Vinylpyridine prodrug compound as well as preparation method and application thereof

Abstract

The invention discloses a vinylpyridine prodrug compound and a preparation method and application thereof.The structure of the compound is shown in the formula I. The prodrug compound specifically activates a prodrug through a palladium resin catalytic reaction, releases active ingredients under activation of palladium resin, targets microtubulin and inhibits microtubule polymerization of tumor cells; therefore, growth of tumor cells is effectively inhibited, the treatment effects of high safety, effective increase of a treatment window and generation of high efficiency and low toxicity are achieved, toxicity to normal cells can be reduced while high-efficiency treatment is achieved on tumor parts, selectivity and accuracy of treatment can be achieved, and potential wide application prospects are achieved.

Description

Technical Field

[0001] The present invention relates to a vinylpyridine prodrug compound, and also relates to a preparation method and application of the compound. Background Art

[0002] Cancer treatment has always been a focus of medical research. However, traditional chemotherapy methods are often accompanied by a wide range of side effects, especially toxicity to normal tissues. In order to improve the precision of treatment and reduce toxic side effects, targeted prodrug activation technology has received widespread attention in recent years. Bioorthogonal chemical reactions, as a technology that can activate prodrugs under specific conditions in vivo, provide new possibilities for precision treatment. Through this technology, the drug remains in a low-activity state in the body and is only activated in the specific environment of tumor cells, which can significantly reduce toxicity to normal tissues.

[0003] Microtubules are important components of the cytoskeleton and are involved in multiple biological processes such as cell division, morphology maintenance, and material transport. The dynamic stability of microtubules is particularly important for the proliferation of tumor cells. Therefore, microtubules and their related proteins have become a key target in cancer treatment. Tubulin is the main protein that makes up microtubules. It can polymerize or depolymerize to form microtubule fibers and regulate the cell cycle and cell division process. Inhibiting microtubule polymerization can prevent the normal division and proliferation of tumor cells, leading to cell cycle arrest and eventual cell death. Therefore, targeting tubulin, especially inhibiting microtubule polymerization by binding to it, has become an effective anti-cancer strategy. However, microtubule inhibitors (such as colchicine compounds) often show strong toxicity in vivo, not only inhibiting cancer cells, but also affecting normal cells, causing systemic side effects. Therefore, how to develop microtubule inhibitors that can target tumor cells and reduce toxicity to normal tissues is an important challenge in cancer treatment research. Summary of the invention

[0004] Purpose of the invention: The purpose of the present invention is to provide a vinylpyridine prodrug compound with a precise targeting effect on the colchicine binding site of tubulin, and also to provide a method for preparing the above compound and its application in the preparation of tubulin inhibitors or anti-tumor drugs.

[0005] Technical solution: The present invention discloses a vinyl pyridine prodrug compound, the structure of which is shown in Formula I:

[0006]

[0007] wherein R1, R2, R4, and R5 are each independently selected from hydrogen, halogen, hydroxyl, thiol, cyano, nitro, carboxyl, ester, acyl, amide, hydroxymethyl, aldehyde, sulfonyl, C1-C8 alkyl, C2-C8 alkenyl, C2-C8 alkynyl, C1-C4 alkoxy, C3-C6 cycloalkyl, C1-C4 alkylamino, C1-C4 imino, C6-C 10 Aryl, C3-C 10 Heteroaryl, C4-C8 heterocyclic group;

[0008] R3 is selected from hydrogen, C1-C8 alkyl, C2-C8 alkenyl, C2-C8 alkynyl, C3-C8 cycloalkyl, C6-C 10 Aryl, C3-C 10 Heteroaryl, C4-C8 heterocyclic group.

[0009] Preferably, R1, R2, R4, and R5 are each independently selected from hydrogen, halogen, hydroxyl, acyl, amide, hydroxymethyl, C1-C8 alkyl, C2-C8 alkenyl, C2-C8 alkynyl, C1-C4 alkoxy, C3-C6 cycloalkyl, C1-C4 alkylamino, C6-C 10 Aryl, C3-C 10 Heteroaryl, C4-C8 heterocyclic group;

[0010] R3 is selected from hydrogen, C1-C8 alkyl, C2-C8 alkenyl, C2-C8 alkynyl, C3-C8 cycloalkyl, C6-C 10 Aryl, C3-C 10 Heteroaryl, C4-C8 heterocyclic group.

[0011] Preferably, R1, R2, R4, and R5 are each independently selected from hydrogen, halogen, C1-C4 alkoxy, C1-C4 alkylamino, and C4-C8 heterocyclyl;

[0012] R3 is selected from hydrogen, C1-C8 alkyl, C2-C8 alkenyl, C2-C8 alkynyl, C3-C8 cycloalkyl, C6-C 10 Aryl, C3-C 10 Heteroaryl.

[0013] Preferably, R1 and R2 are selected from hydrogen, fluorine, chlorine, bromine, methoxy, ethoxy, methylamino, ethylamino, N,N-dimethylamino, tetrahydropyrrolyl;

[0014] R3 is selected from hydrogen, methyl, ethyl, allyl, propargyl, 1-butyn-3-yl, 3-methyl-1-butyn-3-yl, 1-buten-3-yl, benzyl, 4-propargyloxybenzyl, 3-propargyloxybenzyl, 2-propargyloxybenzyl, 4-allyloxybenzyl, 3-allyloxybenzyl, 2-allyloxybenzyl, 4-nitrobenzyl, 4-methoxybenzyl, 4-cyanobenzyl, 4-methylbenzyl;

[0015] R4 and R5 are selected from hydrogen, hydroxy, methoxy, ethoxy, methylamino, and N,N-dimethylamino.

[0016] Wherein, the compound also includes pharmaceutically acceptable salts.

[0017] The above compound is selected from any one of compounds 1 to 16:

[0018]

[0019]

[0020]

[0021] The preparation method of the above-mentioned vinyl pyridine prodrug compound comprises the following steps: using 2,6-dichloroisonicotinic acid as a raw material, adding a condensing agent, condensing it with N,O-dimethylhydroxylamine hydrochloride to obtain Weinreb amide, and then reducing it to obtain ketone iii, protecting the carbonyl group with 2-bromoethanol, replacing the chlorine atom with different groups to obtain intermediate vi, and deprotecting it under acidic conditions to obtain intermediate vii; intermediate vii reacts with p-toluenesulfonyl hydrazide to obtain intermediate viii; protecting differently substituted halogenated phenols with tert-butyldimethylchlorosilane to obtain intermediate x; intermediate viii and intermediate x undergo Heck reaction, and then deprotecting the group to obtain original drug xii; finally, using different halogenated hydrocarbons to perform nucleophilic substitution on the hydroxyl group to obtain the vinyl pyridine prodrug compound, and the synthesis route is as follows:

[0022]

[0023] The present invention also discloses a pharmaceutical composition, comprising the compound shown in Formula I or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0024] The above vinylpyridine prodrug compound or pharmaceutical composition can be used in the preparation of microtubule protein inhibitors.

[0025] The above vinylpyridine prodrug compound or pharmaceutical composition can also be used in the preparation of anti-tumor drugs.

[0026] Principle of the invention: The vinylpyridine prodrug compound of the present invention is a new type of bioorthogonal prodrug molecule. It targets the colchicine binding site of tubulin and uses a bioorthogonal strategy to activate the prodrug to target tubulin and inhibit the microtubule polymerization of tumor cells, thereby achieving precise targeted treatment of tumors. The molecular docking results and structure-activity relationship studies show that the hydroxyl group in the original drug molecule xii is a key group for maintaining the efficacy. By using different hydrocarbon substitutions, the interaction between the hydroxyl group and the binding pocket can be blocked to achieve a significant reduction in activity and toxicity. The prodrug molecule has extremely low activity in vivo. The palladium resin is injected into the tumor to catalyze the prodrug molecule, which can achieve specific activation of the prodrug in tumor cells, thereby releasing drugs with anti-tumor activity, thereby improving the selectivity and accuracy of the treatment. The compound of the present invention has extremely low toxicity, and the catalyst palladium resin also has good safety, and can produce an effect better than the original drug after activation. Therefore, this series of prodrug molecules can not only reduce or even avoid toxicity to normal cells, but also achieve efficient treatment at the tumor site, and has a wide range of application prospects.

[0027] Beneficial effects: Compared with the prior art, the present invention has the following significant advantages: (1) The prodrug compound of the present invention can effectively inhibit tubulin activity, has significant anti-tumor activity, has high selectivity for tumor cells, and has small toxic side effects; (2) The preparation method is simple, the reaction conditions are relatively mild, there is almost no high temperature and high pressure reaction, and the operation is convenient. There is no dangerous reaction, no highly toxic reagents, and the reaction is safe and green. The reaction yield is high, the product purification operation is simple, and it is energy-saving and efficient. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 is the structural formula of the vinylpyridine prodrug compound of the present invention;

[0029] Figure 2 The figures are the experimental results of the effects of the compounds of the present invention on the body weight of mice. DETAILED DESCRIPTION

[0030] The technical scheme of the present invention is further described below in conjunction with the examples. The test materials used in the examples can all be purchased through conventional channels.

[0031] Example 1

[0032] Compound 1: 2,6-dimethoxy-4-{1-[4-methoxy-3-(prop-2-ynyloxy)phenyl]vinyl}pyridine

[0033] Step 1: 2,6-Dichloro-N-methoxy-N-methylpyridine-4-carboxamide

[0034]

[0035] 2,6-Dichloroisonicotinic acid (4g, 20.83mmol), N,O-dimethylhydroxylamine hydrochloride (2.44g, 25.00mmol), EDCI (4.39g, 22.91mmol) and HOBt (3.09g, 22.91mmol) were dissolved in DCM, Et3N (6.83mL, 31.25mmol) was added to the reaction solution, and stirred at room temperature for 12h. After the reaction was completed, the reaction solution was filtered to remove solid impurities, and the filtrate was concentrated under reduced pressure to obtain a light yellow oily crude product. The crude product was purified by silica gel chromatography (PE:EA=4:1) to obtain the title compound as a white solid (4.35g, 88.92%). ESI-MS m / z:234.67[M+H] + .

[0036] Step 2: 1-(2,6-Dichloropyridin-4-yl)ethan-1-one

[0037]

[0038] 2,6-Dichloro-N-methoxy-N-methylpyridine-4-carboxamide (4g, 17.02mmol) was dissolved in anhydrous THF and placed in an ice bath. Under nitrogen protection, methylmagnesium bromide (3M, 5.67mL, 17.02mmol) was slowly added to the reaction flask. After 1 hour of reaction, TLC was used to monitor the reaction. After the reaction was complete, the reaction was quenched with saturated ammonium chloride solution, extracted with dichloromethane, washed with saturated brine, and the organic layer was dried over anhydrous Na2SO4. The solvent was distilled off under reduced pressure to obtain a yellow liquid (2.86g, 88.45%) without purification. ESI-MS m / z:189.81[M+H] + .

[0039] Step 3: 2,6-Dichloro-4-(2-methyl-1,3-dioxolan-2-yl)pyridine

[0040]

[0041] 1-(2,6-dichloropyridin-4-yl)ethan-1-one (2.86, 15.05mmol) and 2-bromoethanol (3.76mL, 30.10mmol) were dissolved in toluene, 1,8-diazabicyclo[5.4.0]undec-7-ene (5.62mL, 37.63mmol) was slowly added dropwise to the mixture, and stirred at 90°C for 12h. After the reaction was detected to be complete by TLC, toluene was removed under reduced pressure to obtain a crude product. The crude product was purified by silica gel chromatography (PE:EA=30:1) to obtain a white powdery solid (3.01g, 85.44%). ESI-MS m / z:234.67[M+H]+ .

[0042] Step 4: 2,6-Dimethoxy-4-(2-methyl-1,3-dioxolan-2-yl)pyridine

[0043]

[0044] In a sealed tube, 2,6-dichloro-4-(2-methyl-1,3-dioxolane-2-yl)pyridine (3.00 g, 12.82 mmol) was dissolved in 20 mL of MeOH, and solid sodium methoxide (6.92 g, 128.16 mmol) was added thereto. After stirring at 100 ° C for 12 hours, the reaction solution was cooled to room temperature and the reaction was monitored by TLC. After the reaction was completed, methanol was removed under reduced pressure. The reaction solution was extracted with ethyl acetate, the organic phase was washed with saturated brine, and dried over anhydrous Na2SO4. The solvent was removed under reduced pressure to obtain a yellow solid (2.51 g, 86.95%). ESI-MS m / z: 226.21 [M+H] + .

[0045] The crude product can be directly subjected to the next reaction.

[0046] Step 5: 1-(2,6-dimethoxypyridin-4-yl)ethan-1-one

[0047]

[0048] 2,6-dimethoxy-4-(2-methyl-1,3-dioxolan-2-yl)pyridine (2.5 g, 11.11 mmol) was dissolved in 30 mL of concentrated hydrochloric acid and reacted at 80 ° C for 1.5 h. After the reaction was completed by TCL detection, the pH of the reaction solution was adjusted to 8 using 2 mol / L NaOH solution, the reaction solution was extracted with EA, the organic phase was washed with saturated brine, and dried with anhydrous Na2SO4, and the solvent was distilled off under reduced pressure to obtain a crude product. The crude product was purified by silica gel chromatography (PE: EA = 40: 1) to obtain a white solid (1.87 g, 92.99%). ESI-MS m / z: 182.25 [M + H] + .

[0049] Step 6: N-{[(1Z)-1-(2,6-dimethoxypyridin-4-yl)ethylidene]amino}-4-methylbenzenesulfonamide

[0050]

[0051] Dissolve 1-(2,6-dimethoxypyridin-4-yl)ethan-1-one (1.80 g, 9.93 mmol) in anhydrous ethanol and slowly add 4-methylbenzenesulfonylhydrazide (2.13 g, 11.42 mmol). Stir it at 80°C for 2 hours and monitor the reaction with a TLC plate. After the reaction is complete, filter the reaction solution, take the solid and dry it to obtain the title compound as a white powder (2.84 g, 81.91%). ESI-MS m / z: 350.12 [M+H] + .

[0052] Step 7: [(5-bromo-2-methoxyphenyl)oxy]dimethyl(2-methylpropan-2-yl)silane

[0053]

[0054] Dissolve 5-bromo-2-methoxyphenol (2.30 g, 11.33 mmol) in 15 mL of DMF, add imidazole (1.90 g, 28.30 mmol) and TBSCl (3.40 g, 15.30 mmol) and stir at room temperature for 4 hours. After sufficient reaction, add water to quench the reaction. Extract the reaction solution with ethyl acetate, wash the organic phase with saturated brine, and dry it with anhydrous Na2SO4. Distill off the solvent under reduced pressure to obtain a yellow oily liquid. Purify the crude product by column chromatography using PE / EA (10:1) to obtain a colorless and transparent oily liquid (3.30 g, 91.81%). ESI-MS m / z: 237.35 [M+H] + .

[0055] Step 8: 4-[1-(3-{[dimethyl(2-methylprop-2-yl)silyl]oxy}-4-methoxyphenyl)vinyl]-2,6-dimethoxypyridine

[0056]

[0057] N-{[(1Z)-1-(2,6-dimethoxypyridin-4-yl)ethylidene]amino}-4-methylbenzenesulfonamide (1.0 g, 2.86 mmol), N-{[(1Z)-1-(2,6-dimethoxypyridin-4-yl)ethylidene]amino}-4-methylbenzenesulfonamide (908 mg, 2.86 mmol), Xphos (136 mg, 286.20 μmol), Pd(CH3CN)2Cl2 (59 mg, 228.96 μmol) and t-BuOLi (504 mg, 6.30 mmol) were added to the reaction flask. Under nitrogen protection, the raw materials were dissolved with dioxane. The reaction system was placed at 90 ° C and stirred for 5 hours. The reaction was monitored by TLC. After the reaction was complete, the reaction solution was filtered and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was purified by column chromatography using PE / EA (8:1) to obtain a light yellow transparent oily liquid (986 mg, 85.79%). ESI-MS m / z: 402.31 [M+H] + .

[0058] Step 9: 5-[1-(2,6-dimethoxypyridin-4-yl)vinyl]-2-methoxyphenol

[0059]

[0060] 4-[1-(3-{[dimethyl(2-methylprop-2-yl)silyl]oxy}-4-methoxyphenyl)vinyl]-2,6-dimethoxypyridine

[0061] (986 mg, 2.46 mmol) was dissolved in THF, and tetrabutylammonium fluoride (TBAF) (963 mg, 3.68 mmol) was slowly added to the reaction solution. After stirring for 30 minutes, the reaction was monitored by TLC plate. After the reaction was complete, the solvent was removed under reduced pressure, and the crude product was purified by column chromatography using PE / EA (6:1) to obtain a yellow solid (607 mg, 86.04%). ESI-MS m / z: 288.11 [M+H] + .

[0062] Step 10: 2,6-dimethoxy-4-{1-[4-methoxy-3-(prop-2-ynyloxy)phenyl]vinyl}pyridine

[0063] 5-[1-(2,6-dimethoxypyridin-4-yl)vinyl]-2-methoxyphenol (100 mg, 348.05 μmol), 3-bromopropyne (124.01 mg, 1.04 mmol), anhydrous potassium carbonate (144 mg, 1.04 mmol) were dissolved in acetone and heated under reflux for 12 hours. After the reaction was completed, the solvent was removed under reduced pressure, extracted with ethyl acetate, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain a crude product. The crude product was purified by column chromatography to obtain the product, a light yellow oily liquid (74 mg, 65.35%). 1 H NMR (300MHz, DMSO-d6) δ7.05(dd,J=7.5,2.0Hz,1H),6.91(d,J=2.0Hz,1H),6.84(d,J=7.5Hz,1H),6.58(s ,2H),5.73(d,J=12.4Hz,1H),5.48(d,J=12.4Hz,1H),4.81(s,2H),3.95(s,6H),3.84(s,3H),2.77(s,1H). 13 C NMR (100MHz, DMSO-d6) δ164.57,150.65,149.18,138.61,135.92,133.20,124. 48,115.02,113.80,113.71,101.89,80.82,78.21,59.21,56.50,55.14.ESI-MS m / z:326.41[M+H] + .

[0064] Example 2

[0065] Compound 2: 2,6-dimethoxy-4-{1-[4-methoxy-3-(prop-2-enyloxy)phenyl]vinyl}pyridine

[0066] Using 5-[1-(2,6-dimethoxypyridin-4-yl)vinyl]-2-methoxyphenol (100 mg, 348.05 μmol), 3-bromopropylene and anhydrous potassium carbonate as raw materials, the reaction was carried out according to step 10 of Example 1 to obtain a light yellow oily liquid (79 mg, 69.33%). 1H NMR (300MHz, DMSO-d6) δ7.06(dd,J=7.5,2.0Hz,1H),6.95(d,J=2.0Hz,1H),6.83(d,J=7.5Hz,1H),6.58(s,2H),5.99(s,1H),5.73 (d,J=12.4Hz,1H),5.48(d,J=12.4Hz,1H),5.38(d,J=12.4Hz,1H),5.13(d,J=12.4Hz,1H),4.54(s,2H),3.95(s,6H),3.84(s,3H). 13 C NMR(100MHz,DMSO-d6)δ164.57,151.64,150.03,138.60,135.68,133.20,133.1 3,124.52,118.88,115.02,113.19,112.42,101.89,74.88,56.50,55.14.ESI-MS m / z:328.26[M+H] + .

[0067] Example 3

[0068] Compound 3: 4-{1-[3-(benzyloxy)-4-methoxyphenyl]vinyl}-2,6-dimethoxypyridine

[0069] Using 5-[1-(2,6-dimethoxypyridin-4-yl)vinyl]-2-methoxyphenol (100 mg, 348.05 μmol), benzyl bromide and anhydrous potassium carbonate as raw materials, the reaction was carried out according to step 10 of Example 1 to obtain a light yellow oily liquid (98 mg, 74.60%). 1 HNMR (300MHz, DMSO-d6) δ7.40–7.24(m,5H),7.06(dd,J=7.5,2.0Hz,1H),6.93(d,J=2.0Hz,1H),6.83(d,J=7.5Hz, 1H), 6.58 (s, 2H), 5.73 (d, J = 12.4Hz, 1H), 5.48 (d, J = 12.5Hz, 1H), 5.10 (t, J = 1.0Hz, 2H), 3.95 (s, 6H), 3.84 (s, 3H). 13C NMR(100MHz,DMSO-d6)δ164.57,151.45,150.23,140.03,138.60,135.75,133.20,132.4 7,132.28,132.20,124.52,115.02,113.19,111.91,101.89,81.70,56.50,55.12ESI-MS m / z:378.24[M+H] + .

[0070] Example 4

[0071] Compound 4: 4-{1-[3-(but-3-yn-2-yloxy)-4-methoxyphenyl]vinyl}-2,6-dimethoxypyridine

[0072] 5-[1-(2,6-Dimethoxypyridin-4-yl)vinyl]-2-methoxyphenol (150 mg, 522.08 μmol), 3-butyn-2-ol (43 mg, 622.49 μmol), and triphenylphosphine (274 mg, 1.04 mmol) were dissolved in anhydrous dichloromethane, cooled to 0°C, protected by nitrogen, and diisopropyl azodicarboxylate (211 mg, 1.04 mmol) was added dropwise. The reaction was allowed to react at room temperature for 3 hours, and the reaction was quenched with water, extracted with dichloromethane, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain a crude product, which was purified by column chromatography to obtain the target product as a colorless oily liquid (101 mg, 57.00%). 1 H NMR (300MHz, DMSO-d6) δ7.05 (dd, J=7.5, 2.0Hz, 1H), 6.87–6.81 (m, 2H), 6.58 (s, 2H), 5.73 (d, J=12. 4Hz,1H),5.48(d,J=12.4Hz,1H),4.87(s,1H),3.95(s,6H),3.84(s,3H),2.60(s,1H),1.49(s,3H). 13 C NMR(100MHz,DMSO-d6)δ164.57,150.52,149.62,138.61,135.64,133.20,124.45, 115.02,113.36,112.32,101.89,80.65,74.96,74.68,56.50,55.14,21.70.ESI-MS m / z:340.25[M+H] + .

[0073] Example 5

[0074] Compound 5: 2,6-dimethoxy-4-(1-{4-methoxy-3-[(2-methylbut-3-yn-2-yl)oxy]phenyl}vinyl)pyridine

[0075] Using 5-[1-(2,6-dimethoxypyridin-4-yl)vinyl]-2-methoxyphenol (100 mg, 348.05 μmol), 3-chloro-3-methylbut-1-yne and anhydrous potassium carbonate as raw materials, the reaction was carried out according to step 10 of Example 1 to obtain a light yellow oily liquid (59 mg, 47.96%). 1 H NMR (300MHz, DMSO-d6) δ7.05(dd,J=7.5,2.0Hz,1H),6.98(d,J=2.0Hz,1H),6.85(d,J=7.5Hz,1H),6.58(s ,2H),5.73(d,J=12.4Hz,1H),5.48(d,J=12.4Hz,1H),3.95(s,6H),3.84(s,3H),2.52(s,1H),1.47(s,6H). 13 C NMR(100MHz,DMSO-d6)δ164.57,148.57,148.46,138.63,135.30,133.20,124.90, 115.02,113.18,112.00,101.89,86.18,80.10,74.43,56.50,55.14,30.19.ESI-MS m / z:354.26[M+H] + .

[0076] Example 6

[0077] Compound 6: 4-{1-[3-(but-3-en-2-yloxy)-4-methoxyphenyl]vinyl}-2,6-dimethoxypyridine

[0078] Using 5-[1-(2,6-dimethoxypyridin-4-yl)vinyl]-2-methoxyphenol (100 mg, 348.05 μmol), 3-chloro-3-methylbut-1-yne and anhydrous potassium carbonate as raw materials, the reaction was carried out according to step 10 of Example 1 to obtain a light yellow oily liquid (81 mg, 68.17%). 1H NMR (300MHz, DMSO-d6) δ7.05(dd,J=7.5,2.0Hz,1H),6.88(d,J=2.0Hz,1H),6.84(d,J=7.5Hz,1H),6.58(s,2H),5.73(d,J= 12.4Hz,1H),5.58(s,1H),5.51-5.41(m,2H),5.19(d,J=12.4Hz,1H),5.07(s,1H),3.95(s,6H),3.84(s,3H),1.38(s,3H). 13 C NMR(100MHz,DMSO-d6)δ164.57,150.38,150.00,138.60,137.70,135.43,133.20,1 24.52,115.75,115.02,113.06,112.99,101.89,78.06,56.50,55.14,21.87.ESI-MS m / z:342.55[M+H] + .

[0079] Example 7

[0080] Compound 7: 2,6-dimethoxy-4-{1-[4-methoxy-3-({[2-(prop-2-ynyloxy)phenyl]methyl}oxy)phenyl]vinyl}pyridine

[0081] Step 1: [2-(Prop-2-ynyloxy)phenyl]methanol

[0082]

[0083] 2-Hydroxymethylphenol (300 mg, 2.42 mmol), 3-bromopropyne (862 mg, 7.25 mmol), and anhydrous potassium carbonate (1.34 g, 9.67 mmol) were dissolved in acetone and heated under reflux for 12 hours. After the reaction was completed, the solvent was removed under reduced pressure, extracted with ethyl acetate, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain a crude product. The crude product was purified by column chromatography to obtain the title compound (315 mg, 80.37%). ESI-MS m / z: 163.31 [M+H] + .

[0084] Step 2: 1-(Bromomethyl)-2-(prop-2-ynyloxy)benzene

[0085]

[0086] [2-(prop-2-ynyloxy)phenyl]methanol (315mg, 1.94mmol) was dissolved in dichloromethane and placed in an ice bath. Phosphorus tribromide (365μL, 3.88mmol) was added dropwise. After reacting at 0°C for 1 hour, TLC was used for monitoring. After the reaction was complete, the reaction was quenched with a saturated sodium bicarbonate solution, extracted with dichloromethane, washed with saturated brine, and dried over anhydrous Na2SO4. The solvent was distilled off under reduced pressure to obtain a yellow oily crude product. The crude product was purified by column chromatography to obtain the product (281mg, 64.28%). ESI-MS m / z: 224.71[M+H] + .

[0087] Step 3: 2,6-dimethoxy-4-{1-[4-methoxy-3-({[2-(prop-2-ynyloxy)phenyl]methyl}oxy)phenyl]vinyl}pyridine

[0088] Using 5-[1-(2,6-dimethoxypyridin-4-yl)vinyl]-2-methoxyphenol, 1-(bromomethyl)-2-(prop-2-ynyloxy)benzene and anhydrous potassium carbonate as raw materials, follow the steps of Step 10 of Example 1 to obtain a white solid (98 mg, 65.26%). 1 H NMR(300MHz,DMSO-d6)δ7.35(ddt,J=7.5,2.1,1.0Hz,1H),7.29–7.23(m,1H),7.09–7.01(m,2H),6.92–6.81(m,3H),6.58(s,2 H),5.73(d,J=12.4Hz,1H),5.48(d,J=12.5Hz,1H),5.10(d,J=0.9Hz,2H),4.76(s,2H),3.95(s,6H),3.84(s,3H),2.68(s,1H). 13 CNMR(100MHz,DMSO-d6)δ164.57,162.05,150.58,150.47,138.60,135.75,133.22,133.20,131.44,128.15, 125.57,124.52,116.44,115.02,113.19,112.50,101.89,80.53,78.61,78.28,60.84,56.50,55.14.ESI-MS m / z:432.66[M+H] + .

[0089] Example 8

[0090] Compound 8: 2,6-dimethoxy-4-{1-[4-methoxy-3-({[3-(prop-2-ynyloxy)phenyl]methyl}oxy)phenyl]vinyl}pyridine

[0091] Using 3-hydroxymethylphenol and 3-bromopropyne as raw materials and following the procedure of Example 7, a white solid (84 mg, 55.93%) was obtained. 1 H NMR(300MHz,DMSO-d6)δ7.27(dtd,J=5.9,2.0,1.0Hz,1H),7.22(t,J=7.4Hz,1H ),7.06(dd,J=7.5,2.0Hz,1H),6.93(d,J=2.0Hz,1H),6.88–6.81(m,2H),6.70( tt,J=2.0,1.0Hz,1H),6.58(s,2H),5.73(d,J=12.4Hz,1H),5.48(d,J=12.5Hz, 1H), 5.02 (t, J = 1.0Hz, 2H), 4.73 (s, 2H), 3.95 (s, 6H), 3.84 (s, 3H), 2.68 (s, 1H). 13 C NMR(100MHz,DMSO-d6)δ164.57,158.06,150.95,150.42,138.60,136.67,135.75,133.20,131.10,125.98, 124.52,118.22,115.70,115.02,113.19,111.91,101.89,80.74,80.07,78.28,60.45,56.50,55.14.ESI-MS m / z:432.35[M+H] + .

[0092] Example 9

[0093] Compound 9: 2,6-dimethoxy-4-{1-[4-methoxy-3-({[4-(prop-2-ynyloxy)phenyl]methyl}oxy)phenyl]vinyl}pyridine

[0094] Using 4-hydroxymethylphenol and 3-bromopropyne as raw materials and following the procedure of Example 7, a white solid (109 mg, 72.58%) was obtained. 1H NMR (300MHz, DMSO-d6) δ7.29–7.22(m,2H),7.06(dd,J=7.5,2.0Hz,1H),6.93(d,J=2.0Hz,1H),6.86–6.80(m,3H),6.58(s,2H ),5.73(d,J=12.4Hz,1H),5.48(d,J=12.5Hz,1H),5.11(t,J=1.0Hz,2H),4.68(s,2H),3.95(s,6H),3.84(s,3H),2.68(s,1H). 13 CNMR(100MHz,DMSO-d6)δ164.57,161.59,151.45,150.27,138.60,135.75,134.34,133.20,133.08, 124.52,117.02,115.02,113.19,111.91,101.89,81.20,80.74,78.28,59.69,56.50,55.14.ESI-MS m / z:432.61[M+H] + .

[0095] Example 10

[0096] Compound 10: 2,6-dimethoxy-4-{1-[4-methoxy-3-({[2-(prop-2-enyloxy)phenyl]methyl}oxy)phenyl]vinyl}pyridine

[0097] Using 2-hydroxymethylphenol and 3-bromopropylene as raw materials and following the procedure of Example 7, a white solid (84 mg, 55.67%) was obtained. 1 H NMR (300MHz, DMSO-d6) δ7.36 (ddt, J=7.6, 2.0, 1.0Hz, 1H), 7.23 (td, J=7.4, 2.0Hz, 1H ),7.09–7.01(m,2H),6.90(d,J=2.0Hz,1H),6.83(dd,J=7.4,1.9Hz,2H),6.58(s,2H), 5.99(s,1H),5.73(d,J=12.4Hz,1H),5.48(d,J=12.5Hz,1H),5.38(d,J=12.4Hz,1H), 5.13(d,J=12.5Hz,1H),5.10(d,J=0.9Hz,2H),4.51(s,2H),3.95(s,6H),3.84(s,3H). 13C NMR(100MHz,DMSO-d6)δ164.57,162.04,150.58,150.47,138.60,135.75,133.64,133.20,133.17,130.99,1 27.34,125.03,124.52,118.84,115.02,114.23,113.19,112.50,101.89,78.13,75.89,56.50,55.14.ESI-MS m / z:434.29[M+H] + .

[0098] Embodiment 11

[0099] Compound 11: 2,6-dimethoxy-4-{1-[4-methoxy-3-({[3-(prop-2-enyloxy)phenyl]methyl}oxy)phenyl]vinyl}pyridine

[0100] Using 3-hydroxymethylphenol and 3-bromopropylene as raw materials and following the procedure of Example 7, a white solid (79 mg, 52.36%) was obtained. 1 H NMR(300MHz, DMSO-d6)δ7.30–7.19(m,2H),7.06(dd,J=7.5,2.0Hz,1H),6.93(d,J=2.0Hz, 1H),6.88(dt,J=7.5,2.1Hz,1H),6.83(d,J=7.5Hz,1H),6.77(tt,J=2.1,0.9Hz,1H),6.58 (s,2H),5.99(s,1H),5.73(d,J=12.4Hz,1H),5.48(d,J=12.5Hz,1H),5.38(d,J=12.4Hz,1 H),5.13(d,J=12.5Hz,1H),5.02(t,J=1.0Hz,2H),4.50(s,2H),3.95(s,6H),3.84(s,3H). 13 C NMR(100MHz,DMSO-d6)δ164.57,158.12,150.95,150.42,138.60,136.14,135.75,133.20,133.16,131.04,1 25.98,124.52,118.84,118.06,116.48,115.02,113.19,111.91,101.89,80.07,75.71,56.50,55.19.ESI-MS m / z:434.35[M+H] + .

[0101] Example 12

[0102] Compound 12: 2,6-dimethoxy-4-{1-[4-methoxy-3-({[4-(prop-2-enyloxy)phenyl]methyl}oxy)phenyl]vinyl}pyridine

[0103] Using 4-hydroxymethylphenol and 3-bromopropylene as raw materials and following the procedure of Example 7, a white solid (91 mg, 60.31%) was obtained. 1 H NMR (300MHz, DMSO-d6) δ7.27(p,J=1.5Hz,1H),7.25(q,J=1.3Hz,1H),7.06(dd,J=7.5,2.0Hz,1H),6.93(d,J=2.0Hz,1H),6.85–6.83(m,3H),6.58(s,2H ),5.99(s,1H),5.73(d,J=12.4Hz,1H),5.48(d,J=12.5Hz,1H),5.38(dd,J= 12.4,0.0Hz,1H),5.13–5.11(m,3H),4.49(s,2H),3.95(s,6H),3.84(s,3H). 13 C NMR(100MHz,DMSO-d6)δ164.57,162.96,151.45,150.27,138.60,135.75,134.34,133.20,133.16,1 33.06,124.52,118.84,116.63,115.02,113.19,111.91,101.89,81.20,74.53,56.50,55.14.ESI-MS m / z:340.25[M+H] + .

[0104] Example 13

[0105] Compound 13: 2,6-dimethoxy-4-[1-(4-methoxy-3-{[(4-methoxyphenyl)methyl]oxy}phenyl)vinyl]pyridine

[0106] Using 5-[1-(2,6-dimethoxypyridin-4-yl)vinyl]-2-methoxyphenol, p-methoxybenzyl bromide and anhydrous potassium carbonate as raw materials, the reaction was carried out according to step 10 of Example 1 to obtain a white solid (99 mg, 69.81%). 1H NMR (300MHz, DMSO-d6) δ7.30–7.27(m,3H),7.06(dd,J=7.5,2.0Hz,1H),6.93(d,J=2.0Hz,1H),6.90–6.86(m,2H),6.83(d,J=7.5H z,1H),6.58(s,2H),5.73(d,J=12.4Hz,1H),5.48(d,J=12.4Hz,1H),5.11(t,J=1.0Hz,3H),3.95(s,6H),3.84(s,3H),3.79(s,3H). 13 C NMR(100MHz,DMSO-d6)δδ164.57,162.15,151.45,150.27,138.60,135.75,134.34,133.20, 132.87,124.52,115.64,115.02,113.19,111.91,101.89,81.20,56.50,56.03,55.1.ESI-MS m / z:408.19[M+H] + .

[0107] Embodiment 14

[0108] Compound 14: 2,6-dimethoxy-4-[1-(4-methoxy-3-{[(4-methylphenyl)methyl]oxy}phenyl)vinyl]pyridine

[0109] Using 5-[1-(2,6-dimethoxypyridin-4-yl)vinyl]-2-methoxyphenol, p-methylbenzyl bromide and anhydrous potassium carbonate as raw materials, the reaction was carried out according to step 10 of Example 1 to obtain a white solid (94 mg, 68.99%). 1 H NMR (300MHz, DMSO-d6) δ7.28–7.22(m,2H),7.15–7.04(m,3H),6.93(d,J=2.0Hz,1H),6.83(d,J=7.5Hz,1H),6.58(s ,2H),5.73(d,J=12.5Hz,1H),5.48(d,J=12.5Hz,1H),5.10(t,J=1.0Hz,2H),3.95(s,6H),3.84(s,3H),2.37(s,3H). 13C NMR(100MHz,DMSO-d6)δ164.57,151.45,150.23,141.29,138.89,138.60,135.75,133.20,1 31.29,130.45,124.52,115.02,113.19,111.91,101.89,81.68,56.50,55.14,22.32.ESI-MS m / z:317.27[M+H] + .

[0110] Embodiment 15

[0111] Compound 15: 2,6-dimethoxy-4-[1-(4-methoxy-3-{[(4-nitrophenyl)methyl]oxy}phenyl)vinyl]pyridine

[0112] Using 5-[1-(2,6-dimethoxypyridin-4-yl)vinyl]-2-methoxyphenol, p-nitrobenzyl bromide and anhydrous potassium carbonate as raw materials, the reaction was carried out according to step 10 of Example 1 to obtain a yellow powdery solid (62 mg, 42.17%). 1 H NMR (300MHz, DMSO-d6) δ8.12(d,J=1.6Hz,1H),8.10(d,J=1.6Hz,1H),7.61(p,J=1.4Hz,1H),7.59(q,J=1.2Hz,1H),7.06(dd,J=7.5,2.0Hz,1H),6.93( d,J=2.0Hz,1H),6.83(d,J=7.5Hz,1H),6.58(s,2H),5.73(d,J=12.4Hz,1H ),5.48(d,J=12.5Hz,1H),5.11(d,J=0.9Hz,2H),3.95(s,6H),3.84(s,3H). 13 CNMR(100MHz,DMSO-d6)δ164.57,151.45,150.45,149.29,142.24,138.60,135.75,133.2 0,131.03,125.27,124.52,115.02,113.19,111.91,101.89,81.39,56.50,55.14.ESI-MS m / z:423.51[M+H] + .

[0113] Example 16

[0114] Compound 16: 4-[({5-[1-(2,6-dimethoxypyridin-4-yl)vinyl]-2-methoxyphenyl}oxy)methyl]benzene-1-carbonitrile

[0115] Using 5-[1-(2,6-dimethoxypyridin-4-yl)vinyl]-2-methoxyphenol, 4-(bromomethyl)benzene-1-carbonitrile and anhydrous potassium carbonate as raw materials, the reaction was carried out according to step 10 of Example 1 to obtain a white solid (101 mg, 72.11%). 1 H NMR (300MHz, DMSO-d6) δ7.60(d,J=1.6Hz,1H),7.59(d,J=1.6Hz,1H),7.41(q,J=1.3Hz,1H),7.40(dt,J=2.0,1.0Hz,1H),7.06(dd,J=7.5,2.0Hz,1H),6. 93(d,J=2.0Hz,1H),6.83(d,J=7.5Hz,1H),6.58(s,2H),5.73(d,J=12.4Hz,1 H), 5.48 (d, J = 12.4Hz, 1H), 5.12 (t, J = 1.0Hz, 2H), 3.95 (s, 6H), 3.84 (s, 3H). 13 CNMR(100MHz,DMSO-d6)δ164.57,151.45,150.27,138.60,137.65,135.75,133.20,133.05,1 31.52,124.52,121.03,115.22,115.02,113.19,111.91,101.89,80.22,56.50,55.14.ESI-MS m / z:403.16[M+H] + .

[0116] Embodiment 17

[0117] The compound of the present invention is prepared into tablets:

[0118] tablet

[0119]

[0120] The above formulation is prepared into tablets by conventional methods.

[0121] Embodiment 18

[0122] Antiproliferation experiments were performed on the compounds of the present invention:

[0123] Experimental method: Cells were digested, counted, and prepared into a concentration of 3.5×10 4Cell suspension of cells / mL was added to each well of a 96-well plate, 100 μl of the cell suspension was added; the 96-well plate was placed in a 37°C, 5% CO2 incubator for 24 hours; the drug was diluted to the required concentration with complete culture medium, 100 μL of the corresponding drug-containing culture medium was added to each well, and a negative control group was set up at the same time; the 96-well plate was placed in a 37°C, 5% CO2 incubator for 72 hours.

[0124] CCK-8 method:

[0125] 1) Stain the 96-well plate with CCK-8, λ=450nm, and measure the OD value;

[0126] 2) Add 10 μL CCK-8 to each well and continue culturing in the incubator for 2-3 hours;

[0127] 3) Mix gently on a shaker for 10 minutes; read the OD value of each well using an enzyme-labeled instrument at λ=450 nm and calculate the inhibition rate.

[0128]

[0129] Table 1. IC values ​​of the compounds of the present invention for antiproliferative activity against human cancer cell lines and human normal cell lines 50 (nM)

[0130] Example HCT-8 K562 HepG2 MDA-MB-231 HFL-1 Mother drug 86 28 39 84 792 1 D C C E F 2 C C D F F 3 A C A B F 4 E D D F D 5 C D C D F 6 B D D E E 7 D A B C E 8 C E B D F 9 A A B A E 10 C A A C C 11 C C D B E 12 A C D F A 13 B B D E F 14 D C D E E 15 B A C A B 16 B B B A A

[0131] Note: Grade A means 1μM<IC 50 ≤3μM, Grade B means 3μM<IC 50 ≤6μM, C grade means 6μM<IC 50 ≤10μM, D grade means 10μM<IC 50 ≤20μM, E grade means 20μM<IC 50 ≤30μM, F grade means IC 50 >30μM.

[0132] The results are shown in Table 1. For tumor cell lines, the IC values ​​of the compounds of the present invention are 50 were greater than 1 μM, and their cytotoxicity was significantly reduced. 50 The lowest has exceeded 10 times. For normal cell lines, its safety has also been improved to a certain extent. 50 The results are generally more than 2 times, most of them reach 10 times, and some of them reach more than 20 times. The above results show that the bioorthogonal prodrug small molecule of the present invention has less toxic side effects, can effectively increase the therapeutic window, and improve safety.

[0133] Embodiment 19

[0134] The growth inhibition experiment of K562 cells after palladium resin catalytic release was carried out on the compounds of the present invention:

[0135] Experimental method: The compound was prepared into a 10mM concentration stock solution according to the corresponding requirements, and diluted to the final concentration according to the multiple for subsequent experiments. Different concentrations of bioorthogonal prodrug compounds and different doses of palladium resin were incubated with K562 cells in a 96-well plate for 72 hours, and then the inhibition rate was measured and calculated using the CCK-8 method.

[0136] Table 2. Experimental results of the inhibition of K562 cell growth after the compounds of the present invention were released by palladium resin

[0137]

[0138]

[0139] As can be seen from Table 2, after co-incubation with palladium resin, the killing effect of some bioorthogonal prodrug compounds of the present invention on tumor cells was restored, indicating that these bioorthogonal prodrug compounds can release active ingredients and effectively inhibit the growth of tumor cells under the activation of palladium resin, playing a therapeutic role with high efficiency and low toxicity.

[0140] Embodiment 20

[0141] In vivo safety evaluation experiments were conducted on the compounds of the present invention:

[0142] Experimental methods: 21 female ICR mice aged 4 weeks were provided by Shanghai Slake Experimental Animal Co., Ltd. The animals were randomly divided into blank control group, mother drug group, and prodrug group with 3 mice in each group. The blank control group was injected with the solvent DMF: Tween 80: 9% saline = 10:2:88 (V:V:V), and the mother drug group and prodrug group were injected with tail vein doses of 40 mg / kg, 80 mg / kg, and 160 mg / kg, respectively. The solvent was DMF: Tween 80: 9% saline = 10:2:88 (V:V:V). The drug was administered continuously for 21 days, and the body weight was measured every two days. *P<0.05;

[0143] **P<0.01; ***P<0.001 compared with the blank group.

[0144] Depend on Figure 2 It can be seen that some compounds in the present invention have no significant effect on the body weight of mice at concentrations higher than the effective dose. However, the original drug significantly reduces the body weight of mice under therapeutic concentration conditions. Therefore, compared with the original drug, the bioorthogonal prodrug molecules in the present invention have better safety in vivo.

[0145] Embodiment 21

[0146] In vivo anti-tumor experiments were conducted on the compounds of the present invention:

[0147] Experimental method: 24 female ICR mice aged 4 weeks were provided by Shanghai Slake Experimental Animal Co., Ltd. The cultured mouse liver cancer cell suspension H22 was collected at a concentration of 1X10 7 Each mouse was inoculated with 0.1 ml of the solution into the subcutaneous tissue of the right forelimb of the mouse. The diameter of the transplanted tumor was measured with a vernier caliper. The tumor grew to 100 mm 3 Around 24 hours later, the animals were randomly divided into groups, with 6 animals in each group, namely blank control group, palladium resin control group, mother drug group, and prodrug + palladium resin group. The blank control was injected with the solvent DMF: Tween 80: 9% saline = 10:2:88 (V:V:V), and the palladium resin control group and prodrug + palladium resin group were injected with Pd resin (1 mg in 50 μL of PBS) intratumorally before administration. The blank control group and the palladium resin control group were not given any drug. The mother drug group and the prodrug + palladium resin group were administered with a tail vein dose of 20 mg / kg, and the solvent was DMF: Tween 80: 9% saline.

[0148] =10:2:88 (V:V:V), and the drug was administered continuously for 21 days. After 21 days of administration, the mice were killed, and the tumors were removed by surgery and weighed. The tumor growth inhibition rate (%) was calculated, and the results were analyzed using SPSS17.0. The t-test was used for statistical analysis between the groups, and the calculation formula is as follows:

[0149]

[0150] Table 3. In vivo antitumor activity of the compounds of the present invention

[0151]

[0152] The results are shown in Table 3. The prodrug compounds of the present invention can produce higher concentrations of original drug molecules at the same dose, and thus have a stronger effect on tumor growth in vivo.

[0153] Therefore, the vinylpyridine prodrug compound of the present invention specifically activates the prodrug through a palladium resin catalytic reaction to target tubulin and inhibit the microtubule polymerization of tumor cells, releases the active ingredient under the activation of the palladium resin, and effectively inhibits the growth of tumor cells, and has high safety, small toxic side effects, can effectively increase the treatment window, play a high-efficiency and low-toxicity therapeutic effect, and improve the selectivity and accuracy of the treatment. The present invention can not only reduce the toxicity to normal cells, but also achieve efficient and accurate treatment at the tumor site, and has the potential for wide application.

Claims

1. A vinylpyridine prodrug compound, characterized in that: The structure of the compound is shown in Formula I: wherein R1, R2, R4, and R5 are each independently selected from hydrogen, halogen, hydroxyl, thiol, cyano, nitro, carboxyl, ester, acyl, amide, hydroxymethyl, aldehyde, sulfonyl, C1-C8 alkyl, C2-C8 alkenyl, C2-C8 alkynyl, C1-C4 alkoxy, C3-C6 cycloalkyl, C1-C4 alkylamino, C1-C4 imino, C6-C 10 Aryl, C3-C 10 Heteroaryl, C4-C8 heterocyclic group; R3 is selected from hydrogen, C1-C8 alkyl, C2-C8 alkenyl, C2-C8 alkynyl, C3-C8 cycloalkyl, C6-C 10 Aryl, C3-C 10 Heteroaryl, C4-C8 heterocyclic group.

2. The compound according to claim 1, characterized in that R1, R2, R4, and R5 are each independently selected from hydrogen, halogen, hydroxyl, acyl, amide, hydroxymethyl, C1-C8 alkyl, C2-C8 alkenyl, C2-C8 alkynyl, C1-C4 alkoxy, C3-C6 cycloalkyl, C1-C4 alkylamino, C6-C 10 Aryl, C3-C 10 Heteroaryl, C4-C8 heterocyclic group; R3 is selected from hydrogen, C1-C8 alkyl, C2-C8 alkenyl, C2-C8 alkynyl, C3-C8 cycloalkyl, C6-C 10 Aryl, C3-C 10 Heteroaryl, C4-C8 heterocyclic group.

3. The compound according to claim 1, characterized in that R1, R2, R4, R5 are each independently selected from hydrogen, halogen, C1-C4 alkoxy, C1-C4 alkylamino, C4-C8 heterocyclyl; R3 is selected from hydrogen, C1-C8 alkyl, C2-C8 alkenyl, C2-C8 alkynyl, C3-C8 cycloalkyl, C6-C 10 Aryl, C3-C 10 Heteroaryl.

4. The compound according to claim 1, characterized in that R1, R2 are selected from hydrogen, fluorine, chlorine, bromine, methoxy, ethoxy, methylamino, ethylamino, N,N-dimethylamino, tetrahydropyrrolyl; R3 is selected from hydrogen, methyl, ethyl, allyl, propargyl, 1-butyn-3-yl, 3-methyl-1-butyn-3-yl, 1-buten-3-yl, benzyl, 4-propargyloxybenzyl, 3-propargyloxybenzyl, 2-propargyloxybenzyl, 4-allyloxybenzyl, 3-allyloxybenzyl, 2-allyloxybenzyl, 4-nitrobenzyl, 4-methoxybenzyl, 4-cyanobenzyl, 4-methylbenzyl; R4 and R5 are selected from hydrogen, hydroxy, methoxy, ethoxy, methylamino, and N,N-dimethylamino.

5. The compound according to claim 1, characterized in that The compounds also include pharmaceutically acceptable salts.

6. A method for preparing the vinylpyridine prodrug compound according to claim 1, characterized in that: The following steps are involved: 2,6-dichloroisonicotinic acid is used as a raw material, a condensing agent is added, and it is condensed with N,O-dimethylhydroxylamine hydrochloride to obtain Weinreb amide, which is then reduced to obtain ketone iii, and the carbonyl group is protected by 2-bromoethanol, and the chlorine atom is substituted by different groups to obtain intermediate vi, and intermediate vii is obtained after deprotection under acidic conditions; intermediate vii is reacted with p-toluenesulfonyl hydrazide to obtain intermediate viii; halogenated phenols with different substitutions are protected by tert-butyldimethylchlorosilane to obtain intermediate x; intermediate viii and intermediate x are subjected to Heck reaction, and then the protecting group is removed to obtain the original drug xii; finally, different halogenated hydrocarbons are used to perform nucleophilic substitution on the hydroxyl group to obtain a vinyl pyridine prodrug such as a compound shown in formula I.

7. The vinylpyridine prodrug compound according to claim 1, characterized in that: Any one selected from the following compounds 1 to 16:

8. A pharmaceutical composition, characterized in that It comprises the compound represented by formula I in claim 1 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

9. Use of the vinylpyridine prodrug compound according to claim 1 or the pharmaceutical composition according to claim 8 in the preparation of a microtubule protein inhibitor.

10. Use of the vinylpyridine prodrug compound according to claim 1 or the pharmaceutical composition according to claim 8 in the preparation of anti-tumor drugs.

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