Search

CN-122005789-A - Pancreatic cancer targeting and electric field response nano micelle and preparation method and application thereof

CN122005789ACN 122005789 ACN122005789 ACN 122005789ACN-122005789-A

Abstract

The invention provides a pancreatic cancer targeting and electric field response nano micelle, a preparation method and application thereof, wherein the nano micelle is subjected to blood circulation after intravenous injection and is combined to a pancreatic cancer lesion site through the specificity of surface-modified pancreatic cancer targeting peptide and P-lectin1 highly expressed on the surface of pancreatic cancer cells. The electroresponse group ferrocene in the nano micelle has hydrophobicity in a reduced state and forms a part of the nano material. After the pulse electric field is ablated, the nano-material is oxidized to be hydrophilic, so that the nano-material structure is broken, and the CpG ODN of the immune adjuvant is released. Pulsed electric field ablation-induced cell death releases TAA and DAMP, promoting APC maturation, M1 macrophage polarization, pro-inflammatory cytokine secretion, and CTL activation. This synergy maintains the immunostimulation, prolonging the transient immunomodulation effects of pulsed electric field ablation. The nano micelle has wide application prospect in improving the pancreatic cancer pulse electric field ablation curative effect.

Inventors

  • Jiang tianan
  • XIE LITING
  • ZHANG CHENGYUE

Assignees

  • 浙江大学医学院附属第一医院(浙江省第一医院)

Dates

Publication Date
20260512
Application Date
20260206

Claims (7)

  1. 1. A pancreatic cancer targeting and electric field response nano micelle is characterized by comprising, by mass, 75% -92% of PLA2K-PEG2K, KTLLPTP% -10%, 1% -5% of Fc, 1% -5% of CpG ODN and 1% -5% of OA-SPIO.
  2. 2. The nano-micelle according to claim 1, wherein the nano-micelle has a particle size of 140-160 nm and a dispersibility index of 0.10-0.15.
  3. 3. A method of preparing a nanomicelle according to any of claims 1-2, characterized in that it comprises the steps of: s1, weighing PLGA2K-PEG2K-NHS, KTLLPTP and triethylamine, dissolving in 3mL of DMF, stirring and reacting at room temperature for 12 h with the mass ratio of PLGA2K-PEG2K-NHS to KTLLPTP to triethylamine being 1:1.1:3, dialyzing in pure water for 24 h, collecting dialyzate, and freeze-drying to obtain PLA2K-PEG2K-KTLLPTP; S2, respectively weighing PEG2K-PLA2K, DCC, ferrocene formate and DMAP with the mass ratio of 10:3:6:5, dissolving in 20mL of dichloromethane CH 2 Cl 2 , magnetically stirring and reacting for 24 hours at room temperature, dialyzing for 24 hours in pure water for h, collecting dialysate, and freeze-drying to obtain PLA2K-PEG2K-Fc; S3, dissolving PLA2K-PEG2K-KTLLPTP, PLA2K-PEG2K and PLA2K-PEG2K-Mal obtained in the step S1, wherein the mass ratio of PLA2K-PEG2K-Fc obtained in the step S2 is (5-10) (0-5) (1-5) in 2mL of mixed solution, and adding 40-200 mu L of OA-Fe 3 O 4 with the concentration of 5 mg/mL; s4, carrying out ultrasonic treatment on the mixed solution obtained in the step S3 for 1min, and slowly dripping 2 mL of water in the ultrasonic treatment process to obtain emulsion; s5, adding 0.2-1mg of CpG ODN-SH into the emulsion obtained in the step S4, dropwise adding 0.5M NaHCO 3 solution to adjust the pH of the liquid to 7.5-8.0, stirring for 4 hours, dialyzing in pure water for 48: 48 h, collecting dialysate, and freeze-drying to obtain the nano micelle.
  4. 4. The method of claim 3, wherein the dialysis bags used in the dialysis process in steps S1 and S2 have a molecular weight cut-off of 2000 Da.
  5. 5. The method according to claim 3, wherein the mixed solution in step S3 is formed by mixing tetrahydrofuran and dimethyl sulfoxide according to a volume ratio of 9:1.
  6. 6. The method according to claim 3, wherein the dialysis bag has a molecular weight cut-off of 7000 Da during the dialysis in step S5.
  7. 7. Use of the nanomicelle of any of claims 1-2 in the manufacture of a pancreatic cancer targeted pulsed electric field ablative drug.

Description

Pancreatic cancer targeting and electric field response nano micelle and preparation method and application thereof Technical Field The invention belongs to the technical field of biological medicines, and particularly relates to a pancreatic cancer targeting and electric field response nano micelle and a preparation method and application thereof. Background Pancreatic Ductal Adenocarcinoma (PDAC) remains the fourth leading cause of cancer-related death worldwide, with 5-year survival rates below 7%. Pulsed electric field ablation (PFA) is a minimally invasive ablation procedure that uses a transient high voltage microsecond pulsed electric field (μpef) to trigger irreversible cell membrane permeabilization, resulting in cancer cell death. Compared with other local ablation technologies, the PFA has the remarkable and unique advantages that important structures such as a vasculature system, a pancreatic duct and nerves can be effectively reserved, interference of a heat sink effect can be eliminated, and tumor tissues positioned near the key structures can be safely and accurately ablated. Recent preclinical studies have shown that PFA ablation can elicit anti-tumor immunity. However, the Tumor Microenvironment (TME) of PDACs exhibits immune "cold" features, manifested by insufficient infiltration of immune cells, defective antigen presentation, and constant accumulation of immunosuppressive molecules. This TME drives tumor cells to escape immunologically, which in turn leads to rapid tumor recurrence following PFA treatment. In order to enhance antitumor efficacy, immune reactivation is critical, while immunoadjuvants are a promising strategy. Among various immunoadjuvants, cytosine-phosphate-guanine (CpG) Oligodeoxynucleotides (ODNs) are widely considered as one of the most potent stimulators. They activate Toll-like receptor 9 (TLR 9) -mediated signaling pathways, leading to upregulation of pro-inflammatory cytokines and chemokines. In addition, cpG ODNs promote maturation of Antigen Presenting Cells (APCs) and enhance Th1 immune responses. Thus, combining PFA with immunotherapy (electric immunotherapy) may result in sustained efficacy and wider applicability, including restriction of tumor metastasis, etc. However, the clinical use of CpG ODNs is limited by a number of challenges that are susceptible to nuclease degradation, maldistribution, and systemic toxicity, which severely compromises their immunostimulatory efficacy. To address these limitations, nanomedicine, by virtue of its unique advantages, builds a very promising strategy framework for the synergistic co-delivery of multiple therapeutic agents. The strategy not only can skillfully avoid the obstruction of various biological barriers, but also can obviously enhance the targeting delivery efficiency and accuracy of the therapeutic agent to pathological positions. Researchers have developed a variety of responsive nanomaterials, such as nanoliposomes and micelles that are sensitive to TME (e.g., pH), light or temperature, which in part enhance intratumoral drug accumulation. However, endogenous stimuli present in both tumor microenvironment and normal physiological environments may lead to accidental release of drugs in healthy tissue, compromising the specificity of TME-responsive nanomaterials. Furthermore, due to the poor tissue penetration depth of light, the light responsive nanomaterial is more suitable for superficial tumors than for deep tumors such as PDACs. In PFA treatment, the electric field can cover the pancreatic cancer tumor area, and the electric field is used as external energy and can be used as a stimulus source to activate drug release. At present, no nano micelle material which can simultaneously solve pancreatic cancer targeting and PFA electric field responsiveness and is safe and simple to prepare is available. References are referred to as follows: 1、Tempero, M.A., Malafa, M.P., Al-Hawary, M., Behrman, S.W., Benson, A.B., Cardin, D.B., Chiorean, E.G., Chung, V., Czito, B., Del Chiaro, M., et al. (2021). Pancreatic Adenocarcinoma, Version 2.2021, NCCN Clinical Practice Guidelines in Oncology. Journal of the National Comprehensive Cancer Network : JNCCN 19, 439-457. doi:10.6004/jnccn.2021.0017. 2、Burbach, B.J., O'Flanagan, S.D., Shao, Q., Young, K.M., Slaughter, J.R., Rollins, M.R., Street, T.J.L., Granger, V.E., Beura, L.K., Azarin, S.M., et al. (2021). Irreversible electroporation augments checkpoint immunotherapy in prostate cancer and promotes tumor antigen-specific tissue-resident memory CD8+ T cells. Nature communications 12, 3862. doi:10.1038/s41467-021-24132-6. 3、Zhu, Y.S., Tang, K., and Lv, J. (2021). Peptide-drug conjugate-based novel molecular drug delivery system in cancer. Trends in pharmacological sciences 42, 857-869. doi:10.1016/j.tips.2021.07.001. 4、Fatima, M., Almalki, W.H., Khan, T., Sahebkar, A., and Kesharwani, P. (2024). Harnessing the Power of Stimuli-Responsive Nanoparticles as an Effective Ther