Benzodioxolane and pomalidomide combined compound and preparation method and application thereof

Abstract

The invention discloses a benzodioxolame and pomalidomide combined compound as well as a preparation method and application thereof. The structural formula of the compound is as shown in formula I in the specification. The compound disclosed by the invention has good tumor inhibition activity and relatively high application value.

Description

Technical Field

[0001] This invention relates to a compound, and more particularly to a compound of benzodioxolane bound to pomalidomide, its preparation method, and its application. Background Technology

[0002] Cancer poses a serious threat to human health. Existing chemotherapy drugs generally suffer from significant toxic side effects and are prone to drug resistance, necessitating the development of novel, highly effective, and low-toxicity antitumor compounds. Benzodioxane compounds are widely found in various active ingredients of traditional Chinese medicine (such as piperine and berberine), and studies have shown that they possess significant anti-inflammatory, antioxidant, and antitumor activities, inhibiting tumor cell proliferation by regulating pathways such as NF-κB and PI3K / Akt. However, natural benzodioxane compounds suffer from low bioavailability and insufficient targeting, requiring structural modification to optimize their pharmacological properties.

[0003]

[0004] Pomalidomide, a third-generation immunomodulatory agent (IMiD), exerts its therapeutic effect on multiple myeloma by inducing the degradation of IKZF1 / 3 through binding to CRBN protein. However, its monotherapy efficacy is limited, and it is not effective against solid tumors. Recent studies have found that the combination of benzodioxanone derivatives and IMiDs can synergistically enhance the antitumor effect, suggesting that structural fusion of the two may produce superior activity. Summary of the Invention

[0005] Objectives of the Invention: The first objective of this invention is to provide a compound of benzodioxolane bound to pomalidomide; the second objective of this invention is to provide a method for preparing the benzodioxolane bound to pomalidomide compound; and the third objective of this invention is to provide applications of the benzodioxolane bound to pomalidomide compound.

[0006] Technical solution: The benzodioxolane-bound pomalidomide compound of the present invention has the following structural formula:

[0007]

[0008] Among them, Linker is Where n is 2 to 6.

[0009] Preferably, the structural formula of the benzodioxolane-compounded pomalidomide compound is:

[0010] The preparation method of benzodioxane-bound pomalidomide according to the present invention includes the following steps:

[0011] (1) Compound 1 and Compound 2 were amidated to give intermediate 3;

[0012] (2) Intermediate 3 and the tert-butyloxycarbonyl-protected linker are coupled to obtain intermediate 4;

[0013] (3) Intermediate 4 is hydrolyzed to obtain intermediate 5;

[0014] (4) Intermediate 5 and compound 6 are reacted with amidation under alkaline conditions to obtain the target product.

[0015]

[0016] Where n is 2 to 6.

[0017] Preferably, in steps (1) and (4), the amidation reaction temperature is 10–40 °C.

[0018] Preferably, in steps (1) and (4), the condensing agent for the amidation reaction is 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethylurea hexafluorophosphate (HATU), benzotriazole-N,N,N′,N′-tetramethylurea hexafluorophosphate (HBTU), 1-butylphosphonic anhydride (T4P), O-benzotriazole-N,N,N′,N′-tetramethylurea tetrafluoroborate (TBTU), tetramethylchlorourea hexafluorophosphate (TCFH), carbonyl diimidazole (CDI), or (1-cyano The reaction mixture is 2-ethoxy-2-oxoethyleneaminooxy)dimethylamino-morpholine-carbomony hexafluorophosphate (COMU); preferably TCFH and T4P; the solvent for the reaction is dimethyl sulfoxide, N,N-dimethylformamide, dichloromethane, acetonitrile, anhydrous ethanol, preferably acetonitrile and dichloromethane; the base for the reaction is potassium carbonate, cesium carbonate, triethylamine, N,N-diisopropylethylamine (DIPEA), N-methylimidazole (NMI), preferably N,N-diisopropylethylamine (DIPEA) and N-methylimidazole (NMI).

[0019] Preferably, in step (2), the coupling reaction catalyst is Pd(OAc)2, Cs2CO3 and 4,5-bisdiphenylphosphine-9,9-dimethyloxane, the reaction solvent is 1,4-dioxane, and the reaction temperature is 90-110℃.

[0020] Preferably, in step (3), the acidic reagent is an ethyl hydrochloride solution, a trifluoroacetic acid dichloromethane solution, or an oxaloyl chloride methanol solution, with an ethyl hydrochloride solution being preferred.

[0021] The tautomers, optical isomers, deuterated derivatives, nitrogen oxides, solvates, pharmaceutically acceptable salts, or prodrugs of the benzodioxolane-bound pomalidomide compound described in this invention.

[0022] Salts of the compounds in this invention preferably comprise pharmaceutically acceptable salts of the compounds, said salts being prepared by any suitable method provided in the literature, for example, using inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, and phosphoric acid; or using organic acids such as acetic acid, maleic acid, succinic acid, mandelic acid, fumaric acid, malonic acid, pyruvic acid, oxalic acid, glycolic acid, and salicylic acid; pyranonic acids such as glucuronic acid and galacturonic acid; α-hydroxy acids such as citric acid and tartaric acid; amino acids such as aspartic acid and glutamic acid; aromatic acids such as benzoic acid and cinnamic acid; and sulfonic acids such as p-toluenesulfonic acid and ethanesulfonic acid.

[0023] In this invention, a pharmaceutically acceptable carrier is used, which is relatively non-toxic and harmless to the patient at a concentration consistent with the effective activity of the active ingredient, such that any side effects caused by the carrier do not negate the beneficial effects of the active ingredient. The pharmaceutically effective amount of the compound or a pharmaceutically acceptable salt thereof is preferably an amount that results in or affects the specific condition being treated. The compound of this invention, together with a pharmaceutically acceptable carrier known in the art, can be administered orally, parenterally, topically, nasally, ocularly, sublingually, rectally, vaginally, etc., using any effective conventional dosage form, including immediate-release, sustained-release, and time-release formulations.

[0024] The application of the benzodioxolane-conjugated pomalidomide compound described in this invention in the preparation of drugs for treating tumors.

[0025] The tumors are cervical cancer and breast cancer.

[0026] Beneficial effects: Compared with the prior art, the present invention has the following significant advantages: The present invention has compounds that link benzodioxane and pomalidomide through a series of flexible hydrazine linker structures, which have good antitumor activity and show better antitumor activity than piperine. Detailed Implementation

[0027] The technical solution of the present invention will be further described below with reference to the embodiments.

[0028] Example 1

[0029] The compound of benzo[d][1,3]dioxane and pomalidomide of the present invention has the chemical name (E)-3-(6-((2-(2-((2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl)amino)acetamido)ethyl)amino)benzo[d][1,3]dioxanepentanol-5-yl)-N-(4-(4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)acrylamide, and its structural formula is as follows:

[0030]

[0031] Its preparation method includes the following steps:

[0032] (1) Synthesis of (E)-3-(6-bromobenzo[d][1,3]dioxolane-5-yl)-N-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)aniline)acrylamide, with the following structural formula:

[0033]

[0034] Add 200 mg (0.74 mmol) of 3-(6-bromobenzo[d][1,3]dioxolane-5-yl)acrylic acid, 384 mg (0.81 mmol) of [(ethylcyano-2-oxoethyleneaminooxy)dimethylamino-morpholino-carbomony hexafluorophosphate] (COMU), 224 mg (2.21 mmol) of N-methylmorpholine (NMM), and 161 mg (0.59 mmol) of 4-(4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)aniline to a 100 mL round-bottom flask, dissolve in 2 mL of N,N-dimethylformamide (DMF), and stir at 15–25 °C for 5 h. The reaction solution was extracted with ethyl acetate (50 mL × 3), washed three times with saturated sodium chloride aqueous solution (50 mL × 3), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and separated by silica gel column chromatography (dichloromethane:methanol = 30:1) to obtain 158 mg of white solid, yield 40.6%.

[0035] (2) Synthesis of (E)-3-(6-((2-tert-butoxycarbonylaminoethyl)amino)benzo[d][1,3]dioxolane-5-yl)-N-(4-(4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)acrylamide, with the following structural formula:

[0036]

[0037] (E)-3-(6-bromobenzo[d][1,3]dioxolane-5-yl)-N-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)aniline)acrylamide 400 mg (0.76 mmol), N-Boc-1,2-diaminoethane 146 mg (0.91 mmol), 4,5-bis(diphenylphosphine)-9,9-dimethyloxanthracene 220 mg (0.38 mmol), palladium acetate 43 mg (0.19 mmol), and cesium carbonate 743 mg (2.28 mmol) were added sequentially to a reaction flask. 10 mL of 1,4-dioxane was used as the reaction solvent. Under nitrogen protection, the mixture was heated to 110 °C and stirred under reflux for 6 h. The solvent was removed by concentration under reduced pressure, and the product was purified by silica gel column chromatography (eluent: dichloromethane:methanol = 30:1) to give 250 mg of the product, with a yield of 54.4%.

[0038] (3) Synthesis of (E)-3-(6-((2-aminoethyl)amino)benzo[d][1,3]dioxolane-5-yl)-N-(4-(4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)acrylamide, with the following structural formula:

[0039]

[0040] 10 mL of ethyl hydrochloride solution was added to the product of the previous step reaction, and the reaction was carried out at 15-25°C for 2 hours. The product was then concentrated under reduced pressure to obtain 250 mg of the target product, with a yield of 98.2%.

[0041] (4) Synthesis of (E)-3-(6-((2-(2-((2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindole-4-yl)amino)acetamido)ethyl)amino)benzo[d][1,3]dioxacyclopentanol-5-yl)-N-(4-(4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)acrylamide

[0042] Add 58 mg (0.17 mmol) of (2-(2,6-dioxo-3-piperidinyl)-1,3-dioxoisoindoline-4-yl)aminoacetic acid, 75 mg (0.17 mmol) of (ethylcyano-2-oxoethyleneaminooxy)dimethylamino-morpholino-carbomony hexafluorophosphate (COMU), 48 mg (0.47 mmol) of N-methylmorpholine (NMM), and 80 mg (0.16 mmol) of (E)-3-(6-((2-aminoethyl)amino)benzo[d][1,3]dioxolane-5-yl)-N-(4-(4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)acrylamide to a 100 mL round-bottom flask in 3 mL of acetonitrile. Stir the mixture at 15–25 °C for 4 h. The solution was concentrated under reduced pressure and purified by column chromatography (dichloromethane:methanol = 40:1) to give 39 mg of a yellow-green solid, with a yield of 30.1%.

[0043] 11H NMR (500 MHz, DMSO-d6) δ 10.68 (s, 1H), 8.30 (t, J = 5.7 Hz, 1H), 8.27 (d, J = 2.2 Hz, 1H), 7.98 (dd, J = 8.4, 2.2 Hz, 1H), 7.82 (d, J = 15.3 Hz, 1H), 7.65 (d, J = 8.6 Hz, 1H), 7.48 (dd, J = 8.5, 6.9 Hz, 1H), 7.08 - 7.02 (m, 1H), 7.01 (d, J = 7.0 Hz, 1H), 6.96 (s, 1H), 6.55 (d, J = 4.3 Hz, 2H), 6.52 (s, 1H), 6.48 (s, 1H), 5.94 (s, 2H), 5.90 (t, J = 5.5 Hz, 1H), 5.22 (dd, J = 13.1, 5.3 Hz, 1H), 4.28 (s, 2H), 3.63 (s, 2H), 3.24 (tt, J = 13.3, 6.8 Hz, 4H), 3.12 - 3.09 (m, 2H), 3.09 - 3.02 (m, 2H), 2.98 (s, 2H), 2.80 (ddd, J = 17.3, 4.4, 2.5 Hz, 2H), 2.70 (s, 3H), 2.69 - 2.61 (m, 2H), 2.14 - 1.94 (m, 2H). 13 13C NMR (125 MHz, DMSO-d6) δ 209.46, 203.41, 197.30, 193.12, 189.84, 188.20, 182.16, 179.85, 178.65, 175.37, 172.84, 169.78, 169.33, 167.77, 166.50, 165.60, 165.08, 155.76, 153.89, 151.36, 148.75, 147.28, 146.21, 139.29, 139.20, 136.14, 135.25, 133.83, 132.19, 128.01, 121.90, 111.46, 108.85, 105.57, 101.17, 90.20, 63.80, 60.52, 57.11, 57.02, 49.56, 48.36, 30.91, 22.04.

[0044] Example 2

[0045] The benzo[d][1,3]dioxacyclopentanol-5-yl]-N-(4-(4-methylpiperidin-1-yl)methyl)-3-(trifluoromethyl)phenyl)acrylamide of the present invention has the following structural formula:

[0046]

[0047] Its preparation method includes the following steps:

[0048] (1) Based on Example 2, in step (2), N-tert-butoxycarbonyl-1,2-ethylenediamine was replaced with N-tert-butoxycarbonyl-1,3-propanediamine, and the other conditions remained unchanged. Steps (1) and (3) were the same as in Example 1, and the intermediate (E)-3-(6-((3-aminopropyl)amino)benzo[d][1,3]dioxolane-5-yl)-N-(4-(4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)acrylamide was obtained, with the following structural formula:

[0049]

[0050] (4) Add 91 mg (0.27 mmol) of (2-(2,6-dioxo-3-piperidinyl)-1,3-dioxoisoindoline-4-yl)aminoacetic acid, 84 mg (0.31 mmol) of N,N,N′,N′-tetramethylchloroformamidine hexafluorophosphate (TCFH), 72 mg (0.87 mmol) of NMI, and 130 mg (0.25 mol) of ((E)-3-(6-((3-aminopropyl)amino)benzo[d][1,3]dioxolane-5-yl)-N-(4-(4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)acrylamide to a 100 mL round-bottom flask in 4 mL of acetonitrile and stir at room temperature for 4 h. After concentration under reduced pressure, column chromatography (dichloromethane:methanol = 40:1) yielded 35 mg of a yellow-green solid, with a yield of 16.8%.

[0051] 11H NMR (400 MHz, DMSO-d6) δ 10.31 (s, 1H), 8.19 (d, J = 2.2 Hz, 1H), 8.05 - 8.01 (m, 1H), 7.99 (d, J = 6.0 Hz, 1H), 7.86 (dd, J = 8.4, 2.2 Hz, 1H), 7.80 (d, J = 15.1 Hz, 1H), 7.67 (s, 1H), 7.64 (s, 1H), 7.03 (s, 1H), 7.01 (s, 1H), 7.01 - 6.98 (m, 1H), 6.94 (s, 1H), 6.43 (d, J = 15.1 Hz, 1H), 6.37 (s, 1H), 5.92 (s, 2H), 5.70 (t, J = 5.7 Hz, 1H), 4.49 (s, 1H), 4.25 (s, 2H), 3.17 (d, J = 2.8 Hz, 2H), 3.05 (q, J = 6.2, 5.5 Hz, 4H), 2.91 - 2.69 (m, 4H), 2.64 (td, J = 13.4, 4.5 Hz, 4H), 2.28 (s, 3H), 2.12 - 2.08 (m, 1H), 2.07 (d, J = 5.8 Hz, 1H), 2.04 - 1.99 (m, 1H), 1.99 - 1.90 (m, 1H), 1.71 - 1.65 (m, 2H). 13 13C NMR (125 MHz, DMSO-d6) δ 171.82, 169.93, 168.97, 167.77, 166.63, 165.33, 151.00, 147.25, 145.16, 139.29, 139.21, 137.15, 135.95, 132.44, 131.85, 130.11, 122.21, 117.64, 116.44, 111.98, 111.48, 108.99, 101.21, 94.16, 57.75, 54.81, 54.80, 52.49, 52.48, 49.58, 49.06, 42.80, 40.65, 40.44, 40.23, 40.02, 39.81, 39.60, 39.39, 36.99, 27.06, 22.54.

[0052] Example 3

[0053]

[0054] The benzo[d][1,3]dioxapentanol-5-yl]-N-(4-(4-methylpiperidin-3-yl)-1,3-dioxoisoindole-4-yl)amino)acetamido)butyl)amino)benzo[d][1,3]dioxacyclopentanol-5-yl)-N-(4-(4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)acrylamide of the present invention has the following structural formula:

[0055] Its preparation method includes the following steps:

[0056] (1) Based on Example 2, in step (2), N-tert-butoxycarbonyl-1,2-ethylenediamine was replaced with N-tert-butoxycarbonyl-1,4-butanediamine, while the other conditions remained unchanged. Steps (1) and (3) were the same as in Example 1. The intermediate (E)-3-(6-((4-aminobutyl)amino)benzo[d][1,3]dioxolane-5-yl)-N-(4-(4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)acrylamide was obtained, with the following structural formula:

[0057]

[0058] (4) To a 100 mL round-bottom flask, add 137 mg (0.41 mmol) of (2-(2,6-dioxo-3-piperidinyl)-1,3-dioxoisoindoline-4-yl)aminoacetic acid, 126 mg (0.45 mmol) of N,N,N′,N′-tetramethylchloroformamidine hexafluorophosphate (TCFH), 108 mg (1.31 mmol) of NMI, and 200 mg (0.37 mol) of (E)-3-(6-((4-aminobutyl)amino)benzo[d][1,3]dioxolane-5-yl)-N-(4-(4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)acrylamide, dissolved in 4 mL of acetonitrile, and stirred at room temperature for 4 h. Concentrate under reduced pressure and precipitate by column chromatography (dichloromethane:methanol = 30:1) to obtain 30 mg of a yellow-green solid, yield 9.5%.

[0059] 1H NMR (500MHz, DMSO-d6) δ10.31 (s, 1H), 9.53 (s, 1H), 8.21 (d, J = 2.2Hz, 1H), 7.96 (t, J = 5.7Hz, 1H), 7.89 (dd, J = 8.5, 2.2Hz, 1H), 7.82 (d, J =15.1Hz, 1H), 7.66 (d, J = 8.5Hz, 1H), 7.47 (dd, J = 8.5, 6.9Hz, 2H), 7.21 (s, 1H), 7.10 (s, 1H), 7.04 (s, 1H), 6.95 (s, 1H), 6.38 (s, 1H), 5.9 4 (s, 2H), 5.76 (s, 1H), 5.33 (t, J=5.0Hz, 1H), 4.24 (s, 2H), 3.65 (s, 2H), 3.52 (d, J=1.7Hz, 2H), 3.18 (s, 2H), 3.14-3.08 (m, 2H), 2.97-2. 85 (m, 4H), 2.80 (s, 3H), 2.71-2.61 (m, 2H), 2.08 (dtd, J=12.4, 5.2, 2.6Hz, 2H), 2.05-1.92 (m, 2H), 1.57-1.53 ​​(m, 2H), 1.50-1.47 (m, 2H). 13 C NMR (125MHz, DMSO-d6) δ171.8, 169.9, 169.0, 167.8, 166.4, 165.4, 151.1, 15 0.0, 147.3, 142.5, 140.3, 139.6, 139.1, 137.4, 136.0, 132.4, 132.1, 130.0, 1 28.0, 124.6, 122.7, 122.2, 117.0, 116.5, 111.5, 109.0, 105.6, 101.2, 57.0, 53.3, 53.3, 49.8, 49.6, 49.1, 42.2, 37.4, 31.7, 30.31, 29.5, 27.3, 22.5, 21.0.

[0060] Example 4

[0061] The benzo[d][1,3]dioxolane and pomalidomide compounds of the present invention have the chemical name (E)-3-(6-((5-(2-((2-(2,6-dioxo-3-piperidine)-1,3-dioxoisoindoline-4-yl)amino)acetamido)pentyl)amino)benzo[d][1,3]dioxolane-5-yl-N-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)acrylamide, and the structural formula is as follows:

[0062]

[0063] Its preparation method includes the following steps:

[0064] (1) Based on Example 2, in step (2), N-tert-butoxycarbonyl-1,2-ethylenediamine was replaced with N-tert-butoxycarbonyl-1,5-pentanediamine, while the other conditions remained unchanged. Steps (1) and (3) were the same as in Example 2. The intermediate (E)-3-(6-((5-aminopentyl)amino)benzo[d][1,3]dioxolane-5-yl)-N-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)aniline)acrylamide was obtained, with the following structural formula:

[0065]

[0066] (4) 70 mg (0.21 mmol) of (2-(2,6-dioxo-3-piperidinyl)-1,3-dioxoisoindoline-4-yl)aminoacetic acid, 72 mg (0.25 mmol) of N,N,N′,N′-tetramethylchloroformamidin hexafluorophosphate (TCFH), and 61 mg (0.74 mmol) of N-methylimidazolium (NMI) were added sequentially to a reaction flask. 3 mL of acetonitrile was used as the reaction solvent. Then, (E... 128 mg (0.23 mmol) of 3-(6-((5-aminopentyl)amino)benzo[d][1,3]dioxolane-5-yl)-N-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)aniline)acrylamide was reacted at room temperature for 5 h. The solvent was removed by concentration under reduced pressure, and the product was purified by column chromatography (eluent: dichloromethane:methanol = 20:1) to give 20 mg of a yellow-green solid, with a yield of 11.1%.

[0067] 1H NMR (400MHz, DMSO-d6) δ10.30 (s, 1H), 8.19 (d, J = 2.2Hz, 1H), 7.92 (t, J = 5.6Hz, 1H), 7.88-7.76 (m, 2H), 7.65 (d, J = 8. 5Hz, 1H), 7.48 (dd, J=7.8, 3.8Hz, 1H), 7.01 (dd, J=10.2, 7.7Hz, 2H), 6.94 (s, 1H), 6.53 (s, 2H), 6.42 (d, J=15.1Hz, 1H) , 6.35 (s, 1H), 5.93 (s, 2H), 5.65 (s, 1H), 5.16 (dd, J = 13.0, 5.6Hz, 1H), 4.23 (s, 2H), 3.13-2.95 (m, 6H), 2.68 (s, 2H), 2.39 (d, J=23.3Hz, 8H), 2.23 (s, 5H), 1.56 (t, J=7.3Hz, 2H), 1.42 (q, J=7.1, 6.6Hz, 2H), 1.34 (dd, J=10.2, 5.5Hz, 2H). 13 C NMR (125MHz, DMSO-d6) δ171.70, 171.60, 169.82, 169.70, 168.91, 167.71, 166.31, 165.28, 150.92, 147.18, 145.30, 139.20, 139.06, 137.15, 135.88, 132.36, 131.76, 131.37, 126.11, 122.59, 122.14, 117.41, 116.8, 116.35, 111.79, 111.40, 108.92, 105.77, 101.12, 94.01, 57.73, 54.83, 52.60, 49.53, 45.59, 44.07, 42.64, 31.59, 29.30, 28.60, 24.34, 22.99, 21.65.

[0068] Example 5

[0069] The benzo[d][1,3]dioxanol-5-yl]acrylamide compound of the present invention has the chemical name (E)-3-(6-((6-(2-((2-(2-(2-(2-)dioxanol-3-yl)-1,3-dioxanol-4-yl)amino)acetamido)hexyl)amino)benzo[d][1,3]dioxanol-5-yl)-N-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)acrylamide, and its structural formula is as follows:

[0070]

[0071] Its preparation method includes the following steps:

[0072] (1) Based on Example 2, in step (2), N-tert-butoxycarbonyl-1,2-ethylenediamine was replaced with N-tert-butoxycarbonyl-1,6-hexanediamine, while the other conditions remained unchanged. Steps (1) and (3) were the same as in Example 2. The intermediate ((E)-3-(6-((6-aminohexyl)amino)benzo[d][1,3]dioxolane-5-yl)-N-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)aniline)acrylamide was obtained, with the following structural formula:

[0073]

[0074] (4) 99 mg (0.30 mmol) of (2-(2,6-dioxo-3-piperidinyl)-1,3-dioxoisoindoline-4-yl)aminoacetic acid, 100 mg (0.36 mmol) of N,N,N′,N′-tetramethylchloroformamidine hexafluorophosphate (TCFH), and 86 mg (1.04 mmol) of N-methylimidazolium (NMI) were added sequentially to a reaction flask. 3 mL of acetonitrile was used as the reaction solvent. Then, (E... 184 mg (0.33 mmol) of 3-(6-((6-aminohexyl)amino)benzo[d][1,3]dioxolane-5-yl)-N-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)aniline)acrylamide was reacted at room temperature for 5 hours. The solvent was removed by concentration under reduced pressure, and the mixture was purified by column chromatography (eluent: dichloromethane:methanol = 20:1) to give 70 mg of a deep yellow solid, with a yield of 26.7%.

[0075] 1H NMR (400MHz, DMSO-d6) δ10.33 (s, 1H), 8.20 (d, J = 2.2Hz, 1H), 7.92-7.86 (m, 2H), 7.80 (d, J = 15.2Hz, 1H), 7.65 (d, J = 8.5Hz, 1H ), 7.46 (dd, J=8.5, 7.0Hz, 1H), 7.01 (dd, J=9.9, 7.7Hz, 2H), 6.94 (s, 1H), 6.52 (s, 2H), 6.42 (d, J=15.1Hz, 1H), 6.35 (s, 1H), 5 .93 (s, 2H), 5.65 (s, 1H), 5.16 (dd, J=13.0, 5.3Hz, 1H), 4.22 (s, 2H), 3.63 (d, J=4.3Hz, 2H), 3.04 (tdd, J=17.3, 12.1, 6.3Hz, 1 0H), 2.76 (s, 6H), 2.69 (s, 3H), 2.08 (dt, J = 10.0, 2.7Hz, 2H), 1.54 (q, J = 7.3Hz, 2H), 1.40 (t, J = 6.9Hz, 2H), 1.33-1.25 (m, 2H). 13 C NMR (125MHz, DMSO-d6) δ171.49, 169.61, 168.70, 167.49, 166.05, 165.11, 162.53, 158.92, 15 0.76, 147.85, 146.96, 145.17, 141.09, 139.31, 138.86, 137.08, 135.68, 132.15, 131.80, 130 .15, 126.25, 122.37, 121.93, 117.06, 111.19, 108.69, 105.52, 100.92, 87.54, 80.97, 76.94, 56.76, 53.04, 49.62, 49.31, 43.86, 42.41, 34.52, 31.36, 29.25, 28.67, 26.55, 26.35, 21.42.

[0076] Effect Test Case

[0077] MTT assay for antitumor activity

[0078] Cell culture: Human cancer cell lines HeLa and MDA-MB-231 were purchased from the National Biomedical Laboratory in Beijing and cultured in DMEM (KGM12800-500) or MEM (KGM41500-500) medium containing 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin, in a Thermo Fisher Scientific, BB150 incubator at 37°C with 5% CO2. When the cell confluence reached 70%-80%, 0.25% trypsin was added for digestion, resuspending, and culturing. Cells in the logarithmic growth phase and in good growth status were selected for study.

[0079] Methyl thiazolyl tetrazolium (MTT) was used to determine cell viability. Hemocytometer counting was used for cell counting, and cell viability was greater than 95% in all experiments. MDA-MB-231 and HeLa cells were counted at 1 × 10⁻⁶ cells / mL. 4 Cells were seeded in 96-well plates. 100 μL of medium (containing 1% FBS) was added to each well to dissolve different concentrations of the drug (0-50 μM), and the cells were incubated for 24 h. After centrifugation (5 min, 2000 rpm), the supernatant was discarded, and 10 μL of LTT (5 mg / mL) solution was added to each well. The cells were incubated at 37°C for 4 h, centrifuged again, and the supernatant was discarded. 100 μL of DMSO was added to each well, and the cells were shaken for 10 min to fully dissolve the formazan crystals. The absorbance was measured at 570 nm using a microplate reader (BioTek, USA). Cytotoxicity was assessed compared to the control group (DMSO). The concentration at which the drug induced 50% cell growth inhibition (IC50) was determined using a curve fitting algorithm in GraphPad Prism 9 (GraphPad software, LaJolla, CA, USA) via nonlinear regression. 50 Table 1 shows the effects of the compounds on the activity of different tumor cells, and the half-maximal inhibitory concentration (IC50) of the drugs on each cell line was calculated. 50 (48h).

[0080] Table 1. Inhibitory activity of compounds against different tumor cells (IC50) s0 )

[0081] Example Hela (μM) MDA-MB-231(μM) Example 1 21.36±2.19 30.7±2.37 Example 2 / 63.31±2.51 Example 3 28.86±0.38 11.05±0.65 Example 4 9.43±0.38 3.12±0.32 Example 5 15.34±0.15 6.37±0.26 Example 6 / 39.88±15.49 Piperine 44.37±3.28 239.03±3.65 pomalidomide 572.73±13.28 389.47+35.83

[0082] " / " indicates that it was not measured.

[0083] The results in Table 1 show that these compounds have a significant inhibitory effect on HELA and MDA-MB-231 tumor cells, which is significantly better than that of piperine, a typical natural product containing benzodioxolane. This indicates that the compounds have good activity against tumor cells and have great research value.

Claims

1. A benzodioxolane-conjugate pomalidomide compound, characterized by, The chemical structural formula is: wherein Linker is wherein n is 2 to 6.

2. The benzodioxole-bound pomalidomide compound of claim 1, wherein, It is a compound as shown below or a pharmaceutically acceptable salt thereof:

3. A method of preparing the benzodioxolane-conjugated pomalidomide compound of claim 1, comprising: The method comprises the following steps: compound 5 and compound 6 are subjected to an amidation reaction to obtain compound I:

4. A compound as shown below: wherein, n is 2-6.

5. The compound of claim 4, which has the following structure:

6. A pharmaceutical composition, characterized by, It comprises the compound of claims 1-2 and at least one pharmaceutically acceptable excipient.

7. Use of a substance for the manufacture of a medicament for the treatment of a disease, characterized in that, The substance is the compound of claims 1-2, and the disease is cancer; preferably cervical cancer, breast cancer.

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