CN-121971405-A - Composite taxol albumin nanoparticle as well as preparation method and application thereof

CN121971405ACN 121971405 ACN121971405 ACN 121971405ACN-121971405-A

Abstract

The invention discloses a composite taxol albumin nanoparticle, a preparation method and application thereof, wherein human serum albumin is used for co-loading taxol and a non-nucleotide interferon gene stimulation factor agonist in a simple mode to form nanoparticles with uniform size, proper particle size, better dispersibility and stable physicochemical property, the added non-nucleotide interferon gene stimulation factor agonist forms a dimer in an acidic tumor microenvironment, and the active targeting of a carrier is combined to enable the nanoparticles to have excellent selectivity on tumor tissues, and the non-nucleotide interferon gene stimulation factor agonist is used for starting innate immunity and combining with adaptive immunity of taxol to solve the defect of low immune response of cold tumors. The nano-particles provided by the invention can solve the problem of strong toxic and side effects of taxol and simultaneously synergistically enhance the anti-tumor immunotherapy, thereby achieving the effect of combined therapy.

Inventors

WEI HUA
CAO ZHEN
YU CUIYUN
CHEN MINXUAN
CHEN YONGCHENG
HU JIANWEI
HUANG YUN
ZHANG QIN

Assignees

南岳生物制药有限公司

Dates

Publication Date: 20260505
Application Date: 20260209

Claims (10)

1. A preparation method of a composite taxol albumin nanoparticle is characterized in that taxol and a non-nucleotide interferon gene stimulation factor agonist are dissolved in an organic solvent to be used as an organic phase, an aqueous solution containing human serum albumin HSA is used as an aqueous phase, the organic phase is injected into the aqueous phase drop by drop under a high-speed shearing condition to form a primary emulsion, and the primary emulsion is homogenized under high pressure to remove the organic solvent to obtain the composite taxol albumin nanoparticle.
2. The method for preparing the composite paclitaxel albumin nanoparticle according to claim 1, comprising the following steps: Step one, weighing paclitaxel and a non-nucleotide interferon gene stimulating factor agonist, dissolving the paclitaxel and the non-nucleotide interferon gene stimulating factor agonist in an organic solvent to serve as an organic phase, and taking an aqueous solution containing human serum albumin HSA as an aqueous phase; step two, the organic phase is injected into the water phase drop by drop under the condition of high-speed shearing to form primary emulsion; Step three, placing the primary emulsion in a high-pressure homogenizer, circulating for a plurality of times under preset homogenizing pressure to obtain a nanometer suspension of the paclitaxel and the non-nucleotide interferon gene stimulating factor agonist, and removing the organic solvent by rotary steaming at preset temperature and pressure for preset time to obtain a composite paclitaxel albumin nanoparticle suspension aqueous solution; and step four, freeze-drying the mixed suspension aqueous solution of the composite paclitaxel albumin nanoparticle by a freeze dryer to obtain the composite paclitaxel albumin nanoparticle.
3. The method for preparing composite paclitaxel albumin nanoparticle according to claim 2, wherein the non-nucleotide interferon gene stimulating factor agonist in the first step is 2-methoxycarbonyl benzyl sulfonic acid.
4. The method for preparing composite paclitaxel albumin nanoparticle according to claim 3, wherein in the first step, the concentration of paclitaxel in the organic phase is 5-10mg/mL, the molar ratio of paclitaxel to 2-methoxycarbonylbenzyl sulfonic acid is 1:1-5, and the organic solvent is chloroform or a mixed solvent of chloroform and ethanol; The concentration of the human serum albumin HSA in the aqueous phase is 0.5-30mg/mL/, the aqueous solution is ultrapure water or sodium chloride aqueous solution, and the volume ratio of the aqueous phase to the organic phase is 40:1-60:1.
5. The method for preparing the composite paclitaxel albumin nanoparticle according to claim 2, wherein in the second step, the high-speed shearing condition is that after the organic phase is added into the water phase under 5000-7000 revolutions of a high-speed shearing emulsifying machine, the shearing is continued for 5min to form a primary emulsion; in the third step, the homogenizing pressure of the high-pressure homogenizer is 800-1600bar, and the cycle times are 3-7 times.
6. The method for preparing the composite taxol albumin nanoparticle according to claim 4, wherein the concentration of taxol in the organic phase is 10mg/mL, the feeding mole ratio of taxol to 2-methoxycarbonyl benzyl sulfonic acid is 1..1, the organic solvent is chloroform, the aqueous solution is ultrapure water, the volume ratio of the aqueous phase to the organic phase is 50..1, the concentration of human serum albumin HSA in the aqueous phase is 5mg/mL, the homogenizing pressure of the high-pressure homogenizer in the third step is 1200bar, and the cycle number is 5.
7. The method for preparing the composite paclitaxel albumin nanoparticle according to claim 2, wherein in the third step, the spin-steaming pressure is 81.064 kPa, the spin-steaming time is 10-30min, and the spin-steaming temperature is 35 ° C-40 ° ℃; And (3) taking out the nanoparticle suspension aqueous solution obtained in the step four after pre-freezing in a low-temperature refrigerator, and freeze-drying in a freeze dryer to obtain the composite taxol albumin nanoparticle freeze-dried powder.
8. The composite paclitaxel albumin nanoparticle is characterized by being prepared by adopting the preparation method of the composite paclitaxel albumin nanoparticle in any one of claims 1-8.
9. The use of the composite paclitaxel albumin nanoparticle according to claim 8, wherein the composite paclitaxel albumin nanoparticle is used for preparing a drug for treating triple negative breast cancer.
10. The use of the composite paclitaxel albumin nanoparticle according to claim 9, wherein the composite paclitaxel albumin nanoparticle is used for preparing a medicament for activating STING pathway and enhancing expression of phosphorylation protein related to STING pathway, thereby activating innate immunity of tumor cells and inducing generation of strong immune memory.

Description

Composite taxol albumin nanoparticle as well as preparation method and application thereof Technical Field The invention belongs to the technical field of medicine preparation, and in particular relates to a composite taxol albumin nanoparticle and application thereof in synergistic chemo-immunotherapy of triple negative breast cancer Background Cancer treatment has made tremendous progress over the past decades. Among the numerous therapeutic approaches, chemotherapy is currently the mainstream therapeutic strategy due to the high cytotoxicity of drugs to tumor cells, however, conventional chemotherapeutic drugs such as paclitaxel always exhibit inherent limitations such as nonspecific distribution, high toxicity, poor bioavailability, rapid blood clearance, poor solubility in physiological environments, etc., which seriously affect the clinical transformation of drugs. Human serum albumin is one of the most abundant proteins in blood, has a molecular weight of 66.5 kDa, has a biological half-life of 19d in vivo, and plays an important physiological role in maintaining the osmotic pressure of plasma. In addition, tumor cells express albumin receptors, which can combine various hydrophobic and hydrophilic therapies to form stable complexes with high drug loading. Human Serum Albumin (HSA) is widely used for the delivery of various therapeutic drugs such as small molecule chemotherapeutics, polypeptides, proteins, nucleic acids, etc., due to its high solubility, stability to pH changes (pH between 4-9), good biocompatibility, biodegradability, low toxicity and non-immunogenic properties. In 2005, abraxane (albumin-bound paclitaxel) developed by the american celer gene company is approved by the FDA for marketing, and has become a major drug for various cancers such as breast cancer. Due to abnormal metabolism of tumors and complex microenvironments, the efficacy of chemotherapy alone is often poor. The development of multi-drug combination therapy is expected to bring new hopes for patients. At present, the combination of Abraxane, carboplatin, gemcitabine and other antitumor drugs is widely used clinically, and the survival rate of patients can be improved by combined treatment, but most of the current clinical combined treatment is a chemotherapeutic drug, and the problems of low bioavailability, poor targeting, large toxic and side effects, repeated drug action and the like generally exist. Compared with the traditional cancer treatment method, the immunotherapy has the advantages of improving the fight capability of the immune system in the human body by mobilizing immune cells capable of recognizing tumors in the human body, indirectly killing and controlling cancers by virtue of the immune cells, along with small side effect, safety and effectiveness. However, due to the nature of triple negative breast cancers, immunotherapy in most patients fails to achieve the expected clinical therapeutic effect, mainly because the effectiveness of tumor immunotherapy is limited by the low immunogenicity of the tumor and immunosuppressive microenvironment. Thus, there is a need to overcome the existing bottleneck of tumor therapy while overcoming the high toxic side effects of chemotherapy and targeted delivery of immunotherapy. The invention patent CN109806241A discloses an albumin-entrapped double-drug nanoparticle and a preparation process thereof. The preparation process of the dual-drug albumin nanoparticle provided by the scheme verifies the advantages of the taxol/resveratrol albumin nanoparticle in-vitro anticancer activity through MTT experiments. However, the proposal mainly discloses a preparation process of the albumin-entrapped paclitaxel and the resveratrol, and does not clarify a combination mechanism, and the proposal is mainly applied to the chemotherapy combination treatment of drug-resistant cancers. The invention has the main application fields of chemotherapy and immunotherapy combined treatment, and the embodiment illustrates the mechanism of the combined treatment, thereby providing detailed data support for clinical medicine experiments. Disclosure of Invention The invention aims to solve the problems of low responsiveness of tumor immunotherapy and accurate delivery of non-nucleotide interferon gene stimulus factors, and provides a composite taxol albumin nanoparticle, a preparation method and application thereof. In order to reduce toxic and side effects of PTX and enhance curative effect, the invention selects FDA approved non-nucleotide interferon gene stimulating factor agonist and taxol to prepare a composite nanoparticle and is used for chemotherapy combined immunotherapy of cancer. The nanoparticles with proper particle size are prepared by combining paclitaxel and non-nucleotide interferon gene stimulators, so that a better tumor targeting effect is achieved, the toxic and side effects of paclitaxel are reduced, and the anti-tumor curative effect is enhanced through immune activatio