CN112076341A

CN112076341A - Self-curing self-adhesive digestive tract injury mucosa protective adhesive and application thereof

Info

Publication number: CN112076341A
Application number: CN201910448532.4A
Authority: CN
Inventors: 戴建英
Original assignee: Individual
Current assignee: Individual
Priority date: 2019-05-27
Filing date: 2019-05-27
Publication date: 2020-12-15

Abstract

The invention provides a self-curing self-adhesive protective adhesive product for digestive tract injured mucosa, belonging to the field of biological pharmaceutical preparations. The self-curing self-adhesive digestive tract injury mucosa protective glue is gel and biological adhesive; the effective components of the gel are gel forming substances, water-soluble calcium salt and a calcium ion complexing agent; the biological adhesive is polylysine; the pH value of the digestive tract injury mucosa protective gel is not less than 7. And curing liquid can be added to cure the protective glue at the designated position. The protective glue can form a colloid film on the surface of the ulcer to promote the healing of the ulcer. In addition, the curing liquid is adopted to ensure that the protective adhesive is rapidly fixed on the surface of the damaged mucosa, thereby avoiding the flowing problem of the protective adhesive. Animal experiments show that the invention can obviously promote the repair of the injured gastric mucosa, thereby having the effects of promoting healing and protecting the injury of the gastric mucosa.

Description

Self-curing self-adhesive digestive tract injury mucosa protective adhesive and application thereof

Technical Field

The invention relates to the technical field of biological medicines, in particular to a preparation method and a product of a self-curing self-adhesive digestive tract injury mucosa protective adhesive which is sprayed on the surface of the digestive tract injury mucosa through a gastrointestinal endoscope.

Background

The endoscopic gastric mucosa resection and gastric mucosa stripping are minimally invasive surgeries for resection of early gastric cancer and gastric benign tumors, can achieve the aim of radically treating the early gastric cancer, have the advantages of small wound, small influence on the life quality of patients and the like, and gradually replace part of the traditional surgical operations. Minimally invasive surgery for early cancerous and benign tumor removal of the esophagus and colon also replaces part of traditional surgery. However, at present, the wound surface after the operation is only subjected to hemostasis treatment, and no good wound surface protection measures are available. The naked wound surface is subjected to an ulcer active phase, a healing phase and a scar phase under the action of digestive juice, and the healing time of the wound surface is longer. Clinical research data show that the healing rate of patients after 6 weeks of endoscopic gastric mucosal resection and gastric mucosal dissection is about 69%. Therefore, there is a need to provide a product that can be applied directly to the mucosa surface of a gastric lesion via gastroscopy.

Alginate is used as excellent matrix of medicinal preparation, and is used as hydrophilic emulsifier, gel and thickener. Chinese patent application No.: 101933894B discloses a gastric mucosa protective gel, which comprises gel forming substance, acid-base regulator, cross linker and adhesive; the weight ratio of the cross-linking agent to the adhesive is 1: 0.01-0.3: 0.1-0.5, and the dosage of the acid-base regulator is to regulate the pH value to 6.5-8.5. The patent only utilizes the pure physical principle to protect the gastric mucosa, the used cross-linking agent is acid-soluble calcium which is not dissolved in a neutral solution and is in a particle state, the calcium is suitable for oral administration and is not suitable for injection by an injector, and the product adopts the adhesive which is an anionic polymer and a neutral polymer instead of an adhesive, and the surface of the gastric injury mucosa is also provided with anions, so the product cannot be well adhered to the wound surface of the gastric mucosa, is very easy to fall off, and cannot fully play the role of protecting the gastric mucosa.

Disclosure of Invention

Aiming at the defects in the technical field, the invention provides a brand-new method and a product for protecting the mucosa of the digestive tract, which adopt a physical principle and have strong adhesive force, and the method and the product form a colloid membrane under the acidic environment condition in the digestive tract, thereby playing a role in protecting the mucosa of the digestive tract.

The invention provides a self-curing self-adhesive digestive tract injury mucosa protective gel, which comprises the effective components of gel and biological adhesive; the effective components of the gel are gel forming substances, water-soluble calcium salt and a calcium ion complexing agent; the biological adhesive is polylysine; the pH value of the digestive tract injury mucosa protective gel is not less than 7.

The biological adhesive is polylysine. Polylysine generally has a common molecular weight of 70,000-150,000, 150,000-300,000 and more than 300,000, and the higher the molecular weight is, the stronger the adhesion is, the better the curing effect is, but the relatively complete dissolution is difficult. The invention can comprehensively select the needed polylysine according to the dissolving agent and the like.

Further, the gel-forming substance is one or two of pectin and alginate.

Further, the pectin is one or two of low methoxyl pectin and amidated low methoxyl pectin, and the alginate is one or two of sodium alginate and potassium alginate.

Further, the protective gel also comprises a dissolving agent and an acid-base regulator, wherein the dissolving agent is used for dissolving the gel and the biological adhesive; the pH regulator is used for regulating the pH value of the protective glue to be not less than 7.

Further, the water-soluble calcium is one or more of calcium chloride, calcium lactate and calcium gluconate, and the calcium ion complexing agent is one or two of sodium citrate and EDTA.

Further, the weight ratio of gel to bioadhesive was 3: 1-3; the weight ratio of water-soluble calcium salt to calcium ion complexing agent is 3-6: 0.2-1.5: 1.

the invention also provides a self-curing self-adhesive digestive tract injury mucosa protective adhesive which is independently packaged into two parts, wherein one part is a protective adhesive component, and the other part is a curing liquid; the effective components of the protective gel component are gel and biological adhesive; the effective components of the gel are gel forming substances, water-soluble calcium salt and a calcium ion complexing agent; the biological adhesive is polylysine; the pH value of the digestive tract injury mucosa protective gel is not less than 7; the curing liquid can act with the protective glue to fix the protective film formed by the protective glue at a specified position.

Further, the solidifying liquid is calcium salt solution, and the mass concentration of the calcium salt solution is 0.25-5%.

Further, the gel-forming substance is one or two of pectin and alginate.

The dissolving agent of the present invention is water or a salt acceptable to others, such as a phosphate solution.

The acid-base regulator is sodium hydroxide or potassium hydroxide.

the invention also provides application of the self-curing self-adhesive digestive tract injury mucosa protective gel in preparing a wound medicament for treating peptic ulcer, stress ulcer, gastritis, gastric mucosa resection and dissection, esophageal mucosa dissection and intestinal mucosa dissection.

Since the digestive tract is substantially in an acidic or weakly acidic environment, the object of the present invention can be achieved. The present invention is described primarily in the context of the stomach environment, but is not intended to be limited to the stomach, and is applicable to all digestive tracts of the human body, including the esophagus, stomach, intestine, etc.

Compared with the prior art, the invention has the following advantages:

the invention adopts scientific principle, under the condition of gastric acid, the colloid substance forms colloid membrane on the surface of gastrointestinal mucosa, thereby blocking the digestive action of gastric acid and pepsin on the mucosa and promoting the healing of ulcer.

Before the self-curing adhesive is used, the product is neutral or slightly alkaline in solution state and does not form gel state. Under the acidic condition, the product is formed into a gel film, and the biological adhesive provides the performance of being adhered to gastric mucosa, so that the gel film is adhered to the gastric mucosa, the digestion of gastric acid and pepsin on the mucosa is blocked, and the gastric mucosa is protected.

Based on the ion coagulation mechanism of pectin and alginate as cross-linking matter, calcium ion is complexed by calcium ion complexing agent in alkaline environment and thus can not be cross-linked with the cross-linking matter. After contacting with gastric acid, calcium ions can not be in a complexing state with citric acid ions in an acid environment, and the calcium ions are released, so that pectin or alginate is crosslinked to form a gel film.

The self-curing self-adhesive digestive tract injury mucosa protective gel is prepared by adopting natural high molecular materials extracted from plants, namely sodium alginate and/or pectin and a cross-linking agent, and is sprayed on the surface of the stomach injury mucosa through a gastroscope to form a colloid membrane, so that the stomach mucosa is protected. The product is characterized in that the product is liquid under normal conditions, and under the action of gastric acid, the cross-linking agent in the protective gel is released to form a colloidal membrane on the surface of a gastric injury mucosa, so that the digestive effect of gastric acid and pepsin on gastrointestinal mucosa is avoided, and the healing of ulcer is promoted.

Meanwhile, polylysine used as an adhesive does not react with the gel-forming substance in a slightly alkaline solution and is in a liquid state. The cationic polymer is characterized in the gastric acid environment, and can be crosslinked with a gel-forming substance to enhance the strength of the protective gel, and can be bonded to the wound surface through the cationic property and the anion combination of the gastric mucosa injury surface.

Because the mucosa protective glue can not be contacted with gastric acid immediately after being coated on the damaged mucosa and has the flowing problem, the invention adopts the curing liquid to ensure that the protective glue is quickly fixed on the surface of the damaged mucosa, thereby avoiding the flowing problem of the protective glue.

Detailed Description

The present invention will be described in further detail with reference to examples.

The embodiment of the invention is not particularly described, and the adopted chemicals are all common chemicals which can be purchased in the market. In the examples, polylysine with the molecular weight of 150,000-300,000 is selected by comprehensively considering the dissolving difficulty.

Polylysines of other molecular weights are also possible, as long as dissolution is achieved.

Example 1

1. Potassium alginate 12g

2. Calcium chloride 1 g

3. Sodium citrate 2g

4. Polylysine 10g

Dissolving the above components in 0.01mol phosphate solution (500 ml) adjusted pH to 9.0 with 1mol sodium hydroxide, packaging 5ml per bottle, and sterilizing with high temperature steam. The curing liquid is prepared into 1.1 percent aqueous solution by adopting calcium chloride, each bottle is packed with 5ml, and the high-temperature steam sterilization is carried out.

Example 2

1. Amidated low methoxyl pectin 15 g

2. Calcium chloride 1 g

3. Sodium citrate 2.5 g

4. Polylysine 12g

Dissolving the above components in 0.01mol phosphate solution (pH 9.0 adjusted with 1mol sodium hydroxide) 600ml, packaging each bottle with 5ml, and sterilizing with high temperature steam. The curing liquid is prepared into 1.1 percent aqueous solution by adopting calcium chloride, each bottle is packed with 5ml, and the high-temperature steam sterilization is carried out.

Example 3

1. Sodium alginate 11 g

2. Sodium citrate 4 g

3. Calcium chloride 1.5 g

4. Polylysine 10g

Dissolving the above components in 0.01mol phosphate solution (500 ml) adjusted pH to 9.0 with 1mol sodium hydroxide, packaging 5ml per bottle, and sterilizing with high temperature steam. The curing liquid is prepared into 2 percent aqueous solution by adopting calcium gluconate, each bottle is packed with 5ml, and the high-temperature steam sterilization is carried out.

Example 4

1. Sodium alginate 11 g

2. Sodium citrate 4 g

3. Calcium gluconate 6 g

4. Polylysine 10g

Dissolving the above components in 0.01mol phosphate solution (pH 9.0 adjusted with 1mol sodium hydroxide) 600ml, packaging each bottle with 5ml, and sterilizing with high temperature steam. The curing liquid is prepared into 1.7 percent aqueous solution by adopting calcium chloride, each bottle is packed with 5ml, and the high-temperature steam sterilization is carried out.

Example 5

1. 10g of potassium alginate

2. Sodium citrate 4 g

3. Calcium lactate 2.8 g

4 Polylysine 10g

Dissolving the above components in 0.01mol phosphate solution (pH 9.0 adjusted with 1mol sodium hydroxide) 600ml, packaging each bottle with 5ml, and sterilizing with high temperature steam. The solidified solution is prepared into 5% aqueous solution by adopting calcium lactate, each bottle is packed with 5ml, and the high-temperature steam sterilization is carried out.

Experimental examples evaluation of Effect

The liquids of the above examples were used directly in the experiments.

1 in vitro colloidal film formation test

2ml of the liquid is respectively taken and coated on the inner side surface of fresh pigskin, and then the curing liquid is sprayed to observe the formation of a colloid membrane. As a result, a colloidal film was formed in each of examples 1 to 5.

Test example 2

Safety research test

Examples 1-5 were subjected to the following biological tests

1) Stimulation of oral mucosa: 12ml of the colloidal solution was poured into a sterile petri dish having a diameter of 15cm, then the excess fixative was absorbed by physiological saline 30 minutes after pouring the fixative, and then the test solution was prepared by adding physiological saline at a ratio of 3cm2/ml and leaching at 37 ℃ for 72 hours. The experimental liquid of the sample is made into a cotton ball with the diameter not more than 5mm and soaked in the cheek capsule on one side of 3 golden yellow mice. The contact time is minimum 5min each time, once a day, 4 times totally, after the last contact, the cheek sacs are observed visually 24h, the mice are killed painlessly, tissue samples of representative parts of the cheek sacs are taken and put into 4% formaldehyde solution for fixation, and histological evaluation is carried out after tissue sections are made. The stimulation indexes are all 0, and the tested sample has no oral mucosa irritation.

2) Cytotoxicity: 12ml of the colloidal solution was poured into a sterile petri dish having a diameter of 15cm, then the excess fixative was absorbed by physiological saline 30 minutes after pouring of the fixative, and then the test solution was prepared by adding the culture medium at a ratio of 3cm2/ml and leaching at 37 ℃ for 24 hours. Then, the cytotoxicity is measured by MTT method according to the cytotoxicity test specified in GB/T16886.5, and the cytotoxicity is in the range of 0-1 grade.

3) Sensitization test: 12ml of the colloidal solution was poured into a sterile petri dish having a diameter of 15cm, then the excess fixative was absorbed by physiological saline 30 minutes after pouring the fixative, and then the test solution was prepared by adding physiological saline at a ratio of 3cm2/ml and leaching at 37 ℃ for 72 hours. Then, no sensitization was observed in any of the skin sensitization tests as specified in GB/T16886.10.

Test example 3

Measurement of tissue adhesion (tissue retention method):

examples 1 to 5 were each subjected to the following tests

SD rats are taken and fasted for 24h, are anesthetized by intraperitoneal injection with a sodium pentobarbital solution (40mg/kg), the stomach is dissected and taken out, the stomach is cut out in physiological saline (37 ℃), the inner wall of the stomach is cleaned by the physiological saline, and the cleaned stomach is used within 2 h. Cutting a certain area of stomach tissue (2cm multiplied by 2cm), fixing on a polyethylene film, uniformly coating 0.5ml of protective gel on the wound surface of the stomach mucous membrane, and then spraying the curing liquid. The stomach tissue is placed in a constant humidity closed container with the relative humidity of 92.5 percent for 20 minutes, the treated stomach tissue is fixed on a flushing chute, the angle of the chute is adjusted to 60 ℃, the flow rate of a peristaltic pump is adjusted to 20ml/min, the stomach tissue is flushed with 0.1mol/L hydrochloric acid for 5mins, the flushing liquid is collected in a beaker with a known weight, dried at 70 ℃, weighed, and the tissue adhesion is expressed by the adhesion percentage.

The calculation method is as follows:

percentage of gastric tissue adhesion (Bg/%) Bg/% { [ M- (G-M) ]/M } x 100%

Wherein M is the weight of the mucosa protective glue (0.5ml is dried under the same condition); g is the weight of the empty beaker; g is the total weight of the beaker and the dried residue; m is the amount of solid material contained in the same volume of wash solution (blank control). A larger B value indicates a larger adhesion.

The results show that: example 1: 100%, run 2: 99%, example 3: 100%, example 4: 100%, example 5: 99 percent. Tests show that the bi-component self-adhesive gastric mucosa protective glue of the examples 1 to 5 has strong adhesion to tissues and is not easy to fall off.

Test example 4

Animal testing

Animals were divided into two groups, weighing about 3 kg. The experimental group had 6 animals, and the control group had 6 animals.

Fasting is performed 24 hours before operation, and water is not forbidden.

The anesthesia method comprises the following steps: the rabbit is recommended to be injected with 1.0ml/kg of sodium pentobarbital with the mass concentration of 30 g/L.

The rabbit was fixed on the back on the operating table and the abdomen was dehaired. The test area was disinfected with 2% iodine tincture and 75% ethanol solution as required for routine surgical procedures.

The skin, muscle layer and peritoneum are cut layer by layer in the upper abdomen, and if bleeding is caused, ligation is carried out to stop bleeding. Exposing the stomach, cutting the stomach at the greater curvature, washing the stomach with physiological saline, under the submucosa of the lateral greater curvature of the stomach, 1: 10000 adrenaline normal saline is injected under mucosa, 1ml normal saline is injected to form gastric mucosa protrusion, then a loop device is used for cutting off mucosa with the diameter of 1cm to form a wound surface, thrombin (1ml contains 50U of thrombin) is sprayed and hemostasis is completely pressed, and the diameter of the wound surface is measured. The control group was not treated, and the test group was coated with 0.5ml of a protective gel, sprayed with a curing liquid to form a solidified protective film, and then sutured to the stomach. Then the abdominal wall is sutured layer by layer. Placing in a feeding cage, and fasting for one day.

And (4) observing results: euthanizing the animals 1 week after the operation, cutting the stomach wall along the original incision, observing the healing condition of the wound surface, measuring the diameter of the wound surface, and finding that 6 rabbits in the experimental group with ulcer 1 week after the operation heal with the healing rate of 100 percent; the control group had 2 healed, 4 not healed, and the healing rate was 33%. The above results show that the healing of the ulcer due to gastric mucosal injury can be significantly promoted and the therapeutic effect on gastric ulcer can be significantly enhanced according to example 1.

The specific embodiments described herein are merely illustrative of the spirit of the invention. Various modifications or additions may be made to the described embodiments or alternatives may be employed by those skilled in the art without departing from the spirit of the invention.

Claims

1. The self-curing self-adhesive digestive tract injury mucosa protective gel is characterized in that the effective components of the digestive tract injury mucosa protective gel are gel and biological adhesive; the effective components of the gel are gel forming substances, water-soluble calcium salt and a calcium ion complexing agent; the biological adhesive is polylysine; the pH value of the digestive tract injury mucosa protective gel is not less than 7.

2. The self-curing self-adhesive mucosa protective gel for digestive tract injury according to claim 1, wherein the gel-forming substance is one or two of pectin and alginate.

3. The self-curing self-adhesive mucosa protective gel for digestive tract injury according to claim 2, wherein the pectin is one or two of low methoxyl pectin and amidated low methoxyl pectin, and the alginate is one or two of sodium alginate and potassium alginate.

4. The self-curing self-adhesive digestive tract injury mucosa protective gel according to claim 1, further comprising a dissolving agent and an acid-base regulator, wherein the dissolving agent is used for dissolving gel and biological adhesive; the pH regulator is used for regulating the pH value of the protective glue to be not less than 7.

5. The self-curing self-adhesive mucosa protective gel for digestive tract injury according to claim 1, wherein the water-soluble calcium is one or more of calcium chloride, calcium lactate and calcium gluconate, and the calcium ion complexing agent is one or two of sodium citrate and EDTA.

6. The self-curing self-adhesive digestive tract injury mucosa protective gel according to claim 1, wherein the weight ratio of the gel to the biological adhesive is 3: 1-3; the weight ratio of water-soluble calcium salt to calcium ion complexing agent is 3-6: 0.2-1.5: 1.

7. a self-curing self-adhesive digestive tract injury mucosa protective gel is characterized in that the digestive tract injury mucosa protective gel comprises two parts which are independently packaged, wherein one part is a protective gel component, and the other part is a curing liquid; the effective components of the protective gel component are gel and biological adhesive; the effective components of the gel are gel forming substances, water-soluble calcium salt and a calcium ion complexing agent; the biological adhesive is polylysine; the pH value of the digestive tract injury mucosa protective gel is not less than 7; the curing liquid can act with the protective glue to fix the protective film formed by the protective glue at a specified position.

8. The self-curing self-adhesive digestive tract injury mucosa protective gel according to claim 7, wherein the curing liquid is calcium salt solution, and the mass concentration of the calcium salt solution is 0.25-5%.

9. The self-curing self-adhesive mucosa protective gel for digestive tract injury according to claim 7, wherein the gel-forming substance is one or two of pectin and alginate.

10. The self-curing self-adhesive mucosa protective gel for digestive tract injury according to claim 8, wherein the pectin is one or two of low methoxyl pectin and amidated low methoxyl pectin, and the alginate is one or two of sodium alginate and potassium alginate.

11. The self-curing self-adhesive digestive tract injury mucosa protective gel according to claim 7, further comprising a dissolving agent and an acid-base regulator, wherein the dissolving agent is used for dissolving gel and biological adhesive; the pH regulator is used for regulating the pH value of the protective glue to be not less than 7.

12. The self-curing self-adhesive mucosa protective gel for digestive tract injury according to claim 7, wherein the water-soluble calcium is one or more of calcium chloride, calcium lactate and calcium gluconate, and the calcium ion complexing agent is one or two of sodium citrate and EDTA.

13. The self-curing self-adhesive digestive tract injury mucosa protective gel according to claim 7, wherein the weight ratio of the gel to the biological adhesive is 3: 1-3; the weight ratio of water-soluble calcium salt to calcium ion complexing agent is 3-6: 0.2-1.5: 1.

14. use of the self-curing adhesive mucosa protectant of digestive tract injury according to any of claims 1 or 7 for the treatment of wounds after peptic ulcer, stress ulcer, gastritis, gastric mucosal resection and dissection, esophageal mucosa dissection and intestinal mucosa dissection.