＝2.4 Hz,1H),5.88(s,2H),3.61–3.52(m,4H),3.19(d,J＝5.2Hz,2H),2.38(s,4H).
EXAMPLE 18 preparation of (E) -N- (2- ((3-bromo-4- (pyridin-2-ylmethoxy) phenyl) amino) benzothiazol-6-yl) -4- (piperidin-1-yl) butylbut-2-enamide
m.p.:139.1～141.5℃，MS(ESI)m/z:578.5[M+H] + 。
1 H NMR(400MHz,DMSO-d 6 )δ10.08(s,1H),8.58(d,J＝4.0Hz,1H),7.93–7.75(m,1H), 7.65(d,J＝5.8Hz,1H),7.52(d,J＝7.3Hz,2H),7.41–7.31(m,2H),7.30–7.22(m,1H),7.17(d, J＝11.5Hz,1H),7.09(d,J＝6.9Hz,1H),7.01(d,J＝7.4Hz,1H),6.68(dd,J＝15.6,5.5Hz,1H), 5.21(s,2H),3.25(s,2H),2.49(s,4H),1.55(d,J＝3.0Hz,4H),1.41(s,2H).
EXAMPLE 19 preparation of (E) -N- (2- ((3-bromo-4- (pyridin-2-ylmethoxy) phenyl) amino) benzothiazol-6-yl) - (morpholin-4-yl) butylbut-2-enamide
m.p.:152.7～153.4℃，MS(ESI)m/z:580.4[M-H] - 。
1 H NMR(400MHz,DMSO-d 6 )δ10.16(s,1H),8.55(d,J＝4.3Hz,1H),7.90–7.81(m,2H), 7.75(dd,J＝8.8,2.1Hz,1H),7.48(d,J＝7.8Hz,1H),7.40–7.33(m,1H),7.28–7.24(m,2H), 7.24(d,J＝2.3Hz,1H),7.20(d,J＝8.8Hz,1H),6.71(dt,J＝15.4,5.9Hz,1H),6.54(d,J＝2.4 Hz,1H),5.29(s,2H),3.66–3.52(m,4H),3.12(d,J＝5.2Hz,2H),2.37(s,4H).
EXAMPLE 20 pharmacological study of the products of the invention
In vitro anti-tumor cell proliferation Activity assay
The benzothiazole derivatives of the above formula I according to the present invention were screened for anti-tumor activity in vitro.
(1) The vial was removed from the liquid nitrogen and placed in a 37 ℃ water bath for rapid thawing. And (3) sucking the liquid in the cryopreserving tube, putting the liquid in a centrifuge tube, adding 2-3mL of culture solution, centrifuging at 800rpm for 8min, collecting cells, and putting the cells into 10% fetal calf serum culture solution for culture. Changing the medium after 24h, pouring out the medium from the flask, adding 4mL of the digestion solution, and adding 5% CO at 37 ℃ 2 Beating the culture flask in the incubator for 1min, digesting the cells, pouring into a centrifuge tubeAbout 8mL of a culture medium containing 10% fetal calf serum was added to stop the digestion. Centrifuging at 800rpm for 8min, discarding supernatant, adding 4mL culture solution, blowing, mixing, and passaging according to (1/4) ratio. The cells after passage were placed in CO at 37 deg.C 2 The cells are cultured in an incubator and can be used for tests after 2 generations.
(2) Digesting the cells cultured in a culture flask to the logarithmic growth phase of the cells with pancreatin, terminating the digestion with a culture medium containing 10% serum, placing the cells in a centrifuge tube, sealing the tube, centrifuging the tube, removing the supernatant, blowing the cell suspension uniformly with the culture medium containing 10% serum, adjusting the cell suspension concentration with the culture medium, adding the cell suspension to a 96-well plate at a concentration of 100 μ L per well (A1 well is not added, the cell suspension is set as a zero-adjustment well; columns 1 and 12 are not added, and the absorbance value of the drug per se is controlled), adjusting the cell concentration to about 1 × 104/well, placing the cell suspension at 37 ℃ and 5 percent CO 2 The cells are adhered to the wall after being cultured in an incubator for 24 hours, and the cells are preferably about 30 percent of each hole under microscopic examination.
(3) The tested drugs (1-2 mg) are firstly mixed evenly by 50 mu L DMSO in a vortex mode, then culture solution containing 950 mu L10% fetal calf serum is added for dilution and even mixing, 50 mu L of the diluted drug solution is taken out and added into the first row of holes of a 24-hole plate, then 950 mu L of culture solution containing 10% serum is added into the holes with the drug solution, the mixture is evenly mixed by a pipette, 200 mu L of the culture solution is added into the second hole, 800 mu L of the culture solution containing 10% is added into the second hole for 5-fold dilution, and the like, and a total 24-hole plate is diluted into 5 drugs with different concentrations for standby. Throwing out the culture solution from the 96-well plate with embedded cells, equally dividing the 96-well plate into four regions by a cross, using the upper and lower rows of wells of the 96-well plate as blank reference wells, sequentially adding the prepared liquid medicine into the 96-well plate according to the concentration from low to high (the 96-well plate is added from right to left), adding 170 mu L of the liquid medicine into each well, adding 3 wells into each concentration, adding 170 mu L of culture solution into each well, adding the highest concentration of each adjacent medicine into the left and right rows of wells without cells, placing the wells at 37 ℃, and 5 percent CO 2 Culturing for 72h in an incubator.
(4) From CO 2 The 96-well plate was removed from the incubator, the prepared 0.5% MTT solution was added to the 96-well plate, and the plate was left at 37 ℃ with 5% CO 2 Cultured in an incubatorAnd 4h. Then, the 96-well plate was taken out, the liquid in the 96-well plate was forcibly discharged, 100. Mu.L of DMSO was added to each well, and then the plate was put on a magnetic oscillator to oscillate for 3min, to sufficiently dissolve the crystals, and the absorbance value of each well was measured by a two-wavelength method (490 nm,630 nm) on a microplate reader.
The IC of each drug was calculated from the absorbance by the Bliss method 50 The value is obtained.
IR% = (control OD-sample OD)/(control OD-blank OD). Times.100%.
The results of the in vitro antitumor cell activities of the compounds are shown in table 2.
Table 2 in vitro antitumor cell Activity of the compounds of the examples
Figure BDA0003153718020000171
In vitro protein kinase inhibitory Activity assay
(1) Compounds were tested for inhibitory activity against FLT3 and JAK2 kinase by shift rate shift assay. 1 Xkinase buffer consisting of 50mM HEPES (pH 7.5), 0.0015% Brij-35; termination buffer consisted of 100mM HEPES (pH 7.5), 0.015% Brij-35, 0.2% coating reagent #3, and 50mM EDTA.
(2) The test compound was precisely weighed and diluted to 50-fold the target concentration with 100% DMSO to prepare a stock solution. Before testing, adding 10 μ L stock solution into 90 μ L1 Xkinase buffer solution, shaking for 10min, and mixing to obtain 5 Xcompound solution; adding a certain amount of kinase into 1 Xkinase buffer solution (different concentrations of different kinases are required to be prepared) to prepare 2.5 Xkinase solution for later use; FAM-labeled polypeptide substrate and ATP were added to 1 Xkinase buffer (different concentrations of different kinase configurations were required) to make a 2.5 Xsubstrate solution for use.
(3) In a 384-well plate, 5. Mu.L of 5 XCompound solution and 10. Mu.L of 2.5 Xkinase solution were added to each well in this order, and incubated at room temperature for 10 minutes; then 10. Mu.L of 2.5 Xsubstrate solution was added and incubated at 28 ℃ for 1h; after the incubation was complete, 25. Mu.L of stop buffer was added. Reading data on a microplate reader, and calculating the inhibition rate of the kinase according to a formula:
percent inhibition% = (max-conversion)/(max-min) × 100% in the formula, max: value of no compound added wells; min: value of kinase-free wells.
The results of the in vitro inhibitory activity of the compounds against EGFR T790M mutant and against EGFR T790M/L858R double mutant protein kinase are shown in Table 3.
TABLE 3 in vitro kinase inhibitory Activity of the Compounds of the examples
Figure BDA0003153718020000181
As is clear from the above test results, the compound of formula I to be protected according to the present invention has excellent receptor protein tyrosine kinase inhibitory activity and antitumor cell proliferative cancer activity. Therefore, the compound has good application prospect.

Claims (9)

1. Benzothiazole derivatives shown in a general formula I, geometric isomers thereof, and pharmaceutically acceptable salts, hydrates, solvates or prodrugs thereof,
Figure FDA0003153718010000011
wherein,
in the formula,
R 1 and R 2 Identical or different, each independently represents alkyl or cycloalkyl, or R 1 And R 2 Together with the nitrogen atom to which it is attached form a 5-10 membered saturated or partially saturated heterocyclyl containing 1-2 heteroatoms selected from N, O, S;
R 3 is 1-3 substituents on benzene ring, and is independently selected from H, halogen, alkyl, alkoxy, trifluoromethyl, trifluoromethoxy, nitro, 5-10 membered heteroaryl C 1 -C 4 Oxy, said heteroaryl group containing 1-3 heteroatoms selected from N, O, S;
n is an integer selected from 1 to 3.
2. Benzothiazole derivatives of the general formula I as claimed in claim 1, and their geometrical isomers, pharmaceutically acceptable salts, hydrates, solvates or prodrugs, R 1 And R 2 The same or different, each independently represent C 1 -C 4 Alkyl or C 3 -C 6 Cycloalkyl radical, R 3 Independently selected from C 1 -C 4 Alkyl or C 1 -C 4 An alkoxy group.
3. Benzothiazole derivatives of the general formula I, as well as their geometric isomers, and their pharmaceutically acceptable salts, hydrates, solvates or prodrugs according to claim 1, wherein,
R 1 and R 2 Are the same or different and are each independently selected from C 1 -C 4 Alkyl radical, C 3 -C 6 Cycloalkyl, or R 1 And R 2 Together with the nitrogen atom to which it is attached form a 5-6 membered saturated or partially saturated heterocyclyl containing 1-2 heteroatoms selected from N, O;
R 3 is 1-3 substituents on benzene ring, and is independently selected from H, halogen and C 1 -C 4 Alkyl radical, C 1 -C 4 Alkoxy, trifluoromethyl, trifluoromethoxy, nitro, 5-6 membered heteroarylmethylenoxy, said heteroaryl containing 1-3 heteroatoms selected from N, O, S;
n is 1.
4. Benzothiazole derivatives of the general formula I as claimed in claim 3, and their geometrical isomers, and their pharmaceutically acceptable salts, hydrates, solvates or prodrugs,
R 1 and R 2 Are the same or different and are each independently selected from C 1 -C 4 Alkyl, or R 1 And R 2 Together with the nitrogen atom to which it is attached form a 5-6 membered saturated heterocyclyl containing 1-2 heteroatoms selected from N, O;
R 3 is 1-3 substituents independently selected from H, F, cl, methyl, methoxy, trifluoromethyl, trisFluoromethoxy, nitro, pyridylmethyleneoxy;
n is 1.
5. Benzothiazole derivatives shown in the following general formula I, and geometrical isomers, pharmaceutically acceptable salts, hydrates, solvates or prodrugs thereof, which are selected from:
(E) -4- (dimethylamino) -N- (2- ((3-fluoro-4- (pyridin-2-ylmethoxy) phenyl) amino) benzothiazol-6-yl) but-2-enamide
(E) -N- (2- ((3-chloro-4- (pyridin-2-ylmethoxy) phenyl) amino) benzothiazol-6-yl) -4- (dimethylamino) but-2-enamide
(E) -4- (dimethylamino) -N- (2- ((4- (pyridin-2-ylmethoxy) phenyl) amino) benzothiazol-6-yl) but-2-enamide
(E) -4- (dimethylamino) -N- (2- ((3-nitro-4- (pyridin-2-ylmethoxy) phenyl) amino) benzothiazol-6-yl) but-2-enamide
(E) -4- (dimethylamino) -N- (2- ((3-methyl-4- (pyridin-2-ylmethoxy) phenyl) amino) benzothiazol-6-yl) but-2-enamide
(E) -4- (dimethylamino) -N- (2- ((3-methoxy-4- (pyridin-2-ylmethoxy) phenyl) amino) benzothiazol-6-yl) but-2-enamide
(E) -N- (2- ((3-fluoro-4- (pyridin-2-ylmethoxy) phenyl) amino) benzothiazol-6-yl) -4- (piperidin-1-yl) butylbut-2-enamide
(E) -N- (2- ((3-chloro-4- (pyridin-2-ylmethoxy) phenyl) amino) benzothiazol-6-yl) -4- (piperidin-1-yl) butylbut-2-enamide
(E) -N- (2- ((3-fluoro-4- (pyridin-2-ylmethoxy) phenyl) amino) benzothiazol-6-yl) - (morpholin-4-yl) butylbut-2-enamide
(E) -N- (2- ((3-chloro-4- (pyridin-2-ylmethoxy) phenyl) amino) benzothiazol-6-yl) - (morpholin-4-yl) butylbut-2-enamide
(E) - (morpholin-4-yl) -N- (2- ((4- (pyridin-2-ylmethoxy) phenyl) amino) benzothiazol-6-yl) but-2-enamide
(E) -N- (2- ((3-methyl-4- (pyridin-2-ylmethoxy) phenyl) amino) benzothiazol-6-yl) - (morpholin-4-yl) butylbut-2-enamide
(E) -N- (2- ((3-methoxy-4- (pyridin-2-ylmethoxy) phenyl) amino) benzothiazol-6-yl) - (morpholin-4-yl) butylbut-2-enamide
(E) -N- (2- ((5-chloro-2- (pyridin-2-ylmethoxy) phenyl) amino) benzothiazol-6-yl) -4- (dimethylamino) but-2-enamide
(E) -N- (2- ((5-chloro-2- (pyridin-2-ylmethoxy) phenyl) amino) benzothiazol-6-yl) -4- (piperidin-1-yl) butylbut-2-enamide
(E) -N- (2- ((5-chloro-2- (pyridin-2-ylmethoxy) phenyl) amino) benzothiazol-6-yl) - (morpholin-4-yl) butylbut-2-enamide
(E) -N- (2- ((3-bromo-4- (pyridin-2-ylmethoxy) phenyl) amino) benzothiazol-6-yl) -4- (dimethylamino) but-2-enamide
(E) -N- (2- ((3-bromo-4- (pyridin-2-ylmethoxy) phenyl) amino) benzothiazol-6-yl) -4- (piperidin-1-yl) butylbut-2-enamide
(E) -N- (2- ((3-bromo-4- (pyridin-2-ylmethoxy) phenyl) amino) benzothiazol-6-yl) - (morpholin-4-yl) butylbut-2-enamide.
6. A pharmaceutical composition comprising as an active ingredient a benzothiazole derivative of general formula i as defined in any one of claims 1 to 5, its geometrical isomers, its pharmaceutically acceptable salts, hydrates, solvates or prodrugs together with pharmaceutically acceptable excipients.
7. Use of benzothiazole derivatives of general formula i according to any of claims 1 to 5, their geometric isomers, their pharmaceutically acceptable salts, hydrates, solvates or prodrugs, or of pharmaceutical compositions according to claim 6 for the preparation of a medicament for the treatment and/or prevention of EGFR mutation diseases.
8. Use of benzothiazole derivatives of general formula i, as well as the geometric isomers, pharmaceutically acceptable salts, hydrates, solvates or prodrugs thereof, according to any of claims 1 to 5, or of a pharmaceutical composition according to claim 6 for the preparation of a medicament for the treatment and/or prophylaxis of EGFR mutant tumors.
9. Use of a pharmaceutical composition according to any one of claims 1 to 5 or 6, or a geometric isomer thereof, or a pharmaceutically acceptable salt, hydrate, solvate or prodrug thereof, or a pharmaceutical composition according to claim 6 for the manufacture of a medicament for the treatment and/or prophylaxis of lung cancer, breast cancer, stomach cancer, intestinal cancer or liver cancer.
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