Search

CN-121971390-A - Mn coating based on tumor cell membrane and liposome2+Immune nano-drug modified by graphene quantum dots and preparation method thereof

CN121971390ACN 121971390 ACN121971390 ACN 121971390ACN-121971390-A

Abstract

The invention discloses a photothermal immune nano-drug based on a tumor cell membrane and liposome coated manganese ion modified graphene quantum dot and a preparation method thereof, and belongs to the field of nano-drugs. The preparation method comprises the steps of (1) preparing 4T1 cell membrane tm and liposome Lip, (2) preparing a hybrid nano drug GMP loaded with manganese ions and mPEG-NH 2 , and (3) preparing GMP@liptm. According to the invention, metabolizable graphene quantum dots are selected as the photothermal treatment nano platform, so that the preparation method has higher drug loading rate and photothermal conversion rate, and can effectively scald tumor cells. The encapsulation of mPEG-NH 2 , lip and tm can improve the tumor targeting of the nano-drug, and Mn 2+ activates the cgAs-STING signal channel so as to excite the immune response of the organism. According to the invention, the tumor cell membrane and the liposome are used for wrapping Mn 2+ modified graphene quantum dots for the first time, so that the synergistic treatment effect of photo-thermal ablation-immune activation-targeted delivery of breast cancer is realized, the design method is simple, the reaction condition is mild, and a new strategy is provided for the accurate treatment of breast cancer.

Inventors

  • ZHOU BENQING
  • GAN YINGYING

Assignees

  • 汕头大学

Dates

Publication Date
20260505
Application Date
20260123

Claims (10)

  1. 1. The immune nano-drug is characterized by comprising a carrier and a load, wherein the carrier wraps the load, and the load comprises manganese modified quantum dots.
  2. 2. The immune nanodrug of claim 1, wherein the carrier comprises one or more of a cell membrane, a liposome membrane.
  3. 3. The immune nanodrug according to claim 2, wherein the carrier comprises a bilayer membrane structure formed by the encapsulation of the liposome membrane by the cell membrane, wherein the cell membrane comprises a tumor cell membrane.
  4. 4. The immune nano-drug according to claim 1, wherein the carrier comprises mPEG-NH 2 、Mn 2+ co-modified quantum dots, wherein the quantum dots comprise graphene quantum dots.
  5. 5. A method of preparing an immune nano-drug according to claim 2, comprising: A. dispersing massive quantum dot powder into ultrapure water to obtain a dispersion liquid of the quantum dots; B. Modifying the quantum dots to obtain a GMP aqueous solution; C. preparing the liposome membrane; D. Adding the GMP aqueous solution into the liposome membrane, and hydrating and extruding to obtain a single-layer membrane-coated GMP aqueous solution; E. extracting the cell membrane from the cell; F. And adding the cell membrane into an aqueous solution of GMP wrapped by the single-layer membrane, and extruding and purifying to obtain the immune nano-drug.
  6. 6. The method according to claim 5, comprising: A. Dispersing massive quantum dot powder in ultrapure water, and performing ultrasonic dispersion to obtain a dispersion liquid of the quantum dots; B. Uniformly mixing the aqueous dispersion of the quantum dots with MnCl 2 aqueous solution, then adding the mPEG-NH 2 aqueous solution, uniformly mixing, and dialyzing to obtain the GMP aqueous solution; C. Dispersing lecithin, cholesterol and DSPE-mPEG 2000 in absolute ethyl alcohol, and performing rotary evaporation to obtain the liposome membrane; D. adding the liposome membrane into the GMP aqueous solution, and obtaining the single-layer membrane-coated GMP aqueous solution through hydration and extrusion; E. Extracting the cell membrane from the cells cultured in vitro; F. Adding the cell membrane into the aqueous solution of GMP wrapped by the monolayer membrane, extruding and purifying to obtain the immune nano-drug.
  7. 7. The method according to claim 6, wherein the mass ratio of the quantum dots to Mn 2 + in the MnCl 2 aqueous solution, to mPEG-NH 2 in the mPEG-NH 2 aqueous solution, to the liposome membrane, to the cell membrane comprises 1:14:5:10:5, wherein the size of the quantum dots in step A comprises 6-10 nm, the concentration of the dispersion of the quantum dots comprises 0.2-2 mg/mL, wherein the concentration of Mn 2+ in the MnCl 2 aqueous solution comprises 50-500 mM, wherein the concentration of mPEG-NH 2 aqueous solution comprises 1-10 mg/mL, wherein the molecular weight of mPEG-NH 2 in the mPEG-NH 2 aqueous solution comprises 2000, wherein the molecular weight of the dialyzed entrapped membrane comprises 3500 Da, wherein the mass-volume ratio of the lecithin, the cholesterol, the DSPE-mPEG 2000 , and the absolute ethanol in step C comprises 2~20 mg:0.5~5 mg:0.15~1.5 mg:2~5 mL, wherein the concentration of Mn 2+ in the MnCl 2 aqueous solution comprises 1-10 mg/mL, and wherein the number of cells in vitro cultured in the step E comprises 1X 10 cells.
  8. 8. The method according to claim 5, comprising: A. dispersing massive quantum dot powder in ultrapure water, carrying out ultrasonic dispersion for 40 min by using an ultrasonic cell grinder ice bath of a No. 6 amplitude transformer under the condition that the power is 600W and the ultrasonic frequency is 1.0 s and 2.0 s, and standing for standby at 4 ℃ in a dark place; B. Weighing according to the required mass of Mn 2+ converted into the mass of corresponding MnCl 2 , preparing a MnCl 2 aqueous solution, mixing the dispersion liquid of the quantum dots with the MnCl 2 aqueous solution in a light-proof environment, then carrying out ultrasonic treatment on the mixture under the power conditions of 15-25 ℃ and 300W of an ultrasonic cleaning machine for 30 min, stirring the mixture for 12 h, adding the mPEG-NH 2 aqueous solution, carrying out ultrasonic treatment on the mixture under the power conditions of 15-25 ℃ and 300W of the ultrasonic cleaning machine for 30 min, stirring the mixture for 12 h, and finally carrying out dialysis on the mixture for 24 h by using ultrapure water to obtain the GMP aqueous solution; C. Dispersing the lecithin, the cholesterol and the DSPE-mPEG 2000 in the absolute ethyl alcohol, and spin-steaming the mixture into a uniform film at the temperature of 40 ℃ and the rotating speed of 80 rpm by a spin-steaming instrument to obtain the liposome film; D. Adding the liposome membrane into the GMP aqueous solution, setting the water bath temperature to be 40-60 ℃ for hydration of 1h, and extruding the mixture through a 200 nm filter membrane of a liposome extruder for 11 times to obtain the single-layer membrane-coated GMP aqueous solution; E. Extracting the cell membrane from the cells cultured in vitro using a cell membrane extraction kit; F. adding the cell membrane into the GMP aqueous solution wrapped by the monolayer membrane, uniformly mixing, extruding through a 400 nm filter membrane for 11 times, and ultrafiltering for three times under the conditions of an ultrafiltration tube 8000 rpm with the molecular weight cutoff of 100 KDa and 4 ℃ to obtain the immune nano-drug.
  9. 9. Use of a nano-drug according to claim 1 for the preparation of a targeted drug for photothermal immune complex treatment of cancer.
  10. 10. The use of claim 9, wherein the cancer comprises breast cancer.

Description

Immune nano-drug based on tumor cell membrane and liposome coated Mn 2+ modified graphene quantum dots and preparation method thereof Technical Field The invention relates to the field of nano-drugs, in particular to an immune nano-drug based on a tumor cell membrane and a liposome coated Mn 2+ modified graphene quantum dot and a preparation method thereof. Background Triple negative breast cancer is a common malignant tumor threatening the health of women, and is a serious difficulty in the current treatment field due to the characteristics of high invasiveness, high recurrence rate and high metastasis rate, and along with the continuous progress of medical research, the diagnosis and early treatment of the triple negative breast cancer have certain achievements. Advanced breast cancer remains a significant challenge that humans must face. The traditional treatment means such as surgical excision, chemotherapy, radiotherapy and the like still have the problems of poor targeting, strong toxicity and side effects, easy initiation of immunosuppressive Tumor Microenvironment (TME) and the like. The defects of the prior art are that the current main clinical treatment means plays a role in a certain stage, but the defects that 1, the targeting property of the medicine is poor, the toxicity is strong, and the medicine with poor targeting property is easy to accumulate in normal tissues to cause serious toxic and side effects, so that the treatment effect is poor are all existed. 2. The single treatment means is that separate chemotherapy or radiotherapy can aggravate the hypoxia, acidity characteristic and immunosuppression state of the tumor microenvironment, thereby reducing the self-healing effect, inhibiting the immunocyte activity and finally causing the treatment recurrence and metastasis. 3. The material problems are that most of the existing drug carrier materials are non-degradable materials, and can cause biotoxicity or long-term in-vivo residual problems. Aiming at the limitations of traditional treatment, immunotherapy becomes an important breakthrough in the field of breast cancer treatment. The core advantage of the method is that the method can activate systemic anti-tumor immune response of organisms, promote infiltration and activity of immune cells by regulating immune suppression microenvironment (such as activating cGAS-STING signal channels), and radically reduce recurrence and metastasis risks. However, the single immunotherapy has a problem of low response rate to advanced local solid tumors, so that a comprehensive therapeutic strategy is urgently needed, and the immunotherapy is combined with other therapeutic means to overcome the limitations of the existing therapeutic means and realize synergistic effect. The ideal therapeutic scheme should eliminate local solid tumor rapidly through combined therapy, induce tumor specific antigen through immunotherapy, activate organism immune response, thus more accurately identify and eliminate the residual tumor cells, and prevent tumor recurrence. Photothermal therapy (Photothermal Therapy, PTT) has shown unique advantages as a novel topical treatment in a variety of solid tumor treatments. The principle is that the near infrared light irradiates a specific photo-thermal agent to efficiently convert light energy into heat energy, so that the tumor cells are killed locally and accurately. The therapy has the advantages of minimally invasive property, space-time controllability and high tumor killing efficiency, can accurately focus on focuses and reduce damage to surrounding normal tissues, and simultaneously, the photothermal therapy can induce ICD, release tumor-associated antigens and damage-associated molecular patterns (DAMPs), activate antigen presenting cells, enhance tumor immunogenicity, provide a 'target spot' for immunotherapy, can generate remarkable synergistic effect in combination with the immunotherapy, and finally realize more efficient and lasting anti-tumor effect. In recent years, the synergistic combination of photothermal therapy and immunotherapy has brought up a brand new development opportunity by virtue of the unique small-size effect and the convenient functional modification advantages of the nano material. The nano material can not only rely on the small-size characteristic to accurately gather at the local part of the tumor to strengthen the killing efficiency of the photothermal treatment on tumor cells, but also can activate the anti-tumor immune response of the organism by surface modification of an immune adjuvant, thereby remarkably improving the overall effect of the tumor treatment. Among them, graphene Quantum Dots (GQDs) are ideal photothermal agents due to excellent optical stability, gao Guangre conversion efficiency and good biocompatibility. However, purely hybrid GQDs are readily cleared by the in vivo single-core-macrophage system. The tumor cell membrane is utilized to endow the nano-carrier with the f