阅读说明：本技术 一种抗肿瘤活性蔗渣木聚糖肌醇六磷酸酯-g-BMA的合成方法 (Synthesis method of bagasse xylan phytate-g-BMA with antitumor activity ) 是由 李和平 葛文旭 李明坤 杨锦武 武晋雄 杨莹莹 耿恺 张淑芬 郑光绿 于 2019-10-22 设计创作，主要内容包括：本发明公开了一种抗肿瘤活性蔗渣木聚糖肌醇六磷酸酯-g-BMA的合成方法。以蔗渣木聚糖为主要原料,甲基丙烯酸苄酯(BMA)为接枝单体,以过硫酸钾为引发剂,在水溶剂中合成蔗渣木聚糖-g-BMA的基础上,再以过硫酸铵与一水合次亚磷酸钠为复合催化剂,肌醇六磷酸为酯化剂,在离子液体氯化-1-烯丙基-3-甲基咪唑内经复合催化酯化反应合成了蔗渣木聚糖肌醇六磷酸酯-g-BMA。本发明经过接枝、酯化两步反应合成具有生物活性的蔗渣木聚糖肌醇六磷酸酯-g-BMA。在引入多种活性基团的共同作用下,蔗渣木聚糖的水溶性、热稳定性、生物活性得到显著改善,其抗肿瘤活性也明显增强,产物在医药领域具有广阔的应用前景。(The invention discloses a method for synthesizing anti-tumor active bagasse xylan phytate-g-BMA. Bagasse xylan is used as a main raw material, Benzyl Methacrylate (BMA) is used as a grafting monomer, potassium persulfate is used as an initiator, on the basis of synthesizing the bagasse xylan-g-BMA in a water solvent, ammonium persulfate and sodium hypophosphite monohydrate are used as a composite catalyst, phytic acid is used as an esterifying agent, and the bagasse xylan phytate-g-BMA is synthesized through a composite catalytic esterification reaction in ionic liquid chloro-1-allyl-3-methylimidazole. The bagasse xylan phytate-g-BMA with biological activity is synthesized through two-step reactions of grafting and esterification. Under the combined action of introducing various active groups, the water solubility, the thermal stability and the biological activity of the bagasse xylan are obviously improved, the anti-tumor activity of the bagasse xylan is also obviously enhanced, and the product has wide application prospects in the field of medicines.)

1. A method for synthesizing anti-tumor active bagasse xylan phytate-g-BMA is characterized by comprising the following specific steps:

(1) placing 20-25 g of bagasse xylan into a vacuum constant-temperature drying oven at 50 ℃ for drying for 24 hours to obtain dry-based bagasse xylan;

(2) weighing 3-4 g of potassium persulfate in a 50mL beaker, adding 20-30 mL of deionized water, stirring at room temperature for 5-8 minutes to obtain an initiator solution, and pouring the initiator solution into a 100mL constant-pressure dropping funnel for later use;

(3) weighing 15-20 g of the dry-based bagasse xylan obtained in the step (1), placing the dry-based bagasse xylan into a 250mL four-neck flask provided with a stirrer, a thermometer and a reflux condensing device, adding 40-50 mL of deionized water, and stirring at room temperature for 15-20 minutes to obtain a bagasse xylan suspension;

(4) heating the system in the step (3) to 60-80 ℃, starting to dropwise add the initiator solution prepared in the step (2), simultaneously dropwise adding 15-20 mL of analytically pure Benzyl Methacrylate (BMA) monomer, and controlling the dropwise addition to be completed within 3-4 hours; after the monomer and the initiator solution are dripped, continuously reacting for 0.5-1 hour, and cooling the material to room temperature;

(5) adding 30-50 mL of analytically pure absolute ethyl alcohol into the material obtained in the step (4), uniformly stirring, standing for 30-40 minutes, filtering, washing with 20-30 mL of analytically pure absolute ethyl alcohol and 20-30 mL of analytically pure acetone in sequence, and performing suction filtration for 2-3 times to obtain a bagasse xylan-g-BMA crude product;

(6) putting the bagasse xylan-g-BMA crude product obtained in the step (5) into a Soxhlet extractor, and adding 150-200 mL of cyclohexane to extract for 24 hours to remove a homopolymer; after extraction, taking out the materials, putting the materials into a watch glass, and drying the materials in a vacuum constant-temperature drying oven at 50 ℃ for 24 hours to obtain purified graft copolymer bagasse xylan-g-BMA;

(7) weighing 40-45 mL of analytically pure allyl chloride and 30-35 mL of analytically pure 1-methylimidazole, placing the analytically pure allyl chloride and the 1-methylimidazole into a 250mL round-bottom flask sealing system provided with a stirrer, a reflux condensing device and a thermometer, controlling the temperature to be 55-60 ℃, and stirring and reacting the system for 7-9 hours under a continuous vacuum-pumping state by adopting a circulating water type vacuum pump; evaporating excessive allyl chloride in the obtained crude product by using a rotary evaporator to obtain ionic liquid 1-allyl-3-methylimidazole chloride, cooling the ionic liquid to room temperature, and pouring the ionic liquid into a 100mL beaker;

(8) weighing 30-40 g of ionic liquid AmimCl obtained in the step (7) as an esterification reaction solvent, placing the esterification reaction solvent into a 250mL four-neck flask provided with a stirrer, a thermometer and a reflux condensing device, and then adding 12-15 g of bagasse xylan-g-BMA obtained in the step (6); heating the system to 80-95 ℃, and stirring for 40-60 minutes; then adding 1-2 g of ammonium persulfate and 1.2-2.5 g of sodium hypophosphite monohydrate, and simultaneously dropwise adding 40-50 mL of phytic acid aqueous solution containing 50% of mass fraction, wherein the dropwise adding is controlled to be finished within 3-4 hours; continuing to react for 0.5-1 hour, and cooling the material to room temperature;

(9) adding 30-50 mL of analytically pure absolute ethyl alcohol into the material obtained in the step (8), uniformly stirring, standing for 30-50 minutes, filtering, washing with 20-30 mL of analytically pure absolute ethyl alcohol and 20-30 mL of analytically pure acetone respectively in sequence, and performing suction filtration for 2-3 times; and putting the filter cake into a surface dish, and drying in a vacuum constant-temperature drying oven at 60 ℃ for 24 hours to obtain the bagasse xylan phytate-g-BMA.

Technical Field

The invention relates to the technical field of fine chemical engineering, in particular to a method for synthesizing anti-tumor active bagasse xylan phytate-g-BMA.

Background

The treatment of tumors has been a worldwide problem, and malignant tumors remain a major disease that endangers human health. The tumor treatment difficulty is high due to poor tumor treatment effect, high recurrence and metastasis rate, large side effect of tumor treatment, poor accuracy and the like. Researches show that the xylan has good biocompatibility and biological activities of tumor resistance, immunoregulation and the like, and provides possibility for the application of the xylan in the aspect of antitumor drugs. At present, the research on natural antitumor drugs such as paclitaxel and camptothecin is relatively more, and the research on the antitumor aspect of xylan is only reported in documents.

The xylan grafted esterified derivative can effectively inhibit the diffusion of tumor cells, avoid the damage to normal cells and induce the apoptosis of the tumor cells more purposefully. Benzyl Methacrylate (BMA) is used as a grafting monomer to carry out graft copolymerization with the bagasse xylan, so that the water solubility and the thermal stability of the bagasse xylan can be improved, and the anti-tumor activity of the bagasse xylan can be obviously enhanced. The phytic acid has physiological functions of resisting oxidation, resisting tumor and the like, and can obviously inhibit the proliferation of cancer cells. The bagasse xylan-g-BMA is further subjected to esterification modification by using phytic acid as an esterifying agent, so that the biological activity of the bagasse xylan can be further enhanced, and the targeting anti-tumor effect can be better exerted.

The bagasse xylan is used as a main raw material, Benzyl Methacrylate (BMA) is used as a grafting monomer, potassium persulfate is used as an initiator, on the basis of synthesizing the bagasse xylan-g-BMA in a water solvent, ammonium persulfate and sodium hypophosphite monohydrate are used as a composite catalyst, phytic acid is used as an esterifying agent, and the bagasse xylan phytate-g-BMA is synthesized through a composite catalytic esterification reaction in ionic liquid chloro-1-allyl-3-methylimidazole (AmimCl).

Disclosure of Invention

The invention aims to enhance the anti-tumor activity of bagasse xylan, develop and utilize the medicinal value of bagasse xylan, and provide a synthesis method of bagasse xylan phytate-g-BMA.

The method comprises the following specific steps:

(1) and (3) drying 20-25 g of bagasse xylan in a vacuum constant-temperature drying oven at 50 ℃ for 24 hours to obtain the dry-based bagasse xylan.

(2) Weighing 3-4 g of potassium persulfate in a 50mL beaker, adding 20-30 mL of deionized water, stirring at room temperature for 5-8 minutes to obtain an initiator solution, and pouring the initiator solution into a 100mL constant-pressure dropping funnel for later use.

(3) Weighing 15-20 g of the dry bagasse xylan obtained in the step (1), placing the dry bagasse xylan into a 250mL four-neck flask provided with a stirrer, a thermometer and a reflux condensing device, adding 40-50 mL of deionized water, and stirring at room temperature for 15-20 minutes to obtain a bagasse xylan suspension.

(4) And (3) heating the system in the step (3) to 60-80 ℃, starting to dropwise add the initiator solution prepared in the step (2), simultaneously dropwise adding 15-20 mL of analytically pure benzyl methacrylate, namely BMA monomer, and controlling the dropwise addition to be finished within 3-4 hours. And after the monomer and the initiator solution are dripped, continuously reacting for 0.5-1 hour, and cooling the material to room temperature.

(5) And (3) adding 30-50 mL of analytically pure absolute ethyl alcohol into the material obtained in the step (4), uniformly stirring, standing for 30-40 minutes, filtering, washing with 20-30 mL of analytically pure absolute ethyl alcohol and 20-30 mL of analytically pure acetone in sequence, and performing suction filtration for 2-3 times to obtain a bagasse xylan-g-BMA crude product.

(6) And (3) putting the bagasse xylan-g-BMA crude product obtained in the step (5) into a Soxhlet extractor, and adding 150-200 mL of cyclohexane to extract for 24 hours to remove the homopolymer. And (3) taking out the material after extraction, putting the material into a watch glass, and drying the material in a vacuum constant-temperature drying oven at 50 ℃ for 24 hours to obtain the purified graft copolymer bagasse xylan-g-BMA.

(7) Weighing 40-45 mL of analytically pure allyl chloride and 30-35 mL of analytically pure 1-methylimidazole, placing the analytically pure allyl chloride and the 1-methylimidazole into a 250mL round-bottom flask sealing system provided with a stirrer, a reflux condensing device and a thermometer, controlling the temperature to be 55-60 ℃, and stirring and reacting the system for 7-9 hours under a continuous vacuum-pumping state by adopting a circulating water type vacuum pump. Evaporating excessive allyl chloride in the obtained crude product by a rotary evaporator to obtain ionic liquid 1-allyl-3-methylimidazole chloride, cooling the ionic liquid to room temperature, and pouring the ionic liquid into a 100mL beaker.

(8) And (3) weighing 30-40 g of the ionic liquid AmimCl obtained in the step (7) as an esterification reaction solvent, placing the esterification reaction solvent into a 250mL four-neck flask provided with a stirrer, a thermometer and a reflux condensing device, and then adding 12-15 g of the bagasse xylan-g-BMA obtained in the step (6). Heating the system to 80-95 ℃, and stirring for 40-60 minutes. Then adding 1-2 g of ammonium persulfate and 1.2-2.5 g of sodium hypophosphite monohydrate; beginning to dropwise add 40-50 mL of phytic acid aqueous solution with the mass fraction of 50%, and controlling the dropwise adding to be finished within 3-4 hours; and continuing to react for 0.5-1 hour, and cooling the material to room temperature.

(9) And (3) adding 30-50 mL of analytically pure absolute ethyl alcohol into the material obtained in the step (8), uniformly stirring, standing for 30-50 minutes, filtering, washing with 20-30 mL of analytically pure absolute ethyl alcohol and 20-30 mL of analytically pure acetone respectively in sequence, and performing suction filtration for 2-3 times. And putting the filter cake into a surface dish, and drying in a vacuum constant-temperature drying oven at 60 ℃ for 24 hours to obtain the bagasse xylan phytate-g-BMA.

(10) The phosphorus content is measured by adopting a phosphorus-molybdenum blue method of GB/T5750.5-2006, and the substitution degree is calculated by the following formula:

in the formula:

31-phytic acid acyl relative molecular mass;

132-relative molecular mass of bagasse xylan dewatering units;

W_p-the phosphorus content in the sample,%;

DS-degree of substitution.

The bagasse xylan phytate-g-BMA with biological activity is synthesized through two-step reactions of grafting and esterification. Under the combined action of introducing various active groups, the water solubility, the thermal stability and the biological activity of the bagasse xylan are obviously improved, the anti-tumor activity of the bagasse xylan is also obviously enhanced, and the product has wide application prospects in the field of medicines.

Drawings

FIG. 1 is an SEM photograph of raw bagasse xylan.

FIG. 2 is an SEM photograph of bagasse xylan phytate-g-BMA.

FIG. 3 is an IR chart of crude bagasse xylan and bagasse xylan phytate-g-BMA.

Figure 4 is an XRD pattern of raw bagasse xylan.

FIG. 5 is an XRD pattern of bagasse xylan phytate-g-BMA.

FIG. 6 is a graph showing TG and DTG curves of raw bagasse xylan.

FIG. 7 is a TG and DTG curve of bagasse xylan phytate-g-BMA.

Detailed Description