阅读说明：本技术 一种莱啉系基近红外材料的制备方法 (Preparation method of oxazoline-based near-infrared material ) 是由 周英棠 于 2019-08-07 设计创作，主要内容包括：本发明涉及制药领域,针对现有技术的肿瘤靶向治疗药物载体基因承载能力较弱和生物相容性差的问题,公开了一种莱啉系基近红外材料的制备方法,(1)、合成莱啉基单氯取代苝二酸酐化合物：a、将苝二酸酐溶于浓硫酸中,质量比为0.7-1.2：2.2-2.4；b、在混合溶液中缓慢滴加浓硫酸的TBCA溶液；c、当出现醇相,去离子水透析净化,期间避光,冷冻干燥后得到PDI；(2)、制备莱啉系基近红外材料纳米药物：将上述PDI与聚乙二醇以质量比为0.8-1:1.4-1.8溶解在去离子水中并超声搅拌处理。本发明具有较大的拓扑支链结构,较强的药物承载能力,及具备一定的杀菌能力,所得产物热稳定性较好,且制备流程简单。(The invention relates to the field of pharmacy, and discloses a preparation method of a Lelinyl-based near-infrared material aiming at the problems of weak gene bearing capacity and poor biocompatibility of a tumor targeted therapy drug carrier in the prior art, wherein the preparation method comprises the following steps of (1) synthesizing a Lelinyl monochloro substituted perylene dianhydride compound: a. dissolving perylene dianhydride in concentrated sulfuric acid, wherein the mass ratio is 0.7-1.2: 2.2-2.4; b. slowly dripping TBCA solution of concentrated sulfuric acid into the mixed solution; c. when an alcohol phase appears, dialyzing and purifying with deionized water, keeping out of the sun during the period, and freeze-drying to obtain PDI; (2) and preparing the oxazoline-based near-infrared material nano-drug: the PDI and the polyethylene glycol are dissolved in deionized water according to the mass ratio of 0.8-1:1.4-1.8 and are subjected to ultrasonic stirring treatment. The invention has larger topological branched chain structure, stronger drug bearing capacity and certain sterilization capacity, and the obtained product has better thermal stability and simple preparation process.)

1. A preparation method of a Leolin-based near-infrared material is characterized by comprising the following steps:

(1) synthesizing a Lelinyl monochloro substituted perylene dianhydride compound:

a. dissolving perylene dianhydride in concentrated sulfuric acid, wherein the mass ratio is 0.7-1.2: 2.2-2.4 until all the components are dissolved, and mixing uniformly;

b. slowly dripping TBCA solution of concentrated sulfuric acid into the mixed solution obtained in the step a, keeping reaction, and then detecting the reaction degree;

c. when an alcohol phase appears, separating a high molecular weight product by a chromatographic column, dialyzing and purifying by deionized water, keeping out of the sun during the dialysis and purification, and freeze-drying to obtain green powder, namely the Lelinyl monochloro substituted perylene dianhydride compound;

(2) and preparing the oxazoline-based near-infrared material nano-drug: dissolving the quantitative green powder PDI and polyethylene glycol in deionized water in a mass ratio of 0.8-1:1.4-1.8, and performing ultrasonic stirring treatment to obtain a finished product, and storing the finished product.

2. The method according to claim 1, wherein the step a is performed under magnetic stirring in a nitrogen atmosphere.

3. The method according to claim 1, wherein the concentrated sulfuric acid in step a has a volume concentration of 65-70%.

4. The method of claim 1, wherein the maintaining time in step b is 2.5 ~ 3 hours.

5. The method according to claim 1, wherein the step b is carried out at-1 ~ 0 ℃.

6. The method as claimed in claim 1, wherein the dialysis membrane cut-off molecular weight of the dialysis purification site in step c is Mw =2000-2500 Da.

7. The method according to claim 1, wherein the dialysis purification time in step c is 24-30 h.

8. The method for preparing a lyoline-based near-infrared material according to claim 1, wherein the storage temperature in the step (2) is 2 to 8 ℃.

9. The method according to claim 1, wherein the polyethylene glycol is modified polyethylene glycol, the modified polyethylene glycol is prepared by mixing polyethylene glycol and flavanone compound at a mass ratio of 4.8-5.0:1.3-1.6, heating and stirring at 90 ~ 110 ℃ for 2.5-3.5h at a stirring speed of 1000-.

10. The method of claim 9, wherein the concentration of diethyl ether is 95 ~ 98% by volume.

Technical Field

The invention relates to the field of pharmacy, in particular to a preparation method of a Leolin-based near-infrared material.

Background

Near-infrared light-excited photothermal therapy can effectively make up for the defects of the traditional cancer treatment means, but the currently commonly used near-infrared organic photothermal molecules (such as cyanine and the like) have poor light stability, and the light degradation of the near-infrared organic photothermal molecules can influence the stability of the nano structure and the photothermal conversion efficiency. Therefore, a novel near-infrared organic photothermal reagent with high stable nanostructure and high photothermal conversion efficiency is developed, so that the treatment efficiency and the medical safety of tumors are effectively improved. In addition, the infrared material is used as a carrier for in vivo tumor targeted therapy, so the antibacterial performance of the material is also essential.

The patent number CN201810246008.4 is named as 'a photo-thermal tumor drug and synthesis and application thereof in tumor treatment', and provides the photo-thermal tumor drug, which takes a phase-change material as a shell and is internally packaged with a target drug, a near-infrared probe and CuS nano particles. The experiment of the primary laser irradiation temperature rise curve of the photothermal tumor medicament proves that the photothermal material has higher photothermal conversion efficiency, and electron microscope and particle size analysis show that the photothermal material has the characteristic of passively targeting tumors by nano materials, and can be used for combined treatment of chemotherapy and thermotherapy of tumor parts.

The disadvantages are that the hydrophilicity, the biocompatibility and the gene bearing capacity are weak, and the antibacterial performance is poor.

Disclosure of Invention

The invention aims to overcome the problems of weak gene bearing capacity and poor biocompatibility of tumor targeted therapy drug carriers in the prior art, and provides a preparation method of a Leolin-based near-infrared material.

In order to achieve the purpose, the invention adopts the following technical scheme:

a preparation method of a Leolin-based near-infrared material comprises the following steps:

(1) synthesizing a Lelinyl monochloro substituted perylene dianhydride compound:

a. dissolving perylene dianhydride in concentrated sulfuric acid, wherein the mass ratio is 0.7-1.2: 2.2-2.4 until all the components are dissolved, and mixing uniformly;

b. slowly dripping TBCA solution of concentrated sulfuric acid into the mixed solution obtained in the step a, keeping reaction, and then detecting the reaction degree;

c. when an alcohol phase appears, separating a high molecular weight product by a chromatographic column, dialyzing and purifying by deionized water, keeping out of the sun during the dialysis and purification, and freeze-drying to obtain green powder, namely the Lelinyl monochloro substituted perylene dianhydride compound;

(2) and preparing the oxazoline-based near-infrared material nano-drug: dissolving the quantitative green powder PDI and polyethylene glycol in deionized water in a mass ratio of 0.8-1:1.4-1.8, and performing ultrasonic stirring treatment to obtain a finished product, and storing the finished product.

The perylene dianhydride is a carrier without biological toxicity, has higher dendritic molecular algebra, has a larger dendritic structure and more positive charges ionized by the tail end amino group, and has stronger gene loading capacity.

PDI (Lelinyl monochloro substituted perylene dianhydride compound) is an organic molecule with strong near-infrared energy absorption, can keep chemical structure stability in acid-base, strong light, high heat and other environments, and has the characteristics of high extinction coefficient and easiness in chemical modification. Therefore, a shell-core topological molecular structure with PDI as a molecular core is introduced to realize the multifunctional biological application of the perylene imide macromolecule, the shell structure is polyethylene glycol (PEG), and the PDI and the polyethylene glycol are subjected to esterification reaction to obtain the polyimide modified perylene imide macromolecule.

The polyethylene glycol has good water solubility, good intermiscibility with a plurality of organic matter components, good hygroscopicity, lubricity, cohesiveness and chemical stability.

Due to the hydrophobicity of polyimide and the hydrogen bond driving force of polyethylene glycol, the photo-thermal nano-drug with good biocompatibility is prepared by self-assembly of macromolecules in aqueous solution, and has stable photo-thermal performance and higher photo-thermal conversion efficiency.

Preferably, step a is carried out under magnetic stirring in a nitrogen atmosphere.

The nitrogen atmosphere can prevent the reactants from being oxidized, and the magnetic stirring is used for fully dissolving the perylene dianhydride and the concentrated sulfuric acid to better form a uniform mixed solution.

Preferably, the volume concentration of the concentrated sulfuric acid in the step a is 65-70%.

The perylene dianhydride cannot be well dispersed when the concentration of the concentrated sulfuric acid is too low, and the perylene dianhydride is easily oxidized when the concentration of the concentrated sulfuric acid is too high, so that a solution with the concentration required by the reaction cannot be obtained.

Preferably, the reaction time in step b is kept at 2.5 ~ 3 h.

Preferably, step b is carried out dropwise at-1 ~ 0 ℃.

Preferably, the dialysis membrane cut-off at said dialysis purification in step c is Mw =2000-2500 Da.

Preferably, the dialysis purification time in step c is 24-30 h.

Preferably, the storage temperature in step (2) is 2 to 8 ℃.

Preferably, the polyethylene glycol in the step (2) is modified polyethylene glycol, the modified polyethylene glycol is prepared by mixing the polyethylene glycol and the flavanone compound according to the mass ratio of 4.8-5.0:1.3-1.6, heating and stirring, wherein the heating temperature is 90 ~ 110 ℃, the heating time is 2.5-3.5h, the stirring speed is 1000-.

The modified polyethylene glycol has antibacterial property while ensuring the hydrophilic property, dispersibility and photo-thermal stability, and can prevent the surrounding part from being infected when the drug carrier is injected into the body, in the process of killing tumor cells or reaches the inflamed part.

The polyethylene glycol and the flavanone compound must have a proper mass ratio, excessive flavanone compound components can excessively consume hydrogen on a polyethylene glycol molecular chain, and if the proportion of the flavanone compound is too low, the antibacterial capability of the obtained modified polyethylene glycol is insufficient, so that an ideal antibacterial effect cannot be achieved.

Introducing the flavanone compound to carry out copolymerization reaction with polyethylene glycol, and carrying out reaction between 4-position hydroxyl on a molecular chain of the flavanone compound and hydrogen on a molecular chain of the polyethylene glycol to obtain modified polyethylene glycol with antibacterial property, wherein pentenyl substitutes on the molecular chain of the flavanone compound have stronger antibacterial and bactericidal capabilities.

The heating energy provides power for the copolymerization reaction to promote the reaction to occur, and the heating, the heat preservation for a period of time and the stirring at a certain stirring speed are carried out to ensure that the reaction is fully carried out, so as to obtain the polyethylene glycol with better antibacterial property.

Preferably, the concentration of diethyl ether is 95 ~ 98% by volume.

In order to ensure that the modified polyethylene glycol in the mixed solution can be fully precipitated and separated out, the component purity of the modified polyethylene glycol is ensured.

Therefore, the invention has the following beneficial effects:

(1) the nano-composite material has a larger topological branched chain structure, stronger drug carrying capacity, stronger photo-thermal performance and photo-thermal conversion rate;

(2) the antibacterial agent has stronger antibacterial capacity, and the obtained product has better thermal stability and stronger biocompatibility;

(3) the ultra-small, controllable and stable nanostructure is beneficial to deep penetration of tumor tissues and effective metabolism from the body, can penetrate deeper tissues and reduce the secondary damage to organisms caused by drug dosage and laser radiation.

Drawings

FIG. 1 is a graph showing cytotoxicity data of PDI-NPs of various concentrations according to the present invention (the left side of the graph represents 10% PDI-NP, the middle represents 5% PDI-NP, and the right side represents Control).

FIG. 2 is a graph showing the effect of photothermal therapy at the cellular level of PDI-NP at various concentrations according to the present invention.

Detailed Description

The invention is further described with reference to specific embodiments.