阅读说明：本技术 一种pH敏感脂质材料及其制备方法和用途 (pH-sensitive lipid material and preparation method and application thereof ) 是由 徐缓 杜为昂 朱彩丽 王凯乾 于 2019-11-29 设计创作，主要内容包括：本发明属于有机化学合成、生物医药材料技术领域,公开了一种pH敏感脂质材料及其制备方法和用途。本发明提供的pH敏感脂质材料以2-甲基-2-噁唑啉为单体通过阳离子开环聚合反应合成一端为羟基的聚(2-甲基-2-噁唑啉),并将其与胆固醇甲酰氯酯化偶联合成制得。利用本发明制备的pH敏感脂质材料修饰的脂质体包封率高,稳定性好,表现出一定的pH敏感性。本发明的反应起始物价格低廉,反应条件温和,收率高,适合工业化生产。(The invention belongs to the technical field of organic chemical synthesis and biomedical materials, and discloses a pH sensitive lipid material and a preparation method and application thereof. The pH-sensitive lipid material provided by the invention is prepared by synthesizing poly (2-methyl-2-oxazoline) with one end being hydroxyl by taking 2-methyl-2-oxazoline as a monomer through a cationic ring-opening polymerization reaction, and performing esterification coupling on the poly (2-methyl-2-oxazoline) and cholesterol formyl chloride. The liposome modified by the pH sensitive lipid material prepared by the invention has high entrapment rate, good stability and certain pH sensitivity. The invention has the advantages of low price of reaction starting materials, mild reaction conditions and high yield, and is suitable for industrial production.)

1. A pH-sensitive lipid material is characterized by being a compound E shown as a general formula (I):

2. the method for preparing the pH-sensitive lipid material according to claim 1, wherein a compound A represented by a general formula (II) is used as a raw material, a compound B represented by a general formula (III) is used as an initiator, a polymerization reaction is carried out in an organic solvent to obtain a compound C represented by a general formula (IV), and the compound C represented by the general formula (IV) and a compound D represented by a general formula (V) are subjected to esterification coupling to finally obtain a compound E represented by a general formula (I);

wherein compound a: the mass ratio of the compound B is 5-100: 1, and the compound C: the mass ratio of the compound D is 1:1 to 10.

3. The method for preparing the pH-sensitive lipid material according to claim 2, wherein the compound C is prepared by: dissolving the compound A in an organic solvent under the condition of stirring, and mixing the compound A: adding the compound B into the mixture according to the proportion of 5-100: 1, heating and stirring the mixture at 50-120 ℃ for 12-72 hours, cooling the mixture to room temperature, adding a KOH methanol solution, stopping the reaction after 0.5-16 hours, filtering the product by silica gel, and then dropwise adding the product into diethyl ether to obtain a compound C.

4. The method for preparing the pH-sensitive lipid material according to claim 2, wherein the compound E is prepared by: dissolving the compound C in an organic solvent, adding a catalyst under the stirring condition, adding triethanolamine under the protection of nitrogen, and slowly dropwise adding an organic solution containing a compound D; stirring for 0.5-2 hours in ice bath at room temperature for 12-72 hours; pouring the crude product into a separating funnel, adding 0.01-1M hydrochloric acid, continuously washing for 2-5 times, then adjusting the pH to 5-8 with saturated sodium bicarbonate, washing with water, and extracting for 2-6 times; and (3) adding anhydrous magnesium sulfate into the product, and then separating and purifying by using column chromatography, wherein an eluent is a mixed solution of dichloromethane and methanol, and the volume ratio is 1-10: 1.

5. The method for preparing the pH sensitive lipid material according to claim 3, wherein in the preparation of the compound C, the organic solvent is any one or a combination of methanol, ethanol, diethyl ether, acetonitrile, chloroform, petroleum ether, acetone, NN-dimethylformamide and dimethyl sulfoxide.

6. The method for preparing the pH-sensitive lipid material according to claim 4, wherein in the preparation of the compound E, the catalyst is: pyridine, DMF, DMAP and aluminum trichloride or a combination of a plurality of pyridine, DMF, DMAP and aluminum trichloride.

7. The method for preparing the pH sensitive lipid material according to claim 4, wherein in the preparation of the compound E, the organic solvent is any one or a combination of several of dichloromethane, chloroform, carbon tetrachloride, bromobenzene and carbon disulfide.

8. The use of the pH sensitive lipid material compound E according to claim 1 in the preparation of an anti-tumor pharmaceutical preparation.

Technical Field

The invention belongs to the technical field of organic chemical synthesis and biomedical materials, and relates to a pH sensitive lipid material and a preparation method and application thereof.

Background

Cancer, which is a disease of color change in people, is characterized by a high morbidity and mortality, and is very alarming. Thus, many people gradually enter the "cancer prevention army", but nevertheless, cancer is still highly developed globally.

Official journal of the american cancer society published the "2018 global cancer statistics" report, which assesses the morbidity and mortality of 36 cancers in 185 countries.

In 2018, 1810 ten thousand new cancer cases and 960 ten thousand cancer death cases are predicted globally. This is the estimation data of the incidence of all cancers including non-melanoma skin cancers, all ages and sexes worldwide. Compared with other countries, the incidence and mortality of cancer are the first global! In 1810 ten thousands of new cancer cases, 380.4 ten thousands of cases exist in China; of 960 ten thousand cases of cancer deaths, 229.6 million cases are accounted for in our country.

This set of data implies: in every new 100 cancer patients worldwide, there are 21 Chinese people. That is, in China, there are 7 people diagnosed with cancer on average per minute, and nearly 5 people die of cancer per minute! In China, the incidence rate of lung cancer is the highest, so that the research on cancer treatment is urgent.

Traditional cancer treatment modalities include: surgery, radiation therapy and chemotherapy. Although these methods have some effects on cancer, it is known that surgical resection is quite painful for cancer patients and has some risks, but both radiotherapy and chemotherapy have the disadvantages of low bioavailability, great toxic and side effects, and the like. With the development of scientific technology, researchers are working on developing novel drug delivery systems to increase the availability of anticancer drugs in the body and reduce toxic side effects. The liposome consists of phospholipid and cholesterol, the structure of the liposome comprises a polar hydrophilic head group and two nonpolar carbon chains with hydrophobic effect, the hydrophilic part is exposed in the water phase, and the hydrophobic part avoids the water phase to form a complete closed vesicle. The structure of the carrier is similar to that of a biological membrane, so the carrier can be degraded in a living body as a drug delivery carrier, has low immunogenicity, can prolong the half-life period of the drug in the body and reduces toxic and side effects. Compared with normal cells, tumor cells are in an anoxic environment and glycolysis actively generates a large amount of acid products, and on the other hand, the over-expression of Carbonic Anhydrase (CA) by the tumor cells promotes CO₂And H₂O reaction generates a large amount of carbonic acid, which causes the tumor site to be slightly acidic, so how to trigger liposome to release the drug by using low pHBecomes a technical problem faced by scientific researchers.

Disclosure of Invention

In order to overcome the defects of the prior art, the first object of the invention is to provide a method for preparing liposome pH-sensitive lipid material, which provides possibility for constructing acid-sensitive liposomes. The second purpose of the invention is to provide the preparation method of the pH sensitive lipid material, which has the advantages of mild conditions, high yield, convenient operation and suitability for industrial mass production.

The above purpose of the invention is realized by the following technical scheme:

a pH-sensitive lipid material is a compound E shown in a general formula (I):

the preparation method of the pH-sensitive lipid material comprises the steps of taking a compound A shown in a general formula (II) as a raw material, taking a compound B shown in a general formula (III) as an initiator, carrying out polymerization reaction in an organic solvent to obtain a compound C shown in a general formula (IV), carrying out esterification coupling on the compound C shown in the general formula (IV) and a compound D shown in a general formula (V), and finally obtaining a compound E shown in a general formula (I) of the pH-sensitive lipid material.

Wherein compound a: the mass ratio of the compound B is 5-100: 1, and the compound C: the mass ratio of the compound D is 1: 1-10.

Preparation of compound C: dissolving the compound A in an organic solvent under the condition of stirring, and mixing the compound A: adding the compound B into the mixture according to the proportion of 5-100: 1, heating and stirring the mixture at 50-120 ℃ for 12-72 hours, cooling the mixture to room temperature, adding a KOH methanol solution, stopping the reaction after 0.5-16 hours, filtering the product by silica gel, and then dropwise adding the product into diethyl ether to obtain a compound C with the polymerization degree ranging from 5-100.

Preparation of compound E: dissolving the compound C in an organic solvent, adding a catalyst for acylation under the stirring condition, adding triethanolamine under the protection of nitrogen, and slowly dropwise adding an organic solution containing the compound D; stirring for 0.5-2 hours in ice bath at room temperature for 12-72 hours; pouring the crude product into a separating funnel, adding 0.01-1M hydrochloric acid, continuously washing for 2-5 times, then adjusting the pH to 5-8 with saturated sodium bicarbonate, and finally washing with water and extracting for 2-6 times; and (3) adding anhydrous magnesium sulfate into the product, and then separating and purifying by using column chromatography, wherein an eluent is a mixed solution of dichloromethane and methanol, and the volume ratio is 1-10: 1.

Further, in the preparation of the compound C, the organic solvent is any one or a combination of several of methanol, ethanol, diethyl ether, acetonitrile, chloroform, petroleum ether, acetone, NN-dimethylformamide and dimethyl sulfoxide.

Further, in the preparation of the compound E, the catalyst is: pyridine, DMF, DMAP and aluminum trichloride or a combination of a plurality of pyridine, DMF, DMAP and aluminum trichloride.

Further, in the preparation of the compound E, the organic solvent is any one or a combination of several of dichloromethane, chloroform, carbon tetrachloride, bromobenzene and carbon disulfide.

Further, the molecular weight range of the compound C is 500-10000.

The compound E can be applied to the preparation of antitumor pharmaceutical preparations.

Compared with the prior art, the invention has the beneficial effects that:

the compound A is a five-membered heterocyclic compound, and is a compound C obtained by cation ring-opening polymerization, wherein the compound C is a long-chain structure consisting of repeating units containing carbon and other atoms, and the hydroxyl at the tail end of the long chain shows that the compound C has stronger reaction activity. The invention connects the compound C with the compound D through electrophilic substitution reaction to obtain a novel polymer compound E. The polymer compound E prepared by the invention has high yield, narrow molecular weight distribution and good stability, can prevent the burst release of the drug and has obvious anticancer effect. The pH-sensitive lipid material provided by the invention is prepared by synthesizing poly (2-methyl-2-oxazoline) with one end being hydroxyl by taking 2-methyl-2-oxazoline as a monomer through a cationic ring-opening polymerization reaction, and performing esterification coupling on the poly (2-methyl-2-oxazoline) and cholesterol formyl chloride. The liposome modified by the pH sensitive lipid material has high entrapment rate and good stability, and shows certain pH sensitivity. The invention has the advantages of low price of reaction starting materials, mild reaction conditions and high yield, and is suitable for industrial production.

Drawings

FIG. 1 is an infrared spectrum of Compound C of example 1.

FIG. 2 is a drawing of Compound C of example 1¹H-NMR chart.

FIG. 3 is an infrared spectrum of Compound E of example 1.

FIG. 4 is a drawing of Compound E from example 1¹H-NMR chart.

Figure 5 is a graph of the percent drug leakage for different modified liposomes.

Detailed Description

The invention is described in more detail below with reference to specific examples, without limiting the scope of the invention. Unless otherwise specified, the experimental methods adopted by the invention are all conventional methods, and experimental equipment, materials, reagents and the like used in the experimental method can be obtained from commercial sources.