Preparation and use of albumin-paclitaxel twin drug nanoparticle

Abstract

A preparation method of an albumin-paclitaxel twin drug nanoparticle. The preparation particularly relates to mixing albumin with water to prepare an albumin solution; mixing paclitaxel or a paclitaxel twin drug compound with an organic solvent to obtain a drug solution; and mixing the albumin solution with the drug solution, and performing ultrasonic dispersion, and then rotary evaporation and ultrafiltration to prepare an albumin-binding paclitaxel or albumin-paclitaxel twin drug nanoparticle. Disclosed is a use of the albumin-paclitaxel twin drug nanoparticle. Drug accumulation in tumors can be increased by means of using the special GP60-caveolin-SPARC transport mechanism of albumin, and the anti-tumor effect is good.

Description

Preparation and application of a class of albumin-paclitaxel twin-drug nanoparticles Technical Field

[0001] The present invention belongs to the field of pharmaceutical chemistry and biopharmaceutical technology, and specifically relates to the preparation and application of a class of albumin-paclitaxel twin-drug nanoparticles. Background Art

[0002] Malignant tumors are a major global public health issue, with morbidity and mortality rates increasing annually, posing a serious threat to human life. Currently, chemotherapy remains the mainstay of treatment for malignant tumors, particularly pancreatic cancer. However, the clinical application of most anti-tumor drugs is severely limited by their low solubility, easy clearance by the body, and lack of tumor-specific targeting.

[0003] Albumin has excellent biocompatibility and water solubility, and its unique GP60-caveolin-SPARC transport mechanism enables tumor targeting and penetration of drugs, making it an ideal drug delivery vehicle. Furthermore, simple physical methods such as ultrasound and stirring can be used to prepare it into uniform nanoparticles, which enhance drug accumulation in tumor tissues through the EPR effect, achieving enhanced efficacy and reduced toxicity of the drug. In 2005, Abraxane (an albumin-bound paclitaxel nanocomplex) developed by Celgene Corporation in the United States was approved for marketing by the FDA. It has now become a major chemotherapy drug for a variety of malignant tumors, including metastatic breast cancer, non-small cell lung cancer, and pancreatic cancer.

[0004] Since tumors often have metabolic abnormalities and complex microenvironments, the efficacy of chemotherapy drugs alone is often poor. The development of multi-drug combination therapy is expected to bring new hope to patients. At present, the combination of Abraxane with anti-tumor drugs such as carboplatin and gemcitabine has become the first choice for first-line treatment of various cancers due to its good therapeutic effect. Although combination therapy can significantly improve the survival rate of patients, there are still many problems that need to be solved, such as low bioavailability of free drugs such as carboplatin and gemcitabine, poor targeting, and obvious toxic side effects. In addition, the most difficult part of combination therapy is the large differences in the pharmacokinetics and pharmacodynamics of different drugs, unclear in vivo efficacy, and complex medication regimens. These factors have severely limited the development of multi-drug combination therapy. Summary of the Invention

[0005] In response to the above problems, we used paclitaxel as the target head and covalently linked it with cytotoxic drugs, small molecule targeted drugs, immune checkpoint inhibitors, hormone drugs and immune agonists to construct a class of paclitaxel twin drugs. We then reacted with albumin to form a uniform albumin-paclitaxel twin drug nanoparticle complex, thereby achieving efficient co-loading of multiple drugs and releasing the drug molecules in the tumor area through targeted delivery, achieving the purpose of treating and regressing the tumor. This technology is expected to break through the current efficacy bottleneck of malignant tumors.

[0006] The purpose of the present invention is to provide a preparation and application of albumin-paclitaxel twin-drug nanoparticles that have anti-proliferation, promote killing, prevent cancer cell metastasis and have good stability.

[0007] The first purpose of the present invention is to provide a method for synthesizing paclitaxel twin drugs by coupling paclitaxel to anti-tumor drugs through different connecting bonds, thereby achieving efficient co-loading of multiple drugs and specific response release of the loaded drugs in tumor cells. The anti-tumor drugs include but are not limited to cytotoxic drugs, small molecule targeted drugs, immune checkpoint inhibitors, hormone drugs and immune agonists.

[0008] The second purpose of the present invention is to construct a series of uniform and stable albumin-paclitaxel twin-drug nanocomplexes using albumin encapsulation technology, wherein the nanocomplexes include but are not limited to single-drug, dual-drug and multi-drug forms.

[0009] The third object of the present invention is to provide and compare the anti-tumor effects of the above-mentioned albumin-paclitaxel twin-drug nanocomplex.

[0010] The paclitaxel twin drug of the present invention is paclitaxel and an anti-tumor drug connected by a covalent chemical bond.

[0011] In one aspect, the present invention provides a paclitaxel twin-drug compound having the following structural formula:

[0012] The R is a drug molecule, including any one of a cytotoxic drug, a small molecule targeted drug, an immune checkpoint inhibitor, a hormone drug, and an immune agonist; the structure of the Linker is any one of the following: wherein n is any integer from 1 to 14, m is any integer from 1 to 14, R1 is at least one of a hydrazone bond, a hydrazide bond, a disulfide bond, a thioether bond, a diselenide bond, a selenoether bond, a thioketal bond, and a Michael adduct; R2 is at least one of a hydrazone bond, a hydrazide bond, a disulfide bond, a thioether bond, a diselenide bond, a selenoether bond, a thioketal bond, and a Michael adduct; R3 is at least one of a hydrazone bond, a hydrazide bond, a disulfide bond, a thioether bond, a diselenide bond, a selenoether bond, a thioketal bond, and a Michael adduct; and R4 is at least one of a hydrazone bond, a hydrazide bond, a disulfide bond, a thioether bond, a diselenide bond, a selenoether bond, a thioketal bond, and a Michael adduct.

[0013] Furthermore, the drug molecule is any one of T785, di-ABZI, SR07, Crizotinb, Palbociclib, MK1775, MK2206, AZD7762, Ceritinib, Navitoclax, Dasatinib and Exatecan; the structure of the linker is shown as follows:

[0014] Preferably, the drug molecule is any one of Linsitinib, MK2206, Navitoclax, and Dasatinib.

[0015] In some embodiments, the present invention demonstrates through CCK8 experiments that paclitaxel, when used in combination with Crizotinb, Dasatinib, Linsitinib, MK2206, and Exatecan drug molecules, can significantly enhance the inhibitory effect of the paclitaxel twin-drug compound on tumor cell proliferation; when used in combination with Navitoclax, the inhibitory effect of the drug molecule on PATU-8988T cell proliferation is significantly enhanced; at the same time, the synergistic index of paclitaxel when used in combination with Linsitinib, Navitoclax, MK2206, and AZD7762 drug molecules is significantly greater than 1, and the synergistic index of paclitaxel when used in combination with Dasatinib for inhibiting PATU-8988T cell proliferation is also significantly greater than 1, indicating that paclitaxel has a significant synergistic effect in inhibiting tumor proliferation when used in combination with Linsitinib, Navitoclax, MK2206, and Dasatinib drug molecules.

[0016] On the other hand, the present invention provides a method for preparing the above-mentioned paclitaxel twin-drug compound, comprising: preparing the paclitaxel twin-drug compound by reacting the drug molecule, linker and paclitaxel.

[0017] Furthermore, the linker molecule reacts with NpCl in a solvent to produce the intermediate SS-2, which is then reacted with PTX in dichloromethane to produce PTX-SS-Np. Finally, PTX-SS-Np reacts with the drug molecule in DMF to produce the paclitaxel twin-drug compound. The linker molecule is SS-1.

[0018] Preferably, a solvent is used in the chemical reaction, and the solvent is one or more of dichloromethane, chloroform, 1,2-dichloroethane, tetrahydrofuran, acetonitrile, N,N-dimethylformamide, 1,4-dioxane, dimethyl sulfoxide, benzene, toluene, xylene, chlorobenzene, and o-dichlorobenzene; or, an alkaline reagent is used in the chemical reaction, and the alkaline reagent is one or more of alkali metal hydroxide, alkaline earth metal hydroxide, alkali metal or alkaline earth metal carbonate, alkali metal or alkaline earth metal bicarbonate, triethylamine, tributylamine, trioctylamine, N,N-diisopropylethylamine, pyridine, 4-dimethylaminopyridine, piperidine, N-methylmorpholine, N-methylpiperidine, tetrahydropyrrole, triethylenediamine, and tetrabutylammonium hydroxide; or, a condensing agent is used in the chemical reaction, and the condensing agent is one or more of EDCI, DCC, and DIC.

[0019] In another aspect, the present invention discloses a paclitaxel twin-drug nanoparticle comprising the above-mentioned paclitaxel twin-drug compound.

[0020] Furthermore, the paclitaxel twin-drug nanoparticles also include albumin, and the paclitaxel twin-drug compound is bonded to the albumin.

[0021] More preferably, the albumin includes at least one of bovine serum albumin, human serum albumin, ovalbumin and recombinant human serum albumin; the bonding is a non-covalent bond; and the paclitaxel twin-drug nanoparticles include at least one paclitaxel twin-drug compound.

[0022] Preferably, the albumin comprises human serum albumin.

[0023] Preferably, the paclitaxel twin-drug nanoparticles include 1 paclitaxel twin-drug compound; or, the paclitaxel twin-drug nanoparticles include 2 paclitaxel twin-drug compounds; or, the paclitaxel twin-drug nanoparticles include 3 paclitaxel twin-drug compounds; or, the paclitaxel twin-drug nanoparticles include 4 paclitaxel twin-drug compounds; or, the paclitaxel twin-drug nanoparticles include 5 paclitaxel twin-drug compounds.

[0024] Preferably, the paclitaxel twin-drug compound includes any one drug molecule selected from Linsitinib, MK2206, Navitoclax, and Dasatinib.

[0025] Furthermore, the hydrated particle size of the paclitaxel twin-drug nanoparticles is 90-140 nm, the surface potential is negative, and the PDI is less than 0.24.

[0026] Preferably, the hydrated particle size of the paclitaxel twin-drug nanoparticles is 90-120 nm.

[0027] In some embodiments, the present invention measured the particle size and distribution, PDI, and surface potential of paclitaxel twin-drug nanoparticles, including single-drug, dual-drug, and triple-drug formulations. The results revealed significant variation in particle size among different paclitaxel twin-drug nanoparticles. Nanoparticles Nab-PTX-SS-Linsitinib and Nab-PTX-SS-Navitoclax, with hydrated particle sizes less than 100 nm, exhibited the best tumor growth inhibition effects. Nanoparticles Nab-PTX-SS-Dasatinib and Nab-PTX-SS-MK2206, with hydrated particle sizes less than 120 nm, also exhibited significant anti-tumor activity. However, nanoparticles with larger particle sizes, such as Nab-PTX-SS-Pal@Cer@SR07, exhibited less potent anti-tumor activity than these nanoparticles. This may be due to the smaller particle size enhancing the nanoparticle's accumulation and distribution within tumor cells.

[0028] The present invention demonstrates through experiments that albumin-paclitaxel twin-drug nanoparticles comprising one, two, or three paclitaxel twin-drug compounds can significantly inhibit tumor growth. Specifically, when the drug molecules of the paclitaxel twin-drug compound particles in the albumin-paclitaxel twin-drug nanoparticles have a synergistic effect with paclitaxel on inhibiting tumor proliferation, and when the nanoparticles have a small particle size, they can significantly inhibit tumor growth. For example, the nanoparticles Nab-PTX-SS-Linsitinib, Nab-PTX-SS-Navitoclax, Nab-PTX-SS-Dasatinib, and Nab-PTX-SS-MK2206 can significantly inhibit tumor growth, resulting in minimal changes in tumor volume growth, and have better anti-tumor effects than other single-drug, dual-drug, or triple-drug nanoparticles.

[0029] In another aspect, the present invention discloses a method for preparing paclitaxel twin-drug nanoparticles, comprising: mixing the above-mentioned paclitaxel twin-drug compound and albumin in a composite liquid to prepare paclitaxel twin-drug nanoparticles.

[0030] Preferably, the paclitaxel twin-drug compound is prepared into a drug solution in an organic solvent; the drug solution is mixed with an albumin solution to prepare a nanosuspension; the organic solvent in the nanosuspension is evaporated and concentrated by ultrafiltration to obtain paclitaxel twin-drug nanoparticles.

[0031] Preferably, the organic solvent includes one or more of dichloromethane, chloroform, ethanol, DMSO, DMF and acetonitrile.

[0032] Preferably, the albumin solution is prepared by mixing albumin and water, and the concentration of albumin in the albumin solution is 1-100 mg / mL.

[0033] Preferably, an ultrasonic instrument is used in the nanosuspension, and the ultrasonic instrument includes but is not limited to an ultrasonic crusher, an ultrasonic cleaner, and the like.

[0034] In another aspect, the present invention discloses the use of the above-mentioned paclitaxel twin-drug compound in the preparation of a drug for treating cancer, including but not limited to non-small cell lung cancer, pancreatic cancer, cervical cancer, ovarian cancer, bladder cancer, breast cancer, lung cancer, gastric cancer and melanoma.

[0035] Furthermore, the paclitaxel twin-drug compound includes any one drug molecule selected from T785, di-ABZI, SR07, Crizotinb, Palbociclib, MK1775, MK2206, AZD7762, Ceritinib, Navitoclax, Dasatinib and Exatecan.

[0036] In another aspect, the present invention discloses the use of the above-mentioned paclitaxel twin-drug nanoparticles in the preparation of drugs for treating cancer, including but not limited to non-small cell lung cancer, pancreatic cancer, cervical cancer, ovarian cancer, bladder cancer, breast cancer, lung cancer, gastric cancer and melanoma.

[0037] Furthermore, the paclitaxel twin-drug nanoparticles also include albumin, and the paclitaxel twin-drug compound is bonded to the albumin; the albumin includes at least one of bovine serum albumin, human serum albumin, ovalbumin and recombinant human serum albumin; the bonding is a non-covalent bond; the paclitaxel twin-drug nanoparticles include at least one paclitaxel twin-drug compound.

[0038] The preparation route of the paclitaxel twin-drug compound is as follows:

[0039] Preferably, in the preparation of intermediate SS-2, DMAP is added to dichloromethane, NpCl and SS-1 are added under ice bath, and the mixture is stirred at 20-40° C. for 12-48 h. The reaction is monitored by TLC. After completion of the reaction, column chromatography is performed to obtain SS-2.

[0040] More preferably, in the preparation of intermediate SS-2, the usage ratio of DMAP to dichloromethane is 2-8 g / mL.

[0041] More preferably, in the preparation of intermediate SS-2, the ratio of NpCl to dichloromethane is 5-20 g / mL.

[0042] More preferably, in the preparation of intermediate SS-2, the amount of SS-1 used is 20-30 wt % of NpCl.

[0043] Preferably, in the preparation of the intermediate PTX-SS-Np, SS-2 is added to dichloromethane, and then PTX and DIEA are added, and stirred at 30-50° C. for 5-24 h. The reaction is monitored by TLC. After the reaction is completed, column chromatography is performed to obtain PTX-SS-Np.

[0044] More preferably, in the preparation of the intermediate PTX-SS-Np, the usage ratio of SS-2 to dichloromethane is 0.1-1 g / mL.

[0045] More preferably, in the preparation of the intermediate PTX-SS-Np, the amount of PTX used is 40-80 wt % of SS-2.

[0046] More preferably, in the preparation of the intermediate PTX-SS-Np, the usage ratio of DIEA and SS-2 is 0.1-0.5 mL / g.

[0047] Preferably, in the preparation of the paclitaxel twin-drug compound, PTX-SS-Np is added to DMF, and then the drug molecule and DIEA are added and mixed, and stirred at 30-50° C. for 5-24 hours. The reaction is monitored by TLC. After the reaction is completed, column chromatography is performed to separate the paclitaxel twin-drug compound.

[0048] More preferably, in the preparation of the paclitaxel twin-drug compound, the usage ratio of PTX-SS-Np and DMF is 0.05-0.5 g / mL.

[0049] More preferably, in the preparation of the paclitaxel twin-drug compound, the amount of the drug molecule used is 20-140 wt % of PTX-SS-Np.

[0050] More preferably, in the preparation of the paclitaxel twin-drug compound, the usage ratio of DIEA and PTX-SS-Np is 0.2-1 mL / g.

[0051] Preferably, in the preparation of paclitaxel twin-drug nanoparticles, the paclitaxel twin-drug compound is added to chloroform, and then ethanol is added to prepare a drug solution, the drug solution is mixed with an albumin solution to prepare a nanosuspension, the organic solvent in the nanosuspension is evaporated to remove, and ultrafiltration and concentration are performed to obtain paclitaxel twin-drug nanoparticles.

[0052] More preferably, in the preparation of paclitaxel twin-drug nanoparticles, the usage ratio of paclitaxel twin-drug compound and chloroform is 0.3-5 mg / μL.

[0053] More preferably, in the preparation of paclitaxel twin-drug nanoparticles, the volume of ethanol used is 5-20% of the volume of chloroform used.

[0054] More preferably, in the preparation of paclitaxel twin-drug nanoparticles, the albumin solution is prepared by mixing albumin and water, and the concentration of albumin in the albumin solution is 1-100 mg / mL.

[0055] More preferably, in the preparation of paclitaxel twin-drug nanoparticles, when the drug solution and the albumin solution are mixed, the amount of the drug solution used is based on the amount of chloroform used therein, and the volume of chloroform used is 1-10% of the volume of the albumin solution used.

[0056] In another aspect, the present invention discloses a method for preparing albumin-paclitaxel twin-drug nanoparticles, characterized in that it comprises the following steps:

[0057] S1, dissolving the paclitaxel twin-drug compound in a mixture of organic solvents to prepare a drug solution;

[0058] S2, preparing an albumin aqueous solution as the albumin solution;

[0059] S3, adding the drug solution to the albumin solution and preparing the nanosuspension by ultrasound;

[0060] S4, evaporating and removing the residual organic solvent in the suspension obtained in step S3;

[0061] S5, concentrating the nanosuspension by ultrafiltration to obtain albumin-paclitaxel twin-drug nanoparticles.

[0062] Preferably, the organic solvent in step S1 comprises one or more of dichloromethane, chloroform, ethanol, DMSO, DMF, and acetonitrile; the concentration of albumin in the albumin solution in step S2 is 1-100 mg / mL; and the ultrasonic instrument used in step S3 includes but is not limited to an ultrasonic disruptor and an ultrasonic cleaner.

[0063] When preparing albumin-paclitaxel twin-drug nanoparticles of the present invention, drug solutions of two different drugs can be selected and added to the albumin solution, and a nanosuspension is prepared by ultrasound, and then the albumin-paclitaxel twin-drug nanoparticles containing the two drugs are prepared through subsequent steps.

[0064] When preparing albumin-paclitaxel twin-drug nanoparticles of the present invention, drug solutions of three different drugs can be selected and added to the albumin solution, and a nanosuspension is prepared by ultrasound, and then the albumin-paclitaxel twin-drug nanoparticles containing the three drugs are prepared through subsequent steps.

[0065] After the paclitaxel twin-drug nanoparticles or albumin-paclitaxel twin-drug nanoparticles are prepared in the present invention, the obtained nanoparticles can be configured for intravenous administration, oral administration, subcutaneous injection or direct injection into the tumor / near the cancer.

[0066] In another aspect, the present invention discloses the preparation and application of albumin-bound paclitaxel nanoparticles (Nab-PTX).

[0067] In another aspect, the present invention discloses the preparation and application of albumin-paclitaxel derived twin-drug nanoparticles (Nab-pDCs).

[0068] The present invention has the following beneficial effects:

[0069] Based on paclitaxel, the present invention designs a class of paclitaxel twin drugs that respond to tumor tissue microenvironment stimulation, which can achieve responsive release of anti-tumor drugs, reduce the toxic side effects of drugs on non-tumor cells, and thus improve the therapeutic effect.

[0070] The present invention provides a series of preparation methods for paclitaxel twin-drug nanoparticles with albumin (such as human serum albumin) as an encapsulation carrier, comprising: preferably mixing human serum albumin with water to prepare an albumin solution; mixing paclitaxel or a paclitaxel twin-drug with an organic solvent to obtain a drug solution; mixing the albumin solution with the drug solution, and then ultrasonically dispersing and performing rotary evaporation and ultrafiltration to obtain albumin-bound paclitaxel or albumin-paclitaxel twin-drug nanoparticles.

[0071] In the technical solution disclosed in the present invention, the nanoparticle aqueous solution is stable at room temperature for at least 2 months or longer.

[0072] The drug-containing nanoparticles prepared by the present invention have anti-tumor effects, including but not limited to anti-proliferation, promoting killing, preventing cancer cell metastasis, etc.

[0073] The present invention utilizes methods such as mixing albumin with water to produce an albumin solution; mixing paclitaxel or a paclitaxel twin-drug compound with an organic solvent to produce a drug solution; and mixing the albumin solution with the drug solution, followed by ultrasonic dispersion, rotary evaporation, and ultrafiltration to produce albumin-bound paclitaxel or albumin-paclitaxel twin-drug nanoparticles. The prepared albumin-paclitaxel twin-drug nanoparticles utilize albumin's unique GP60-caveolin-SPARC transport mechanism to increase drug accumulation in tumors. Both in vitro and in vivo experiments demonstrate that the albumin-paclitaxel twin-drug nanoparticles possess excellent anti-tumor effects. Therefore, the present invention provides a method for preparing and applying albumin-paclitaxel twin-drug nanoparticles that exhibit anti-proliferation, promote killing, prevent cancer cell metastasis, and exhibit excellent stability. BRIEF DESCRIPTION OF THE DRAWINGS

[0074] FIG1 is a diagram showing the particle size and surface potential of albumin-paclitaxel twin-drug nanoparticles;

[0075] FIG2 is a diagram showing the cytotoxicity and combination index of paclitaxel in combination with anti-tumor drugs;

[0076] FIG3 is a graph showing the changes in tumor volume in pancreatic cancer-bearing C57 mice after intravenous administration;

[0077] FIG4 is a graph showing changes in body weight of pancreatic cancer-bearing C57 mice after intravenous administration. DETAILED DESCRIPTION

[0078] The technical solution of the present invention is further described in detail below in conjunction with the specific embodiments and the accompanying drawings: The synthesis route of the paclitaxel twin-drug compound is as follows:

[0079] Example 1: Synthesis of intermediate SS-2

[0080] Weigh DMAP (3.6 g, 29.47 mmol) and dissolve it in ultra-dry CH2Cl2 (100 mL). Slowly add NpCl (12.0 g, 59.53 mmol) and SS-1 (3.0 g, 19.48 mmol) under ice bath and stir at room temperature. After 24 hours, the reaction was monitored by TLC. After the reaction was complete, SS-2 (5.3 g, 56%) was obtained by column chromatography. 1 HNMR (400MHz, CDCl) δ (ppm): 8.24 (d, J = 10Hz, 4H), 7.36 (d, J = 10Hz, 4H), 4.55 (t, J = 4Hz, 4H), 3.08 (t, J = 4Hz, 4H).

[0081] Example 2: Synthesis of intermediate PTX-SS-Np

[0082] SS-2 (4.0 g, 8.26 mmol) was weighed and dissolved in ultra-dry CH2Cl2 (10 mL). PTX (2.3 g, 2.69 mmol) and N,N-diisopropylethylamine (DIEA, 1.0 mL) were added and stirred at 40°C (900 rpm) for 10 hours. The reaction was monitored by TLC. After completion, column chromatography was used to separate PTX-SS-Np (2.1 g, 65.6%). 1HNMR (400MHz, CDCl3) δ (ppm): 8.32-8.26 (m, 2H), 8.16-8.10 (d, J = 8, 2H), 7.66-7.59 (m, 1H), 7.54-7.48 (m2H), 7.45-7.38 (m, 4H), 7. 36-7.31(m,3H),6.34-6.27(br,1H),5.67-5.65(d,J=4,1H),5.55-5.46(m,2H),5.29(s,1H),5.03-4.99(d,J=8,1H),4.85(s,1H),4 .59-4.47(m,2H),4.47-4.31(m,3H),4.21-4.16(m,1H),3.95-3.84(m,2H),3.47(s,3H),3.33(s,3H),3.06-2.91(m,4H),2.77-2.67 (m,1H),2.47(s,3H),2.35-2.20(m,2H),2.03(s,3H),1.85-1.77(m,1H),1.75(s,3H),1.36(s,9H),1.27(s,1H),1.25-1.21(m,6H).

[0083] Example 3: Synthesis of PTX-SS-T785

[0084] PTX-SS-Np (200 mg, 0.17 mmol) was weighed and dissolved in ultra-dry DMF (2 mL), and compound T785 (0.2 mmol, 62.2 mg) and ultra-dry DIEA (100 μL) were added, mixed well, and stirred at 40 °C. The reaction was monitored by TLC for 10 hours. After completion of the reaction, PTX-SS-T785 (165 mg, 72%) was obtained by column chromatography. 1HNMR (600MHz, DMSO-d) δ (ppm): 9.22 (d, J = 8.5Hz, 1H), 8.42 (d, J = 9.6Hz, 1H), 8.30 (s, 0H), 8 .12–8.05(m,3H),8.02–7.94(m,8H),7.89–7.81(m,2H),7.78–7.67(m,3H),7.68–7.51(m,1 1H),7.51–7.38(m,15H),7.29–7.11(m,7H),6.42(s,5H),6.30(d,J=1.7Hz,2H),5.86(dd,J =20.3,10.0Hz,2H),5.58(t,J=8.6Hz,1H),5.47–5.24(m,6H),5.06–4.74(m,4H),4.60(d,J =10.6Hz,2H),4.49(q,J=7.3Hz,5H),4.37(td,J=6.1,3.0Hz,2H),4.22(t,J=6.5Hz,2H),4. 12(q,J=6.2Hz,6H),4.08–4.00(m,5H),3.62(dd,J=14.3,7.2Hz,3H),3.11–2.96(m,7H),2. 94–2.81 (m, 9H), 2.41–2.30 (m, 1H), 2.27 (d, J = 7.4 Hz, 6H), 2.10 (d, J = 1.3 Hz, 6H), 1.84–1.69 (m, 6H), 1.73–1.39 (m, 19H), 1.24 (s, 6H), 1.11–0.99 (m, 14H), 0.95 (td, J = 7.4, 3.8 Hz, 6H). HR-MS (ESI): [PTX-SS-T785+H]+ requires: 1371.5200, found m / z = 1371.5141.

[0085] Example 4: Synthesis of PTX-SS-diABZI

[0086] PTX-SS-Np (100 mg, 0.083 mmol) was weighed and dissolved in ultra-dry DMF (2 mL). Compound di-ABZI (0.1 mmol, 77.9 mg) and ultra-dry DIEA (100 μL) were then added and mixed evenly. The mixture was stirred (900 rpm) at 40°C for 10 hours and the reaction was monitored by TLC. After the reaction was completed, PTX-SS-diABZI (60 mg, 39%) was separated by column chromatography. 1HNMR(400MHz,Chloroform-d)δ8.17–8.03(m,2H),8.00(d,J=7.7Hz,1H),7.85(d,J=7.8Hz,1H),7.68(d,J=7.6Hz,2H),7.60–7 .22(m,16H),6.81(d,J=35.0Hz,4H),6.71–6.54(m,2H),6.22(dd,J=26.6,15.3Hz,4H),5.93(dd,J=9.1,3.7Hz,1H),5.74–5.2 5(m,8H),5.06–4.73(m,6H),4.58(d,J=8.9Hz,6H),4.45–3.94(m,12H),3.86–3.61(m,4H),3.20(s,12H),3.08–2.56(m,6H),2 .58–2.05(m,12H),2.02–1.50(m,12H),1.47–1.30(m,8H),1.31–1.11(m,16H),1.08(d,J=11.6Hz,4H),0.81(t,J=7.0Hz,3H). HR-MS(ESI):[PTX-SS-diABZI+H] + Theoretical value: 1839.6706, measured value m / z = 1839.6501.

[0087] Example 5: Synthesis of PTX-SS-SR07

[0088] PTX-SS-Np (200 mg, 0.17 mmol) was dissolved in ultra-dry DMF (2 mL). Compound SR07 (0.2 mmol, 77.6 mg) and ultra-dry DIEA (100 μL) were added, mixed thoroughly, and stirred at 40°C (900 rpm) for 10 hours. The reaction was monitored by TLC. After completion, PTX-SS-SR07 (145 mg, 60%) was isolated by column chromatography. 1HNMR(600MHz,DMSO-d)δ13.00(s,1H),9.22(d,J=8.5Hz,1H),9.12(d,J=8.7Hz,0H),8.89–8.7 8(m,2H),8.55(dd,J=9.1,1.3Hz,1H),8.46(d,J=9.1Hz,1H),8.26–8.18(m,2H),8.10–7.93(m ,2H),7.89–7.81(m,2H),7.78–7.68(m,1H),7.67–7.61(m,2H),7.62–7.51(m,2H),7.51–7.39 (m,5H),7.32–7.25(m,1H),7.21(ddt,J=6.4,3.8,2.0Hz,1H),6.30(t,J=3.9Hz,1H),5.94–5. 80(m,1H),5.57(td,J=8.6,1.4Hz,1H),5.47–5.29(m,2H),5.01–4.79(m,2H),4.68–4.56(m,1 H),4.37(t,J=5.3Hz,3H),4.33–3.92(m,6H),3.60(q,J=6.8,6.2Hz,1H),3.45(q,J=5.4Hz,2H ),3.07–2.82(m,3H),2.71–2.56(m,1H),2.45–2.17(m,4H),2.10(t,J=2.1Hz,3H),1.94–1.74 (m,4H),1.70–1.53(m,2H),1.50(d,J=3.2Hz,3H),1.24(s,2H),1.02(dd,J=14.1,3.5Hz,6H). HR-MS(ESI):[PTX-SS-SR07+H] + Theoretical value: 1452.4135, measured value m / z = 1452.4234.

[0089] Example 6: Synthesis of PTX-SS-Cri

[0090] PTX-SS-Np (200 mg, 0.17 mmol) was dissolved in ultra-dry DMF (2 mL). Crizotinb (0.2 mmol, 89.8 mg) and ultra-dry DIEA (100 μL) were added, mixed thoroughly, and stirred at 900 rpm at 40°C for 10 hours. The reaction was monitored by TLC. After completion, PTX-SS-Crizotinb (228 mg, 90%) was isolated by column chromatography. 1HNMR(600MHz,DMSO-d6)δ9.23(d,J=8.5Hz,1H),8.44(d,J=9.5Hz,1H),8.12–8.06(m,2H),8. 03–7.91(m,6H),7.88–7.83(m,2H),7.77–7.68(m,5H),7.65(t,J=7.7Hz,2H),7.63–7.52(m,1 0H),7.52–7.38(m,13H),7.22(tt,J=6.5,1.7Hz,2H),6.92(t,J=2.0Hz,2H),6.31(d,J=2.0Hz ,2H),6.09(qd,J=6.7,2.1Hz,2H),5.93–5.82(m,2H),5.68(s,4H),5.59(t,J=8.6Hz,1H),5.5 1–5.30(m,5H),4.96–4.74(m,4H),4.61(d,J=11.6Hz,2H),4.44–4.18(m,8H),4.17–3.93(m, 10H),3.70–3.57(m,3H),3.11–2.92(m,9H),2.90(s,1H),2.81(t,J=6.6Hz,1H),2.74(s,1H), 2.43–2.26(m,7H),2.10(d,J=3.0Hz,6H),2.05–1.97(m,5H),1.91–1.78(m,14H),1.75–1.63( m,2H),1.58(dd,J=15.3,8.9Hz,1H),1.52(d,J=1.7Hz,6H),1.25(s,6H),1.13–0.94(m,12H). HR-MS(ESI):[PTX-SS-Cri+H] + Theoretical value: 1509.4275, measured value m / z = 1509.4279.

[0091] Example 7: Synthesis of PTX-SS-Palb

[0092] PTX-SS-Np (200 mg, 0.17 mmol) was dissolved in ultra-dry DMF (2 mL). Palbociclib (0.2 mmol, 89.5 mg) and ultra-dry DIEA (100 μL) were added, mixed thoroughly, and stirred at 40°C (900 rpm) for 10 hours. The reaction was monitored by TLC. After completion, PTX-SS-Palb (208 mg, 83%) was isolated by column chromatography. 1HNMR(600MHz,DMSO-d6)δ10.02(s,1H),9.23(d,J=8.5Hz,1H),8.96(d,J=1.3Hz,1H),8.13–8.0 4(m,1H),8.03–7.94(m,2H),7.92–7.84(m,3H),7.78–7.70(m,1H),7.69–7.62(m,2H),7.62–7. 53(m,1H),7.52–7.41(m,7H),7.22(tt,J=6.2,2.3Hz,1H),6.31(s,1H),5.97–5.76(m,2H),5.5 9(t,J=8.6Hz,1H),5.44(d,J=7.1Hz,1H),5.40(d,J=8.7Hz,1H),4.92(dd,J=9.6,2.2Hz,1H),4 .84(d,J=7.1Hz,1H),4.60(s,1H),4.48–4.35(m,2H),4.25(t,J=6.1Hz,2H),4.13(dt,J=11.1, 7.0Hz,1H),4.07–3.99(m,2H),3.71–3.57(m,1H),3.55(t,J=5.2Hz,5H),3.25(t,J=0.9Hz,4H) ,3.16(t,J=5.3Hz,5H),3.08–2.93(m,4H),2.43(s,3H),2.32(s,3H),2.27(s,2H),2.11(s,2H) ,1.98–1.75(m,7H),1.75–1.55(m,4H),1.52(s,2H),1.39–1.22(m,1H),1.03(d,J=15.4Hz,6H). HR-MS(ESI):[PTX-SS-Palb+H] + Theoretical value: 1507.5473, measured value m / z = 1507.5403.

[0093] Example 8: Synthesis of PTX-SS-MK1775

[0094] PTX-SS-Np (200 mg, 0.17 mmol) was dissolved in ultra-dry CH2Cl2 (3 mL). Compound MK1775 (0.17 mmol, 82.6 mg) and ultra-dry DIEA (100 μL) were added, mixed thoroughly, and stirred at 40°C (900 rpm) for 10 hours. The reaction was monitored by TLC. After completion, PTX-SS-MK1775 (146 mg, 55.7%) was isolated by column chromatography. 1HNMR (600MHz, DMSO-d6) δ10.07(s,1H),9.23(d,J=8.5Hz,0H),8.83(s,1H),8.44(d,J=9.5Hz,0H),8.21–7.92(m,4H),7.90–7.83(m,1H),7.81–7. 68(m,2H),7.68–7.52(m,6H),7.52–7.38(m,4H),7.22(dtd,J=6.4,4.5, 4.1,2.5Hz,1H),7.00–6.90(m,2H),6.31(s,1H),5.94–5.83(m,1H),5.67 (ddt,J=17.2,10.2,6.0Hz,1H),5.59(t,J=8.6Hz,0H),5.49–5.32(m,2H ),5.26(s,1H),5.05–4.97(m,1H),4.92(dt,J=9.6,2.1Hz,1H),4.89–4.8 2(m,2H),4.68(d,J=6.0Hz,2H),4.61(d,J=12.1Hz,1H),4.40(hept,J=6 .3,5.7Hz,1H),4.33–4.21(m,3H),4.21–4.08(m,1H),4.08–3.96(m,2H).

[0095] Example 9: Synthesis of PTX-SS-MK2206

[0096] PTX-SS-Np (200 mg, 0.17 mmol) was dissolved in ultra-dry DMF (2 mL). Compound MK2206 (0.2 mmol, 88.8 mg) and ultra-dry DIEA (100 μL) were added, mixed thoroughly, and stirred at 40°C (900 rpm) for 12 hours. The reaction was monitored by TLC. After completion, PTX-SS-MK2206 (120 mg, 50%) was isolated by column chromatography. 1HNMR(600MHz,DMSO-d)δ12.62(s,1H),9.22(d,J=8.5Hz,1H),8.36(dd,J=5.7,0.7Hz,1H),8.10–8.05(m,1H ),8.00(dt,J=7.0,1.4Hz,1H),7.96(s,2H),7.94–7.83(m,3H),7.76–7.68(m,1H),7.68–7.62(m,1H),7.62– 7.51(m,2H),7.50–7.39(m,5H),7.38–7.27(m,9H),7.22(ddd,J=8.4,5.9,3.1Hz,1H),6.96(ddd,J=7.5,4. 1,0.7Hz,1H),6.31(d,J=2.4Hz,1H),5.96–5.79(m,1H),5.59(t,J=8.6Hz,1H),5.47–5.30(m,2H),4.91(dd, J=9.6,2.2Hz,1H),4.84(dd,J=11.1,6.9Hz,1H),4.61(d,J=9.5Hz,1H),4.45–4.34(m,1H),4.34–4.18(m,1 H),4.18–3.92(m,5H),3.62(dd,J=13.6,7.1Hz,1H),2.90(d,J=0.5Hz,10H),2.74(d,J=0.7Hz,5H),2.47–2. 30(m,2H),2.27(d,J=6.4Hz,3H),2.15–2.07(m,3H),2.05–1.92(m,0H),1.91–1.75(m,4H),1.65(ddd,J=13. 8,11.0,2.3Hz,1H),1.58(dd,J=15.4,9.0Hz,0H),1.55–1.47(m,3H),1.30–1.22(m,1H),1.12–0.94(m,6H). HR-MS(ESI):[PTX-SS-MK2206+H] + Theoretical value: 1467.4836, measured value m / z = 1467.4476.

[0097] Example 10: Synthesis of PTX-SS-AZD7762

[0098] PTX-SS-Np (200 mg, 0.17 mmol) was dissolved in ultra-dry CH2Cl2 (3 mL). Compound AZD7762 (0.2 mmol, 72.4 mg) and ultra-dry DIEA (100 μL) were added, mixed, and stirred at 40°C (900 rpm) for 10 hours. The reaction was monitored by TLC. After completion, PTX-SS-AZD (141 mg, 71%) was isolated by column chromatography. 1 HNMR(600MHz,DMSO-d)δ11.75(s,1H),9.06(d,J=8.6Hz,0H),6.33(s,1H),6.05–5.85(m,1H),5.58–5.30(m,2H),4.93(d d,J=9.6,2.3Hz,1H),4.75(ddt,J=11.7,7.5,4.2Hz,1H),4.65(d,J=6.6Hz,1H),4.47(s,1H),4.43–3.88(m,8H),3.81–3. 70(m,1H),3.66(dd,J=7.3,4.4Hz,1H),3.09–2.92(m,4H),2.79–2.71(m,1H),2.36(ddd,J=14.3,9.7,6.6Hz,1H),2.29( d,J=6.2Hz,2H),2.11(d,J=2.8Hz,3H),1.84(dd,J=11.5,1.5Hz,2H),1.54(s,3H),1.32–1.22(m,5H),1.11–0.98(m,5H). HR-MS(ESI):[PTX-SS-AZD+H] + Theoretical value: 1422.4303, measured value m / z = 1422.4294.

[0099] Example 11: Synthesis of PTX-SS-Cer

[0100] PTX-SS-Np (200 mg, 0.17 mmol) was dissolved in ultra-dry DMF (2 mL). Ceritinib (0.187 mmol, 112 mg) and ultra-dry DIEA (100 μL) were added, mixed thoroughly, and stirred at 40°C (900 rpm) for 10 hours. The reaction was monitored by TLC. After completion, PTX-SS-Ceritinib (200 mg, 78%) was isolated by column chromatography. 1HNMR(600MHz,DMSO-d6)δ9.46(s,1H),9.23(d,J=8.5Hz,1H),8.46(d,J=8.4Hz,1H),8.25(s,1H),8.0 2–7.91(m,3H),7.85(td,J=7.9,1.4Hz,3H),7.79–7.70(m,1H),7.69–7.60(m,2H),7.58–7.53(m,1H) ,7.52–7.41(m,5H),7.36(ddd,J=8.3,7.4,1.2Hz,1H),7.22(ddd,J=6.2,3.8,2.5Hz,1H),6.82(s,1H ),6.31(s,1H),6.13–5.78(m,1H),5.59(t,J=8.6Hz,1H),5.44(d,J=7.1Hz,1H),5.39(d,J=8.7Hz,1H) ,4.92(dd,J=9.6,2.3Hz,1H),4.66–4.49(m,2H),4.40(td,J=6.1,4.7Hz,1H),4.24(t,J=6.1Hz,2H), 4.13(dt,J=10.6,6.6Hz,3H),4.03(q,J=7.5,6.8Hz,2H),3.61(d,J=7.2Hz,1H),3.44(p,J=6.8Hz,1H) ,3.11–2.82(m,6H),2.45–2.31(m,1H),2.27(s,3H),2.21–2.07(m,6H),1.93–1.81(m,4H),1.74–1.6 3(m,3H),1.57(td,J=10.2,8.8,4.9Hz,2H),1.52(s,3H),1.27–1.14(m,12H),1.03(d,J=15.5Hz,6H). HR-MS(ESI):[PTX-SS-Cer+H] + Theoretical value: 1617.5762, measured value m / z = 1617.5260.

[0101] Example 12: Synthesis of PTX-SS-Nav

[0102] PTX-SS-Np (200 mg, 0.17 mmol) was dissolved in ultra-dry DMF (2 mL). Navitoclax (0.25 mmol, 243 mg) and ultra-dry DIEA (100 μL) were then added, mixed thoroughly, and stirred at 40°C (900 rpm) for 10 hours. The reaction was monitored by TLC. After completion, PTX-SS-Navitoclax (121 mg, 35%) was isolated by column chromatography. 1HNMR(600MHz,DMSO-d6)δ8.25(s,1H),8.18(d,J=2.2Hz,1H),8.09(dt,J=8.4,1.5Hz,2H),8.04–7.93(m,4H),7.86 (dt,J=8.6,1.6Hz,0H),7.78–7.67(m,5H),7.66–7.51(m,4H),7.51–7.39(m,5H),7.40–7.32(m,5H),7.30–7.21(m, 4H),7.22–7.16(m,1H),7.16–7.09(m,2H),7.00(dd,J=9.3,4.4Hz,1H),6.89(d,J=8.7Hz,1H),6.84(d,J=8.9Hz,2 H),6.33(s,1H),6.03–5.89(m,1H),5.70(s,2H),5.47(t,J=7.2Hz,2H),5.44–5.31(m,1H),4.93(dd,J=9.7,2.3Hz, 1H),4.65(d,J=6.8Hz,1H),4.47(s,1H),4.40(t,J=6.2Hz,0H),4.38–4.13(m,7H),4.11–4.01(m,4H),3.66(t,J=6 .6Hz,3H),3.37–3.19(m,12H),3.07–2.91(m,6H),2.85(t,J=6.4Hz,1H),2.80(s,3H),2.40–2.15(m,20H),2.11(d, J=3.7Hz,3H),2.02(d,J=2.3Hz,3H),1.99(s,1H),1.85(d,J=1.5Hz,3H),1.81–1.64(m,3H),1.54(s,3H),1.46(t,J =6.5Hz,3H),1.34–1.23(m,14H),1.20(t,J=7.1Hz,1H),1.07(d,J=3.7Hz,7H),0.99(s,7H),0.88(t,J=6.9Hz,1H). HR-MS(ESI):[PTX-SS-Nav+H] + Theoretical value: 2032.6205, measured value m / z = 2032.5682.

[0103] Example 13: Synthesis of PTX-SS-Das

[0104] PTX-SS-Np (200 mg, 0.17 mmol) was dissolved in ultra-dry DMF (2 mL). Dasatinib (0.2 mmol, 88.7 mg) and ultra-dry DIEA (100 μL) were added, mixed thoroughly, and stirred at 40°C (900 rpm) for 10 hours. The reaction was monitored by TLC. After completion, PTX-L1-Dasatinib (101 mg, 39.6%) was isolated by column chromatography. 1 HNMR(600MHz,DMSO-d6)δ11.87(s,1H),11.44(s,5H),9.83(s,5H),9.23(d,J= 8.5Hz,3H),8.44(d,J=9.6Hz,2H),8.22(s,5H),8.14–8.06(m,5H),8.04–7.95( m,11H),7.91–7.84(m,5H),7.78–7.68(m,7H),7.68–7.53(m,19H),7.52–7.36 (m,31H),7.34–7.17(m,15H),6.31(d,J=1.0Hz,5H),6.08(d,J=5.8Hz,6H),5.9 3–5.81(m,5H),5.75(s,8H),5.59(t,J=8.6Hz,2H),5.47–5.29(m,13H),4.92( dt,J=9.6,2.6Hz,6H),4.85(dd,J=9.1,6.9Hz,5H),4.61(d,J=12.4Hz,5H),4.4 8–4.37(m,5H),4.28–4.21(m,14H),4.19–4.10(m,6H),4.07–3.93(m,11H),3.6 9–3.60(m,5H),3.58–3.44(m,43H),3.03(d,J=6.2Hz,4H),2.99–2.92(m,13H). HR-MS (ESI): [PTX-SS-Das+H]+ calcd. 1548.4681, found m / z = 1548.5617.

[0105] Example 14: Synthesis of PTX-SS-Exa

[0106] PTX-SS-Np (200 mg, 0.17 mmol) was dissolved in 2 mL of ultra-dry DMF. Exatecan (0.2 mmol, 98.4 mg) and ultra-dry DIEA (100 μL) were added, mixed thoroughly, and stirred at 40°C (900 rpm) for 10 hours. The reaction was monitored by TLC. After completion, PTX-SS-Exatecan (228 mg, 90%) was isolated by column chromatography. 1HNMR(600MHz,DMSO-d6)δ9.23(d,J=8.5Hz,1H),8.42(d,J=8.5Hz,2H),8.12–8.06(m,1H),8.02–7.98(m,2H) ,7.97(d,J=7.5Hz,1H),7.90–7.83(m,2H),7.80(d,J=10.9Hz,1H),7.76–7.70(m,1H),7.65(t,J=7.7Hz,2H) ,7.62–7.52(m,2H),7.52–7.39(m,7H),7.33(s,2H),7.22(dq,J=8.7,2.5Hz,1H),6.45(s,1H),6.31(d,J=4. 2Hz,1H),5.90–5.81(m,1H),5.64–5.51(m,3H),5.47–5.35(m,5H),5.31–5.12(m,3H),4.92(dt,J=9.8,3.0Hz ,1H),4.84(s,2H),4.61(d,J=12.2Hz,1H),4.39(q,J=5.6Hz,2H),4.14(q,J=10.0,8.2Hz,4H),4.07–3.98(m ,3H),3.74–3.51(m,3H),3.18(d,J=5.5Hz,2H),3.11–2.80(m,4H),2.45–2.40(m,5H),2.33(dq,J=8.0,3.4H z,0H),2.28(d,J=8.8Hz,3H),2.25–2.17(m,1H),2.11(d,J=1.0Hz,3H),1.98–1.81(m,4H),1.81(d,J=1.4Hz ,3H),1.74–1.54(m,2H),1.52(d,J=2.4Hz,3H),1.35–1.22(m,2H),1.12–0.98(m,7H),0.88(t,J=7.3Hz,4H). HR-MS(ESI):[PTX-SS-Exa+H] + Theoretical value: 1552.4899, measured value m / z = 1552.4907.

[0107] Example 15: Synthesis of PTX-SS-Lin

[0108] PTX-SS-Np (200 mg, 0.17 mmol) was dissolved in ultra-dry DMF (2 mL). Linsitinib (0.18 mmol, 92 mg) and ultra-dry DIEA (100 μL) were added, mixed thoroughly, and stirred at 40°C (900 rpm) for 10 hours. The reaction was monitored by TLC. After completion, PTX-L1-Linsitinib (123 mg, 45.2%) was isolated by column chromatography. 1 HNMR

[0109] Example 16: Synthesis of PTX-SS-Gem

[0110] PTX-SS-Np (200 mg, 0.17 mmol) was dissolved in ultra-dry DMF (2 mL). Gemcitabine (0.2 mmol, 89.4 mg) and ultra-dry DIEA (100 μL) were added, mixed thoroughly, and stirred at 40°C (900 rpm) for 10 hours. The reaction was monitored by TLC. After completion, PTX-L1-Gemcitabine (148 mg, 47.6%) was isolated by column chromatography. 1 HNMR

[0111] Example 17: Preparation of albumin-paclitaxel nanoparticles (Nab-PTX)

[0112] 1 mg of paclitaxel was weighed and dissolved in a mixture of 64.8 μL of chloroform and 7.2 μL of ethanol as the organic phase; a 5 mg / mL aqueous solution of human serum albumin was prepared as the aqueous phase; 72 μL of the organic phase was added to 1 mL of the aqueous phase, and the suspension was sonicated continuously for 6 minutes using a 650 W probe ultrasonic disruptor (2 mm amplitude rod, 40% amplitude) with a 30-second pause and a 15-second pause sequence to obtain a nanosuspension; the suspension was then rotary evaporated on a rotary evaporator at 40°C and 533 mbar vacuum control mode for 30 minutes; and finally, the suspension was ultrafiltered using a 100 kD ultrafiltration tube at 4000 rpm for 15 minutes to obtain a nanoparticle complex.

[0113] Example 18: Preparation of Albumin-PTX-SS-T785 Nanoparticles

[0114] 1 mg of PTX-SS-T785 was weighed and dissolved in a mixture of 64.8 μL of chloroform and 7.2 μL of ethanol as the organic phase; a 5 mg / mL human serum albumin aqueous solution was prepared as the aqueous phase; 72 μL of the organic phase was added to 1 mL of the aqueous phase, and a 650 W probe ultrasonic disruptor (2 mm amplitude rod, 40% amplitude) was used to ultrasonicate for 30 seconds and pause for 15 seconds, and the nanosuspension was obtained by continuous ultrasonication for 6 minutes; then, the suspension was rotary evaporated on a rotary evaporator at 40°C and 533 mbar vacuum control mode for 30 minutes; finally, the suspension was ultrafiltered at 4000 rpm using a 100 KD ultrafiltration tube for 15 minutes to obtain a nanoparticle complex.

[0115] Example 19: Preparation of Albumin-PTX-SS-di-ABZI Nanoparticles

[0116] 1 mg of PTX-SS-di-ABZI was weighed and dissolved in a mixture of 64.8 μL of chloroform and 7.2 μL of ethanol as the organic phase; a 5 mg / mL aqueous solution of human serum albumin was prepared as the aqueous phase; 72 μL of the organic phase was added to 1 mL of the aqueous phase, and a 650 W probe ultrasonic disruptor (2 mm amplitude rod, 40% amplitude) was used to ultrasonicate for 30 seconds and pause for 15 seconds, and the nanosuspension was obtained by continuous ultrasonication for 6 minutes; then, the suspension was rotary evaporated on a rotary evaporator at 40°C and 533 mbar vacuum control mode for 30 minutes; finally, the suspension was ultrafiltered at 4000 rpm using a 100 KD ultrafiltration tube for 15 minutes to obtain a nanoparticle complex.

[0117] Example 20: Preparation of Albumin-PTX-SS-SR07 Nanoparticles

[0118] 1 mg of PTX-SS-SR07 was weighed and dissolved in a mixture of 64.8 μL of chloroform and 7.2 μL of ethanol as the organic phase; a 5 mg / mL aqueous solution of human serum albumin was prepared as the aqueous phase; 72 μL of the organic phase was added to 1 mL of the aqueous phase, and a 650 W probe ultrasonic disruptor (2 mm amplitude rod, 40% amplitude) was used to ultrasonicate for 30 seconds and pause for 15 seconds, and the nanosuspension was obtained by continuous ultrasonication for 6 minutes; then, the suspension was rotary evaporated on a rotary evaporator at 40°C and 533 mbar vacuum control mode for 30 minutes; finally, the suspension was ultrafiltered at 4000 rpm using a 100 KD ultrafiltration tube for 15 minutes to obtain a nanoparticle complex.

[0119] Example 21: Preparation of Albumin-PTX-SS-Cri Nanoparticles

[0120] 1 mg of PTX-SS-Crizotinib was weighed and dissolved in a mixture of 64.8 μL of chloroform and 7.2 μL of ethanol as the organic phase; a 5 mg / mL aqueous solution of human serum albumin was prepared as the aqueous phase; 72 μL of the organic phase was added to 1 mL of the aqueous phase, and a 650 W probe ultrasonic disruptor (2 mm amplitude rod, 40% amplitude) was used to sonicate for 30 seconds followed by a 15-second pause, and continuous sonication was performed for 6 minutes to obtain a nanosuspension; the suspension was then rotary evaporated on a rotary evaporator at 40°C and 533 mbar vacuum control mode for 30 minutes; and finally, the suspension was ultrafiltered at 4000 rpm for 15 minutes using a 100 kD ultrafiltration tube to obtain a nanoparticle complex.

[0121] Example 22: Preparation of Albumin-PTX-SS-Palb Nanoparticles

[0122] 1 mg of PTX-SS-Palbociclib was weighed and dissolved in a mixture of 64.8 μL of chloroform and 7.2 μL of ethanol as the organic phase; a 5 mg / mL aqueous solution of human serum albumin was prepared as the aqueous phase; 72 μL of the organic phase was added to 1 mL of the aqueous phase, and a 650 W probe ultrasonic disruptor (2 mm amplitude rod, 40% amplitude) was used to sonicate for 30 seconds and pause for 15 seconds, and continuous sonication was performed for 6 minutes to obtain a nanosuspension; then, the suspension was rotary evaporated on a rotary evaporator at 40°C and 533 mbar vacuum control mode for 30 minutes; finally, the suspension was ultrafiltered at 4000 rpm for 15 minutes using a 100 kD ultrafiltration tube to obtain a nanoparticle complex.

[0123] Example 23: Preparation of Albumin-PTX-SS-MK1775 Nanoparticles

[0124] 1 mg of PTX-SS-MK1775 was weighed and dissolved in a mixture of 64.8 μL of chloroform and 7.2 μL of ethanol as the organic phase; a 5 mg / mL aqueous solution of human serum albumin was prepared as the aqueous phase; 72 μL of the organic phase was added to 1 mL of the aqueous phase, and a 650 W probe-type ultrasonic disruptor (2 mm amplitude rod, 40% amplitude) was used to ultrasonicate for 30 seconds and pause for 15 seconds, and the nanosuspension was obtained by continuous ultrasonication for 6 minutes; then, the suspension was rotary evaporated on a rotary evaporator at 40°C and 533 mbar vacuum control mode for 30 minutes; finally, the suspension was ultrafiltered at 4000 rpm using a 100 KD ultrafiltration tube for 15 minutes to obtain a nanoparticle complex.

[0125] Example 24: Preparation of Albumin-PTX-SS-MK2206 Nanoparticles

[0126] 1 mg of PTX-SS-MK2206 was weighed and dissolved in a mixture of 64.8 μL of chloroform and 7.2 μL of ethanol as the organic phase; a 5 mg / mL aqueous solution of human serum albumin was prepared as the aqueous phase; 72 μL of the organic phase was added to 1 mL of the aqueous phase, and a 650 W probe-type ultrasonic disruptor (2 mm amplitude rod, 40% amplitude) was used to ultrasonicate for 30 seconds and pause for 15 seconds, and the nanosuspension was obtained by continuous ultrasonication for 6 minutes; then, the suspension was rotary evaporated on a rotary evaporator at 40°C and 533 mbar vacuum control mode for 30 minutes; finally, the suspension was ultrafiltered at 4000 rpm using a 100 KD ultrafiltration tube for 15 minutes to obtain a nanoparticle complex.

[0127] Example 25: Preparation of Albumin-PTX-SS-AZD Nanoparticles

[0128] 1 mg of PTX-SS-AZD7762 was weighed and dissolved in a mixture of 64.8 μL of chloroform and 7.2 μL of ethanol as the organic phase; a 5 mg / mL aqueous solution of human serum albumin was prepared as the aqueous phase; 72 μL of the organic phase was added to 1 mL of the aqueous phase, and a 650 W probe ultrasonic disruptor (2 mm amplitude rod, 40% amplitude) was used to ultrasonicate for 30 seconds and pause for 15 seconds, and the nanosuspension was obtained by continuous ultrasonication for 6 minutes; then, the suspension was rotary evaporated on a rotary evaporator at 40°C and 533 mbar vacuum control mode for 30 minutes; finally, the suspension was ultrafiltered at 4000 rpm using a 100 KD ultrafiltration tube for 15 minutes to obtain a nanoparticle complex.

[0129] Example 26: Preparation of Albumin-Cer Nanoparticles

[0130] 1 mg of PTX-SS-Ceritinib was weighed and dissolved in a mixture of 64.8 μL of chloroform and 7.2 μL of ethanol as the organic phase; a 5 mg / mL aqueous solution of human serum albumin was prepared as the aqueous phase; 72 μL of the organic phase was added to 1 mL of the aqueous phase, and a 650 W probe ultrasonic disruptor (2 mm amplitude rod, 40% amplitude) was used to ultrasonicate for 30 seconds and pause for 15 seconds, and the nanosuspension was obtained by continuous ultrasonication for 6 minutes; then, the suspension was rotary evaporated on a rotary evaporator at 40°C and 533 mbar vacuum control mode for 30 minutes; finally, the suspension was ultrafiltered at 4000 rpm using a 100 KD ultrafiltration tube for 15 minutes to obtain a nanoparticle complex.

[0131] Example 27: Preparation of Albumin-PTX-SS-Nav Nanoparticles

[0132] 1 mg of PTX-SS-Navitoclax was weighed and dissolved in a mixture of 64.8 μL of chloroform and 7.2 μL of ethanol as the organic phase; a 5 mg / mL aqueous solution of human serum albumin was prepared as the aqueous phase; 72 μL of the organic phase was added to 1 mL of the aqueous phase, and a 650 W probe ultrasonic disruptor (2 mm amplitude rod, 40% amplitude) was used to ultrasonicate for 30 seconds and pause for 15 seconds, and the suspension was continuously ultrasonicated for 6 minutes to obtain a nanosuspension; the suspension was then rotary evaporated on a rotary evaporator at 40°C and 533 mbar vacuum control mode for 30 minutes; and finally, the suspension was ultrafiltered at 4000 rpm using a 100 KD ultrafiltration tube for 15 minutes to obtain a nanoparticle complex.

[0133] Example 28: Preparation of Albumin-PTX-SS-Das Nanoparticles

[0134] 1 mg of PTX-SS-Dasatinib was weighed and dissolved in a mixture of 64.8 μL of chloroform and 7.2 μL of ethanol as the organic phase; a 5 mg / mL aqueous solution of human serum albumin was prepared as the aqueous phase; 72 μL of the organic phase was added to 1 mL of the aqueous phase, and a 650 W probe ultrasonic disruptor (2 mm amplitude rod, 40% amplitude) was used to ultrasonicate for 30 seconds and pause for 15 seconds, and the suspension was obtained by continuous ultrasonication for 6 minutes; the suspension was then rotary evaporated on a rotary evaporator at 40°C and 533 mbar vacuum control mode for 30 minutes; and finally, the suspension was ultrafiltered at 4000 rpm for 15 minutes using a 100 KD ultrafiltration tube to obtain a nanoparticle complex.

[0135] Example 29: Preparation of Albumin-PTX-SS-Exa Nanoparticles

[0136] 1 mg of PTX-SS-Exatecan was weighed and dissolved in a mixture of 64.8 μL of chloroform and 7.2 μL of ethanol as the organic phase; a 5 mg / mL aqueous solution of human serum albumin was prepared as the aqueous phase; 72 μL of the organic phase was added to 1 mL of the aqueous phase; a 650 W probe ultrasonic disruptor (2 mm amplitude rod, 40% amplitude) was used to ultrasonicate for 30 seconds and pause for 15 seconds, and continuous ultrasonication was performed for 6 minutes to obtain a nanosuspension; then, the suspension was rotary evaporated on a rotary evaporator at 40°C and 533 mbar vacuum control mode for 30 minutes; finally, the suspension was ultrafiltered at 4000 rpm using a 100 KD ultrafiltration tube for 15 minutes to obtain a nanoparticle complex.

[0137] Example 30: Preparation of Albumin-PTX-SS-Lin Nanoparticles

[0138] 1 mg of PTX-SS-Linsitinib was weighed and dissolved in a mixture of 64.8 μL of chloroform and 7.2 μL of ethanol as the organic phase; a 5 mg / mL aqueous solution of human serum albumin was prepared as the aqueous phase; 72 μL of the organic phase was added to 1 mL of the aqueous phase; a 650 W probe ultrasonic disruptor (2 mm amplitude rod, 40% amplitude) was used to ultrasonicate for 30 seconds and pause for 15 seconds, and continuous ultrasonication was performed for 6 minutes to obtain a nanosuspension; then, the suspension was rotary evaporated on a rotary evaporator at 40°C and 533 mbar vacuum control mode for 30 minutes; finally, the suspension was ultrafiltered at 4000 rpm using a 100 KD ultrafiltration tube for 15 minutes to obtain a nanoparticle complex.

[0139] Example 31: Preparation of Albumin-PTX-SS-Gem Nanoparticles

[0140] 1 mg of PTX-SS-Gemcitabine was weighed and dissolved in a mixture of 64.8 μL of chloroform and 7.2 μL of ethanol as the organic phase; a 5 mg / mL aqueous solution of human serum albumin was prepared as the aqueous phase; 72 μL of the organic phase was added to 1 mL of the aqueous phase; a 650 W probe ultrasonic disruptor (2 mm amplitude rod, 40% amplitude) was used to ultrasonicate for 30 seconds and pause for 15 seconds, and continuous ultrasonication was performed for 6 minutes to obtain a nanosuspension; then, the suspension was rotary evaporated on a rotary evaporator at 40°C and 533 mbar vacuum control mode for 30 minutes; finally, the suspension was ultrafiltered at 4000 rpm using a 100 KD ultrafiltration tube for 15 minutes to obtain a nanoparticle complex.

[0141] Example 32: Preparation of Albumin-PTX-SS-Cri@MK1775 Nanoparticles

[0142] 0.5 mg of PTX-SS-Cri and PTX-SS-MK1775 were weighed respectively and dissolved in a mixture of 32.4 μL of chloroform and 3.6 μL of ethanol as organic phases A and B; a 5 mg / mL human serum albumin aqueous solution was prepared as the aqueous phase; the two organic phases A and B were added to 1 mL of the aqueous phase in a ratio of 1:1; a 650W probe-type ultrasonic disruptor (2 mm amplitude rod, 40% amplitude) was used to ultrasonicate for 30 seconds and pause for 15 seconds, and continuous ultrasonication was performed for 6 minutes to obtain a nanosuspension; then, the mixture was rotary evaporated on a rotary evaporator for 30 minutes, using a vacuum control mode of 40°C and 533 mbar; finally, the mixture was ultrafiltered at 4000 rpm using a 100KD ultrafiltration tube for 15 minutes to obtain a dual-drug nanoparticle complex.

[0143] Example 33: Preparation of Albumin-PTX-SS-Exa@MK1775 Nanoparticles

[0144] 0.5 mg of PTX-SS-Exa and PTX-SS-MK1775 were weighed respectively and dissolved in a mixture of 32.4 μL of chloroform and 3.6 μL of ethanol as organic phases A and B; a 5 mg / mL human serum albumin aqueous solution was prepared as the aqueous phase; the two organic phases A and B were added to 1 mL of the aqueous phase in a ratio of 1:1; a 650W probe-type ultrasonic disruptor (2 mm amplitude rod, 40% amplitude) was used to ultrasonicate for 30 seconds and pause for 15 seconds, and continuous ultrasonication was performed for 6 minutes to obtain a nanosuspension; then, the mixture was rotary evaporated on a rotary evaporator for 30 minutes, using a vacuum control mode of 40°C and 533 mbar; finally, the mixture was ultrafiltered at 4000 rpm using a 100KD ultrafiltration tube for 15 minutes to obtain a dual-drug nanoparticle complex.

[0145] Example 34: Preparation of Albumin-PTX-SS-Pal@MK2206 Nanoparticles

[0146] 0.5 mg of PTX-SS-Palbociclib and PTX-SS-MK2206 were weighed separately and dissolved in a mixture of 32.4 μL of chloroform and 3.6 μL of ethanol as organic phases A and B; a 5 mg / mL human serum albumin aqueous solution was prepared as the aqueous phase; the two organic phases A and B were added to 1 mL of the aqueous phase in a ratio of 1:1; a 650 W probe-type ultrasonic disruptor (2 mm amplitude rod, 40% amplitude) was used to ultrasonicate for 30 seconds and pause for 15 seconds, and continuous ultrasonication was performed for 6 minutes to obtain a nanosuspension; then, the suspension was rotary evaporated on a rotary evaporator for 30 minutes using a vacuum control mode of 40°C and 533 mbar; finally, the suspension was ultrafiltered at 4000 rpm using a 100 KD ultrafiltration tube for 15 minutes to obtain a dual-drug nanoparticle complex.

[0147] Example 35: Preparation of Albumin-PTX-SS-MK2206@MK1775 Nanoparticles

[0148] 0.5 mg of PTX-SS-MK2206 and PTX-SS-MK1775 were weighed respectively and dissolved in a mixture of 32.4 μL of chloroform and 3.6 μL of ethanol as organic phases A and B; a 5 mg / mL human serum albumin aqueous solution was prepared as the aqueous phase; the two organic phases A and B were added to 1 mL of the aqueous phase in a ratio of 1:1; a 650W probe-type ultrasonic disruptor (2 mm amplitude rod, 40% amplitude) was used to ultrasonicate for 30 seconds and pause for 15 seconds, and continuous ultrasonication was performed for 6 minutes to obtain a nanosuspension; then, the mixture was rotary evaporated on a rotary evaporator for 30 minutes using a vacuum control mode of 40°C and 533 mbar; finally, the mixture was ultrafiltered at 4000 rpm using a 100KD ultrafiltration tube for 15 minutes to obtain a dual-drug nanoparticle complex.

[0149] Example 36: Preparation of Albumin-PTX-SS-Das@MK1775 Nanoparticles

[0150] 0.5 mg of PTX-SS-Dasatinib and PTX-SS-MK1775 were weighed separately and dissolved in a mixture of 32.4 μL of chloroform and 3.6 μL of ethanol as organic phases A and B; a 5 mg / mL human serum albumin aqueous solution was prepared as the aqueous phase; the two organic phases A and B were added to 1 mL of the aqueous phase in a ratio of 1:1; a 650W probe-type ultrasonic disruptor (2 mm amplitude rod, 40% amplitude) was used to ultrasonicate for 30 seconds and pause for 15 seconds, and continuous ultrasonication was performed for 6 minutes to obtain a nanosuspension; then, the mixture was rotary evaporated on a rotary evaporator for 30 minutes using a vacuum control mode of 40°C and 533 mbar; finally, the mixture was ultrafiltered at 4000 rpm using a 100KD ultrafiltration tube for 15 minutes to obtain a dual-drug nanoparticle complex.

[0151] Example 37: Preparation of Albumin-PTX-SS-Exa@Cri Nanoparticles

[0152] 0.5 mg of PTX-SS-Exatecan and PTX-SS-Crizotinib were weighed respectively and dissolved in a mixture of 32.4 μL of chloroform and 3.6 μL of ethanol as organic phases A and B; a 5 mg / mL human serum albumin aqueous solution was prepared as the aqueous phase; the two organic phases A and B were added to 1 mL of the aqueous phase in a ratio of 1:1; a 650W probe-type ultrasonic disruptor (2 mm amplitude rod, 40% amplitude) was used to ultrasonicate for 30 seconds and pause for 15 seconds, and continuous ultrasonication was performed for 6 minutes to obtain a nanosuspension; then, the mixture was rotary evaporated on a rotary evaporator for 30 minutes using a vacuum control mode of 40°C and 533 mbar; finally, the mixture was ultrafiltered at 4000 rpm using a 100KD ultrafiltration tube for 15 minutes to obtain a dual-drug nanoparticle complex.

[0153] Example 38: Preparation of Albumin-PTX-SS-Exa@Das Nanoparticles

[0154] 0.5 mg of PTX-SS-Exatecan and PTX-SS-Dasatinib were weighed separately and dissolved in a mixture of 32.4 μL of chloroform and 3.6 μL of ethanol as organic phases A and B; a 5 mg / mL human serum albumin aqueous solution was prepared as the aqueous phase; the two organic phases A and B were added to 1 mL of the aqueous phase in a ratio of 1:1; a 650W probe-type ultrasonic disruptor (2 mm amplitude rod, 40% amplitude) was used to ultrasonicate for 30 seconds and pause for 15 seconds, and the nanosuspension was obtained by continuous ultrasonication for 6 minutes; then, the suspension was rotary evaporated on a rotary evaporator for 30 minutes using a vacuum control mode of 40°C and 533 mbar; finally, the mixture was ultrafiltered at 4000 rpm using a 100KD ultrafiltration tube for 15 minutes to obtain a dual-drug nanoparticle complex.

[0155] Example 39: Preparation of Albumin-PTX-SS-MK2206@MK1775@T785 Nanoparticles

[0156] 0.33 mg of PTX-SS-MK2206, PTX-SS-MK1775 and PTX-L-T785 were weighed respectively and dissolved in a mixture of 21.6 μL of chloroform and 2.4 μL of ethanol as three organic phases A, B and C; a 5 mg / mL human serum albumin aqueous solution was prepared as the aqueous phase; the three organic phases A, B and C were added to 1 mL of the aqueous phase in a ratio of 1:1:1; a 650 W probe-type ultrasonic disruptor (2 mm amplitude rod, 40% amplitude) was used to ultrasonicate for 30 seconds and pause for 15 seconds, and continuous ultrasonication was performed for 6 minutes to obtain a nanosuspension; then, the mixture was rotary evaporated on a rotary evaporator for 30 minutes, using a vacuum control mode of 40°C and 533 mbar; finally, the mixture was ultrafiltered at 4000 rpm using a 100KD ultrafiltration tube for 15 minutes to obtain a three-drug nanoparticle complex.

[0157] Example 40: Preparation of Albumin-PTX-SS-Cri@Cer@SR07 Nanoparticles

[0158] 0.33 mg of PTX-SS-Crizotinib, PTX-SS-Ceritinib and PTX-SS-SR07 were weighed respectively and dissolved in a mixture of 21.6 μL chloroform and 2.4 μL ethanol as three organic phases A, B and C; a 5 mg / mL human serum albumin aqueous solution was prepared as the aqueous phase; the three organic phases A, B and C were added to 1 mL of the aqueous phase in a ratio of 1:1:1; a 650W probe-type ultrasonic disruptor (2 mm amplitude rod, 40% amplitude) was used to ultrasonicate for 30 seconds and pause for 15 seconds, and continuous ultrasonication was performed for 6 minutes to obtain a nanosuspension; then, the mixture was rotary evaporated on a rotary evaporator for 30 minutes, using a vacuum control mode of 40°C and 533 mbar; finally, the three-drug nanoparticle complex was obtained by ultrafiltration at 4000 rpm using a 100KD ultrafiltration tube for 15 minutes.

[0159] Example 41: Preparation of Albumin-PTX-SS-MK2206@Pal@T785 Nanoparticles

[0160] 0.33 mg of PTX-SS-MK2206, PTX-SS-Palbociclib and PTX-SS-T785 were weighed respectively and dissolved in a mixture of 21.6 μL of chloroform and 2.4 μL of ethanol as three organic phases A, B and C; a 5 mg / mL human serum albumin aqueous solution was prepared as the aqueous phase; the three organic phases A, B and C were added to 1 mL of the aqueous phase in a ratio of 1:1:1; a 650 W probe-type ultrasonic disruptor (2 mm amplitude rod, 40% amplitude) was used to ultrasonicate for 30 seconds and pause for 15 seconds, and continuous ultrasonication was performed for 6 minutes to obtain a nanosuspension; then, the mixture was rotary evaporated on a rotary evaporator for 30 minutes using a vacuum control mode of 40°C and 533 mbar; finally, the mixture was ultrafiltered at 4000 rpm using a 100KD ultrafiltration tube for 15 minutes to obtain a three-drug nanoparticle complex.

[0161] Example 42: Preparation of Albumin-PTX-SS-Pal@Cer@SR07 Nanoparticles

[0162] 0.33 mg of PTX-SS-Palbociclib, PTX-SS-Ceritinib and PTX-SS-SR07 were weighed respectively and dissolved in a mixture of 21.6 μL chloroform and 2.4 μL ethanol as three organic phases A, B and C; a 5 mg / mL human serum albumin aqueous solution was prepared as the aqueous phase; the three organic phases A, B and C were added to 1 mL of the aqueous phase in a ratio of 1:1:1; a 650W probe-type ultrasonic disruptor (2 mm amplitude rod, 40% amplitude) was used to ultrasonicate for 30 seconds and pause for 15 seconds, and continuous ultrasonication was performed for 6 minutes to obtain a nanosuspension; then, the mixture was rotary evaporated on a rotary evaporator for 30 minutes, using a vacuum control mode of 40°C and 533 mbar; finally, the mixture was ultrafiltered at 4000 rpm using a 100KD ultrafiltration tube for 15 minutes to obtain a three-drug nanoparticle complex.

[0163] Example 43: Preparation of Albumin-PTX-SS-Pal@Cer@T785 Nanoparticles

[0164] 0.33 mg of PTX-SS-Palbociclib, PTX-SS-Ceritinib and PTX-SS-T785 were weighed respectively and dissolved in a mixture of 21.6 μL chloroform and 2.4 μL ethanol as three organic phases A, B and C; a 5 mg / mL human serum albumin aqueous solution was prepared as the aqueous phase; the three organic phases A, B and C were added to 1 mL of the aqueous phase in a ratio of 1:1:1; a 650W probe-type ultrasonic disruptor (2 mm amplitude rod, 40% amplitude) was used to ultrasonicate for 30 seconds and pause for 15 seconds, and continuous ultrasonication was performed for 6 minutes to obtain a nanosuspension; then, the mixture was rotary evaporated on a rotary evaporator for 30 minutes using a vacuum control mode of 40°C and 533 mbar; finally, the mixture was ultrafiltered at 4000 rpm using a 100KD ultrafiltration tube for 15 minutes to obtain a three-drug nanoparticle complex.

[0165] Example 44: Preparation of Albumin-PTX-SS-Cer@Cri@T785 Nanoparticles

[0166] 0.33 mg of PTX-SS-Ceritinib, PTX-SS-Crizotinib and PTX-SS-T785 were weighed respectively and dissolved in a mixture of 21.6 μL chloroform and 2.4 μL ethanol as three organic phases A, B and C; a 5 mg / mL human serum albumin aqueous solution was prepared as the aqueous phase; the three organic phases A, B and C were added to 1 mL of the aqueous phase in a ratio of 1:1:1; a 650W probe-type ultrasonic disruptor (2 mm amplitude rod, 40% amplitude) was used to ultrasonicate for 30 seconds and pause for 15 seconds, and continuous ultrasonication was performed for 6 minutes to obtain a nanosuspension; then, the mixture was rotary evaporated on a rotary evaporator for 30 minutes, using a vacuum control mode of 40°C and 533 mbar; finally, ultrafiltration was performed at 4000 rpm using a 100KD ultrafiltration tube for 15 minutes to obtain a three-drug nanoparticle complex.

[0167] Example 45: Preparation of Albumin-Exa@MK2206@SR07 Nanoparticles

[0168] 0.33 mg of PTX-SS-Exatecan, PTX-SS-MK2206 and PTX-SS-SR07 were weighed respectively and dissolved in a mixture of 21.6 μL chloroform and 2.4 μL ethanol as organic phases A, B and C; a 5 mg / mL human serum albumin aqueous solution was prepared as the aqueous phase; the three organic phases A, B and C were added to 1 mL of the aqueous phase in a ratio of 1:1:1; a 650W probe-type ultrasonic disruptor (2 mm amplitude rod, 40% amplitude) was used to ultrasonicate for 30 seconds and pause for 15 seconds, and the nanosuspension was obtained by continuous ultrasonication for 6 minutes; then the mixture was rotary evaporated on a rotary evaporator for 30 minutes, using a vacuum control mode of 40°C and 533 mbar; finally, the three-drug nanoparticle complex was obtained by ultrafiltration at 4000 rpm using a 100KD ultrafiltration tube for 15 minutes.

[0169] Experimental Example 1: Hydration Particle Size and Surface Potential of Albumin-Paclitaxel-Derived Twin-Drug Nanoparticles

[0170] The nanoparticles obtained in Examples 17-45 were diluted 10-fold with deionized water, and the particle size, distribution, and PDI of the albumin-paclitaxel and albumin-paclitaxel twin-drug nanoparticles were measured using a Malvern Zetasizer Nano ZS nanoparticle size potentiometer. The surface potential was also measured. The specific results are shown in Table 1:

[0171] Table 1 Test results of albumin-paclitaxel-derived twin-drug nanoparticles

[0172] The particle size and surface potential of albumin-paclitaxel twin-drug nanoparticles are shown in Figure 1 and Table 1. The results show that, except for the hydrated particle size of Nab-PTX at 162.1 nm, the particle size of the other albumin-bound paclitaxel-derived twin-drugs are all between 90 and 140 nm, with negative surface potentials and PDIs less than 0.24. However, the hydrated particle sizes of nanoparticles combining different paclitaxel twin-drug compounds vary greatly. For example, the hydrated particle sizes of nanoparticles Nab-PTX-SS-Linsitinib, Nab-PTX-SS-Dasatinib, and Nab-PTX-SS-MK2206@MK1775@T785 are less than 100 nm, while the hydrated particle sizes of Nab-PTX-SS-Pal@Cer@SR07 and Nab-PTX-SS-Pal@Cri@T785 are close to 140 nm. The smaller particle size means that the nanoparticles can more easily pass through the blood vessel wall and accumulate in tumor tissue, thereby improving the distribution and targeting ability of the nanoparticles in the body. Nanoparticles with larger particle sizes may be quickly cleared by the body system and fail to exert their therapeutic effects well.

[0173] Experimental Example 2: Drug loading rate of albumin-paclitaxel-derived twin-drug nanoparticles

[0174] Preferably, this article investigated the encapsulation efficiency and drug loading of PTX-SS-T785 in albumin-PTX-SS-T785 nanoparticles. 1.2 mg, 1.5 mg, and 2 mg of PTX-SS-T785 were weighed and dissolved in 1 mL of DMSO, respectively. High-performance liquid chromatography (HPLC) was used to generate a standard curve. 1.4 mg of PTX-SS-T785 was weighed and albumin nanoparticles were prepared using the protocol in Example 16. 12.5 μL of the sample was added to DMSO, sonicated for 10 minutes, and centrifuged at 8000 rpm for 5 minutes. The supernatant was collected and the drug mass in the nanoparticles was measured. 50 μL and 100 μL of the nanoparticles prepared in Example 16 were then added to DMSO to a volume of 500 μL. After sonication for 10 minutes, the sample was centrifuged at 8000 rpm for 5 minutes. The supernatant was collected and injected into HPLC for analysis, using a 10 μL injection volume. Chromatographic conditions: A Waters XBridge-C18 column (4.6 × 250 mm, 5 μm) was used, with a mobile phase of acetonitrile:water (65 / 35, v / v), a detection wavelength of 254 nm, and a column temperature of 40°C. Drug encapsulation efficiency was calculated as follows: Encapsulation efficiency (%) = drug mass in nanoparticles / actual dose × 100%; Drug loading (%) = drug mass in nanoparticles / total mass of nanoparticles × 100%;

[0175] The specific results are as follows: the total mass of albumin-PTX-SS-T785 nanoparticles is 19.433, the mass of nanoparticles PTX-L-T785 is 1 mg, and it is calculated that the encapsulation efficiency is 71.43% and the drug loading is 5.15%.

[0176] Test Example 3: Cytotoxicity Assay

[0177] The CCK8 method was used to investigate the cytotoxicity of PTX combined with anti-tumor drugs (chemotherapeutic drugs, small molecule inhibitors, etc.) on pancreatic cancer cells such as BxPC-3, CFPAC-1, and PATU-8988T: cells (1000 cells / well) were seeded into 96-well plates and cultured overnight. Then, different concentrations of paclitaxel, different concentrations of anti-tumor drugs, and paclitaxel (1 μM)-different concentrations of anti-tumor drugs were added. After 72 hours, 10 μL of CCK8 working solution was added to each well. The cells were incubated in a cell culture incubator for 1-2 hours. The absorbance of each well was detected (450 nm) using an enzyme reader and the viability was calculated. Finally, the combination index (CI) of paclitaxel and various anti-tumor drugs was calculated using CompuSyn software.

[0178] The drug molecules are Linsitinib, Gemcitabine, Crizotinb, Palbociclib, MK1775, MK2206, AZD7762, Ceritinib, Navitoclax, Dasatinib and Exatecan

[0179] Figure 2a shows the cytotoxicity of paclitaxel combined with anti-tumor drugs. It can be seen that individual anti-tumor drugs have inconsistent inhibitory effects on tumor cell proliferation. Crizotinb, Linsitinib, Dasatinib, MK2206, and Exatecan showed weak inhibitory effects on both BxPC-3 and PATU-8988T cell lines, while MK1775 and Gemcitabine showed stronger inhibitory effects. AZD7762 and Navitoclax also showed strong inhibitory effects on BxPC-3 cell proliferation. The combination of paclitaxel and anti-tumor drugs significantly altered the inhibitory effect of the paclitaxel twin-drug compound on tumor cell proliferation. As can be seen from Figure 2a, the combination of paclitaxel with four drug molecules, namely Crizotinb, Linsitinib, Dasatinib, MK2206 and Exatecan, can significantly improve the proliferation inhibitory effect of paclitaxel twin-drug compounds on BxPC-3 and PATU-8988T cells; when used in combination with Navitoclax, the drug molecules significantly enhanced the proliferation inhibitory effect on PATU-8988T cells; while the paclitaxel twin-drug compounds prepared with MK1775 or Gemcitabine had no significant improvement in the proliferation inhibitory effect on tumor cells; at the same time, it can also be seen that the combination of paclitaxel and anti-tumor drugs has a stronger inhibitory effect on the proliferation of PATU-8988T cells than on the proliferation of BxPC-3 cells.

[0180] As shown in Figure 2b, it can be found that the synergistic indexes of different drugs with paclitaxel are different. Among them, the synergistic indexes of paclitaxel when used in combination with Linsitinib, Navitoclax, MK2206, and AZD7762 drug molecules are all significantly greater than 1, and are basically consistent in the two cell types. When used in combination with Dasatinib, the synergistic index of paclitaxel on the inhibition of PATU-8988T cell proliferation is also significantly greater than 1, indicating that the effect of paclitaxel combined with these drug molecules is significantly better than that of a single drug; while the synergistic indexes of Palbociclib and Ceritinib with paclitaxel are relatively low, and the effect of the two drugs acting together is equal to or even worse than the sum of the effects of the two drugs acting alone.

[0181] The above results show that the combination of paclitaxel and a single anti-tumor drug can significantly improve the inhibitory effect on cell proliferation, among which the combination of paclitaxel with Linsitinib, Navitoclax, MK2206, and Dasatinib has a better effect.

[0182] Experimental Example 4: In vivo anti-tumor effect of albumin-paclitaxel twin drugs

[0183] KPC pancreatic cancer cell suspension was mixed with matrigel at a ratio of 1:1 and injected into the right axilla of C57BL / 6 mice until the tumor volume reached 100 mm. 3 The tumor-bearing mice were randomly divided into groups of 6 each. The treatments shown in Table 2 were performed respectively. The tumor volume and mouse body weight were recorded every two days. The tumor volume change graph and survival curve were plotted. The tumor volume change data are shown in Table 3, and the weight change graph of the mice after treatment with PBS, Nab-PTX, Nab-PTX-SS-T785, Nab-PTX-SS-Navitoclax, Nab-PTX-SS-Exatecan, and Nab-PTX-SS-Das@MK1775 is shown in Figure 4.

[0184] Table 2 Mouse experiment table

[0185] Table 3 Changes in tumor volume in mice

[0186] The changes in tumor volume after intravenous administration in pancreatic cancer-bearing C57 mice are shown in Table 3 and Figure 3. Compared with the control group (PBS), the antitumor effect of albumin-paclitaxel nanoparticles prepared with paclitaxel alone was not obvious. The twin-drug nanoparticles prepared with paclitaxel and multiple antitumor drugs had a significant inhibitory effect on tumor growth. However, there were certain differences in the antitumor effects of the albumin-paclitaxel twin-drug nanoparticles, including single-drug, double-drug or triple-drug nanoparticles. Among them, the single-drug nanoparticles Nab-PTX-SS-Linsitinib, Nab-PT X-SS-T785, Nab-PTX-SS-Navitoclax, Nab-PTX-SS-Dasatinib, Nab-PTX-SS-MK2206, Nab-PTX-SS-Exatecan and the nanoparticles containing the three drugs, Nab-PTX-SS-Cri@Cer@SR07 and Nab-PTX-SS-Exa@MK2206@SR07, had a significant inhibitory effect on tumor growth, and the tumor volume was less than 200 mm on the 22nd day. 3Among them, the nanoparticle Nab-PTX-SS-Linsitinib had the best effect in inhibiting tumor growth. Combined with the hydrated particle sizes of different nanoparticles in Experimental Example 1, it can be found that the hydrated particle sizes of nanoparticles Nab-PTX-SS-Linsitinib, Nab-PTX-SS-T785, Nab-PTX-SS-Navitoclax, Nab-PTX-SS-Dasatinib, Nab-PTX-SS-MK2206, and Nab-PTX-SS-Exatecan are 90.5 nm, 118.5 nm, 110.7 nm, 93.26 nm, 114.9 nm, and 105.2 nm, respectively, which are between 90 and 120 nm. At the same time, combined with the results of Experimental Example 2, it can be concluded that the smaller the hydrated particle size of the nanoparticles, the better the anti-tumor growth effect of the nanoparticles when there is a synergistic effect between the paclitaxel and the drug molecules in the paclitaxel twin-drug compound. Among them, the anti-tumor effect of the nanoparticles Nab-PTX-SS-Linsitinib is the best.

[0187] As can be seen from Figure 4, there is no significant change in the body weight of tumor-bearing mice after treatment with different nanoparticles. The reason may be that the weight of the tumor accounts for a small proportion of the body weight of the tumor-bearing mice, and changes in the size of the tumor will not cause significant changes in the body weight of the tumor-bearing mice.

[0188] The above embodiments are intended to illustrate the present invention only and are not intended to limit the present invention. Persons skilled in the art may make various changes and modifications without departing from the spirit and scope of the present invention. Therefore, all equivalent technical solutions also fall within the scope of the present invention, and the scope of patent protection of the present invention shall be defined by the claims.

[0189] The embodiments and / or implementation methods described above are only used to illustrate the preferred embodiments and / or implementation methods for realizing the technology of the present invention, and do not impose any form of limitation on the implementation methods of the technology of the present invention. Any person skilled in the art may make slight changes or modifications to other equivalent embodiments without departing from the scope of the technical means disclosed in the content of the present invention, but they should still be regarded as technologies or embodiments that are essentially the same as the present invention.

[0190] This article uses specific examples to illustrate the principles and implementation methods of this application. The description of the above embodiments is only used to help understand the method and core ideas of this application. The above is only the preferred implementation method of this application. It should be pointed out that due to the limitations of textual expression, there are objectively infinite specific structures. For ordinary technicians in this technical field, without departing from the principles of this application, they can also make several improvements, modifications or changes, and can also combine the above technical features in an appropriate manner; these improvements, modifications, changes or combinations, or the direct application of the inventive concept and technical solution to other occasions without improvement, should be regarded as the scope of protection of this application.

Claims

1. A paclitaxel twin drug compound, characterized in that: It has the following structural formula: The R is a drug molecule, including any one of a cytotoxic drug, a small molecule targeted drug, an immune checkpoint inhibitor, a hormone drug, and an immune agonist; the structure of the Linker is any one of the following: wherein n is any integer from 1 to 14, m is any integer from 1 to 14, R1 is at least one of a hydrazone bond, a hydrazide bond, a disulfide bond, a thioether bond, a diselenide bond, a selenoether bond, a thioketal bond and a Michael adduct, R2 is at least one of a hydrazone bond, a hydrazide bond, a disulfide bond, a thioether bond, a diselenide bond, a selenoether bond, a thioketal bond and a Michael adduct, R3 is at least one of a hydrazone bond, a hydrazide bond, a disulfide bond, a thioether bond, a diselenide bond, a selenoether bond, a thioketal bond and a Michael adduct, and R4 is at least one of a hydrazone bond, a hydrazide bond, a disulfide bond, a thioether bond, a diselenide bond, a selenoether bond, a thioketal bond and a Michael adduct.

2. A paclitaxel twin drug compound according to claim 1, characterized in that: The drug molecule is any one of T785, di-ABZI, SR07, Crizotinb, Palbociclib, MK1775, MK2206, AZD7762, Ceritinib, Navitoclax, Linsitinib, Gemcitabine, Dasatinib and Exatecan; the structure of the Linker is as follows:

3. A paclitaxel twin drug compound according to claim 2, characterized in that: The drug molecule is any one of Linsitinib, MK2206, Navitoclax, and Dasatinib.

4. A method for preparing the paclitaxel twin-drug compound according to claim 1 or 2, characterized in that: include: The drug molecule, linker and paclitaxel are reacted to obtain a paclitaxel twin drug compound.

5. The method for preparing the paclitaxel twin-drug compound according to claim 4, characterized in that: The linker molecule reacts with a solvent to prepare an intermediate, and then the intermediate reacts with a drug molecule in an organic solvent to prepare a paclitaxel twin drug compound.

6. The method for preparing a paclitaxel twin-drug compound according to claim 5, characterized in that: A solvent is used in the chemical reaction, and the solvent is one or more of dichloromethane, chloroform, 1,2-dichloroethane, tetrahydrofuran, acetonitrile, N,N-dimethylformamide, 1,4-dioxane, dimethyl sulfoxide, benzene, toluene, xylene, chlorobenzene, and o-dichlorobenzene; or, an alkaline reagent is used in the chemical reaction, and the alkaline reagent is one or more of alkali metal hydroxide, alkaline earth metal hydroxide, alkali metal or alkaline earth metal carbonate, alkali metal or alkaline earth metal bicarbonate, triethylamine, tributylamine, trioctylamine, N,N-diisopropylethylamine, pyridine, 4-dimethylaminopyridine, piperidine, N-methylmorpholine, N-methylpiperidine, tetrahydropyrrole, triethylenediamine, and tetrabutylammonium hydroxide; or, a condensing agent is used in the chemical reaction, and the condensing agent is one or more of EDCI, DCC, and DIC.

7. A paclitaxel twin-drug nanoparticle, characterized in that: It comprises a paclitaxel twin-drug compound as described in any one of claims 1 to 3.

8. The paclitaxel twin-drug nanoparticle according to claim 7, characterized in that: The paclitaxel twin-drug nanoparticles also include albumin, and the paclitaxel twin-drug compound is bonded to the albumin.

9. The paclitaxel twin-drug nanoparticle according to claim 8, characterized in that: The albumin includes at least one of bovine serum albumin, human serum albumin, ovalbumin and recombinant human serum albumin; or, the bonding is a non-covalent bond; or, the paclitaxel twin-drug nanoparticles include at least one paclitaxel twin-drug compound.

10. The paclitaxel twin-drug nanoparticle according to claim 9, characterized in that: The albumin includes human serum albumin.

11. The paclitaxel twin-drug nanoparticle according to claim 9, characterized in that: The paclitaxel twin-drug nanoparticles include 1, 2, 3, 4 or 5 paclitaxel twin-drug compounds.

12. The paclitaxel twin-drug nanoparticle according to claim 11, characterized in that: The paclitaxel twin-drug compound includes any one of Linsitinib, MK2206, Navitoclax, and Dasatinib drug molecules.

13. The paclitaxel twin-drug nanoparticle according to claim 12, characterized in that: The hydrated particle size of the paclitaxel twin-drug nanoparticles is 90-140 nm, the surface potential is negative, and the PDI is less than 0.

24.

14. A method for preparing paclitaxel twin-drug nanoparticles, characterized in that: include: A paclitaxel twin-drug compound as claimed in claim 1 or 2 is mixed with albumin in a composite solution to prepare paclitaxel twin-drug nanoparticles.

15. The method for preparing paclitaxel twin-drug nanoparticles according to claim 14, characterized in that: The following steps are involved: S1, dissolving the paclitaxel twin drug compound in a mixture of organic solvents to prepare a drug solution; S2, preparing an albumin aqueous solution as an albumin solution; S3, adding the drug solution into the albumin solution and preparing a nanosuspension by ultrasound; S4, evaporating and removing the residual organic solvent in the suspension obtained in step S3; S5, concentrating the nanosuspension by ultrafiltration to obtain albumin-paclitaxel twin-drug nanoparticles.

16. The method for preparing paclitaxel twin-drug nanoparticles according to claim 10, characterized in that: The organic solvent in step S1 includes any one or more of dichloromethane, chloroform, ethanol, DMSO, DMF, and acetonitrile; the concentration of albumin in the albumin solution in step S2 is 1-100 mg / mL.

17. Use of the paclitaxel twin-drug compound of claim 1 in the preparation of a drug for treating cancer, wherein the cancer includes non-small cell lung cancer, pancreatic cancer, cervical cancer, ovarian cancer, bladder cancer, breast cancer, lung cancer, gastric cancer and melanoma.

18. The use according to claim 17, characterized in that The paclitaxel twin drug compound includes any one drug molecule selected from T785, di-ABZI, SR07, Crizotinb, Palbociclib, MK1775, MK2206, AZD7762, Ceritinib, Navitoclax, Dasatinib and Exatecan.

19. Use of the paclitaxel twin-drug nanoparticles according to claim 7 in the preparation of a drug for treating cancer, wherein the cancer includes non-small cell lung cancer, pancreatic cancer, cervical cancer, ovarian cancer, bladder cancer, breast cancer, lung cancer, gastric cancer and melanoma.

20. The use according to claim 19, characterized in that The paclitaxel twin-drug nanoparticles also include albumin, and the paclitaxel twin-drug compound is bonded to the albumin; the albumin includes at least one of bovine serum albumin, human serum albumin, ovalbumin and recombinant human serum albumin; the bonding is a non-covalent bond; the paclitaxel twin-drug nanoparticles include at least one paclitaxel twin-drug compound.

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