CN-121971651-A - Metal-based nanoparticle loaded with traditional Chinese medicine and preparation method and application thereof

CN121971651ACN 121971651 ACN121971651 ACN 121971651ACN-121971651-A

Abstract

The invention discloses a metal-based nanoparticle loaded with traditional Chinese medicines, and a preparation method and application thereof, and relates to the technical field of nano medical materials. The nanoparticle is prepared by taking nickel hydroxide iron zinc aluminum as a multi-metal-based core carrier, carrying out PEG modification and cRGD peptide targeted coupling, and loading curcumin as an active ingredient of a traditional Chinese medicine. The preparation is realized by a multi-step chemical coupling and loading process, and the multi-metal carrier can realize the responsive structural disintegration and the accurate controlled release of curcumin in a weak acidic environment of tumors. The nanoparticle has the functions of active targeting, multi-metal synergistic injury killing and immune regulation, can specifically identify the integrin alpha v beta 3 receptor on the surface of the triple negative breast cancer cells, inhibit proliferation of tumor cells, induce apoptosis, regulate and control immune-related signal paths such as BCR, IL-17 and the like, promote infiltration of immune cells and remodel tumor immune microenvironment. The metal-based nano particles loaded with the traditional Chinese medicine have simple preparation process and high production efficiency, and are suitable for the accurate treatment of triple negative breast cancer.

Inventors

JIAO JUNNA
Lv Zhuangwei
GUO LIWEI
WU HONGYAN
HUANG YANMEI
ZHANG LIN
WANG SHUFANG

Assignees

河南医药大学

Dates

Publication Date: 20260505
Application Date: 20260227

Claims (10)

1. The metal-based nanoparticle loaded with the traditional Chinese medicine is characterized by comprising a multi-metal-based composite carrier, a polyethylene glycol chain segment connected to the multi-metal-based composite carrier, a targeting ligand cRGD peptide coupled to the polyethylene glycol chain segment, and a traditional Chinese medicine active ingredient curcumin loaded in the multi-metal-based composite carrier; wherein the multi-metal matrix composite carrier is nickel hydroxide iron zinc aluminum core-shell nano particles formed by the reaction of nickel salt, ferric salt, zinc salt and aluminum salt in a composite solvent.
2. The traditional Chinese medicine-loaded metal-based nanoparticle according to claim 1, wherein the molar ratio of nickel salt, iron salt, zinc salt and aluminum salt is 1-3:2-4:1-2:0.5-1.5; the nickel salt is nickel chloride, the ferric salt is ferric chloride, the zinc salt is zinc chloride, and the aluminum salt is aluminum chloride.
3. The traditional Chinese medicine-loaded metal-based nanoparticle according to claim 1, wherein the loading concentration of curcumin serving as an active ingredient of the traditional Chinese medicine is 50-150 mug/mg, the encapsulation rate is more than or equal to 85%, the modification amount of cRGD peptide is 10-30 mug/mg nano-carrier, the molecular weight of PEG is 2000-5000 Da, and the modification amount is 50-100 mug/mg nano-carrier.
4. A method for preparing the metal-based nanoparticle loaded with a traditional Chinese medicine according to any one of claims 1 to 3, comprising the steps of: under the condition of low-temperature stirring, the metal salt solution and an alkaline solution are subjected to coprecipitation reaction, and the nickel hydroxide iron zinc aluminum core-shell nano particles are obtained after separation, washing and drying; Step 2, dispersing the core-shell nano particles obtained in the step 1 in an organic solvent, adding an aminosilane coupling agent for reflux reaction, and separating and washing to obtain amination modified nano particles; Step 3, dissolving carboxyl-polyethylene glycol-amino molecules in a buffer solution for activation, then adding the amination nano particles obtained in the step 2 for coupling reaction, and obtaining PEG modified nano particles after separation and washing; step 4, reacting the PEG modified nano particles obtained in the step 3 with a bifunctional crosslinking agent, coupling with a targeting ligand cRGD peptide, and separating and washing to obtain the targeting nano carrier; And 5, dissolving curcumin in a solvent, mixing with the targeting nano-carrier obtained in the step 4, stirring and loading under a light-shielding condition, and finally purifying to obtain the curcumin-loaded metal-based nano-particles.
5. The method for preparing the metal-based nano particles loaded with the traditional Chinese medicine according to claim 4, wherein in the step 1, the alkaline solution is a mixed aqueous solution of sodium citrate and sodium hydroxide, the coprecipitation reaction is carried out in a low-temperature water bath at 4 ℃, the metal salt solution and the alkaline solution are mixed in a slow dropwise manner, after the reaction, stirring is carried out at room temperature for 0.5-1 h, ultrasonic dispersion is carried out for 25-35 min, stirring is carried out at a constant temperature for 25-35 min at 60 ℃, and finally centrifugation is carried out for 10-15 min at a rotating speed of 8000-12000 rpm.
6. The method for preparing the metal-based nanoparticle loaded with the traditional Chinese medicine according to claim 4, wherein in the step 2, the aminosilane coupling agent is APTES, the addition amount of the aminosilane coupling agent is 1% (v/v) of the volume of the dispersion liquid of the core-shell nanoparticle, the reflux reaction is carried out for 5-7 hours at 65-75 ℃, the centrifugal speed is 11000-12000 rpm, and the centrifugal time is 10-15 min.
7. The method for preparing the metal-based nanoparticle loaded with traditional Chinese medicine according to claim 4, wherein in the step 3, the molecular weight of the carboxyl-polyethylene glycol-amino molecule is 2000-5000 Da, EDC and NHS are used in the activation process, the molar ratio of EDC to the carboxyl-polyethylene glycol-amino molecule is 2-5:1, the molar ratio of NHS to EDC is 1:1, and the activation reaction is performed in a buffer solution with pH of 5.0-6.0; The coupling reaction is carried out for 2-4 hours at room temperature; after the reaction is finished, centrifugal separation is carried out, the centrifugal speed is 11000-12000 rpm, and the centrifugal time is 10-20 min.
8. The method for preparing the metal-based nanoparticle loaded with the traditional Chinese medicine according to claim 4, wherein in the step 4, the bifunctional crosslinking agent is SMCC, the adding amount of the bifunctional crosslinking agent is 50-80 mg, the bifunctional crosslinking agent reacts with the PEG-modified nanoparticle for 1-2 hours at room temperature and in a dark condition, then cRGDfk-SH solution with the concentration of 1 mg/mL is added, the reaction is carried out for 1.5-2 hours at the condition of 25 ℃ and the stirring speed of 300-500 rpm, and after the reaction is completed, centrifugal separation is carried out, the centrifugal speed is 11000-12000 rpm, and the centrifugal time is 10-20 min.
9. The method for preparing the metal-based nanoparticle loaded with the traditional Chinese medicine according to claim 4, wherein in the step 5, the curcumin is firstly dissolved in dimethyl sulfoxide, then diluted by PBS buffer solution to obtain curcumin solution with the final concentration of 1 mg/mL, the targeting nano-carrier is mixed with the curcumin solution, and the volume content of the dimethyl sulfoxide in the obtained mixed solution is less than 1%; The mass ratio of the targeting nano-carrier to the curcumin is 1:5; Mixing the targeting nano-carrier with the curcumin solution, and stirring for 8-12 hours at 20-30 ℃ under a light-shielding condition to carry out a load reaction; After the reaction is finished, a dialysis bag with the molecular weight cut-off of 3.5 kDa is adopted for dialysis for 22-24 hours by using agains PBS buffer solution for purification, and the buffer solution is replaced in stages in the dialysis process.
10. Use of the traditional Chinese medicine-loaded metal-based nanoparticle according to any one of claims 1 to 3 for preparing a triple negative breast cancer therapeutic drug.

Description

Metal-based nanoparticle loaded with traditional Chinese medicine and preparation method and application thereof Technical Field The invention belongs to the technical field of nano medicine materials, and particularly relates to a metal-based nanoparticle loaded with traditional Chinese medicines, and a preparation method and application thereof, which are particularly suitable for accurate treatment of triple negative breast cancer and tumor immunity microenvironment regulation. Background Triple Negative Breast Cancer (TNBC) is a very high malignancy subtype in breast cancer, accounting for about 15% -20% of the total breast cancer, and due to lack of definite therapeutic targets such as Estrogen Receptor (ER), progestogen Receptor (PR) and human epidermal growth factor receptor 2 (HER 2), it cannot benefit from endocrine therapy or anti-HER 2 targeted therapy, and clinical therapy mainly depends on chemotherapy. However, the traditional chemotherapy drugs have the problems of poor targeting, strong toxic and side effects, low bioavailability, easy drug resistance and the like, and seriously influence the treatment effect and the survival rate of patients. In recent years, nanotechnology has shown great potential in the field of tumor targeted therapy, and has become an important direction for breaking through the bottleneck of TNBC therapy. The metal-based nanoparticles are widely applied to the construction of drug nanoparticles by virtue of the unique physicochemical properties, good biocompatibility and multifunctional modification potential. However, existing metal-based nano-delivery systems still face a number of technical bottlenecks: (1) The targeting accuracy is insufficient, most nano-carriers only depend on the enhanced permeation and retention Effect (EPR) of passive targeting, and the active targeting recognition capability is lacking, so that the medicine enrichment efficiency at the tumor part is low, and the normal tissue damage is serious; (2) The active ingredients of the traditional Chinese medicine are difficult to stably load and slowly release, and the active ingredients of the traditional Chinese medicine such as curcumin have multi-target anti-tumor and immunoregulation potential, but have poor water solubility, fast in-vivo metabolism and low bioavailability, are easy to aggregate and degrade in nano-carriers, and influence the exertion of the drug effect; (3) The existing system focuses on single drug delivery or local killing, lacks systematic regulation capability on tumor immune microenvironment (TME), and is difficult to break pathological circulation of TNBC immunosuppressive microenvironment; (4) The synergistic effect of metal ions is not fully exerted, the single metal-based nano material has limited functions in aspects of tumor microenvironment response, immune regulation and the like, and the proportion optimization and the synergistic mechanism of the multi-metal composite system are not clear. Meanwhile, TNBC tumor microenvironments generally show immunosuppressive characteristics, and insufficient infiltration of effector T/B cells and abnormal immune-related signal paths lead to extremely low response rate of immunotherapy. Therefore, the development of metal-based nano particles with the functions of active targeting delivery, high-efficiency load of active ingredients of traditional Chinese medicines, multi-metal synergy and tumor immunity micro-environment remodeling becomes a key direction for solving the problem of accurate TNBC treatment. Disclosure of Invention In order to overcome the technical problems that the existing metal-based nano-particles are insufficient in targeting property, low in loading efficiency of active ingredients of traditional Chinese medicines, single in function and incapable of effectively regulating and controlling tumor immunity microenvironment, the invention provides the metal-based nano-particles loaded with traditional Chinese medicines, and a preparation method and application thereof. In order to achieve the above purpose, the invention is realized by adopting the following technical scheme: In a first aspect, the invention provides a metal-based nanoparticle loaded with traditional Chinese medicine, which comprises a multi-metal-based composite carrier, a polyethylene glycol chain segment connected to the multi-metal-based composite carrier, a targeting ligand cRGD peptide coupled to the polyethylene glycol chain segment, and a traditional Chinese medicine active ingredient curcumin loaded in the multi-metal-based composite carrier; wherein the multi-metal matrix composite carrier is nickel hydroxide iron zinc aluminum core-shell nano particles formed by the reaction of nickel salt, ferric salt, zinc salt and aluminum salt in a composite solvent. The invention is further improved in that the molar ratio of the nickel salt to the iron salt to the zinc salt to the aluminum salt is 1-3:2-4:1-2:0.5-1.5; the nickel salt is nic