阅读说明：本技术 一种聚卟啉类化合物的制备方法及应用 (Preparation method and application of polyporphyrin compound ) 是由 郑楠 张芷伊 谢丹 宋汪泽 郑玉斌 于 2019-11-06 设计创作，主要内容包括：本发明属于高分子材料、生物医药材料及光动力治疗技术领域,涉及一种聚卟啉类化合物的制备方法及应用。将四羧酸修饰的四苯基卟啉溶解于有机溶剂中,浓度为0.1M-1M；随后在反应体系中加入末端双氨基的缩硫酮共聚单体,最后加入1-乙基-(3-二甲基氨基丙基)碳酰二亚胺盐酸盐以及4-二甲氨基吡啶。将反应体系升温至50℃-100℃,反应24h-72h。反应结束后,冷却到室温后沉降离心,粗产物用去离子透析,冻干,得到聚卟啉类化合物pPS。(The invention belongs to the technical field of high polymer materials, biomedical materials and photodynamic therapy, and relates to a preparation method and application of a polyporphyrin compound. Dissolving tetraphenylporphyrin modified by tetracarboxylic acid in an organic solvent, wherein the concentration of the tetraphenylporphyrin is 0.1-1M; then the thioketal comonomer with terminal diamino group is added into the reaction system, and finally 1-ethyl- (3-dimethylaminopropyl) carbonyldiimine hydrochloride and 4-dimethylaminopyridine are added. Heating the reaction system to 50-100 ℃ and reacting for 24-72 h. After the reaction is finished, cooling to room temperature, settling and centrifuging, and performing deionized dialysis and freeze-drying on the crude product to obtain the polyporphyrin compound pPS.)

1. The preparation method of the polyporphyrin compound is characterized by comprising the following steps:

dissolving tetraphenylporphyrin modified by tetracarboxylic acid in an organic solvent, wherein the concentration of the tetraphenylporphyrin is 0.1-1M; adding a thioketal comonomer with terminal diamino, finally adding 1-ethyl- (3-dimethylaminopropyl) carbonyldiimine hydrochloride and 4-dimethylaminopyridine into the reaction system, wherein the molar ratio of the tetra-carboxylic acid modified tetraphenylporphyrin, the thioketal comonomer with terminal diamino, 1-ethyl- (3-dimethylaminopropyl) carbonyldiimine hydrochloride and 4-dimethylaminopyridine is 1: 2.5-1.6: 4.8-8: 0.8-8, wherein the ratio of carboxylic acid to amino in the comonomer is 0.8-1.2; heating the reaction system to 50-100 ℃, and reacting for 24-72 h; after the reaction is finished, cooling to room temperature, then carrying out sedimentation centrifugation for 3-10 times by using a sedimentation agent, putting the crude product in a dialysis bag with the molecular weight cutoff of 100-1000, dialyzing for 3-5 days by using deionized water, and freeze-drying to obtain the polyporphyrin compound pPS, wherein the reaction formula is as follows:

tetracarboxylic acid modified tetraphenylporphyrin terminal diamino thioketal comonomer pPS

The polyporphyrin compound pPS is characterized in that:

the number average molecular weight is 7000 to 12000;

zeta potential: 7.2mV to-3.5 mV;

the molecular weight distribution PDI is 1.36-1.87.

2. The method according to claim 1, wherein the organic solvent is N, N-dimethylformamide or tetrahydrofuran.

3. The method according to claim 1 or 2, wherein the settling agent is diethyl ether or n-hexane.

4. The polyporphyrin compound is used for photodynamic therapy of tumors.

Technical Field

The invention belongs to the technical field of high polymer materials, biomedical materials and photodynamic therapy, and relates to a preparation method and application of a polyporphyrin compound.

Background

Photodynamic therapy (PDT) is currently a revolutionary high-tech biotherapeutic method for the treatment of malignancies, and PDT has received much attention in the cancer treatment field over the past decade (adv. drug delivery. rev.2008,60, 1627-. Compared with the traditional three cancer treatment means such as operation, radiotherapy and chemotherapy, PDT well realizes the target selectivity and tissue specificity to tumor cells, and has the advantages of slight wound, small toxic and side effect, repeated illumination treatment and the like, thereby having great superiority.

The majority of photosensitizers which have been put into extensive experimental study and clinical application are porphyrins which are macromolecular heterocyclic compounds formed by bridging α -carbon atoms of four pyrrole rings through methine (═ CH-) and have a pi electron structure with a large conjugated system.hematoporphyrin derivatives (HPD) are the first photosensitizers approved for the market, but have weak absorption in the red region and cannot effectively penetrate target tissues, so that only primary and recurrent tumors on the surface layer can be locally destroyed.2015D has poor tissue selectivity, a large dose is required for achieving therapeutic effects, toxic and side effects are strong, skin photosensitization is easy to generate, and clinical therapeutic effects are affected (Biomaterials research 2018,22, 25-25; Anticancer AgentsMed.chem.2001, 2001, 194). the second photosensitizers overcome the disadvantages that the first photosensitizers have a short absorption wave length, tissue penetration, low singlet oxygen yield, and the like, the Tetraphenylporphyrin (TPP) is a second photosensitizers typically prepared in the second generation of photosensitizers, and has a strong photosensitizing effect of mutual reduction of the photosensitizing effect of the photosensitizers in the first photosensitizers in vivo, which are easily formed by the photosensitizers, and the photosensitizing nanoparticles of the photosensitizers are widely used as photosensitizers for the photosensitizing drugs with a strong photosensitizing effect of the photosensitizers of the photosensitizer in the first photosensitizers 2558-60, the photosensitizers are widely used for the photosensitizers, which are widely used for the photosensitizers, the.

Based on the background, the novel poly-photosensitizer is designed and invented, and the poly-porphyrin photosensitive drug with high molecular weight is prepared by taking degradable weak bonds which are flexible and respond to a tumor microenvironment as a comonomer and carrying out copolymerization research with tetracarboxylic acid tetraphenylporphyrin (TPP-4 COOH). The degradable flexible chain in the comonomer can effectively push away the porphyrin monomer with a rigid plane, so that the porphyrin monomer with the rigid plane can present a twisted non-planar structure in space, and the pi-pi interaction between molecules of the porphyrin monomer is greatly destroyed, thereby inhibiting the aggregation effect between porphyrin molecules, and simultaneously inhibiting the phenomena of low encapsulation rate, self-quenching and the like caused by the aggregation effect. Because the polyporphyrin has the characteristic of zwitterion, the polyporphyrin can be dissolved into purple liquid in water and self-assembled into particles with the particle size of 100-200nm through electrostatic action, and the singlet oxygen yield is improved by about 1.5 times compared with that of a monomer. Through the modification of surface polyethylene glycol (PEG) or Hyaluronic Acid (HA), the stability of the particles in PBS buffers with different pH values is obviously improved. The invention simultaneously solves the problems of aggregation induction quenching of hydrophobic porphyrin drugs, low drug-loading rate of nano-drugs and the like, provides a theoretical basis for the design of novel non-carrier photosensitizer drugs, and also provides a new idea for photodynamic tumor treatment.

Disclosure of Invention

The invention mainly solves the self-quenching effect caused by the aggregation of hydrophobic porphyrin drugs and simultaneously solves the problem of low drug loading of photosensitizer nano-drugs. Provides a preparation method of a novel polyporphyrin compound with a space distortion planar structure.

The technical scheme of the invention is as follows:

a preparation method of a polyporphyrin compound comprises the following steps:

dissolving tetraphenylporphyrin modified by tetracarboxylic acid in an organic solvent, wherein the concentration of the tetraphenylporphyrin is 0.1-1M; adding a thioketal comonomer with terminal diamino, finally adding 1-ethyl- (3-dimethylaminopropyl) carbonyldiimine hydrochloride and 4-dimethylaminopyridine into the reaction system, wherein the molar ratio of the tetra-carboxylic acid modified tetraphenylporphyrin, the thioketal comonomer with terminal diamino, 1-ethyl- (3-dimethylaminopropyl) carbonyldiimine hydrochloride and 4-dimethylaminopyridine is 1: 2.5-1.6: 4.8-8: 0.8-8, wherein the ratio of carboxylic acid to amino in the comonomer is 0.8-1.2; heating the reaction system to 50-100 ℃, and reacting for 24-72 h; after the reaction is finished, cooling to room temperature, then carrying out sedimentation centrifugation for 3-10 times by using a sedimentation agent, putting the crude product in a dialysis bag with the molecular weight cutoff of 100-1000, dialyzing for 3-5 days by using deionized water, and freeze-drying to obtain the polyporphyrin compound pPS, wherein the reaction formula is as follows:

tetracarboxylic acid modified tetraphenylporphyrin terminal diamino thioketal comonomer pPS

The polyporphyrin compound pPS is characterized in that:

the number average molecular weight is 7000 to 12000;

zeta potential: 7.2mV to-3.5 mV;

the molecular weight distribution PDI is 1.36-1.87.

The organic solvent is N, N-dimethylformamide or tetrahydrofuran.

The settling agent is diethyl ether or n-hexane.

The polyporphyrin compound is used for tumor photodynamic therapy.

The invention has the beneficial effects that: in the invention, water-soluble polyporphyrin is prepared by gradual polymerization, and compared with monomer porphyrin, the polyporphyrin can be self-assembled into nanoparticles under the electrostatic action in water. By mixing with polyethylene glycol or hyaluronic acid, a material with a stable nanostructure can be prepared, which has improved singlet oxygen yield and phototoxicity and can be used for the subsequent photodynamic treatment of malignant tumors.

Drawings

FIG. 1 is a nuclear magnetic characterization (d-DMSO) of the polymer (pPS).

FIG. 2 is a nuclear magnetic characterization of the photodegradation of polymer (pPS) (d-DMSO).

FIG. 3 is a scanning electron microscope image of the self-assembly of the polyporphyrin photosensitizers (PEG-pPS and HA-pPS). Wherein, a is PEG-pPS material, and b is HA-pPS material.

FIG. 4 is a representation of particle stability after self-assembly of the polyporphyrin photosensitizers (PEG-pPS and HA-pPS). Wherein, a is the particle size of the particles under different pH values, and b is the particle size of the particles under different dilution times.

FIG. 5 is the level of endocytosis of material in MCF-7 cells.

Figure 6 is an evaluation of the toxicity of the material. Wherein, a is the singlet oxygen yield of the polyporphyrin and the monomeric porphyrin, and b is the phototoxicity evaluation of the material in MCF-7 cells and H22 cells.

Detailed Description

The following further describes the specific embodiments of the present invention with reference to the technical solutions and the accompanying drawings.