Tubulin inhibitor capable of overcoming drug resistance of paclitaxel as well as preparation method and medical application of tubulin inhibitor

Abstract

The invention discloses a tubulin inhibitor capable of overcoming drug resistance of paclitaxel as well as a preparation method and medical application of the tubulin inhibitor, and belongs to the technical field of medicinal chemistry. The tubulin inhibitor comprises a compound as shown in the specification and pharmaceutically acceptable salts, prodrugs and hydrates or solvates thereof. The tubulin inhibitor disclosed by the invention can be used for inhibiting the proliferation of paclitaxel drug-resistant tumor cells at a cellular level and is applied to medicines for treating indications related to tubulin functions, and the tubulin inhibitor disclosed by the invention hardly has obvious toxicity to normal cells and has safety. The compound can be used for treating diseases including but not limited to paclitaxel drug-resistant breast cancer, non-small cell lung cancer and the like.

Description

Technical Field

[0001] The present invention relates to a class of microtubule inhibitors that overcome paclitaxel resistance, and a preparation method and medical use thereof. As microtubule inhibitors, the compounds can inhibit the proliferation of paclitaxel-resistant tumor cells at the cellular level and can be used to treat various tumors or cancers. The invention belongs to the field of pharmaceutical chemistry technology. Background Art

[0002] Tumors seriously threaten human life and health. Paclitaxel is the first-line drug for clinical treatment of tumors, and its demand is large. However, as paclitaxel is used more and more frequently, more and more clinical tumor cells have developed paclitaxel resistance, which limits its clinical application.

[0003] Microtubules are widely present in eukaryotic cells. In the early stage of tumor cell mitosis, in order to form spindles, microtubules are in a dynamic balance of depolymerization and repolymerization. The abnormally active division and reproduction process of cancer cells is closely related to the dynamic balance of depolymerization and polymerization of microtubules / tubulin. Therefore, tubulin inhibitors bind to specific sites on tubulin, affect and interfere with the dynamic behavior of tubulin polymerization and depolymerization, and then block the formation of the M phase spindle during mitosis, causing tumor cell growth to stagnate in the G2 / M phase, thereby inhibiting the growth and proliferation of tumor cells and inducing tumor cell apoptosis. At present, there are four tubulin inhibitor binding sites on microtubules, namely, paclitaxel binding site, laulimalide binding site, vinblastine binding site and colchicine binding site. Tubulin polymerization inhibitors acting on the colchicine site can not only overcome the side effects caused by the inhibition of the other three tubulin binding sites, such as significant bone marrow suppression and neurotoxicity, but also overcome the drug resistance mechanism mediated by P-glycoprotein, and can maintain unchanged cytotoxic activity against multidrug-resistant tumor cells, and have the advantages of anti-tumor resistance. At the same time, it also has multiple functions such as being able to inhibit angiogenesis in tumor tissues to varying degrees.

[0004] Therefore, the present invention designs and synthesizes a class of microtubule protein inhibitors to overcome the problem of paclitaxel resistance, and uses it as a multifunctional microtubule protein polymerization inhibitor, which can be used to treat various paclitaxel-resistant cancers and has a good market prospect. Summary of the invention

[0005] The purpose of the present invention is to address the problems existing in the prior art and provide a microtubule protein inhibitor that can inhibit the proliferation of paclitaxel-resistant tumor cells at the cellular level and is used in drugs for treating indications related to microtubule protein function. The microtubule protein inhibitor involved in the present invention has almost no obvious toxicity to normal cells and is safe. It can be used to treat diseases including but not limited to paclitaxel-resistant breast cancer, non-small cell lung cancer, etc.

[0006] Another object of the present invention is to provide a method for preparing a class of microtubule protein inhibitors that overcome paclitaxel resistance.

[0007] Another object of the present invention is to provide a class of medical uses of tubulin inhibitors that overcome paclitaxel resistance.

[0008] In order to solve the above technical problems, the technical solution adopted by the present invention is:

[0009] The present invention discloses a class of microtubule protein inhibitors, and specifically discloses a class of microtubule protein inhibitors that overcome paclitaxel resistance. The microtubule protein inhibitors inhibit the proliferation of tumor cells by inhibiting the polymerization of microtubule protein, and are used to treat diseases including but not limited to paclitaxel-resistant breast cancer, non-small cell lung cancer, and the like.

[0010] The general structural formula of the compound of the present invention is as follows:

[0011]

[0012] Specifically, the structures of compounds T1 to T5 of the present invention are as follows:

[0013]

[0014] The present invention also includes pharmaceutically acceptable salts of compounds T1 to T5, including hydrochlorides, hydrobromides, sulfates, phosphates, methanesulfonates, acetates, lactates, succinates, fumarates, maleates, citrates, benzoates, p-toluenesulfonates or alkaline metal cation salts.

[0015] The present invention discloses a pharmaceutical composition, which comprises compounds T1-T5 or pharmaceutically acceptable salts or solvates thereof, and one or more pharmaceutically acceptable carriers, diluents or excipients.

[0016] The preparation method of the compound of the present invention is as follows:

[0017]

[0018] A method for preparing a class of microtubule protein inhibitors that overcome paclitaxel resistance comprises the following steps:

[0019] Step 1: Preparation of intermediate 4:

[0020] Take 5 g, 25 mmol of levodopa as the starting material, dissolve it in 120 mL of methanol, add 70 mmol of thionyl chloride dropwise, stir at 25 °C for 7 hours, and concentrate under reduced pressure to obtain a light yellow solid intermediate 2;

[0021] 6 g, 25 mmol of intermediate 2 was placed in 50 mL, 1,4-dioxane, 40 mL, 1 M sodium hydroxide aqueous solution was slowly added, followed by 8 g, 1.5 eq of di-tert-butyl carbonate. After reacting for 6 h, hydrochloric acid was added to adjust the pH to acidic pH = 4, and ethyl acetate was extracted three times. The ethyl acetate layers were combined and dehydrated with anhydrous sodium sulfate to obtain an oily intermediate 3, which was directly used in the next step;

[0022] The intermediate 3 was dissolved in 20 mL of acetonitrile, and 10 g, 75 mmol of potassium carbonate and 5.2 mL, 55 mmol of 1,2-dibromoethane were added. The mixture was refluxed for 8 h, and the acetonitrile was dried under reduced pressure. Water and ethyl acetate were added for separation. The aqueous layer was extracted with ethyl acetate three times. The ethyl acetate layers were combined and subjected to silica gel column chromatography. The eluent was eluted with EtAC:PE=2:5. The light yellow oily intermediate 4 was obtained after the eluent was evaporated under reduced pressure.

[0023] Step 2: Preparation of T1 to T5:

[0024] In a 25mL:25mL mixed solvent of tetrahydrofuran and methanol, add 13.5g of sodium borohydride and 16g of lithium chloride, stir for 15min, then add 2.5g, 7mmol of intermediate 4 obtained in step 1, react for 2h, filter, concentrate the filtrate under reduced pressure, add ethyl acetate to dissolve, and concentrate under reduced pressure to obtain a colorless transparent oil intermediate 5, which is directly used in the next step;

[0025] 1 g, 3.2 mmol of intermediate 5 was dissolved in glacial acetic acid, 1296 mg (5 eq) of potassium isocyanate was added, the reaction was allowed to react at room temperature overnight, 10 mL of dichloromethane was added for dilution, the pH was adjusted to 10 with potassium carbonate, the mixture was extracted with dichloromethane three times, and the mixture was concentrated under reduced pressure. The obtained oily compound 6 was dissolved in 5 M hydrochloric acid, refluxed at 60° C. for 1 hour, diluted with 20 mL of dichloromethane, the pH was adjusted to neutral with potassium carbonate, the mixture was extracted with dichloromethane, and the mixture was subjected to silica gel column chromatography (eluent: EtAc / PE=2:1) ​​to obtain a colorless powder solid compound 7.

[0026] 15 mg, 0.063 mmol of colorless solid powder compound 7 was dissolved in 20 mL of dichloromethane, 25 mg of R-benzaldehyde and 40 μL of concentrated sulfuric acid were added, and the mixture was reacted at room temperature for 6 h. The mixture was adjusted to alkalinity with sodium bicarbonate, extracted with dichloromethane, concentrated under reduced pressure, and eluted with ethyl acetate / petroleum ether = 2:3 on a silica gel column. The eluent was evaporated under reduced pressure to obtain a white solid.

[0027] Preferably, the R-based benzaldehyde includes 4-dimethylaminobenzaldehyde, 2-methyl-4-dimethylaminobenzaldehyde, 2-chloro-4-dimethylaminobenzaldehyde, 3-dimethylaminobenzaldehyde or 4-diethylaminobenzaldehyde.

[0028] Preferably, pH adjustment to alkaline refers to pH=8-9; pH adjustment to acidic refers to pH=4; reaction refers to the reflux temperature reaction of the corresponding solvent; room temperature reaction refers to the reaction at 25°C

[0029] A pharmaceutical composition is used in the preparation of a drug for treating / preventing a disease caused by abnormal microtubule protein.

[0030] Diseases caused by abnormal tubulin include breast cancer, non-small cell lung cancer, and paclitaxel-resistant breast cancer and non-small cell lung cancer.

[0031] The compounds and pharmaceutically acceptable carriers of the present invention can be administered orally or by intravenous injection in clinic. They can be used as a single drug in clinic, or in combination with other clinically used chemotherapeutic drugs and radiotherapy for the treatment of the above cancers.

[0032] The clinical dosage of the compound of the present invention is 0.01 mg-1000 mg / day, and may deviate from this range depending on the severity of the disease or the dosage form.

[0033] The present invention has the following beneficial effects:

[0034] The microtubule protein inhibitor of the present invention can inhibit the proliferation of paclitaxel-resistant tumor cells at the cellular level, and is used in drugs for treating indications related to microtubule function. The microtubule protein inhibitor involved in the present invention has almost no obvious toxicity to normal cells and is safe. It can be used to treat diseases including but not limited to paclitaxel-resistant breast cancer, non-small cell lung cancer, etc.

[0035] All the compounds of the present invention have significant proliferation inhibition activity on tumor cells breast cancer cells MCF-7, non-small cell lung cancer cells A549, paclitaxel-resistant breast cancer cells MCF-7 / TxR, paclitaxel-resistant non-small cell lung cancer cells A549 / TxR, and human normal breast epithelial cells MCF-10A, and their activities are equivalent to those of positive control drugs paclitaxel and CA-4. In particular, T1 to T5 have the same level of inhibitory activity on paclitaxel-resistant cells, and their toxicity to normal cells is much lower than that of positive control drugs CA-4 and paclitaxel, with almost no obvious toxicity.

[0036] The compounds of the present invention can well inhibit the assembly of tubulin, and their inhibitory activity is comparable to that of the positive control drug CA-4, which further indicates that the target of the compounds is the colchicine site of tubulin. DETAILED DESCRIPTION

[0037] The present invention will be further described in detail below in conjunction with specific examples. The following examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention.

[0038] Example 1

[0039]

[0040] Preparation of (6R, 10aS)-6-(4-dimethylaminophenyl)-2,3,10a,11-tetrahydro-[1,4]dioxacyclo[2,3-g]imidazo[1,5-b]isoquinoline-8,10(6H,9H)-dione (T1):

[0041]

[0042] Step 1: Preparation of intermediate 4:

[0043] Levodopa (5 g, 25 mmol) was used as the starting material, dissolved in methanol (120 mL), thionyl chloride (70 mmol) was added dropwise, and the reaction was stirred at room temperature at 25°C for 7 hours. The reaction was concentrated under reduced pressure to obtain a light yellow solid intermediate 2 with a yield of 96%.

[0044] Intermediate 2 (6 g, 25 mmol) was placed in 1,4-dioxane (50 mL), and 1 M sodium hydroxide aqueous solution (40 mL) was slowly added, followed by di-tert-butyl carbonate (8 g, 1.5 eq). After reacting for 6 h, hydrochloric acid was added to adjust to acidity (pH = 4), extracted three times with ethyl acetate, the ethyl acetate layers were combined, and the water was removed with anhydrous sodium sulfate to obtain an oily intermediate 3, which was directly used in the next step. The intermediate 3 was dissolved in acetonitrile (20 mL), potassium carbonate (10 g, 75 mmol) and 1,2-dibromoethane (5.2 mL, 55 mmol) were added, refluxed for 8 h, acetonitrile was dried under reduced pressure, water and ethyl acetate were added for separation, the water layer was extracted three times with ethyl acetate, the ethyl acetate layers were combined, and the mixture was subjected to silica gel column chromatography (ethyl acetate / petroleum ether = 2:5) to obtain a light yellow oily intermediate 4, 5.9 g, with a yield of 69%.

[0045] Step 2: Preparation of the title compound (6R, 10aS)-6-(4-dimethylaminophenyl)-2,3,10a,11-tetrahydro-[1,4]dioxacyclo[2,3-g]imidazo[1,5-b]isoquinoline-8,10(6H,9H)-dione (T1):

[0046] Sodium borohydride (13.5 g) and lithium chloride (16 g) were added to a mixed solvent of tetrahydrofuran and methanol in a ratio of 1:1 (25 mL:25 mL). After stirring for 15 min, the intermediate 4 (2.5 g, 7 mmol) obtained in step 1 was added. After reacting for 2 h, the mixture was filtered, the filtrate was concentrated under reduced pressure, and ethyl acetate was added to dissolve the mixture. The intermediate 5 was concentrated under reduced pressure to obtain a colorless transparent oily substance, which was directly used in the next step. The intermediate 5 (1 g, 3.2 mmol) was dissolved in glacial acetic acid, and 1296 mg (5 eq) of potassium isocyanate was added. The mixture was reacted at room temperature overnight, and then 10 mL of dichloromethane was added for dilution. The pH was adjusted to 10 with potassium carbonate, and the mixture was extracted with dichloromethane three times. The mixture was concentrated under reduced pressure. The obtained oily compound 6 was dissolved in 5 M hydrochloric acid, refluxed at 60° C. for 1 hour, diluted with 20 mL of dichloromethane, and the pH was adjusted to neutral with potassium carbonate. The mixture was extracted with dichloromethane and subjected to silica gel column chromatography (the eluent was ethyl acetate / petroleum ether = 2:1) to give a colorless powder solid compound 7 (460 mg, yield 63%). The intermediate 7 (15 mg, 0.063 mmol) was dissolved in dichloromethane (20 mL), and 4-dimethylaminobenzaldehyde (25 mg) was added, and concentrated sulfuric acid (40 μL) was added. The mixture was reacted at room temperature for 6 h, and then adjusted to alkaline (pH = 8) with sodium bicarbonate. The mixture was extracted with dichloromethane, concentrated under reduced pressure, and purified by silica gel column chromatography (EtAc / PE = 2:3) to obtain 10 mg of a white solid, 41%.

[0047] In this embodiment, the reaction refers to the reaction at the reflux temperature of the corresponding solvent.

[0048] After silica gel column chromatography, the eluent was evaporated under reduced pressure to obtain the product.

[0049] 1 H-NMR (500MHz, DMSO-d6) δ10.87(s,1H),7.07(d,J=8.7Hz,2H),6.80(s,1H),6.67(d,J=8.7Hz,2H),6.35(s,1H),5.80(s,1H) ,4.30(dd,J=11.4,5.1Hz,1H),4.22–4.16(m,4H),3.10(dd,J=15.6,5.1Hz,1H),2.88(s,6H),2.80(dd,J=15.5,11.4Hz,1H).

[0050] 13 C-NMR (126MHz, DMSO-d6) δ174.4,154.8,142.5,142.2,128.8,128.1,124.8,116.9,116.4,112.4,64.2,64.2,53.6,53.6,29.5.

[0051] HR-MS:[M+H]+ Calculate for C 21 H 22 N3O4,380.1605,found 380.1606.

[0052] A pharmaceutical composition comprises a class of microtubule protein inhibitors for overcoming paclitaxel resistance of this embodiment or a pharmaceutically acceptable salt or solvate thereof, and one or more pharmaceutically acceptable carriers, diluents or excipients.

[0053] A pharmaceutical composition can be administered orally or by intravenous injection with a pharmaceutically acceptable carrier.

[0054] A pharmaceutical composition is used in the preparation of a drug for treating / preventing a disease caused by abnormal microtubule protein.

[0055] Diseases caused by abnormal tubulin include breast cancer, non-small cell lung cancer, and paclitaxel-resistant breast cancer and non-small cell lung cancer.

[0056] Example 2

[0057]

[0058] Preparation of (6R, 10aS)-6-(2-methyl-4-dimethylaminophenyl)-2,3,10a,11-tetrahydro-[1,4]dioxacyclo[2,3-g]imidazo[1,5-b]isoquinoline-8,10(6H,9H)-dione (T2):

[0059] The preparation method is the same as that of Example 1. The only difference is that 4-dimethylaminobenzaldehyde is replaced by 2-methyl-4-dimethylaminobenzaldehyde. White solid, yield 43%.

[0060] 1 H-NMR (500MHz, DMSO-d6) δ10.92(s,1H),6.80(s,2H),6.55(s,2H),6.17(s,1H),5.95(s,1H),4.18(s,5H),2.86(s,11H).

[0061] 13 C-NMR(151MHz,DMSO-d6)δ174.94,155.75,149.96,142.63,136.53,129.75,129.69,129.50,12 5.28,117.15,116.09,114.21,111.09,64.55,64.53,54.78,51.04,40.56,40.51,30.15,20.35.

[0062] HR-MS:[M+H] + Calculate for C 22 H 24 N3O4,394.1780,found 394.1761.

[0063] Example 3

[0064]

[0065] Preparation of (6R, 10aS)-6-(2-chloro-4-dimethylaminophenyl)-2,3,10a,11-tetrahydro-[1,4]dioxacyclo[2,3-g]imidazo[1,5-b]isoquinoline-8,10(6H,9H)-dione (T3):

[0066] The preparation method is the same as that of Example 1. The only difference is that 4-dimethylaminobenzaldehyde is replaced by 2-chloro-4-dimethylaminobenzaldehyde. White solid, yield 47%.

[0067] 1 H-NMR (600MHz, DMSO-d6) δ10.97(s,1H),7.12(d,J=8.7Hz,1H),6.84(s,1H),6.72(s,1H),6.64(d,J=6.0Hz,1H),6.18(s,1H),6.04(s ,1H),4.47(dd,J=11.7,4.4Hz,1H),4.18(dd,J=13.9,4.2Hz,4H),3.12(dd,J=15.2,4.4Hz,1H),2.92(s,6H),2.86(d,J=11.6Hz,1H).

[0068] 13 C-NMR(151MHz,DMSO-d6)δ174.09,155.35,150.18,141.98,141.93,132.46,130.06,127.61, 127.08,124.65,116.63,114.85,111.51,111.36,63.83,63.80,55.17,51.12,39.62,29.70.

[0069] HR-MS:[M+Na] + Calculate for C 21 H 20 ClN3NaO4,436.1030,found 436.1038.Example 4

[0070]

[0071] Preparation of (6R, 10aS)-6-(3-dimethylaminophenyl)-2,3,10a,11-tetrahydro-[1,4]dioxacyclo[2,3-g]imidazo[1,5-b]isoquinoline-8,10(6H,9H)-dione (T4):

[0072] The preparation method is the same as that of Example 1. The only difference is that 4-dimethylaminobenzaldehyde is replaced by 3-dimethylaminobenzaldehyde. White solid, yield 46%.

[0073] 1 H-NMR(500MHz,DMSO-d6)δ10.90(s,1H),7.12(s,1H),6.82(s,1H),6.66(s,2H),6.48(s,1H),6.4 2(s,1H),5.81(s,1H),4.40(s,1H),4.20(s,4H),3.12(s,1H),2.89(d,J=8.1Hz,6H),2.87(s,1H).

[0074] 13 C-NMR(151MHz,DMSO-d6)δ174.33,154.99,150.47,143.62,142.35,142.03,129.06,127.56, 124.59,116.78,116.14,115.54,112.08,111.39,64.11,64.07,54.51,54.06,40.11,29.37.

[0075] HR-MS:[M+H] + Calculate for C 22 H 24 N3O4,380.1621,found 380.1623.

[0076] Example 5

[0077]

[0078] Preparation of (6R, 10aS)-6-(4-diethylaminophenyl)-2,3,10a,11-tetrahydro-[1,4]dioxacyclo[2,3-g]imidazo[1,5-b]isoquinoline-8,10(6H,9H)-dione (T5):

[0079] The preparation method is the same as that of Example 1. The only difference is that 4-dimethylaminobenzaldehyde is replaced by 4-diethylaminobenzaldehyde. White solid, yield 51%.

[0080] 1H-NMR(500MHz,DMSO-d6)δ10.86(s,1H),7.00(s,2H),6.78(s,1H),6.56(s,2H),6.36( s,1H),5.78(s,1H),4.29(s,1H),4.17(s,4H),3.08(s,1H),2.77(s,1H),1.06(s,6H).

[0081] 13 C-NMR (151MHz, DMSO-d6) δ174.44,154.82,146.83,142.43,142.16,129.41,129. 11,128.24,124.82,116.85,116.40,111.21,64.24,53.59,43.72,29.46,12.62.

[0082] HR-MS:[M+H] + Calculate for C 23 H 26 N3O4,408.1949,found 408.1933.

[0083] The following are some pharmacological experiments and results of the compounds of the present invention (T1 to T5):

[0084] 1. Anti-proliferation experiment on tumor cells

[0085] The antiproliferative activity of the compounds of the present invention on tumor cells breast cancer cells MCF-7, non-small cell lung cancer cells A549, paclitaxel-resistant breast cancer cells MCF-7 / TxR, paclitaxel-resistant non-small cell lung cancer cells A549 / TxR, and human normal breast epithelial cells MCF-10A was tested.

[0086] Methods: Cells were digested, counted, and prepared into a concentration of 5×10 4 Add 100 μL of cell suspension (3-5×10 cells / mL) to each well of a 96-well plate. 3cells). The 96-well plate was placed in a 37°C, 5% CO2 incubator for 24 hours. The drug was diluted with complete medium to the required concentration (10, 50, 250, 1250, 6250 nM), and 100 μL of the corresponding drug-containing medium was added to each well. The 96-well plate was placed in a 37°C, 5% CO2 incubator for 72 hours. 20 μL of MTT (5 mg / mL) was added to each well and continued to be cultured in the incubator for 4 hours. The culture medium was discarded, 150 μL of DMSO was added to each well to dissolve, and the mixture was gently mixed on a shaker for 10 minutes. λ = 570 nm, and the OD value of each well was read by an enzyme reader. The test results were analyzed using Graphpad Prism 6. The test results of representative compounds are shown in Table 1.

[0087] The test results showed that all compounds had significant proliferation inhibition activity against these cancer cells, and their activity was comparable to that of the positive control drugs paclitaxel and CA-4. In particular, T1-T5 had the same level of inhibitory activity against paclitaxel-resistant cells, and their toxicity to normal cells was much lower than that of the positive control drugs CA-4 and paclitaxel, with almost no obvious toxicity.

[0088] Table 1. Anti-cell proliferation activity of the compounds of the present invention

[0089]

[0090]

[0091] 2. Tubulin polymerization inhibition experiment

[0092] The compounds with better anti-cell proliferation activity among the compounds of the present invention were tested for their inhibitory activity on the polymerization of tubulin.

[0093] Methods: The selected compounds were set to 5 different concentration gradients (0.125, 0.5, 2, 4, 8 μM), and each concentration was biologically repeated 3 times. 2 mg / mL tubulin (cytoskeleton) was resuspended in PEM buffer [80 mM PIPES (pH 6.9), 0.5 mM EGTA, 2 mM MgCl2 and 15% glycerol], and then pre-incubated with the compound or solvent DMSO on ice for 5 minutes. Before detecting tubulin polymerization, PEG containing GTP was added to a final concentration of 3 mg / mL. The fluorescence was continuously detected at a wavelength of 450 nm for 60 minutes using a multifunctional fluorescence microplate reader with an excitation wavelength of 360 nm. A blank control group was set up, and the test results were analyzed by Graphpad Prism 6. The test results are shown in Table 2.

[0094] The results show that the compounds of the present invention can effectively inhibit the assembly of tubulin, and their inhibitory activity is comparable to that of the positive control drug CA-4, further indicating that the target of the compounds is the colchicine site of tubulin.

[0095] Table 2. Tubulin polymerization inhibitory activity of the compounds of the present invention

[0096]

[0097] It should be understood that in order to streamline the present disclosure and aid in understanding one or more of the various inventive aspects, in the above description of exemplary embodiments of the present invention, various features of the present invention are sometimes grouped together into a single embodiment or description thereof. However, this disclosed method should not be interpreted as reflecting an intention that the claimed invention requires more features than those expressly recited in each claim. Rather, as reflected in the claims, inventive aspects lie in less than all of the features of the previously disclosed embodiments. Therefore, the claims that follow the detailed description are hereby expressly incorporated into the detailed description, with each claim standing on its own as a separate embodiment of the present invention.

[0098] Although the present invention has been described according to a limited number of embodiments, it will be apparent to those skilled in the art, with the benefit of the above description, that other embodiments may be envisioned within the scope of the invention thus described. In addition, it should be noted that the language used in this specification is selected primarily for readability and teaching purposes, rather than for explaining or defining the subject matter of the present invention. Therefore, many modifications and variations will be apparent to those skilled in the art without departing from the scope and spirit of the appended claims. The disclosure of the present invention is illustrative, not restrictive, with respect to the scope of the present invention, which is defined by the appended claims.

[0099] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A class of microtubule protein inhibitors that overcome paclitaxel resistance, characterized in that: Including the compounds shown below and their pharmaceutically acceptable salts, prodrugs and hydrates or solvates thereof; The general structural formula of the compound is:

2. The microtubule protein inhibitor for overcoming paclitaxel resistance according to claim 1, characterized in that: Compounds include T1 to T5:

3. The microtubule protein inhibitor for overcoming paclitaxel resistance according to claim 1, characterized in that: Pharmaceutically acceptable salts include hydrochloride, hydrobromide, sulfate, phosphate, methanesulfonate, acetate, lactate, succinate, fumarate, maleate, citrate, benzoate, p-toluenesulfonate or basic metal cation salts.

4. A pharmaceutical composition, characterized in that The invention comprises a class of microtubule protein inhibitors for overcoming paclitaxel resistance as claimed in any one of claims 1 to 3 or a pharmaceutically acceptable salt or solvate thereof, and one or more pharmaceutically acceptable carriers, diluents or excipients.

5. A pharmaceutical composition according to claim 4, characterized in that: The pharmaceutically acceptable carrier is administered orally or by intravenous injection.

6. The method for preparing a class of microtubule protein inhibitors that overcome paclitaxel resistance according to any one of claims 1 to 3, characterized in that: The following steps are involved: Step 1: Preparation of intermediate 4: Take 5 g, 25 mmol of levodopa as the starting material, dissolve it in 120 mL of methanol, add 70 mmol of thionyl chloride dropwise, stir at 25 °C for 7 hours, and concentrate under reduced pressure to obtain a light yellow solid intermediate 2; 6 g, 25 mmol of intermediate 2 was placed in 50 mL of 1,4-dioxane, 40 mL of 1 M sodium hydroxide aqueous solution was slowly added, followed by 8 g, 1.5 eq of di-tert-butyl carbonate. After reacting for 6 h, hydrochloric acid was added to acidify the mixture, and the mixture was extracted with ethyl acetate three times. The ethyl acetate layers were combined and dehydrated with anhydrous sodium sulfate to obtain an oily intermediate 3, which was directly used in the next step. The intermediate 3 was dissolved in 20 mL of acetonitrile, and 10 g, 75 mmol of potassium carbonate and 5.2 mL, 55 mmol of 1,2-dibromoethane were added. The mixture was refluxed for 8 h, and the acetonitrile was dried under reduced pressure. Water and ethyl acetate were added for separation. The aqueous layer was extracted with ethyl acetate three times. The ethyl acetate layers were combined and subjected to silica gel column chromatography. The eluent was eluted with EtAC:PE=2:

5. The light yellow oily intermediate 4 was obtained after the eluent was evaporated under reduced pressure. Step 2: Preparation of T1 to T5: In a 25mL:25mL mixed solvent of tetrahydrofuran and methanol, add 13.5g of sodium borohydride and 16g of lithium chloride, stir for 15min, then add 2.5g, 7mmol of intermediate 4 obtained in step 1, react for 2h, filter, concentrate the filtrate under reduced pressure, add ethyl acetate to dissolve, and concentrate under reduced pressure to obtain a colorless transparent oil intermediate 5, which is directly used in the next step; 1 g, 3.2 mmol of intermediate 5 was dissolved in glacial acetic acid, 1296 mg, 5 eq of potassium isocyanate was added, the reaction was carried out at room temperature overnight, 10 mL of dichloromethane was added for dilution, the pH was adjusted to 10 with potassium carbonate, the mixture was extracted with dichloromethane three times, and the mixture was concentrated under reduced pressure to obtain the oily compound 6, which was dissolved in 5 M hydrochloric acid, refluxed at 60° C. for 1 hour, diluted with 20 mL of dichloromethane, the pH was adjusted to neutral with potassium carbonate, extracted with dichloromethane, and subjected to silica gel column chromatography, with the eluent being EtAc / PE=2:1, to obtain a colorless powder solid compound 7; 15 mg, 0.063 mmol of colorless solid powder compound 7 was dissolved in 20 mL of dichloromethane, 25 mg of R-benzaldehyde and 40 μL of concentrated sulfuric acid were added, and the mixture was reacted at room temperature for 6 h. The mixture was adjusted to alkalinity with sodium bicarbonate, extracted with dichloromethane, concentrated under reduced pressure, and eluted with ethyl acetate / petroleum ether = 2:3 on a silica gel column. The eluent was evaporated under reduced pressure to obtain a white solid.

7. The preparation method according to claim 6, characterized in that: The R-group benzaldehyde includes 4-dimethylaminobenzaldehyde, 2-methyl-4-dimethylaminobenzaldehyde, 2-chloro-4-dimethylaminobenzaldehyde, 3-dimethylaminobenzaldehyde or 4-diethylaminobenzaldehyde.

8. The preparation method according to claim 6, characterized in that: Adjusting pH to alkaline means pH=8-9; adjusting pH to acidic means pH=4; reaction means reaction at the reflux temperature of the corresponding solvent; room temperature reaction means reaction at 25°C.

9. Use of the pharmaceutical composition according to claim 4 in the preparation of a drug for treating / preventing a disease caused by abnormal tubulin.

10. The medical use according to claim 9, characterized in that: Diseases caused by abnormal tubulin include breast cancer, non-small cell lung cancer, and paclitaxel-resistant breast cancer and non-small cell lung cancer.

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