Anticancer drug adjuvant with sustained-release function and preparation method thereof

Abstract

The present invention discloses a method for preparing an anticancer drug adjuvant with sustained-release function, comprising the following steps: S1, preparation of mesoporous alumina; S2, preparation of pretreated mesoporous alumina; S3, preparation of organic mesoporous alumina; S4, preparation of modified mesoporous alumina; S5, preparation of anticancer drug adjuvant. The anticancer drug adjuvant with sustained-release function provided by the present invention is prepared by multi-step surface modification to obtain a multifunctional anticancer drug adjuvant, which has excellent mesoporous structure, cyclodextrin inclusion complex, folic acid targeting and drug absorption promotion, etc. It can not only achieve efficient drug loading and controllable release, but also has the ability of active targeting and drug absorption promotion, thereby improving the therapeutic effect of anticancer drugs and improving the bioavailability of anticancer drugs.

Description

Technical Field

[0001] The invention belongs to the technical field of drug carriers, and in particular relates to an anticancer drug adjuvant with a sustained-release function and a preparation method thereof. Background Art

[0002] Drug sustained-release materials, also known as drug sustained-release carrier materials, are carriers of small molecule drugs in the slow release process of drugs and are an important part of drug sustained-release systems. The speed and efficacy of drug release are generally regulated by controlling the structural changes of drug carrier materials. At present, drug sustained-release carrier materials generally use polymer carrier materials, which are commonly used natural polymer carrier materials and synthetic polymer carrier materials.

[0003] Developing effective drug carriers, reducing the intrinsic adverse reactions of chemotherapeutic drugs, and improving the therapeutic effect have become one of the key issues in the treatment of cancer. At present, drug nanocarriers have been developed to improve the accumulation of drugs in tumor areas in order to enhance the permeability and retention effect, and the nanocarriers include polymer vesicles, micelles, polymer nanoparticles, inorganic nanoparticles, and hybrid porous solids.

[0004] Chinese patent application CN108078939A discloses an anticancer drug carrier preparation and its application. The present invention uses succinic anhydride, Lycium barbarum polysaccharide and α-linolenic acid as reaction raw materials, first reacts succinic anhydride and Lycium barbarum polysaccharide under alkaline conditions, puts the reactants into dimethyl sulfoxide solvent, activates them under the action of N'N-carbonyldiimidazole, and further esterifies the activated substances with α-linolenic acid to obtain a target product with high mechanical strength, good stability and excellent biocompatibility. The prepared drug carrier is used for loading hydrophobic tumor drugs to improve the bioavailability of tumor drugs. However, the loading capacity and sustained-release stability of the carrier need to be further improved. Summary of the invention

[0005] In view of the deficiencies of the prior art, the purpose of the present invention is to provide an anticancer drug adjuvant with sustained-release function and a preparation method thereof, wherein the anticancer drug adjuvant has a large drug loading capacity and releases the drug smoothly, thereby improving the bioavailability of the drug.

[0006] To achieve the above object, the present invention provides the following technical solutions:

[0007] A method for preparing an anticancer drug adjuvant with sustained-release function comprises the following steps:

[0008] S1. Preparation of mesoporous alumina;

[0009] S2. Preparation of pretreated mesoporous alumina: adding mesoporous alumina to an ethanol aqueous solution, and then adding γ-glycidyloxypropyltrimethoxysilane, stirring to react, filtering, washing, and drying after the reaction is completed to obtain pretreated mesoporous alumina;

[0010] S3, preparation of organic mesoporous alumina: adding the pretreated mesoporous alumina in step S2 into DMF, then adding hydroxypropyl cyclodextrin and triethylamine, heating and reacting under nitrogen protection, filtering, washing and drying after the reaction is completed to obtain organic mesoporous alumina;

[0011] S4, preparation of modified mesoporous alumina: adding 3-mercaptopropionic acid and folic acid to a DMF solution, then adding EDC and NHS, stirring and activating, adding the organic mesoporous alumina in step S3 after activation, carrying out esterification reaction under nitrogen protection, filtering, washing and drying after the reaction to obtain modified mesoporous alumina;

[0012] S5. Preparation of anticancer drug adjuvant: adding the modified mesoporous alumina in step S4 into DMF, and then adding 8-shogaol and azobisisobutyronitrile, and reacting at a constant temperature. After the reaction is completed, filtering, washing and drying are performed to obtain the anticancer drug adjuvant.

[0013] Preferably, the preparation method of the mesoporous alumina in step S1 is as follows:

[0014] Hexadecyltrimethylammonium bromide is added into deionized water, stirred to dissolve, and then aluminum nitrate is added. After stirring evenly, urea is added to carry out a hydrothermal reaction. After the reaction is completed, aging is carried out, and then filtering, washing, drying and calcining are carried out to obtain the mesoporous alumina.

[0015] Preferably, the mass ratio of hexadecyltrimethylammonium bromide, deionized water, aluminum nitrate and urea is 6-8:300-400:25-30:12-16, the temperature of the hydrothermal reaction is 120-140°C, and the time is 6-9h; the temperature of the aging is 70-80°C, and the time is 10-15h; the temperature of the calcination is 500-600°C, and the time is 3-4h.

[0016] In the present invention, mesoporous alumina is prepared by hydrothermal method. Mesoporous alumina has the characteristics of high specific surface area, large pore volume, adjustable pore size distribution, suitable surface acidity and good thermal stability, and can effectively increase the loading amount of the auxiliary agent for the drug.

[0017] Preferably, the mass concentration of ethanol in the ethanol aqueous solution in step S2 is 70-80%, the mass ratio of the mesoporous alumina to γ-glycidyloxypropyltrimethoxysilane is 50:10-14, the temperature of the stirring reaction is 70-75°C, and the time is 4-6h.

[0018] In the present invention, γ-glycidyloxypropyltrimethoxysilane is used to modify the surface of mesoporous alumina, thereby introducing epoxy groups on the surface of alumina, which is beneficial to the subsequent reaction.

[0019] Preferably, in step S3, the mass ratio of the pretreated mesoporous alumina, hydroxypropyl cyclodextrin and triethylamine is 50:12-18:0.5-1, the temperature of the heating reaction is 60-70° C., and the time is 3-5 h.

[0020] In the present invention, hydroxypropyl cyclodextrin is introduced onto alumina by means of covalent bonds. Cyclodextrin has a hydrophobic inner cavity, which can include drug molecules to improve the solubility of the drug, and can also have the function of sustained release of the drug, so that the drug can be released smoothly.

[0021] Preferably, the mass ratio of 3-mercaptopropionic acid, folic acid, EDC, NHS and organic mesoporous alumina in step S4 is 10-13:5-7:10-15:13-18:50-60.

[0022] Preferably, the stirring activation temperature in step S4 is 20-30°C, the rotation speed is 150-200r / min, and the time is 1-2h; the esterification reaction temperature is 40-50°C, and the time is 3-4h.

[0023] In the present invention, 3-mercaptopropionic acid and folic acid are introduced onto an alumina carrier by an esterification reaction between the carboxyl groups in 3-mercaptopropionic acid and folic acid and the hydroxyl groups in hydroxypropyl cyclodextrin. Folic acid can bind to the folic acid receptors overexpressed on the surface of cancer cells, giving the adjuvant targeting properties. The thiol group in 3-mercaptopropionic acid is beneficial to the subsequent reactions.

[0024] Preferably, the mass ratio of the modified mesoporous alumina, 8-shogaol and azobisisobutyronitrile in step S5 is 50-60:4-7:0.5-1.

[0025] Preferably, the isothermal reaction in step S5 is carried out at a temperature of 60-70° C. and for a time of 1-2 h.

[0026] In the present invention, 8-gingerol is introduced onto an alumina carrier through a thiol-ene reaction. 8-gingerol has strong antioxidant and anti-inflammatory effects, and can improve the stability of anticancer drugs. At the same time, 8-gingerol has moderate fat solubility, can interact with the phospholipid bilayer of the cell membrane, and can temporarily change the fluidity and permeability of the cell membrane, thereby promoting the absorption of drug molecules, improving the bioavailability of the drug, and enhancing the drug delivery effect of the auxiliary agent.

[0027] The present invention also protects an anticancer drug adjuvant with sustained-release function prepared by the method as described above.

[0028] Compared with the prior art, the present invention has the following beneficial effects:

[0029] (1) The anticancer drug adjuvant with sustained release function provided by the present invention is prepared through multi-step surface modification to obtain a multifunctional anticancer drug adjuvant. The adjuvant has excellent mesoporous structure, cyclodextrin inclusion complex, folic acid targeting and drug absorption promotion effects. It can not only achieve efficient drug loading and controllable release, but also has the ability to actively target and promote drug absorption, thereby improving the therapeutic effect of anticancer drugs and improving the bioavailability of anticancer drugs.

[0030] (2) The anticancer drug adjuvant with sustained-release function provided by the present invention adopts hexadecyltrimethylammonium bromide as a template and prepares mesoporous alumina by hydrothermal reaction. The obtained mesoporous alumina carrier has a regular mesoporous structure and a large specific surface area, which provides a good foundation for subsequent functional modification and drug loading. Subsequently, epoxy groups are introduced into the surface of the mesoporous alumina, which not only enhances the chemical stability of the alumina carrier, but also provides sufficient reaction sites for subsequent functional modification.

[0031] (3) The anticancer drug adjuvant with sustained release function provided by the present invention is characterized in that the organic mesoporous alumina is reacted with hydroxypropyl cyclodextrin, and the hydrophobic inner cavity of the cyclodextrin molecule can form an inclusion complex with the drug molecule. This host-guest interaction realizes the controlled sustained release of the drug and effectively prolongs the in vivo circulation time of the drug. Subsequently, the modified mesoporous alumina is reacted with 3-mercaptopropionic acid and folic acid, and the folic acid molecules are grafted onto the carrier in the form of covalent bonds, so that the carrier has the ability to recognize the folic acid receptors overexpressed on the surface of cancer cells, thereby realizing active targeting of tumor tissues and improving the enrichment efficiency of the drug in the lesion site. At the same time, the thiol group introduced into the 3-mercaptopropionic acid is conducive to the subsequent reaction. Finally, the modified mesoporous alumina is reacted with 8-gingerol. On the one hand, 8-gingerol can effectively remove free radicals generated during drug delivery, protect drug molecules from oxidative damage, and improve the stability of the drug delivery system. On the other hand, it can enhance cell permeability and improve the absorption rate of the drug at the target site, thereby improving the cell uptake efficiency, bioavailability and therapeutic effect of the drug. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] Figure 1 This is a graph of the cumulative release rates of different groups of anticancer drug adjuvants loaded with anticancer drugs in a simulated small intestinal environment. DETAILED DESCRIPTION

[0033] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0034] Example 1

[0035] A method for preparing an anticancer drug adjuvant with sustained-release function comprises the following steps:

[0036] S1. Preparation of mesoporous alumina: 7 g of hexadecyltrimethylammonium bromide was added to 350 g of deionized water, stirred to dissolve, then 28 g of aluminum nitrate was added, stirred evenly, 14 g of urea was added, and the mixture was hydrothermally reacted at 130° C. for 8 h. After the reaction was completed, the mixture was aged at 75° C. for 13 h, then filtered, washed, dried, and calcined at 550° C. for 3.5 h to obtain mesoporous alumina.

[0037] S2. Preparation of pretreated mesoporous alumina: 50 g of mesoporous alumina was added to 800 mL of ethanol aqueous solution (the mass concentration of ethanol was 75%), followed by adding 12 g of γ-glycidyloxypropyltrimethoxysilane, and stirred at 72° C. for 5 h. After the reaction was completed, the mixture was filtered, washed, and dried to obtain pretreated mesoporous alumina.

[0038] S3, preparation of organic mesoporous alumina: add 50g of the pretreated mesoporous alumina in step S2 into 700mL of DMF, then add 15g of hydroxypropyl cyclodextrin and 0.8g of triethylamine, react at 65°C for 4h under nitrogen protection, filter, wash and dry after the reaction to obtain organic mesoporous alumina;

[0039] S4, preparation of modified mesoporous alumina: 12g 3-mercaptopropionic acid and 6g folic acid were added to 800mL DMF solution, followed by 13g EDC and 16g NHS, and the mixture was stirred and activated at a speed of 180r / min and a temperature of 25°C for 1.5h. After activation, 55g of the organic mesoporous alumina in step S3 was added, and the mixture was reacted at 45°C for 3.5h under nitrogen protection. After the reaction was completed, the mixture was filtered, washed, and dried to obtain modified mesoporous alumina;

[0040] S5. Preparation of anticancer drug adjuvant: 55 g of modified mesoporous alumina in step S4 was added into 800 mL of DMF, followed by 6 g of 8-shogaol and 0.8 g of azobisisobutyronitrile, and the mixture was reacted at 65° C. for 1.5 h. After the reaction was completed, the mixture was filtered, washed and dried to obtain the anticancer drug adjuvant.

[0041] Example 2

[0042] A method for preparing an anticancer drug adjuvant with sustained-release function comprises the following steps:

[0043] S1. Preparation of mesoporous alumina: 7 g of hexadecyltrimethylammonium bromide was added to 350 g of deionized water, stirred to dissolve, then 27 g of aluminum nitrate was added, stirred evenly, 14 g of urea was added, and the mixture was hydrothermally reacted at 130° C. for 7 h. After the reaction was completed, the mixture was aged at 75° C. for 12 h, then filtered, washed, dried, and calcined at 550° C. for 3.5 h to obtain mesoporous alumina.

[0044] S2. Preparation of pretreated mesoporous alumina: 50 g of mesoporous alumina was added to 800 mL of ethanol aqueous solution (the mass concentration of ethanol was 75%), followed by adding 12 g of γ-glycidyloxypropyltrimethoxysilane, and stirred at 72° C. for 5 h. After the reaction was completed, the mixture was filtered, washed, and dried to obtain pretreated mesoporous alumina.

[0045] S3, preparation of organic mesoporous alumina: add 50g of the pretreated mesoporous alumina in step S2 into 700mL of DMF, then add 14g of hydroxypropyl cyclodextrin and 0.7g of triethylamine, react at 65°C for 4h under nitrogen protection, filter, wash and dry after the reaction to obtain organic mesoporous alumina;

[0046] S4, preparation of modified mesoporous alumina: 11g 3-mercaptopropionic acid and 6g folic acid were added to 800mL DMF solution, followed by 12g EDC and 15g NHS, and the mixture was stirred and activated at a speed of 170r / min and a temperature of 25°C for 1.5h. After the activation, 55g of the organic mesoporous alumina in step S3 was added, and the mixture was reacted at 45°C for 3.5h under nitrogen protection. After the reaction was completed, the mixture was filtered, washed, and dried to obtain modified mesoporous alumina;

[0047] S5. Preparation of anticancer drug adjuvant: 55 g of modified mesoporous alumina in step S4 was added into 800 mL of DMF, followed by 5 g of 8-shogaol and 0.7 g of azobisisobutyronitrile, and the mixture was reacted at 65° C. for 1.5 h. After the reaction was completed, the mixture was filtered, washed and dried to obtain the anticancer drug adjuvant.

[0048] Example 3

[0049] A method for preparing an anticancer drug adjuvant with sustained-release function comprises the following steps:

[0050] S1. Preparation of mesoporous alumina: 6 g of hexadecyltrimethylammonium bromide was added to 300 g of deionized water, stirred to dissolve, then 25 g of aluminum nitrate was added, stirred evenly, 12 g of urea was added, and the mixture was hydrothermally reacted at 120° C. for 9 h. After the reaction was completed, the mixture was aged at 70° C. for 15 h, then filtered, washed, dried, and calcined at 500° C. for 4 h to obtain mesoporous alumina.

[0051] S2. Preparation of pretreated mesoporous alumina: 50 g of mesoporous alumina was added to 800 mL of ethanol aqueous solution (the mass concentration of ethanol was 70%), followed by adding 10 g of γ-glycidyloxypropyltrimethoxysilane, and stirred at 70° C. for 6 h. After the reaction was completed, the mixture was filtered, washed, and dried to obtain pretreated mesoporous alumina.

[0052] S3, preparation of organic mesoporous alumina: add 50g of the pretreated mesoporous alumina in step S2 into 700mL of DMF, then add 12g of hydroxypropyl cyclodextrin and 0.5g of triethylamine, react at 60°C for 5h under nitrogen protection, filter, wash and dry after the reaction to obtain organic mesoporous alumina;

[0053] S4, preparation of modified mesoporous alumina: add 10g 3-mercaptopropionic acid and 5g folic acid to 800mL DMF solution, then add 10g EDC and 13g NHS, stir and activate at a speed of 150r / min and a temperature of 20°C for 2h, add 50g organic mesoporous alumina in step S3 after activation, react at 40°C for 4h under nitrogen protection, filter, wash and dry after the reaction to obtain modified mesoporous alumina;

[0054] S5. Preparation of anticancer drug adjuvant: 50 g of modified mesoporous alumina in step S4 was added into 800 mL of DMF, followed by 4 g of 8-shogaol and 0.5 g of azobisisobutyronitrile, and the mixture was reacted at 60° C. for 2 h. After the reaction was completed, the mixture was filtered, washed and dried to obtain the anticancer drug adjuvant.

[0055] Example 4

[0056] A method for preparing an anticancer drug adjuvant with sustained-release function comprises the following steps:

[0057] S1. Preparation of mesoporous alumina: 8 g of hexadecyltrimethylammonium bromide was added to 400 g of deionized water, stirred to dissolve, then 30 g of aluminum nitrate was added, stirred evenly, 16 g of urea was added, and the mixture was hydrothermally reacted at 140° C. for 6 h. After the reaction was completed, the mixture was aged at 80° C. for 10 h, then filtered, washed, dried, and calcined at 600° C. for 3 h to obtain mesoporous alumina.

[0058] S2. Preparation of pretreated mesoporous alumina: 50 g of mesoporous alumina was added to 800 mL of ethanol aqueous solution (the mass concentration of ethanol was 80%), followed by adding 14 g of γ-glycidyloxypropyltrimethoxysilane, and stirred at 75° C. for 4 h. After the reaction was completed, the mixture was filtered, washed, and dried to obtain pretreated mesoporous alumina.

[0059] S3, preparation of organic mesoporous alumina: add 50g of the pretreated mesoporous alumina in step S2 into 700mL of DMF, then add 18g of hydroxypropyl cyclodextrin and 1g of triethylamine, react at 70°C for 3h under nitrogen protection, filter, wash and dry after the reaction to obtain organic mesoporous alumina;

[0060] S4, preparation of modified mesoporous alumina: 13g 3-mercaptopropionic acid and 7g folic acid were added to 800mL DMF solution, followed by 15g EDC and 18g NHS, and the mixture was stirred and activated at a speed of 200r / min and a temperature of 30°C for 1h. After activation, 60g of the organic mesoporous alumina in step S3 was added, and the mixture was reacted at 50°C for 3h under nitrogen protection. After the reaction was completed, the mixture was filtered, washed, and dried to obtain modified mesoporous alumina;

[0061] S5. Preparation of anticancer drug adjuvant: add 60g modified mesoporous alumina in step S4 into 800mL DMF, then add 7g 8-shogaol and 1g azobisisobutyronitrile, and react at 70°C for 1h. After the reaction is completed, filter, wash and dry to obtain the anticancer drug adjuvant.

[0062] Comparative Example 1

[0063] A method for preparing an anticancer drug adjuvant with sustained-release function comprises the following steps:

[0064] S1. Preparation of mesoporous alumina: 7 g of hexadecyltrimethylammonium bromide was added to 350 g of deionized water, stirred to dissolve, then 28 g of aluminum nitrate was added, stirred evenly, 14 g of urea was added, and the mixture was hydrothermally reacted at 130° C. for 8 h. After the reaction was completed, the mixture was aged at 75° C. for 13 h, then filtered, washed, dried, and calcined at 550° C. for 3.5 h to obtain mesoporous alumina.

[0065] S2. Preparation of pretreated mesoporous alumina: 50 g of mesoporous alumina was added to 800 mL of ethanol aqueous solution (the mass concentration of ethanol was 75%), followed by adding 12 g of γ-glycidyloxypropyltrimethoxysilane, and stirred at 72° C. for 5 h. After the reaction was completed, the mixture was filtered, washed, and dried to obtain pretreated mesoporous alumina.

[0066] S3, preparation of modified mesoporous alumina: 12g 3-mercaptopropionic acid and 6g folic acid were added to 800mL DMF solution, followed by 13g EDC and 16g NHS, and the mixture was stirred and activated at a speed of 180r / min and a temperature of 25°C for 1.5h. After activation, 55g pretreated mesoporous alumina in step S2 was added, and the mixture was reacted at 45°C for 3.5h under nitrogen protection. After the reaction was completed, the mixture was filtered, washed, and dried to obtain modified mesoporous alumina;

[0067] S4, preparation of anticancer drug adjuvant: add 55g modified mesoporous alumina in step S3 into 800mL DMF, then add 6g 8-shogaol and 0.8g azobisisobutyronitrile, and react at 65°C for 1.5h. After the reaction is completed, filter, wash and dry to obtain the anticancer drug adjuvant.

[0068] Compared with Example 1, hydroxypropyl cyclodextrin was not introduced into this comparative example.

[0069] Comparative Example 2

[0070] A method for preparing an anticancer drug adjuvant with sustained-release function comprises the following steps:

[0071] S1. Preparation of mesoporous alumina: 7 g of hexadecyltrimethylammonium bromide was added to 350 g of deionized water, stirred to dissolve, then 28 g of aluminum nitrate was added, stirred evenly, 14 g of urea was added, and the mixture was hydrothermally reacted at 130° C. for 8 h. After the reaction was completed, the mixture was aged at 75° C. for 13 h, then filtered, washed, dried, and calcined at 550° C. for 3.5 h to obtain mesoporous alumina.

[0072] S2. Preparation of pretreated mesoporous alumina: 50 g of mesoporous alumina was added to 800 mL of ethanol aqueous solution (the mass concentration of ethanol was 75%), followed by adding 12 g of γ-glycidyloxypropyltrimethoxysilane, and stirred at 72° C. for 5 h. After the reaction was completed, the mixture was filtered, washed, and dried to obtain pretreated mesoporous alumina.

[0073] S3, preparation of organic mesoporous alumina: add 50g of the pretreated mesoporous alumina in step S2 into 700mL of DMF, then add 15g of hydroxypropyl cyclodextrin and 0.8g of triethylamine, react at 65°C for 4h under nitrogen protection, filter, wash and dry after the reaction to obtain organic mesoporous alumina;

[0074] S4, preparation of modified mesoporous alumina: add 6g of folic acid to 800mL of DMF solution, then add 13g of EDC and 16g of NHS, stir and activate at a speed of 180r / min and a temperature of 25°C for 1.5h, add 55g of the organic mesoporous alumina in step S3 after activation, react at 45°C for 3.5h under nitrogen protection, filter, wash and dry after the reaction to obtain modified mesoporous alumina;

[0075] S5. Preparation of anticancer drug adjuvant: 55 g of modified mesoporous alumina in step S4 was added into 800 mL of DMF, followed by 6 g of 8-shogaol, and the mixture was oscillated at a constant temperature of 65° C. for 1.5 h. After the reaction was completed, the mixture was filtered, washed, and dried to obtain the anticancer drug adjuvant.

[0076] Compared with Example 1, in step S4 of this comparative example, no 3-mercaptopropionic acid is added, and in step S5, 8-shogaol is introduced onto the mesoporous alumina by adsorption.

[0077] Comparative Example 3

[0078] A method for preparing an anticancer drug adjuvant with sustained-release function comprises the following steps:

[0079] S1. Preparation of mesoporous alumina: 7 g of hexadecyltrimethylammonium bromide was added to 350 g of deionized water, stirred to dissolve, then 28 g of aluminum nitrate was added, stirred evenly, 14 g of urea was added, and the mixture was hydrothermally reacted at 130° C. for 8 h. After the reaction was completed, the mixture was aged at 75° C. for 13 h, then filtered, washed, dried, and calcined at 550° C. for 3.5 h to obtain mesoporous alumina.

[0080] S2. Preparation of pretreated mesoporous alumina: 50 g of mesoporous alumina was added to 800 mL of ethanol aqueous solution (the mass concentration of ethanol was 75%), followed by adding 12 g of γ-glycidyloxypropyltrimethoxysilane, and stirred at 72° C. for 5 h. After the reaction was completed, the mixture was filtered, washed, and dried to obtain pretreated mesoporous alumina.

[0081] S3, preparation of organic mesoporous alumina: add 50g of the pretreated mesoporous alumina in step S2 into 700mL of DMF, then add 15g of hydroxypropyl cyclodextrin and 0.8g of triethylamine, react at 65°C for 4h under nitrogen protection, filter, wash and dry after the reaction to obtain organic mesoporous alumina;

[0082] S4, preparation of modified mesoporous alumina: add 12g 3-mercaptopropionic acid and 6g folic acid into 800mL DMF solution, then add 13g EDC and 16g NHS, stir and activate at a speed of 180r / min and a temperature of 25°C for 1.5h, add 55g of the organic mesoporous alumina in step S3 after the activation, react at 45°C for 3.5h under nitrogen protection, filter, wash and dry after the reaction to obtain the anticancer drug adjuvant.

[0083] Compared with Example 1, this comparative example does not introduce 8-shogaol.

[0084] The anticancer drug adjuvants prepared in Examples 1-4 and Comparative Examples 1-3 were subjected to drug loading tests, as follows:

[0085] 1. Drug loading test

[0086] The anticancer drug adjuvant (100 mg) prepared in Examples 1-4 and Comparative Examples 1-3 were added to an anticancer drug solution (20 mL, 5 mg / mL, ethanol solution of paclitaxel), adsorbed at 60°C and 160 rpm for 8 h, and the supernatant was centrifuged. The drug concentration in the supernatant was tested by ultraviolet spectrophotometry, and the drug loading of the anticancer drug adjuvant to the drug was indirectly determined. Drug loading rate (%) = 100%, where m0 is the mass of the anticancer drug (mg), m1 is the mass of the anticancer drug in the supernatant (mg), and m2 is the mass of the anticancer drug adjuvant (mg). The test results are shown in Table 1.

[0087] 2. In vitro drug release study

[0088] The drug-loaded anticancer drug adjuvants prepared in Example 1 and Comparative Examples 1-2 were subjected to in vitro release experiments at a physiological temperature of 37°C using a dialysis method. The small intestine environment was simulated with a phosphate buffer solution (pH=6.8). 100 mg of the drug-loaded sample was placed in a dialysis bag (1000Da), and the dialysis bag was immersed in a beaker containing 60.0 mL of PBS buffer, and stirred at a constant speed of 100 rpm using a magnetic bar. At regular intervals, 3.0 mL of the release medium was taken out, and the same volume of fresh release medium was added to the beaker. The absorbance of the drug was measured by an ultraviolet spectrophotometer, and the cumulative release rate of the drug was calculated. The results are as follows. Figure 1 .

[0089] 3. Antioxidant research

[0090] Using the DPPH free radical scavenging activity method, the anticancer drug adjuvants prepared in Examples 1-4 and Comparative Examples 1-3 were dispersed into dispersions with a concentration of 1 mg / mL using ethanol, and the concentration of the DPPH ethanol solution was prepared to 6×10-5 mol / L. Add 2.0mL of sample dispersion to 1.0mL of DPPH ethanol solution, mix well with a vortex mixer, and react for 30 minutes in the dark. Use an ultraviolet spectrophotometer to measure the absorbance in the wavelength range of 450-650mm, determine the measurement wavelength, measure the absorbance of the sample at this wavelength, and record it as A1. An equal amount of DPPH ethanol solution is used as a blank control group, and its absorbance is recorded as A0. The results are shown in Table 1 below.

[0091] DPPH clearance rate = (1-A1 / A0)×100%

[0092] 4. The drug-carrying anticancer drug adjuvant prepared in Example 1 and Comparative Examples 1-3 was subjected to an experiment to inhibit tumor cell growth in mice, as follows:

[0093] The HELA cell line was tested by MTT reduction method. HELA cells were selected for routine culture. When the cells were expanded to a certain number and in good growth state, HELA cells in logarithmic growth phase were digested with 0.25% trypsin for 2 min, centrifuged at 1000 rpm for 5 min, the supernatant was discarded, and the cell pellet was resuspended in fresh serum-free DMEM medium to prepare 1×10 6 Then, the cell suspension was aspirated with a syringe, the air bubbles were evacuated, and the suspension was injected into the subcutaneous tissue of the left armpit of BALB / C female mice weighing about 20-22g. Each mouse was inoculated with 100 μL, containing about 1×10 5 The cells were inoculated into 50 mice in total. The tumor growth of the mice was observed, and the mice were weighed regularly to measure the tumor size. During the experiment, the animals had normal diet and water.

[0094] When the tumor grows to 100-150 mm 3 The mice were randomly divided into 5 groups, each group had 10 nude mice, namely negative control group (normal saline group), Example 1 group, Comparative Example 1 group, Comparative Example 2 group, Comparative Example 3 group, and the drugs were administered once every other day. After the administration, the mice were raised normally, the survival status of the mice was observed every day, the weight of the mice was recorded every two days, and the long and short diameters of the mouse tumors were measured with a vernier caliper to calculate the tumor volume (V).

[0095] Tumor inhibition rate = (1-V1 / V2) × 100%, where V1 is the tumor volume of the drug-treated group on the 12th day, and V2 is the tumor volume of the saline group on the 12th day. The results are shown in Table 2 below.

[0096] Table 1 Drug loading and antioxidant activity of different groups of anticancer drug adjuvants

[0097]

[0098] Table 2 Tumor inhibition rate of different groups of anticancer drug adjuvants

[0099]

[0100] It can be seen from Table 1 above that the anticancer drug adjuvant prepared by the present invention has a large drug loading capacity and a high DPPH clearance rate, which can improve the stability and biological activity of anticancer drugs; it can be seen from Table 2 that the drug-loaded anticancer drug adjuvant prepared by the present invention has a high tumor inhibition rate on mouse tumors, indicating that the anticancer drug adjuvant of the present invention can assist anticancer drugs to play a role and improve the bioavailability of anticancer drugs; Figure 1 It can be seen that the anticancer drug prepared by the present invention has a stable drug release rate and has good application prospects.

[0101] Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and variations may be made to the embodiments without departing from the principles and spirit of the present invention, and that the scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. A method for preparing an anticancer drug adjuvant with sustained-release function, characterized in that: The following steps are involved: S1. Preparation of mesoporous alumina: adding hexadecyltrimethylammonium bromide to deionized water, stirring to dissolve, then adding aluminum nitrate, stirring evenly, adding urea, and performing a hydrothermal reaction. After the reaction is completed, aging is performed, and then filtering, washing, drying, and calcining are performed to obtain the mesoporous alumina; S2, adding mesoporous alumina to an ethanol aqueous solution, and then adding γ-glycidyloxypropyltrimethoxysilane, stirring to react, filtering, washing, and drying after the reaction is completed to obtain pretreated mesoporous alumina; S3, adding the pretreated mesoporous alumina in step S2 into DMF, then adding hydroxypropyl cyclodextrin and triethylamine, heating and reacting under nitrogen protection, filtering, washing and drying after the reaction is completed to obtain organic mesoporous alumina; S4, adding 3-mercaptopropionic acid and folic acid to the DMF solution, and then adding EDC and NHS, stirring and activating, adding the organic mesoporous alumina in step S3 after the activation, and performing esterification reaction under nitrogen protection, filtering, washing and drying after the reaction to obtain modified mesoporous alumina; S5, adding the modified mesoporous alumina in step S4 into DMF, and then adding 8-shogaol and azobisisobutyronitrile, and reacting at a constant temperature. After the reaction is completed, filtering, washing, and drying are performed to obtain the anticancer drug adjuvant.

2. The preparation method according to claim 1, characterized in that: The mass ratio of hexadecyltrimethylammonium bromide, deionized water, aluminum nitrate and urea is 6-8:300-400:25-30:12-16, the temperature of the hydrothermal reaction is 120-140°C, and the time is 6-9h; the temperature of the aging is 70-80°C, and the time is 10-15h; the temperature of the calcination is 500-600°C, and the time is 3-4h.

3. The preparation method according to claim 1, characterized in that: The mass concentration of ethanol in the ethanol aqueous solution in step S2 is 70-80%, the mass ratio of the mesoporous alumina to γ-glycidyloxypropyltrimethoxysilane is 50:10-14, the temperature of the stirring reaction is 70-75°C, and the time is 4-6h.

4. The preparation method according to claim 1, characterized in that: In step S3, the mass ratio of the pretreated mesoporous alumina, hydroxypropyl cyclodextrin and triethylamine is 50:12-18:0.5-1, the temperature of the heating reaction is 60-70° C., and the time is 3-5 hours.

5. The preparation method according to claim 1, characterized in that: The mass ratio of 3-mercaptopropionic acid, folic acid, EDC, NHS and organic mesoporous alumina in step S4 is 10-13:5-7:10-15:13-18:50-60.

6. The preparation method according to claim 1, characterized in that: The stirring activation in step S4 is carried out at a temperature of 20-30°C, a rotation speed of 150-200 r / min, and a time of 1-2 h; the esterification reaction is carried out at a temperature of 40-50°C and a time of 3-4 h.

7. The preparation method according to claim 1, characterized in that: The mass ratio of the modified mesoporous alumina, 8-shogaol and azobisisobutyronitrile in step S5 is 50-60:4-7:0.5-1.

8. The preparation method according to claim 1, characterized in that: The isothermal reaction in step S5 is carried out at a temperature of 60-70° C. and for a time of 1-2 h.

9. An anticancer drug adjuvant with sustained release function prepared by the method according to any one of claims 1 to 8.

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