CN-122005847-A - Targeting SELENOP+Macrophage liposome, preparation method thereof and application thereof in lung squamous carcinoma treatment

Abstract

The invention relates to the technical field of biological medicines, and particularly discloses a liposome targeting SELENOP + macrophages, a preparation method thereof and application thereof in lung squamous cell carcinoma treatment. The liposome takes SELENOP antibody as a targeting modification molecule, loads si-SELENOP and si-ABCA1/G1, and realizes the accurate delivery of siRNA in target cells through the specific combination of the antibody and SELENOP receptors on the surface of SELENOP + macrophages. The targeted liposome prepared by the invention can specifically inhibit the cholesterol efflux pathway of SELENOP + macrophages, reverse the polarization of the pro-tumor phenotype, remodel the tumor immune microenvironment and restore the anti-tumor activity of immune cells such as CD8 + T cells, NK cells and the like. When the liposome is used alone or in combination with PD-1 antibody, the liposome can obviously inhibit the growth and metastasis of lung squamous carcinoma, improve the treatment effect, provide a brand-new 'metabolism-immunity' synergistic targeting treatment strategy for lung squamous carcinoma, and have good clinical application prospect.

Inventors

• CHEN XIANGUO
• PAN KAILING
• CHEN XING

Assignees

• 浙江嘉臣生物科技有限公司

Dates

Publication Date

20260512

Application Date

20260206

Claims (6)

1. 1. The liposome targeting SELENOP + macrophages is characterized by comprising a liposome carrier, a targeting modification molecule and a loaded functional nucleic acid, wherein the targeting modification molecule is a SELENOP antibody, the functional nucleic acid is a combination of si-SELENOP and si-ABCA1/G1, and the liposome carrier consists of ionizable lipid MC3, cholesterol and neutral phospholipid DOPE according to a molar ratio of 2:1:1.
2. 2. The liposome targeting SELENOP + macrophages of claim 1, wherein the molar ratio of si-SELENOP to si-ABCA1/G1 is 1:1, the total drug loading of siRNA in the liposome is 5-10 μg/mL, and the mass ratio of SELENOP antibody to liposome carrier is 1:10-1:20.
3. 3. The preparation method of the targeted SELENOP + macrophage liposome according to claim 1 or 2 is characterized by comprising the following steps of S1, adopting a thin film method to prepare a blank liposome composed of MC3, cholesterol and DOPE, S2, mixing si-SELENOP with si-ABCA1/G1, adding the blank liposome, incubating at room temperature for 30-60min to obtain siRNA-loaded liposome, S3, covalently connecting SELENOP antibody to the surface of the siRNA-loaded liposome by an EDC/NHS activation method, and dialyzing and purifying to obtain the target liposome.
4. 4. The preparation method according to claim 3, wherein the film preparation conditions in the step S1 are 37 ℃, the rotation speed is 100rpm, the reaction is carried out for 30-60min, the film is dried in vacuum for 4-6h, then ultrapure water is added for hydration, ice bath ultrasonic treatment and then the film is extruded through a 0.45 μm filter membrane, the incubation conditions in the step S3 are 4 ℃ and are incubated for 12-24h in a dark place, a 10kDa dialysis bag is adopted for dialysis, and the film is dialyzed for 24-48h in PBS buffer solution with pH of 7.4.
5. 5. Use of the targeted SELENOP + macrophage liposome according to claim 1 or 2 in the preparation of a medicament for treating lung squamous cell carcinoma.
6. 6. The use according to claim 5, wherein the therapeutic agent is the liposome alone or in combination with the PD-1 antibody in a mass ratio of 5:1-10:1, by intravenous injection at a dose of 1-5mg per kg body weight and at a frequency of 1-2 times per week.

Description

Liposome targeting SELENOP + macrophages, preparation method thereof and application thereof in lung squamous carcinoma treatment Technical Field The invention relates to the technical field of biological medicines, in particular to a tumor targeting drug delivery system, a gene therapy and tumor immunotherapy crossing technology, and especially relates to a functional liposome taking SELENOP + macrophages as targets, a preparation method thereof and application of the liposome in accurate treatment of lung squamous cell carcinoma. Background Lung squamous cell carcinoma (Lung Squamous Cell Carcinoma, LUSC) is one of the main subtypes of lung cancer, accounting for 30% -50% of primary lung cancer in China, and the annual incidence rate is about 35/10 ten thousand. The pathogenesis of the traditional Chinese medicine is complex, is closely related to long-term smoking, environmental pollutant exposure, chronic lung inflammation, genetic factors and the like, and has the characteristics of strong invasiveness, early transfer and poor prognosis. Along with the progress of the disease, the patients can have symptoms such as cough, hemoptysis, chest pain, dyspnea and the like, and can also have distant metastasis in late stage, thereby seriously threatening the life health of the patients and bringing heavy medical burden to families and society. However, existing lung squamous cancer treatment regimens suffer from the following drawbacks: 1. Partial clinical treatment relies on platinum chemotherapy combined with PD-1 inhibitors, and over 70% of patients can have primary or secondary drug resistance, and the positive rate of lung squamous carcinoma driving gene mutation (such as EGFR/ALK) is less than 5%, and targeted therapeutic drugs are lacked; 2. Traditional chemotherapeutics have poor targeting property, damage normal tissues easily while killing tumor cells, and cause serious systemic side effects; The response rate of PD-1 inhibitor single drug treatment to immunosuppressive lung squamous carcinoma is low, and immune escape barrier mediated by lipid metabolism abnormality is difficult to break through; 4. conventional drug delivery systems cannot achieve accurate drug delivery, further limiting therapeutic efficacy enhancement. Disclosure of Invention The invention aims to solve the defects of insufficient targeting, limited treatment effect, difficult reversion of immunosuppressive microenvironment and the like of lung squamous carcinoma treatment in the prior art, and provides a liposome targeting SELENOP + macrophages, a preparation method and application thereof. In order to achieve the aim, the technical scheme is that the liposome targeting SELENOP + macrophages comprises a liposome carrier, a targeting modification molecule and loaded functional nucleic acid, wherein the targeting modification molecule is a SELENOP antibody, the functional nucleic acid is a combination of si-SELENOP and si-ABCA1/G1, and the liposome carrier consists of ionizable lipid MC3, cholesterol and neutral phospholipid DOPE. Further, the molar ratio of the ionizable lipid, cholesterol, and neutral phospholipid is 2:1:1. The adoption of the further scheme has the beneficial effects that the structural stability and the siRNA entrapment efficiency of the liposome are guaranteed by the accurate proportioning, and a foundation is laid for the subsequent targeted drug delivery and gene silencing effects. Further, the molar ratio of si-SELENOP to si-ABCA1/G1 is 1:1, and the total drug loading of siRNA in the liposome is 5-10 mug/mL. The beneficial effect of the adoption of the further scheme is that the double siRNA synergistic effect can simultaneously block the expression and the channel function of the target gene, the drug loading rate gives consideration to the treatment effect and the stability of the liposome, and the liposome aggregation or the influence on the curative effect caused by the too high drug loading rate is avoided. Further, the mass ratio of SELENOP antibodies to liposome carrier is 1:10-1:20. The further scheme has the beneficial effects that the modification proportion of the antibody is reasonably controlled, the specific recognition capability of the liposome on SELENOP + macrophages is ensured, and meanwhile, the abnormal structure of the liposome caused by excessive antibody is avoided. Furthermore, the application of the targeted SELENOP + macrophage liposome in preparing lung squamous cancer therapeutic drugs is characterized in that the therapeutic drugs are compound preparations consisting of the liposome singly or with PD-1 antibody according to the mass ratio of 5:1-10:1, the administration mode is intravenous injection, the administration dosage is 1-5mg per kilogram of body weight, and the administration frequency is 1-2 times per week. The adoption of the further scheme has the advantages that the clinical application scene and the use mode are clear, the accurate targeting therapeutic effect