Preparation method of transdermal self-assembly nanomedicine and its products and applications

Abstract

The invention discloses a preparation method of a transdermal self-assembly nanomedicine: by the self-assembly site reaction of 6-mercaptopurine and zinc acetate, programmed death receptor 1 is added to the reaction system, and the self-assembly nanomedicine 6-MP / Zn / PD-1 loaded with 6-MP and PD-1 is prepared; Arginine-rich dendrimer (GR) is modified on the surface to prepare a transdermal self-assembly nanomedicine 6-MP / Zn / PD-1 / GR with transdermal ability; 6-MP / Zn / PD-1 / GR can also be incorporated into a gel for transdermal administration. The preparation method provided by the present invention is extremely simple, and the transdermal self-assembly nanomedicine prepared simultaneously coats therapeutic antibodies therein, achieves the synergistic treatment of chemotherapy and immunity, and can achieve transdermal administration, which can be applied to prepare a drug for transdermal administration and can effectively deliver drugs through the skin.

Description

Technical Field

[0001] The present invention belongs to the field of preparation of biomedical materials, in particular to a preparation method of a transdermal self-assembly nano drug and its product and application. Background Art

[0002] Topical drug delivery refers to a therapeutic approach that applies medication directly to the site of disease, using methods such as bandaging, injection, application, spray, or patch, depending on the pathogenesis of the disease. Unlike systemic treatments such as oral or intravenous injections, topical drug delivery offers unique advantages for combating many superficial diseases or tumors, including high drug availability, high therapeutic efficiency, and minimal side effects. Transdermal drug delivery systems are a newly developed new drug delivery system. (See: MR Prausnitz, R. Langer. Nat. Biotechnol. 2008, 26, 1261-1268.) After topical administration, transdermal drug delivery systems penetrate the skin at a constant rate, entering the body's circulation and producing systemic or local therapeutic effects. Advantages include drug absorption that is unaffected by factors such as the pH of the digestive tract and the complex drug circulation process; avoidance of the first-pass effect in the liver; overcoming adverse reactions caused by excessively rapid absorption and resulting high blood drug concentrations; and the ability to continuously control the delivery rate, allowing for flexible dosing. Transdermal drug delivery systems have become an effective treatment for tumors, particularly superficial melanomas. See: P. Kriplani, K. Guarve. Recent Pat. Anticancer Drug Discov. 2022, 17, 253-267. The active ingredients in topical transdermal preparations need to penetrate the skin barrier into the subcutaneous tissue or bloodstream to achieve their therapeutic effects. However, the human skin has a stratum corneum, which is a difficult barrier for most drugs to penetrate. Transdermal absorption requires breaking through the stratum corneum barrier, and the rate and depth of drug penetration will affect the drug's efficacy. Many drugs have difficulty penetrating the skin or the amount of penetration is difficult to reach therapeutic levels. Therefore, promoting transdermal drug penetration through appropriate methods has become a key research topic in transdermal drug delivery systems. See: CY Deng, R. Zhuang, ZY Ying, JS Tu, XH Xu, CM Sun, L. Jiang, Adv. Funct. Mater. 2022, 32, 2206876. Among technologies that promote transdermal drug penetration, non-invasive nanocarrier-based drug delivery systems are a proven approach. Nanoemulsions and liposomes, for example, have been used for transdermal drug delivery. These nanocarriers can help some drugs that are difficult to penetrate transdermally enter the body and exert their efficacy. See: T. Jiang, T. Wang, T. Li, Y. Ma, S. Shen, B. He, R. Mo, ACS Nano 2018, 12, 9693; P. Pandi, A. Jain, N. Kommineni, M. Ionov, M. Bryszewska, W. Khan, Int. J. PHarm. 2018, 550, 240-250. However, promoting drug penetration into subcutaneous tissue or subcutaneous tumors remains a major therapeutic challenge and a key challenge in the development of permeation-enhancing technologies within transdermal delivery.Currently, there are very few non-invasive transdermal treatments that can help drugs cross the stratum corneum. See: X. Zhang, X. Xu, Y. Li, C. Hu, Z. Zhang, Z. Gu, Adv. Mater. 2018, 30, 1707-240. In addition, metal-organic polymers, constructed by self-assembly of metal ions and organic ligands, have been developed for drug delivery. However, these metal-organic polymers do not have therapeutic capabilities and usually achieve treatment by encapsulating other chemotherapy drugs, which makes them somewhat complex. See: YT Qin, H. Peng, WY Li, YK Zhang, ACS Appl. Mater. Interfaces, 2020, 12, 24585-24598.

[0003] Based on this, it is crucial to develop a simple synthetic method for preparing transdermal self-assembled nanomedicines. Summary of the Invention

[0004] The purpose of the present invention is to provide a method for preparing a transdermal self-assembled nanomedicine, its products and applications. The preparation method provided by the present invention is extremely simple. The prepared transdermal self-assembled nanomedicine simultaneously encapsulates therapeutic antibodies therein, thereby achieving synergistic treatment of chemotherapy and immunity, and can realize transdermal administration. It can be used in the preparation of drugs for transdermal administration and can effectively deliver drugs through the skin.

[0005] The present invention provides the following technical solutions:

[0006] A method for preparing a transdermal self-assembly nanomedicine, comprising:

[0007] 1) Dissolve 6-MP in a buffer solution, add ZA and PD-1, and stir at room temperature to react to obtain a self-assembled nanodrug 6-MP / Zn / PD-1;

[0008] 2) 6-MP / Zn / PD-1 was dispersed in a buffer solution, DF-PEG2000-N3 was added, and the mixture was stirred at room temperature to obtain a self-assembled nanodrug 6-MP / Zn / PD-1 / N3 with surface modified azide groups;

[0009] 3) Boc-Arg(Boc)-OPFP was mixed with G5, a catalyst was added, and the mixture was reacted at room temperature and then deprotected to obtain the arginine-rich dendrimer GR;

[0010] 4) GR was dissolved in a buffer solution, and NHS-PEG2000-DBCO was added and stirred at room temperature to obtain an arginine-rich dendrimer GR / DBCO with surface modification of dibenzocyclooctyne;

[0011] 5) Dissolving 6-MP / Zn / PD-1 / N3 in a buffer solution, adding GR / DBCO, and stirring at room temperature to react to obtain a transdermal self-assembled nanodrug 6-MP / Zn / PD-1 / GR;

[0012] Among them, 6-MP is 6-mercaptopurine, ZA is zinc acetate, PD-1 is sequenced death receptor 1, DF-PEG2000-N3 is benzaldehyde-polyethylene glycol 2000-azide, Boc-Arg(Boc)-OPFP is pentafluorophenol-activated tri-tert-butyloxycarbonylarginine, G5 is the fifth-generation symmetrical pentamethine cyanine dye dendrimer, and NHS-PEG2000-DBCO is active ester-polyethylene glycol 2000-dibenzocyclooctyne.

[0013] The preparation method provided by the present invention involves a self-assembly reaction of 6-mercaptopurine (6-MP) and zinc acetate (ZA), and the addition of programmed death receptor 1 (PD-1) to the reaction system to prepare a self-assembled nanodrug (6-MP / Zn / PD-1) loaded with 6-MP and PD-1; an arginine-rich dendrimer (GR) is modified on the surface to prepare a self-assembled nanodrug (6-MP / Zn / PD-1 / GR) with transdermal ability; finally, 6-MP / Zn / PD-1 / GR is incorporated into a hydrogel for transdermal administration. The transdermal self-assembled nanodrug prepared by the present invention has been shown to be capable of transdermal administration using the Franz transdermal diffusion model; its inherent red fluorescence can be used for visual observation; and degradation in a simulated tumor microacid environment can achieve the effective release of 6-MP and PD-1.

[0014] The present invention directly constructs a nanomedicine by coordinating the chemotherapy drug 6-mercaptopurine with zinc acetate, and simultaneously encapsulates the therapeutic antibody therein, thereby achieving synergistic chemotherapy and immunotherapy. This method of directly using the chemotherapy drug as a coordinating molecule is extremely simple.

[0015] Preferably, in step 1), the usage ratio of 6-MP, ZA and PD-1 is: 1 mol: 1.25 mol: 5-10 μmol.

[0016] In step 1), a phosphate buffer solution with a pH of 8.5 was used, magnetic stirring was performed for 8–12 h, centrifuged, and deionized water was added to wash the mixture three times, the mixture was collected by centrifugation, and freeze-dried to obtain 6-MP / Zn / PD-1.

[0017] Preferably, in step 2), the dosage ratio of 6-MP / Zn / PD-1 and DF-PEG2000-N3 is: 20 mg: 1-3 mg.

[0018] In step 2), a phosphate buffer solution with a pH of 7.4 was used, stirred at room temperature for 16–24 h, and then centrifuged. Deionized water was added and washed three times, and the mixture was collected by centrifugation and freeze-dried to obtain 6-MP / Zn / PD-1 / N3.

[0019] Preferably, in step 3), the catalysts diisopropylethylenediamine (DIPEA) and trifluoroacetic acid (TFA) are used for deprotection, and the dosage ratio of Boc-Arg(Boc)-OPFP, G5, DIPEA, and TFA is: 20-80 mg: 2 mg: 3-10 μL: 1-2 mL.

[0020] In step 3), the reaction is carried out at room temperature for 2-4 hours, and the product is purified by precipitation with anhydrous ether to obtain GR.

[0021] Preferably, in step 4), the dosage ratio of GR to NHS-PEG2000-DBCO is: 10 mg: 1-3 mg.

[0022] In step 4), a phosphate buffer solution with a pH of 7.4 is used, stirred at room temperature for 16-24 hours, centrifuged using an ultrafiltration centrifuge tube, repeatedly washed three times with deionized water, and freeze-dried to obtain GR / DBCO.

[0023] Preferably, in step 5), the usage ratio of 6-MP / Zn / PD-1 / N3 to GR / DBCO is: 10 mg: 2-5 mg.

[0024] In step 5), a phosphate buffer solution with a pH of 7.4 was used, stirred at room temperature for 2-6 hours, centrifuged, and washed three times with deionized water. The mixture was collected by centrifugation and freeze-dried to obtain 6-MP / Zn / PD-1 / GR.

[0025] The preparation method further comprises step 6): mixing 6-MP / Zn / PD-1 / GR with fibrinogen and thrombin to obtain a transdermal self-assembled nano drug gel Gel@6-MP / Zn / PD-1 / GR.

[0026] Preferably, in step 6), the dosage ratio of 6-MP / Zn / PD-1 / GR, fibrinogen and thrombin is: 2-10 mg: 1 mL: 1 mL, the thrombin concentration is 2 mg / mL, and the fibrinogen concentration is 10 mg / mL.

[0027] The transdermal self-assembled nano-drug gel Gel@6-MP / Zn / PD-1 / GR provided by the present invention can be degraded in the slightly acidic environment of the tumor to release 6-MP and PD-1, effectively killing melanoma cells.

[0028] The present invention also provides a transdermal self-assembly nanomedicine obtained according to the above preparation method.

[0029] The present invention also provides an application of the above transdermal self-assembly nanomedicine in preparing transdermal drug delivery.

[0030] The present invention also provides a use of the above-mentioned transdermal self-assembled nanomedicine in preparing a transdermal drug for treating tumors. The tumor is melanoma.

[0031] Advantages and beneficial effects of the present invention:

[0032] 1) Arginine-rich dendrimers have excellent red fluorescence properties. When modified on the surface, nanomedicines also exhibit excellent fluorescence properties, which can be used for visual monitoring and observation of transdermal drug delivery.

[0033] 2) The nanomedicine prepared by the present invention has a strong transdermal ability due to the presence of a large number of arginine surface groups; and the Franz transdermal diffusion model has demonstrated that it can achieve transdermal administration;

[0034] 3) 6-mercaptopurine (6-MP) and zinc acetate are self-assembled to form nanomedicines. 6-MP itself has therapeutic effects, eliminating the need for specialized chemotherapeutic drug encapsulation. PD-1 is also encapsulated within the nanomedicine, achieving synergistic chemotherapy and immunotherapy.

[0035] 4) The transdermal self-assembled nanomedicine prepared by the present invention is embedded in the gel and can be administered by coating on the skin surface, which is flexible and convenient. BRIEF DESCRIPTION OF THE DRAWINGS

[0036] Figure 1 (a) Transmission electron microscopy image and (b) Zeta potential image of the transdermal self-assembled nanodrug 6-MP / Zn / PD-1 / GR (6-MP / Zn / PD-1 was the negative control group).

[0037] Figure 2 This is a scanning electron microscopy image of the transdermal self-assembled nanodrug gel Gel@6-MP / Zn / PD-1 / GR.

[0038] Figure 3 This is the infrared spectrum of the transdermal self-assembled nanodrug 6-MP / Zn / PD-1 / GR.

[0039] Figure 4 Schematic diagram of the Franz transdermal diffusion model.

[0040] Figure 5 Cumulative release curves of (a) 6-MP and (b) PD-1 of Gel@6-MP / Zn / PD-1 / GR in the Franz transdermal diffusion model (Gel@6-MP and Gel@PD-1 were negative control groups).

[0041] Figure 6This is a bar graph showing the toxicity test of different Gel@6-MP / Zn / PD-1 / GR formulations on melanoma cells B16F10 (Gel, Gel@6-MP and Gel@PD-1 were negative control groups). DETAILED DESCRIPTION

[0042] Example 1:

[0043] The preparation method of the transdermal self-assembling nanomedicine provided in this embodiment specifically comprises the following steps:

[0044] 1) Preparation of self-assembled nanomedicine (6-MP / Zn / PD-1): 1 mol of 6-MP was dissolved in 10 mL of phosphate buffer solution (pH = 8.5), 1.25 mol of zinc acetate (ZA) and 5 μmol of programmed death receptor 1 (PD-1) were added, and the mixture was incubated at room temperature with magnetic stirring for 8–12 h, centrifuged, and repeatedly washed with deionized water three times. The mixture was collected by centrifugation and lyophilized to obtain 6-MP / Zn / PD-1.

[0045] 2) Preparation of self-assembled nanodrugs modified with azide groups (6-MP / Zn / PD-1 / N3): 20 mg of 6-MP / Zn / PD-1 was dispersed in phosphate buffer solution at pH 7.4, 1 mg of benzaldehyde-polyethylene glycol 2000-azide (DF-PEG2000-N3) was added, and the mixture was stirred at room temperature for 16–24 h. The mixture was centrifuged and washed three times with deionized water. The mixture was collected by centrifugation and lyophilized to obtain 6-MP / Zn / PD-1 / N3.

[0046] 3) Preparation of arginine-rich dendrimer (GR): 20 mg of tri-tert-butyloxycarbonylarginine (Boc-Arg(Boc)-OPFP) activated with pentafluorophenol was mixed with 2 mg of the fifth-generation symmetrical pentamethine cyanine dye dendrimer G5. 3 μL of the catalyst diisopropylethylenediamine (DIPEA) was added and the mixture was reacted at room temperature for 2-4 h. The mixture was then deprotected with 1 mL of trifluoroacetic acid (TFA) and purified by precipitation with anhydrous ether to obtain GR.

[0047] 4) Preparation of surface-modified dibenzocyclooctyne-rich arginine dendrimers (GR / DBCO): 10 mg of GR was dissolved in a phosphate buffer solution at pH 7.4, 1 mg of the active ester polyethylene glycol 2000-dibenzocyclooctyne (NHS-PEG2000-DBCO) was added, and the mixture was stirred at room temperature for 16-24 hours. The mixture was then centrifuged using an ultrafiltration centrifuge tube, washed three times with deionized water, and lyophilized to obtain GR / DBCO.

[0048] 5) Preparation of a transdermal nanodrug (6-MP / Zn / PD-1 / GR): 10 mg of 6-MP / Zn / PD-1 / N3 was dissolved in a phosphate buffer solution (pH 7.4), 2 mg of GR / DBCO was added, and the mixture was stirred at room temperature for 2-6 hours, centrifuged, and repeatedly washed three times with deionized water. The mixture was collected by centrifugation and lyophilized to obtain 6-MP / Zn / PD-1 / GR.

[0049] 6) Preparation of transdermal self-assembled nanodrug gel preparation (Gel@6-MP / Zn / PD-1 / GR). 2 mg of 6-MP / Zn / PD-1 / GR nanodrug was mixed with fibrinogen (1 mL, 10 mg / mL) and thrombin (1 mL, 2 mg / mL) to obtain Gel@6-MP / Zn / PD-1 / GR.

[0050] Figure 1 (a) Transmission electron microscopy image and (b) Zeta potential image of the transdermal self-assembled nanodrug 6-MP / Zn / PD-1 / GR prepared in this example (6-MP / Zn / PD-1 is a negative control group). Figure 1 It shows that the particle size of 6-MP / Zn / PD-1 / GR nanodrug is about 47nm and the Zeta potential is about +40.2mV.

[0051] Figure 2 is a scanning electron microscope image of the transdermal self-assembled nano-drug gel Gel@6-MP / Zn / PD-1 / GR prepared in this example, Figure 2 Shown in the middle: The gel has a porous network structure.

[0052] Figure 3 is the infrared spectrum of the transdermal self-assembled nanomedicine 6-MP / Zn / PD-1 / GR prepared in this example, Figure 3 Display: at 3100cm -1 , 1551cm -1 , 1279cm -1 , 3429cm -1 , 1345cm -1 , 1227cm -1 , 1124cm -1 , 1011cm -1 , 939cm -1 , 872cm -1 , 499cm -1 The characteristic absorption peaks of PD-1, GR and 6-MP appeared respectively.

[0053] The Gel@6-MP / Zn / PD-1 / GR prepared in this example was subjected to a transdermal diffusion cell test using the following method: Figure 4The Franz transdermal diffusion model shown is as follows: the mouse skin is clamped at the skin placement part of the device, Gel@6-MP / Zn / PD-1 / GR, Gel@6-MP and Gel@PD-1 are respectively incorporated into the hydrogels prepared with thrombin and fibrinogen, and applied to the administration site, a pH = 6.0 phosphate buffer is added to the receiving cell, magnetic stirring is performed, and samples are taken with a sampling needle at certain time intervals to test the concentrations of 6-MP and PD-1.

[0054] Figure 5 It is the Gel@6-MP of Gel@6-MP / Zn / PD-1 / GR in Franz transdermal diffusion model ( Figure 5 Middle a) and Gel@PD-1( Figure 5 b) Cumulative release curve (Gel@6-MP and Gel@PD-1 are negative control groups), Figure 5 The results show that Gel@6-MP / Zn / PD-1 / GR has the strongest transdermal ability and can penetrate the skin into the receptor pool more than Gel@6-MP and Gel@PD-1, thereby releasing more 6-MP and PD-1 into the receptor pool.

[0055] Among them, the preparation method of Gel@6-MP and Gel@PD-1 in the negative control group is: mixing 6-MP with fibrinogen and thrombin, and mixing PD-1 with fibrinogen and thrombin. The mixing method is the same as the method of incorporating 6-MP / Zn / PD-1 / GR into the gel.

[0056] The toxicity of Gel@6-MP / Zn / PD-1 / GR prepared in this example on melanoma cells B16F10 was tested. Specifically, Gel@6-MP / Zn / PD-1 / GR containing different concentrations of 6-MP / Zn / PD-1 / GR were applied to the administration site of the Franz transdermal diffusion model (Gel, Gel@6-MP and Gel@PD-1 were negative control groups). 24 hours after transdermal administration, the drug in the receiving pool was added to a 96-well plate cultured with melanoma cells B16F10 and incubated with the cells. After a certain period of time, the cell survival rate was measured.

[0057] Figure 6 This is a bar graph showing the toxicity of different Gel@6-MP / Zn / PD-1 / GR concentrations on melanoma B16F10 cells (Gel, Gel@6-MP, and Gel@PD-1 serve as negative control groups). The graph shows that Gel@6-MP / Zn / PD-1 / GR has the strongest cytotoxicity.

[0058] Example 2:

[0059] A method for preparing a transdermal self-assembly nanomedicine, wherein the synthesis steps are substantially the same as those in Example 1, except that: in step 1), 8 μmol of programmed death receptor 1 (PD-1) is added.

[0060] The characterization and application results of the final prepared transdermal self-assembled nanomedicine are similar to those in Example 1.

[0061] Example 3:

[0062] A method for preparing a transdermal self-assembly nanomedicine, the synthesis steps are basically the same as those in Example 1, except that: in step 1), 10 μmol of programmed death receptor 1 (PD-1) is added.

[0063] The characterization and application results of the final prepared transdermal self-assembled nanomedicine are similar to those in Example 1.

[0064] Example 4:

[0065] A method for preparing a transdermal self-assembly nanomedicine, the synthesis steps are basically the same as those in Example 1, except that: in step 2), 2 mg of benzaldehyde-polyethylene glycol 2000-azide (DF-PEG2000-N3) is added.

[0066] The characterization and application results of the final prepared transdermal self-assembled nanomedicine are similar to those in Example 1.

[0067] Example 5:

[0068] A method for preparing a transdermal self-assembly nanomedicine, the synthesis steps are basically the same as those in Example 1, except that: in step 2), 3 mg of benzaldehyde-polyethylene glycol 2000-azide (DF-PEG2000-N3) is added.

[0069] The final preparation of transdermal self-assembled nanomedicine and its application results are similar to those in Example 1.

[0070] Example 6:

[0071] A method for preparing a transdermal self-assembly nanomedicine, wherein the synthesis steps are substantially the same as those in Example 1, except that: in step 3) 40 mg of tri-tert-butyloxycarbonyl arginine (Boc-Arg(Boc)-OPFP) activated with pentafluorophenol is mixed with 2 mg of a fifth-generation symmetrical pentamethine cyanine dye dendrimer G5.

[0072] The characterization and application results of the final prepared transdermal self-assembled nanomedicine are similar to those in Example 1.

[0073] Example 7:

[0074] A method for preparing a transdermal self-assembly nanomedicine, wherein the synthesis steps are substantially the same as those in Example 1, except that: in step 3) 60 mg of tri-tert-butyloxycarbonyl arginine (Boc-Arg(Boc)-OPFP) activated with pentafluorophenol is mixed with 2 mg of a fifth-generation symmetrical pentamethine cyanine dye dendrimer G5.

[0075] The characterization and application results of the final prepared transdermal self-assembled nanomedicine are similar to those in Example 1.

[0076] Example 8:

[0077] A method for preparing a transdermal self-assembly nanomedicine, wherein the synthesis steps are substantially the same as those in Example 1, except that: in step 3) 80 mg of tri-tert-butyloxycarbonylarginine (Boc-Arg(Boc)-OPFP) activated with pentafluorophenol is mixed with 2 mg of a fifth-generation symmetrical pentamethine cyanine dye dendrimer G5.

[0078] The characterization and application results of the final prepared transdermal self-assembled nanomedicine are similar to those in Example 1.

[0079] Example 9:

[0080] A method for preparing a transdermal self-assembly nanomedicine, the synthesis steps are basically the same as those in Example 1, except that: in step 3), 10 μL of catalyst diisopropylethylenediamine (DIPEA) is added.

[0081] The characterization and application results of the final prepared transdermal self-assembled nanomedicine are similar to those in Example 1.

[0082] Example 10:

[0083] A method for preparing a transdermal self-assembly nanomedicine, wherein the synthesis steps are substantially the same as those in Example 1, except that: in step 3), 2 mL of trifluoroacetic acid (TFA) is used for deprotection.

[0084] The characterization and application results of the final prepared transdermal self-assembled nanomedicine are similar to those in Example 1.

[0085] Example 11:

[0086] A method for preparing a transdermal self-assembly nanomedicine, the synthesis steps are basically the same as those in Example 1, except that: in step 4), 2 mg of active ester - polyethylene glycol 2000-dibenzocyclooctyne (NHS-PEG2000-DBCO) is added.

[0087] The characterization and application results of the final prepared transdermal self-assembled nanomedicine are similar to those in Example 1.

[0088] Example 12:

[0089] A method for preparing a transdermal self-assembly nanomedicine, the synthesis steps are basically the same as those in Example 1, except that: in step 4), 3 mg of active ester - polyethylene glycol 2000-dibenzocyclooctyne (NHS-PEG2000-DBCO) is added.

[0090] The characterization and application results of the final prepared transdermal self-assembled nanomedicine are similar to those in Example 1.

[0091] Example 13:

[0092] A method for preparing a transdermal self-assembly nanomedicine, wherein the synthesis steps are substantially the same as those in Example 1, except that: in step 5), 4 mg of GR / DBCO is added.

[0093] The characterization and application results of the final prepared transdermal self-assembled nanomedicine are similar to those in Example 1.

Claims

1. A method for preparing a transdermal self-assembling nanomedicine, characterized in that the preparation method comprises: 1) Dissolve 6-MP in a buffer solution, add ZA and PD-1, and stir at room temperature to react to obtain a self-assembled nanodrug 6-MP / Zn / PD-1; 2) 6-MP / Zn / PD-1 was dispersed in a buffer solution, DF-PEG2000-N3 was added, and the mixture was stirred at room temperature to obtain a self-assembled nanodrug 6-MP / Zn / PD-1 / N3 with surface modified azide groups; 3) Boc-Arg(Boc)-OPFP was mixed with G5, a catalyst was added, and the mixture was reacted at room temperature and then deprotected to obtain the arginine-rich dendrimer GR; 4) GR was dissolved in a buffer solution, and NHS-PEG2000-DBCO was added and stirred at room temperature to obtain an arginine-rich dendrimer GR / DBCO with surface modification of dibenzocyclooctyne; 5) Dissolving 6-MP / Zn / PD-1 / N3 in a buffer solution, adding GR / DBCO, and stirring at room temperature to react to obtain a transdermal self-assembled nanodrug 6-MP / Zn / PD-1 / GR; Among them, 6-MP is 6-mercaptopurine, ZA is zinc acetate, PD-1 is programmed death receptor-1, DF-PEG2000-N3 is benzaldehyde-polyethylene glycol 2000-azide group, Boc-Arg(Boc)-OPFP is pentafluorophenol-activated tri-tert-butyloxycarbonyl arginine, G5 is the fifth-generation symmetrical pentamethine cyanine dye dendrimer, and NHS-PEG2000-DBCO is active ester-polyethylene glycol 2000-dibenzocyclooctyne; The usage ratio of 6-MP, ZA and PD-1 in step 1) is 1 mol: 1.25mol: 5-10μmol; The dosage ratio of 6-MP / Zn / PD-1 and DF-PEG2000-N3 in step 2) is 20 mg:1-3 mg; In step 4), the dosage ratio of GR and NHS-PEG2000-DBCO is 10 mg:1-3 mg; The usage ratio of 6-MP / Zn / PD-1 / N3 and GR / DBCO in step 5) is 10 mg:2-5 mg.

2. The method for preparing a transdermal self-assembly nanomedicine according to claim 1, wherein: In step 3), diisopropylethylenediamine (DIPEA) and trifluoroacetic acid (TFA) are used as catalysts for deprotection, and the usage ratio of Boc-Arg(Boc)-OPFP, G5, DIPEA and TFA is 20-80 mg: 2 mg: 3-10 μL: 1-2 mL.

3. A method for preparing a transdermal self-assembled nano drug gel, characterized in that: The transdermal self-assembled nanodrug 6-MP / Zn / PD-1 / GR prepared by the preparation method according to any one of claims 1-2 is mixed with fibrinogen and thrombin to obtain a transdermal self-assembled nanodrug gel Gel@6-MP / Zn / PD-1 / GR.

4. The method for preparing the transdermal self-assembly nano drug gel according to claim 3, wherein: The dosage ratio of the 6-MP / Zn / PD-1 / GR, fibrinogen and thrombin is 2-10 mg:1 mL:1 mL, the thrombin concentration is 2 mg / mL, and the fibrinogen concentration is 10 mg / mL.

5. A transdermal self-assembled nanomedicine obtained according to the preparation method according to any one of claims 1-2.

6. Use of the transdermal self-assembled nanomedicine according to claim 5 in preparing a transdermal drug for treating tumors.

7. A transdermal self-assembled nano drug gel obtained according to the preparation method according to any one of claims 3-4.

8. Use of the transdermal self-assembled nano drug gel according to claim 7 in preparing a drug for transdermal administration to treat tumors.

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