Active oxygen responsive paclitaxel prodrug, high-drug-loading nano preparation thereof and preparation method of high-drug-loading nano preparation

Abstract

The invention relates to the technical field of medicines, and particularly provides an active oxygen responsive paclitaxel prodrug which has a structure as shown in a formula I or a formula II. Aiming at the problem of hydrophobic aggregation of paclitaxel, the paclitaxel drug is modified, the water solubility of the modified paclitaxel drug is greatly enhanced, the self-assembly performance is obtained, the modified paclitaxel drug can be self-assembled in an aqueous solution to obtain the nano particles, and the nano particles have ultrahigh drug loading capacity and drug loading efficiency. The paclitaxel nano-preparation prepared by the method can be used for preparing the paclitaxel nano-preparation, can specifically respond to high-concentration active oxygen in a tumor microenvironment, not only can improve the insufficiency of drug loading capacity of a traditional paclitaxel nano-preparation, but also can respond to release paclitaxel at a tumor position, is beneficial to reduction of drug dosage and reduction of toxic and side effects, and has important significance on improvement of patient prognosis and prolonging of patient lifetime.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical technology, and in particular to an active oxygen-responsive paclitaxel prodrug and a high-drug-loaded nano preparation thereof and a preparation method thereof. Background Art

[0002] Paclitaxel (PTX) is a taxane diterpenoid compound isolated from the Pacific yew tree. As a plant molecule, paclitaxel has become one of the star anticancer drugs due to its excellent therapeutic effect. However, side effects such as poor water solubility and systemic toxicity limit its further clinical application. In order to avoid these shortcomings, a series of drug delivery systems have been developed in clinical practice, such as GenexolPM, Nanoxel, Abraxane and Apealea. However, the strong hydrophobicity of paclitaxel still leads to a low drug loading, and the drug loading of commercial paclitaxel preparations generally does not exceed 20%.

[0003] Currently, paclitaxel nanoparticles used in clinical practice, such as Taxol, Abraxane, and Apealea, are mostly loaded with paclitaxel using various biological or polymer materials as drug carriers. Due to the strong hydrophobicity of paclitaxel itself, its nanoparticles usually show low drug loading and efficiency (generally <

[0004] 10%); some nano drugs with drug loading greater than 10% are achieved by improving the preparation process, but this also leads to an increase in the complexity of the drug formulation; and the paclitaxel nano preparations currently used in clinical practice have basically no responsive release ability and have strong systemic toxicity and side effects. Summary of the invention

[0005] In view of this, the technical problem to be solved by the present invention is to provide an active oxygen-responsive paclitaxel prodrug and a high-drug loading nanoformulation and a preparation method thereof. The active oxygen-responsive paclitaxel prodrug provided by the present invention has a high drug loading amount and drug loading efficiency, and can release paclitaxel in response to the tumor location.

[0006] To achieve the above object, the present invention provides an active oxygen-responsive paclitaxel prodrug having a structure shown in Formula I or Formula II:

[0007]

[0008] The paclitaxel prodrug provided by the present invention can respond to over-expressed reactive oxygen species in the tumor microenvironment. The sensitive groups in the prodrug will be cleaved under reactive oxygen conditions and release paclitaxel. Therefore, the prodrug has the ability to respond to and release reactive oxygen species, and systemic toxicity and side effects are reduced.

[0009] The present invention provides a method for preparing the active oxygen-responsive paclitaxel prodrug shown in the above formula I, comprising the following steps:

[0010] The paclitaxel is reacted with the N,N'-carbonyldiimidazole-modified 4-(hydroxymethyl)phenylboronic acid pinacol ester shown in the structure of formula III to obtain the active oxygen-responsive paclitaxel prodrug shown in formula I;

[0011]

[0012] The N,N'-carbonyldiimidazole-modified 4-(hydroxymethyl)phenylboronic acid pinacol ester shown in the structure of formula III is obtained by reacting 4-(hydroxymethyl)phenylboronic acid pinacol ester and N,N'-carbonyldiimidazole.

[0013] The reaction equation is as follows:

[0014]

[0015] The molar ratio of 4-(hydroxymethyl)phenylboronic acid pinacol ester to N,N'-carbonyldiimidazole is preferably 1:(1.5-2.5), more preferably 1:2.

[0016] The solvent for the reaction is preferably dichloromethane.

[0017] The reaction temperature is preferably 25-35°C, more preferably 30°C.

[0018] The reaction time is preferably 10 to 15 hours, more preferably 12 hours.

[0019] The molar ratio of paclitaxel to the N,N'-carbonyldiimidazole-modified 4-(hydroxymethyl)phenylboronic acid pinacol ester shown in the structure of formula III is preferably 1:(1.5-2.5), and more preferably 1:2.

[0020] Preferably, the reaction of paclitaxel with N,N'-carbonyldiimidazole-modified 4-(hydroxymethyl)phenylboronic acid pinacol ester shown in the structure of formula III is carried out under the action of a catalyst.

[0021] The catalyst is preferably 4-dimethylaminopyridine (DMAP).

[0022] The amount of the catalyst added is preferably 1 to 3 times the molar amount of paclitaxel, more preferably 2 times.

[0023] The reaction temperature is preferably 25-35°C, more preferably 30°C.

[0024] The reaction time is preferably 10 to 15 hours, more preferably 12 hours.

[0025] Preferably, after the reactive oxygen-responsive paclitaxel prodrug shown in Formula I is obtained by reaction, it is purified by column chromatography. The elution solvent of the column chromatography is preferably a mixed solvent of dichloromethane and ethyl acetate. The volume ratio of dichloromethane to ethyl acetate is preferably 6:1.

[0026] The present invention also provides a method for preparing an active oxygen-responsive paclitaxel prodrug shown in formula II, comprising the following steps:

[0027] S1) reacting paclitaxel with N,N'-carbonyldiimidazole-modified 4-(hydroxymethyl)phenylboronic acid pinacol ester as shown in formula III to obtain an active oxygen-responsive paclitaxel prodrug as shown in formula I;

[0028] S2) hydrolyzing the phenylboronic acid ester bond in the active oxygen responsive paclitaxel prodrug structure of formula I to obtain the active oxygen responsive paclitaxel prodrug of formula II;

[0029]

[0030] The conditions of step S1) are the same as above and will not be repeated here.

[0031] Preferably, in the step S2), the active oxygen-responsive paclitaxel prodrug shown in formula I is reacted under the action of sodium periodate and ammonium acetate (or other reagents that can promote the hydrolysis of phenylboronic acid esters such as citric acid solution) to obtain the active oxygen-responsive paclitaxel prodrug shown in formula II.

[0032] The molar ratio of the active oxygen-responsive paclitaxel prodrug represented by formula I to sodium periodate and ammonium acetate is preferably 1:(1-1.5):(1-1.5), and more preferably 1:1.2:1.2.

[0033] The solvent for the reaction is preferably acetone.

[0034] The reaction temperature is preferably 25-35°C, more preferably 30°C.

[0035] The reaction time is preferably 10 to 15 hours, more preferably 12 hours.

[0036] Preferably, after the reactive oxygen-responsive paclitaxel prodrug shown in Formula II is obtained by reaction, it is purified by column chromatography. The elution solvent of the column chromatography is preferably a mixed solvent of dichloromethane and ethyl acetate. The volume ratio of dichloromethane to ethyl acetate is preferably 3:1.

[0037] The structure of the prepared paclitaxel prodrug is verified by nuclear magnetic resonance.

[0038] The present invention also provides an active oxygen-responsive paclitaxel prodrug nanoparticle, comprising the active oxygen-responsive paclitaxel prodrug.

[0039] Preferably, the nanoparticles also include a polymer carrier.

[0040] The addition of polymer carriers can help further increase the drug loading to about 83%, and can improve the stability of nanoparticles, making them more suitable for in vivo applications.

[0041] The present invention has no particular limitation on the polymer carrier, and any generally applicable carrier known to those skilled in the art can be used.

[0042] Preferably, the polymer carrier includes one or more of polyethylene glycol-poly (lactide-co-glycolide) (PEG-PLGA), polyethylene glycol-phosphatidylethanolamine (PEG-DSPE), polyethylene glycol-polylactic acid copolymer (PEG-PDLLA), polyethylene glycol-polycaprolactone (PEG-PCL), methoxy polyethylene glycol-poly DL-lactide (mPEG-PDLLA), etc.

[0043] The ratio of the paclitaxel prodrug to the polymer carrier can be adjusted as needed.

[0044] Preferably, the molar ratio of the paclitaxel prodrug to the polymer carrier is 9:1 to 1:9, more preferably 1:1 to 1:5, and most preferably 1:5. In some specific embodiments of the present invention, the molar ratio of the paclitaxel prodrug to the polymer carrier is 9:1, 5:1, 1:1, 1:5, 1:9, or a range value with the above ratios as the upper or lower limit.

[0045] Within the above ratio range, the active oxygen-responsive paclitaxel prodrug nanoparticles can obtain an optimal drug loading amount and drug loading efficiency.

[0046] The present invention provides a method for preparing the above-mentioned active oxygen-responsive paclitaxel prodrug nanoparticles, comprising the following steps:

[0047] The active oxygen-responsive paclitaxel prodrug is dissolved in a solvent and then dropped into water to obtain active oxygen-responsive paclitaxel prodrug nanoparticles.

[0048] The solvent is preferably an organic solvent such as N,N-dimethylformamide, dimethyl sulfoxide or methanol.

[0049] The concentration of the paclitaxel prodrug in the solvent is preferably 3 to 8 mg / 100 μL.

[0050] Preferably, the solution of the above active oxygen-responsive paclitaxel prodrug is dropped into vigorously stirred water to obtain active oxygen-responsive paclitaxel prodrug nanoparticles.

[0051] During the dropwise addition process, keeping the aqueous phase in a vigorous stirring state is more conducive to the generation of nanoparticles.

[0052] The volume of the water is preferably 12-18 times, more preferably 10 times, the volume of the paclitaxel prodrug solution.

[0053] When the nanoparticles include a polymer carrier, preferably, the solution of the active oxygen responsive paclitaxel prodrug also includes a polymer carrier.

[0054] The present invention provides the use of the above active oxygen-responsive paclitaxel prodrug or the above active oxygen-responsive paclitaxel prodrug nanoparticles in the preparation of anti-tumor drugs.

[0055] In some specific embodiments of the present invention, the tumor is one or more of melanoma, breast cancer, colon cancer, ovarian cancer, head and neck cancer and lung cancer.

[0056] Compared with the prior art, the present invention provides an active oxygen-responsive paclitaxel prodrug having a structure shown in Formula I or Formula II.

[0057] The invention aims at the problem of hydrophobic aggregation of paclitaxel, and modifies the paclitaxel drug. The water solubility of the modified paclitaxel drug is greatly enhanced, and the self-assembly performance is obtained. The nanoparticles can be self-assembled in an aqueous solution to obtain nanoparticles. The nanoparticles have ultra-high drug loading and drug loading efficiency, and can specifically respond to high concentrations of active oxygen in the tumor microenvironment, which can not only improve the insufficient drug loading of traditional paclitaxel nanoparticles, but also respond to and release paclitaxel at the tumor position, which is conducive to reducing the dosage of the drug, reducing toxic and side effects, and has important significance for improving the prognosis of patients and prolonging the survival period of patients. In addition, the preparation method of the nanoparticle is simple and easy, and a simple nanoprecipitation method can be used to achieve a significant improvement in the drug loading and loading efficiency of paclitaxel. Even when the drug loading exceeds 83%, drug precipitation will not occur. The obtained nanoparticles have good stability and uniform particle size distribution, low cost, and are suitable for batch production. BRIEF DESCRIPTION OF THE DRAWINGS

[0058] Figure 1 For PBE-CI 1 H NMR spectrum;

[0059] Figure 2 For paclitaxel-PBE 1 H NMR spectrum;

[0060] Figure 3 For paclitaxel-PBA 1 H NMR spectrum;

[0061] Figure 4 is the particle size distribution diagram of paclitaxel-PBE nanoparticles in water;

[0062] Figure 5 is the particle size distribution diagram of paclitaxel-PBA nanoparticles in water;

[0063] Figure 6 is the reactive oxygen species response release diagram of paclitaxel-PBE;

[0064] Figure 7 is the cytotoxicity graph of paclitaxel-PBE;

[0065] Figure 8 is the cytotoxicity graph of paclitaxel-PBA;

[0066] Fig. 9 The particle size distribution diagram of paclitaxel-PBE and its polymer nanoformulation in water in Examples 6 and 8. DETAILED DESCRIPTION

[0068] To further illustrate the present invention, the active oxygen-responsive paclitaxel prodrug and its high-drug loading nanoformulation and preparation method provided by the present invention are described in detail below in conjunction with the examples. However, it should be understood that these descriptions are only for further illustrating the features and advantages of the present invention, and are not intended to limit the claims of the invention.

[0069] All raw materials of the present invention have no particular limitation on their sources, and can be purchased from the market or prepared according to conventional methods known to those skilled in the art.

[0070] Example 1

[0071] 4-(Hydroxymethyl)phenylboronic acid pinacol ester (PBE-OH) and N,N'-carbonyldiimidazole (CDI) were dissolved in dichloromethane (DCM) at a molar ratio of 1:2 and reacted at 30°C for 12 hours. After the reaction was completed, ethyl acetate was added to the reaction solution for dilution, extracted with ultrapure water for 6 times, and dried to obtain the product PBE-CI with a reaction yield of about 95%. Its structure was verified by nuclear magnetic resonance, and the hydrogen spectrum was as follows Figure 1 shown.

[0072] Paclitaxel and PBE-CI were dissolved in dichloromethane at a molar ratio of 1:2, and 4-dimethylaminopyridine (DMAP) was added as a catalyst at twice the amount of paclitaxel, and the reaction was carried out at 30°C for 12 hours. After the reaction was completed, ethyl acetate was added to the reaction solution for dilution, and the solution was extracted with ultrapure water for 3 times and dried to obtain a crude product, which was then separated by silica gel column chromatography (the eluent was a mixed solvent of dichloromethane and ethyl acetate in a volume ratio of 6:1) to obtain the product paclitaxel-PBE, with a reaction yield of about 70%. Its structure was verified by nuclear magnetic resonance, and the hydrogen spectrum was as shown in FIG. Figure 2 shown.

[0073] Paclitaxel-PBE, sodium periodate (NaIO4) and ammonium acetate (NH4Ac) with a molar ratio of 1:1.2:1.2 were dissolved in acetone and reacted at 30°C for 12 hours. After the reaction was completed, the acetone was drained, extracted with dichloromethane + ultrapure water for 3 times, and drained to obtain a crude product, which was then separated by silica gel column chromatography (the eluent was a mixed solvent of dichloromethane and ethyl acetate in a volume ratio of 3:1) to obtain the product paclitaxel-PBA, with a reaction yield of about 80%. Its structure was verified by nuclear magnetic resonance, and the hydrogen spectrum was as shown in Figure 3 shown.

[0074] The reaction route is as follows:

[0075]

[0076] Example 2

[0077] The paclitaxel-PBE and paclitaxel-PBA nanoparticles were prepared by nanoprecipitation method. The specific method is as follows:

[0078] Dissolve 5 mg of prodrug in 100 μL N,N-dimethylformamide (DMF), add the dissolved drug DMF dropwise into 10 times the volume of ultrapure water with vigorous stirring, and then dialyze with ultrapure water to remove DMF to obtain clear and transparent prodrug nanoparticles. The particle size of the two nanoparticles was tested by Malvern particle size analyzer. The particle size distribution diagram is shown in the figure. Figure 4 and Figure 5 shown.

[0079] Figure 4 and Figure 5 It shows that after the two prodrugs are added to water through the nanoprecipitation method, they can form nanoparticles with uniform particle size and size distribution of about 100 to 200 nm, rather than producing precipitation, aggregation and precipitation.

[0080] Example 3 Paclitaxel-PBE and Paclitaxel-PBA Reactive Oxygen Response Test

[0081] Using high performance liquid chromatography, the response and fragmentation of paclitaxel-PBE under the conditions of simulating the tumor microenvironment (100 μM hydrogen peroxide) are shown in the following figure: Figure 6 After 24 h of treatment, 90% of paclitaxel-PBE was converted to paclitaxel.

[0082] Paclitaxel-PBE and Paclitaxel-PBA release paclitaxel in response to reactive oxygen species in the same manner. The specific mechanism is as follows:

[0083] Reactive oxygen species responsive release mechanism of paclitaxel-PBE:

[0084]

[0085] Mechanism of reactive oxygen species release of paclitaxel-PBA:

[0086]

[0087] Example 4 Cell activity test

[0088] The cytotoxicity of paclitaxel-PBE and paclitaxel-PBA in three tumor cell lines, B16, 4T1 and MC38, was tested. The three cultured cells were planted in 96-well plates at a number of 5000 / well (3 replicates per group). After culturing for 24 hours, paclitaxel-PBE or paclitaxel-PBA solutions with concentrations of 0.0001, 0.001, 0.01, 0.1, 1, 10, 100 μM were added to the wells. After 24 hours of drug action, CCK-8 detection reagent (10%) was added to each well, and the absorbance at 450 nm was detected by an ELISA reader after incubation at 37°C for 0.5 hours.

[0089] The results are as follows Figure 7 , Figure 8 shown.

[0090] Depend on Figure 7 and Figure 8 The results show that the paclitaxel prodrug does not change its ability to kill tumor cells after modification, and the two prodrugs have similar cytotoxicity to paclitaxel.

[0091] Example 5

[0092] By nanoprecipitation, a nanoformulation was prepared with polyethylene glycol-poly(lactide-co-glycolide) (PEG-PLGA) as a carrier, paclitaxel-PBE as a target loaded drug, and a carrier-drug ratio of 9:1. The specific method is as follows:

[0093] Dissolve 5mg of prodrug and different proportions of carrier in 100μL N,N-dimethylformamide (DMF), add the dissolved drug and carrier DMF drop by drop into 10 times the volume of ultrapure water with vigorous stirring, then dialyze to remove DMF with ultrapure water to obtain clear and transparent prodrug nanoparticles. The particle size of the nanoparticles was tested by Malvern particle size analyzer, and the particle size was between 100 and 200nm. The drug loading and drug loading efficiency of different nanoformulations were determined by high performance liquid chromatography, and the drug loading efficiency of paclitaxel-PBE nanoparticles was greater than 99%.

[0094] Example 6

[0095] A nano-preparation was prepared by nanoprecipitation method with PEG-PLGA as carrier, paclitaxel-PBE as target loaded drug, and a carrier to drug ratio of 5:1. The specific method was similar to that of Example 5.

[0096] The particle size of the nanoparticles was tested by Malvern particle size analyzer, and the particle size was between 100 and 200 nm. The drug loading and drug loading efficiency of different nanoformulations were determined by high performance liquid chromatography, and the drug loading efficiency of paclitaxel-PBE nanoparticles was greater than 99%. Fig. 9 shown.

[0097] Example 7

[0098] A nano-preparation was prepared by nanoprecipitation method with PEG-PLGA as carrier, paclitaxel-PBE as target loaded drug, and a carrier-drug ratio of 1:1. The specific method was similar to that of Example 5.

[0099] The particle size of nanoparticles was tested by Malvern particle size analyzer, and the particle size was between 100 and 200 nm. The drug loading and drug loading efficiency of different nanoparticles were determined by high performance liquid chromatography, and the drug loading efficiency of paclitaxel-PBE nanoparticles was greater than 99%.

[0100] Example 8

[0101] A nano-preparation was prepared by nano-precipitation method with PEG-PLGA as carrier, paclitaxel-PBE as target loaded drug, and a carrier to drug ratio of 1:5. The specific method was similar to that of Example 5.

[0102] The particle size of the nanoparticles was tested by Malvern particle size analyzer, and the particle size was between 100 and 200 nm. The drug loading and drug loading efficiency of different nanoformulations were determined by high performance liquid chromatography, and the drug loading efficiency of paclitaxel-PBE nanoparticles was greater than 99%. Fig. 9 shown.

[0103] Example 9

[0104] A nano-preparation was prepared by nano-precipitation method with PEG-PLGA as carrier, paclitaxel-PBE as target loaded drug, and a carrier to drug ratio of 1:9. The specific method was similar to that of Example 5.

[0105] The particle size of nanoparticles was tested by Malvern particle size analyzer, and the particle size was between 100 and 200 nm. The drug loading and drug loading efficiency of different nanoparticles were determined by high performance liquid chromatography, and the drug loading efficiency of paclitaxel-PBE nanoparticles was greater than 99%.

[0106] Example 10

[0107] A nano-preparation was prepared by nanoprecipitation method with PEG-PLGA as carrier, paclitaxel-PBA as target loaded drug, and a carrier-drug ratio of 9:1. The specific method is as follows:

[0108] The particle size of nanoparticles was tested by Malvern particle size analyzer, and the particle size was between 100 and 200 nm. The drug loading and drug loading efficiency of different nanoparticles were determined by high performance liquid chromatography, and the drug loading efficiency of paclitaxel-PBE nanoparticles was greater than 99%.

[0109] Embodiment 11

[0110] A nano-preparation was prepared by nanoprecipitation method with PEG-PLGA as carrier, paclitaxel-PBA as target loaded drug, and a carrier to drug ratio of 5:1. The specific method was similar to that of Example 5.

[0111] The particle size of nanoparticles was tested by Malvern particle size analyzer, and the particle size was between 100 and 200 nm. The drug loading and drug loading efficiency of different nanoparticles were determined by high performance liquid chromatography, and the drug loading efficiency of paclitaxel-PBE nanoparticles was greater than 99%.

[0112] Example 12

[0113] A nano-preparation was prepared by nano-precipitation method with PEG-PLGA as carrier, paclitaxel-PBA as target loaded drug, and a carrier-drug ratio of 1:1. The specific method was similar to that of Example 5.

[0114] The particle size of nanoparticles was tested by Malvern particle size analyzer, and the particle size was between 100 and 200 nm. The drug loading and drug loading efficiency of different nanoparticles were determined by high performance liquid chromatography, and the drug loading efficiency of paclitaxel-PBE nanoparticles was greater than 99%.

[0115] Embodiment 13

[0116] A nano-preparation was prepared by nano-precipitation method with PEG-PLGA as carrier, paclitaxel-PBA as target loaded drug, and a carrier to drug ratio of 1:5. The specific method was similar to that of Example 5.

[0117] The particle size of nanoparticles was tested by Malvern particle size analyzer, and the particle size was between 100 and 200 nm. The drug loading and drug loading efficiency of different nanoparticles were determined by high performance liquid chromatography, and the drug loading efficiency of paclitaxel-PBE nanoparticles was greater than 99%.

[0118] Embodiment 14

[0119] A nano-preparation was prepared by nanoprecipitation method with PEG-PLGA as carrier, paclitaxel-PBA as target loaded drug, and a carrier to drug ratio of 1:9. The specific method was similar to that of Example 5.

[0120] The particle size of nanoparticles was tested by Malvern particle size analyzer, and the particle size was between 100 and 200 nm. The drug loading and drug loading efficiency of different nanoparticles were determined by high performance liquid chromatography, and the drug loading efficiency of paclitaxel-PBE nanoparticles was greater than 99%.

[0121] Embodiment 15

[0122] By nanoprecipitation method, a nanoformulation was prepared with polyethylene glycol-phosphatidylethanolamine (PEG-DSPE) as carrier, paclitaxel-PBE as target loaded drug, and a carrier-drug ratio of 9:1. The specific method is as follows:

[0123] Dissolve 5mg of prodrug and different proportions of carrier in 100μL N,N-dimethylformamide (DMF), add the dissolved drug and carrier DMF drop by drop into 10 times the volume of ultrapure water with vigorous stirring, then dialyze to remove DMF with ultrapure water to obtain clear and transparent prodrug nanoparticles. The particle size of the nanoparticles was tested by Malvern particle size analyzer, and the particle size was between 100 and 200nm. The drug loading and drug loading efficiency of different nanoformulations were determined by high performance liquid chromatography, and the drug loading efficiency of paclitaxel-PBE nanoparticles was greater than 99%.

[0124] Example 16

[0125] A nano-preparation was prepared by nano-precipitation method with PEG-DSPE as carrier, paclitaxel-PBE as target loaded drug, and a carrier to drug ratio of 5:1. The specific method was similar to that of Example 5.

[0126] The particle size of nanoparticles was tested by Malvern particle size analyzer, and the particle size was between 100 and 200 nm. The drug loading and drug loading efficiency of different nanoparticles were determined by high performance liquid chromatography, and the drug loading efficiency of paclitaxel-PBE nanoparticles was greater than 99%.

[0127] Embodiment 17

[0128] A nano-preparation was prepared by nano-precipitation method with PEG-DSPE as carrier, paclitaxel-PBE as target loaded drug, and a carrier-drug ratio of 1:1. The specific method was similar to that of Example 5.

[0129] The particle size of nanoparticles was tested by Malvern particle size analyzer, and the particle size was between 100 and 200 nm. The drug loading and drug loading efficiency of different nanoparticles were determined by high performance liquid chromatography, and the drug loading efficiency of paclitaxel-PBE nanoparticles was greater than 99%.

[0130] Embodiment 18

[0131] A nano-preparation was prepared by nano-precipitation method with PEG-DSPE as carrier, paclitaxel-PBE as target loaded drug, and a carrier to drug ratio of 1:5. The specific method was similar to that of Example 5.

[0132] The particle size of nanoparticles was tested by Malvern particle size analyzer, and the particle size was between 100 and 200 nm. The drug loading and drug loading efficiency of different nanoparticles were determined by high performance liquid chromatography, and the drug loading efficiency of paclitaxel-PBE nanoparticles was greater than 99%.

[0133] Embodiment 19

[0134] A nano-preparation was prepared by nano-precipitation method with PEG-DSPE as carrier, paclitaxel-PBE as target loaded drug, and a carrier to drug ratio of 1:9. The specific method was similar to that of Example 5.

[0135] The particle size of nanoparticles was tested by Malvern particle size analyzer, and the particle size was between 100 and 200 nm. The drug loading and drug loading efficiency of different nanoparticles were determined by high performance liquid chromatography, and the drug loading efficiency of paclitaxel-PBE nanoparticles was greater than 99%.

[0136] Embodiment 20

[0137] A nano-preparation was prepared by nanoprecipitation method with PEG-DSPE as carrier and paclitaxel-PBA as target loaded drug, with the carrier and drug ratio of 9:1. The specific method is as follows:

[0138] The particle size of nanoparticles was tested by Malvern particle size analyzer, and the particle size was between 100 and 200 nm. The drug loading and drug loading efficiency of different nanoparticles were determined by high performance liquid chromatography, and the drug loading efficiency of paclitaxel-PBE nanoparticles was greater than 99%.

[0139] Embodiment 21

[0140] A nano-preparation was prepared by nano-precipitation method with PEG-DSPE as carrier, paclitaxel-PBA as target loaded drug, and a carrier to drug ratio of 5:1. The specific method was similar to that of Example 5.

[0141] The particle size of nanoparticles was tested by Malvern particle size analyzer, and the particle size was between 100 and 200 nm. The drug loading and drug loading efficiency of different nanoparticles were determined by high performance liquid chromatography, and the drug loading efficiency of paclitaxel-PBE nanoparticles was greater than 99%.

[0142] Embodiment 22

[0143] A nano-preparation was prepared by nano-precipitation method with PEG-DSPE as carrier, paclitaxel-PBA as target loaded drug, and a carrier-drug ratio of 1:1. The specific method was similar to that of Example 5.

[0144] The particle size of nanoparticles was tested by Malvern particle size analyzer, and the particle size was between 100 and 200 nm. The drug loading and drug loading efficiency of different nanoparticles were determined by high performance liquid chromatography, and the drug loading efficiency of paclitaxel-PBE nanoparticles was greater than 99%.

[0145] Embodiment 23

[0146] A nano-preparation was prepared by nano-precipitation method with PEG-DSPE as carrier, paclitaxel-PBA as target loaded drug, and a carrier to drug ratio of 1:5. The specific method was similar to that of Example 5.

[0147] The particle size of nanoparticles was tested by Malvern particle size analyzer, and the particle size was between 100 and 200 nm. The drug loading and drug loading efficiency of different nanoparticles were determined by high performance liquid chromatography, and the drug loading efficiency of paclitaxel-PBE nanoparticles was greater than 99%.

[0148] Embodiment 24

[0149] A nano-preparation was prepared by nano-precipitation method with PEG-DSPE as carrier, paclitaxel-PBA as target loaded drug, and a carrier to drug ratio of 1:9. The specific method was similar to that of Example 5.

[0150] The particle size of nanoparticles was tested by Malvern particle size analyzer, and the particle size was between 100 and 200 nm. The drug loading and drug loading efficiency of different nanoparticles were determined by high performance liquid chromatography, and the drug loading efficiency of paclitaxel-PBE nanoparticles was greater than 99%.

[0151] Embodiment 25

[0152] A nanoformulation was prepared by nanoprecipitation method with polyethylene glycol-polycaprolactone (PEG-PCL) as carrier, paclitaxel-PBE as target loaded drug, and a carrier-drug ratio of 9:1. The specific method is as follows:

[0153] Dissolve 5mg of prodrug and different proportions of carrier in 100μL N,N-dimethylformamide (DMF), add the dissolved drug and carrier DMF drop by drop into 10 times the volume of ultrapure water with vigorous stirring, then dialyze to remove DMF with ultrapure water to obtain clear and transparent prodrug nanoparticles. The particle size of the nanoparticles was tested by Malvern particle size analyzer, and the particle size was between 100 and 200nm. The drug loading and drug loading efficiency of different nanoformulations were determined by high performance liquid chromatography, and the drug loading efficiency of paclitaxel-PBE nanoparticles was greater than 99%.

[0154] Embodiment 26

[0155] A nano-preparation was prepared by nanoprecipitation method with PEG-PCL as carrier, paclitaxel-PBE as target loaded drug, and a carrier to drug ratio of 5:1. The specific method was similar to that of Example 5.

[0156] The particle size of nanoparticles was tested by Malvern particle size analyzer, and the particle size was between 100 and 200 nm. The drug loading and drug loading efficiency of different nanoparticles were determined by high performance liquid chromatography, and the drug loading efficiency of paclitaxel-PBE nanoparticles was greater than 99%.

[0157] Embodiment 27

[0158] A nano-preparation was prepared by nanoprecipitation method with PEG-PCL as carrier, paclitaxel-PBE as target loaded drug, and a carrier-drug ratio of 1:1. The specific method was similar to that of Example 5.

[0159] The particle size of nanoparticles was tested by Malvern particle size analyzer, and the particle size was between 100 and 200 nm. The drug loading and drug loading efficiency of different nanoparticles were determined by high performance liquid chromatography, and the drug loading efficiency of paclitaxel-PBE nanoparticles was greater than 99%.

[0160] Embodiment 28

[0161] A nano-preparation was prepared by nano-precipitation method with PEG-PCL as carrier, paclitaxel-PBE as target loaded drug, and a carrier to drug ratio of 1:5. The specific method was similar to that of Example 5.

[0162] The particle size of nanoparticles was tested by Malvern particle size analyzer, and the particle size was between 100 and 200 nm. The drug loading and drug loading efficiency of different nanoparticles were determined by high performance liquid chromatography, and the drug loading efficiency of paclitaxel-PBE nanoparticles was greater than 99%.

[0163] Embodiment 29

[0164] A nano-preparation was prepared by nanoprecipitation method with PEG-PCL as carrier, paclitaxel-PBE as target loaded drug, and a carrier to drug ratio of 1:9. The specific method was similar to that of Example 5.

[0165] The particle size of nanoparticles was tested by Malvern particle size analyzer, and the particle size was between 100 and 200 nm. The drug loading and drug loading efficiency of different nanoparticles were determined by high performance liquid chromatography, and the drug loading efficiency of paclitaxel-PBE nanoparticles was greater than 99%.

[0166] Embodiment 30

[0167] A nano-preparation was prepared by nanoprecipitation method with PEG-PCL as carrier and paclitaxel-PBA as target loaded drug, with the carrier and drug ratio of 9:1. The specific method is as follows:

[0168] The particle size of nanoparticles was tested by Malvern particle size analyzer, and the particle size was between 100 and 200 nm. The drug loading and drug loading efficiency of different nanoparticles were determined by high performance liquid chromatography, and the drug loading efficiency of paclitaxel-PBE nanoparticles was greater than 99%.

[0169] Embodiment 31

[0170] A nano-preparation was prepared by nano-precipitation method with PEG-PCL as carrier, paclitaxel-PBA as target loaded drug, and a carrier to drug ratio of 5:1. The specific method was similar to that of Example 5.

[0171] The particle size of nanoparticles was tested by Malvern particle size analyzer, and the particle size was between 100 and 200 nm. The drug loading and drug loading efficiency of different nanoparticles were determined by high performance liquid chromatography, and the drug loading efficiency of paclitaxel-PBE nanoparticles was greater than 99%.

[0172] Embodiment 32

[0173] A nano-preparation was prepared by nano-precipitation method with PEG-PCL as carrier, paclitaxel-PBA as target loaded drug, and a carrier-drug ratio of 1:1. The specific method was similar to that of Example 5.

[0174] The particle size of nanoparticles was tested by Malvern particle size analyzer, and the particle size was between 100 and 200 nm. The drug loading and drug loading efficiency of different nanoparticles were determined by high performance liquid chromatography, and the drug loading efficiency of paclitaxel-PBE nanoparticles was greater than 99%.

[0175] Embodiment 33

[0176] A nano-preparation was prepared by nano-precipitation method with PEG-PCL as carrier, paclitaxel-PBA as target loaded drug, and a carrier to drug ratio of 1:5. The specific method was similar to that of Example 5.

[0177] The particle size of nanoparticles was tested by Malvern particle size analyzer, and the particle size was between 100 and 200 nm. The drug loading and drug loading efficiency of different nanoparticles were determined by high performance liquid chromatography, and the drug loading efficiency of paclitaxel-PBE nanoparticles was greater than 99%.

[0178] Embodiment 34

[0179] A nano-preparation was prepared by nano-precipitation method with PEG-PCL as carrier, paclitaxel-PBA as target loaded drug, and a carrier to drug ratio of 1:9. The specific method was similar to that of Example 5.

[0180] The particle size of nanoparticles was tested by Malvern particle size analyzer, and the particle size was between 100 and 200 nm. The drug loading and drug loading efficiency of different nanoparticles were determined by high performance liquid chromatography, and the drug loading efficiency of paclitaxel-PBE nanoparticles was greater than 99%.

[0181] Embodiment 35

[0182] Nanoformulations were prepared by nanoprecipitation method with methoxypolyethylene glycol-poly DL-lactide (mPEG-PDLLA) as carrier, paclitaxel-PBE as target loaded drug, and a carrier-drug ratio of 9:1.

[0183] The specific method is as follows:

[0184] Dissolve 5mg of prodrug and different proportions of carrier in 100μL N,N-dimethylformamide (DMF), add the dissolved drug and carrier DMF drop by drop into 10 times the volume of ultrapure water with vigorous stirring, then dialyze to remove DMF with ultrapure water to obtain clear and transparent prodrug nanoparticles. The particle size of the nanoparticles was tested by Malvern particle size analyzer, and the particle size was between 100 and 200nm. The drug loading and drug loading efficiency of different nanoformulations were determined by high performance liquid chromatography, and the drug loading efficiency of paclitaxel-PBE nanoparticles was greater than 99%.

[0185] Embodiment 36

[0186] A nano-preparation was prepared by nanoprecipitation method with mPEG-PDLLA as carrier, paclitaxel-PBE as target loaded drug, and a carrier to drug ratio of 5:1. The specific method was similar to that of Example 5.

[0187] The particle size of nanoparticles was tested by Malvern particle size analyzer, and the particle size was between 100 and 200 nm. The drug loading and drug loading efficiency of different nanoparticles were determined by high performance liquid chromatography, and the drug loading efficiency of paclitaxel-PBE nanoparticles was greater than 99%.

[0188] Embodiment 37

[0189] A nano-preparation was prepared by nanoprecipitation method with mPEG-PDLLA as carrier, paclitaxel-PBE as target loaded drug, and a carrier-drug ratio of 1:1. The specific method was similar to that of Example 5.

[0190] The particle size of nanoparticles was tested by Malvern particle size analyzer, and the particle size was between 100 and 200 nm. The drug loading and drug loading efficiency of different nanoparticles were determined by high performance liquid chromatography, and the drug loading efficiency of paclitaxel-PBE nanoparticles was greater than 99%.

[0191] Embodiment 38

[0192] The nano-preparation was prepared by nanoprecipitation method with mPEG-PDLLA as carrier, paclitaxel-PBE as target loaded drug, and the ratio of carrier to drug being 1:5. The specific method was similar to that of Example 5.

[0193] The particle size of nanoparticles was tested by Malvern particle size analyzer, and the particle size was between 100 and 200 nm. The drug loading and drug loading efficiency of different nanoparticles were determined by high performance liquid chromatography, and the drug loading efficiency of paclitaxel-PBE nanoparticles was greater than 99%.

[0194] Embodiment 39

[0195] By nanoprecipitation method, a nanoformulation was prepared with mPEG-PDLLA as carrier, paclitaxel-PBE as target loaded drug, and a carrier to drug ratio of 1:9. The specific method was similar to that of Example 5.

[0196] The particle size of nanoparticles was tested by Malvern particle size analyzer, and the particle size was between 100 and 200 nm. The drug loading and drug loading efficiency of different nanoparticles were determined by high performance liquid chromatography, and the drug loading efficiency of paclitaxel-PBE nanoparticles was greater than 99%.

[0197] Embodiment 40

[0198] By nanoprecipitation method, a nanoformulation was prepared with mPEG-PDLLA as carrier, paclitaxel-PBA as target loaded drug, and a carrier-drug ratio of 9:1. The specific method is as follows:

[0199] The particle size of nanoparticles was tested by Malvern particle size analyzer, and the particle size was between 100 and 200 nm. The drug loading and drug loading efficiency of different nanoparticles were determined by high performance liquid chromatography, and the drug loading efficiency of paclitaxel-PBE nanoparticles was greater than 99%.

[0200] Embodiment 41

[0201] By nanoprecipitation method, a nanoformulation was prepared with mPEG-PDLLA as carrier, paclitaxel-PBA as target loaded drug, and a carrier to drug ratio of 5:1. The specific method was similar to that of Example 5.

[0202] The particle size of nanoparticles was tested by Malvern particle size analyzer, and the particle size was between 100 and 200 nm. The drug loading and drug loading efficiency of different nanoparticles were determined by high performance liquid chromatography, and the drug loading efficiency of paclitaxel-PBE nanoparticles was greater than 99%.

[0203] Embodiment 42

[0204] The nano-preparation was prepared by nanoprecipitation method with mPEG-PDLLA as carrier, paclitaxel-PBA as target loaded drug, and the ratio of carrier to drug being 1:1. The specific method was similar to that of Example 5.

[0205] The particle size of nanoparticles was tested by Malvern particle size analyzer, and the particle size was between 100 and 200 nm. The drug loading and drug loading efficiency of different nanoparticles were determined by high performance liquid chromatography, and the drug loading efficiency of paclitaxel-PBE nanoparticles was greater than 99%.

[0206] Embodiment 43

[0207] The nano-preparation was prepared by nano-precipitation method with mPEG-PDLLA as carrier, paclitaxel-PBA as target loaded drug, and the ratio of carrier to drug being 1:5. The specific method was similar to that of Example 5.

[0208] The particle size of nanoparticles was tested by Malvern particle size analyzer, and the particle size was between 100 and 200 nm. The drug loading and drug loading efficiency of different nanoparticles were determined by high performance liquid chromatography, and the drug loading efficiency of paclitaxel-PBE nanoparticles was greater than 99%.

[0209] Embodiment 44

[0210] A nano-preparation was prepared by nanoprecipitation method with mPEG-PDLLA as carrier, paclitaxel-PBA as target loaded drug, and a carrier to drug ratio of 1:9. The specific method was similar to that of Example 5.

[0211] The particle size of nanoparticles was tested by Malvern particle size analyzer, and the particle size was between 100 and 200 nm. The drug loading and drug loading efficiency of different nanoparticles were determined by high performance liquid chromatography, and the drug loading efficiency of paclitaxel-PBE nanoparticles was greater than 99%.

[0212] The above embodiments are only used to help understand the method and core idea of ​​the present invention. It should be noted that, for those skilled in the art, several improvements and modifications can be made to the present invention without departing from the principles of the present invention, and these improvements and modifications also fall within the scope of protection of the claims of the present invention.

Claims

1. An active oxygen-responsive paclitaxel prodrug having a structure shown in Formula I or Formula II:

2. A method for preparing an active oxygen-responsive paclitaxel prodrug as shown in formula I, comprising the following steps: The paclitaxel is reacted with the N,N'-carbonyldiimidazole-modified 4-(hydroxymethyl)phenylboronic acid pinacol ester shown in the structure of formula III to obtain the active oxygen-responsive paclitaxel prodrug shown in formula I; 3. A method for preparing an active oxygen-responsive paclitaxel prodrug as shown in formula II, comprising the following steps: S1) reacting paclitaxel with N,N'-carbonyldiimidazole-modified 4-(hydroxymethyl)phenylboronic acid pinacol ester as shown in formula III to obtain an active oxygen-responsive paclitaxel prodrug as shown in formula I; S2) hydrolyzing the phenylboronic acid ester bond in the active oxygen responsive paclitaxel prodrug structure of formula I to obtain the active oxygen responsive paclitaxel prodrug of formula II; 4. The preparation method according to claim 2 or 3, characterized in that: The N,N'-carbonyldiimidazole-modified 4-(hydroxymethyl)phenylboronic acid pinacol ester shown in the structure of formula III is obtained by reacting 4-(hydroxymethyl)phenylboronic acid pinacol ester and N,N'-carbonyldiimidazole. 5 . An active oxygen-responsive paclitaxel prodrug nanoparticle, comprising the active oxygen-responsive paclitaxel prodrug according to claim 1 .

6. The active oxygen-responsive paclitaxel prodrug nanoparticles according to claim 5, characterized in that: Also included are polymer carriers.

7. A method for preparing active oxygen-responsive paclitaxel prodrug nanoparticles, comprising the following steps: The active oxygen-responsive paclitaxel prodrug of claim 1 is dissolved in a solvent and then dripped into water to obtain active oxygen-responsive paclitaxel prodrug nanoparticles.

8. The preparation method according to claim 7, characterized in that: The N,N-dimethylformamide solution of the active oxygen-responsive paclitaxel prodrug according to claim 1 also includes a polymer carrier.

9. Use of the active oxygen-responsive paclitaxel prodrug according to claim 1 or the active oxygen-responsive paclitaxel prodrug nanoparticles according to any one of claims 5 to 6 in the preparation of anti-tumor drugs.

10. The use according to claim 9, characterized in that: The tumor is one or more of melanoma, breast cancer, colon cancer, ovarian cancer, head and neck cancer, and lung cancer.

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