CN110935019A - PH-responsive multifunctional nano medicine carrier and application thereof in skin diseases - Google Patents
PH-responsive multifunctional nano medicine carrier and application thereof in skin diseases Download PDFInfo
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Abstract
The invention discloses a pH-responsive multifunctional nano drug carrier and application thereof in skin diseases, wherein the carrier takes a gold nanoparticle core-shell structure coated with mesoporous silica as a drug carrier, the drug carrier is firstly carboxylated by a TESPA ethanol solution, then a drug aqueous solution is added into a carboxylated drug carrier aqueous solution, loading of a drug is realized by utilizing a porous structure of the mesoporous silica, then EDC, HCl and NHS are added to activate carboxyl, finally a zinc oxide quantum dot pore blocking agent is added, and the drug-loaded nanoparticles are assembled through electrostatic interaction. The multifunctional nano medicine carrier disclosed by the invention not only realizes the comprehensive combination of targeted medicine transfer, photothermal therapy, chemotherapy and immunotherapy and is applied to the treatment of skin tumors and virus-infected skin diseases, but also increases the medicine loading rate to (33.89 +/-1.64)%, improves the clinical curative effect and makes up the defects of the current traditional treatment and the emerging immunotherapy.
Description
Technical Field
The invention belongs to the technical field of biological medicines, and particularly relates to a pH-responsive multifunctional nano medicine carrier and application thereof in skin diseases.
Background
Malignant tumors of the skin, especially malignant melanoma, have extremely high malignancy degree and are easy to metastasize in early stage, and the 5-year survival rate of patients with advanced melanoma with distant metastasis is less than 5%. The traditional operation, radiotherapy and chemotherapy has the disadvantages of large trauma, poor curative effect, difficult positioning and obvious toxic and side effects. Although the existing targeted molecular therapy (such as BRAF and MEK inhibitors) and immune checkpoint inhibitors (such as anti-CTLA-4 and anti-PD-1 monoclonal antibodies) can improve the overall survival rate of melanoma patients, part of patients are easy to resist drugs and relapse, the tumor metastasis and disease progression cannot be effectively controlled, and the treatment-related side effects are obviously increased. In addition, for various benign tumors of the skin and virus infectious diseases, such as condyloma acuminatum, molluscum contagiosum and the like, the existing therapies of external medicines, refrigeration, photodynamic therapy and the like still have the problems of poor clinical curative effect, easy relapse, strong infectivity, even canceration and the like. Therefore, the search for safe and effective combination therapy means and the realization of combination therapy are key problems to be solved urgently at present.
With the development of nano biotechnology, novel nano drugs using nano materials as carriers have been increasingly used for the prevention, diagnosis and treatment of clinical diseases. Compared with the conventional medicine, the nano medicine carrier has small grain diameter and large specific surface area, can improve the medicine-loading rate and increase the solubility and the stability of the medicine. After surface modification, the nano-drug carrier can also realize drug targeted delivery, improve drug effect and reduce toxic and side effects, and especially the multifunctional integrated nano-system can realize multi-means combined treatment.
Mesoporous silica coated gold nanoparticles (Au @ mSiO)2) The mesoporous silica is a mature drug carrier at present, and the mesoporous silica shell has the advantages of large specific surface area, easy modification, good biocompatibility and the like. The mesoporous silica shell layer is loaded with anti-tumor chemotherapeutic drugs or antiviral drugs, so that the dual combination of photothermal therapy and chemotherapy or antiviral can be realized, and the clinical curative effect is improved. However, Au @ mSiO was used2The problems of low drug loading, prodrug release, poor curative effect and the like exist when the drug is loaded. In addition, the zinc oxide quantum dots (ZnO QDs) have pH response characteristics under weak acidic conditions, the zinc oxide quantum dots (ZnOQDs) are easily decomposed in an aqueous solution at the pH of 5, and meanwhile, the zinc oxide quantum dots (ZnOQDs) have an enrichment effect on Doxorubicin (DOX), so that the zinc oxide quantum dots can be used as a drug carrier, can be combined with the Doxorubicin (DOX), can load the Doxorubicin (DOX) into cells, and can be used for treating cancers. However, the Doxorubicin (DOX) loading capacity of the zinc oxide quantum dots (ZnO QDs) is only 20%, and the Doxorubicin (DOX) release rates under different pH values are different, so that the treatment effect is greatly influenced.
Disclosure of Invention
The invention aims to: aiming at the defects and the defects of the existing nano-drug carrier, the carrier takes gold nanoparticles coated by mesoporous silica as the drug carrier and zinc oxide quantum dots as a pore blocking agent, so that the drug loading capacity is increased while the comprehensive combination of targeted drug delivery, photothermal therapy, chemotherapy and immunotherapy is realized, and the effect of improving the clinical curative effect is achieved.
In order to achieve the purpose, the invention adopts the following technical scheme:
a pH-responsive multifunctional nano drug carrier takes a gold nanoparticle core-shell structure coated by mesoporous silica as a drug carrier, and is characterized in that: the drug carrier is firstly carboxylated by TESPA ethanol solution, then the aqueous solution of the drug is added into the aqueous solution of the carboxylated drug carrier, the loading of the drug is realized by utilizing the porous structure of mesoporous silica, then EDC & HCl and NHS are added to activate the carboxyl, finally the zinc oxide quantum dot pore-plugging agent is added, and the drug-loaded nanoparticles are assembled through electrostatic interaction.
A pH-responsive multifunctional nano-drug carrier, characterized in that: the drug carrier is aminated by an APTES ethanol solution, then the drug aqueous solution is added into the aminated drug carrier aqueous solution, the loading of the drug is realized by utilizing the porous structure of mesoporous silica, finally, the copper sulfide nano particle pore plugging agent is added, and the drug-loaded nano particles are assembled through electrostatic interaction.
Further, the gold nanoparticles include, but are not limited to, gold nanospheres, gold nanorods, gold nanocages, and gold nanocoards, preferably gold nanospheres.
Furthermore, in the multifunctional nano drug carrier taking copper sulfide nano particles (CuxSNPs) as the pore blocking agent, in order to improve the stability of the nano assembly, PEG-SH can be added to obtain the PEG modified assembly.
Further, the load drug can be an anti-tumor chemotherapeutic drug or an antiviral agent.
Furthermore, the anti-tumor chemotherapeutic drugs include adriamycin, paclitaxel, podophyllotoxin, dacarbazine and the like, and preferably adriamycin.
Further, the antiviral agent comprises 5-FU, interferon, Polyphenon E, ammonium trichloro-hoof acid (AS101), GS-9191, etc., preferably interferon.
Furthermore, the adriamycin-loaded multifunctional medicine carrier can effectively inhibit the growth of tumors and lung metastasis.
Furthermore, the interferon-loaded multifunctional drug carrier can realize the combination of drug interference, photothermal therapy and immunotherapy on skin virus infectious diseases.
Further, the application method of the pH-responsive multifunctional nano-drug carrier is that ① skin lesions are injected with the pH-responsive multifunctional nano-drug carrier, and ② excitation light with specific wavelength is given to the skin lesion parts after a certain time.
Compared with the prior art, the invention has the beneficial effects that:
1) the multifunctional nano medicine carrier takes the gold nano particles coated by the mesoporous silica as the medicine carrier, and also takes the zinc oxide quantum dots as the pore blocking agent, so that the comprehensive combination of targeted medicine transfer, photo-thermal treatment, chemotherapy and immunotherapy is realized, the multifunctional nano medicine carrier is applied to the treatment of skin tumors and virus-infected skin diseases, the medicine loading rate is increased to (33.89 +/-1.64)%, the clinical curative effect is improved, and the defects of the current traditional treatment and the emerging immunotherapy are overcome;
2) the multifunctional nano medicine carrier has low production cost, simple and convenient synthesis process, high medicine carrying rate and easy popularization and application;
3) in the multifunctional nano medicine carrier, the photothermal effect of the gold nanoparticles and the medicine carried by the gold nanoparticles can realize the combined treatment of photothermal treatment and chemotherapy; the pH response characteristic of the zinc oxide quantum dots can realize the targeted delivery of the drug, and finally, the nano drug carrier can effectively inhibit the growth and metastasis of tumors;
4) the multifunctional nano medicine carrier can be applied to treatment of various diseases, and specifically comprises ① skin benign tumors and precancerous lesions, such as seborrheic keratosis, bowenoid papulosis, solar keratosis, infantile hemangioma, vascular malformation and the like, ② skin malignant tumors, such as malignant melanoma, skin lymphoma, skin squamous cell carcinoma, basal cell carcinoma and the like, ③ virus-infected skin diseases, such as condyloma acuminatum, verruca vulgaris, verruca plana infectans and the like, and is particularly suitable for malignant melanoma and condyloma acuminatum.
Drawings
Fig. 1 is a schematic diagram (a) and an action mechanism diagram (b) of a multifunctional nano drug carrier loaded with adriamycin.
FIG. 2 is the characterization result of the multifunctional nano-drug carrier loaded with adriamycin, wherein, (a) TEM image of ZnO QDs; (b-c) AuNP @ mSiO2TEM and SEM images of (a); (d) AuNP @ mSiO2TEM image of ZnO; (e-h) AuNP @ mSiO2STEM-EDX diagram of Zn element, Si element, O element and Au element in ZnO.
Fig. 3 is a graph showing the results of the doxorubicin-loaded multifunctional nano-drug carrier for in vivo inhibition of tumor growth in melanoma model mice, specifically the results of (a) tumor weight and (b) tumor volume changes in melanoma model mice after treatment.
FIG. 4 is a graph showing the results of doxorubicin-loaded multifunctional nano-drug carrier inducing tumor necrosis and inhibiting lung metastasis, specifically HE staining and Ki-67 staining of in situ tumors and lung tissues of tumor-bearing mice after treatment.
Detailed Description
The technical solutions of the present invention are further described in detail below with reference to specific examples to facilitate understanding for those skilled in the art. It should be noted that the following examples are only for explaining and illustrating the present invention and are not to be construed as limiting the scope of the present invention.
Example 1
The multifunctional nanometer medicine carrier loaded with adriamycin has the following specific preparation method:
1) preparation of mesoporous silica-coated gold nanospheres (AuNP @ mSiO) by one-pot method2) A drug carrier of "core-shell" structure;
2) preparing water-soluble zinc oxide quantum dots (ZnO QDs or ZnO-NH) by adopting a sol-gel method2)；
3) Adding a TESPA ethanol solution into the drug carrier prepared in the step 1) to prepare carboxylated AuNP @ mSiO2；
4) To AuNP @ mSiO prepared in step 3)2Adding adriamycin (DOX) aqueous solution into the aqueous solution of-COOH, stirring for 24 hours in a dark place, and realizing the load of the adriamycin (DOX) by utilizing the porous structure of the mesoporous silica;
5) to the DOX-loaded AuNP @ mSiO prepared in step 4)2EDC, HCl and NHS are added into-COOH, after carboxyl is activated, ZnOQDs prepared in the step 2) are added, and the drug-loaded nano particles are assembled through electrostatic interaction, so that the pH-responsive nano drug-loaded body (AuNP @ mSiO) with gold nanospheres as the core is obtained2@ DOX-ZnO), as shown in FIG. 1- (a).
The prepared AuNP @ mSiO is subjected to electron microscopy, ultraviolet visible light-spectrophotometer, Fourier infrared spectrometer and the like2@ DOX-ZnO, as shown in FIG. 2.
Example 2
AuNP@mSiO2Application of @ DOX-ZnO in inhibiting growth of melanoma and lung metastasis
1) The in vivo experimental groups were as follows: PBS, PBS + NIR, DOX, AuNP @ mSiO2-ZnO、AuNP@mSiO2@DOX-ZnO、AuNP@mSiO2-ZnO+NIR、AuNP@mSiO2The @ DOX-ZnO + NIR and illumination group adopts 655nm laser irradiation (1.0W/cm)240 s). The treatment is performed once every 3 days, and the treatment is performed 3 times in total. Recording the length and width of the tumor by a vernier caliper every other day, and calculating the volume of the tumor;
2) the experiment was terminated from first treatment to day 16 post-treatment, tumor weights were measured and mouse tumors and lung tissue were retained for immunohistochemical staining for HE and Ki-67.
The experimental results show that: AuNP @ mSiO despite lower irradiation power and treatment time2@ DOX-ZnO was effective in inhibiting tumor growth (as shown in FIG. 3) and lung metastasis (as shown in FIG. 4).
Example 3
The multifunctional nano-carrier loaded with interferon is prepared by the following specific steps:
1) preparation of mesoporous silica-coated gold nanospheres (AuNP @ mSiO) by one-pot method2) A drug carrier of "core-shell" structure;
2) preparing water-soluble zinc oxide quantum dots (ZnO QDs or ZnO-NH) by adopting a sol-gel method2)；
3) Adding a TESPA ethanol solution into the drug carrier prepared in the step 1) to prepare carboxylated AuNP @ mSiO2；
4) To AuNP @ mSiO prepared in step 3)2Adding an Interferon (IFN) -gamma aqueous solution into the aqueous solution of the-COOH, stirring for 24 hours in a dark place, and realizing the loading of the IFN by utilizing the porous structure of the mesoporous silica;
5) to the IFN-loaded AuNP @ mSiO prepared in step 4)2EDC & HCl and NHS are added into-COOH, after carboxyl is activated, ZnOQDs prepared in the step 2) are added, and the drug-loaded nano particles are assembled through electrostatic interaction, so that the pH-responsive nano drug-loaded body (AuNP @ mSiO) with gold nano particles as the core is obtained2@IFNγ-ZnO)。
Example 4
The adriamycin-loaded multifunctional nano medicine carrier taking the gold nanorods as the core is prepared by the following specific steps:
1) preparing gold nanorods by adopting a seed growth method, adding a sodium hydroxide solution and a TEOS methanol solution into the prepared gold nanorods, and reacting to obtain a mesoporous silica coated gold nanorod drug carrier (AuNR @ mSiO) with a core-shell structure2)；
2) Preparing water-soluble zinc oxide quantum dots (ZnO QDs or ZnO-NH) by adopting a sol-gel method2)；
3) Adding a TESPA ethanol solution into the drug carrier prepared in the step 1) to prepare carboxylated AuNP @ mSiO2；
4) To AuNP @ mSiO prepared in step 3)2Adding adriamycin (DOX) aqueous solution into the aqueous solution of-COOH, stirring for 24 hours in a dark place, and realizing the load of the adriamycin (DOX) by utilizing the porous structure of the mesoporous silica;
5) to the DOX-loaded AuNP @ mSiO prepared in step 4)2EDC & HCl and NHS are added into-COOH, after carboxyl is activated, ZnOQDs prepared in the step 2) are added, and the drug-loaded nanoparticles are assembled through electrostatic interaction, so that the pH-responsive nano drug-loaded body (AuNR @ mSiO2@ DOX-ZnO) with gold nanorods as the core is obtained.
2) The nano drug-loading system (AuNR @ mSiO2@ DOX-ZnO) with the gold nanorods as the core is injected into the skin lesion, and the skin lesion is irradiated after a certain time point, so that the combination of drug interference, photothermal therapy and immunotherapy to the skin tumor is realized.
Example 5
The multifunctional nano medicine carrier with copper sulfide nano particles (CuxSNPs) as a pore blocking agent has the specific preparation method that:
1) preparation of mesoporous silica-coated gold nanospheres (AuNP @ mSiO) by one-pot method2) A drug carrier of "core-shell" structure;
2) adding an APTES ethanol solution into the drug carrier prepared in the step 1) to prepare aminated AuNP @ mSiO2；
3) To AuNP @ mSiO prepared in step 2)2-NH2Adding adriamycin (DOX) aqueous solution into the aqueous solution, stirring for 24 hours in the dark, realizing the load of the adriamycin (DOX) by utilizing the porous structure of the mesoporous silica, and assembling the drug-loaded nano particles through electrostatic action to obtain the nano drug-loaded system (AuNP @ mSiO) taking the gold nanospheres as the core2@ DOX-CuxS), in order to improve the stability of the nano-assembly, PEG-SH can be added to obtain PEG modified assembly (AuNP @ mSiO)2@DOX-CuxS-PEG)。
Example 6
AuNP@mSiO2@IFNγ-ZnO、AuNR@mSiO2@ DOX-ZnO and AuNP @ mSiO2The application of @ DOX-CuxS-PEG in virus-infected skin diseases: mixing AuNP @ mSiO2@IFNγ-ZnO、AuNR@mSiO2@ DOX-ZnO or AuNP @ mSiO2The @ DOX-CuxS-PEG is injected into the skin lesion, and the skin lesion is irradiated after 3 to 6 hours, so that the combination of drug interference, photothermal therapy and immunotherapy to the skin virus infectious diseases is realized.
Claims (10)
1. A pH-responsive multifunctional nano drug carrier takes a gold nanoparticle core-shell structure coated by mesoporous silica as a drug carrier, and is characterized in that: the drug carrier is firstly carboxylated by TESPA ethanol solution, then the aqueous solution of the drug is added into the aqueous solution of the carboxylated drug carrier, the loading of the drug is realized by utilizing the porous structure of mesoporous silica, then EDC & HCl and NHS are added to activate the carboxyl, finally the zinc oxide quantum dot pore-plugging agent is added, and the drug-loaded nanoparticles are assembled through electrostatic interaction.
2. A pH-responsive multifunctional nano drug carrier takes a gold nanoparticle core-shell structure coated by mesoporous silica as a drug carrier, and is characterized in that: the drug carrier is aminated by an APTES ethanol solution, then the drug aqueous solution is added into the aminated drug carrier aqueous solution, the loading of the drug is realized by utilizing the porous structure of mesoporous silica, finally, the copper sulfide nano particle pore plugging agent is added, and the drug-loaded nano particles are assembled through electrostatic action.
3. The pH-responsive multifunctional nano-drug carrier according to claim 1 or 2, wherein: the gold nanoparticles comprise any one of gold nanospheres, gold nanorods, gold nanocages or gold nanocoards.
4. The pH-responsive multifunctional nano-drug carrier according to claim 3, wherein: the gold nanoparticles are gold nanospheres.
5. The pH-responsive multifunctional nano-drug carrier according to claim 1, wherein: the load drug is an anti-tumor chemotherapeutic drug or an antiviral preparation.
6. The pH-responsive multifunctional nano-drug carrier of claim 1, wherein: and adding PEG-SH into the multifunctional nano medicine carrier to obtain a PEG modified assembly.
7. The pH-responsive multifunctional nano-drug carrier according to claim 5, wherein: the anti-tumor chemotherapeutic drug is adriamycin.
8. The pH-responsive multifunctional nano-drug carrier according to claim 5, wherein: the antiviral agent is interferon.
9. The pH-responsive multifunctional nano-drug carrier according to claim 7, wherein: the multifunctional drug carrier loaded with the adriamycin is applied to inhibiting the growth of melanoma and lung metastasis.
10. The pH-responsive multifunctional nano-drug carrier according to claim 8, wherein: the interferon-loaded multifunctional drug carrier is applied to virus-infected skin diseases.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107412195A (en) * | 2017-05-08 | 2017-12-01 | 华中科技大学 | A kind of antineoplastic drug carrier material of pH responses and its preparation and application |
| CN109893661A (en) * | 2019-04-28 | 2019-06-18 | 南京邮电大学 | Gold nanorods/zinc oxide/mesoporous silicon dioxide nano the composite material and preparation method and application of adriamycin insertion |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107412195A (en) * | 2017-05-08 | 2017-12-01 | 华中科技大学 | A kind of antineoplastic drug carrier material of pH responses and its preparation and application |
| CN109893661A (en) * | 2019-04-28 | 2019-06-18 | 南京邮电大学 | Gold nanorods/zinc oxide/mesoporous silicon dioxide nano the composite material and preparation method and application of adriamycin insertion |
Non-Patent Citations (2)
| Title |
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| 刘丽平: "多功能金纳米粒子载药体系的构建及其抗肿瘤应用", 《中国学位论文全文数据库》 * |
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