Preparation method of HIV (Human Immunodeficiency Virus) blocking nanoparticles for targeting and activating immune cells
By preparing nanoparticles that target and activate immune cells, the problem of insufficient immune response of HIV virus integration into host cells and the blockade of HIV-1 virus fusion with cells and the removal of infected cells was solved.
Patent Information
- Application Number
- CN202510453216.1
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-04-11
- Publication Date
- 2025-07-04
AI Technical Summary
In the prior art, under combined antiretroviral therapy, HIV virus can still be integrated into host cells to form an infection reservoir, with poor blocking effect and insufficient cellular immune response, which cannot completely remove infected cells.
Nanoparticles targeting and activate immune cells were prepared, and nanoparticles were formed by grafting CD4 and CCR5 antagonist D-Ala-peptide T-amide (DAPTA) and Ce6 photosensitizer onto 4-(4-hydroxymethyl-2-methoxy-5-nitrophenoxy)butyric acid modified silk fibroic protein, and activate macrophages using photodynamics to achieve efficient delivery of CD4 and CCR5 antagonists and blocking HIV-1 virus fusion with cells.
The efficient delivery of CD4 and CCR5 antagonists was achieved, blocking the fusion of HIV-1 virus with cells, and effectively activate macrophages to clear infected immune cells.
Smart Images
Figure CN120242011A_ABST
Abstract
Description
Technical Field
[0001] The present invention belongs to the technical field of biomedical engineering, and particularly relates to a preparation method of nanoparticles for targeting and activating immune cells to block HIV. Background Art
[0002] AIDS, namely acquired immunodeficiency syndrome, is caused by the human immunodeficiency virus HIV and is one of the important public health problems affecting public health. According to the data of the Joint United Nations Programme on HIV / AIDS in 2023, there are approximately 39.9 million people globally infected with the HIV virus. Currently, although the key difficulties in the prevention and early treatment of clinical AIDS are as follows: 1. Under the treatment of combination antiretroviral therapy, the HIV virus can still integrate into host cells to form an infection reservoir, and the blocking effect is not good; 2. The cellular immune response is insufficient to completely eliminate the infected cells.
[0003] HIV-1 infects immune cells, especially CD4+ T lymphocytes and macrophages, which is initiated by the interaction of its surface envelope glycoprotein gp120 with CD4 and a co-receptor on the surface of host cells. Among them, chemokine receptor 5 (CCR5) is the main co-receptor of R5 tropic strains. Therefore, CD4 and CCR5 have also become key targets in the research on blocking HIV-1 infection. CD4 and CCR5 antagonists can effectively prevent and treat infections by blocking the fusion of the HIV-1 virus with cells. However, how to efficiently deliver CD4 and CCR5 antagonists is a key issue in the current therapeutic effect of CD4 and CCR5 antagonists in blocking HIV infection.
[0004] At the same time, regarding how to eliminate the already infected immune cells, activated macrophages can eliminate infected cells in the body. However, patients infected with HIV-1 usually have insufficient macrophage immune response in the body, resulting in the inability to completely eliminate the infected cells. Therefore, how to effectively and controllably activate macrophages is a key issue in the treatment of HIV-1 infection. Summary of the Invention
[0005] Technical problems to be solved:
[0006] In view of the deficiencies of the prior art, the present application solves the technical problems that under the existing combined antiretroviral therapy, the HIV virus can still integrate into host cells to form an infection reservoir, resulting in poor blocking effect; the cellular immune response is insufficient and cannot completely eliminate the infected cells, etc. The present application provides a method for preparing nanoparticles for targeting and activating immune cells to block HIV. In order to improve the delivery effect of CD4 and CCR5 antagonists, CD4 and CCR antagonists (D-Ala-peptide T-amide, DAPTA) and Ce6 photosensitizer are grafted onto silk fibroin modified with 4-(4-hydroxymethyl-2-methoxy-5-nitrophenoxy) butyric acid by light irradiation, and nanoparticles are prepared in methanol to achieve efficient delivery of CD4 and CCR5 antagonists and block the fusion of HIV-1 virus with cells; at the same time, regarding how to effectively and controllably activate macrophages, the present application combines the effect of CD4 and CCR5 antagonistic groups targeting immune cells, and utilizes the photodynamic activation effect of the chlorin e6 group in the nanoparticles on macrophages under near-infrared light irradiation to effectively eliminate the infected immune cells.
[0007] Technical solution:
[0008] To achieve the above object, the present application is realized through the following technical solutions:
[0009] A method for preparing nanoparticles for targeting and activating immune cells to block HIV, comprising the following steps:
[0010] The first step, preparation of silk fibroin modified with 4-(4-hydroxymethyl-2-methoxy-5-nitrophenoxy) butyric acid: Dissolve
[0011] 0.45 g of silk fibroin in 3 - 10 mL of hexafluoroisopropanol, add 0.2 - 2 g of 4-(4-hydroxymethyl-2-methoxy-5-nitrophenoxy) butyric acid, stir in the dark at room temperature until completely dissolved, react for 12 - 48 h, remove the solvent by rotary evaporation, dissolve in water, dialyze for 1 - 3 days, and obtain silk fibroin modified with 4-(4-hydroxymethyl-2-methoxy-5-nitrophenoxy) butyric acid by freeze-drying;
[0012] Step 2. Preparation of silk fibroin loaded with DAPTA and chlorin e6: Dissolve 4-(4-hydroxymethyl-2-methoxy-5-nitrophenoxy)butyric acid modified silk fibroin in ultrapure water to obtain a 0.1% - 1% (w / v) solution of 4-(4-hydroxymethyl-2-methoxy-5-nitrophenoxy)butyric acid modified silk fibroin. Then, add 0.1 - 10 ng of DAPTA and 0.1 mg - 10 mg of chlorin e6 to 10 mL of the 4-(4-hydroxymethyl-2-methoxy-5-nitrophenoxy)butyric acid modified silk fibroin solution. Based on the mild photochemical reaction conditions that avoid drug inactivation, irradiate the silk fibroin solution with blue light at 365 nm - 400 nm and 10 - 100 mW / cm 2 for 1 - 5 min, dialyze in ultrapure water for 1 - 3 days, and obtain 4-(4-hydroxymethyl-2-methoxy-5-nitrophenoxy)butyric acid modified silk fibroin loaded with DAPTA and chlorin e6 by freeze-drying;
[0013] Step 3. Preparation of silk fibroin nanoparticles: Dissolve 4-(4-hydroxymethyl-2-methoxy-5-nitrophenoxy)butyric acid modified silk fibroin loaded with DAPTA and chlorin e6 in ultrapure water to obtain a 1% - 4% (w / v) aqueous silk fibroin solution;
[0014] Drop 10 mL of the aqueous silk fibroin solution into 90 mL of methanol drop by drop, react at room temperature and stir at 50 rpm for 1 hour to obtain a nanoparticle suspension; then, centrifuge at 15000 - 20000 g for 5 - 30 min to obtain nanoparticles, wash with ultrapure water, and freeze-dry to obtain nanoparticles.
[0015] Furthermore, in the first step, dissolve 0.45 g of silk fibroin in 3 mL of hexafluoroisopropanol according to the mass-to-volume ratio, add 2 g of 4-(4-hydroxymethyl-2-methoxy-5-nitrophenoxy)butyric acid, stir in the dark at room temperature until completely dissolved, react for 24 h, remove the solvent by rotary evaporation, dissolve in water, dialyze for 3 days, and obtain 4-(4-hydroxymethyl-2-methoxy-5-nitrophenoxy)butyric acid modified silk fibroin by freeze-drying.
[0016] Furthermore, in the second step, dissolve 4-(4-hydroxymethyl-2-methoxy-5-nitrophenoxy)butyric acid modified silk fibroin in ultrapure water to obtain a 1% (w / v) solution of 4-(4-hydroxymethyl-2-methoxy-5-nitrophenoxy)butyric acid modified silk fibroin.
[0017] Furthermore, in the second step, under blue light at 400 nm and 50 mW / cm 2Irradiate from below for 5 min, dialyze in ultrapure water for 3 days, and obtain silk fibroin modified with 4-(4-hydroxymethyl-2-methoxy-5-nitrophenoxy)butyric acid loaded with DAPTA and chlorin e6 after freeze-drying.
[0018] Furthermore, in the third step, the silk fibroin modified with 4-(4-hydroxymethyl-2-methoxy-5-nitrophenoxy)butyric acid loaded with DAPTA and chlorin e6 is dissolved in ultrapure water to obtain a 4% (w / v) aqueous silk fibroin solution; 10 mL of the aqueous silk fibroin solution is gradually added dropwise into 90 mL of methanol, reacted at room temperature and stirred at 50 rpm for 2 hours to obtain a nanoparticle suspension; then, centrifuged at 18500 g for 15 min to obtain nanoparticles, washed with ultrapure water, and freeze-dried to obtain nanoparticles.
[0019] Beneficial effects:
[0020] The present application provides a method for preparing nanoparticles for targeting and activating immune cells to block HIV. Compared with the prior art, it has the following beneficial effects:
[0021] 1. Utilizing the advantages of fast and mild conditions of the photoreaction, the present application stably grafts CD4 and CCR antagonists (D-Ala-peptide T-amide, DAPTA) and the Ce6 photosensitizer onto silk fibroin modified with 4-(4-hydroxymethyl-2-methoxy-5-nitrophenoxy)butyric acid by light irradiation, and prepares them into nanoparticles to achieve efficient delivery of CD4 and CCR5 antagonists and block the fusion of HIV-1 virus with cells;
[0022] 2. Combining the effect of targeting immune cells with CD4 and CCR5 antagonistic groups, and utilizing the effect of the chlorin e6 group in the nanoparticles to photodynamically activate macrophages under near-infrared light irradiation, the present application not only achieves efficient blocking of the fusion of HIV-1 virus with cells, but also realizes the clearance of infected immune cells. Description of the drawings
[0023] Figure 1 It is a characterization result diagram of the diameter of the nanoparticles for targeting and activating immune cells to block HIV prepared in Example 3 of the present application;
[0024] Figure 2 It is a performance diagram of the photo-activated macrophage immune response of the nanoparticles for targeting and activating immune cells to block HIV of the present application;
[0025] Figure 3 It is a cell compatibility diagram of the nanoparticle product of the present application. Detailed implementation manners
[0026] The present invention will be further described below in conjunction with specific embodiments. It should be understood that these embodiments are only used to illustrate the present invention and not to limit the scope of the present invention. In addition, it should be understood that after reading the content taught by the present invention, those skilled in the art can make various changes or modifications to the present invention, and equivalent changes or modifications also fall within the scope defined by the claims of this application.
[0027] Example 1:
[0028] A preparation method of a targeted and activated immune cell HIV-blocking nanoparticle, comprising the following steps:
[0029] First step, preparation of 4-(4-hydroxymethyl-2-methoxy-5-nitrophenoxy)butyric acid-modified silk fibroin: According to the mass-volume ratio,
[0030] 0.45 g of silk fibroin was dissolved in 3 mL of hexafluoroisopropanol, and 2 g of 4-(4-hydroxymethyl-2-methoxy-5-nitrophenoxy)butyric acid was added. It was stirred in the dark at room temperature until completely dissolved, reacted for 24 h, the solvent was removed by rotary evaporation, and then dissolved in water. After dialysis for 3 days, 4-(4-hydroxymethyl-2-methoxy-5-nitrophenoxy)butyric acid-modified silk fibroin was obtained by freeze-drying;
[0031] Second step, preparation of silk fibroin loaded with DAPTA and chlorin e6: Dissolve 4-(4-hydroxymethyl-2-methoxy-5-nitrophenoxy)butyric acid-modified silk fibroin in ultrapure water to obtain a 1% (w / v) 4-(4-hydroxymethyl-2-methoxy-5-nitrophenoxy)butyric acid-modified silk fibroin solution. Then, add 0.1 ng of DAPTA and 0.1 mg of chlorin e6 to 10 mL of the 4-(4-hydroxymethyl-2-methoxy-5-nitrophenoxy)butyric acid-modified silk fibroin solution; Based on the mild photochemical reaction conditions to avoid drug inactivation, irradiate the silk fibroin solution with 400 nm blue light at 50 mW / cm 2 for 5 min, dialyze in ultrapure water for 3 days, and obtain 4-(4-hydroxymethyl-2-methoxy-5-nitrophenoxy)butyric acid-modified silk fibroin loaded with DAPTA and chlorin e6 by freeze-drying;
[0032] Third step, preparation of silk fibroin nanoparticles: Dissolve 4-(4-hydroxymethyl-2-methoxy-5-nitrophenoxy)butyric acid-modified silk fibroin loaded with DAPTA and chlorin e6 in ultrapure water to obtain a 4% (w / v) aqueous silk fibroin solution; Add 10
[0033] Dropwise add the aqueous solution of silk fibroin in mL into 90 mL of methanol, react at room temperature and stir at 50 rpm for 2 hours to obtain a nanoparticle suspension; then, centrifuge at 18500 g for 15 min to obtain the nanoparticles, wash with ultrapure water, and freeze-dry to obtain the HIV-blocking nanoparticles that target and activate immune cells.
[0034] During application, apply the aqueous solution of nanoparticles at 100 μg / mL to the target position, and irradiate with 660 nm near-infrared light (20
[0035] mW / cm 2 ) for 10 min to activate CD8+ T cells and macrophages in situ at the affected area by photodynamic activation.
[0036] Example 2:
[0037] A preparation method of HIV-blocking nanoparticles that target and activate immune cells, comprising the following steps:
[0038] The first step, preparation of silk fibroin modified with 4-(4-hydroxymethyl-2-methoxy-5-nitrophenoxy)butyric acid: According to the mass-volume ratio,
[0039] Dissolve 0.45 g of silk fibroin in 3 mL of hexafluoroisopropanol, and add 2 g of 4-(4-hydroxymethyl-2-methoxy-5-nitrophenoxy)butyric acid
[0040] Stir in the dark at room temperature until completely dissolved, react for 24 h, remove the solvent by rotary evaporation, dissolve in water, dialyze for 3 days, and obtain silk fibroin modified with 4-(4-hydroxymethyl-2-methoxy-5-nitrophenoxy)butyric acid by freeze-drying;
[0041] The second step, preparation of silk fibroin loaded with DAPTA and chlorin e6: Dissolve the silk fibroin modified with 4-(4-hydroxymethyl-2-methoxy-5-nitrophenoxy)butyric acid in ultrapure water to obtain a 1% (w / v) aqueous solution of silk fibroin modified with 4-(4-hydroxymethyl-2-methoxy-5-nitrophenoxy)butyric acid, and then add 0.1 ng of DAPTA and 10 mg of chlorin e6 to 10 mL of the aqueous solution of silk fibroin modified with 4-(4-hydroxymethyl-2-methoxy-5-nitrophenoxy)butyric acid; Based on the mild photochemical reaction conditions to avoid drug inactivation, irradiate the aqueous solution of silk fibroin with 400 nm blue light at 50 mW / cm 2 for 5 min, dialyze in ultrapure water for 3 days, and obtain silk fibroin modified with 4-(4-hydroxymethyl-2-methoxy-5-nitrophenoxy)butyric acid loaded with DAPTA and chlorin e6 by freeze-drying;
[0042] The third step is the preparation of silk fibroin nanoparticles: dissolve the 4-(4-hydroxymethyl-2-methoxy-5-nitrophenoxy)butyric acid-modified silk fibroin loaded with DAPTA and dihydrochlorin e6 in ultrapure water to obtain a 4% (w / v) silk fibroin aqueous solution; drip 10 mL of the silk fibroin aqueous solution dropwise into 90 mL of methanol, react at room temperature and stir at 50 rpm for 2 hours to obtain a nanoparticle suspension; then, centrifuge at 18500 g for 15 min to obtain nanoparticles, wash with ultrapure water, and freeze-dry to obtain HIV blocking nanoparticles that target and activate immune cells.
[0043] Embodiment 3:
[0044] A method for preparing HIV blocking nanoparticles that target and activate immune cells comprises the following steps:
[0045] The first step is the preparation of 4-(4-hydroxymethyl-2-methoxy-5-nitrophenoxy)butyric acid-modified silk fibroin:
[0046] 0.45 g of silk fibroin was dissolved in 3 mL of hexafluoroisopropanol, and 2 mL of 4-(4-hydroxymethyl-2-methoxy-5-nitrophenoxy)butyric acid was added.
[0047] g, stirred at room temperature in the dark until completely dissolved, reacted for 24 h, the solvent was removed by rotary evaporation, and then dissolved in water, dialyzed for 3 days, and lyophilized to obtain 4-(4-hydroxymethyl-2-methoxy-5-nitrophenoxy)butyric acid-modified silk fibroin;
[0048] The second step is the preparation of silk fibroin loaded with DAPTA and dihydrochlorin e6: the silk fibroin modified with 4-(4-hydroxymethyl-2-methoxy-5-nitrophenoxy)butyric acid was dissolved in ultrapure water to obtain a 1% (w / v) 4-(4-hydroxymethyl-2-methoxy-5-nitrophenoxy)butyric acid modified silk fibroin solution, and then 10 ng DAPTA and 10 mg dihydrochlorin e6 were added to 10 mL of the 4-(4-hydroxymethyl-2-methoxy-5-nitrophenoxy)butyric acid modified silk fibroin solution; based on the characteristics of mild photochemical reaction conditions to avoid drug inactivation, the silk fibroin solution was irradiated with 400 nm blue light 50 mW / cm 2 The mixture was irradiated for 5 min, dialyzed in ultrapure water for 3 days, and freeze-dried to obtain 4-(4-hydroxymethyl-2-methoxy-5-nitrophenoxy)butyric acid-modified silk fibroin loaded with DAPTA and dihydrochlorin e6;
[0049] Step 3: Preparation of silk fibroin nanoparticles: Dissolve silk fibroin modified with 4-(4-hydroxymethyl-2-methoxy-5-nitrophenoxy)butyric acid loaded with DAPTA and chlorin e6 in ultrapure water to obtain a 4% (w / v) aqueous silk fibroin solution; drop 10 mL of the aqueous silk fibroin solution into 90 mL of methanol drop by drop, react at room temperature and stir at 50 rpm for 2 hours to obtain a nanoparticle suspension; then, centrifuge at 18500 g for 15 min to obtain nanoparticles, wash with ultrapure water, and freeze-dry to obtain nanoparticles for targeting and activating immune cells to block HIV.
[0050] Test Example 1:
[0051] Characterize the diameter of the nanoparticles for targeting and activating immune cells to block HIV prepared in Example 3 using a laser scattering microscope. As Figure 1 shown, the diameter of the obtained nanoparticles is 138.3 ± 1.3 μm.
[0052] Test Example 2:
[0053] Use in vitro cell culture to test the anti-HIV virus cell fusion effect of the product. Seed TZM-b1 cells in a 24-well plate. Incubate the nanoparticles prepared in Examples 1, 2, and 3 at 100 μg / mL with HIV pseudovirus at a concentration of 15 ng / μL and TZM-bl cells for 48 h, then fix, stain, and count. As shown in Table 1, the nanoparticle product prepared in Example 3 has an HIV-1 virus cell fusion blocking rate exceeding 80%, which is significantly higher than the nanoparticles prepared in Examples 1 and 2, proving that the nanoparticle product prepared in Example 3 has a good anti-HIV virus cell fusion effect.
[0054] Table 1 Blocking virus cell fusion effect of nanoparticles for targeting and activating immune cells to block HIV
[0055]
[0056]
[0057] Test Example 3:
[0058] Use a mouse dorsal skin injury model to test the performance of the product in activating macrophages at the affected area under near-infrared light. Apply an aqueous solution of the nanoparticles prepared in Example 3 at 100 μg / mL to the affected area, and irradiate with 660 nm near-infrared light (20 mW / cm 2 ) for 10 min. After 24 hours, take tissue from the affected area for immunofluorescence staining to observe the activation of immune cells, and compare with the tissue of untreated mice. As Figure 2As shown, the activated macrophages are marked by red fluorescent iNOS. After treatment, the macrophages at the wound site are significantly activated, demonstrating that the product has good performance in photoactivating macrophage immune response.
[0059] Test Example 4
[0060] The cytotoxicity of the product was tested using in vitro cell culture. L929 cells were seeded in 24-well plates and cultured in a medium containing 100 μg / mL of the nanoparticles prepared in Example 3 for 3 days. The cell viability was detected using the Cell Counting Kit-8 assay. As Figure 3 shown, after 3 days of culture, the survival rate of L929 cells on the product exceeded 95%, demonstrating that the nanoparticle product has good cell compatibility.
[0061] The above provides an exemplary description of the present invention. It should be noted that any simple deformation, modification, or equivalent substitution that can be made by those skilled in the art without creative efforts falls within the protection scope of the present invention without departing from the core of the present invention.
Claims
1. A preparation method of a targeted and immunocyte-activated HIV-blocking nanoparticle, characterized in that It includes the following steps: The first step, preparation of 4-(4-hydroxymethyl-2-methoxy-5-nitrophenoxy)butyric acid-modified silk fibroin: Dissolve 0.45 g of silk fibroin in 3 - 10 mL of hexafluoroisopropanol according to the mass-volume ratio, and add 0.2 - 2 g of 4-(4-hydroxymethyl-2-methoxy-5-nitrophenoxy)butyric acid. Stir in the dark at room temperature until completely dissolved, react for 12 - 48 h, remove the solvent by rotary evaporation, dissolve in water, dialyze for 1 - 3 days, and obtain 4-(4-hydroxymethyl-2-methoxy-5-nitrophenoxy)butyric acid-modified silk fibroin by freeze-drying; Step 2, Preparation of silk fibroin loaded with DAPTA and chlorin e6: Dissolve 4-(4-hydroxymethyl-2-methoxy-5-nitrophenoxy)butyric acid-modified silk fibroin in ultrapure water to obtain a 0.1% - 1% (w / v) 4-(4-hydroxymethyl-2-methoxy-5-nitrophenoxy)butyric acid-modified silk fibroin solution. Then, add 0.1 - 10 ng of DAPTA and 0.1 mg - 10 mg of chlorin e6 to 10 mL of the 4-(4-hydroxymethyl-2-methoxy-5-nitrophenoxy)butyric acid-modified silk fibroin solution. Based on the mild photochemical reaction conditions that avoid drug inactivation, irradiate the silk fibroin solution with blue light at 365 nm - 400 nm and 10 - 100 mW / cm 2 for 1 - 5 min, dialyze it in ultrapure water for 1 - 3 days, and lyophilize it to obtain 4-(4-hydroxymethyl-2-methoxy-5-nitrophenoxy)butyric acid-modified silk fibroin loaded with DAPTA and chlorin e6; The third step, preparation of silk fibroin nanoparticles: Dissolve 4-(4-hydroxymethyl-2-methoxy-5-nitrophenoxy)butyric acid-modified silk fibroin loaded with DAPTA and chlorin e6 in ultrapure water to obtain a 1% - 4% (w / v) aqueous silk fibroin solution; Drop 10 mL of the aqueous silk fibroin solution into 90 mL of methanol drop by drop, react at room temperature and stir at 50 rpm for 1 hour to obtain a nanoparticle suspension; Then, centrifuge at 15000 - 20000 g for 5 - 30 min to obtain nanoparticles, wash with ultrapure water, and freeze-dry to obtain nanoparticles.
2. The preparation method of the HIV-blocking nanoparticles targeting and activating immune cells according to claim 1, wherein: In the first step, 0.45 g of silk fibroin is dissolved in 3 mL of hexafluoroisopropanol according to the mass-volume ratio, 2 g of 4-(4-hydroxymethyl-2-methoxy-5-nitrophenoxy)butyric acid is added, stirred in the dark at room temperature until completely dissolved, reacted for 24 h, the solvent is removed by rotary evaporation, dissolved in water, dialyzed for 3 days, and 4-(4-hydroxymethyl-2-methoxy-5-nitrophenoxy)butyric acid-modified silk fibroin is obtained by freeze-drying.
3. The preparation method of the HIV-blocking nanoparticles targeting and activating immune cells according to claim 1, wherein: In the second step, 4-(4-hydroxymethyl-2-methoxy-5-nitrophenoxy)butyric acid-modified silk fibroin is dissolved in ultrapure water to obtain a 1% (w / v) 4-(4-hydroxymethyl-2-methoxy-5-nitrophenoxy)butyric acid-modified silk fibroin solution.
4. The preparation method of the HIV-blocking nanoparticles targeting and activating immune cells according to claim 1, wherein: In the second step, under irradiation with 50 mW / cm² of 400-nm blue light 2 for 5 min, dialysis is carried out in ultrapure water for 3 days, and freeze-drying is performed to obtain silk fibroin modified with 4-(4-hydroxymethyl-2-methoxy-5-nitrophenoxy)butyric acid loaded with DAPTA and chlorin e6.
5. The preparation method of the HIV-blocking nanoparticles targeting and activating immune cells according to claim 1, wherein: In the third step, 4-(4-hydroxymethyl-2-methoxy-5-nitrophenoxy)butyric acid-modified silk fibroin loaded with DAPTA and chlorin e6 is dissolved in ultrapure water to obtain a 4% (w / v) aqueous silk fibroin solution; Drop 10 mL of the aqueous silk fibroin solution into 90 mL of methanol drop by drop, react at room temperature and stir at 50 rpm for 2 hours to obtain a nanoparticle suspension; Then, centrifuge at 18500 g for 15 min to obtain nanoparticles, wash with ultrapure water, and freeze-dry to obtain nanoparticles.