阅读说明：本技术 一种内镜粘膜下注射***剂的制备方法及其工艺 (Preparation method and process of endomucosal injection swelling agent ) 是由 曹加花 顾建军 张堃 陈雄伟 于 2020-06-12 设计创作，主要内容包括：本发明涉及一种内镜粘膜下注射隆起剂的制备方法及其工艺。它是由交联透明质酸钠和染色剂构成,其中交联透明质酸钠的质量含量为0.1%~0.8%,染色剂的的质量含量为0.0005%~0.05%；制备工艺是将透明质酸钠水溶液,加入氢氧化钠溶液搅拌使之变稀,再加入交联剂1,4-丁二醇二缩水甘油醚(BDDE)进行交联反应；交联结束后用盐酸溶液调整pH值,并用含染色剂的磷酸盐缓冲溶液稀释,均质后经湿热灭菌得到内镜粘膜下注射隆起剂。根据本发明制备的隆起剂,具有具有原料用量少,注入压力小,隆起高度高,维持时间长,可以终端灭菌,易于生产、运输和储存,更为安全的优点。(The invention relates to a preparation method and a process for injecting a swelling agent under an endoscope mucosa. The dyeing agent is composed of cross-linked sodium hyaluronate and a dyeing agent, wherein the mass content of the cross-linked sodium hyaluronate is 0.1-0.8%, and the mass content of the dyeing agent is 0.0005-0.05%; the preparation process comprises adding sodium hyaluronate aqueous solution into sodium hydroxide solution, stirring to thin, adding cross-linking agent 1, 4-butanediol diglycidyl ether (BDDE), and performing cross-linking reaction; and after the crosslinking is finished, adjusting the pH value by using a hydrochloric acid solution, diluting by using a phosphate buffer solution containing a coloring agent, homogenizing, and performing moist heat sterilization to obtain the endoscopic submucosal injection swelling agent. The swelling agent prepared according to the invention has the advantages of less raw material consumption, small injection pressure, high swelling height, long maintaining time, terminal sterilization, easy production, transportation and storage and higher safety.)

1. An endoscopic submucosa injection swelling agent comprises cross-linked sodium hyaluronate and a coloring agent, wherein the mass percentage concentration of the cross-linked sodium hyaluronate is 0.1-0.8%, and the mass percentage concentration of the coloring agent is 0.0005-0.05%; the preparation process comprises adding sodium hyaluronate aqueous solution into sodium hydroxide solution, stirring to thin, adding cross-linking agent 1, 4-butanediol diglycidyl ether (BDDE), and performing cross-linking reaction; adjusting the pH value with a hydrochloric acid solution after crosslinking is finished, diluting with a phosphate buffer solution containing a coloring agent, homogenizing, and performing moist heat sterilization to obtain an endoscopic submucosal injection swelling agent; the mass ratio of the cross-linking agent to the sodium hyaluronate is as follows: 0.001-0.1: 1.

2. An endoscopic submucosal injection swelling agent according to claim 1, wherein said sodium hyaluronate has a molecular weight of 20 to 2000kDa, preferably 150 to 1500 kDa.

3. The endoscopic submucosal injection swelling agent according to claim 1, wherein said cross-linking agent is replaced with a carbodiimide-based cross-linking agent, a sulfone-based cross-linking agent, an aldehyde-based cross-linking agent, and other epoxy-based cross-linking agents.

4. The endoscopic submucosal injection bump according to claim 1, wherein said coloring agent is replaced with indigo carmine or congo red.

5. The endomydrical injection bump agent of claim 1, characterized in that the sodium hyaluronate is coated with a polysaccharide: sodium alginate, cellulose and derivatives, starch, dextran, and chondroitin sulfate.

6. The method of claim 1, comprising the steps of:

1) dissolving sodium hyaluronate in water according to a certain amount to prepare an aqueous solution, and standing for 2-10 hours at the temperature of 10-30 ℃;

2) adding sodium hydroxide solution into the aqueous solution, and stirring to dilute the sodium hyaluronate aqueous solution;

3) adding a cross-linking agent into the diluted sodium hyaluronate aqueous solution, and stirring for reaction at the reaction temperature of 10-50 ℃ for 18-24 hours;

4) firstly, adjusting the pH value of a reaction system to 6.0-8.0 by using a hydrochloric acid solution, and then diluting by using a phosphate buffer solution containing a coloring agent to prepare a cross-linked gel with the mass percent of 0.1-0.8%;

5) and (3) homogenizing the cross-linked gel, and sterilizing at the temperature of 121 +/-1 ℃ for 5-15 min to obtain the endoscopic submucosal injection swelling agent.

7. The preparation method according to claim 6, wherein the mass percent concentration of the sodium hyaluronate is 0.5% -50%.

8. The preparation method according to claim 6, wherein the concentration of the sodium hydroxide solution is 2-20% by mass.

9. The preparation method according to claim 6, wherein the mass ratio of the cross-linking agent to the sodium hyaluronate is as follows: 0.001-0.1: 1.

10. The method according to claim 6, wherein the concentration of the coloring agent is 0.0005% to 0.05% by mass.

Technical Field

The invention relates to a preparation method and a process for injecting a swelling agent under an endoscope mucosa. The endoscopic submucosa injection swelling agent is a gel prepared from cross-linked polysaccharide, a coloring agent and a phosphate buffer solution, and can be used as a material for submucosal injection of the digestive tract.

Background

The digestive tract tumor accounts for more than half of the total malignant tumor in China, the early diagnosis and early treatment of the digestive tract tumor are the key points for improving the curative effect, and the digestive tract early cancer is treated by an endoscope, so that the survival rate in 5 years can be greatly improved to more than 90 percent, and the endoscope has the advantages of small wound, short hospitalization time, low cost and the like, thereby having an important role in diagnosing and treating the digestive tract early cancer and precancerous lesion.

In recent years, endoscopic techniques, such as endoscopic submucosal resection (EMR) and Endoscopic Submucosal Dissection (ESD), have revolutionized the diagnosis and treatment of digestive tract diseases and become the first treatment for early cancers and precancerous lesions of the digestive tract, and thus endoscopic submucosal injection of a swelling agent is particularly important in digestive endoscopic surgery. By injecting the submucosal swelling agent, the target mucosa can be swelled to facilitate excision, complications such as bleeding and perforation can be reduced, and whether endoscopic treatment can be performed can be preliminarily determined. Compared with EMR, the ESD excision range is larger, the operation time is correspondingly prolonged, the probability of complications in the treatment process is correspondingly increased due to the fact that the excision focus is larger, and effective submucosal injection is the decisive factor for ensuring ESD safety. Therefore, with the development of endoscopic technology, finding an ideal submucosal injection has become one of the targets that researchers and endoscopists have explored.

Sodium Hyaluronate (HA), a linear polysaccharide composed of double-bond repeating structural units formed by connecting D-glucuronic acid and N-acetyl-D-glucosamine through alternate (1 → 4) and (1 → 3) glycosidic bonds, is used as a human endogenous substance and is widely used for injection in multiple departments such as ophthalmology, orthopedics, plastic cosmetology and the like. Sodium hyaluronate has high viscosity and water absorption, and is not easy to be absorbed quickly after injection, so the swelling effect is superior to that of common clinical physiological saline, 50% glucose solution, hypertonic saline and concentrated glycerol-fructose solution.

Chinese patents CN105561402A and CN 105287626a disclose methods of using a non-crosslinked sodium hyaluronate solution as a submucosal injection for endoscopes, respectively, the swelling effect is significantly better than that of normal saline, and sodium hyaluronate with excellent lubricating effect has physiological functions of promoting angiogenesis, promoting wound healing, resisting tumor, regulating immunity, and the like.

CN107456612A also discloses a submucosal injection solution and the like. However, the sodium hyaluronate solution is not stable, has strict requirements on sterilization mode and storage conditions, needs to be produced by an aseptic processing method, and increases the difficulty for production, processing, transportation and storage of products. Because the cost of the sodium hyaluronate is higher, the reduction of the dosage of the sodium hyaluronate in the product is particularly critical.

Disclosure of Invention

The invention aims to provide a preparation method and a process of an endoscopic submucosal injection swelling agent, which can solve the technical problems. .

The endoscopic submucosa injection swelling agent provided by the invention is composed of cross-linked sodium hyaluronate and a coloring agent, wherein the mass percentage concentration of the cross-linked sodium hyaluronate is 0.1-0.8%, and the mass percentage concentration of the coloring agent is 0.0005-0.05%; the preparation process comprises adding sodium hyaluronate aqueous solution into sodium hydroxide solution, stirring to thin, adding cross-linking agent 1, 4-butanediol diglycidyl ether (BDDE), and performing cross-linking reaction; and after the crosslinking is finished, adjusting the pH value by using a hydrochloric acid solution, diluting by using a phosphate buffer solution containing a coloring agent, homogenizing, and performing moist heat sterilization to obtain the endoscopic submucosal injection swelling agent.

The mass ratio of the cross-linking agent to the sodium hyaluronate is as follows: 0.001-0.1: 1.

The cross-linking agent can be replaced by carbodiimide cross-linking agent, sulfone cross-linking agent, aldehyde cross-linking agent and other epoxy cross-linking agent.

The coloring agent can be replaced by indigo carmine and Congo red.

The molecular weight range of the sodium hyaluronate is 20-2000 kDa, and the preferred molecular weight range is 150-1500 kDa.

The sodium hyaluronate can be prepared from the following polysaccharides: sodium alginate, cellulose and its derivatives, starch, dextran, and chondroitin sulfate.

The preparation method of the endomucosal injection swelling agent provided by the invention comprises the following steps:

1) dissolving sodium hyaluronate in water according to a certain amount to prepare an aqueous solution, and standing and dissolving for 2-10 hours at the temperature of 10-30 ℃. The mass percentage concentration of the sodium hyaluronate is 0.5-50%.

2) Sodium hydroxide solution was added to the aqueous solution and stirred to thin the sodium hyaluronate aqueous solution. The mass percentage concentration of the sodium hydroxide solution is 2-20%.

3) And adding a cross-linking agent into the diluted sodium hyaluronate aqueous solution, and stirring for reaction at the reaction temperature of 10-50 ℃ for 18-24 hours. The mass ratio of the cross-linking agent to the sodium hyaluronate is as follows: 0.001-0.1: 1.

4) Firstly, adjusting the pH value of a reaction system to 6.0-8.0 by using a hydrochloric acid solution, and then diluting by using a phosphate buffer solution containing a coloring agent to prepare a cross-linked gel with the mass percent of 0.1-0.8%. The mass percentage concentration of the coloring agent is 0.0005% -0.05%.

5) And (3) homogenizing the cross-linked gel, and sterilizing at the temperature of 121 +/-1 ℃ for 5-15 min to obtain the endoscopic submucosal injection swelling agent.

The invention provides a preparation method and a process for an endoscopic submucosal injection swelling agent, which solve the problems in the prior art, adopts a chemical method to crosslink polysaccharide, and adds a coloring agent to obtain an endoscopic submucosal injection swelling agent gel.

The invention has the outstanding characteristics that after the polysaccharide is crosslinked to form gel, all molecules inside the gel are connected to form a net-shaped bracket, and based on the three-dimensional space grid, a large amount of water can be combined, so that the gel with extremely low concentration can achieve a better swelling effect, and the low concentration can reduce the consumption of raw materials and simultaneously keep lower injection pressure. Compared with the non-crosslinked sodium hyaluronate solution, the dosage of the raw materials can be reduced by more than 50%, the injection pressure can be reduced by more than 20%, and the bulge height after 60min can be improved by more than 14%.

Drawings

FIG. 1 is a graph showing the effect of comparative experiments on the elevation height at the initial time of injecting the present invention under the gastric mucosa of a rabbit; comparative example A: HA molecular weight is 825kD, concentration is 0.4%; example 4: the molecular weight of HA is 1270kD, the mass ratio of the cross-linking agent to the HA is 0.005:1, and the concentration is 0.4%; example 7: the molecular weight of HA is 825kD, the mass ratio of the cross-linking agent to the HA is 0.0075:1, and the concentration is 0.2%; example 9: the molecular weight of HA is 1270kD, the mass ratio of the cross-linking agent to HA is 0.0075:1, and the concentration is 0.2%).

FIG. 2 is a graph showing the effect of comparative experiments on the elevation height of a rabbit injected under the gastric mucosa for 60 min; (control example A: HA molecular weight 825kD at 0.4%; example B4: HA molecular weight 1270kD, crosslinker to HA mass ratio 0.005:1, concentration 0.4%; example C7: HA molecular weight 825kD, crosslinker to HA mass ratio 0.0075:1, concentration 0.2%; example D9: HA molecular weight 1270kD, crosslinker to HA mass ratio 0.0075:1, concentration 0.2%).

Detailed Description

The present invention is further illustrated by the following examples, which do not limit the scope of the present invention.