A compound and its preparation method, a modular polypeptide nanovaccine and its preparation method and application

Abstract

This invention relates to a compound and its preparation method, a modular peptide nanovaccine and its preparation method and application, and relates to the pharmaceutical field. The chemical structural formula of the compound is shown in Formula I. This invention provides a compound containing an mPEG-PLA structure, a protoporphyrin structure chelated with cobalt ions, and a β-cyclodextrin structure, endowing it with excellent binding ability to peptides and excellent loading capacity with adjuvants, achieving co-delivery of peptides and adjuvants. Based on this, this invention also provides a modular peptide nanovaccine with high peptide loading rate and "plug-and-play" characteristics. It can significantly inhibit tumor growth in a B16 subcutaneous tumor model, providing an efficient and flexible technical platform for the prevention of melanoma and other tumors and the clinical translation of peptide vaccines, with broad application prospects.

Description

Technical Field

[0001] This invention relates to the pharmaceutical field, and more particularly to a compound and its preparation method, a modular polypeptide nanovaccine and its preparation method and application. Background Technology

[0002] Vaccines play a crucial role in the prevention and treatment of diseases. With the continuous development of biotechnology and materials science, the development of novel, highly effective, safe, and customizable vaccines has become a research hotspot. Peptide vaccines, due to their high specificity and minimal side effects, show broad application prospects in areas such as cancer treatment and infectious disease prevention. However, peptides themselves suffer from poor stability, weak immunogenicity, and low delivery efficiency, limiting their widespread clinical application. Therefore, improving the peptide loading capacity and designing modular nanoparticle vaccine carriers have become critical issues that urgently need to be addressed. Summary of the Invention

[0003] To address the above problems, this invention provides a compound and its preparation method, a modular polypeptide nanovaccine and its preparation method and application.

[0004] In a first aspect, the present invention provides a compound having the chemical structural formula shown in Formula I;

[0005] Equation I: ;

[0006] Where m can take values ​​from 7 to 28, for example, 7, 15, 20, 28, etc.

[0007] The value of n is 35 to 110, for example, it can be 35, 40, 60, 85, 100, 110, etc.

[0008] Secondly, the present invention provides a method for preparing the compound according to any one of the first aspects, the method comprising the following steps:

[0009] A ring-opening polymerization reaction was carried out using polyethylene glycol monomethyl ether and lactide as raw materials to obtain mPEG-PLA block polymer;

[0010] The mPEG-PLA block polymer and pre-activated carboxyl protoporphyrin were esterified, and then a cobalt source was added for chelation to obtain the mPEG-PLA-PPIX-COOH intermediate.

[0011] The mPEG-PLA-PPIX-COOH intermediate and β-cyclodextrin were subjected to an amidation reaction to obtain the compound.

[0012] Furthermore, the conditions and parameters of the ring-opening polymerization reaction include: the catalyst includes stannous octoate, the molar ratio of polyethylene glycol monomethyl ether to lactide is 1:(10-30), the reaction temperature is 110-130℃, and the reaction time is 24-48h;

[0013] And / or, the number-average molecular weight of the polyethylene glycol monomethyl ether is 500~2000 g / mol;

[0014] And / or, the number-average molecular weight of the mPEG-PLA block polymer is 3000-10000 g / mol;

[0015] And / or, by mass percentage, the hydrophilic mPEG segment in the mPEG-PLA block polymer accounts for 10%-30%;

[0016] And / or, the pre-activated carboxyl protoporphyrin is activated using 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and 4-dimethylaminopyridine as activators, wherein the molar ratio of 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, 4-dimethylaminopyridine and protoporphyrin is (2.0-2.5):(2.2-2.8):1;

[0017] And / or, the conditional parameters of the chelation reaction include: the cobalt source includes anhydrous cobalt chloride, Co 2+ The molar ratio of the porphyrin to the protoporphyrin was (8-12):1, the reaction temperature was 25℃, and the reaction time was 12 h.

[0018] And / or, the conditions and parameters of the amidation reaction include: using benzotriazole-N,N,N',N'-tetramethylurea hexafluorophosphate and N,N-diisopropylethylamine as the coupling system, wherein the molar ratio of benzotriazole-N,N,N',N'-tetramethylurea hexafluorophosphate, N,N-diisopropylethylamine and β-cyclodextrin is (1.2-1.5):(3.5-4.5):1, the reaction temperature is 25℃, and the reaction time is 48 h.

[0019] Thirdly, the present invention provides a modular peptide nanovaccine, comprising:

[0020] carrier;

[0021] The polypeptide and adjuvant loaded on the carrier;

[0022] The carrier is a nanoparticle formed by the self-assembly of the compound described in the first aspect or the compound prepared by the preparation method described in the second aspect in an aqueous phase.

[0023] Furthermore, the average particle size of the nanoparticles is 50-200 nm;

[0024] And / or, the coefficient of variation of the dispersibility of the nanoparticles is ≤0.3.

[0025] Furthermore, the polypeptide includes a His-tagged antigen polypeptide, which includes a TYRP-1 polypeptide;

[0026] And / or, the loading rate of the peptide is ≥95%, specifically referring to the loading rate of His-tagged antigen peptides, such as TYRP-1 peptide and TATK peptide.

[0027] Furthermore, the adjuvant includes R848;

[0028] And / or, the adjuvant is loaded into nanoparticles through the hydrophobic cavity of β-cyclodextrin in the compound, and the loading rate of the adjuvant is ≥70%.

[0029] Fourthly, the present invention provides a method for preparing the modular peptide nanovaccine described in the third aspect, the method comprising the following steps:

[0030] The compound was dissolved in an organic solvent and then added dropwise to a PBS solution. The mixture was then stirred to evaporate the organic solvent and purified by a first filtration to obtain a nanoparticle solution.

[0031] The nanoparticle solution was incubated with peptides and adjuvants, followed by a second filtration to obtain the modular peptide nanovaccine.

[0032] Further, the organic solvent includes at least one of tetrahydrofuran, methanol, and ethanol, and the concentration of the compound in the organic solvent is 2.0-3.0 mg / mL; the dropping rate is 45-55 μL / min, the stirring temperature is 37-40℃, and the stirring time is 12 h; the first filtration uses a 0.22 μm polyethersulfone membrane, and the second filtration uses an ultrafiltration tube with a molecular weight cutoff of 100 kDa;

[0033] And / or, the mass ratio of the polypeptide to the nanoparticles is (0.1-0.2):1, and the mass ratio of the adjuvant to the nanoparticles is (0.05-0.1):1;

[0034] And / or, the incubation conditions include: incubation at room temperature for 1.5 to 2.5 hours or incubation at 4°C for 10 to 14 hours.

[0035] Fifthly, the present invention provides the application of the modular polypeptide nanovaccine described in the third aspect or the modular polypeptide nanovaccine prepared by the preparation method described in the fourth aspect in the preparation of drugs for the prevention or treatment of tumors.

[0036] The technical solutions provided in the embodiments of the present invention have at least the following advantages compared with the prior art:

[0037] This invention provides a compound and its preparation method, a modular peptide nanovaccine and its preparation method, and its application. The compound contains an mPEG-PLA structure, a protoporphyrin structure chelated with cobalt ions, and a β-cyclodextrin structure, endowing it with excellent binding ability to peptides and excellent loading capacity with adjuvants, achieving co-delivery of peptides and adjuvants. Based on this, this invention also provides a modular peptide nanovaccine with high peptide loading rate and "plug-and-play" characteristics. It can significantly inhibit tumor growth in a B16 subcutaneous tumor model, providing an efficient and flexible technical platform for the prevention of melanoma and other tumors and the clinical translation of peptide vaccines, with broad application prospects. Specifically:

[0038] (1) Balancing high peptide loading and modular design

[0039] via PPIX-Co 2+ (Protoporphyrin-Co) 2+ The specific binding of the complex to the His (histidine) tag significantly improves the peptide loading rate. At the same time, by replacing antigenic peptides with different His tags (such as specific antigenic peptides for different tumors), "plug-and-play" modular preparation can be achieved without redesigning the carrier, greatly shortening the vaccine development cycle and making it suitable for the prevention of a variety of diseases.

[0040] (2) Antigen-adjuvant co-delivery enhances immune response

[0041] The compound provided in this invention serves as a support (denoted as mPEG-PLA-PPIX (Co)). 2+ The β-CD (β-cyclodextrin) hydrophobic cavity in its structure can simultaneously encapsulate adjuvants such as R848, achieving synergistic delivery of antigen and adjuvant, avoiding the problems of adjuvant loss or asynchronous delivery in traditional vaccines; R848 can activate dendritic cells, promote antigen presentation, thereby enhancing the immunogenicity of peptide vaccines and improving the tumor prevention effect.

[0042] (3) Good biocompatibility and safety

[0043] The compound provided in this invention serves as a carrier, with the carrier raw material mPEG-PLA block polymer (polyethylene glycol monomethyl ether-polylactic acid block polymer) being a commonly used biodegradable material in clinical practice. PPIX (protoporphyrin), β-CD, and low-dose Co... 2+ All vaccines exhibit good biocompatibility and no obvious toxic side effects. In mouse experiments, the weight of mice in the vaccine group remained stable, and no obvious adverse reactions were observed, providing a safe basis for clinical translation.

[0044] (4) The preparation process is simple and can be mass-produced.

[0045] The raw materials used in this invention are readily available, and the synthesis steps (such as polymerization, coupling, and chelation) are all conventional polymer synthesis techniques that do not require special equipment. The nano-assembly process is achieved through solvent displacement, which is simple to operate, highly reproducible, and easy to scale up for industrial production.

[0046] (5) Clear tumor prevention effect

[0047] In the B16 tumor model, this nanovaccine significantly inhibited tumor growth and prolonged mouse survival time, demonstrating its clear therapeutic value. Furthermore, by adjusting the types of antigenic peptides, it can be extended to the prevention of other tumors or infectious diseases, showing broad application prospects. Attached Figure Description

[0048] The accompanying drawings, which are incorporated in and form part of this specification, illustrate embodiments consistent with the invention and, together with the description, serve to explain the principles of the invention.

[0049] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, for those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0050] Figure 1 The synthesis process of the mPEG-PLA block polymer in Example 1 of this invention is shown below.

[0051] Figure 2 This is the 1H NMR spectrum of the mPEG-PLA block polymer in Example 1 of the present invention.

[0052] Figure 3 [mPEG-PLA]2-PPIX (Co) in Embodiment 1 of the present invention 2+ The molecular structure diagram of ).

[0053] Figure 4 The mPEG-PLA-PPIX (Co) in Example 1 of this invention 2+ The proton NMR spectrum of βCD.

[0054] Figure 5 This is a transmission electron microscope image of the modular nanovaccine in Example 2 of the present invention.

[0055] Figure 6 This is a line graph showing the construction of the B16 tumor prevention model based on the TYRP-1 antigen peptide and the tumor volume in Example 2 of the present invention. Detailed Implementation

[0056] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0057] Unless otherwise specified, all raw materials, reagents, instruments and equipment used in this invention can be purchased from the market or prepared by existing methods.

[0058] The present invention will be further illustrated below with reference to specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Experimental methods in the following embodiments, unless otherwise specified, are generally performed according to national standards. If no corresponding national standard exists, then generally accepted international standards, conventional conditions, or conditions recommended by the manufacturer are followed.

[0059] The main raw material information involved in the following examples is as follows:

[0060] TYRP-1 polypeptide (or TYRP-1 antigenic peptide): synthesized by Suzhou Modifu Biotechnology, the specific amino acid sequence of which is represented as shown in SEQ ID NO: 1 with a 6His tag, SEQ ID NO: 1 as shown below: CTAPDNLGYM.

[0061] TATK peptide: synthesized by Suzhou Modifu Biotechnology, its specific amino acid sequence is as shown in SEQ ID NO: 2 with a 6His tag, SEQ ID NO: 2 is as follows: YARKAARQARA; as a membrane-penetrating peptide, its role and function is to enhance the uptake and utilization of antigens.

[0062] Example 1

[0063] This example provides a compound with the following chemical structural formula:

[0064] ;

[0065] Where m ranges from 7 to 28, and n ranges from 35 to 110.

[0066] The preparation method of the above-mentioned compound includes the following steps:

[0067] Step 1: [mPEG-PLA]2-PPIX (Co 2+ The preparation of the polymer units is shown below:

[0068] Step 1.1: As Figure 1 As shown, the preparation method of mPEG-PLA block polymer includes:

[0069] Weigh 1.0 g (7.0 mmol) of lactide and place it in a dry, anhydrous Schlenk tube (dry under vacuum at 65 °C for 24 h, then purge with nitrogen to restore atmospheric pressure), add 5 mL of anhydrous toluene; add the starting material mPEG2000 (1.0 g); add the catalyst 3.18 μmol (1.29 mg, 0.02 g / mL, 64.4 μL) of stannous octoate; stir the reaction in an oil bath at 110 °C for 48 h; after the reaction is complete, precipitate the product three times with 50 mL of n-hexane; and dry under vacuum to obtain the final product.

[0070] Step 1.2: The preparation of protoporphyrin (PPIX) is shown below:

[0071] 3.2598 g (5 mmol) of heme chloride and 150 mL of formic acid were added to a 500 mL three-necked flask and stirred until dissolved. The mixture was slowly heated to 110 °C in an oil bath. When reflux occurred, 1.4 g of reducing Fe powder was added in three batches over 10 min. After the addition was complete, the mixture was refluxed for 1 h. After the reaction was complete, the reaction system was cooled to room temperature. Unreacted Fe powder was removed by filtration, and the precipitate was washed three times with a small amount of formic acid. The collected filtrate was poured into a beaker containing distilled water, and 6 mol / L NaOH solution (7.2 g NaOH in 30 mL UP water) was slowly added dropwise. The pH of the filtrate was adjusted to between 4 and 5 to precipitate the product. The solution was allowed to stand for 2 h, and the precipitate was collected by filtration using a Buchner funnel and washed three times with deionized water. The precipitate was dried in a vacuum oven, and the product was recrystallized from dichloromethane to obtain 1.6816 g of pure protoporphyrin, with a yield of 59.77%.

[0072] Step 1.3: [mPEG-PLA]2-PPIX (Co 2+ The preparation of ) is as follows:

[0073] Protoporphyrin (PPIX) (168.8 mg, 0.3 mmol) was dissolved in 24 mL of anhydrous DMF. 1-Ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC) (138.2 mg, 0.72 mmol) (soluble in water) was dissolved in 4 mL of anhydrous DCM and added to the reaction mixture. 4-Dimethylaminopyridine (DMAP) (97.8 mg, 0.8 mmol) was dissolved in 4 mL of anhydrous DCM and added to the reaction mixture. The mixture was stirred in an ice-water bath for 30 min. The mPEG-PLA block polymer (0.6 mmol) synthesized in step one of Example 1 was dissolved in 8 mL of anhydrous DCM and added to the pre-cooled mixture. The reaction was carried out under nitrogen protection in an ice-water bath for 48 h. After the reaction was complete, the DCM was removed by rotary evaporation under reduced pressure, and the mixture was precipitated with 500 mL of UP water, filtered, and dried. The dried solid powder was thoroughly dissolved in 10 mL of DCM, and 389.5 mg (3 mmol) of anhydrous cobalt chloride (dissolved in 2 mL of methanol solution) was added to chelate Co. 2 + The reaction was stirred at 25°C for 12 h. After the reaction was complete, the solvent was removed by rotary evaporation under reduced pressure, and the mixture was washed with 500 mL of UP water, filtered, and dried. The target product was obtained by silica gel column chromatography.

[0074] Step 2: mPEG-PLA-PPIX (Co 2+ The synthesis of the βCD polymer unit is shown below:

[0075] Step 2.1: The preparation of mPEG-PLA-PPIX-COOH is shown below:

[0076] The synthesized protoporphyrin (PPIX) (168.8 mg, 0.3 mmol) was dissolved in 24 mL of anhydrous N,N-dimethylformamide (DMF). 1-Ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC) (138.2 mg, 0.72 mmol) was dissolved in 4 mL of anhydrous dichloromethane (DCM) and added to the reaction mixture. 4-Dimethylaminopyridine (DMAP) (97.8 mg, 0.8 mmol) was dissolved in 4 mL of anhydrous DCM and added to the reaction mixture. The mixture was stirred in an ice-water bath for 30 min. The synthesized mPEG-PLA block polymer (0.24 mmol) was dissolved in 8 mL of anhydrous DMF and added to the pre-cooled mixture. Under nitrogen protection, the reaction was carried out in an ice-water bath for 12 h, followed by a further 12 h at room temperature. After the reaction was complete, DCM was removed by rotary evaporation under reduced pressure, and the product was precipitated with 500 mL of UP water, filtered, and dried. The target product was obtained by silica gel column chromatography.

[0077] Step 2.2: mPEG-PLA-PPIX (Co2+ The preparation of βCD is shown below:

[0078] mPEG-PDLLA-PPIX (100 mg, 50 μmol) was dissolved in 5 mL of anhydrous DMF. Benzotriazole-N,N,N',N'-tetramethylurea hexafluorophosphate (HBTU) (28.4 mg, 75 μmol) was dissolved in 1 mL of anhydrous DMF and added to the reaction mixture. N,N-diisopropylethylamine (DIPEA) (27.1 mg, 210 μmol) was dissolved in 1 mL of anhydrous DMF and added to the reaction mixture. The mixture was stirred at room temperature for 30 min. An amino compound (60 μmol, specifically mono-(6-hexamethylenediamino-6-deoxy)-β-cyclodextrin) was dissolved in 2 mL of anhydrous DMF and added to the above mixture. The reaction was carried out under nitrogen atmosphere for 48 h. 500 mg of anhydrous cobalt chloride was completely dissolved in 5 mL of DMF solution, cooled, and then added to the above reaction mixture. The reaction was carried out under nitrogen atmosphere with stirring at room temperature for 12 h. After the reaction was complete, DCM was removed by rotary evaporation under reduced pressure, the product was dialyzed against UP water for four days, and then lyophilized to obtain mPEG-PLA-PPIX (Co 2+ )-βCD.

[0079] The 1H NMR spectrum of the mPEG-PLA block polymer in this example is as follows: Figure 2 As shown, [mPEG-PLA]2-PPIX(Co 2+ The molecular structure diagram of ) is as follows Figure 3 As shown, mPEG-PLA-PPIX (Co 2+ The proton NMR spectrum of βCD is shown below. Figure 4 As shown.

[0080] Example 2

[0081] This example provides a modular peptide nanovaccine, comprising:

[0082] carrier;

[0083] The polypeptide and adjuvant loaded on the carrier;

[0084] The polypeptide is a His-tagged antigen polypeptide, the adjuvant is R848, and the carrier is the compound in Example 1 (i.e., mPEG-PLA-PPIX (Co 2+ Nanoparticles formed by the self-assembly of βCD in an aqueous phase.

[0085] The preparation method of the above-mentioned modular polypeptide nanovaccine includes the following steps:

[0086] Weigh 10.0 mg of mPEG-PDLLA-PPIX (Co 2+β-CD was dissolved in 4 mL of tetrahydrofuran (THF) to a concentration of 2.5 mg / mL, and 2.0 mL was taken. The prepared solution (total 2.5 mL) was mixed and drawn into a 5 mL syringe. Using a syringe pump, the solution was added to 15 mL of PBS*2 at a rate of 50 μL / min. The mixture was magnetically stirred at 40°C for 12 h, and the organic solvent was evaporated with the syringe open. The evaporated solution was filtered through a 0.22 μm filter membrane (Millipore Express PES membrane Filter Unit), and then transferred to a 100 kDa ultrafiltration tube and centrifuged (3000 rpm / min; 15 min). For the first use of the ultrafiltration tube, it should be soaked in pure water for 1 h beforehand. It can be reused; after each use, the tube should be filled with pure water to maintain purity. Centrifugation was performed using a horizontal rotor, with the membrane in the ultrafiltration tube perpendicular to the central rotor. After centrifugation, the lower layer was collected, and the eluted solution was defined as a 5-unit nanoparticle solution, which was then diluted to 4 mL. Five units of nanoparticles (4 mL) were added to 1.5 mg TYRP-1 peptide (150 μL, 10 mg / mL), 0.75 mg TATK peptide (75 μL, 10 mg / mL), and 750 μL R848 (5 mg dissolved in 5 mL PBS, 1 mg / mL), bringing the volume to 5 mL. The mixture was incubated at room temperature for 2 h or overnight at 4°C. After incubation, the mixture was transferred to a 5 mL (100 kDa) ultrafiltration tube and centrifuged (3000 rpm / min; 15 min). The lower and upper layers were collected; this solution was defined as the 5-unit nanovaccine. The concentration of R848 in the lower layer was detected by fluorescence, and the concentration of the loaded peptide was determined by BCA assay. The peptide concentration was then adjusted to 1 mg / mL. The concentration of R848 in the supernatant was detected by fluorescence detection, and the concentration of loaded peptides was detected by BCA kit (5 samples in total: upper and lower layers of experimental group, upper and lower layers of control group, and 0.25 mg / mL peptide standard). The loading rates of R848 and peptides in the supernatant were calculated by combining the results.

[0087] Testing revealed that the modular peptide nanovaccine obtained in this example exhibited a coefficient of variation for dispersibility of <0.3 and an average particle size of 50-200 nm. The peptide loading rate was >95%, and the adjuvant loading rate was >60%. Furthermore, the transmission electron microscopy (TEM) image of the modular peptide nanovaccine obtained in this example is shown below. Figure 5 As shown, this indicates that the particle size meets the design requirements and has good uniformity.

[0088] Test case

[0089] This example evaluates the antitumor therapeutic effect of the modular polypeptide nanovaccine obtained in Example 2 above.

[0090] The testing method is as follows: Male C57BL / 6 mice (16-18 g) aged approximately 6-8 weeks were shaved from the base of their tails beforehand. The mice were divided into two groups (n=8 per group): a PBS group and a PPIX NPs + peptide + R848 (adjuvant) group. Each group was subcutaneously injected with 100 μL of vaccine, containing 100 μg / mouse of peptide (specifically TYRP-1 peptide + TATK peptide) and 10 μg / mouse of R848. Immunization was repeated twice, 7 days apart. Fourteen days later, the right back of the mice was shaved beforehand. After isoflurane anesthesia, the right back of the mice was exposed, and 500,000 B16 cells were subcutaneously inoculated. Tumor growth was observed, and tumor volume was recorded daily using calipers. Tumor volume = (tumor long diameter * tumor short diameter) 2 ) / 2. When the tumor volume grows to 1000 mm 3 Mice were observed to die at the designated time. Tumor growth curves, survival rates, and weight changes in each group were recorded. Serum markers, CD4, CD8, Treg, lymph node / tumor tissue sections, dendritic cells (DCs), and cytokines were measured in mice injected with the PPIX nanovaccine in the B16 prevention model.

[0091] Test results are as follows Figure 6 As shown, by Figure 6 It can be seen that the designed and synthesized nanovaccines showed good tumor prevention effects in the B16-OVA mouse prevention model.

[0092] Various embodiments of the present invention may exist in the form of a range; it should be understood that the description in the form of a range is merely for convenience and brevity and should not be construed as a hard limitation on the scope of the invention; therefore, it should be considered that the range description has specifically disclosed all possible subranges and single numerical values ​​within that range. For example, it should be considered that the range description from 1 to 6 has specifically disclosed subranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6, etc., and single numbers within the range, such as 1, 2, 3, 4, 5, and 6, regardless of the range. Furthermore, whenever a numerical range is referred to herein, it means including any referenced number (fraction or integer) within the range referred to.

[0093] The above description is merely a specific embodiment of the present invention, enabling those skilled in the art to understand or implement the invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the invention. Therefore, the present invention is not to be limited to the embodiments shown herein, but is to be accorded the widest scope consistent with the principles and novel features claimed herein.

Claims

1. A compound, characterized in that, The chemical structural formula of the compound is shown in Formula I; Equation I: ; Where m takes the value of 42 and n takes the value of 28.

2. A method for preparing the compound according to claim 1, characterized in that, The method for preparing the compound includes the following steps: A ring-opening polymerization reaction was carried out using polyethylene glycol monomethyl ether and lactide as raw materials to obtain mPEG-PLA block polymer; The mPEG-PLA block polymer and pre-activated carboxyl protoporphyrin were esterified, and then a cobalt source was added for chelation to obtain the mPEG-PLA-PPIX-COOH intermediate. The mPEG-PLA-PPIX-COOH intermediate and mono-(6-hexanediamine-6-deoxy)-β-cyclodextrin were subjected to an amidation reaction to obtain the compound.

3. The method for preparing the compound according to claim 2, characterized in that, The conditions and parameters for the ring-opening polymerization reaction include: the catalyst is stannous octoate, the molar ratio of polyethylene glycol monomethyl ether to lactide is 1:(10-30), the reaction temperature is 110-130℃, and the reaction time is 24-48 h. And / or, the pre-activated carboxyl protoporphyrin is activated using 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and 4-dimethylaminopyridine as activators, wherein the molar ratio of 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, 4-dimethylaminopyridine and protoporphyrin is (2.0-2.5):(2.2-2.8):1; and / or, the condition parameters of the chelating reaction include: the cobalt source is anhydrous cobalt chloride, Co 2+ a molar ratio of the original porphyrin is (8-12) : 1, a reaction temperature is 25℃, and a reaction time is 12 h; And / or, the conditions and parameters of the amidation reaction include: using benzotriazole-N,N,N',N'-tetramethylurea hexafluorophosphate and N,N-diisopropylethylamine as a coupling system, wherein the molar ratio of benzotriazole-N,N,N',N'-tetramethylurea hexafluorophosphate, N,N-diisopropylethylamine and mono-(6-hexamethylenediamino-6-deoxy)-β-cyclodextrin is (1.2-1.5):(3.5-4.5):1, the reaction temperature is 25℃, and the reaction time is 48 h.

4. A modular polypeptide nanovaccine, characterized in that, include: carrier; The polypeptide and adjuvant loaded on the carrier; The carrier is a nanoparticle formed by the self-assembly of the compound of claim 1 or the compound prepared by any one of claims 2 to 3 in an aqueous phase. The polypeptide is selected from the TYRP-1 polypeptide with a 6His tag attached to the sequence shown in SEQ ID NO: 1 and the TATK polypeptide with a 6His tag attached to the sequence shown in SEQ ID NO:

2. The adjuvant is R848.

5. The modular polypeptide nanovaccine according to claim 4, characterized in that, The average particle size of the nanoparticles is 50-200 nm. And / or, the coefficient of variation of the dispersibility of the nanoparticles is ≤0.

3.

6. The modular polypeptide nanovaccine according to claim 4, characterized in that, The loading rate of the polypeptide is ≥95%.

7. The modular polypeptide nanovaccine according to claim 4, characterized in that, The adjuvant is loaded into nanoparticles via the hydrophobic cavity of mono-(6-hexamethylenediamino-6-deoxy)-β-cyclodextrin in the compound, and the loading rate of the adjuvant is ≥70%.

8. A method for preparing a modular polypeptide nanovaccine according to any one of claims 4 to 7, characterized in that, The preparation method of the modular polypeptide nanovaccine includes the following steps: The compound was dissolved in an organic solvent and then added dropwise to a PBS solution. The mixture was then stirred to evaporate the organic solvent and purified by a first filtration to obtain a nanoparticle solution. The nanoparticle solution was incubated with peptides and adjuvants, followed by a second filtration to obtain the modular peptide nanovaccine.

9. The method for preparing the modular polypeptide nanovaccine according to claim 8, characterized in that, The organic solvent includes at least one of tetrahydrofuran, methanol, and ethanol, and the concentration of the compound in the organic solvent is 2.0-3.0 mg / mL; the dropping rate is 45-55 μL / min, the stirring temperature is 37-40℃, and the stirring time is 12 h; the first filtration uses a 0.22 μm polyethersulfone membrane, and the second filtration uses an ultrafiltration tube with a molecular weight cutoff of 100 kDa; And / or, the mass ratio of the polypeptide to the nanoparticles is (0.1-0.2):1, and the mass ratio of the adjuvant to the nanoparticles is (0.05-0.1):1; And / or, the incubation conditions include: incubation at room temperature for 1.5 to 2.5 hours or incubation at 4°C for 10 to 14 hours.

10. The use of a modular polypeptide nanovaccine according to any one of claims 4 to 7, or a modular polypeptide nanovaccine prepared by the preparation method according to any one of claims 8 to 9, in the preparation of a drug for the prevention or treatment of melanoma.

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