PEGylated nano-particles based on pyridyl porphyrin as well as preparation method and application of PEGylated nano-particles

Abstract

The invention relates to a pyridylporphyrin-based pegylated nano-particle as well as a preparation method and application of the pyridylporphyrin-based pegylated nano-particle. The nano-particle is a spherical nano-particle with a shell-core structure; the shell is of a lipid-polyethylene glycol double-layer structure; the core is a platinum ion type compound based on pyridyl porphyrin; the platinum ion type compound based on the pyridyl porphyrin is prepared by mixing a pyridinol ligand and PtCl2 (PhCN) 2. The platinum drug and the protonated porphyrin are combined through ion pairs, so that many problems of the traditional platinum drug are solved, the water solubility of the platinum drug and the cytotoxic effect on tumor cells are mainly improved, and the synergistic anticancer effect of a chemical therapy and a PDT therapy is achieved. The material does not need a complex preparation process, is simple to operate and is easy to package.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical technology, and particularly to a polyethylene glycolated nanoparticle based on pyridyl porphyrin, a preparation method thereof, and an application thereof. Background Art

[0002] Skin cancer is a general term for skin malignancies, mainly divided into non-melanoma skin cancer and melanoma skin cancer. The former is further divided into two categories: basal cell carcinoma and squamous cell carcinoma, and it is prone to occur in areas such as the cheeks, nose, and eyelid skin that are exposed to sunlight for a long time. Ultraviolet radiation is the main cause of skin cancer. Ultraviolet irradiation can cause DNA damage and gene mutations, thereby inducing skin cancer. The incidence of skin cancer varies greatly in different regions.

[0003] Compared with non-melanoma, melanoma is a type of malignant tumor with high recurrence, high metastasis, and poor prognosis, and its incidence has been continuously increasing globally. Although melanoma is not as common as other skin cancers, it is more lethal. Melanoma is one of the most aggressive cancers and is more likely to metastasize. Melanoma can spread to other organs such as the liver, lungs, and brain through lymphatic and hematogenous metastasis in the early stage, and brain metastasis occurs in the terminal stage. The disease progresses rapidly in the brain metastasis stage, which is the main cause of death for patients.

[0004] Treatment options for melanoma include traditional surgical resection, conventional chemotherapy, emerging targeted therapy, and immunotherapy, etc. Photodynamic Therapy (PDT) is a new mode of treating malignant tumors that has emerged in recent years and has now become an emerging method for cancer treatment. PDT treatment can be traced back to ancient times. In 1900, German doctor Oscar Rabb first reported the chemical sensitization effect of light on tissues. In 1903, Jesionek and Tappeiner treated skin cancer patients with eosin and sunlight, marking the beginning of PDT therapy. In the 1960s, Lipson and Schwartz discovered HpD (hematoporphyrin derivative), and in the early 1980s, they isolated and identified the active components of hematoporphyrin derivative. Subsequently, scientists found that the "active" part of HpD consists of porphyrin dimers and oligomers, and the porphyrin units can be connected through ethers, esters, and carbon-carbon bonds. PDT generates reactive oxygen species (ROS) through the reaction of excited photosensitizers and oxygen sources, such as singlet oxygen ( 1 O2), hydroxyl radical (·OH), and superoxide radical (·O2 -) to kill tumor cells. Although most PDT processes are related to oxygen and initiated under oxygen-containing conditions, there are also some PDTs that occur under hypoxic environments. According to different photoreactions, PDT has two types, namely type I and type II. After photoactivation, the photosensitizer first transitions from the ground singlet state (S0) to the excited singlet state (S1), and then to the electronically excited triplet state (T1); T1 then triggers photoreactions through two different pathways. For type I PDT, T1 participates in hydrogen or electron transfer processes, directly forming free radicals with biological substrates, and this free radical can further interact with triplet oxygen ( 3 O2) and water to generate ·O2 - and ·OH. In type II PDT processes, T1 undergoes type II photoreactions, directly converting the surrounding 3 O2 into cytotoxic 1 O2. Currently, most of the photosensitizers used mainly undergo type II photoreactions, and it is generally believed that 1 O2 is the main destroyer of photodynamic action on cancer cells.

[0005] Selective uptake of the photosensitizer by local lesions and exposure to light of an appropriate wavelength, through photosensitizer mediation and the participation of oxygen molecules, lead to oxidative damage, causing apoptosis and necrosis of target cells. Because it has almost no side effects on normal tissue cells, it has unique advantages clinically and has been widely used in the treatment of tumors such as esophageal cancer, non-melanoma skin cancer, and cervical cancer, and is expected to become a routine adjuvant treatment method for various malignant tumors. At the same time, PDT treatment can also induce the occurrence of tumor immunity, providing new strategies for the effective treatment of some refractory malignant tumors. Currently, new molecules with ideal properties have been found to be used as photosensitizers in PDT treatment. Among these characteristics, enhanced photostability, good solubility in physiological media, generation of high levels of reactive oxygen species (ROS), selectivity, and high phototoxicity are particularly emphasized. There is a rich variety of photosensitizers available for photodynamic therapy. Among them, porphyrins, as commonly used photosensitizers, have excellent photophysical properties due to their structure being a cyclic structure with 18 conjugated π electrons, such as Photofrin, Temoporfin, Photochlor, Redaporfin, etc. based on the porphyrin skeleton, and some of them have been approved by the FDA for clinical cancer treatment. To strengthen the role of these structures, scientists have studied the combination of porphyrins with some inorganic compounds, such as platinum complexes.

[0006] Platinum-based drugs such as cisplatin, carboplatin, and oxaliplatin have been widely used in the treatment of bladder cancer, ovarian cancer, head and neck cancer, testicular cancer, and lung cancer. However, despite these drugs being first-line drugs for clinical use, they all have serious side effects when used, such as nephrotoxicity, ototoxicity, hepatotoxicity, gastrointestinal dysfunction, and cardiotoxicity, etc. In addition, many tumors have intrinsic or acquired resistance to platinum-based drugs, which all limit the clinical use of platinum-based drugs. In recent years, researchers have put a lot of effort into developing new platinum-based anticancer complexes, but so far only a very small number of platinum-based anticancer complexes have been clinically applied worldwide. For example, Lipoplatin is a nano-drug formed by encapsulating cisplatin in liposomes with a size of about 110 nm.

[0007] The coordination of porphyrin with platinum complexes can provide an important way to generate compounds with both cytotoxicity and PDT characteristics to achieve a synergistic therapeutic effect and reduce the side effects caused by each drug used alone. Currently, many platinum complexes that can be activated by infrared light have been developed. These complexes all exhibit excellent singlet oxygen quantum yields, indicating that they all have the potential to be used as PDT drugs. In addition, the presence of heavy metal platinum increases the number of triplets in the photoexcited state and promotes the photochemical degradation pathway during the photophysical decay process. Compared with divalent platinum preparation Pt (II) Compared with, Pt (IV) -based prodrug complexes show negligible toxic side effects in vivo because they only release active platinum(II) analogs into the tumor environment, thus minimizing the interaction with blood proteins or other biomolecules.

[0008] In 1975, scientists first reported using HpD (hematoporphyrin derivative) to inhibit mammary fat tumors in mice. After that, porphyrins and their derivatives have been successively used in photodynamic therapy and are considered the first-generation photosensitizers. Among these porphyrin derivatives, the most representative one is Photofrin, which has been approved for clinical use. However, these porphyrin-based photosensitizers have poor selectivity for tumors and will accumulate highly in the skin, resulting in skin photosensitivity.

[0009] The second-generation photosensitizers include 5-aminolevulinic acid (5-ALA), phthalocyanine, chlorin e6, benzoporphyrin derivatives, bacteriochlorin analogs, etc. They have high selectivity for tumors and less accumulation in the skin. The maximum absorption of these photosensitizers is usually in the range of 630 - 800 nm, thus expanding the accessible depth of PDT. However, these hydrophobic photosensitizers have poor circulating half-lives and serious self-aggregation effects, both of which affect the overall therapeutic effect.

[0010] With the booming development of nanomedicine, materials scientists have developed a variety of nanocarriers for photosensitizer delivery, which can enhance the accumulation of photosensitizers in tumors. In 2006, the Kono group synthesized a polyethylene glycolylated dendritic material for loading protoporphyrin IX (PpIX) or hydrophobic rose bengal (RB) for PDT. In 2015, the Zhao Yanli group utilized the large cavity and high surface area of hollow mesoporous SiO2 nanoparticles (HMSNs) to synthesize folic acid-conjugated HMSNs for loading 5-ALA, thus achieving targeted PDT for B16F10 (melanoma). In 2009, the Wang Kemin group co-condensed the silanol groups between methylene blue (MB) and tetraethyl orthosilicate (TEOS) through a water-in-oil reverse microemulsion method to obtain MB-grafted SiO2 nanoparticles with almost no MB leakage during in vivo circulation and strong resistance to diaphorase. However, the long-term biosafety of these nanocarriers needs to be further evaluated before the third-generation photosensitizers are used clinically.

[0011] Platinum complexes are well-known for their antitumor activity. So far, thousands of platinum complexes have been prepared and evaluated as potential anticancer drugs. However, the tumor selectivity of platinum-based drugs is quite poor, and systemic toxicity and drug resistance have become the main drawbacks of platinum-based drugs. Therefore, chemical and biomedical researchers have shown great interest in functionalizing platinum complexes with bioactive molecules, photosensitizers, nanomaterials, etc. The motivation for functionalization comes from the following needs: improving tumor selectivity or minimizing the systemic toxicity of drugs, enhancing the cellular accumulation of drugs, overcoming tumor drug resistance, imaging drug molecules in vitro or in vivo, achieving synergistic anticancer effects between different treatment modalities, or adding additional functions to drugs.

[0012] In recent years, scientists have been trying to chemically bond platinum-based drugs to porphyrin derivatives to obtain synergistic PDT and chemotherapy anticancer activities from a single prodrug. For example, the Spingler group reported the synthesis of platinum-based porphyrin compounds from commercially available 5,10,15,20-tetrakis(4-pyridyl)porphyrin and differently substituted platinum complexes and studied their photophysical characteristics and in vitro photoinduced anticancer properties (Angew. Chem. 2014, 126, 7058 - 7061). The quantum yield of 1 O2 was determined by using a 9,10-dimethylanthracene-based photobleaching assay in N,N-dimethylformamide (DMF). The results showed that all the complexes exhibited excellent 1 O2 quantum yields, indicating that they all have the potential for PDT. In addition, these complexes are lipophilic cations. The chemical potential difference between the inner and outer layers of the cell membrane is favorable for the diffusion of cations, thus enhancing their cellular uptake (passive targeting).

[0013] Li et al. prepared a polymer nanoparticle by combining oxaliplatin and a polyethylene glycolated porphyrin photosensitizer (photosensitizer), achieving a synergistic anti-cancer effect of chemotherapy and PDT. The negative charge carried on the surface of the nanoparticle and polyethylene glycol (PEG) endow it with good stability in blood circulation, minimizing protein adsorption (Adv. Mater. 2019, 31, 1805888). When reaching the tumor site, the PEG on the particle surface is decomposed by the metalloproteinase MMP-2, and its surface charge changes from negative to positive in the acidic tumor microenvironment (pH = 6.8), thus enhancing the penetration of the nano-system into deep tumors and the drug delivery efficiency.

[0014] In 2019, the research group of Zhao designed a nano-drug (PEG-Por-CD:oxliPt(IV)-ada), which was synthesized by the host-guest interaction of an oxaliplatin prodrug (oxliPt(IV)-ada) and a porphyrin photosensitizer (PEG-Por-CD) through self-assembly (ACS Appl. Mater. Interfaces 2019, 11, 16391 - 16401). β-CD and ada were used to modify the porphyrin and the oxaliplatin prodrug respectively to prepare an amphiphilic host-guest complex, which was then synthesized by self-assembly. This nano-drug has good biocompatibility, can accumulate at the tumor site, and will decompose in a reducing environment to release the therapeutic drugs.

[0015] However, the existing platinum complexes have poor selectivity for tumors, and systemic toxicity and drug resistance are the main drawbacks. Although various nanoparticle-based platinum drug tumor-targeted drug delivery systems have been proposed, due to the clearance by the reticuloendothelial system (RES), the in vivo delivery of these nanostructures to the target site is still not optimal. In addition, the components of the supramolecular drug delivery system constructed by the research group of Zhao are extremely complex, and the feasibility of clinical use is relatively low. Therefore, using porphyrin and other functional molecules as building blocks to form compounds in the form of ion pairs and preparing multifunctional nanoparticles with controllable structures through simple nano-encapsulation technology can well solve the above problems. At the same time, the prepared nanoparticles have the EPR effect, which can increase the accumulation of nanoparticles at tumor cells. The self-assembled nanoparticles have good biocompatibility and a long blood circulation time, which are all beneficial to expanding their biological applications. Summary of the Invention

[0016] To solve the above technical problems, the present invention provides a pyridyl porphyrin-based polyethylene glycolated nanoparticle, a preparation method thereof, and an application thereof. The present invention synthesizes a platinum ionic compound [H8TMPP]2(PtCl6)5(Cl)2·2(HCl) based on pyridyl porphyrin that can stably exist under acidic conditions. Since its ultraviolet absorption spectrum has a broad and strong absorption peak in the range of 630-660 nm and is within the photodynamic therapy window, the present invention speculates that it has a high 1 O2 quantum yield and can be used as a photosensitizer in PDT. In addition, in order to improve the stability of the compound and extend the blood circulation time, a polyethylene glycolated nanoparticle, abbreviated as [H8TMPP]-Pt-PEG, is obtained by mixing a tetrahydrofuran solution of 1,2-dioleoyl-glycero-3-phosphocholine (DOPC), cholesterol, and distearoylphosphatidylethanolamine 2000 (DSPE-polyethylene glycol 2000) and an ethanol / water (v / v = 1 / 2) mixed solution of the ionic compound in a molar ratio of 2:2:1. The results of cell viability tests show that under the condition of no light irradiation, [H8TMPP]-Pt-PEG has a strong cytotoxic effect on mouse melanoma cells (B16F10), and the IC 50 value is 11.6 μg / mL. Under 650 nm laser irradiation, more than 90% of B16F10 cancer cells can be killed at a concentration of 0.45 μg / mL. At the same time, the anti-cancer experiment in mice also fully demonstrates the synergistic anti-cancer effect of chemotherapy and PDT therapy.

[0017] The present invention is realized through the following technical solutions:

[0018] The first object of the present invention is to provide a pyridyl porphyrin-based polyethylene glycolated nanoparticle, and the nanoparticle is a spherical nanoparticle with a core-shell structure; the shell is a lipid-polyethylene glycol bilayer structure; the core is a platinum ionic compound based on pyridyl porphyrin;

[0019] The platinum ionic compound based on pyridyl porphyrin is prepared by mixing a pyridino alcohol ligand and PtCl2(PhCN)2.

[0020] In an embodiment of the present invention, the lipid-polyethylene glycol bilayer structure is prepared from a lipid, a stabilizer, and distearoylphosphatidylethanolamine.

[0021] In an embodiment of the present invention, the lipid is selected from 1,2-dioleoyl-glycero-3-phosphocholine.

[0022] In an embodiment of the present invention, the stabilizer is selected from one or more of cholesterol, stigmasterol, and β-sitosterol.

[0023] In one embodiment of the present invention, the molecular weight of polyethylene glycol in the distearoylphosphatidylethanolamine is 500 Da - 5000 Da. Specifically, it includes distearoylphosphatidylethanolamine 2000, distearoylphosphatidylethanolamine 500, distearoylphosphatidylethanolamine 750, distearoylphosphatidylethanolamine 1000, distearoylphosphatidylethanolamine 5000, etc.; preferably distearoylphosphatidylethanolamine 2000.

[0024] In one embodiment of the present invention, the molar ratio of the lipid, the stabilizer, and the distearoylphosphatidylethanolamine is 2∶2∶1.

[0025] In one embodiment of the present invention, the pyridyl alcohol ligands are selected from one or more of meso-5,10,15,20-tetrakis(6-methyl-3-pyridyl)porphyrin, meso-5,10,15,20-tetrakis(6-ethyl-3-pyridyl)porphyrin, meso-5,10,15,20-tetrakis(6-isopropyl-3-pyridyl)porphyrin, meso-5,10,15,20-tetrakis(6-tert-butyl-3-pyridyl)porphyrin, and meso-5,10,15,20-tetrakis(6-phenyl-3-pyridyl)porphyrin.

[0026] The second object of the present invention is to provide a method for preparing the pyridyl porphyrin-based polyethylene glycolated nanoparticles, comprising the following steps:

[0027] (1) Mix the pyridyl alcohol ligand with PtCl2(PhCN)2 and acetic acid, add benzonitrile, and obtain a pyridyl porphyrin-based platinum ionic compound after heating;

[0028] (2) Mix the lipid and the stabilizer in an organic solvent, add an alcohol solution of the pyridyl porphyrin-based platinum ionic compound obtained in step (1) and distearoylphosphatidylethanolamine for a mixing reaction to obtain pyridyl porphyrin-based polyethylene glycolated nanoparticles.

[0029] The third object of the present invention is to provide the application of the pyridyl porphyrin-based polyethylene glycolated nanoparticles in the preparation of anti-tumor drugs.

[0030] In one embodiment of the present invention, the drug is a drug for combined photodynamic therapy and chemotherapy.

[0031] In one embodiment of the present invention, the tumor is selected from one or more of melanoma, glioma, colon cancer, lung cancer, breast cancer, liver cancer, and ovarian cancer.

[0032] The present invention utilizes a hydrothermal reaction of the H2TMPP ligand with PtCl2(PhCN)2 to obtain a highly water-soluble salt compound [H8TMPP]2(PtCl6)5(Cl)2·2(HCl). Since it has a strong absorption peak at 630 - 660 nm, within the range of the phototherapy window, it is speculated that this compound has a high 1 O2 quantum yield and can be applied to the combined chemotherapy and PDT treatment of tumors. To overcome the drawback that it can only exist stably under strong acid (pH < 1.9) conditions and expand its biological applications, a polyethylene glycolylation scheme is adopted. Using the hydrophilic-hydrophobic interaction, [H8TMPP]2(PtCl6)5(Cl)2·2(HCl) is encapsulated with cholesterol / distearoyl phosphatidylethanolamine-polyethylene glycol 2000 (DSPE-PEG2000) / dioleoyl phosphatidylcholine (DOPC) to prepare nanoparticles (labeled as [H8TMPP]-Pt-PEG).

[0033] The present invention uses porphyrins and their derivatives with different degrees of protonation as cations, and simultaneously uses metal ions and compounds with biological activities (including anticancer activities) as anions to form ion pair compounds. On this basis, biocompatible polymer materials are used to encapsulate them to prepare nanoparticle materials, and the synergistic treatment of tumors (such as melanoma, glioblastoma, colon cancer, lung cancer, breast cancer, liver cancer or ovarian cancer) by chemotherapy and PDT using the obtained materials is studied.

[0034] The above technical solutions of the present invention have the following advantages compared with the prior art:

[0035] (1) The present invention solves many problems of traditional platinum drugs by combining platinum drugs with protonated porphyrins through ion pairs, mainly including improving the water solubility of platinum drugs and their cytotoxic effects on tumor cells, and realizing the synergistic anticancer effect of chemotherapy and PDT therapies. The material does not require a complex preparation process, is easy to operate, and is easy to encapsulate.

[0036] (2) In the material of the present invention, platinum shows a tetravalent state (Pt IV) and is free outside the protonated porphyrin in the form of hexachloroplatinate ions. Utilizing the relatively easy interconversion between platinum oxidation states, reducing the non-cytotoxic octahedral platinum(IV) compound to the significantly cytotoxic square planar platinum(II) complex inside cells can avoid tumor cell hypoxia to sensitize PDT. In addition, by mixing a tetrahydrofuran solution of 1,2-dioleoyl-glycero-3-phosphocholine (DOPC), cholesterol, and distearoylphosphatidylethanolamine 2000 (DSPE-PEG2000) in a molar ratio of 2:2:1 and an ethanol / water (v / v = 1 / 2) mixed solution of the ionic compound, PEGylated nanoparticles are obtained, improving the stability of the compound and prolonging the blood circulation time.

[0037] (3) Compared the antitumor effects, in vivo distribution, and blood circulation time of cisplatin and [H8TMPP]-Pt-PEG in vitro / in vivo. Compared with cisplatin, [H8TMPP]-Pt-PEG nanoparticles show excellent combined chemo- and PDT anti-cancer efficacy at the cellular and animal levels due to advantages such as enhanced enhanced permeability and retention (EPR) effect, prolonged blood circulation time, and increased tumor accumulation. Brief Description of the Drawings

[0038] To make the content of the present invention easier to be clearly understood, the following further details the present invention according to specific embodiments of the present invention in combination with the drawings, where

[0039] Figure 1 is the synthesis schematic diagram and crystal structure diagram of [H8TMPP]2(PtCl6)5(Cl)2·2(HCl) in the present invention;

[0040] Figure 2 . is the synthesis schematic diagram of [H8TMPP]-Pt-PEG in the present invention;

[0041] Figure 3 is the PXRD pattern of [H8TMPP]2(PtCl6)5(Cl)2·2(HCl) in the present invention;

[0042] Figure 4 is the infrared spectrum of [H8TMPP]2(PtCl6)5(Cl)2·2(HCl) in the present invention;

[0043] Figure 5 is the UV-Vis spectrum of H2TMPP and [H8TMPP]2(PtCl6)5(Cl)2·2(HCl) in the present invention;

[0044] Figure 6It is the ultraviolet absorption spectrum of [H8TMPP]2(PtCl6)5(Cl)2·2(HCl) in aqueous solutions with different pH values in the present invention;

[0045] Figure 7 It is the X-ray photoelectron spectroscopy (X photosensitizer) characterization of [H8TMPP]2(PtCl6)5(Cl)2·2(HCl) in the present invention;

[0046] Figure 8 It is the TEM image and hydrodynamic diameter diagram of [H8TMPP]-Pt-PEG in the present invention;

[0047] Figure 9 It is the cell survival rate diagram (a) and the corresponding half-inhibitory concentration curve (b) of B16F10 cells after being treated with different treatment groups for 12 h in the present invention;

[0048] Figure 10 It is the fluorescence microscope image of B16F10 cells after incubation with [H8TMPP]-Pt-PEG in the present invention; The cell nuclei were stained with Hoechst 33258;

[0049] Figure 11 It is the fluorescence microscope image of B16F10 cells treated with [H8TMPP]-Pt-PEG and DCFH-DA and irradiated with laser (650 nm, 25 mW cm -2 ) in the present invention;

[0050] Figure 12 It is the flow cytometry diagram of (a) cell uptake and (b) intracellular ROS of B16F10 after incubation with [H8TMPP]-Pt-PEG for 2 h in the present invention;

[0051] Figure 13 It is the relationship diagram between the concentration of [H8TMPP]-Pt-PEG and the hemolysis rate in the present invention;

[0052] Figure 14 It is the in vivo anti-tumor effect in the present invention. (a) Tumor growth inhibition curve; (b) Mouse weight change curve; (c) Survival rate curve of mice in each treatment group; (d) Tumor weight diagram of each treatment group; (e) Tumor photos of each treatment group;

[0053] Figure 15 It is the tissue distribution of (a) [H8TMPP]-Pt-PEG and (b) cisplatin after intravenous injection of the material in the present invention. Organs: heart (H), lung (Lu), liver (Li), kidney (K), spleen (Sp), tumor (T), (c) Ex vivo fluorescence imaging diagrams of mice 2 h (left figure) and 8 h (right figure) after porphyrin injection;

[0054] Figure 16 It is the time-dependent curve of platinum concentration in plasma after intravenous injection of cisplatin and [H8TMPP]-Pt-PEG nanoparticles in the present invention. Detailed implementation manners

[0055] The present invention will be further described below in conjunction with the accompanying drawings and specific embodiments, so that those skilled in the art can better understand the present invention and be able to implement it, but the specific embodiments cited are not intended to limit the present invention.

[0056] The present invention synthesized a platinum-based ionic compound [H8TMPP]2(PtCl6)5(Cl)2·2(HCl) that can stably exist under acidic conditions, and determined its structure and composition through X-ray single crystal diffraction, elemental analysis, X-ray powder diffraction, Fourier transform infrared, energy dispersive spectrometer, ultraviolet-visible spectrophotometer, etc. Since its ultraviolet absorption spectrum has a strong broad absorption peak in the range of 630-660 nm, within the range of the PDT treatment window, it is speculated that it has a high 1 O2 quantum yield and can be applied to the combined treatment of tumor chemotherapy and PDT. In subsequent studies, the present invention used DOPC / cholesterol / DSPE-PEG 2000 as raw materials to carry out polyethylene glycolylation modification on it, and obtained a water-soluble polyethylene glycolylated product [H8TMPP]-Pt-PEG that can stably exist under neutral conditions. In a series of in vitro and in vivo experiments, the present invention found that compared with the common anticancer drug cisplatin, due to the enhanced EPR effect, [H8TMPP]-Pt-PEG has a longer blood circulation time and increased tumor accumulation. After combining with laser treatment at 650 nm, the growth of tumors was completely inhibited, fully demonstrating the combined anticancer effect of chemotherapy and photodynamic therapy of [H8TMPP]-Pt-PEG. The materials used in the present invention are simple to prepare, have a high yield, and have a good synergistic effect between PDT and chemotherapy, and are a class of antitumor materials with potential application value.

[0057] Unless otherwise specified, the experimental methods used in the following examples are all conventional methods, and the materials, reagents, etc. used, unless otherwise specified, can be obtained from commercial channels.

[0058] B16F10 cells (mouse melanoma cells) were obtained from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China). The culture medium was DMEM supplemented with 10% FBS and 1% penicillin / streptomycin, and the cells were cultured in a cell incubator at 37°C and 5% CO2. Cells in the logarithmic growth phase were taken for experiments.

[0059] BALB / c nude mice, female, 6 - 8 weeks old, weighing 18 - 22 g, were purchased from Zhejiang Academy of Medical Sciences. The animals were housed in a constant temperature environment of 18 - 22°C and a constant humidity environment of 50 - 80% in a closed and sterile environment. Animal experiments were conducted in accordance with the "China Animal Protection Law" (CAPN). All animal experiments complied with the relevant regulations of the Laboratory Animal Management Committee.

[0060] Statistical analysis of data: Experimental data were expressed as mean ± standard deviation. Analysis of variance (ANOVA) and Student's t - test were used to determine significant differences between groups ( * P < 0.05, ** P < 0.01, *** P < 0.001 and **** P < 0.0001).

[0061] Example 1 Synthesis and Physicochemical Characterization of [H8TMPP] - Pt - PEG Material

[0062] 1. Synthesis of Compound PtCl2(PhCN)2

[0063] Add K2PtCl4 (0.100 g, 0.241 mmol) and 10 mL of distilled water to a round - bottom flask and stir to obtain a clear solution. Add benzonitrile (0.120 g, 1.163 mmol) to the above - mentioned solution and stir the mixture at room temperature for three days until the initial solution changes from red to colorless. After stirring, a green solid precipitates, and then it is separated by extraction with CH2Cl2. The extract is dried over anhydrous sodium sulfate and then filtered. The liquid is removed by evaporation under reduced pressure to obtain a bright - green solid crude product. The crude product is washed with diethyl ether 2 - 3 times and dried under vacuum to obtain pure PtCl2(PhCN)2 (0.097 g, yield 85%).

[0064] 1 1H NMR (400 MHz, CDCl3): δ 7.88 - 7.78 (m, 4H), 7.78 - 7.71 (m, 2H), 7.61 - 7.52 (m, 4H).

[0065] Synthesis of Compound [H8TMPP]2(PtCl6)5(Cl)2·2(HCl) (The synthesis schematic diagram is as Figure 1 shown; The preparation method of H2TMPP refers to Patent CN107903272B)

[0066] meso-5,10,15,20-Tetrakis(6-methyl-3-pyridyl)porphyrin (H2TMPP, 1.0 mg, 0.0015 mmol), PtCl2(PhCN)2 (2.0 mg, 0.0042 mmol) and acetic acid (20 μL) were placed in a Pyrex glass tube, dissolved in benzonitrile and transferred to a programmable oven. The mixture was heated to 120 °C in 3 h and maintained at this temperature for 48 h, then cooled to room temperature in 12 h to obtain purple-black rod-shaped crystals. The solid product was collected by centrifugation, washed three times with ether, and dried under vacuum to obtain the solid product of [H8TMPP]2(PtCl6)5(Cl)2·2(HCl).

[0067] 3. Synthesis of [H8TMPP]-Pt-PEG (the synthesis schematic diagram is as Figure 2 shown)

[0068] DOPC (30.1 mg, 0.038 mmol) and cholesterol (13.0 mg, 0.034 mmol) were mixed in 20 mL of THF, stirred evenly and added to a 50 mL round-bottom flask, and stirred under reflux. Separately, [H8TMPP]2(PtCl6)5(Cl)2·2(HCl) (30.8 mg, 0.0176 mmol) was added to a mixed solvent of H2O / EtOH (20 mL / 10 mL). At first, the dissolution was slow, but the solid dissolved immediately after adding a few drops of concentrated HCl. The [H8TMPP]2(PtCl6)5(Cl)2·2(HCl) solution was added to the above THF solution. After refluxing for 1 h, DSPE-PEG 2000 (16.3 mg, 0.00815 mmol) was added to the above mixed solution. After refluxing overnight at 80 °C, it was dialyzed for 48 h with a dialysis bag with a molecular weight cut-off of 3500 to obtain an aqueous solution of [H8TMPP]-Pt-PEG, which was stored in a -80 °C refrigerator for later use.

[0069] 4. Characterization of materials

[0070] The above-obtained platinum porphyrin ionic compound [H8TMPP]2(PtCl6)5(Cl)2·2(HCl) and the polyethylene glycolated [H8TMPP]-Pt-PEG were respectively characterized by X-ray single crystal diffraction (SCXRD), elemental analysis, X-ray powder diffraction (PXRD), Fourier transform infrared (FT-IR), ultraviolet-visible absorption (UV-Vis) and transmission electron microscopy (TEM) etc. to determine their structural characteristics. The specific characterizations are summarized as follows:

[0071] (1) The X-ray single crystal diffraction (SCXRD) data of [H8TMPP]2(PtCl6)5(Cl)2·2(HCl) are as follows: Molecular formula C 88 H 80 Cl 33 N 16 Pt5; Molecular weight 3506.98; Crystal system orthorhombic; Space group Pbam; 15.0532(8); 30.0670(17); 13.9586(8); α / °90; β / °90; γ / °90; 6317.7(6); Z 2; D c / (g cm -3 ) 1.844; μ(Mo-Kα) / mm -1 6.260; F(000) 3342; Total number of diffraction points 177306; Number of independent diffraction points 7894; Number of observable diffraction points (I>2σ(I)) 4178; R index [I>2σ(I)] R1 = 0.0953, wR2 = 0.2491; GOF 1.136.

[0072] (2) Structure description of [H8TMPP]2(PtCl6)5(Cl)2·2(HCl).

[0073] The ionic platinum-based porphyrin compound [H8TMPP]2(PtCl6)5(Cl)2·2(HCl) in the form of purple-black rods was obtained by hydrothermal method, and its structure was determined by an X-ray single crystal diffractometer. From the single crystal structure ( Figure 2 ), it can be seen that the four pyridine Ns in the H2TMPP ligand and the two pyrrole Ns at the center of the porphyrin skeleton are completely protonated, forming a +6 valence cationic structure. In this structure, the porphyrin skeleton is deformed due to the lack of conjugation. [PtCl6] 2- ions are free outside the protonated porphyrin ring, thus maintaining the valence balance of the whole ionic compound.

[0074] (3) Powder diffraction pattern of [H8TMPP]2(PtCl6)5(Cl)2·2(HCl):

[0075] As Figure 3 shown, the actual powder diffraction pattern of the crystal obtained from the reaction of H2TMPP and PtCl2(PhCN)2 is basically consistent with the powder diffraction pattern generated by fitting the single crystal structure, proving that the structure of the obtained crystal powder is consistent with the single crystal structure and is [H8TMPP]2(PtCl6)5(Cl)2·2(HCl).

[0076] (4)Elemental analysis (%) of [H8TMPP]2(PtCl6)5(Cl)2·2(HCl): C 88 H 80 Cl 33 N 16 For Pt5: Theoretical values: C 30.11, H 2.28, N 6.39; Measured values: C 29.74, H 2.75, N 6.60.

[0077] (5)Infrared spectrum of [H8TMPP]2(PtCl6)5(Cl)2·2(HCl) Figure 4 ) and data: 3440.85(m), 3098.53(m), 3031.48(m), 2906.38(w), 2839.29(w), 2579.16(w), 2162.94(w), 2046.08(w), 1638.56(s), 1601.99(vs), 1545.78(s), 1494.69(s), 1450.56(m), 1380.49(w), 1288.66(s), 1234.5(s), 1142.64(s), 1084.21(m), 1043.8(vs), 984.39(vs), 882.46(s), 853.73(s), 823.98(vs), 801.71(vs), 714.17(vs), 643.34(s).

[0078] (6)Ultraviolet absorption spectrum of [H8TMPP]2(PtCl6)5(Cl)2·2(HCl):

[0079] Figure 5 are the ultraviolet absorption spectra of H2TMPP and [H8TMPP]2(PtCl6)5(Cl)2·2(HCl). It can be found that the ultraviolet absorption peak of the protonated platinum porphyrin ionic compound undergoes a red shift, and the Q band decreases. Moreover, a strong absorption peak appears at a wavelength of 635 nm, within the corresponding PDT treatment window range. It is speculated that this compound has the potential for photodynamic therapy.

[0080] During the experiment, it was found that the color of the solution of this compound dissolved in aqueous solutions with different pH values was different. Therefore, the ultraviolet absorption spectra of the solutions were measured to determine its stability conditions. From Figure 6 it can be seen that this compound can only stably exist under strong acidic conditions with a pH not greater than 1.91, and the ultraviolet absorption spectrum remains unchanged.

[0081] (7)X-ray photoelectron spectrum of [H8TMPP]2(PtCl6)5(Cl)2·2(HCl).

[0082] To determine the valence state of platinum in [H8TMPP]2(PtCl6)5(Cl)2·2(HCl), X-ray photoelectron spectroscopy (X photosensitizer) tests were conducted in this invention. Figure 7 a is the Pt 4f spectrogram of the compound [H8TMPP]2(PtCl6)5(Cl)2·2(HCl). By comparing with the X photosensitizer spectrograms of platinum complexes in the literature, it was found that 72.6 eV and 75.9 eV respectively correspond to the 4f 7 / 2 and 4f 5 / 2 of Pt(II), and 74.2 eV and 77.5 eV respectively correspond to the 4f 7 / 2 and 4f 5 / 2 of Pt(IV). Figure 7 b is the Cl 2p spectrogram of the compound [H8TMPP]2([PtCl6])5(Cl)2. Among them, 198.0 eV corresponds to the Pt(IV)-Cl bond, and 199.5 eV corresponds to the Pt(II)-Cl bond. From the comprehensive analysis, Pt(II) and Pt(IV) coexist in [H8TMPP]2(PtCl6)5(Cl)2·2(HCl).

[0083] (8) TEM analysis of PEGylated [H8TMPP]-Pt-PEG.

[0084] It was encapsulated with cholesterol / DOPC / DSPE-PEG2000, and through non-covalent interactions, PEGylated [H8TMPP]-Pt-PEG nanoparticles were prepared. This scheme utilizes the hydrophobic / hydrophobic interactions between DOPC / cholesterol / DSPE-PEG2000 to form a self-assembled asymmetric lipid-PEG bilayer on the surface of the platinum ionic compound, where cholesterol acts as a stabilizer. Figure 8 is the TEM image measured by dropping the prepared aqueous solution of [H8TMPP]-Pt-PEG onto a copper grid. It can be seen that the PEGylated material forms spherical nanoparticles with a core-shell structure and a size of about 200 nm. And it was found that the hydrodynamic diameter measured by DLS is 10 - 20 nm larger than the particle size measured by TEM ( Figure 8 b inset), which can be attributed to the lipid layer of DOPC / cholesterol / DSPE-PEG2000 on the particle surface and the swelling of the particle core.

[0085] Example 2 Cytological experiments of [H8TMPP]-Pt-PEG

[0086] 1. Cytotoxicity experiment

[0087] The MTT method was used to analyze and evaluate the cytotoxicity of the material against B16F10 cells. Two 96-well plates were prepared for the phototoxicity and dark toxicity experiments of the material. B16F10 cells were inoculated at a density of 1.0×10 4 cells / well and cultured for 16 h at 37 °C under 5% CO2. For the phototoxicity experiment, the cells were co-incubated with a serum-free DMEM solution of [H8TMPP]-Pt-PEG containing gradient platinum ion concentrations (0.5 μg / mL, 1.0 μg / mL, 2.0 μg / mL, 4.0 μg / mL, 8.0 μg / mL) for 6 h, and then irradiated with light at a wavelength of 650 nm (25 mW cm -2 , 2 min), and then the cells were further cultured for 6 h. For the dark toxicity experiment, the cells were directly co-incubated with cisplatin and [H8TMPP]-Pt-PEG for 12 h. Then, MTT solution (200 μL, 0.5 mg mL -1 ) was added to each well, and the cells were cultured at 37 °C. After 4 h, the MTT solution was discarded by inverting the plate. DMSO (100 μL) was added to dissolve the formazan crystals of the metabolites, and the mixture was shaken on a shaker for 5 minutes, and then the absorbance at 570 nm was measured with a microplate reader. Cells without any treatment were used as controls.

[0088] To study the synergistic anti-cancer efficacy of [H8TMPP]-Pt-PEG, cytotoxicity experiments and PDT experiments were carried out in this invention. First, 2 mL of the original solution after dialysis described in the previous synthesis step was taken into a centrifuge tube, and 2 mL of 0.1 M PBS sample solution was added to obtain 4 mL of solution. 1 mL of the solution was taken, 2 mL of DMEM was added, and five sample solutions with concentration gradients were prepared by stepwise dilution.

[0089] Approximately 2 million B16F10 cells were suspended in DMEM medium containing 10% FBS (fatal bovine serun, fetal bovine serum) and 1% photosensitizer (Penicillin-streptomycin, penicillin-streptomycin; against), transferred to a culture dish, 10 mL of DMEM (10% FBS + 1% photosensitizer) was added, and the cells were cultured in an incubator at 37.5 °C under 5% CO2 for two days. Then the culture medium was removed. The cells were rinsed with 0.5 mL of trypsin (containing 0.25% EDTA), and then 2 mL of trypsin (containing 0.25% EDTA) was added and digested in the incubator for 3 minutes. The trypsin was neutralized with 3 mL of DMEM culture medium, and then centrifuged at 1000 rpm for 3 minutes. The solution was removed, and the cells were resuspended and centrifuged with 3 mL of DMEM, and the DMEM was removed.

[0090] After uniformly mixing the obtained cells with DMEM (6 mL) again, 10 μL was taken for counting, and it was found that there were approximately 1 million cells per milliliter. 1 mL of the cells was diluted with 12 mL of DMEM (10% FBS + 1% photosensitizer), and after pipetting evenly, they were plated in a 96-well MTT plate. The cells were added to the 96-well MTT plate, 200 μL per well, B1-F12. After incubating for 16 h, the culture medium was removed, different concentrations of drugs were added, and the PDT group was incubated for another 12 h and then irradiated with PDT light (650 nm, 100 mW; 6 min) and then placed in an incubator for 9 h. The non-PDT group was incubated for 21 h. The drugs were removed from both groups, 100 μL of MTT solution was added to each well and then incubated for another 4 h. The MTT was removed, and the resulting formazan was dissolved with 100 μL of DMSO per well. After shaking on a shaker for 3 min, the absorbance at 570 nm was measured with an enzyme-linked immunosorbent assay (ELISA) reader to obtain the corresponding survival rate. The relationship between the drug concentration and cell viability is as Figure 9 shown.

[0091] Figure 9 Cisplatin, the cytotoxicity results of [H8TMPP]-Pt-PEG, and [H8TMPP]-Pt-PEG + PDT. The experimental results showed that without laser irradiation, the cytotoxicity of cisplatin alone or [H8TMPP]-Pt-PEG (IC 50 value was 11.6 μg / mL) against B16F10 cells was relatively small. However, compared with cisplatin, the antitumor effect of [H8TMPP]-Pt-PEG was better, which might be due to the longer blood circulation time of the PEGylated porphyrin. When [H8TMPP]-Pt-PEG was combined with laser (650 nm, 25 mW cm -2 , 2 min) treatment, the killing effect on cancer cells was significantly enhanced ( Figure 9 a), and its IC 50 value was only about 2 μg / mL, approximately 1 / 18 of the IC 50 value of the cisplatin group. When the concentration of [H8TMPP]-Pt-PEG was 0.45 μg / mL, more than 90% of B16F10 cells could be killed, fully demonstrating the synergistic anti-cancer effect of chemotherapy and photodynamic therapy ( Figure 9 b).

[0092] 2. Cellular uptake experiment

[0093] After incubating B16F10 cells with [H8TMPP]-Pt-PEG with a porphyrin concentration of 30 μg / mL, the uptake of [H8TMPP]-Pt-PEG by the cells was observed under an inverted fluorescence microscope. The results are as Figure 10 shown. Red fluorescence of porphyrin appeared in the cells, and through blue and red co-localization, it could be seen that [H8TMPP]-Pt-PEG was successfully taken up into the cytoplasm. Figure 12Flow cytometry results of a also further demonstrated that [H8TMPP]-Pt-PEG was successfully taken up by cells.

[0094] 3. Detection of intracellular ROS content

[0095] The generation of intracellular ROS in B16F10 cells under light irradiation was measured using 2,7-dichlorodihydrofluorescein diacetate (DCFH-DA). As Figure 11 can be seen, compared with the control group, green fluorescence of DCF appeared in the cells treated with [H8TMPP]-Pt-PEG, indicating the generation of ROS in the cells, which was also further demonstrated in the Figure 12 flow cytometry results of b.

[0096] 4. Hemolysis experiment

[0097] Blood was collected from mice by enucleation of the eyeballs, and the blood compatibility of [H8TMPP]-Pt-PEG was evaluated by the hemolysis experiment. After co-incubation with red blood cells at 37 °C for 4 h, the hemolysis rate of [H8TMPP]-Pt-PEG within the concentration range of 0.025 - 0.5 mg / mL was less than 2% ( Figure 13 ). It shows that [H8TMPP]-Pt-PEG has good biocompatibility and is suitable for biological applications.

[0098] Example 3 Zoological experiments of [H8TMPP]-Pt-PEG

[0099] 1. Establishment of animal tumor models

[0100] Under sterile conditions, a cell suspension of B16F10 (100 μL, 2×10 6 cells) was subcutaneously injected into the right flank region of BALB / c nude mice. When the average tumor volume reached 90 mm 3 , grouping experiments were carried out, and this day was set as day 0.

[0101] 2. Evaluation of in vivo antitumor performance of photochemotherapy using BALB / C tumor-bearing nude mice

[0102] Nude mice with similar tumor sizes were selected and randomly divided into four groups: PBS, Cisplatin, [H8TMPP]-Pt-PEG, [H8TMPP]-Pt-PEG + L, with 5 mice in each group. Mice received PBS, Cisplatin (3.0 mg Pt / kg), and [H8TMPP]-Pt-PEG (3.00 mg Pt / kg) by tail vein injection, twice a week. For the [H8TMPP]-Pt-PEG + L group, laser treatment was performed for 8 minutes (650 nm, 100 mW cm -2, irradiated for 4 minutes every 10 seconds), and both drug administration and laser treatment were carried out for 3 weeks. During the treatment, the body weight of the mice was weighed every three days, and the tumor size was measured. The calculation formula for tumor volume is: (length) × (width) 2 / 2.

[0103] As Figure 14 shown, this invention recorded relevant data such as the tumor volume and the body weight of the mice during different treatments in each group. Figure 14 Figure b shows that there was no obvious change in the body weight of the mice in each group before and after the treatment, indicating that the intervention treatment did not affect the normal physiological metabolism of the mice. From Figure 14 the tumor growth inhibition curve in Figure a and Figure 14 the tumor weight graph in Figure d, it can be seen that compared with the control group injected with PBS, the tumor growth of the mice receiving single chemotherapy (cisplatin and [H8TMPP]-Pt-PEG) was inhibited to a certain extent, and due to the EPR effect of the nanoparticles, the drug uptake by cancer cells increased, resulting in a slightly better inhibitory effect of [H8TMPP]-Pt-PEG nanoparticles on tumors and a higher survival rate of the mice ( Figure 14 Figure c). From Figure 14 Figures a, d and e in it can be seen that after [H8TMPP]-Pt-PEG was combined with laser treatment, the growth of tumors could be completely inhibited, and the survival rate of the mice during the treatment was 100%. However, due to the limited penetration of the laser and the fact that the penetrated cancer cells were far from the epidermis, it was almost impossible to completely remove the primary tumor only by single photodynamic therapy.

[0104] 3. In vivo drug distribution study

[0105] Taking advantage of the property that porphyrin has red emission, nude mice injected with [H8TMPP]-Pt-PEG (3.00 mg Pt / kg, n = 5) via the tail vein were sacrificed after 2 h and 8 h, and the heart, liver, spleen, lungs, kidneys and tumors were taken out. The distribution pictures of the material in each organ and tumor were observed and photographed through a small animal in vivo fluorescence imaging system (M-MSI-EX, CRI). Moreover, in order to quantitatively compare the distribution of the material in different organs at different times in vivo, the collected organs and tumors were ground, made into homogenates and then centrifuged. A certain amount of supernatant was taken and lysed with nitric acid, and the lysate was analyzed and measured by ICP-MS to determine the platinum ion content in each organ and tumor at 2 h and 8 h. At the same time, the mice in the cisplatin group (3 mg Pt / kg) were also treated under the same conditions to determine the distribution of cisplatin in each organ and tumor in vivo at 2 h and 8 h.

[0106] The biodistributions of cisplatin and [H8TMPP]-Pt-PEG in tumors and other organs are as Figure 15As shown. According to the results of ICP-MS, due to rapid clearance, the accumulation of cisplatin in tumors is insufficient, only 0.69 μg / g, while it accumulates more in the liver, lungs, kidneys and spleen, which will cause greater toxic side effects. In contrast, due to the prolonged circulation time of [H8TMPP]-Pt-PEG in vivo, the accumulation of porphyrin in tumors is quite remarkable, and the concentration increases with time. After 8 hours of injection of [H8TMPP]-Pt-PEG, the platinum concentration in the tumor is 2.55 μg / g, which is about 3.7 times higher than that of cisplatin. The prolonged blood circulation time and increased tumor accumulation result in better anti-tumor effects of [H8TMPP]-Pt-PEG.

[0107] 4. Pharmacokinetic study of [H8TMPP]-Pt-PEG

[0108] At different time points after mice received cisplatin (3 mg Pt / kg) or [H8TMPP]-Pt-PEG (3 mg Pt / kg) by tail vein injection, blood was collected by enucleation of the eyeballs, and ICP-MS was used to measure the concentration of platinum in plasma to evaluate the blood circulation time of cisplatin and [H8TMPP]-Pt-PEG. As Figure 16 shown, within the first 5 hours after cisplatin injection, the platinum concentration decreased rapidly, and by 12 hours, cisplatin had been basically completely cleared from the blood. In contrast, the platinum concentration in plasma decreased more slowly after injection of [H8TMPP]-Pt-PEG, and there was still a relatively high platinum concentration after 26 hours, and the blood circulation time was significantly prolonged. In addition, Figure 16 it also shows that the total area under the [H8TMPP]-Pt-PEG curve is larger than that of cisplatin, indicating that the circulation behavior of porphyrin has been improved, which is attributed to the bilayer encapsulation of porphyrin by lipids and polyethylene glycol to form stable nanoparticles.

[0109] Obviously, the above embodiments are only examples for clear illustration and are not limitations on the implementation manners. For those of ordinary skill in the art, other different forms of changes or modifications can be made based on the above description. It is not necessary and impossible to enumerate all the implementation manners here. And the obvious changes or modifications derived therefrom are still within the protection scope of the present invention.

Claims

1. A polyethylene glycolated nanoparticle based on pyridyl porphyrin, characterized in that, The nanoparticle is a spherical nanoparticle with a core-shell structure; the shell is a lipid-polyethylene glycol bilayer structure; the core is a platinum ion-based compound based on pyridyl porphyrin; The platinum ion-based compound based on pyridyl porphyrin is prepared by mixing a pyridyl alcohol ligand with PtCl2(PhCN)2.

2. The nanoparticle according to claim 1, wherein The lipid-polyethylene glycol bilayer structure is prepared from lipids, stabilizers, and distearoyl phosphatidylethanolamine.

3. The nanoparticle according to claim 2, characterized in that, The lipid is selected from 1,2-dioleoyl-glycerol-3-phosphocholine.

4. The nanoparticle according to claim 2, characterized in that, The stabilizer is selected from one or more of cholesterol, stigmasterol, and β-sitosterol.

5. The nanoparticle according to claim 2, wherein, The molecular weight of the polyethylene glycol in the distearoyl phosphatidylethanolamine is 500 Da - 5000 Da.

6. The nanoparticle according to claim 1, characterized in that, The pyridyl alcohol ligand is selected from one or more of meso-5,10,15,20-tetrakis(6-methyl-3-pyridyl)porphyrin, meso-5,10,15,20-tetrakis(6-ethyl-3-pyridyl)porphyrin, meso-5,10,15,20-tetrakis(6-isopropyl-3-pyridyl)porphyrin, meso-5,10,15,20-tetrakis(6-tert-butyl-3-pyridyl)porphyrin, and meso-5,10,15,20-tetrakis(6-phenyl-3-pyridyl)porphyrin.

7. The preparation method of the pyridyl porphyrin-based polyethylene glycolated nanoparticles according to any one of claims 1-6, characterized in that, It includes the following steps: (1) Mix a pyridyl alcohol ligand with PtCl2(PhCN)2 and acetic acid, add benzonitrile, and obtain a platinum ion-based compound based on pyridyl porphyrin after heating; (2) Mix lipids and stabilizers in an organic solvent, add an alcohol solution of the platinum ion-based compound based on pyridyl porphyrin obtained in step (1) and distearoyl phosphatidylethanolamine for a mixing reaction to obtain a polyethylene glycolylated nanoparticle based on pyridyl porphyrin.

8. Use of the polyethylene glycolylated nanoparticle based on pyridyl porphyrin according to any one of claims 1-6 in the preparation of a drug for treating tumors.

9. The application according to claim 8, characterized in that, The drug is a drug for combined photodynamic therapy and chemotherapy.

10. The application according to claim 8, wherein The tumor is selected from one or more of melanoma, glioma, colon cancer, lung cancer, breast cancer, liver cancer, and ovarian cancer.

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