CN103965398B - D-semi-lactosi/phenyl aldehyde mustargen/N-(2-hydroxypropyl) methacrylamide copolymer and Synthesis and applications thereof - Google Patents

Abstract

The invention provides a kind of D-galactose/benzaldehyde nitrogen mustard/N-(2-hydroxypropyl) methacrylamide copolymer, is that D-galactose, benzaldehyde nitrogen mustard are connected to N by covalent bond -(2-hydroxypropyl)methacrylamide (HPMA) to form a polymer copolymer with good biocompatibility, which belongs to the field of polymer chemistry and application. Experiments have proved that the D-galactose/benzaldehyde nitrogen mustard/N-(2-hydroxypropyl) methacrylamide copolymer of the present invention has the function of targeted intelligent drug release, and has a good inhibitory effect on liver cancer HepG2 tumor cells effect; at the same time, the polymer is also an effective drug delivery system. Therefore, as an inhibitor of HepG2 tumor cells and the like, it has a good prospect in the preparation of antitumor drugs.

Description

D-galactose/benzaldehyde nitrogen mustard/N-(2-hydroxypropyl)methacrylamide copolymer and its preparation and application

technical field

The invention belongs to the field of macromolecular chemistry, and relates to a D-galactose/benzaldehyde nitrogen mustard/N-(2-hydroxypropyl)methacrylamide copolymer with targeted anti-tumor activity of intelligent drug release; At the same time, it also relates to a preparation method of the copolymer; the invention also relates to the application of the polymer as an inhibitor of liver cancer HepG2 cells in the preparation of anticancer drugs.

Background technique

Nitrogen mustard compounds are alkylating agent antineoplastic drugs, but due to their high toxicity and side effects, there are certain limitations in clinical application. Currently, they are mainly used in the treatment of malignant lymphomas. According to the parent structure, nitrogen mustards can be divided into benzoate mustard, chlorambucil, benzaldehyde mustard and methyl mustard. Wherein the structural formula of benzaldehyde nitrogen mustard is as follows:

Due to the poor water solubility of benzaldehyde nitrogen mustard, co-solvents (absolute ethanol, DMSO, etc.) need to be used during application, which will cause various toxic and side effects in the body. In addition, after being diluted with water, it needs to be filtered through a 0.25 μm microporous membrane, which makes the administration inconvenient.

D-galactose is a small molecular substance capable of targeting recognition and binding to the asialoglycoprotein receptor on the surface of liver cells, and its strong hydrophilicity can also increase the water solubility of the polymer. The structural formula of its D-galactose is as follows:

N-(2-hydroxypropyl)methacrylamide (HPMA) is a polymer drug carrier with good biocompatibility, which can not only reduce the side effects of drugs, reduce drug resistance, and improve the stability of drugs in vivo , can also increase the accumulation of drugs in the tumor site, so that the drug effect can be better exerted, etc., and has been used as a tumor-targeting drug carrier in clinical practice. Its structural formula is as follows:

Therefore, using N-(2-hydroxypropyl) methacrylamide with good water solubility as a drug carrier, a polymer copolymer with anti-tumor activity can be obtained by copolymerizing with double bond-containing D-galactose and benzaldehyde nitrogen mustard , which is expected to provide more choices for the anti-cancer field.

Contents of the invention

The purpose of this invention is to utilize the characteristics of benzaldehyde nitrogen mustard, D-galactose and N-(2-hydroxypropyl) methacrylamide to provide a kind of D-galactose/benzaldehyde nitrogen mustard/N-(2- Hydroxypropyl) methacrylamide copolymer.

Another object of the present invention is to provide a preparation method of D-galactose/benzaldehyde nitrogen mustard/N-(2-hydroxypropyl)methacrylamide copolymer.

Another object of the present invention is to provide the application of D-galactose/benzaldehyde nitrogen mustard/N-(2-hydroxypropyl)methacrylamide copolymer as an inhibitor of liver cancer HepG2 cells in the preparation of antitumor drugs.

(1) D-galactose/benzaldehyde nitrogen mustard/N-(2-hydroxypropyl)methacrylamide copolymer

D-galactose/benzaldehyde nitrogen mustard/N-(2-hydroxypropyl) methacrylamide copolymer of the present invention is that D-galactose and benzaldehyde nitrogen mustard are connected to N-(2- Hydroxypropyl) methacrylamide (HPMA) is a polymer copolymer with good biocompatibility. Its specific preparation process is as follows:

(1) compound Preparation: use tetrahydrofuran as a solvent, p-toluenesulfonic acid as a catalyst, and molecular sieves as a water-absorbing agent, under the protection of nitrogen, react benzaldehyde nitrogen mustard with 1.1.1-trimethylolethane at -5 ~ 5 ° C 12-15 hours; after the reaction of benzaldehyde nitrogen mustard is complete, add ammonia water to neutralize p-toluenesulfonic acid to pH=7-8, filter out molecular sieves, concentrate the filtrate, dilute with dichloromethane, buffer with pH=8 phosphate Liquid washing, drying over anhydrous sodium sulfate, separation by silica gel column chromatography, to obtain the compound . Its structural formula is:

The molar ratio of benzaldehyde nitrogen mustard to 1.1.1-trimethylolethane is 1:2.9～1:3.3; the molar ratio of benzaldehyde nitrogen mustard to catalyst p-toluenesulfonic acid is 1:0.05～1:0.1 The addition of molecular sieve is 15～20 times of the quality of benzaldehyde nitrogen mustard.

(2) compound Preparation: with dichloromethane as solvent, triethylamine as acid-binding agent, under nitrogen protection, the compound React with methacryloyl chloride at -5-0°C for 12-15 hours; after the reaction is complete, wash with saturated sodium bicarbonate, dry over anhydrous sodium sulfate, and separate by silica gel column chromatography to obtain the compound . Its structural formula is:

compound The molar ratio of methacryloyl chloride to methacryloyl chloride is 1:1～1:1.2; the molar ratio of methacryloyl chloride to acid-binding agent triethylamine is 1:1.1～1:1; the eluent used in silica gel column chromatography is: Ethyl acetate and petroleum ether in a volume ratio of 1:10 to 1:30.

(3) compound Preparation of galactose: Dissolve galactose in acetone, add concentrated sulfuric acid as a dehydrating agent, stir overnight at room temperature, until the solution turns bright yellow, add potassium carbonate solution to neutralize concentrated sulfuric acid, precipitate white solid, filter with suction, and distill out acetone , extracted with chloroform, washed with distilled water, and dried over anhydrous sodium sulfate to obtain a colorless transparent oil, which is the compound . Its structural formula is:

The amount of concentrated sulfuric acid added is 2% to 3% of the mass of galactose; the mass concentration of potassium carbonate solution is 15% to 20%; the eluent used in silica gel column chromatography is: ethyl acetate and petroleum ether at a ratio of 1:10 to 1 :30 volume ratio.

(4) compound Preparation: Compound Dissolve in anhydrous ether, add triethylamine, cool down to -5~5°C, add methacryloyl chloride, react for 3~12 hours; remove the precipitate by suction filtration, evaporate the filtrate to dryness, and dissolve the residue with n-hexane , remove the insoluble matter, distill out n-hexane to obtain a colorless viscous substance, which is the compound . Its structural formula is:

compound The molar ratio with methacryloyl chloride is 1:1～1:1.2; compound The molar ratio with triethylamine is 1:1.3～1:1.5; the amount of methacryloyl chloride added is the compound 0.4 to 0.6 times the mass.

(5) compound Preparation: Compound Dissolve in trifluoroacetic acid-water mixture, stir at room temperature to fully hydrolyze, then precipitate the hydrolyzate with ethanol, filter with suction to obtain a light yellow solid, recrystallize with ethanol to obtain a white solid which is the compound . Its structural formula is:

In the trifluoroacetic acid-water mixture, the volume ratio of trifluoroacetic acid to water is 9:1-8:1.

(6) Preparation of target compound: N-(2-hydroxypropyl)methacrylamide, compound and compounds Dissolve with DMSO and acetone, add initiator azobisisobutyronitrile, react at 50-60°C for 20-24 hours under nitrogen protection, precipitate with a mixture of acetone and ether, filter, dissolve the precipitate with anhydrous methanol, and finally use The ultrafiltration concentration centrifuge tube with a molecular weight of 3000 is centrifuged to remove small molecules to obtain the target high molecular weight copolymer. Its structure is as follows:

In the formula, x=80～90mol%, y=5～10mol%, z=5～10mol%; number average molecular weight Mn=2.1～2.8×10 ⁴ , Mw/Mn=1.33～1.81.

compound , compound , N-(2-hydroxypropyl) methacrylamide molar ratio is 1:1:4～1:1:20; The consumption of described initiator azobisisobutyronitrile is compound , compound and 5%-10% of the total mass of N-(2-hydroxypropyl)methacrylamide; in the mixture of acetone and ether, the volume ratio of acetone and ether is 7:3-7:1.

Figure 1 is the hydrogen nuclear magnetic resonance spectrum of the targeted anti-tumor active polymer with intelligent drug release prepared by the above method. Through proton nuclear magnetic resonance spectrum analysis, it can be concluded that the polymer obtained by copolymerization has a chemical shift between 6.5 and 7.5 ppm, and there is a peak of hydrogen on the aromatic ring in the compound benzaldehyde nitrogen mustard, and the chemical shift is at 3.70 ppm and 2.81 ppm. The characteristic peaks -CH- (connected to -OH) and -CH ₂ (connected to -NH-) on HPMA will appear, and the chemical shift is between 4.22ppm and 5.12ppm. The obtained product is consistent with the previously designed structure.

2. Antitumor activity experiment

1. In vitro tumor cell growth inhibition experiment

The HepG2 cell line was tested by the tetrazolium salt reduction method (MTT): take the liver cancer H22 cells in the logarithmic phase of growth, adjust the cell concentration to 2×10 ⁴ cells/ml, add 90 μl/ml to the 96-well culture plate Wells, edge wells were filled with sterile PBS. Incubate at 5% CO ₂ at 37°C, place in the incubator until adhered to the wall, and then add drugs. For the high molecular weight polymer of the present invention and benzaldehyde nitrogen mustard (4-di-(2-chloroethyl)benzaldehyde), 4 gradients of 1, 5, 25, and 125 μg/ml concentrations were respectively set. Both the experimental group and the control group had 4 duplicate wells. After adding the drug, the cells were cultured in a carbon dioxide incubator at a temperature of 37°C for 24 hours, and then they were taken out and centrifuged first, then discarded the supernatant culture solution in the 96-well plate, and carefully washed with PBS. After washing 2 to 3 times, add 20 μl of MTT (tetrazolium, 5 mg/ml, ie 0.5% MTT) solution to each well, and place it in a carbon dioxide incubator at 37°C to continue culturing for 4 hours. Terminate the culture, and carefully aspirate the culture medium in the well. Add 150 μgl of DMSO to each well, shake on a shaker at low speed for 10 minutes, and fully dissolve the crystals. The absorbance OD value of each well was measured in a microplate reader at 570 nm. The IC ₅₀ of the polymer was 11.7μg/ml, and the IC ₅₀ of benzaldehyde mustard was 25.9μg/mL. The cell growth inhibition rate was calculated according to the following formula:

Inhibition rate=[(1-average OD value of experimental group)/average OD value of control group]×100%

The test results are shown in Table 1 and Figure 2.

From the results of the above cell experiments, it can be seen that the polymer copolymer with targeting groups has a good inhibitory effect on HepG2 cells. On the one hand, pure benzaldehyde mustard has little solubility in aqueous solution, so it can hardly touch the surface of cells; on the other hand, pure benzaldehyde mustard has no targeting group and can only be passively taken up by cells , so it limited the inhibitory effect of benzaldehyde nitrogen mustard on HepG2 cells. And poly(HPMA-galactose-nitrogen mustard) is a water-soluble polymer compound, so it can accumulate on the surface of HepG2 cells and through the targeting group galactose on the polymer and the receptor overexpressed on the surface of HepG2 cells asialo The specific recognition of glycoprotein receptors is endocytosed by HepG2 cells under the mediation of the receptors, which increases the uptake of polymers by HepG2 cells, thus significantly enhancing the inhibitory effect on HepG2 cells. From the analysis of the results, the IC ₅₀ values of pure benzaldehyde nitrogen mustard and poly(HPMA-galactose-nitrogen mustard) on HepG2 cells were 25.9 μg/ml and IC ₅₀ =11.7 μg/ml, and poly(HPMA-galactose - nitrogen mustard) has an _IC50 value that is essentially half that of pure benzaldehyde nitrogen mustard.

2. In vitro simulated release experiment

In poly(HPMA-galactose-nitrogen mustard), benzaldehyde, a small molecule drug nitrogen mustard, is connected to the polymer chain through a pH-sensitive acetal bond. In the normal body fluid of the human body (pH=7.4), the acetal bond is stable, but in the tumor site (pH=4.5~6.9), the acetal bond will be hydrolyzed, thereby releasing the small molecule drug nitrogen mustard benzaldehyde . In order to test the release rate of the small molecule drug nitrogen mustard benzaldehyde in poly(HPMA-galactose-nitrogen mustard) under different pH conditions, the aqueous solution of poly(HPMA-galactose-nitrogen mustard) with a concentration of 0.2mg/mL The pH value was adjusted to 7.4, 5 and 4 by phosphate buffer solution and placed in a constant temperature water solution at 37°C, and the released nitrogen mustard benzaldehyde was extracted with chloroform at different times, and was measured at a wavelength of 312nm by an ultraviolet-visible spectrometer. Measure the absorbance of the extract, and finally add a drop of concentrated hydrochloric acid to completely hydrolyze the nitrogen mustard benzaldehyde, and use the ratio of the absorbance at different times to the total absorbance as the release rate at different times (see Figure 3). The results show that poly(HPMA-galactose-nitrogen mustard) releases nitrogen mustard benzaldehyde slowly in phosphate buffer at pH=7.4, and the release amount is close to 20% in 100 hours, while at pH=4 and 5 The release is fast, and there is an obvious burst release in the first 40 hours, and then the release rate is relatively slow, and the sustained release time is prolonged. The results show that poly(HPMA-galactose-nitrogen mustard) can exist stably in the buffer solution of pH=7.4, but it can release small molecule nitrogen mustard benzaldehyde under the conditions of pH=4 and 5, reaching the The purpose of slow release under different pH conditions.

In summary, the polymer copolymer of the present invention superimposes the antitumor activity of D-galactose, benzaldehyde nitrogen mustard and N-(2-hydroxypropyl) methacrylamide, and further promotes the antitumor activity of the polymer on tumors. Inhibitory effect, while greatly prolonging the residence time of anticancer drugs in tumors. Experiments have proved that D-galactose/benzaldehyde nitrogen mustard/N-(2-hydroxypropyl)methacrylamide copolymer of the present invention has the function of targeted intelligent drug release, and has a good effect on liver cancer HepG2 tumor cells. Inhibitory effect, and the polymer carrier polyHPMA also reduces the toxicity of anticancer drugs, and at the same time shows good biocompatibility, thereby reducing the damage to normal tissues; at the same time, the polymer is also an effective drug delivery system . Therefore, using the polymer as an inhibitor of HepG2 tumor cells in the preparation of antitumor drugs has a good prospect.

Description of drawings

Fig. 1 is the H NMR spectrum of the targeted anti-tumor active polymer with intelligent drug release in the present invention.

Fig. 2 is the 24h anticancer activity curve of the high molecular weight polymer of the present invention and benzaldehyde nitrogen mustard in vitro.

Fig. 3 is the relationship between the amount and time of nitrogen mustard benzaldehyde released from the targeted anti-tumor active polymer with intelligent drug release in the phosphate buffer solution with pH 7.4, 5 and 4 respectively.

detailed description

The synthesis and structural characterization of the polymer copolymer of the present invention will be further described below through specific examples.

Example 1

(1) compound Preparation: Dissolve benzaldehyde nitrogen mustard (1.0g, 5.6mmol, 1.0equiv.) in 75mL tetrahydrofuran, add 15-20g molecular sieves, and add p-toluenesulfonic acid (0.13g, 0.75 mmol, 0.13equiv.), 1,1,1-trimethylolethane (1.45g, 16mmol, 3.0equiv.), reacted in an ice-water bath for 12-15h; tracked the reaction with thin-layer chromatography, and waited for benzaldehyde nitrogen mustard After the reaction is complete, add ammonia water to neutralize p-toluenesulfonic acid to pH = 7-8, filter out molecular sieves, concentrate the filtrate, dilute with dichloromethane, wash three times with phosphate buffer solution with pH = 8, wash with anhydrous sulfuric acid Sodium drying for 12 hours, separated by silica gel column chromatography (ethyl acetate/petroleum ether=1/101 (v/v)), collecting product components, distilling off the solvent to obtain the compound (1.08g). Yield (78%).

¹ HNMR(400MHz, CDCl ₃ ): δ7.61(d, J =7.2Hz, 2H, –CHC ₆ H ₂ H ₂ N(CH ₂ CH ₂ Cl) ₂ ), 6.79(d, J =7.2Hz, 2H ,–CHC ₆ H ₂ H ₂ N(CH ₂ CH ₂ Cl) ₂ ),5.18(s,1H,–CHC ₆ H ₄ N(CH ₂ CH ₂ Cl) ₂ ),4.45(s,2H,– OCH ₂ C( CH ₃ )(CH ₂ O–) ₂ ),3.48-3.86(m,12H,–OCH ₂ C(CH ₃ )( CH ₂ O–) ₂ and–O(–O)CHC ₆ H ₄ N(CH _{2 CH 2} Cl _{) 2} )), 0.83(s,3H,–OCH ₂ C( CH ₃ )(CH ₂ O–) ₂ ).

(2) compound Preparation: take the compound (1.56g, 4.5mmol) and triethylamine (0.68g, 6.7mmol) were dissolved in 50mL of dichloromethane, cooled to about 0°C in an ice-water bath; methacryloyl chloride (0.56g, 5.4mmol) was dissolved in 10mL In dichloromethane, it was slowly added dropwise to the above solution, and reacted for 12 to 15 hours. The reaction was followed by TLC until the compound After the reaction, the reaction solution was washed three times with saturated sodium bicarbonate solution, dried with anhydrous sodium sulfate for 10 to 12 hours, filtered with suction, and the crude product was separated with a silica gel column after the filtrate was concentrated (ethyl acetate/petroleum ether=1/20 (v/ v)), the product components were collected, and the solvent was evaporated to obtain 1.05 g of a white solid with a yield of 56%.

¹ HNMR(400MHz,CDCl ₃ ,δ,ppm):7.61(d, J =7.2Hz,2H,–CHC ₆ H ₂ H ₂ N(CH ₂ CH ₂ Cl) ₂ ),6.79(d, J =7.2Hz ,2H,–CHC ₆ H ₂ H ₂ N(CH ₂ CH ₂ Cl) ₂ ),6.13(s,1H,–OC(CH ₃ )C=CH H ),5.57(s,1H,–OC(CH ₃ )C=C H H),5.18(s,1H,–CH C ₆ H ₄ N(CH ₂ CH ₂ Cl) ₂ ),4.45(s,2H,–OC H ₂ C(CH ₃ )(CH ₂ O–) ₂ ),3.48-3.86(m,12H,–OCH ₂ C(CH ₃ )( CH ₂ O–) ₂ and–O(–O)CHC ₆ H ₄ N(CH ₂ CH ₂ Cl ) ₂ )), 1.97(s,3H,–OC( CH ₃ )C=CH ₂ ), 0.83(s,3H,–OCH ₂ C( CH ₃ )(CH ₂ O–) ₂ ).

(3) compound Preparation: Weigh 1.0g (5.6mmol) of D-galactose and dissolve it in 50mL of acetone, add 0.5-1.2mL of concentrated sulfuric acid, stir the solution at room temperature overnight, the solution turns bright yellow, add 20% carbonic acid Potassium solution 10mL, a large amount of white solids were precipitated, suction filtered, acetone in the filtrate was distilled off, the remaining filtrate was extracted with 5×15ml of chloroform, then washed with 2×5ml of distilled water, and dried by adding anhydrous Na ₂ SO ₄ for 5 hours Afterwards, the chloroform was distilled off, and pumped with an oil pump for 20 minutes to obtain 1.2 g of a colorless and transparent oil, with a yield of 83%.

¹ HNMR (400MHz, CDCl ₃ , δ, ppm): 1.32–1.50(m,12H), 4.07(m,1H), 4.23–4.37(m,4H), 4.62(dd,1H), 5.53(d,1H ).

(4) compound Preparation: Take 1.2g (4.7mmol) of the compound Dissolve in 60mL of anhydrous ether, add 0.474g (4.7mmol) of triethylamine, cool to about 0°C in an ice-water bath, dissolve 0.489g (4.7mmol) of methacryloyl chloride in 30mL of ether, and separate The funnel was slowly added dropwise to the above solution, and then reacted for 3h after the drop was completed. The produced precipitate was filtered off with suction, the filtrate was evaporated to dryness, and 50 mL of n-hexane was added, and there was a small amount of insoluble matter. After removal, the n-hexane was evaporated to obtain 1.03 g of a colorless sticky substance, with a yield of 68%.

¹ HNMR (400MHz, CDCl ₃ , δ, ppm): 1.32–1.50(m,12H), 1.94(s,3H), 4.07(m,1H), 4.23–4.37(m,4H), 4.62(dd,1H ), 5.53(d,1H).5.56(s,1H),6.13(s,1H).

(5) compound Preparation: Take 1.03g (3.1mmol) of compound In a 50mL round bottom flask, add 5mL of a mixture of trifluoroacetic acid-water with a volume ratio of 9:1, stir at room temperature for 15min, add ethanol, and remove the solvent by suction filtration to obtain a pale yellow solid, which is recrystallized with ethanol to obtain White solid 0.68g, yield 87%.

¹ HNMR(400MHz,D ₂ O,δ,ppm):1.95(s,3H,–OC( CH ₃ )C=CH ₂ ),4.08,4.19,4.42,4.63,and5.04(7H,sugarmoiety), 6.13(s,1H,–OC(CH ₃ )C=CH H ),5.57(s,1H,–OC(CH ₃ )C=CH H ).

(6) Preparation of copolymer: Weigh HPMA (0.218g, 1.52mmol, 69mol%) into a Shleck bottle, heat and dissolve with 0.5mL DMSO, then add 0.5mL acetone; weigh the compound (0.052g, 0.12mmol, 6.5mol%) and compound (0.054g, 0.22mmol, 12mol%) was added to the Shleck bottle, stirred until dissolved, and when it was cooled to room temperature, 0.016g (5%, wt) azoisobutyronitrile (AIBN) was added, vacuumized and nitrogen cycled for 3 ~5 times, keep the temperature at about 55°C after sealing, and react for 24 hours. Precipitate with a mixture of acetone and ether (volume ratio 7:3), dissolve the precipitate with 1 mL of anhydrous methanol after filtration, and centrifuge with an ultrafiltration concentration centrifuge tube with a molecular weight of 3000 to remove small molecules and obtain 207 mg of polymer. Yield 64%.

M _n =2.7×10 ⁴ , M _w /M _n =1.28. ¹ H-NMR (400MHz,DMSO,δ,ppm):δ7.26(–CHC ₆ H ₂ H ₂ N(CH ₂ CH ₂ Cl) ₂ ), 6.79(–CHC ₆ H ₂ H ₂ N(CH ₂ CH ₂ Cl) ₂ ), 5.05, 4.14(sugarmoiety), 3.69-4.02((–OCH ₂ C(CH ₃ )( CH ₂ O–) ₂ and–O(–O)CHC ₆ H ₄ N(CH _{2 CH 2} Cl _{) 2} )),3.68(CH ₃ CH (OH)CH ₂ NH–ofHPMA),3.06(CH ₃ CH(OH)C H ₂ NH –of HPMA), 1.33–1.80 (–CH ₂ -of polymer backbone), 0.59–0.98 (–CH ₃ ).

Example 2

compound , , , , The preparation is with embodiment 1.

Preparation of copolymer: Weigh HPMA (0.159g, 1.11mmol, 82mol%) into a Shleck bottle, heat and dissolve with 0.5mL of DMSO, then add 0.5mL of acetone; weigh the compound (0.038g, 0.09mmol, 7mol%) and compound (0.023g, 0.09mmol, 7mol%) was added to the Shleck bottle, stirred until dissolved, and when it was cooled to room temperature, 0.018g (8%, wt) azoisobutyronitrile (AIBN) was added, vacuumized and filled with nitrogen for 3 ~5 times, keep the temperature at about 55°C after sealing, and react for 24 hours. Precipitate with a mixture of acetone and ether (volume ratio 7:3), filter and dissolve the precipitate with 1ml of anhydrous methanol, centrifuge with an ultrafiltration concentration centrifuge tube with a molecular weight of 3000, remove small molecules, and obtain 0.153g of polymer , yield 70%.

M _n =2.4×10 ⁴ , M _w /M _n =1.16. ¹ H-NMR (400MHz,DMSO,δ,ppm):δ7.26(–CHC ₆ H ₂ H ₂ N(CH ₂ CH ₂ Cl) ₂ ), 6.79(–CHC ₆ H ₂ H ₂ N(CH ₂ CH ₂ Cl) ₂ ), 5.05, 4.14(sugarmoiety), 3.69-4.02((–OCH ₂ C(CH ₃ )( CH ₂ O–) ₂ and–O(–O)CHC ₆ H ₄ N(CH _{2 CH 2} Cl _{) 2} )),3.68(CH ₃ CH (OH)CH ₂ NH–ofHPMA),3.06(CH ₃ CH(OH)C H ₂ NH –of HPMA), 1.33–1.80 (–CH ₂ -of polymer backbone), 0.59–0.98 (–CH ₃ ).

Example 3

compound , , , , The preparation is with embodiment 1.

Preparation of copolymer: Weigh HPMA (0.159g, 1.11mmol, 80mol%) into a Shleck bottle, heat and dissolve with 0.5mL DMSO, then add 0.5mL acetone; weigh the compound (0.057g, 0.14mmol, 10mol%) and compound 5 (0.034g, 0.14mmmol, 10mol%) were added in the Shleck bottle, stirred until dissolved, and when cooled to room temperature, 0.025g (10%, wt) of azo was added and Isobutyronitrile (AIBN), vacuumize and nitrogen cycle 3 to 5 times, keep the temperature at about 55°C after sealing, and react for 24 hours. Precipitate with a mixture of acetone and ether (volume ratio 7:3), filter and dissolve the precipitate with 1ml of anhydrous methanol, centrifuge with an ultrafiltration concentration centrifuge tube with a molecular weight of 3000, remove small molecules, and obtain 0.16g of polymer , yield 64%.

M _n =2.6×10 ⁴ , M _w /M _n =1.23. ¹ H-NMR (400MHz,DMSO,δ,ppm):δ7.26(–CHC ₆ H ₂ H ₂ N(CH ₂ CH ₂ Cl) ₂ ), 6.79(–CHC ₆ H ₂ H ₂ N(CH ₂ CH ₂ Cl) ₂ ), 5.05, 4.14(sugarmoiety), 3.69-4.02((–OCH ₂ C(CH ₃ )( CH ₂ O–) ₂ and–O(–O)CHC ₆ H ₄ N(CH _{2 CH 2} Cl _{) 2} )),3.68(CH ₃ CH (OH)CH ₂ NH–ofHPMA),3.06(CH ₃ CH(OH)C H ₂ NH –of HPMA), 1.33–1.80 (–CH ₂ -of polymer backbone), 0.59–0.98 (–CH ₃ ).

Claims (10)

1. D-galactose/benzaldehyde nitrogen mustard/N-(2-hydroxypropyl) methacrylamide copolymer, its structure is as follows: In the formula, x=80～90mol%, y=5～10mol%, z=5～10mol%; number average molecular weight Mn=2.1～2.8×10 ⁴ , Mw/Mn=1.33～1.81. 2. the preparation method of D-galactose/benzaldehyde nitrogen mustard/N-(2-hydroxypropyl) methacrylamide copolymer as claimed in claim 1, comprises the following processing steps: (1) compound Preparation: use tetrahydrofuran as solvent, p-toluenesulfonic acid as catalyst, molecular sieve as water absorbing agent, under nitrogen protection, make benzaldehyde nitrogen mustard and 1,1,1-trimethylolethane at -5～5 ℃ for 12-15 hours; after the reaction of benzaldehyde nitrogen mustard is complete, add ammonia water to neutralize p-toluenesulfonic acid to pH=7-8, filter out molecular sieves, concentrate the filtrate, dilute with dichloromethane, and use pH=8 Washed with phosphate buffer solution, dried over anhydrous sodium sulfate, and separated by silica gel column chromatography to obtain compound ; (2) compound Preparation: with dichloromethane as solvent, triethylamine as acid-binding agent, under nitrogen protection, the compound React with methacryloyl chloride at -5-0°C for 12-15 hours; after the reaction is complete, wash with saturated sodium bicarbonate, dry over anhydrous sodium sulfate, and separate by silica gel column chromatography to obtain the compound ; (3) compound Preparation of galactose: Dissolve galactose in acetone, add concentrated sulfuric acid as a dehydrating agent, stir overnight at room temperature, until the solution turns bright yellow, add potassium carbonate solution to neutralize concentrated sulfuric acid, precipitate white solid, filter with suction, and distill out acetone , extracted with chloroform, washed with distilled water, and dried over anhydrous sodium sulfate to obtain a colorless transparent oil, which is the compound ; (4) compound Preparation: Compound Dissolve in anhydrous ether, add triethylamine, cool down to -5~5°C, add methacryloyl chloride, react for 3~12 hours, the compound Dissolve in anhydrous ether, add triethylamine, cool down to -5~5°C, add methacryloyl chloride, react for 3~12 hours; remove the precipitate by suction filtration, evaporate the filtrate to dryness, and dissolve the residue with n-hexane , remove insoluble matter, distill out n-hexane to obtain a colorless viscous substance, which is the compound ; (5) compound Preparation: Compound Dissolve in trifluoroacetic acid-water mixture, stir at room temperature to fully hydrolyze, precipitate the hydrolyzate with ethanol, filter with suction to obtain a light yellow solid, recrystallize with ethanol to obtain a white solid which is the compound ; (6) Preparation of target compound: N-(2-hydroxypropyl)methacrylamide, compound and compounds Dissolve with DMSO and acetone, add initiator azobisisobutyronitrile, react at 50-60°C for 20-24 hours under nitrogen protection, precipitate with a mixture of acetone and ether, filter, dissolve the precipitate with anhydrous methanol, and finally use The ultrafiltration concentration centrifuge tube with a molecular weight of 3000 is centrifuged to remove small molecules to obtain the target high molecular weight copolymer. 3. the preparation method of D-galactose/benzaldehyde nitrogen mustard/N-(2-hydroxypropyl) methacrylamide copolymer as described in claim 2 is characterized in that: in step (1), benzene The molar ratio of formaldehyde nitrogen mustard to 1.1.1-trimethylolethane is 1:2.9～1:3.3; the molar ratio of benzaldehyde nitrogen mustard to catalyst p-toluenesulfonic acid is 1:0.05～1:0.1; The addition amount of molecular sieve is 15～20 times of the quality of benzaldehyde nitrogen mustard. 4. the preparation method of D-galactose/benzaldehyde nitrogen mustard/N-(2-hydroxypropyl) methacrylamide copolymer as claimed in claim 2, is characterized in that: in step (2), methacrylic acid The molar ratio of acid chloride to acid-binding agent triethylamine is 1:1.1～1:1; the compound The molar ratio with methacryloyl chloride is 1:1～1:1.2. 5. The preparation method of D-galactose/benzaldehyde nitrogen mustard/N-(2-hydroxypropyl)methacrylamide copolymer as claimed in claim 2, characterized in that: step (1) and step (2) Among them, the eluent used in the silica gel column chromatography is: ethyl acetate and petroleum ether in a volume ratio of 1:10 to 1:30. 6. the preparation method of D-galactose/benzaldehyde nitrogen mustard/N-(2-hydroxypropyl) methacrylamide copolymer as claimed in claim 2 is characterized in that: in step (3), the concentrated sulfuric acid The amount added is 2% to 3% of the mass of galactose; the mass concentration of potassium carbonate solution is 15% to 20%. 7. The preparation method of D-galactose/benzaldehyde nitrogen mustard/N-(2-hydroxypropyl)methacrylamide copolymer as claimed in claim 2, is characterized in that: in step (4), compound The molar ratio with methacryloyl chloride is 1:1～1:1.2; compound The molar ratio with triethylamine is 1:1.3～1:1.5; the amount of methacryloyl chloride added is the compound 0.4 to 0.6 times the mass. 8. The preparation method of D-galactose/benzaldehyde nitrogen mustard/N-(2-hydroxypropyl)methacrylamide copolymer as claimed in claim 2, is characterized in that: in step (5), trifluoroacetic acid - In the water mixture, the volume ratio of trifluoroacetic acid to water is 9:1-8:1. 9. The preparation method of D-galactose/benzaldehyde nitrogen mustard/N-(2-hydroxypropyl)methacrylamide copolymer as claimed in claim 2, is characterized in that: in step (6), the compound , compound , N-(2-hydroxypropyl) methacrylamide molar ratio is 1:1:4～1:1:20; The consumption of described initiator azobisisobutyronitrile is compound , compound and 5%-10% of the total mass of N-(2-hydroxypropyl)methacrylamide; in the mixture of acetone and ether, the volume ratio of acetone and ether is 7:3-7:1. 10. D-galactose/benzaldehyde nitrogen mustard/N-(2-hydroxypropyl) methacrylamide copolymer as claimed in claim 1 is used in the preparation of anticancer drugs as liver cancer HepG2 cell inhibitor.