A cabazitaxel prodrug anti-tumor preparation

Abstract

The present invention discloses a cabazitaxel prodrug anti-tumor preparation, which belongs to the field of new excipients and dosage forms for pharmaceutical preparations. The present invention designs and synthesizes cabazitaxel-fatty alcohol small molecule prodrugs containing different fatty alcohol side chains and different connecting chains, according to the general formula (I), (II), and (III), and prepares self-assembled nanoparticles. Results demonstrate that the cabazitaxel-fatty alcohol small molecule prodrug self-assembled nanoparticles can effectively improve the efficacy of cabazitaxel and reduce its toxic side effects. The length of the branched fatty alcohol side chain, the structure of the fatty alcohol side chain, the elemental composition of the connecting chain, and the length of the connecting chain significantly influence the pharmaceutical properties, in vivo fate, and anti-tumor activity of the cabazitaxel prodrug self-assembled nanoparticles. The prodrug self-assembled nanoparticles exhibit higher anti-tumor activity and lower toxicity than cabazitaxel-linear fatty alcohol small molecule prodrug self-assembled nanoparticles. #imgabs0#

Description

Technical field:

[0001] The present invention belongs to the field of new excipients and new dosage forms for pharmaceutical preparations, and relates to a cabazitaxel prodrug anti-tumor preparation, and specifically to the construction of a cabazitaxel-branched fatty alcohol prodrug and self-assembled nanoparticles thereof, as well as their application in a drug delivery system. Background technology:

[0002] In recent years, the incidence of malignant tumors has been increasing, posing a serious threat to human health. Chemotherapy is one of the most effective strategies for cancer treatment. Cabazitaxel (CTX) is a taxane anti-tumor drug with strong cytotoxicity and anti-tumor effects. However, cabazitaxel can cause serious adverse reactions, including gastrointestinal dysfunction, allergic reactions, renal failure, and neutropenia. Cabazitaxel has extremely poor solubility in water, and the commercially available cabazitaxel solution is Tween 80 and ethanol must be used as solubilizers. Even with the aid of solubilizers, cabazitaxel solutions have poor stability, prone to precipitation upon dilution, and exhibit poor pharmacokinetic properties. These shortcomings limit the clinical application of cabazitaxel.

[0003] Prodrug strategies are an effective method for improving the delivery efficiency of chemotherapy drugs. Structural modification of cabazitaxel through prodrug strategies can effectively improve cabazitaxel's poor solubility and significant toxic side effects. Nanodrug delivery systems can effectively prolong drug circulation in the body and enhance anti-tumor effects. Therefore, self-assembled nanodrug delivery systems based on prodrug strategies combine the advantages of nanotechnology and prodrug strategies, offering advantages such as high drug loading and the absence of solubilizers, and have been widely studied in recent years.

[0004] Prodrugs typically consist of a parent drug, a linker, and a side chain. The linker connects the parent drug and side chain. To construct prodrugs with self-assembly capabilities, existing cabazitaxel prodrugs mostly use linear fatty acids or fatty alcohols as side chains. Aliphatic side chains can increase the structural flexibility of prodrug molecules, balance intermolecular forces, and promote prodrug self-assembly. We hypothesize that branched fatty alcohols can effectively disrupt the tight packing of prodrug molecules, potentially further enhancing their self-assembly ability. Furthermore, the carbon chain length of the branched fatty alcohol may affect the pharmaceutical properties, in vivo fate, and antitumor efficacy of prodrug self-assembled nanoparticles. Currently, no studies have compared the effects of branched fatty alcohol carbon chain length on prodrug self-assembled nanoparticles, nor have any studies reported the effects of branched and linear fatty alcohols as side chains on prodrug self-assembled nanoparticles.

[0005] The tumor microenvironment differs significantly from that of normal tissue cells. Tumor cells produce large amounts of reactive oxygen species and glutathione, creating a redox-unbalanced tumor microenvironment. Monosulfide, disulfide, and monoselenium bonds all possess redox-sensitive properties, intelligently responding to the high redox state within tumor cells and releasing drugs. Different linkers have varying elemental compositions and redox sensitivities. Therefore, cabazitaxel prodrugs modified with different linkers exhibit distinct pharmaceutical properties, in vivo fate, and antitumor effects. Furthermore, the length of the linker also influences the redox sensitivity of the prodrug self-assembled nanoparticles, which in turn affects their antitumor activity. Summary of the invention:

[0006] The purpose of the present invention is to overcome the deficiencies of the above-mentioned prior art and provide a cabazitaxel prodrug anti-tumor preparation, specifically a cabazitaxel-branched fatty alcohol small molecule prodrug and its nanoparticles, as well as the preparation and use thereof. The nanoparticles are self-assembled nanoparticles and have the advantages of small and uniform particle size, high drug loading, good stability, good anti-tumor effect and good safety.

[0007] The purpose of the present invention is to design and synthesize cabazitaxel prodrugs containing branched fatty alcohol side chains of varying lengths, linear fatty alcohol side chains, and different connecting chains, and to prepare self-assembled nanoparticles thereof. Experimental results show that the length of the branched fatty alcohol side chains, the structure of the fatty alcohol side chains (branched or linear), the elemental composition of the connecting chains, and the length of the connecting chains all affect the antitumor efficacy and safety of the prodrug self-assembled nanoparticles. This invention provides more options for the development of new prodrug self-assembled nanodrug delivery systems, meeting the urgent clinical demand for highly effective and low-toxic chemotherapy preparations.

[0008] To achieve the above object, the present invention adopts the following technical solutions:

[0009] Cabazitaxel-branched fatty alcohol small molecule prodrug or a pharmaceutically acceptable salt thereof, wherein the structure of the cabazitaxel-branched fatty alcohol small molecule prodrug is shown in the following general formulas (I), (II), and (III):

[0010]

[0011]

[0012]

[0013] Wherein, n=1 to 3;

[0014] R is a saturated or unsaturated C3-C 30 Hydrocarbon group, wherein R is a hydrocarbon group containing a branched structure, wherein the branched chain is C1-C 18 Alkyl, C2-C 18Alkenyl or C2-C 18 One or more of alkynyl groups.

[0015] Furthermore, R is a saturated or unsaturated C3-C 24 Hydrocarbon group, wherein R is a hydrocarbon group containing a branched structure, wherein the branched chain is a straight chain C6-C 10 Alkyl, C6-C 10 Alkenyl or C6-C 10 One or more of alkynyl groups.

[0016] Furthermore, R is a saturated or unsaturated C 10 -C 24 Hydrocarbon group, wherein R is a hydrocarbon group containing a branched structure, wherein the branched chain is a straight chain C6-C 10 Alkyl, C6-C 10 Alkenyl or C6-C 10 One or more of alkynyl groups.

[0017] Furthermore, R is C 10 -C 24 Alkyl, wherein R is an alkyl containing a branched structure, wherein the branched chain is a straight chain C6-C 10 alkyl.

[0018] Furthermore, R is a saturated or unsaturated C 16 -C 24 Hydrocarbon group, wherein R is a hydrocarbon group containing a branched structure, wherein the branched chain is a straight chain C6-C 10 alkyl.

[0019] Furthermore, R is C 16 -C 24 Alkyl, wherein R is an alkyl containing a branched structure, wherein the branched chain is a straight chain C6-C 10 alkyl.

[0020] When R is an unsaturated hydrocarbon group, the number of alkenyl groups, alkynyl groups, or the sum of alkenyl groups and alkynyl groups contained in the unsaturated hydrocarbon group is 1-5.

[0021] The branched fatty alcohol is one of 2-hexyl-octanol, 1-heptyl-octanol, 2-hexyl-decanol, 1-butyl-dodecanol, 1-heptyl-nonanol, 1-octyl-nonanol, 2-octyl-decanol, 2-heptyl-undecanol, 1-nonyl-decanol, 2-octyl-dodecanol, 2-decyl-tetradecanol or 2-dodecyl-tetradecanol.

[0022] Preferably, the branched fatty alcohol is 2-hexyl-decanol, 2-heptyl-undecanol, 2-octyl-dodecanol or 2-decyl-tetradecanol.

[0023] Cabazitaxel and the branched fatty alcohol in the cabazitaxel-branched fatty alcohol small molecule prodrug are connected by a dibasic acid as a connecting chain. The dibasic acid is a monothiodibasic acid, a monoselenodibasic acid or a dithiodibasic acid, wherein the monothiodibasic acid is monothiodiacetic acid, monothiodipropionic acid or monothiodibutyric acid; the monoselenodibasic acid is monoselenodiacetic acid, monoselenodipropionic acid or monoselenodibutyric acid; the dithiodibasic acid is 2,2'-dithiodiacetic acid, 3,3'-dithiodipropionic acid or 4,4'-dithiodibutyric acid.

[0024] Specifically, the present invention provides cabazitaxel-2-hexyl-decanol prodrug, cabazitaxel-2-heptyl-undecanol prodrug, cabazitaxel-2-octyl-dodecanol prodrug and cabazitaxel-2-decyl-tetradecanol prodrug, and selects 2,2'-dithiodiacetic acid as the connecting chain. The corresponding prodrugs are named CTX-SS-HD, CTX-SS-HU, CTX-SS-OD and CTX-SS-DT, respectively, and their structural formulas are:

[0025]

[0026]

[0027] The present invention provides a linear fatty alcohol small molecule prodrug - cabazitaxel-arachidyl alcohol prodrug, selecting 2,2'-dithiodiacetic acid as the connecting chain, and the corresponding prodrug is named CTX-SS-AA, and its structural formula is:

[0028]

[0029] Cabazitaxel-arachidyl alcohol prodrug with 2,2'-dithiodiacetic acid as the linker (CTX-SS-AA)

[0030] The present invention also provides a cabazitaxel-2-octyl-dodecanol prodrug with 4,4'-dithiodibutyric acid as a connecting chain. The corresponding prodrug is named γ-CTX-SS-OD, and its structural formula is:

[0031]

[0032] Cabazitaxel-2-octyl-dodecanol prodrug (γ-CTX-SS-OD) with 4,4'-dithiodibutyric acid as the linker

[0033] The present invention also provides cabazitaxel-2-octyl-dodecanol prodrugs using monothiodiacetic acid and monoselenodiacetic acid as connecting chains. The corresponding prodrugs are named CTX-S-OD and CTX-Se-OD, respectively, and their structural formulas are:

[0034]

[0035] The method for synthesizing the cabazitaxel-branched fatty alcohol small molecule prodrug comprises the following steps:

[0036] Step 1: dissolving a dibasic acid into a dibasic acid anhydride, and then subjecting the dibasic acid to an esterification reaction with a branched fatty alcohol to obtain a branched fatty alcohol-dibasic acid monoester intermediate, wherein the molar ratio of the branched fatty alcohol to the dibasic acid anhydride is (1-10):(5-15), and the dibasic acid is a monothiodibasic acid, a monoselenodibasic acid, or a dithiodibasic acid;

[0037] Step 2: A branched fatty alcohol-diacid unilateral ester reacts with cabazitaxel to form an ester to obtain the final product, cabazitaxel-branched fatty alcohol small molecule prodrug, wherein the molar ratio of branched fatty alcohol-diacid unilateral ester: cabazitaxel is 1:(0.5-10), and the reaction equation is as follows:

[0038]

[0039]

[0040] Wherein, n=1 to 3;

[0041] R is a saturated or unsaturated C3-C 30 Hydrocarbon group, wherein R is a hydrocarbon group containing a branched structure, wherein the branched chain is C1-C 18 Alkyl, C2-C 18 Alkenyl or C2-C 18 One or more of alkynyl groups.

[0042] The synthesis method of the above-mentioned cabazitaxel-branched fatty alcohol small molecule prodrug specifically comprises the following steps:

[0043] (1) dissolving the dibasic acid in acetic anhydride and stirring at room temperature for 2-4 hours to convert the dibasic acid into dibasic anhydride. After the reaction is complete, toluene is added and the toluene and acetic anhydride are removed by rotary evaporation under reduced pressure;

[0044] (2) taking a branched fatty alcohol and 4-dimethylaminopyridine (DMAP), dissolving them together with the dibasic acid anhydride obtained in step (1) in dichloromethane, stirring at room temperature for 12-18 hours, and separating by chromatography to obtain an intermediate product: a branched fatty alcohol-dibasic acid monoester;

[0045] (3) 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDCI), 1-hydroxybenzotriazole (HOBt) and 4-dimethylaminopyridine (DMAP) are dissolved together with the intermediate product branched fatty alcohol-diacid monoester in anhydrous dichloromethane, stirred in an ice bath for 2-4 hours, and then cabazitaxel is added. The mixture is stirred at room temperature for 24-48 hours, and then purified by preparative liquid phase separation to obtain the final product: cabazitaxel-branched fatty alcohol small molecule prodrug.

[0046] The synthesis method of the cabazitaxel-branched fatty alcohol small molecule prodrug is carried out under nitrogen protection throughout the reaction process.

[0047] In the step (1), the dibasic acid is monothiodiacetic acid, monothiodipropionic acid, monothiodibutyric acid, monoselenodiacetic acid, monoselenodipropionic acid, monoselenodibutyric acid, 2,2'-dithiodiacetic acid, 3,3'-dithiodipropionic acid or 4,4'-dithiodibutyric acid.

[0048] In the step (1), the ratio of dibasic acid to acetic anhydride is 1:(1-10), preferably 1:(1-2), in units of mmol:ml. In the step (2), the branched fatty alcohol is C3-C 30 Saturated or unsaturated fatty alcohol, the side chain is C1-C 18 Alkyl, C2-C 18 Alkenyl or C2-C 18 One or more of alkynyl groups.

[0049] In the step (2), the molar ratio of DMAP:branched fatty alcohol:dibasic acid anhydride is 1:(1-10):(5-15), preferably 1:(2-5):(10-15).

[0050] In the step (3), the molar ratio of the intermediate product branched fatty alcohol-diacid monoester:HOBt:EDCI:DMAP:cabazitaxel is 1:(1-10):(2-6):(0.2-5):(0.5-10), preferably 1:(1-2):(2-4):(0.5-2):(0.8-2).

[0051] In the step (3), the purity of the prepared cabazitaxel-branched fatty alcohol small molecule prodrug is above 99%.

[0052] The present invention also provides a method for synthesizing monoselenodiacetic acid, comprising the following steps:

[0053] Selenium powder and a small amount of water are added to an eggplant-shaped flask, which is placed in an ice-water bath. An aqueous sodium borohydride solution is slowly added dropwise and stirred until the solution becomes clear. The temperature is raised to 100-110°C and stirred for 30-45 minutes. After cooling to room temperature, an aqueous bromoacetic acid solution is added dropwise. After reacting for 10-12 hours, the reaction solution is filtered and extracted with ethyl acetate until the aqueous layer is colorless. The ethyl acetate layer is dried and then rotary evaporated under reduced pressure to remove the ethyl acetate to obtain the product. The entire reaction is carried out under nitrogen protection.

[0054] The present invention also provides self-assembled nanoparticles of cabazitaxel-fatty alcohol small molecule prodrugs, wherein the prodrug self-assembled nanoparticles are non-PEGylated prodrug self-assembled nanoparticles, PEG-modified / actively targeted modified prodrug self-assembled nanoparticles, or prodrug self-assembled nanoparticles encapsulating hydrophobic fluorescent substances / drugs.

[0055] The method for preparing the cabazitaxel-branched fatty alcohol small molecule prodrug self-assembled nanoparticles comprises the following steps:

[0056] In the case of non-PEGylated cabazitaxel-fatty alcohol small molecule prodrug self-assembled nanoparticles, the preparation method is as follows: a certain amount of prodrug is dissolved in an appropriate amount of organic solvent, the solution is slowly added dropwise to water under stirring, and the prodrug spontaneously forms uniform nanoparticles; the organic solvent in the preparation is removed by reduced pressure rotary evaporation to obtain a nanocolloidal solution free of any organic solvent, namely the non-PEGylated cabazitaxel-fatty alcohol small molecule prodrug.

[0057] The preparation method of PEG-modified / active targeting group-modified cabazitaxel-fatty alcohol small molecule prodrug self-assembled nanoparticles comprises the following steps: dissolving a certain amount of PEG modifier / active targeting modifier and prodrug in an appropriate amount of organic solvent; slowly adding the solution dropwise into water under stirring, so that the prodrug spontaneously forms uniform nanoparticles; and removing the organic solvent in the preparation by rotary evaporation under reduced pressure to obtain a nanocolloidal solution free of any organic solvent, namely, the PEG-modified / active targeting group-modified cabazitaxel-fatty alcohol small molecule prodrug self-assembled nanoparticles. The mass ratio of cabazitaxel-branched fatty alcohol small molecule prodrug to PEG modifier / active targeting modifier is 1:(0.1-1); the PEG modifier is an amphiphilic polymer or targeting group such as DSPE-PEG, TPGS, PLGA-PEG, PE-PEG or DSPE-PEG-FA; and the active targeting modifier is a substance capable of targeting specific tissues, such as an antibody, a sugar residue, a hormone, a receptor or a ligand.

[0058] When the cabazitaxel-fatty alcohol small molecule prodrug self-assembled nanoparticles are loaded with a hydrophobic fluorescent substance / drug, the preparation method is as follows: a certain amount of PEG modifier, hydrophobic fluorescent substance / drug, and prodrug are dissolved in an appropriate amount of organic solvent, the solution is slowly added dropwise to water under stirring, and the prodrug spontaneously forms uniform nanoparticles; the organic solvent in the preparation is removed by reduced pressure rotary evaporation to obtain a nanocolloidal solution free of organic solvent, namely the cabazitaxel-fatty alcohol small molecule prodrug self-assembled nanoparticles loaded with a hydrophobic fluorescent substance / drug, wherein the mass ratio of cabazitaxel-branched fatty alcohol small molecule prodrug to the PEG modifier and the hydrophobic fluorescent substance / drug is 1:(0.1-1):(0.1-1).

[0059] Application of the cabazitaxel-branched fatty alcohol small molecule prodrug or the self-assembled nanoparticles in the preparation of anti-tumor drugs.

[0060] Application of the cabazitaxel-branched fatty alcohol small molecule prodrug or the self-assembled nanoparticles in preparing an injection, oral administration or local administration system.

[0061] Application of the cabazitaxel-branched fatty alcohol small molecule prodrug or the self-assembled nanoparticles in preparing a drug delivery system with improved efficacy and reduced toxicity.

[0062] The lyophilized powder injection of cabazitaxel-branched fatty alcohol small molecule prodrug self-assembled nanoparticles comprises a cabazitaxel-branched fatty alcohol small molecule prodrug self-assembled nanoparticle solution and a lyoprotectant. The concentration of the self-assembled nanoparticle solution is 0.1 mg / mL-20 mg / mL. The lyoprotectant is one or more of monosaccharides such as glucose and galactose, disaccharides such as trehalose and sucrose, polyols such as mannitol, sorbitol, and xylitol, and polymers such as polyethylene glycol, hydroxyethyl starch, or dextran. The amount of the lyoprotectant is 1%-20% (W / V) (i.e., the mass concentration of the lyoprotectant is 10 g / L-200 g / L), preferably 5%-10% (W / V).

[0063] The preparation method of the cabazitaxel-branched fatty alcohol small molecule prodrug self-assembled nanoparticle freeze-dried powder injection comprises the following steps:

[0064] The cabazitaxel-branched fatty alcohol small molecule prodrug self-assembled nanoparticle solution is placed in a vial, and a lyoprotectant is added. After the lyoprotectant is completely dissolved, the solution is pre-frozen at -80°C for 8-12 hours, and then freeze-dried in a freeze dryer for 24-72 hours to obtain a white cake, which is the cabazitaxel-branched fatty alcohol small molecule prodrug self-assembled nanoparticle lyophilized powder injection.

[0065] Beneficial effects of the present invention:

[0066] (1) The present invention designs and synthesizes cabazitaxel-fatty alcohol small molecule prodrugs containing different fatty alcohol side chains and different connecting chains, and the synthesis method is simple and easy; and prepares cabazitaxel-fatty alcohol small molecule prodrug self-assembled nanoparticles with small particle size and uniform particle size distribution, and the preparation method is simple and easy; (2) The effects of four lengths of branched fatty alcohol side chains, the structure of fatty alcohol side chains (branched or linear), and four connecting chains on the pharmaceutical properties, in vivo fate and anti-tumor activity of prodrug self-assembled nanoparticles are investigated. The results showed that cabazitaxel-fatty alcohol small molecule prodrug self-assembled nanoparticles can effectively improve the efficacy of cabazitaxel and reduce its toxic side effects; different side chains and linkers significantly affect the pharmaceutical properties, in vivo fate, and antitumor activity of cabazitaxel prodrug self-assembled nanoparticles; 2-octyl-dodecanol as the side chain has the best safety of prodrug self-assembled nanoparticles; cabazitaxel-branched fatty alcohol small molecule prodrug self-assembled nanoparticles have better antitumor efficacy and safety than cabazitaxel-linear fatty alcohol small molecule prodrug self-assembled nanoparticles; cabazitaxel-fatty alcohol small molecule prodrug self-assembled nanoparticles with disulfide bonds as the linker have a greater advantage in antitumor efficacy; when the linker is 4,4'-dithiodibutyric acid and the side chain is 2-octyl-dodecanol, the compound has the strongest antitumor activity, the best safety, and the highest tolerable dose. This invention provides new strategies and options for the development of highly effective and low-toxic chemotherapy preparations. Description of the drawings:

[0067] Figure 1 This is a diagram showing the changes in tumor volume during the in vivo anti-tumor experiment of the PEG-modified small molecule prodrug self-assembled nanoparticles of Example 13 of the present invention.

[0068] ns: P ≥ 0.05*: P < 0.05**: P < 0.01***: P < 0.001****: P < 0.0001 (all two-sided t-tests)

[0069] Figure 2 This is a graph showing changes in mouse body weight during an in vivo anti-tumor experiment using PEG-modified small molecule prodrug self-assembled nanoparticles according to Example 13 of the present invention.

[0070] ns: P ≥ 0.05*: P < 0.05**: P < 0.01***: P < 0.001****: P < 0.0001 (all two-sided t-tests)

[0071] Figure 3 This is a graph showing routine blood indicators in the in vivo anti-tumor experiment of the PEG-modified small molecule prodrug self-assembled nanoparticles of Example 13 of the present invention.

[0072] Figure 4 This is a diagram showing the changes in tumor volume during the in vivo anti-tumor experiment of the PEG-modified small molecule prodrug self-assembled nanoparticles of Example 14 of the present invention.

[0073] ns: P ≥ 0.05*: P < 0.05**: P < 0.01***: P < 0.001****: P < 0.0001 (all two-sided t-tests)

[0074] Figure 5 This is a graph showing changes in mouse body weight during an in vivo anti-tumor experiment using PEG-modified small molecule prodrug self-assembled nanoparticles according to Example 14 of the present invention.

[0075] ns: P ≥ 0.05*: P < 0.05**: P < 0.01***: P < 0.001****: P < 0.0001 (all two-sided t-tests)

[0076] Figure 6 This is a diagram of tumor burden in the in vivo anti-tumor experiment of the PEG-modified small molecule prodrug self-assembled nanoparticles of Example 14 of the present invention.

[0077] ns: P ≥ 0.05*: P < 0.05**: P < 0.01***: P < 0.001****: P < 0.0001 (all two-sided t-tests)

[0078] Figure 7 This is a graph showing routine blood indicators in the in vivo anti-tumor experiment of the PEG-modified small molecule prodrug self-assembled nanoparticles of Example 14 of the present invention.

[0079] Figure 8 This is a graph showing conventional biochemical indicators in the in vivo anti-tumor experiment of the PEG-modified small molecule prodrug self-assembled nanoparticles of Example 14 of the present invention.

[0080] Figure 9 This is a diagram showing the changes in tumor volume during the in vivo anti-tumor experiment of the PEG-modified small molecule prodrug self-assembled nanoparticles of Example 15 of the present invention.

[0081] ns: P ≥ 0.05*: P < 0.05**: P < 0.01***: P < 0.001****: P < 0.0001 (all two-sided t-tests)

[0082] Figure 10 This is a graph showing changes in mouse body weight during an in vivo anti-tumor experiment using PEG-modified small molecule prodrug self-assembled nanoparticles according to Example 15 of the present invention.

[0083] ns: P ≥ 0.05*: P < 0.05**: P < 0.01***: P < 0.001****: P < 0.0001 (all two-sided t-tests)

[0084] Figure 11This is a diagram of tumor burden in the in vivo anti-tumor experiment of the PEG-modified small molecule prodrug self-assembled nanoparticles of Example 15 of the present invention.

[0085] ns: P ≥ 0.05*: P < 0.05**: P < 0.01***: P < 0.001****: P < 0.0001 (all two-sided t-tests)

[0086] Figure 12 This is a diagram showing the changes in tumor volume during the in vivo anti-tumor experiment of the PEG-modified small molecule prodrug self-assembled nanoparticles of Example 16 of the present invention.

[0087] ns: P ≥ 0.05*: P < 0.05**: P < 0.01***: P < 0.001****: P < 0.0001 (all two-sided t-tests)

[0088] Figure 13 This is a graph showing changes in mouse body weight during an in vivo anti-tumor experiment using PEG-modified small molecule prodrug self-assembled nanoparticles according to Example 16 of the present invention.

[0089] ns: P ≥ 0.05*: P < 0.05**: P < 0.01***: P < 0.001****: P < 0.0001 (all two-sided t-tests)

[0090] Figure 14 This is a diagram of tumor burden in the in vivo anti-tumor experiment of the PEG-modified small molecule prodrug self-assembled nanoparticles of Example 16 of the present invention.

[0091] ns: P ≥ 0.05*: P < 0.05**: P < 0.01***: P < 0.001****: P < 0.0001 (all two-sided t-tests)

[0092] Figure 15 This is a diagram showing the changes in tumor volume during the in vivo anti-tumor experiment of the PEG-modified small molecule prodrug self-assembled nanoparticles of Example 17 of the present invention.

[0093] ns: P ≥ 0.05*: P < 0.05**: P < 0.01***: P < 0.001****: P < 0.0001 (all two-sided t-tests)

[0094] Figure 16 This is a graph showing changes in mouse body weight during an in vivo anti-tumor experiment using PEG-modified small molecule prodrug self-assembled nanoparticles according to Example 17 of the present invention.

[0095] ns: P≥0.05*: P<0.05**: P<0.01***: P<0.001****: P<0.0001 (all two-sided t tests). Specific implementation method:

[0096] The present invention will be further described in detail below with reference to the embodiments.

[0097] Example 1: Synthesis of Cabazitaxel-2-hexyl-decanol Prodrug with 2,2'-dithiodiacetic acid as a Linker

[0098] An appropriate amount of 2,2' dithiodiacetic acid was dissolved in acetic anhydride and placed in a 25 mL eggplant flask. After complete dissolution, the mixture was transferred to a 100 mL eggplant flask under magnetic stirring at 25°C for 2 hours, and three times the amount of toluene was added. The toluene and acetic anhydride were removed by rotary evaporation under reduced pressure. An appropriate amount of dichloromethane was added to dissolve the formed dithiodiacetic anhydride. Then, a dichloromethane solution of 2-hexyl-decanol was added, and a 4-dimethylaminopyridine (DMAP) solution dissolved in dichloromethane was slowly added dropwise. The mixture was magnetically stirred at 25°C for 12 hours. h, obtaining the intermediate product, 2-hexyl-decanol-dithiodiacetic acid monoester, which was separated and purified by column chromatography using a cyclohexane-acetone elution system. The purified product from the previous step was added with a dichloromethane solution of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDCI), 1-hydroxybenzotriazole (HOBt), and 4-dimethylaminopyridine (DMAP). The mixture was activated in an ice bath at 0°C for 2 hours, followed by the addition of a dichloromethane solution of cabazitaxel, and stirred at 25°C for 48 hours. After the reaction, the product was separated using a preparative phase to obtain the cabazitaxel-2-hexyl-decanol prodrug with 2,2'-dithiodiacetic acid as the linker. During the reaction, the ratio of 2,2'-dithiodiacetic acid: acetic anhydride is 1:1, unit: mmol:ml; the molar ratio of DMAP: 2-hexyl-decanol: dithiodiacetic anhydride is 0.4:2:1, and the molar ratio of 2-hexyl-decanol-dithiodiacetic acid monoester: HOBt: EDCI: DMAP: cabazitaxel is 1:1:2:0.4:0.8.

[0099] Using mass spectrometry and 1 H-NMR confirmed the structure of the product. The spectral analysis results are as follows:

[0100] 1H-NMR(600MHz,DMSO-d6)δ7.966(2H,d),7.840(1H,m,),7.650-7.677(2H,m, ),7.348(2H,m),7.174(3H,m),5.812(1H,s,13-H),5.070-5.131(2H,d,2-H,3 '-H),4.954(1H,d,2'-H),4.931(3H,m,3-H,-CH=CH-),4.686(1H,d,4-H),4. 459(1H,m,5-H),4.013(1H,d,20α-H),3.996(4H,m,-OCH2CH2O-),3.857(1H,d ,7-H),3.590(1H,d,8-H),3.291(4H,m,-CH2-SS-CH2-),3.207(4H,m,6α-H,s ,4-COCHCH2),2.650(1H,m,14α-H),2.236-2.495(2H,m,14β-H,15α-H),1.791 (7H,t,6β-H,-CH2CH=CHCH2-),1.504(5H,s,19-H,10-COCH3),1.376(6H,m,-C H2CH2CH2CO-),1.245(31H,t,17-H),0.978(7H,s,16-H),0.852(6H,t,-CH3).

[0101] MS (ESI) m / z for C 65 H 93 NO 17 S2Na[M+Na] + :1246.

[0102] Example 2: Synthesis of Cabazitaxel-2-heptyl-undecanol Prodrug Using 2,2'-dithiodiacetic acid as a Linker

[0103] An appropriate amount of 2,2'-dithiodiacetic acid was dissolved in acetic anhydride and placed in a 25 mL eggplant flask. After complete dissolution, the mixture was transferred to a 100 mL eggplant flask under magnetic stirring at 25°C for 2 hours, and three times the amount of toluene was added. The toluene and acetic anhydride were removed by rotary evaporation under reduced pressure. An appropriate amount of dichloromethane was added to dissolve the formed dithiodiacetic anhydride. Then, a dichloromethane solution of 2-heptyl-undecanol was added, and a 4-dimethylaminopyridine (DMAP) solution dissolved in dichloromethane was slowly added dropwise. The mixture was magnetically stirred at 25°C for 12 hours. h, obtaining the intermediate product, 2-heptyl-undecanol-dithiodiacetic acid monoester, which was separated and purified by column chromatography using a cyclohexane-acetone elution system. The purified product from the previous step was added with a dichloromethane solution of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDCI), 1-hydroxybenzotriazole (HOBt), and 4-dimethylaminopyridine (DMAP). The mixture was activated in an ice bath at 0°C for 2 hours, followed by the addition of a dichloromethane solution of cabazitaxel, and stirred at 25°C for 48 hours. After the reaction, the product was separated using a preparative phase to obtain the cabazitaxel-2-heptyl-undecanol prodrug with 2,2'-dithiodiacetic acid as the linker. During the reaction, the ratio of 2,2'-dithiodiacetic acid: acetic anhydride is 1:1, unit: mmol:ml; the molar ratio of DMAP: 2-heptyl-undecanol: dithiodiacetic anhydride is 0.4:2:1, and the molar ratio of 2-heptyl-undecanol-dithiodiacetic acid monoester: HOBt: EDCI: DMAP: cabazitaxel is 1:1:2:0.4:0.8.

[0104] Using mass spectrometry and 1 H-NMR confirmed the structure of the product. The spectral analysis results are as follows:

[0105] 1H-NMR(600MHz,DMSO-d6)δ7.967(2H,d),7.817(1H,m),7.737(2H,m),7.658 (2H,m),7.175(3H,m),5.814(1H,s,13-H),5.131-5.358(2H,d,2-H,3'-H),5 .071(1H,d,2'-H),4.932(3H,m,3-H,-CH=CH-),4.687(1H,d,4-H),4.460(1 H,m,5-H),4.015(1H,d,20α-H),3.976(4H,m,-OCH2CH2O-),3.857(1H,d,7-H ),3.574(1H,d,8-H),3.293(4H,m,-CH2-SS-CH2-),3.207(4H,m,6α-H,s,4- COCHCH2),2.651(1H,d,18-H),2.237-2.500(2H,m,14β-H,15α-H),1.792(7H ,t,6β-H,-CH2CH=CHCH2-),1.505(5H,s,19-H,10-COCH3),1.377(6H,m,-CH 2CH2CH2CO-),1.243(35H,t,17-H),0.979(7H,s,16-H),0.853(6H,t,-CH3).

[0106] MS (ESI) m / z for C 67 H 97 NO 17 S2Na[M+Na] + :1276.

[0107] Example 3: Synthesis of Cabazitaxel-2-octyl-dodecanol Prodrug Using 2,2'-dithiodiacetic acid as a Linker

[0108] An appropriate amount of 2,2'-dithiodiacetic acid was dissolved in acetic anhydride and placed in a 25 mL eggplant flask. After complete dissolution, the mixture was magnetically stirred at 25°C for 2 hours and then transferred to a 100 mL eggplant flask. Three times the amount of toluene was added. The toluene and acetic anhydride were removed by rotary evaporation under reduced pressure. An appropriate amount of dichloromethane was added to dissolve the formed dithiodiacetic anhydride. Then, a dichloromethane solution of 2-octyl-dodecanol was added, and a dichloromethane solution of 4-dimethylaminopyridine (DMAP) was slowly added dropwise. The mixture was magnetically stirred at 25°C for 12 hours. The intermediate product, 2-octyl-dodecanol-dithiodiacetic acid monoester, was obtained and separated and purified by column chromatography using a cyclohexane-acetone elution system. The purified product from the previous step was added with a dichloromethane solution of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDCI), 1-hydroxybenzotriazole (HOBt), and 4-dimethylaminopyridine (DMAP). The mixture was activated in an ice bath at 0°C for 2 hours, followed by the addition of a dichloromethane solution of cabazitaxel and stirred at 25°C for 48 hours. After the reaction, the product was separated using preparative phase chromatography to obtain the cabazitaxel-2-octyl-dodecanol prodrug with 2,2'-dithiodiacetic acid as the linker. During the reaction, the ratio of 2,2'-dithiodiacetic acid: acetic anhydride is 1:1, unit: mmol:ml; the molar ratio of DMAP: 2-octyl-dodecanol: dithiodiacetic anhydride is 0.4:2:1, and the molar ratio of 2-octyl-dodecanol-dithiodiacetic acid monoester: HOBt: EDCI: DMAP: cabazitaxel is 1:1:2:0.4:0.8.

[0109] Using mass spectrometry and 1 H-NMR confirmed the structure of the product. The spectral analysis results are as follows: 1H-NMR(600MHz,DMSO-d6)δ7.967(2H,t),7.662(3H,d),7.378-7.423(3H,m),7.153-7.189(2H,m),5.807(1H,d,-NH),5.355(1H,d,3'-H),5.124 (1H,d,2-H),5.065(4H,m,2'-H,-CH=CH-,5-H),4.924-4.954(1H,t,7-H ),4.685(1H,d,20α-H),4.477(1H,d,20β-H),3.993-4.012(2H,d,15α-H ,15β-H),3.861(3H,d,3-H),3.736(4H,m,6α-H,4-COCH3),2.647(1H,m,13-H),2.501(3H,t,14α-H,-CH2CO-),1.790(6H,s,-CH2CH=CHCH2-),1. 618(4H,s,18-H),1.503(t,5H,6β-H),1.377(10H,s,16-H,19-H,-CH2CH2CO-),1.240(24H,t,17-H),0.975(3H,t,-CH3),0.850(5H,t,-CH2CH3).

[0110] MS (ESI) m / z for C 69 H 101 NO 17 S2Na[M+Na] + :1303.

[0111] Example 4: Synthesis of Cabazitaxel-2-decyl-tetradecanol Prodrug with 2,2'-dithiodiacetic acid as a Linker

[0112] Dissolve an appropriate amount of 2,2'-dithiodiacetic acid in acetic anhydride and place it in a 25 mL eggplant flask. After complete dissolution, stir magnetically at 25°C for 2 hours and then transfer to a 100 mL eggplant flask. Add three times the amount of toluene and remove the toluene and acetic anhydride by rotary evaporation under reduced pressure. Add an appropriate amount of dichloromethane to dissolve the formed dithiodiacetic anhydride. Then add a dichloromethane solution of 2-decyl-tetradecanol and slowly dropwise add a solution of 4-dimethylaminopyridine (DMAP) dissolved in dichloromethane. Stir magnetically at 25°C for 12 hours. h, obtaining the intermediate product, 2-decyl-tetradecanol-dithiodiacetic acid monoester, which was separated and purified by column chromatography using a cyclohexane-acetone elution system. The purified product from the previous step was added with a dichloromethane solution of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDCI), 1-hydroxybenzotriazole (HOBt), and 4-dimethylaminopyridine (DMAP). The mixture was activated in an ice bath at 0°C for 2 hours, followed by the addition of a dichloromethane solution of cabazitaxel, and stirred at 25°C for 48 hours. After the reaction, the product was separated using a preparative phase to obtain the cabazitaxel-2-decyl-tetradecanol prodrug with 2,2'-dithiodiacetic acid as the linker. During the reaction, the ratio of 2,2'-dithiodiacetic acid: acetic anhydride is 1:1, unit: mmol:ml; the molar ratio of DMAP: 2-decyl-tetradecanol: dithiodiacetic anhydride is 0.4:2:1, and the molar ratio of 2-decyl-tetradecanol-dithiodiacetic acid monoester: HOBt: EDCI: DMAP: cabazitaxel is 1:1:2:0.4:0.8.

[0113] Using mass spectrometry and 1 H-NMR confirmed the structure of the product. The spectral analysis results are as follows:

[0114] 1H-NMR(600MHz,DMSO-d6)δ7.967(2H,d),7.817(1H,m),7.737(2H,m),7.658 (2H,m),7.175(3H,m),5.814(1H,s,13-H),5.131-5.358(2H,d,2-H,3'-H),5 .071(1H,d,2'-H),4.932(3H,m,3-H,-CH=CH-),4.687(1H,d,4-H),4.460(1 H,m,5-H),4.015(1H,d,20α-H),3.976(4H,m,-OCH2CH2O-),3.857(1H,d,7-H ),3.574(1H,d,8-H),3.293(4H,m,-CH2-SS-CH2-),3.207(4H,m,6α-H,s,4- COCHCH2),2.651(1H,d,18-H),2.237-2.500(2H,m,14β-H,15α-H),1.792(7H ,t,6β-H,-CH2CH=CHCH2-),1.505(5H,s,19-H,10-COCH3),1.377(6H,m,-CH 2CH2CH2CO-),1.243(35H,t,17-H),0.979(7H,s,16-H),0.853(6H,t,-CH3).

[0115] MS (ESI) m / z for C 73 H 109 NO 17 S2Na[M+Na] + :1359.

[0116] Example 5: Synthesis of Cabazitaxel-arachidyl alcohol prodrug with 2,2' dithiodiacetic acid as the linker

[0117] Dissolve an appropriate amount of 2,2'-dithiodiacetic acid in acetic anhydride and place in a 25 mL eggplant flask. After complete dissolution, stir magnetically at 25°C for 2 hours and then transfer to a 100 mL eggplant flask. Add three times the amount of toluene and remove the toluene and acetic anhydride by rotary evaporation under reduced pressure. Add an appropriate amount of dichloromethane to dissolve the formed dithiodiacetic anhydride, then add a dichloromethane solution of arachidyl alcohol and slowly add a dichloromethane solution of 4-dimethylaminopyridine (DMAP) dropwise. Stir magnetically at 25°C for 12 hours. The intermediate product, arachidyl alcohol-dithiodiacetic acid monoester, was isolated and purified by column chromatography using a cyclohexane-acetone elution system. The purified product from the previous step was added with a dichloromethane solution of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDCI), 1-hydroxybenzotriazole (HOBt), and 4-dimethylaminopyridine (DMAP). The mixture was activated in an ice bath at 0°C for 2 hours. A dichloromethane solution of cabazitaxel was then added and stirred at 25°C for 48 hours. After the reaction, the product was separated using a preparative phase to obtain a cabazitaxel-arachidyl alcohol prodrug with 2,2'-dithiodiacetic acid as the linker. During the reaction, the ratio of 2,2'-dithiodiacetic acid: acetic anhydride is 1:1, unit: mmol:ml; the molar ratio of DMAP: arachidyl alcohol: dithiodiacetic anhydride is 0.4:2:1, and the molar ratio of arachidyl alcohol-dithiodiacetic acid monoester: HOBt: EDCI: DMAP: cabazitaxel is 1:1:2:0.4:0.8.

[0118] Using mass spectrometry and 1 H-NMR confirmed the structure of the product. The spectral analysis results are as follows: 1H-NMR(600MHz,DMSO-d6)δ7.967(2H,t),7.662(3H,d),7.378-7.423(3H,m),7.153-7.189(2H,m),5.807(1H,d,-NH),5.355(1H,d,3'-H),5.124 (1H,d,2-H),5.065(4H,m,2'-H,-CH=CH-,5-H),4.924-4.954(1H,t,7-H ),4.685(1H,d,20α-H),4.477(1H,d,20β-H),3.993-4.012(2H,d,15α-H ,15β-H),3.861(3H,d,3-H),3.736(4H,m,6α-H,4-COCH3),2.647(1H,m,13-H),2.501(3H,t,14α-H,-CH2CO-),1.790(6H,s,-CH2CH=CHCH2-),1. 618(4H,s,18-H),1.503(t,5H,6β-H),1.377(10H,s,16-H,19-H,-CH2CH2CO-),1.240(24H,t,17-H),0.975(3H,t,-CH3),0.850(5H,t,-CH2CH3).

[0119] MS (ESI) m / z for C 69 H 101 NO 17 S2Na[M+Na] + :1280.

[0120] Example 6: Synthesis of Cabazitaxel-2-octyl-dodecanol Prodrug with 4,4'-dithiodibutyric Acid as a Linker

[0121] An appropriate amount of 4,4'-dithiodibutyric acid was dissolved in acetic anhydride and placed in a 25 mL eggplant-shaped flask. After complete dissolution, the mixture was magnetically stirred at 25°C for 2 hours and then transferred to a 100 mL eggplant-shaped flask. Three times the amount of toluene was added. The toluene and acetic anhydride were removed by rotary evaporation under reduced pressure. An appropriate amount of dichloromethane was added to dissolve the formed dithiodibutyric anhydride. Then, a dichloromethane solution of 2-octyl-dodecanol was added, and a dichloromethane solution of 4-dimethylaminopyridine (DMAP) was slowly added dropwise. The mixture was magnetically stirred at 25°C for 12 hours. The intermediate product, 2-octyl-dodecanol-dithiodibutyric acid monoester, was isolated and purified by column chromatography using a cyclohexane-acetone elution system. The purified product from the previous step was added with a dichloromethane solution of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDCI), 1-hydroxybenzotriazole (HOBt), and 4-dimethylaminopyridine (DMAP). The mixture was activated in an ice bath at 0°C for 2 hours, followed by the addition of a dichloromethane solution of cabazitaxel and stirred at 25°C for 48 hours. After the reaction, the product was separated using a preparative phase to obtain the cabazitaxel-2-octyl-dodecanol prodrug with 4,4'-dithiodibutyric acid as the linker. During the reaction, the ratio of 4,4'-dithiodibutyric acid: acetic anhydride is 1:1, unit: mmol:ml; the molar ratio of DMAP: 2-octyl-dodecanol: 4,4'-dithiodibutyric acid is 0.4:2:1, and the molar ratio of 2-octyl-dodecanol-dithiodibutyric acid monoester: HOBt: EDCI: DMAP: cabazitaxel is 1:1:2:0.4:0.8.

[0122] Using mass spectrometry and 1 H-NMR confirmed the structure of the product. The spectral analysis results are as follows: 1H-NMR(600MHz,DMSO-d6)δ7.965(2H,t),7.691(3H,d),7.349-7.425(3H,m),7.152-7.189(2H,m),5.753(1H,d,-NH),5.352(1H,d,3'-H),5.120( 1H,d,2-H),5.026-5.069(4H,m,2'-H,-CH=CH-,5-H),4.927-4.951(1H,t ,7-H),4.682(1H,d,20α-H),4.474(1H,d,20β-H),3.986-4.011(2H,d,15 α-H,15β-H),3.583(3H,d,3-H),3.495(4H,m,6α-H,4-COCH3),2.670(1H,m,13-H),2.501(3H,t,14α-H,-CH2CO-),1.780(6H,s,-CH2CH=CHCH2-),1 .609(4H,s,18-H),1.501(t,5H,6β-H),1.370(10H,s,16-H,19-H,-CH2CH 2CO-), 1.210(24H,t,17-H), 0.957(3H,t,-CH3), 0.831(5H,t,-CH2CH3).

[0123] MS (ESI) m / z for C 73 H 101 NO 17 S2Na[M+Na] + :1351.

[0124] Example 7: Synthesis of Cabazitaxel-2-octyl-dodecanol Prodrug with a Single Sulfide Bond as a Connecting Chain

[0125] An appropriate amount of monothiodiacetic acid was dissolved in acetic anhydride and placed in a 25 mL eggplant-shaped flask. After complete dissolution, the mixture was magnetically stirred at 25°C for 2 hours and then transferred to a 100 mL eggplant-shaped flask. Three times the amount of toluene was added. The toluene and acetic anhydride were removed by rotary evaporation under reduced pressure. An appropriate amount of dichloromethane was added to dissolve the monothiodiacetic anhydride formed. Then, a dichloromethane solution dissolved in 2-octyl-dodecanol was added, and a dichloromethane solution of 4-dimethylaminopyridine (DMAP) was slowly added dropwise. The mixture was magnetically stirred at 25°C for 12 hours to obtain the product. The intermediate product, 2-octyl-dodecanol monothiodiacetic acid monoester, was isolated and purified by column chromatography using a cyclohexane-acetone elution system. The purified product from the previous step was added with a dichloromethane solution of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDCI), 1-hydroxybenzotriazole (HOBt), and 4-dimethylaminopyridine (DMAP). The mixture was activated in an ice bath at 0°C for 2 hours, followed by the addition of a dichloromethane solution of cabazitaxel and the mixture was stirred at 25°C for 48 hours. After the reaction, the product was separated using a preparative phase to obtain a cabazitaxel-2-octyl-dodecanol prodrug with a monosulfide bond as the connecting linker. During the reaction, the ratio of monothiodiacetic acid: acetic anhydride is 1:1, unit: mmol:ml; the molar ratio of DMAP: 2-octyl-dodecanol: monothiodiacetic acid is 0.4:2:1, and the molar ratio of 2-octyl-dodecanol-monothiodiacetic acid monoester: HOBt: EDCI: DMAP: cabazitaxel is 1:1:2:0.4:0.8.

[0126] Using mass spectrometry and 1 H-NMR confirmed the structure of the product. The spectral analysis results are as follows: 1H-NMR(600MHz,DMSO-d6)δ7.965(2H,t),7.691(3H,d),7.349-7.425(3H,m),7.152-7.189(2H,m),5.753(1H,d,-NH),5.352(1H,d,3'-H),5.120( 1H,d,2-H),5.026-5.069(4H,m,2'-H,-CH=CH-,5-H),4.927-4.951(1H,t ,7-H),4.682(1H,d,20α-H),4.474(1H,d,20β-H),3.986-4.011(2H,d,15 α-H,15β-H),3.583(3H,d,3-H),3.495(4H,m,6α-H,4-COCH3),2.670(1H,m,13-H),2.501(3H,t,14α-H,-CH2CO-),1.780(6H,s,-CH2CH=CHCH2-),1 .609(4H,s,18-H),1.501(t,5H,6β-H),1.370(10H,s,16-H,19-H,-CH2CH 2CO-), 1.210(24H,t,17-H), 0.957(3H,t,-CH3), 0.831(5H,t,-CH2CH3).

[0127] MS (ESI) m / z for C 69 H 101 NO 17 SNa[M+Na] + :1271.

[0128] Example 8: Synthesis of Cabazitaxel-2-octyl-dodecanol Prodrug with a Single Selenium Bond as a Connecting Chain

[0129] An appropriate amount of monoselenodiacetic acid was dissolved in acetic anhydride and placed in a 25 mL eggplant-shaped flask. After complete dissolution, the mixture was magnetically stirred at 25°C for 2 hours and then transferred to a 100 mL eggplant-shaped flask. Three times the amount of toluene was added. The toluene and acetic anhydride were removed by rotary evaporation under reduced pressure. An appropriate amount of dichloromethane was added to dissolve the formed monoselenodiacetic anhydride. Then, a dichloromethane solution of 2-octyl-dodecanol was added, and a dichloromethane solution of 4-dimethylaminopyridine (DMAP) was slowly added dropwise. The mixture was magnetically stirred at 25°C for 12 hours to obtain the product. The intermediate product, 2-octyl-dodecanol-monoselenediacetic acid monoester, was isolated and purified by column chromatography using a cyclohexane-acetone elution system. The purified product from the previous step was added with a dichloromethane solution of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDCI), 1-hydroxybenzotriazole (HOBt), and 4-dimethylaminopyridine (DMAP). The mixture was activated in an ice bath at 0°C for 2 hours, followed by the addition of a dichloromethane solution of cabazitaxel and the mixture was stirred at 25°C for 48 hours. After the reaction, the product was separated using a preparative phase to obtain a cabazitaxel-2-octyl-dodecanol prodrug with a single selenium bond as the connecting linker. During the reaction, the ratio of monoselenodiacetic acid: acetic anhydride is 1:1, unit: mmol:ml; the molar ratio of DMAP: 2-octyl-dodecanol: monoselenodiacetic acid is 0.4:2:1, and the molar ratio of 2-octyl-dodecanol-monoselenediacetic acid monoester: HOBt: EDCI: DMAP: cabazitaxel is 1:1:2:0.4:0.8.

[0130] Using mass spectrometry and 1 H-NMR confirmed the structure of the product. The spectral analysis results are as follows: 1H-NMR(600MHz,DMSO-d6)δ7.969(2H,t),7.661(3H,d),7.385-7.428(3H,m),7.175(2H,m),5.807(1H,d,-NH),5.355(1H,d,3'-H),5. 117(1H,d,2-H),4.932-4.958(4H,m,2'-H,-CH=CH-,5-H),4.685(1H,d,20α-H),4.474(1H,d,20β-H),3.986-4.011(2H,d,15α-H,15β- H),3.571-3.829(3H,d,3-H)3.197(4H,m,6α-H,4-COCH3),2.670(1H,m,13-H),2.492(3H,t,14α-H,-CH2CO-),1.795(6H,s,-CH2CH=CH CH2-),1.501(4H,s,18-H),1.379(10H,s,16-H,19-H,-CH2CH2CO-),1.239(24H,t,17-H),0.958(3H,t,-CH3),0.850(5H,t,-CH2CH3).

[0131] MS (ESI) m / z for C 69 H 101 NO 17 SeNa[M+Na] + :1319.

[0132] Example 9: Preparation of PEG-modified small molecule prodrug self-assembled nanoparticles

[0133] Accurately weigh DSPE-PEG 2K 0.4 mg of the PEG-modified PEG-containing nanoparticles and 2 mg of the prodrug were dissolved in 200 μL of acetone. This ethanol solution was slowly added dropwise to 1.8 mL of deionized water with stirring, spontaneously forming uniformly sized PEG-modified nanoparticles. The acetone was removed by rotary evaporation under reduced pressure, yielding a nanocolloidal solution free of organic reagents. As shown in Table 1, with the exception of the CTX-SS-AA nanoparticles, the particle size of all the nanoparticles was approximately 100 nm, with a particle size distribution of less than 0.2 and a surface charge of approximately -20 mV.

[0134] Table 1. Particle size, size distribution and surface charge of PEG-modified prodrug self-assembled nanoparticles

[0135]

[0136] Results showed that cabazitaxel prodrugs with different linkers and fatty alcohol side chains could form self-assembled nanoparticles. Cabazitaxel-branched fatty alcohol small molecule prodrug self-assembled nanoparticles exhibited smaller particle size and size distribution than cabazitaxel-linear fatty alcohol small molecule prodrug self-assembled nanoparticles. The particle size of these cabazitaxel-branched fatty alcohol small molecule prodrug self-assembled nanoparticles was approximately 100 nm, with a very uniform size distribution of approximately 0.1. This facilitates tumor-targeted accumulation through the high permeability and retention of solid tumors. The surface charge of the nanoparticles was approximately -20 mV, which prevented aggregation through charge repulsion.

[0137] Example 10: Preparation of Lyophilized Powder of Small Molecule Prodrug Self-Assembled Nanoparticles

[0138] 1 mL of the PEGylated, single-sulfide-linked cabazitaxel-2-octyl-dodecanol prodrug self-assembled nanoparticles prepared in Example 9 was placed in a vial and lyoprotectants were added, respectively, to 5% sucrose, 10% sucrose, 5% lactose, 10% lactose, 5% trehalose, 10% trehalose, 5% mannitol, 10% mannitol, 5% glucose, and 10% glucose. The mixture was then freeze-dried in a lyophilizer for 24 hours to produce a white cake. The resulting lyophilized powder was reconstituted with deionized water and the particle size and surface charge were measured. The results are shown in Table 2. The results show that the prodrug self-assembled nanoparticles prepared using a single lyoprotectant exhibited a larger particle size and larger particle size distribution after reconstitution.

[0139] Table 2. Particle size, size distribution, and surface charge of lyophilized and reconstituted prodrug self-assembled nanoparticles

[0140]

[0141] Example 11: Preparation of small molecule prodrug self-assembled nanoparticle lyophilized powder

[0142] 1 mL of the PEGylated, single-sulfide-linked cabazitaxel-2-octyl-dodecanol prodrug self-assembled nanoparticles prepared in Example 9 was placed in a vial and lyoprotectants were added, respectively, with 5% lactose and 5% mannitol, 5% lactose and 5% maltose, 5% lactose and 5% sucrose, and 5% lactose and 5% trehalose. After the lyoprotectants were completely dissolved, the pellets were pre-frozen at -80°C for 12 hours and then freeze-dried in a freeze dryer for 24 hours to produce a white cake. The resulting lyophilized powder was reconstituted with deionized water and the particle size and surface charge were measured. The results are shown in Table 3. The results show that the lyophilized powder prepared with the combined protective agent exhibited a smaller particle size after reconstitution compared to the lyophilized powder prepared with a single protective agent.

[0143] Table 3. Particle size, size distribution, and surface charge of lyophilized and reconstituted prodrug self-assembled nanoparticles

[0144]

[0145] Example 12: Pharmacokinetic Study of Small Molecule Prodrug Self-Assembled Nanoparticles

[0146] 24 healthy male rats weighing 180-220 g were randomly divided into 4 groups. They were fasted for 12 hours before administration and had free access to water. The 4 groups were respectively injected with cabazitaxel solution and PEG-modified prodrug self-assembled nanoparticles prepared in Example 9 through the tail vein at a dose of 4 mg / kg (calculated as cabazitaxel). Blood was collected at the specified time points. The drug concentration in the blood was determined by liquid chromatography-mass spectrometry. The results are shown in Table 4. The results showed that the circulation time of cabazitaxel solution was very short and it was rapidly metabolized and cleared in the body after administration. In contrast, the circulation time of PEG-modified small molecule prodrug self-assembled nanoparticles was significantly prolonged, the bioavailability was significantly improved, and the pharmacokinetic parameters were significantly improved. The overall AUC of CTX-SS-OD nanoparticles, CTX-Se-OD nanoparticles and CTX-S-OD nanoparticles 0-24h (the sum of the prodrug and the parent drug) were 395.47 times, 277.37 times, and 260.19 times that of the cabazitaxel solution, respectively; 1 / 2 They were 19.72 times, 10.16 times, and 11.51 times that of cabazitaxel solution, respectively; C max The pharmacokinetic parameters of CTX-SS-OD nanoparticles were significantly improved, indicating that disulfide bonds have an advantage as connecting chains.

[0147] Table 4. Pharmacokinetic parameters of cabazitaxel solution and prodrug nanoparticles

[0148]

[0149] Example 13: In vivo anti-tumor experiment of PEGylated small molecule prodrug self-assembled nanoparticles

[0150] Mouse breast cancer cell suspension (4T1, 5x10 6 cells / 100 μL) were inoculated subcutaneously on the dorsal flank of female BALB / c mice. 3The tumor-bearing mice were randomly divided into 6 groups, with 8 mice in each group: normal saline group (Saline), cabazitaxel solution group, CTX-SS-HU nanoparticle group, CTX-SS-HD nanoparticle group, CTX-SS-OD nanoparticle group and CTX-SS-DT nanoparticle group. The nanoparticles used for administration were PEG-modified small molecule prodrug self-assembled nanoparticles prepared in Example 9, and the dosage was 10 mg / kg (calculated based on the cabazitaxel concentration). The drug was administered once every other day for 5 consecutive times. After administration, the survival status and weight changes of the mice were detected every day, and the tumor volume was measured. The mice were killed one day after the last administration, and the organs and tumors were obtained for further analysis and evaluation. The results are shown in the following table. Figure 1-3 shown. Figure 1 The results showed that in the saline group, the tumor volume increased rapidly, reaching 1000 mm on the 10th day. 3 In contrast, both nanoparticles and cabazitaxel solution significantly inhibited tumor growth (<200 mm 3 ); Figure 2 The results showed that the body weight of mice in the cabazitaxel solution group decreased significantly, while the body weight of mice in the prodrug self-assembled nanoparticle group did not change significantly; Figure 3 The results showed that all formulations caused a decrease in white blood cell counts in the mice, with the CTX-SS-OD and CTX-SS-HU nanoparticle groups causing the smallest decreases. These results demonstrate that self-assembled nanoparticles containing cabazitaxel and branched fatty alcohol small molecule prodrugs, chemically linked by disulfide bonds, exhibit potent anti-tumor effects without causing significant systemic toxicity, making them a safe and effective chemotherapy drug delivery system.

[0151] Example 14: In vivo anti-tumor experiment of PEGylated small molecule prodrug self-assembled nanoparticles

[0152] Mouse breast cancer cell suspension (4T1, 5x10 6 cells / 100 μL) were inoculated subcutaneously on the dorsal flank of female BALB / c mice. 3 The tumor-bearing mice were randomly divided into 7 groups, with 8 mice in each group: normal saline group (Saline), CTX-SS-OD nanoparticles 2mg / kg, 10mg / kg group, 20mg / kg group and CTX-SS-AA nanoparticles 2mg / kg, 10mg / kg group, 20mg / kg group. The nanoparticles used for administration were PEG-modified small molecule prodrug self-assembled nanoparticles prepared in Example 9, and the dosage was calculated based on the cabazitaxel concentration. The drug was administered once every other day for 5 consecutive times. After administration, the survival status and weight changes of the mice were detected every day, and the tumor volume was measured. The mice were killed one day after the last administration, and the organs and tumors were obtained for further analysis and evaluation. The results are as follows: Figure 4-8 shown. Figure 4 The results showed that in the saline group, the tumor volume increased rapidly, reaching 1000 mm on the 10th day. 3 The tumor volume in the CTX-SS-OD nanoparticle group was smaller than that in the CTX-SS-AA nanoparticle group at the same dose. Figure 5 It showed that when the administration dose was the same, the body weight of the CTX-SS-AA nanoparticle group decreased more seriously than that of the CTX-SS-OD nanoparticle group; Figure 6 The results showed that the prodrug self-assembled nanoparticles group had a lower tumor burden than the saline group. At the same concentration, the tumor burden of CTX-SS-OD nanoparticles was lower than that of CTX-SS-AA. Figure 7 It showed that when the administration dose was equivalent, the white blood cell count decreased less in the CTX-SS-OD nanoparticle group; Figure 8 The results showed that at a dose of 20 mg / kg, the urea nitrogen level in the CTX-SS-OD nanoparticle group increased less. The results showed that the self-assembled nanoparticles of cabazitaxel-branched fatty alcohol small molecule prodrug had a stronger anti-tumor effect than the self-assembled nanoparticles of cabazitaxel-linear fatty alcohol small molecule prodrug, while also having a better safety profile.

[0153] Example 15: In vivo anti-tumor experiment of PEGylated small molecule prodrug self-assembled nanoparticles

[0154] Mouse breast cancer cell suspension (4T1, 5x10 6 cells / 100 μL) were inoculated subcutaneously on the dorsal flank of female BALB / c mice. 3 The tumor-bearing mice were randomly divided into 5 groups, with 8 mice in each group: normal saline group (Saline), cabazitaxel solution group, CTX-S-OD nanoparticle group, CTX-SS-OD nanoparticle group, and CTX-Se-OD nanoparticle group. The nanoparticles used for administration were PEG-modified small molecule prodrug self-assembled nanoparticles prepared in Example 9, and the dosage was 4 mg / kg (calculated based on the cabazitaxel concentration). The drug was administered once every other day for 5 consecutive times. After administration, the survival status and weight changes of the mice were detected every day, and the tumor volume was measured. The mice were killed one day after the last administration, and the organs and tumors were obtained for further analysis and evaluation. The results are shown in the figure. Figure 9-11 shown. Figure 9 The results showed that in the saline group, the tumor volume increased rapidly, reaching 700 mm on the 10th day. 3 The tumor volumes of the CTX-S-OD nanoparticle group and the CTX-Se-OD nanoparticle group were similar (<400 mm 3 ), the tumor volumes of the CTX-SS-OD nanoparticle group and the cabazitaxel solution group were smaller (<200 mm3 ); Figure 10 The results showed that the body weight of the three nanoparticle groups did not change significantly, while the body weight of the cabazitaxel solution group decreased significantly; Figure 11 The results showed that there was no statistically significant difference in tumor burden between the CTX-SS-OD nanoparticle group and the cabazitaxel solution group. The results indicate that CTX-SS-OD nanoparticles not only have a good anti-tumor effect but also have a better safety profile than cabazitaxel solution. Furthermore, disulfide bonds are more advantageous as connecting chains than monosulfide bonds or monoselenium bonds.

[0155] Example 16: In vivo anti-tumor experiment of PEGylated small molecule prodrug self-assembled nanoparticles

[0156] Mouse breast cancer cell suspension (4T1, 5x10 6 cells / 100 μL) were inoculated subcutaneously on the dorsal flank of female BALB / c mice. 3 The tumor-bearing mice were randomly divided into 8 groups, with 8 mice in each group: normal saline group (Saline), cabazitaxel solution 2 mg / kg group, cabazitaxel solution 10 mg / kg group, cabazitaxel solution 15 mg / kg group, CTX-SS-OD nanoparticles 2 mg / kg group, CTX-SS-OD nanoparticles 10 mg / kg group, CTX-SS-OD nanoparticles 15 mg / kg group, CTX-SS-OD nanoparticles 20 mg / kg group. The nanoparticles used for administration were PEG-modified small molecule prodrug self-assembled nanoparticles prepared in Example 9, and the dosage was calculated based on the cabazitaxel concentration. The drug was administered once every other day for 5 consecutive times. The survival status and weight changes of the mice were detected every day, and the tumor volume was measured. The mice were killed one day after the last administration, and the organs and tumors were obtained for further analysis and evaluation. The results are as follows: Figure 12-14 shown. Figure 12 、 14 The results showed that in the blank control group, the tumor volume increased rapidly and reached 800 mm on the 10th day. 3 At a dose of 2 mg / kg, the tumor volume and tumor burden in the CTX-SS-OD nanoparticle group were lower than those in the cabazitaxel solution group. At doses of 10 mg / kg and 15 mg / kg, there were no statistically significant differences in tumor volume and tumor burden between the CTX-SS-OD nanoparticle group and the cabazitaxel solution group. Figure 13Results showed that at a 2 mg / kg dose, there was no significant change in body weight in the CTX-SS-OD nanoparticle group or the cabazitaxel solution group. At a 10 mg / kg dose, there was no change in body weight in the CTX-SS-OD nanoparticle group, while the cabazitaxel solution group experienced a decrease. At a 15 mg / kg dose, all mice in the cabazitaxel solution group died, while none in the CTX-SS-OD nanoparticle group died. These results suggest that CTX-SS-OD nanoparticles exhibit comparable antitumor efficacy to cabazitaxel solution while exhibiting lower toxicity.

[0157] Example 17: In vivo anti-tumor experiment of PEGylated small molecule prodrug self-assembled nanoparticles

[0158] Mouse breast cancer cell suspension (4T1, 5x10 6 cells / 100 μL) were inoculated subcutaneously on the dorsal flank of female BALB / c mice. 3 The tumor-bearing mice were randomly divided into 7 groups, with 8 mice in each group: normal saline group (Saline), CTX-SS-OD nanoparticles 20 mg / kg group, CTX-SS-OD nanoparticles 30 mg / kg group, CTX-SS-DT nanoparticles 20 mg / kg group, CTX-SS-DT nanoparticles 30 mg / kg group, γ-CTX-SS-OD nanoparticles 20 mg / kg group, γ-CTX-SS-OD nanoparticles 30 mg / kg group. The nanoparticles used for administration were PEG-modified small molecule prodrug self-assembled nanoparticles prepared in Example 9, and the dosage was calculated based on the cabazitaxel concentration. The drug was administered once every other day for 5 consecutive times. The survival status and weight changes of the mice were detected every day, and the tumor volume was measured. The mice were killed one day after the last administration, and the organs and tumors were obtained for further analysis and evaluation. The results are as follows: Figure 15-16 shown. Figure 15 The results showed that all three nanoparticles had good anti-tumor effects and significantly inhibited tumor growth; Figure 16 The results showed that when the dosage was equivalent, the γ-CTX-SS-OD group had the least weight loss, indicating that γ-CTX-SS-OD nanoparticles had better safety compared with CTX-SS-OD nanoparticles and CTX-SS-DT nanoparticles.

[0159] Example 18: Tolerance experiment of PEGylated small molecule prodrug self-assembled nanoparticles

[0160] Female BALB / c mice were divided into 11 groups of 3 mice each. Four of the groups received 30 mg / kg, 40 mg / kg, 50 mg / kg, and 60 mg / kg of cabazitaxel solution via tail vein injection. The other seven groups received 200 mg / kg of CTX-SS-HD nanoparticles, CTX-SS-HU nanoparticles, CTX-SS-OD nanoparticles, CTX-SS-DT nanoparticles, CTX-S-OD nanoparticles, CTX-SS-AA nanoparticles, and γ-CTX-SS-OD nanoparticles via tail vein injection every 8 hours until all mice died. The survival of the mice was observed after each dose, and the results are shown in Table 5. The results show that the prodrug self-assembled nanoparticles are much more tolerable than the cabazitaxel solution. Among the four nanoparticles containing different branched fatty alcohol side chains—CTX-SS-HD, CTX-SS-HU, CTX-SS-OD, and CTX-SS-DT—CTX-SS-OD nanoparticles demonstrated the best safety profile, indicating that 2-octyl-dodecanol as a side chain offers a safety advantage. Comparing side chains with different structures (branched or linear), CTX-SS-OD nanoparticles containing branched fatty alcohols demonstrated superior safety compared to CTX-SS-AA nanoparticles containing linear fatty alcohols, demonstrating that branched fatty alcohols as side chains offer a safety advantage over linear fatty alcohols. Comparing linker lengths, γ-CTX-SS-OD nanoparticles containing longer linkers demonstrated superior safety compared to CTX-SS-OD nanoparticles, indicating that 4,4'-dithiodibutyric acid as a linker offers a safety advantage. Among all nanoparticle formulations, γ-CTX-SS-OD nanoparticles demonstrated the highest maximum tolerated dose, 40 times that of cabazitaxel solution.

[0161] Table 5. Tolerated doses of cabazitaxel solution and prodrug nanoparticles

[0162]

Claims

1. Self-assembled nanoparticles of cabazitaxel-branched fatty alcohol small molecule prodrug, characterized in that: It is a PEG-modified prodrug self-assembled nanoparticle, wherein the cabazitaxel-fatty alcohol small molecule prodrug is selected from the following structural formula: Cabazitaxel-2-octyl-dodecanol prodrug CTX-SS-OD with 2,2'-dithiodiacetic acid as the linker: ； Cabazitaxel-2-octyl-dodecanol prodrug CTX-SS-OD with 2,2'-dithiodiacetic acid as the linker: ； The method for preparing the cabazitaxel-branched fatty alcohol small molecule prodrug self-assembled nanoparticles comprises the following steps: A PEG modifier and a cabazitaxel-branched fatty alcohol small molecule prodrug are dissolved in an organic solvent, and the solution is added dropwise to water under stirring, so that the prodrug spontaneously forms uniform nanoparticles. The organic solvent in the preparation is removed by a reduced pressure rotary evaporation method to obtain PEG-modified cabazitaxel-branched fatty alcohol small molecule prodrug self-assembled nanoparticles, wherein the mass ratio of the cabazitaxel-branched fatty alcohol small molecule prodrug to the PEG modifier is 1:(1-0.1), and the PEG modifier is DSPE-PEG.

2. Use of the self-assembled nanoparticles of cabazitaxel-branched fatty alcohol small molecule prodrug according to claim 1 in the preparation of anti-breast cancer drugs.

3. The lyophilized powder injection of self-assembled nanoparticles of cabazitaxel-branched fatty alcohol small molecule prodrug according to claim 1, characterized in that: The lyophilized powder injection comprises a cabazitaxel-branched fatty alcohol small molecule prodrug self-assembled nanoparticle solution and a lyoprotectant. The concentration of the self-assembled nanoparticle solution is 0.1 mg / mL-20 mg / mL. The lyoprotectant is one or more of glucose, galactose, trehalose, sucrose, mannitol, sorbitol, xylitol, polyethylene glycol, hydroxyethyl starch or dextran. The dosage of the lyoprotectant is 1%-20%, W / V.

Images