WO2020182139A1 - 消化道黏膜保护胶 - Google Patents

消化道黏膜保护胶 Download PDF

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Publication number
WO2020182139A1
WO2020182139A1 PCT/CN2020/078733 CN2020078733W WO2020182139A1 WO 2020182139 A1 WO2020182139 A1 WO 2020182139A1 CN 2020078733 W CN2020078733 W CN 2020078733W WO 2020182139 A1 WO2020182139 A1 WO 2020182139A1
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Prior art keywords
protective glue
gastrointestinal
gastrointestinal mucosa
gel
mucosal
Prior art date
Application number
PCT/CN2020/078733
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English (en)
French (fr)
Inventor
戴建英
Original Assignee
杭州英健生物科技有限公司
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from CN201910184773.2A external-priority patent/CN109758580A/zh
Priority claimed from CN201910255736.6A external-priority patent/CN110075345B/zh
Priority claimed from CN201910448532.4A external-priority patent/CN112076341A/zh
Priority claimed from CN201910496774.0A external-priority patent/CN112138202A/zh
Priority claimed from CN201910977789.9A external-priority patent/CN110585489A/zh
Application filed by 杭州英健生物科技有限公司 filed Critical 杭州英健生物科技有限公司
Priority to JP2021554415A priority Critical patent/JP7325853B2/ja
Priority to KR1020217029900A priority patent/KR20210131375A/ko
Publication of WO2020182139A1 publication Critical patent/WO2020182139A1/zh

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L24/00—Surgical adhesives or cements; Adhesives for colostomy devices
    • A61L24/04—Surgical adhesives or cements; Adhesives for colostomy devices containing macromolecular materials
    • A61L24/08—Polysaccharides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L24/00—Surgical adhesives or cements; Adhesives for colostomy devices
    • A61L24/001—Use of materials characterised by their function or physical properties
    • A61L24/0015—Medicaments; Biocides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L24/00—Surgical adhesives or cements; Adhesives for colostomy devices
    • A61L24/001—Use of materials characterised by their function or physical properties
    • A61L24/0031—Hydrogels or hydrocolloids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L24/00—Surgical adhesives or cements; Adhesives for colostomy devices
    • A61L24/04—Surgical adhesives or cements; Adhesives for colostomy devices containing macromolecular materials
    • A61L24/10—Polypeptides; Proteins
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
    • A61L31/04—Macromolecular materials
    • A61L31/042—Polysaccharides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
    • A61L31/04—Macromolecular materials
    • A61L31/043—Proteins; Polypeptides; Degradation products thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
    • A61L31/14—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
    • A61L31/148—Materials at least partially resorbable by the body
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
    • A61L31/14—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
    • A61L31/16—Biologically active materials, e.g. therapeutic substances
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/04—Drugs for disorders of the alimentary tract or the digestive system for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
    • A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
    • A61L2300/404—Biocides, antimicrobial agents, antiseptic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2400/00—Materials characterised by their function or physical properties
    • A61L2400/04—Materials for stopping bleeding
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2400/00—Materials characterised by their function or physical properties
    • A61L2400/06—Flowable or injectable implant compositions

Definitions

  • the invention relates to the technical field of biomedicine, in particular to a protective glue for the mucosa of the digestive tract.
  • Endoscopic gastric mucosal resection and gastric mucosal dissection are minimally invasive surgery for the removal of early gastric cancer and benign gastric tumors. It can not only achieve the purpose of radical treatment of early gastric cancer, but also has the advantages of less trauma and less impact on the quality of life of patients. Gradually replace some traditional surgical procedures. Minimally invasive surgery for early canceration of the esophagus and colon and removal of benign tumors has also replaced some traditional surgical operations. However, at present, only hemostatic treatment is performed on the wound after this operation, and there is no good wound protection measure. Bare wounds experience active ulcer, healing and scarring periods under the action of digestive juice, and the wound healing time is longer.
  • Cida Patent discloses a gastric mucosal protective glue, which includes a gel-forming substance, an acid-base regulator, a cross-linking agent and an adhesive; the weight ratio of the gel-forming substance, the cross-linking agent and the adhesive It is 1:0.01-0.3:0.1-0.5, and the amount of acid-base regulator is adjusted to pH 6.5-8.5.
  • the patent only uses pure physical principles to protect the gastric mucosa.
  • the crosslinking agent used is acid-soluble calcium, which is insoluble in a neutral solution and is in a granular state.
  • the adhesive used in this product is an anionic polymer and a neutral polymer instead of an adhesive.
  • the gastric mucosal surface is also anionic, so it cannot adhere to the gastric mucosal wound surface very well, and it is very easy to fall off. Give full play to the protective role of the gastric mucosa.
  • the present invention provides a new method and product for protecting the mucosa of the digestive tract, which adopts physical principles and has strong adhesion, and plays a role in protecting the mucosa of the digestive tract.
  • the present invention provides a protective glue for the mucosa of the digestive tract, comprising a protective layer and a biological adhesive; wherein the effective ingredients of the protective layer are gel-forming substances, water-soluble calcium salts and calcium ion complexing agents; the gel-forming substances are pectin and seaweed One or more of acid salt, deacetylated gellan gum; biological adhesives are carboxymethyl cellulose, carboxymethyl chitosan, carbomer, ethyl cellulose, hydroxypropyl cellulose, hydroxy One or more of propyl methyl cellulose, Bletilla striata polysaccharide, polylysine, mussel viscous protein, collagen, polydopamine, sodium hyaluronate, and chitosan quaternary ammonium salt; the digestive tract
  • the pH value of mucosal protective glue is not less than 7.
  • the calcium ion complexing agent is one or more of citric acid, citrate, EDTA or EDTA sodium salt.
  • the pectin is one or two of low methoxy pectin and amidated low methoxy pectin.
  • the alginate is one or two of sodium alginate or potassium alginate.
  • the mass concentration of calcium salt in the calcium salt solution is 0.25% to 5%.
  • the weight ratio of the gel-forming substance to the calcium ion complexing agent is 3-6:1.
  • the bioadhesive is polylysine, and the weight ratio of the gel-forming substance to the polylysine is 3:1-3.
  • Polylysine generally has a molecular weight of 70,000 to 150,000, 150,000 to 300,000 and >300,000. The larger the molecular weight, the stronger the adhesion and the better curing effect, but it is relatively completely dissolved. difficult.
  • the required polylysine can be selected comprehensively according to the dissolving agent and the like.
  • the weight ratio of the gel-forming substance, the water-soluble calcium salt, and the calcium ion complexing agent is 3-6:0.2-1.5:1.
  • the protective glue also includes a dissolving agent and an acid-base regulator.
  • the dissolving agent is used to dissolve the gel and biological adhesive; the acid-base regulator is used to adjust the pH of the protective glue to not less than 7.
  • the dissolving agent of the present invention is water or other salts acceptable to the human body, such as a phosphate solution.
  • the acid-base regulator is sodium hydroxide and potassium hydroxide.
  • the biological adhesive is an aqueous solution containing mussel mucin. Utilizing the self-adhesive properties of the mussel protein-containing adhesive layer, the gastric mucosa and the gastric mucosa protective glue are tightly connected.
  • the weight ratio of the gel-forming substance, water-soluble calcium and calcium complexing agent is 3-6:0.2-1.5:1, and the total weight of the gel-forming substance, water-soluble calcium and calcium complexing agent accounts for the protective layer group 2.5-3.5wt% of points.
  • the gel forming components are pectin, alginate, water-soluble calcium and calcium complexing agent.
  • the weight ratio of pectin, alginate, water-soluble calcium and calcium complexing agent is 2-3:1-2:0.2-1.5:1.
  • mussel mucin is 5-25 wt% of the bioadhesive.
  • the protective layer component also contains 0.3-5 wt% of a plasticizer.
  • the plasticizer is one or more of glycerin, polyethylene glycol, polyvinyl alcohol, mannitol and sorbitol.
  • the pH adjusting agent of the adhesive layer component is one or more of acetic acid, hydrochloric acid, carbonic acid, citric acid, malic acid, gluconic acid, lactic acid, oxalic acid, ascorbic acid, tartaric acid and benzoic acid.
  • the protective layer group The pH adjusters of the sub-components are sodium bicarbonate, sodium carbonate, potassium carbonate, dipotassium hydrogen phosphate, disodium hydrogen phosphate, trisodium phosphate, tripotassium phosphate, sodium citrate, potassium citrate, sodium hydroxide, potassium hydroxide and One or more of monosodium citrate.
  • the gastrointestinal mucosa protective glue further includes a temperature-sensitive biological material and a gelation temperature regulator.
  • the temperature-sensitive biological material the gelation temperature regulator: the bioadhesive agent: the gel-forming substance has a weight percentage of 1-30:0.5-5:0.3-5:0.2-1.
  • the temperature-sensitive biological material is poly-N-isopropylacrylamide and its derivatives, hydroxybutyl chitosan and its derivatives, poloxamer 407, polylactic acid/glycolic acid/polyethylene glycol Copolymer, polyethylene glycol monomethyl ether-(sebacic acid-D,L-lactic acid) polyester anhydride-polyethylene glycol monomethyl ether triblock copolymer, cellulose and its derivatives, and polyethylene glycol /One or more of polycaprolactone block copolymers.
  • the gelation temperature regulator is one or more of poloxamer 188, polysorbate, and polyethylene glycol.
  • the components of the colloidal liquid are dissolved in the dissolving liquid to form a colloidal solution; the phase transition temperature of the protective glue is 25°C-40°C.
  • the dissolving solution is one or more of pure water, physiological saline and phosphate solution.
  • the protective glue in the present invention also includes a fixing solution, which is a water-soluble calcium solution.
  • water-soluble calcium is one or more of calcium chloride, calcium lactate, and calcium gluconate.
  • the concentration of the water-soluble calcium solution is 0.3%-5%.
  • the protective glue also includes a pharmaceutical compound or composition with an effective drug amount for treating ulcers and tumors.
  • the invention also provides a protective glue that also includes a gastric acid inhibitor and an anti- Helicobacter preparation.
  • gastric acid inhibitors are proton pump inhibitors or H 2 receptor antagonists.
  • a pH regulator needs to be added.
  • the anti- Helicobacter preparation is composed of one or more of bismuth, metronidazole or tinidazole, amoxicillin, clarithromycin, tetracycline, and furazolidone.
  • the weight ratio of the gastric acid inhibitor, the anti- Helicobacter preparation and the other components of the protective gel is 0.001-0.05:0.001-1:1-20.
  • the gastrointestinal mucosal protective glue also includes ingredients that promote the proliferation and migration of gastrointestinal mucosal epithelial cells, including glycyrrhizin, ⁇ -glucan, codonopsis polysaccharide, sola gum, pleurotus eryngii glucan, oat glucan, One or more of Grifola frondosa polysaccharide, grain glucan, yeast glucan, and Ganoderma lucidum polysaccharide.
  • the weight ratio of these components in the entire protective glue is 0.001%-0.5%.
  • the present invention also provides a pharmaceutical composition containing the aforementioned gastrointestinal mucosal protective glue and other drugs with therapeutic effects.
  • the drugs with therapeutic effects referred to in the present invention can be various drugs for treating digestive tract diseases.
  • the present invention also provides an injection for uplift of the digestive tract, comprising the aforementioned protective glue for the mucosa of the digestive tract.
  • the submucosal injection protruding agent of the digestive tract is injected below the mucosa to separate the mucosal layer from the muscle layer and facilitate mucosal peeling.
  • the present invention also provides the aforementioned gastrointestinal mucosal protective glue in the vegetation treatment of peptic ulcer, stress ulcer, ulcerative colitis, gastritis, gastric mucosal resection and stripping wounds, esophageal mucosal dissection and intestinal mucosal dissection Application in wounds, gastrinoma and gastric cancer drugs.
  • the digestive tract is basically in an acidic or weakly acidic environment, the objectives of the present invention can be achieved.
  • the description is mainly based on the environment of the stomach, but it does not mean that the present invention is limited to the stomach.
  • the present invention can be applied to all digestive tracts of the human body, including the esophagus, stomach, and intestines.
  • the present invention has the following advantages:
  • the invention adopts scientific principles to make the gelatinous substance form a colloidal film on the gastrointestinal mucosa surface under the condition of gastric acid, block the digestive effect of gastric acid and pepsin on the mucosa, and promote ulcer healing.
  • the gastrointestinal mucosal protective glue of the present invention is a self-curing and self-adhesive gastrointestinal mucosal protective glue product. Before use, it is neutral or slightly alkaline in a solution state, and the product does not form a gel state. Under acidic conditions, the product is made into a gel film, and the bioadhesive provides the performance of adhering to the gastric mucosa, allowing the gel film to adhere to the gastric mucosa, blocking the digestion of gastric acid and pepsin on the mucosa. Play the role of protecting the gastric mucosa.
  • the present invention is based on the ion coagulation mechanism of transaction material pectin and alginate.
  • calcium ion is complexed by calcium ion complexing agent, so it cannot be crosslinked with colloid-forming material.
  • calcium ions After being in contact with gastric acid, calcium ions cannot be in a complex state with citrate ions in an acidic environment, and calcium ions are released to crosslink pectin or alginate to form a gel film.
  • the protective glue is made of natural polymer materials extracted from plants, sodium alginate and/or pectin, and a cross-linking agent. It is sprayed on the gastric mucosal surface through a gastroscope to form a colloidal film to protect the gastric mucosa.
  • the characteristic of this product is that it is liquid under normal circumstances. Under the action of gastric acid, the cross-linking agent in the protective glue is released, forming a colloidal film on the surface of the gastric mucosa, avoiding the digestion effect of gastric acid and pepsin on the gastrointestinal mucosa. Promote the healing of ulcers.
  • the adhesive used at the same time does not interact with the gel-forming substance in the alkaline solution and is in a liquid state.
  • it exhibits the characteristics of cationic polymer.
  • One is that it is cross-linked with colloidal substances to enhance the strength of the protective glue.
  • the other is that it can also bind to the anion on the surface of the gastric mucosa through its cationic properties. Wound surface.
  • the curing liquid is used in the present invention to quickly fix the protective glue on the damaged mucosal surface, avoiding the flow problem of the protective glue.
  • the present invention provides a self-curing two-component ion and temperature dual-sensitive gastrointestinal mucosal protective glue with dual ion and temperature sensitivity.
  • Self-fixing two-component temperature-sensitive gastrointestinal mucosal protective glue includes colloidal fluid and fixative.
  • the colloidal fluid includes temperature-sensitive materials, gelation temperature regulator, bioadhesive and ion-sensitive fixative.
  • the fixative is water-soluble Calcium solution.
  • the colloidal fluid is liquid at a temperature below room temperature (25°C).
  • the spray fixative fluid is quickly fixed, while at body temperature (37°C), temperature-sensitive materials are
  • body temperature 37°C
  • temperature-sensitive materials are The phase change occurs at body temperature, which makes the product change from liquid to gel through phase change, and the adhesive provides the performance of bonding to the damaged mucosa, making the gel adhere to the mucosa and not easy to fall off, blocking the digestion of the mucosa by the digestive juice It plays a role in protecting the mucous membrane from damage.
  • the calcium ions in the curing liquid and the ion-sensitive fixative in the colloidal liquid form a gel film to avoid the loss of the colloidal liquid and strengthen the protection of the damaged mucosa.
  • the binder is polylysine.
  • the polylysine with a molecular weight of 150,000 to 300,000 which is selected in the embodiments considering the difficulty of dissolution, has a molecular weight of 150,000 to 300,000.
  • Polylysines of other molecular weights are also possible, as long as they are dissolved.
  • the above ingredients are dissolved in 500ml of 0.01mol of phosphate solution adjusted by 1mol of sodium hydroxide to adjust pH9.0, 5ml per bottle, and high temperature steam sterilization.
  • the solidification liquid is made into 1.1% aqueous solution with calcium chloride, packaged in 5ml per bottle, and sterilized by high temperature steam.
  • the above ingredients are dissolved in 600ml of 0.01mol of phosphate solution adjusted by 1mol of sodium hydroxide to adjust pH9.0. Each bottle is packed with 5ml and sterilized by high temperature steam. The solidification liquid is made into 1.1% aqueous solution with calcium chloride, packaged in 5ml per bottle, and sterilized by high temperature steam.
  • the above ingredients are dissolved in 500ml of 0.01mol of phosphate solution adjusted by 1mol of sodium hydroxide to adjust pH9.0, 5ml per bottle, and high temperature steam sterilization.
  • the solidifying liquid is made of calcium gluconate into a 2% aqueous solution, packaged in 5ml per bottle, and sterilized by high-temperature steam.
  • the above ingredients are dissolved in 600ml of 0.01mol pH9.0 phosphate solution adjusted by 1mol sodium hydroxide, 5ml per bottle, and high temperature steam sterilization.
  • the solidification liquid is prepared with calcium chloride into a 1.7% aqueous solution, packaged in 5ml per bottle, and sterilized by high-temperature steam.
  • the above ingredients are dissolved in 600ml of 0.01mol pH9.0 phosphate solution adjusted by 1mol sodium hydroxide, 5ml per bottle, and high temperature steam sterilization.
  • the solidification liquid is prepared with calcium lactate into a 5% aqueous solution, packaged in 5ml per bottle, and sterilized by high-temperature steam.
  • Example 1 to Example 5 all formed colloidal films.
  • Oral mucosal irritation Pour 12ml of colloidal fluid into a sterile petri dish with a diameter of 15cm, and then pour it into the fixative for 30 minutes, rinse with saline to absorb the excess fixative, and then add it to the physiological at the ratio of 3cm 2 /ml
  • the brine was leached at 37 degrees Celsius for 72 hours to prepare the test solution.
  • the test solution of the sample was made into cotton balls with a diameter of no more than 5 mm, soaked and placed in the cheek pouch of 3 golden hamsters. The contact time is at least 5 minutes each time, once a day, 4 times in total. 24 hours after the last contact, the cheek pouch was observed visually, and the hamsters were sacrificed without pain.
  • the tissue samples of the representative part of the cheek pouch were taken and fixed in 4% formaldehyde solution to make the tissue. Histological evaluation was performed after sectioning. Results The irritation index was all 0, and the tested samples had no oral mucosa irritation.
  • Cytotoxicity Pour 12ml of colloidal fluid into a sterile petri dish with a diameter of 15cm, then pour it into the fixative solution 30 minutes later, wash with saline to absorb the excess fixative solution, and then add the culture medium at a ratio of 3cm 2 /ml , Leaching at 37 degrees Celsius for 24 hours to prepare the test solution. Then, according to the cytotoxicity test specified in GB/T16886.5, the cytotoxicity is determined by the MTT method, and the cytotoxicity is in the range of 0-1.
  • Sensitization test Pour 12ml of colloidal fluid into a sterile petri dish with a diameter of 15cm, and then pour it into the fixative for 30 minutes, rinse with saline to absorb the excess fixative, and then add it to the body at a ratio of 3cm 2 /ml The brine was leached at 37 degrees Celsius for 72 hours to prepare the test solution. Then the skin sensitization test was carried out according to the method specified in GB/T16886.10, and no sensitization was observed.
  • tissue adhesion tissue retention method
  • the calculation method is as follows:
  • Bg/% ⁇ [M-(G-g-m)]/M ⁇ x100%
  • M is the weight of the mucosal protective glue (0.5ml dried under the same conditions); g is the weight of the empty beaker; G is the total weight of the beaker and the residue after drying; m is the amount of solid matter in the same volume of rinse solution (blank Control). The greater the B value, the greater the adhesion.
  • Example 1 100%
  • Implementation 2 99%
  • Example 3 100%
  • Example 4 100%
  • Example 5 99%
  • the test shows that the protective glue for the digestive tract mucosa of Examples 1-5 has strong adhesion to the tissue and is not easy to fall off.
  • the animals were divided into two groups, weighing about 3kg. There were 6 in the experimental group and 6 in the control group.
  • Anesthesia It is recommended to inject 1.0ml/kg of sodium pentobarbital with a mass concentration of 30g/L in rabbits.
  • the rabbit is fixed on the operating table on its back, and the abdomen is hairless.
  • the test area was disinfected with 2% iodine tincture and 75% ethanol solution according to the requirements of routine surgery.
  • the control group did not do any treatment.
  • the experimental group was coated with 0.5ml protective glue, and then sprayed with the curing liquid to form a solidified protective film, and then the stomach was sutured. Then suture the abdominal wall layer by layer. Put it in a breeding cage and fast for one day.
  • the binder is polylysine.
  • Adhesive layer components are:
  • the above ingredients are dissolved in distilled water, the pH is adjusted to 8.5 by sodium hydroxide, and the solution is prepared with a mass concentration of 3.0%, and then 1g of glycerin is added per 100ml of the solution, packed in 5ml per bottle, and sterilized by high-temperature steam.
  • the above ingredients are dissolved in distilled water, the pH is adjusted to 8.5 by sodium hydroxide, the mass concentration is 2.5%, and 5g of polyvinyl alcohol is added per 100ml, packaged with 5ml per bottle, and stored under high temperature steam sterilization.
  • Wounds of gastric mucosa are those after peptic ulcer, stress ulcer, gastritis, gastric mucosal resection and stripping.
  • the wound depth is based on 0.5cm
  • the adhesive layer components are sprayed at 0.2-0.25 times the wound area, that is, 0.2-0.25mL of adhesive layer components are sprayed on the wound surface of 1cm2.
  • the protective layer component is sprayed in an amount of 0.3-0.5 times the wound area, that is, 0.3-0.5 mL of the adhesive layer component is sprayed on the wound surface of 1 cm 2 .
  • the amount of adhesive layer components and protective layer components shall be adjusted according to the ratio of the actual wound depth to the reference depth.
  • the adhesive layer components of the self-adhesive gastric mucosa protective glue are sprayed onto the wound surface to form an adhesive layer, and the protective layer components are sprayed onto the adhesive layer to form a protective layer.
  • the first interface the interface between the protective layer and the adhesive layer, combined with the anion of alginic acid through the cationic properties of mussel mucin.
  • the second interface the interface between the adhesive layer and the wound of the gastric mucosa, through the cationic properties of mussel mucin and the anion of the wound tissue cells, and under physiological conditions part of the hydrophobic groups of mussel mucin are exposed to the lipids of the cell membrane
  • the qualitative bilayers bind to each other through hydrophobic interaction.
  • Adhesive layer adhesive layer components in the mussels mucin Duo Baji some phenolic hydroxyl groups oxidized to quinones, oxidation of DOPA and DOPA unoxidized form a high molecular crosslinked polymer network.
  • Protective layer Under the action of gastric acid, a protective film is formed through ionic crosslinking and curing.
  • the components of the adhesive layer are first applied to the damaged mucosal surface, that is, the wound surface to form an adhesive layer. Because the mussel mucin in the adhesive layer has the characteristics of positively charged and negatively charged cells on the wound surface firmly bound, mussel adhesive layer additionally several amino acid mucin-containing hydrophobic groups, in the presence of physiological solution, mussel adhesive protein hydrophobic groups exposed portion, but also through the cell membrane lipid bilayer The hydrophobic interactions combine with each other.
  • the positive charge of the adhesive layer can also be combined with the negatively charged alginate in the protective layer through ionic bonds. Since the protective layer is alkaline, when the pH is increased by diffusion into the adhesive layer, some of the phenolic hydroxyl groups in the mussel mucin dopa group in the adhesive layer are oxidized to quinones. This process in the body catechol It will be accelerated presence of the oxidase, oxidized and unoxidized dopa dopa crosslinking the adhesive layer to form a polymer network polymer.
  • the acidity of the adhesive layer will also diffuse to the protective layer, causing the protective layer to internally cross-link, so that the damaged gastric mucosa and the protective layer can be tightly connected, so that the protective layer can be held firmly on the wound surface.
  • the protective layer forms a protective film through ionic cross-linking and solidification under the action of gastric acid.
  • the added plasticizer makes the protective film have a certain degree of elasticity and resistance to gastric peristalsis.
  • Intradermal irritation 0.2 ml of the test solution of the sample was injected into the skin, and the edema and redness were observed 2 hours, 24 hours and 72 hours after injection. The reaction of all the examples was 0-1.
  • Cytotoxicity The samples are determined by MTT method according to the cytotoxicity test specified in GB/T16886.5, and the cytotoxicity is in the range of 0-1.
  • tissue adhesion tissue retention method
  • the calculation method is as follows:
  • Bg/% ⁇ [M-(G-g-m)]/M ⁇ x100%
  • M is the weight of the mucosal protective glue (0.5ml dried under the same conditions); g is the weight of the empty beaker; G is the total weight of the beaker and the residue after drying; m is the amount of solid matter in the same volume of rinse solution (blank Control). The greater the B value, the greater the adhesion.
  • Example 1 98%
  • Example 2 99%
  • Example 3 97%
  • Example 4 95%
  • Example 5 99%.
  • Tests show that the protective glue of the digestive tract mucosa of Examples 6-10 has strong adhesion to the tissue and is not easy to fall off.
  • the animals were divided into two groups, weighing about 3kg. There were 6 in the experimental group and 6 in the control group.
  • Anesthesia It is recommended to inject 1.0ml/kg of sodium pentobarbital with a mass concentration of 30g/L in rabbits.
  • the rabbit is fixed on the operating table on its back, and the abdomen is hairless.
  • the test area was disinfected with 2% iodine tincture and 75% ethanol solution according to the requirements of routine surgery.
  • the control group did not do any treatment.
  • the experimental group was first coated with 0.2ml of the adhesive layer component, and then coated with 0.4ml of the protective layer component. It can be seen that the protective layer component formed a solidified protective film after contacting gastric acid, and then the stomach was sutured. . Then suture the abdominal wall layer by layer. Put it in a breeding cage and fast for one day.
  • the adhesive is polylysine, and temperature sensitive materials are added at the same time.
  • the above ingredients are dissolved in 60ml pure water at 4°C, completely dissolved into a transparent liquid, and then the volume is fixed to 100ml, and each bottle is packed in 5ml. High temperature steam sterilization. Store at low temperature.
  • the solidification liquid is made into 1.1% aqueous solution with calcium chloride, packaged in 5ml per bottle, and sterilized by high temperature steam.
  • the above ingredients are dissolved in 60ml of pure water at 4°C, completely dissolved into a transparent liquid, and then the volume is fixed to 100ml, each bottle is 5ml packaging, and high temperature steam sterilization. Store at low temperature.
  • the solidification liquid is made into 1.1% aqueous solution with calcium chloride, packaged in 5ml per bottle, and sterilized by high temperature steam.
  • the above ingredients are dissolved in 60ml of pure water at 4°C, completely dissolved into a transparent liquid, and then the volume is fixed to 100ml, each bottle is 5ml packaging, and high temperature steam sterilization. Store at low temperature.
  • the solidifying liquid is made of calcium gluconate into a 2% aqueous solution, packaged in 5ml per bottle, and sterilized by high-temperature steam.
  • the above ingredients are dissolved in 60ml of pure water at 4°C, completely dissolved into a transparent liquid, and then the volume is fixed to 100ml, each bottle is 5ml packaging, and high temperature steam sterilization. Store at low temperature.
  • the solidification liquid is prepared with calcium chloride into a 1.7% aqueous solution, packaged in 5ml per bottle, and sterilized by high-temperature steam.
  • the solidification liquid is prepared with calcium lactate into a 5% aqueous solution, packaged in 5ml per bottle, and sterilized by high-temperature steam.
  • Example 11 to Example 15 all formed colloidal films.
  • Oral mucosa irritation Pour 5ml of colloidal fluid into a sterile petri dish with a diameter of 9cm, spray the fixative, and place it in a 37°C incubator for 30 minutes to form a film, and then add physiological saline at a ratio of 3cm 2 /ml , The test solution was prepared by leaching at 37°C for 72 hours.
  • the test solution of the sample was made into cotton balls with a diameter of no more than 5 mm, soaked and placed in the cheek pouch of 3 golden hamsters.
  • the contact time is at least 5 minutes each time, once a day, 4 times in total. 24 hours after the last contact, the cheek pouch was observed visually, and the hamsters were sacrificed without pain.
  • the tissue samples of the representative part of the cheek pouch were taken and fixed in 4% formaldehyde solution to make the tissue. Histological evaluation was performed after sectioning. Results The irritation index was all 0, and the tested samples had no oral mucosa irritation.
  • Cytotoxicity Pour 5ml of colloidal liquid into a sterile petri dish with a diameter of 9cm, spray the fixative solution, and place it in a 37°C incubator for 30 minutes to form a film, and then add the culture medium at a ratio of 3cm 2 /ml.
  • the test solution was prepared by leaching at 37°C for 24 hours. Then, according to the cytotoxicity test specified in GB/T16886.5, the MTT method was used to determine the cell proliferation rate in the range of 76%-89%.
  • Sensitization test Pour 5ml of colloidal fluid into a sterile petri dish with a diameter of 9cm, spray the fixative, and place it in a 37°C incubator for 30 minutes to make it into a film, and then add it to the body at a ratio of 3cm 2 /ml The brine was leached at 37°C for 72 hours to prepare a test solution. Then the skin sensitization test was carried out according to the method specified in GB/T16886.10, and no sensitization was observed.
  • tissue adhesion tissue retention method
  • the calculation method is as follows:
  • Bg/% ⁇ [M-(G-g-m)]/M ⁇ x100%
  • M is the weight of the mucosal protective glue (0.5ml dried under the same conditions); g is the weight of the empty beaker; G is the total weight of the beaker and the residue after drying; m is the amount of solid matter in the same volume of rinse solution (blank Control). The greater the B value, the greater the adhesion.
  • Example 11 90%, Example 12: 91%, Example 13: 91%, Example 14: 92%, and Example 15: 90%.
  • Tests show that the temperature-sensitive mucosal protective glues of Examples 11-15 have strong adhesion to tissues and are not easy to fall off.
  • the animals were divided into two groups, weighing about 3kg. There were 6 in the experimental group and 6 in the control group.
  • Anesthesia It is recommended to inject 1.0ml/kg of sodium pentobarbital with a mass concentration of 30g/L in rabbits.
  • the rabbit is fixed on the operating table on its back, and the abdomen is hairless.
  • the test area was disinfected with 2% iodine tincture and 75% ethanol solution according to the requirements of routine surgery.
  • the above ingredients are dissolved in 1500ml of physiological salt solution adjusted to pH8.0 with 1mol sodium hydroxide, packaged in 15ml per bottle, and sterilized by high temperature steam to obtain an injection protruding agent.
  • the injection protruding agent prepared in Examples 16-20 was subjected to the following biological tests
  • the test solution was prepared by adding physiological saline at a ratio of 0.2g/ml.
  • the test solution of the sample was made into cotton balls with a diameter of no more than 5 mm, soaked in the cheek pouch of 3 golden hamsters.
  • the contact time is at least 5 minutes each time, once a day, 4 times in total. 24 hours after the last contact, the cheek pouch was observed visually, and the hamsters were sacrificed without pain.
  • the tissue samples of the representative part of the cheek pouch were taken and fixed in 4% formaldehyde solution to make the tissue. Histological evaluation was performed after sectioning. Results The irritation index was all 0, and the tested samples had no oral mucosa irritation.
  • Cytotoxicity The test solution was prepared by adding the culture medium at the ratio of 0.2g/ml. Then, according to the cytotoxicity test specified in GB/T16886.5, the cytotoxicity is determined by the MTT method, and the cytotoxicity is in the range of 0-1.
  • Sensitization test The test solution was prepared by adding physiological saline at a ratio of 0.2g/ml. Then the skin sensitization test was carried out according to the method specified in GB/T16886.10, and no sensitization was observed.
  • the above ingredients are dissolved in distilled water, configured to a mass concentration of 3%, 2.5ml per bottle, and high temperature steam sterilization.
  • the above ingredients are dissolved in distilled water, configured to a mass concentration of 3%, 2.5ml per bottle, and sterilized.
  • Baking soda (pH adjuster): Adjust the pH to 8.0.
  • the proton pump inhibitor lyophilized powder is dissolved in a special solvent. During the configuration process, keep the pH weakly alkaline.
  • the above ingredients are dissolved in distilled water, configured to a mass concentration of 3%, 2.5ml per bottle, and sterilized.
  • the raw materials of the pharmaceutical composition are as follows:
  • Baking soda (pH adjuster): Adjust the pH to 8.0.
  • the proton pump inhibitor lyophilized powder is dissolved in a special solvent. During the configuration process, keep the pH weakly alkaline.
  • the above ingredients are dissolved in distilled water, configured to a mass concentration of 3%, 2.5ml per bottle, and sterilized.
  • the raw materials of the pharmaceutical composition are as follows:
  • Baking soda (pH adjuster): Adjust the pH to 8.0.
  • the proton pump inhibitor lyophilized powder is dissolved in a special solvent. During the configuration process, keep the pH weakly alkaline.
  • the above ingredients are dissolved in distilled water, configured to a mass concentration of 3%, 2.5ml per bottle, and sterilized.
  • Example 21 to Example 25 all formed colloidal films.
  • mice Healthy rats and mice were given a single oral gavage of 30 ml of the preparations prepared according to Examples 1 to 5, and there was no obvious acute toxicity reaction, and no animal death was caused.
  • the gavage dosage is 100 times the clinical dosage.
  • the target organs for acute toxicity are liver and spleen.
  • Test Example 16 The protective effect of Test Example 16 on the gastric ulcer caused by Helicobacter pylori in mice (the preparations obtained in Examples 21-25).
  • the protective effect of Test Example 16 on the gastric ulcer caused by Helicobacter pylori in mice (the preparations obtained in Examples 21-25). 1. Establishment and grouping of the ulcer model. Helicobacter pylori NCTC11637 was inoculated into the broth broth, quickly put into the microaerobic bag, 35°C, cultivated for 72h. Select 75 ICR mice, half male and half, and use random number table to divide 110 mice into experimental group (95) and control group (15). Each mouse in the experimental group was perfused with 200 ⁇ l of Hp bacterial solution (containing 5 ⁇ 108 CFU of bacteria) at an interval of 2 days, and infected 3 times.
  • Hp bacterial solution containing 5 ⁇ 108 CFU of bacteria
  • mice in the experimental group were randomly sacrificed, and gastric antrum tissues were taken for uremycin, bacteriological smear and histological examination to determine whether they were successfully infected with Hp.
  • mice successfully infected with Hp were randomly divided into 6 groups, 15 in each group: model group (gave an equal volume of normal saline), Example 1 group (gavage Example 21, administration dose 0.3ml/ 20g), Example 2 group (gavage to give Example 22, dosage 0.3ml/20g), Example 3 group (gavage to give Example 23, dosage 0.3ml/20g), Example 4 Group (Gavage Example 24, dosage 0.3ml/20g), Example 4 group (Gavage Example 25, dosage 0.3ml/20g). 10 of the control group served as a blank control group (an equal volume of saline was given by gavage). The administration was continued for 7 days, and the animals were sacrificed at the second week after the last administration. The gastric antrum tissue was taken for uremycin, bacteriological smear and histological examination to determine whether Hp was present.
  • Example 1 group, Example 2 group, Example 3 group, Example 4 group, Example 5 group The control group received free diet and water. Except for the control group, the other groups caused 2 wounds on the mucosal surface of the gastric wall through gastroscopy, and performed adequate hemostasis. That is, water control group (equal volume of water gavage), Example 1 group (0.3ml/20g), Example 2 group (0.3ml/20g), Example 3 group (0.3ml/20g), Example 4 group ( 0.3ml/20g), Example 5 group (0.3ml/20g). Gave the stomach once a day for 14 consecutive days, and fast for 48 hours after gavage on the 14th day, can not help but water.
  • mice were sacrificed by pulling the neck uniformly, laparotomy, the pylorus and the pylorus were ligated, and 1.0ml of 1% formaldehyde solution was injected into the stomach.
  • the stomach was fixed in the same concentration of formaldehyde solution for 30 minutes. After the stomach was taken out, it was cut along the greater curvature of the stomach. Pour the contents out of the stomach and rinse gently with water to remove the residue in the stomach. Observe the degree of mucosal ulcer in the glandular stomach, and divide it into healed, effective and ineffective according to the degree of ulcer in mice.
  • mice with hydrochloric acid experimental gastric ulcer (the preparations obtained in Examples 21-25).
  • the mouse is 70, weighing 18-20 grams, half male and half male. Randomly divided into 7 groups, each group of 10, namely control group, model group, implementation
  • the mouse ulcer surface is well improved, and the affected area of the ulcer surface is more than 1/2 intact;
  • Test Example 18 The protective effect of Test Example 18 on experimental gastric ulcer in dogs.
  • each group of the example was sprayed 0.3ml of preparation through gastroscope.
  • the comparative example 1 group was intragastrically administered 0.3ml of the preparation of Example 1 daily
  • the comparative example 2 group was intragastrically administered 0.3ml of the preparation of Example 2 daily
  • the comparative example 3 group was intragastrically administered 0.3ml daily of Example 3. The preparation was administered continuously for 7 days.
  • the mouse ulcer surface is well improved, and the affected area of the ulcer surface is more than 1/2 intact;
  • the preparation made by the invention has obvious therapeutic effect on gastric ulcer.
  • the preparation of the present invention is sprayed through the gastroscope, and the therapeutic effect is significantly improved.
  • the sixth part promotes the proliferation and migration of mucosal epithelial cells in the digestive tract
  • the above ingredients are dissolved in 500ml of 0.01mol of phosphate solution adjusted by 1mol of sodium hydroxide to adjust pH9.0, 5ml per bottle, and high temperature steam sterilization.
  • the solidifying liquid is made of calcium gluconate into a 2% aqueous solution, packaged in 5ml per bottle, and sterilized by high-temperature steam.
  • the above ingredients are dissolved in 600ml of 0.01mol pH9.0 phosphate solution adjusted by 1mol sodium hydroxide, 5ml per bottle, and high temperature steam sterilization.
  • the solidification liquid is prepared with calcium lactate into a 5% aqueous solution, packaged in 5ml per bottle, and sterilized by high-temperature steam.
  • the above ingredients are dissolved in 60ml pure water at 4°C, completely dissolved into a transparent liquid, and then the volume is fixed to 100ml, and each bottle is packed in 5ml. High temperature steam sterilization. Store at low temperature.
  • the solidification liquid is made into 1.1% aqueous solution with calcium chloride, packaged in 5ml per bottle, and sterilized by high temperature steam.
  • the solidification liquid is prepared with calcium lactate into a 5% aqueous solution, packaged in 5ml per bottle, and sterilized by high-temperature steam.
  • the above ingredients are dissolved in distilled water, configured to a mass concentration of 3%, 2.5ml per bottle, and sterilized.
  • Test Example 19 Protective glue on the proliferation and migration of gastric mucosal epithelial cells
  • human gastric mucosal epithelial cells in the logarithmic growth phase are seeded in a 24-well plate with 6 ⁇ 10 5 cells per well, 400 ⁇ L per well, and placed in a 37°C, 5% CO 2 incubator for 24 hours. After tightly connected into a single layer, a capillary with a diameter of 1.5 mm is used to draw a mark vertically in the center of the petri dish, and both sides of the wound can be seen under the microscope.
  • test results show that the examples can obviously promote the proliferation of gastric mucosal cells, and the proliferation speed is more than 1.5 times that of the blank.
  • test results show that the examples can significantly promote the migration of gastric mucosal cells, and the migration speed is more than twice the blank.
  • the research results show that the examples can significantly promote the proliferation and migration of gastric mucosal epithelial cells, and therefore promote the healing of the digestive tract mucosa by accelerating the proliferation and migration of mucosal cells to the wound surface.

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Abstract

一种消化道黏膜保护胶、含有消化道黏膜保护胶的药物组合物和消化道隆起注射剂及消化道黏膜保护胶在制备药物中的应用。消化道黏膜保护胶包括保护层和生物粘合剂。保护层中的有效成分为成胶物质、水溶性钙盐和钙离子络合剂。成胶物质为果胶、海藻酸盐、去乙酰结冷胶中的一种或多种。生物粘合剂为羧甲基纤维素、羧甲基壳聚糖、卡波姆、乙基纤维素、羟丙基纤维素、羟丙基甲基纤维素、白芨多糖、多聚赖氨酸、贻贝粘性蛋白、胶原蛋白、聚多巴胺、透明质酸钠、壳聚糖季铵盐中的一种或多种。消化道黏膜保护胶的pH值不小于7，溶液状态下为中性或偏碱性，产品不形成凝胶状，而在酸性条件下形成凝胶膜。生物粘合剂提供粘性将凝胶膜附着于消化道黏膜上，阻断胃酸和胃蛋白酶对黏膜的消化作用，起到保护黏膜的作用。

Description

消化道黏膜保护胶 技术领域
本发明涉及生物医药技术领域，特别是涉及一种消化道黏膜保护胶。
背景技术
内窥镜下胃黏膜切除手术和胃黏膜剥离术是早期胃癌和胃良性肿瘤切除的微创手术，不仅可达到根治早期胃癌的目的，且具有创伤小、对患者生存质量影响小等优点，已逐渐取代部分传统外科手术。食道和结肠的早期癌变和良性肿瘤切除的微创手术也取代部分的传统外科手术。但是，目前这种手术后创面仅做止血处理，尚无良好的创面保护措施。裸露的创面在消化液的作用下经历溃疡活动期、愈合期和瘢痕期，并且创面的愈合时间也较长。临床研究资料显示病人经内窥镜下胃黏膜切除手术和胃黏膜剥离术6周的愈合率为69％左右。因此，有必要提供一种可以直接通过胃镜应用到胃损伤黏膜表面的产品。
海藻酸盐作为优良的药物制剂基质，医药上多用作亲水性乳化剂、凝胶剂和增稠剂。中国专利[公开号：CN101933894]公开了一种胃黏膜保护胶，包括成胶物质，酸碱调节剂、交联剂和粘合剂；所述成胶物质，交联剂和粘合剂重量比为1∶0.01-0.3∶0.1-0.5，酸碱调节剂的用量为调pH在6.5-8.5。该专利仅利用纯物理学原理保护胃黏膜，使用的交联剂为酸溶解性钙，这种钙在中性溶液中不溶解呈颗粒状态，适合口服使用，不适合通过注射器注射，还有就是这个产品采用的粘附剂为阴离子聚合物和中性聚合物，而不是粘合剂，胃损伤粘膜表面也是带阴离子，因此不能很好的粘合到胃黏膜的创面上，非常容易脱落，不能充分发挥胃黏膜的保护作用。
发明内容
针对上述技术领域中的不足，本发明提供一种全新的采用物理原理并且具有很强粘合力的保护消化道粘膜的方法和产品，起到保护消化道黏膜的作用。
本发明提供一种消化道黏膜保护胶，包括保护层和生物粘合剂；其中保护层的有效成分为成胶物质，水溶性钙盐和钙离子络合剂；成胶物质为果胶、海藻酸盐、去乙酰结冷胶中的一种或多种；生物粘合剂为羧甲基纤维素、羧甲基壳聚糖、卡波姆、乙基纤维素、羟丙基纤维素、羟丙基甲基纤维素、白芨多糖、多聚赖氨酸、贻贝粘性蛋白、胶原蛋白、聚多巴胺、透明质酸钠、壳聚糖季铵盐中的一种或多种；所述消化道黏膜保护胶的pH值不小于7。
进一步，钙离子络合剂为柠檬酸、柠檬酸盐、EDTA或EDTA钠盐中的一种或几种。
进一步，所述果胶为低甲氧基果胶和酰胺化低甲氧基果胶中的一种或两种。
进一步，所述海藻酸盐为海藻酸钠或海藻酸钾中的一种或两种。
进一步，钙盐溶液中钙盐的质量浓度为0.25％-5％。
进一步，成胶物质与钙离子络合剂重量比为3-6:1。
本发明一种实施方式中，所述生物粘合剂为多聚赖氨酸，成胶物质与多聚赖氨酸的重量比为3：1-3。
多聚赖氨酸一般常见的分子量有70，000～150，000，150，000～300，000和>300，000，分子量越大，黏附力越强，固化效果更好，但相对完全溶解较困难。本发明可根据溶解剂等综合选择所需多聚赖氨酸。
本实施方式中，成胶物质，水溶性钙盐、钙离子络合剂重量比为3-6：0.2-1.5：1。
保护胶中还包括溶解剂和酸碱调节剂，溶解剂用于溶解凝胶和生物粘合剂；酸碱调节剂用于调节保护胶的pH值不小于7。
本发明的溶解剂为为水或者其他人体能接受的盐，如磷酸盐溶液。
所述酸碱调节剂为氢氧化钠、氢氧化钾。
本发明另一种实施方式中，所述生物粘合剂为含有贻贝粘蛋白的水溶液。利用含贻贝蛋白粘合层的自粘特性，将胃损伤黏膜和胃黏膜保护胶紧密连接在一起。
该组合中，成胶物质、水溶性钙和钙络合剂的重量比为3-6：0.2-1.5：1，且成胶物质、水溶性钙和钙络合剂的总重量占保护层组分的2.5-3.5wt％。
本发明一个实施例中所述成胶组分为果胶、海藻酸盐、水溶性钙和钙络合剂。果胶、海藻酸盐、水溶性钙和钙络合剂的重量比例为2-3:1-2:0.2-1.5:1。
进一步，贻贝粘蛋白为生物粘合剂的5-25wt％。
进一步，保护层组分中还含有0.3-5wt％的增塑剂。
进一步，增塑剂为甘油、聚乙二醇、聚乙烯醇、甘露醇和山梨醇中的一种或几种。
本发明中，粘合层组分的pH调节剂为乙酸、盐酸、碳酸、柠檬酸、苹果酸、葡萄糖酸、乳酸、草酸、抗坏血酸、酒石酸和苯甲酸中的一种或几种，保护层组分的pH调节剂为碳酸氢钠、碳酸钠、碳酸钾、磷酸氢二钾、磷酸氢二钠、磷酸三钠、磷酸三钾，柠檬酸钠、柠檬酸钾、氢氧化钠、氢氧化钾和柠檬酸一钠中的一种或几种。
本发明还有一种实施方式中，所述消化道黏膜保护胶还包括温度敏感性生物材料，凝胶化 温度调节剂。
进一步，温度敏感性生物材料：凝胶化温度调节剂：生物粘附剂：成胶物质的重量百分比为1-30：0.5-5：0.3-5：0.2-1。
进一步，所述温度敏感性生物材料为聚N-异丙基丙烯酰胺及其衍生物、羟丁基壳聚糖及其衍生物、泊洛沙姆407、聚乳酸/羟基乙酸/聚乙二醇共聚物、聚乙二醇单甲醚-(癸二酸-D,L-乳酸)聚酯酸酐-聚乙二醇单甲醚三嵌段共聚物、纤维素及其衍生物和聚乙二醇/聚己内酯嵌段共聚物中的一种或多种。
进一步，所述的凝胶化温度调节剂为泊洛沙姆188、聚山梨醇酯、聚乙二醇中的一种或多种。
胶体液的成分溶解于溶解液中，形成胶体溶液；保护胶的相转变温度25℃-40℃。
溶解液为纯水、生理盐水和磷酸盐溶液中的一种或多种。
本发明中所述保护胶还包括固定液，固定液为水溶性钙溶液。
进一步，所述水溶性钙为氯化钙、乳酸钙、葡萄糖酸钙中的一种或多种。
进一步，水溶性钙溶液浓度0.3％-5％。
本发明提供的其他实施方式中，所述保护胶还包括治疗溃疡、肿瘤的有效药物量的药物化合物或组合物。
本发明还提供一种保护胶还包括胃酸抑制剂、抗幽门杆菌制剂。
其中，胃酸抑制剂为质子泵抑制剂或H 2受体拮抗剂。所述的胃酸抑制剂为质子泵抑制剂时,需添加pH调节剂。
进一步，抗幽门杆菌制剂由铋剂、甲硝唑或替硝唑、阿莫西林、克拉霉素、四环素、呋喃唑酮中的一种或多种组成。
进一步，胃酸抑制剂、抗幽门杆菌制剂和保护胶其他组分的重量比为0.001-0.05:0.001-1:1-20。
进一步，消化道黏膜保护胶还包括促进消化道黏膜上皮细胞增殖和迁移的成分，包括甘草多糖、β-葡聚糖、党参多糖、索拉胶、杏鲍菇葡聚糖、燕麦葡聚糖、灰树花多糖、谷物葡聚糖、酵母葡聚糖、灵芝多糖中的一种或多种。这些成分在整个保护胶中的重量比0.001％-0.5％。
本发明还提供一种药物组合物，含有前述的消化道黏膜保护胶和其他具有治疗效果的药物。本发明所指具有治疗效果的药物可以为各种治疗消化道疾病的药物。
本发明还提供一种消化道隆起注射剂，包含前述的消化道黏膜保护胶。
将所述消化道粘膜下注射隆起剂注射至粘膜下方，使粘膜层和肌肉层分离，方便粘膜剥离。
本发明还提供前述的消化道黏膜保护胶在植被治疗消化性溃疡、应激性溃疡、溃疡性结肠炎、胃炎、胃黏膜切除和剥离术后的创面、食道粘膜剥离术和肠道粘膜剥离术后的创面、胃泌素瘤和胃癌药物中的应用。
由于消化道基本处于酸性或弱酸性环境，因此本发明的目的均能实现。本发明在描述效果时，主要以胃部环境进行描写，但是并不代表本发明仅局限于胃部，本发明可以应用于人体所有消化道，包括食道，胃，肠等。
与现有技术相比，本发明具有以下优点：
本发明采用科学原理，在胃酸条件下，使成胶物质在胃肠粘膜表面形成胶体膜，阻断胃酸和胃蛋白酶对粘膜的消化作用,促进溃疡愈合。
本发明的消化道黏膜保护胶为自固化自粘合消化道损伤粘膜保护胶产品在使用前，溶液状态下为中性或偏碱性质，产品不形成凝胶状态。而在酸性条件下，使本品成凝胶膜，而生物粘合剂提供粘结于胃粘膜的性能，使凝胶膜附着于胃粘膜上，阻断胃酸和胃蛋白酶对粘膜的消化作用，起到保护胃粘膜的作用。
本发明基于成交物质果胶和海藻酸盐的离子凝固机理，在偏碱性环境条件下，钙离子被钙离子络合剂络合，因此不能与成胶物质交联。和胃酸接触作用后，钙离子在酸性环境和柠檬酸离子不能处于络合状态，钙离子释放出来，使果胶或海藻酸盐发生交联而形成凝胶膜。
保护胶是采用从植物提取的天然高分子材料海藻酸钠和/或果胶以及交联剂制成，通过胃窥镜喷洒在胃损伤粘膜表面形成胶体膜，起到保护胃黏膜的作用。本产品的特点是在通常情况下为液体，在胃酸的作用下，保护胶中的交联剂释放出来，在胃损伤粘膜表面形成胶体膜，避免胃酸和胃蛋白酶对胃肠粘膜的消化作用，促进溃疡的愈合。
同时采用的粘合剂如多聚赖氨酸等，在偏碱性的溶液中不和成胶物质作用，为液体状态。在胃酸的环境下表现为阳离子聚合物的特性，一是和成胶物质交联，增强的保护胶的强度，二是还可以通过其阳离子的特性和胃粘膜损伤表面的阴离子结合，粘合到创伤表面。
由于粘膜保护胶涂到损伤粘膜后不会马上和胃酸接触，会有流动的问题，所以本发明采用固化液使得保护胶迅速固定在损伤粘膜表面，避免了保护胶的流动问题。
进一步的，本发明提供了自固化双组分离子和温度双敏感型消化道损伤粘膜保护胶具有离子和温度双敏感。
自固定双组分温度敏感性消化道粘膜保护胶包括胶体液和固定液，胶体液包括温度敏感性 材料，凝胶化温度调节剂、生物粘附剂和离子敏感固定剂，固定液为水溶性钙溶液。
一方面，胶体液在低于室温(25℃)条件下为液体，通过内镜喷涂到消化道黏膜创面后，喷涂固定液迅速固定，而在体温(37℃)条件下，温度敏感性材料在体温下发生相变，使本品由液体通过相变变成凝胶，而粘附剂提供粘结于损伤粘膜的性能，使凝胶附着于粘膜上不易脱落，阻断消化液对粘膜的消化作用，起到保护损伤粘膜的作用。
另外，固化液中的钙离子与胶体液中的离子敏感固定剂形成凝胶膜，避免胶体液的流失，也加强对损伤粘膜的保护。
具体实施方式
下面结合实施例对本发明作进一步的详细说明。
本发明实施例中没有特殊说明，采用的均为一般化学品，市场上购买即可得到。
第一部分，粘合剂为多聚赖氨酸。
其中实施例中综合考虑溶解难度选择的多聚赖氨酸的分子量150,000～300,000的多聚赖氨酸。其他分子量的多聚赖氨酸也可以，只要实现溶解即可。
实施例1
1.海藻酸钾12克
2.氯化钙1克
3.柠檬酸钠2克
4.多聚赖氨酸10g
__________________________________________________________________________
以上成份溶解在0.01mol采用1mol氢氧化钠调pH9.0磷酸盐溶液500ml中溶解，每瓶5ml包装,高温蒸汽灭菌。固化液采用氯化钙配制成1.1％的水溶液，每瓶5ml包装,高温蒸汽灭菌。
实施例2
1.酰胺化低甲氧基果胶15克
2.氯化钙1克
3.柠檬酸钠2.5克
4.多聚赖氨酸12g
_____________________________________________________________________________
以上成份溶解在0.01mol采用1mol氢氧化钠调pH9.0磷酸盐溶液600ml中溶解，每瓶5ml包 装,高温蒸汽灭菌。固化液采用氯化钙配制成1.1％的水溶液，每瓶5ml包装,高温蒸汽灭菌。
实施例3
1.海藻酸钠11克
2.柠檬酸钠4克
3.氯化钙1.5克
4.多聚赖氨酸10g
__________________________________________________________________________
以上成份溶解在0.01mol采用1mol氢氧化钠调pH9.0磷酸盐溶液500ml中溶解，每瓶5ml包装,高温蒸汽灭菌。固化液采用葡萄糖酸钙配制成2％的水溶液，每瓶5ml包装,高温蒸汽灭菌。
实施例4
1.海藻酸钠11克
2.柠檬酸钠4克
3.葡萄糖酸钙6克
4.多聚赖氨酸10g
___________________________________________________________________
以上成份溶解在0.01mol采用1mol氢氧化钠调pH9.0磷酸盐溶液600ml中溶解，每瓶5ml包装,高温蒸汽灭菌。固化液采用氯化钙配制成1.7％的水溶液，每瓶5ml包装,高温蒸汽灭菌。
实施例5
1.海藻酸钾10克
2.柠檬酸钠4克
3.乳酸钙2.8克
4多聚赖氨酸10g
_________________________________________________________________
以上成份溶解在0.01mol采用1mol氢氧化钠调pH9.0磷酸盐溶液600ml中溶解，每瓶5ml包装,高温蒸汽灭菌。固化液采用乳酸钙配制成5％的水溶液，每瓶5ml包装,高温蒸汽灭菌。
实验例效果评价
以上实施例的液体的直接用于试验。
1体外胶体膜形成试验
分别取上述液体2ml涂布在新鲜的猪皮内侧面上，然后再喷洒固化液，观察胶体膜形成的情况。结果是实施例1到实施例5均形成胶体膜。
测试例2
安全性研究试验
将实施例1-5进行以下生物学试验
1)口腔粘膜刺激：将12ml胶体液倒入直径15cm的无菌培养皿中，然后倒入固定液后30分钟，用生理盐水冲吸收掉多余的固定液，然后按照3cm 2/ml比例加入生理盐水，在37摄氏度浸提72小时制备试验液。将样品的实验液制成直径不大于5mm棉球浸透置于3只金黄地鼠一侧颊囊中。接触时间每次最少5min，每天一次，共4次，末次接触后24h肉眼观察颊囊,无痛处死地鼠,取颊囊有代表性部位的组织样品放入4％甲醛溶液中固定后并制作组织切片后进行组织学评价。结果刺激指数均为0，被试样品无口腔粘膜刺激性。
2)细胞毒性：将12ml胶体液倒入直径15cm的无菌培养皿中，然后倒入固定液后30分钟，用生理盐水冲吸收掉多余的固定液，然后按照3cm 2/ml比例加入培养基，在37摄氏度浸提24小时制备试验液。然后按GB/T16886.5中规定的细胞毒性试验采用MTT法测定，细胞毒性均在0-1级范围内。
3)致敏试验：将12ml胶体液倒入直径15cm的无菌培养皿中，然后倒入固定液后30分钟，用生理盐水冲吸收掉多余的固定液，然后按照3cm 2/ml比例加入生理盐水，在37摄氏度浸提72小时制备试验液。然后按GB/T16886.10中规定的方法进行皮肤致敏试验，均未观察到致敏作用。
测试例3
组织粘附力的测定(组织留存量法)：
将实施例1-5分别进行以下试验
取SD大鼠,禁食24h用戊巴比妥钠溶液(40mg/kg)经腹腔注射麻醉,解剖取出胃置生理盐水(37℃)中切取胃,用生理盐水将胃内壁清洗干净，清洗的胃在2小时内使用。剪取一定面积的胃组织(2cm×2cm),固定于聚乙烯薄膜上，将0.5ml保护胶均匀涂布于胃黏膜创面上，然后喷涂固化液。胃组织在相对湿度为92.5％的恒湿密闭容器中放置20分钟，将经上述处理的胃组织固定于冲洗斜槽上,将斜槽的角度调至60度，调节蠕动泵流速为20ml/min,将胃组织用 0.1mol/L盐酸冲洗5mins,冲洗液收集于一已知重量的烧杯中,在70℃烘干,称重,组织粘附力用粘附百分率表示。
计算方法如下:
胃组织粘附百分率(Bg/％)Bg/％＝{[M-(G-g-m)]/M}x100％
式中M为黏膜保护胶的重量(0.5ml同条件下烘干)；g为空烧杯重；G为烧杯与烘干后残渣总重；m为同体积冲洗液所含固体物质的量(空白对照)。B值越大表示粘附力越大。
结果显示：实施例1：100％，实施2：99％，实施例3：100％，实施例4：100％，实施例5：99％。试验表明实施例1-5的消化道黏膜保护胶对组织的粘附力较强，不易脱落。
测试例4
动物试验
动物分两组，体重3kg左右。实验组6只，对照组6只。
手术前24小时禁食，不禁水。
麻醉方法：推荐家兔用质量浓度30g/L戊巴比妥钠静脉注射1.0ml/kg。
家兔仰面固定手术台上，腹部去毛。按外科常规手术要求以2％碘酊和75％乙醇溶液消毒试验区域。
在上腹部逐层切开皮肤、肌肉层和腹膜，如有出血结扎止血。暴露胃部，在胃大弯处切开胃，用生理盐水清洗胃部，在胃体内的胃大弯侧面的黏膜下，1：10000肾上腺素生理盐水进行黏膜下注射，注射1ml生理盐水形成胃黏膜突出，然后用环套器切除直径1cm的黏膜形成创面，喷洒凝血酶(1ml内含凝血酶50U)并压迫止血完全，测量创面的直径。对照组不做任何处理，实验组涂布0.5ml保护胶，再喷涂固化液后形成固体化的保护膜，然后缝合胃部。再逐层缝合腹壁。放饲养笼中，禁食一天。
结果观察：与手术后1周安乐死动物，沿原来的切口切开胃壁，观察创面愈合情况，测量创面的直径，结果发现实验组手术后1周溃疡的实验组6只家兔愈合，愈合率100％；对照组2只愈合，4只未愈合，愈合率33％。以上结果可以看出,按实施例1能明显促进的胃黏膜损伤性溃疡愈合，对胃溃疡有明显的治疗作用。
第二部分，粘合剂为多聚赖氨酸。
实施例6
粘合层组分：
贻贝粘蛋白  25g，
氯化钠      0.9g，
以上成份溶解在蒸馏水100ml中，用乙酸调pH＝5.0，每瓶2.0ml包装，辐照灭菌。
保护层组分：
海藻酸钠    7.5g，
氯化钙      0.5g，
柠檬酸钠    2.5g，
以上成分溶解在蒸馏水中，氢氧化钠调酸碱度到8.5，配置成质量浓度为3.0％的溶液，再按每100ml溶液加入甘油1g，每瓶5ml包装,高温蒸汽灭菌。
实施例7
粘合层组分
贻贝粘蛋白  5g，
氯化钠      0.9g，
以上成份溶解在蒸馏水100ml中，用柠檬酸调pH＝5.0每瓶2.0ml包装，辐照灭菌。
保护层组分
果胶      4g，
海藻酸钠  8g，
氯化钙    3g，
EDTA      2g，
以上成份溶解在蒸馏水中，氢氧化钠调酸碱度到8.5，配置成质量浓度为2.5％，并按每100ml加入聚乙烯醇5g，每瓶5ml包装,高温蒸汽灭菌保存。
实施例8
粘合层组分
贻贝粘蛋白  10g，
氯化钠      0.9g，
以上成份溶解在蒸馏水100ml中，用乙酸调pH＝5.0,每瓶2.0ml包装，辐照灭菌。
保护层组分
海藻酸钠      10g，
葡萄糖酸钙    3g，
柠檬酸钠：    2.5g，
氢氧化钠(pH调节剂)：调节pH为8.0，
配置过程中，保持pH为弱碱性。以上成份溶解在蒸馏水中，配置成质量浓度为3.5％，并按每100ml加入甘露醇0.15g和山梨醇0.15g，每瓶5ml包装，高温蒸汽灭菌保存。
实施例9
粘合层组分
贻贝粘蛋白  25g，
氯化钠      0.9g，
以上成份溶解在蒸馏水100ml中，用柠檬酸调pH＝5.0,每瓶2.0ml包装，辐照灭菌。
保护层组分
海藻酸钾    10g，
乳酸钙      1.4g，
氯化钙      0.5g，
柠檬酸钠：  2.5g，
氢氧化钠(pH调节剂)适量：调节pH为9.0，
配置过程中，保持pH为弱碱性。以上成份溶解在蒸馏水中，配置成质量浓度为3％，并按每100ml加入聚乙二醇1g，每瓶5ml包装,高温蒸汽灭菌保存。
实施例10
粘合层组分
贻贝粘蛋白  12g，
氯化钠      0.9g，
以上成份溶解在蒸馏水100ml中，用乙酸调pH＝5.0,每瓶2.0ml包装，辐照灭菌。
保护层组分
海藻酸钠    10g，
乳酸钙      1.4g，
柠檬酸钠：  2.5g，
氢氧化钠(pH调节剂)：调节pH为8.5
配置过程中，保持pH为弱碱性。以上成份溶解在蒸馏水中，配置成质量浓度为3％，并按每100ml加入甘油1g，每瓶5ml包装,高温蒸汽灭菌保存。
应用例1
用胃窥镜将粘合层组分喷涂到胃黏膜创面上，之后再用胃窥镜将保护层组分喷涂到已经喷涂了粘合层组分的胃黏膜创面上。
胃黏膜创面为消化性溃疡、应激性溃疡、胃炎、胃黏膜切除和剥离术后的创面。
需要说明的是，创面深度以0.5cm为基准，粘合层组分按照创面面积的0.2-0.25倍的量进行喷涂，即1cm2的创面喷涂0.2-0.25mL的粘合层组分。保护层组分按照创面面积的0.3-0.5倍的量进行喷涂，即1cm 2的创面喷涂0.3-0.5mL的粘合层组分。
当创面深度小于或大于基准值时，粘合层组分和保护层组分的用量按创面实际深度与基准深度的比例进行相应调整。
本实施例自粘性胃黏膜保护胶粘合层组分喷涂到创面后形成粘合层，保护层组分喷涂到粘合层后形成保护层。
第一界面：保护层和粘合层之间的界面，通过贻贝粘蛋白的阳离子特性和海藻酸的阴离子结合。
第二界面：粘合层和胃黏膜创面的交界面，通过贻贝粘蛋白的阳离子特性与创面组织细胞的阴离子结合和在生理条件下贻贝粘蛋白的部分疏水基团暴露出来与细胞膜的脂质双分子层通过疏水作用相互结合。
粘合层：粘合层组分中贻贝黏蛋白多巴基团中的一些酚羟基氧化成醌，氧化的多巴和未氧化的 多巴交联形成高分子网状 聚合物。
保护层：在胃酸的作用下通过离子交联固化形成保护膜。
具体的说，粘合层组分首先涂覆到损伤黏膜表面，也即创面上，形成粘合层，由于粘合层中贻贝粘蛋白具有带正电荷的特性和创面带负电荷的细胞组织牢固结合，另外粘合层贻贝粘蛋白的几种氨基酸含有 疏水基团，在生理溶液存在情况下，贻贝粘蛋白的部分疏水基团暴露出来，也可与细胞膜的脂质双分子层通过疏水作用相互结合。
粘合层的正电荷也能和保护层中带负电荷的海藻酸盐通过离子键结合在一起。由于保护层为碱性，通过扩散到粘合层使pH升高的情况下，粘合层中贻贝黏蛋白多巴基团中的一些酚羟基氧化成醌，这一过程在体内儿茶酚 氧化酶存在下会被加速，氧化的多巴和未氧化的 多巴交联，粘合层形成高分子网状 聚合物。
粘合层的酸性也会扩散到保护层，使保护层也发生内部交联，这样就可以紧密的把损伤 的胃黏膜和保护层连接在一起，使得保护层比较牢固的保持在创面表面，起到保护胃损伤黏膜表面的作用。保护层在胃酸的作用下通过离子交联固化形成保护膜，另外添加的增塑剂使得保护膜有一定的弹性，具有耐胃蠕动的性能。
测试例5
体外胶体膜形成测试
分别取上述粘合层组分1ml和保护层组分液体2ml涂布在新鲜的猪胃的内侧面上，然后再喷洒含有稀盐酸的人工胃液，观察胶体膜形成的情况。结果是实施例6-10均形成胶体膜。
测试例6
安全性研究试验
将实施例6-10进行以下生物学试验
1)皮内刺激：将样品的实验液注射0.2ml于皮内，注射后2小时、24小时和72小时观察水肿及红肿情况，所有实施例的反应在0-1级。
2)细胞毒性：将样品按GB/T16886.5中规定的细胞毒性试验采用MTT法测定，细胞毒性均在0-1级范围内。
3)致敏试验：将样品按GB/T16886.10中规定的方法进行皮肤致敏试验，均未观察到致敏作用。
测试例7
组织粘附力的测定(组织留存量法)：
将实施例6-10分别进行以下试验
取SD大鼠,禁食24h用戊巴比妥钠溶液(40mg/kg)经腹腔注射麻醉,解剖取出胃置生理盐水(37℃)中切取胃,用生理盐水将胃内壁清洗干净，清洗的胃在2小时内使用。剪取一定面积的胃组织(2cm×2cm),固定于聚乙烯薄膜上，将0.3ml粘合层组分涂布到创面上，然后在将0.5ml保护层组分均匀涂布于胃黏膜创面上。胃组织在相对湿度为92.5％的恒湿密闭容器中放置20分钟，将经上述处理的胃组织固定于冲洗斜槽上,将斜槽的角度调至60度，调节蠕动泵流速为20ml/min,将胃组织用0.1mol/L盐酸冲洗5mins,冲洗液收集于一已知重量的烧杯中,在70℃烘干,称重,组织粘附力用粘附百分率表示。
计算方法如下:
胃组织粘附百分率(Bg/％)Bg/％＝{[M-(G-g-m)]/M}x100％
式中M为黏膜保护胶的重量(0.5ml同条件下烘干)；g为空烧杯重；G为烧杯与烘干后残渣总重；m为同体积冲洗液所含固体物质的量(空白对照)。B值越大表示粘附力越大。
结果显示：实施例1：98％，实施2：99％，实施例3：97％，实施例4：95％，实施例5：99％。试验表明实施例6-10的消化道黏膜保护胶对组织的粘附力较强，不易脱落。
测试例9
动物试验
动物分两组，体重3kg左右。实验组6只，对照组6只。
手术前24小时禁食，不禁水。
麻醉方法：推荐家兔用质量浓度30g/L戊巴比妥钠静脉注射1.0ml/kg。
家兔仰面固定手术台上，腹部去毛。按外科常规手术要求以2％碘酊和75％乙醇溶液消毒试验区域。
在上腹部逐层切开皮肤、肌肉层和腹膜，如有出血结扎止血。暴露胃部，在胃大弯处切开胃，用生理盐水清洗胃部，在胃体内的胃大弯侧面的黏膜下，1：10000肾上腺素生理盐水进行黏膜下注射，注射1ml生理盐水形成胃黏膜突出，然后用环套器切除直径1cm的黏膜形成创面，喷洒凝血酶(1ml内含凝血酶50U)并压迫止血完全，测量创面的直径。对照组不做任何处理，实验组先涂布0.2ml粘合层组分，然后再涂布0.4ml保护层组分，可见保护层组分接触胃酸后形成固体化的保护膜，然后缝合胃部。再逐层缝合腹壁。放饲养笼中，禁食一天。
结果观察：与手术后1周安乐死动物，沿原来的切口切开胃壁，观察创面愈合情况，测量创面的直径，结果发现实验组手术后1周溃疡的实验组5只家兔愈合，1只未愈合，愈合率83％；对照组2只愈合，4只未愈合，愈合率33％。以上结果可以看出,按实施例1能明显促进的胃黏膜损伤性溃疡愈合，对胃溃疡有明显的治疗作用。
第三部分，粘合剂为多聚赖氨酸，同时添加温度敏感材料。
实施例11
1.泊洛沙姆407 18克
2.泊洛沙姆188 0.5克
3.羟丙基甲基纤维素 0.5克
4.海藻酸钠 0.3克
以上成份在4℃溶解在60ml纯水中，溶解完全呈透明液体，然后定容到100ml，每瓶5ml包装。高温蒸汽灭菌。低温保存。
固化液采用氯化钙配制成1.1％的水溶液，每瓶5ml包装,高温蒸汽灭菌。
实施例12
1.泊洛沙姆407 20克
2.泊洛沙姆188 1克
3.羟丙基甲基纤维素 0.5克
4.海藻酸钠 0.4克
以上成份在4℃溶解在60ml纯水中，溶解完全呈透明液体，然后定容到100ml，每瓶5ml包装,高温蒸汽灭菌。低温保存。
固化液采用氯化钙配制成1.1％的水溶液，每瓶5ml包装,高温蒸汽灭菌。
实施例13
1.羟丁基壳聚糖 5克
2.泊洛沙姆188 0.5克
3.胶原蛋白 2克
4.酰胺化低甲氧基果胶 1克
以上成份在4℃溶解在60ml纯水中，溶解完全呈透明液体，然后定容到100ml，每瓶5ml包装,高温蒸汽灭菌。低温保存。
固化液采用葡萄糖酸钙配制成2％的水溶液，每瓶5ml包装,高温蒸汽灭菌。
实施例14
1.羟丁基壳聚糖 8克
2.泊洛沙姆188 0.5克
3.胶原蛋白 2克
4.去乙酰结冷胶 0.2克
以上成份在4℃溶解在60ml纯水中，溶解完全呈透明液体，然后定容到100ml，每瓶5ml 包装,高温蒸汽灭菌。低温保存。
固化液采用氯化钙配制成1.7％的水溶液，每瓶5ml包装,高温蒸汽灭菌。
实施例15
1.聚N-异丙基丙烯酰胺 10克
2.泊洛沙姆188 0.5克
3.胶原带白 2克
4.去乙酰结冷胶 0.2克
以上成份在4℃溶解在60ml生理盐水中，溶解完全呈透明液体，然后定容到100ml，每瓶5ml包装,高温蒸汽灭菌。低温保存。
固化液采用乳酸钙配制成5％的水溶液，每瓶5ml包装,高温蒸汽灭菌。
实验例效果评价
以上液体直接用于试验。
测试例10
体外胶体膜形成试验
分别取上述液体2ml涂布在预温到37℃新鲜的猪皮内侧面上，喷涂固定液，然后放置在37℃恒温箱中10分钟，观察胶体膜形成的情况。结果是实施例11到实施例15均形成胶体膜。
测试例11
安全性研究试验
将实施例11-15进行以下生物学试验
1)口腔粘膜刺激：将5ml胶体液倒入直径9cm的无菌培养皿中，喷涂固定液，然后放置在37℃恒温箱中30分钟使其成膜，然后按照3cm 2/ml比例加入生理盐水，在37℃浸提72小时制备试验液。将样品的实验液制成直径不大于5mm棉球浸透置于3只金黄地鼠一侧颊囊中。接触时间每次最少5min，每天一次，共4次，末次接触后24h肉眼观察颊囊,无痛处死地鼠,取颊囊有代表性部位的组织样品放入4％甲醛溶液中固定后并制作组织切片后进行组织学评价。结果刺激指数均为0，被试样品无口腔粘膜刺激性。
2)细胞毒性：将5ml胶体液倒入直径9cm的无菌培养皿中，喷涂固定液，然后放置在 37℃恒温箱中30分钟使其成膜，然后按照3cm 2/ml比例加入培养基，在37℃浸提24小时制备试验液。然后按GB/T16886.5中规定的细胞毒性试验采用MTT法测定，细胞增值率均在76％-89％级范围内。
3)致敏试验：将5ml胶体液倒入直径9cm的无菌培养皿中，喷涂固定液，然后放置在37℃恒温箱中30分钟使其成膜，，然后按照3cm 2/ml比例加入生理盐水，在37℃浸提72小时制备试验液。然后按GB/T16886.10中规定的方法进行皮肤致敏试验，均未观察到致敏作用。
测试例12
组织粘附力的测定(组织留存量法)：
将实施例11-15分别进行以下试验
取SD大鼠,禁食24h用戊巴比妥钠溶液(40mg/kg)经腹腔注射麻醉,解剖取出胃置生理盐水(37℃)中切取胃,用生理盐水将胃内壁清洗干净，清洗的胃在2小时内使用。剪取一定面积的胃组织(2cm×2cm),固定于聚乙烯薄膜上，将0.5ml保护胶均匀涂布于预温到37℃的胃黏膜创面上，喷涂固定液。胃组织在相对湿度为92.5％的恒湿密闭容器中放置20分钟，将经上述处理的胃组织固定于冲洗斜槽上,将斜槽的角度调至60度，调节蠕动泵流速为20ml/min,将胃组织用0.1mol/L盐酸冲洗5mins,冲洗液收集于一已知重量的烧杯中,在70℃烘干,称重,组织粘附力用粘附百分率表示。
计算方法如下:
胃组织粘附百分率(Bg/％)Bg/％＝{[M-(G-g-m)]/M}x100％
式中M为黏膜保护胶的重量(0.5ml同条件下烘干)；g为空烧杯重；G为烧杯与烘干后残渣总重；m为同体积冲洗液所含固体物质的量(空白对照)。B值越大表示粘附力越大。
结果显示：实施例11：90％，实施例12：91％，实施例13：91％，实施例14：92％，实施例15：90％。试验表明实施例11-15的温度敏感型黏膜保护胶对组织的粘附力较强，不易脱落。
测试例13
动物试验
动物分两组，体重3kg左右。实验组6只，对照组6只。
手术前24小时禁食，不禁水。
麻醉方法：推荐家兔用质量浓度30g/L戊巴比妥钠静脉注射1.0ml/kg。
家兔仰面固定手术台上，腹部去毛。按外科常规手术要求以2％碘酊和75％乙醇溶液消毒试验区域。
在上腹部逐层切开皮肤、肌肉层和腹膜，如有出血结扎止血。暴露胃部，在胃大弯处切开胃，用生理盐水清洗胃部，在胃体内的胃大弯侧面的黏膜下，1：10000肾上腺素生理盐水进行黏膜下注射，注射1ml生理盐水形成胃黏膜突出，然后用环套器切除直径1cm的黏膜形成创面，喷洒凝血酶(1ml内含凝血酶50U)并压迫止血完全，测量创面的直径。对照组不做任何处理，实验组涂布0.5ml保护胶，然后喷涂固定液，然后缝合胃部。再逐层缝合腹壁。放饲养笼中，禁食一天。
结果观察：与手术后1周安乐死动物，沿原来的切口切开胃壁，观察创面愈合情况，测量创面的直径，结果发现实验组手术后1周溃疡的实验组5只家兔愈合，1只未愈合，愈合率83％；对照组2只愈合，4只未愈合，愈合率33％。以上结果可以看出,按实施例1能明显促进的胃黏膜损伤性溃疡愈合，对胃溃疡有明显的治疗作用。
第四部分保护胶作为注射隆起剂
实施例16
1.海藻酸钾 12克
2.氯化钙 1克
3.柠檬酸钠 2克
4.多聚赖氨酸 10g
以上成份溶解在采用1mol氢氧化钠调pH8.0生理盐溶液1500ml中溶解，每瓶15ml包装，高温蒸汽灭菌，得到注射隆起剂。
实施例17
1.酰胺化低甲氧基果胶 15克
2.氯化钙 1克
3.柠檬酸钠 2.5克
4.多聚赖氨酸 12g
以上成份溶解在采用1mol氢氧化钠调pH8.0生理盐溶液1500ml中溶解，每瓶5ml包装,高温蒸汽灭菌。
实施例18
1.海藻酸钠 11克
2.柠檬酸钠 4克
3.氯化钙 1.5克
4.多聚赖氨酸 10g
以上成份溶解在采用1mol氢氧化钠调pH8.0生理盐溶液1500ml中溶解，每瓶15ml包装,高温蒸汽灭菌。
实施例19
1.海藻酸钠 11克
2.柠檬酸钠 4克
3.葡萄糖酸钙 6克
4.多聚赖氨酸 10g
以上成份溶解在采用1mol氢氧化钠调pH8.0生理盐溶液1500ml中溶解，每瓶15ml包装,高温蒸汽灭菌。
实施例20
1.海藻酸钾 10克
2.柠檬酸钠 4克
3.乳酸钙 2.8克
4多聚赖氨酸 10g
以上成份溶解在采用1mol氢氧化钠调pH8.0磷酸盐溶液1500ml中溶解，每瓶15ml包装,高温蒸汽灭菌。
实验例效果评价
以上实施例的产品的直接用于试验。
1安全性研究试验
将实施例16-20制备的注射隆起剂进行以下生物学试验
1)口腔粘膜刺激：按照0.2g/ml比例加入生理盐水制备试验液。将样品的实验液制成直径不 大于5mm棉球浸透置于3只金黄地鼠一侧颊囊中。接触时间每次最少5min，每天一次，共4次，末次接触后24h肉眼观察颊囊,无痛处死地鼠,取颊囊有代表性部位的组织样品放入4％甲醛溶液中固定后并制作组织切片后进行组织学评价。结果刺激指数均为0，被试样品无口腔粘膜刺激性。
2)细胞毒性：按照0.2g/ml比例加入培养基制备试验液。然后按GB/T16886.5中规定的细胞毒性试验采用MTT法测定，细胞毒性均在0-1级范围内。
3)致敏试验：按照0.2g/ml比例加入生理盐水制备试验液。然后按GB/T16886.10中规定的方法进行皮肤致敏试验，均未观察到致敏作用。
2黏膜隆起效果试验
1)试验材料：
(1)新鲜离体猪胃3个，购自超市，待用时于4℃冰箱保存，实验时间距离猪屠宰时间不超过12h。
(2)实验台、注射器，计时器，大头针，丝线，照相机，直尺，白色泡沫板。23G、25G(Gauge，简称G)注射针。
2)试验方法
用大头针将猪胃固定在方形泡沫板上，黏膜下注射16-20号实施例的注射隆起剂及生理盐水(对照)溶液6组液体各1ml，猪胃黏膜下立即形成隆起，用直尺联合丝线测量各组隆起的高度(0min)，并用数码相机垂直于胃所在平面拍照，准确记录各组黏膜隆起的高度；5、15、30和60min后分别再在同一位置测量隆起高度，同时用数码相机拍照进行记录。独立重复猪胃黏膜下注射实验各3次。结果表明本发明具有良好的粘膜隆起效果。试验结果见表1.
表1：试验结果
Figure PCTCN2020078733-appb-000001
Figure PCTCN2020078733-appb-000002
第五部分药物组合物
实施例21
药物组合物成分：
Figure PCTCN2020078733-appb-000003
以上成份溶解在蒸馏水中,配置成质量浓度为3％,每瓶2.5ml包装,高温蒸汽灭菌。
实施例22
药物组合物原料:
Figure PCTCN2020078733-appb-000004
以上成份溶解在蒸馏水中,配置成质量浓度为3％,每瓶2.5ml包装,灭菌保存。
实施例23
药物组合物原料:
Figure PCTCN2020078733-appb-000005
小苏打(pH调节剂):调节pH为8.0。
质子泵抑制剂冻干粉由专用溶剂溶解。配置过程中,保持pH为弱碱性。以上成份溶解在蒸馏水中,配置成质量浓度为3％,每瓶2.5ml包装,灭菌保存。
实施例24
药物组合物原料如下：
Figure PCTCN2020078733-appb-000006
小苏打(pH调节剂):调节pH为8.0。
质子泵抑制剂冻干粉由专用溶剂溶解。配置过程中,保持pH为弱碱性。以上成份溶解在蒸馏水中,配置成质量浓度为3％,每瓶2.5ml包装,灭菌保存。
实施例25
药物组合物原料如下：
Figure PCTCN2020078733-appb-000007
小苏打(pH调节剂):调节pH为8.0。
质子泵抑制剂冻干粉由专用溶剂溶解。配置过程中,保持pH为弱碱性。以上成份溶解在蒸馏水中,配置成质量浓度为3％,每瓶2.5ml包装,灭菌保存。
测试例14
体外胶体膜形成测试分别取上述液体2ml涂布在新鲜的猪皮内侧面上,然后再喷洒含有稀盐酸的人工胃液,观察胶体膜形成的情况。结果是实施例21到实施例25均形成胶体膜。
_______________________________________________________________
测试例15安全性研究试验
健康大鼠和小鼠分别单次经口灌胃给予根据实施例1～5制备的制剂30ml,未出现明显的急性毒性反应,也未引起动物死亡。灌胃剂量为临床拟用量的100倍。小鼠未出现明显的急性毒性反应,大鼠出现拉稀便,活动减少等反应,急性毒性靶器官为肝脏和脾脏。
测试例16对小鼠幽门杆菌致胃溃疡的保护作用(实施例21～25所得制剂)。1.溃疡模型的建立及分组把幽门螺杆菌NCTC11637接种到布氏肉汤培养基上,迅速放入微需氧袋,35℃,培养72h。选取75只ICR小鼠,雌雄各半,应用随机数字表将110只小鼠分成实验组(95只)和对照组(15只)。实验组每只小鼠灌喂Hp菌液200μl(含菌5×108CFU)间隔2d,共感染3次。所有动物灌喂处理前禁食24h,禁饮水4h,灌喂后继续禁食、水4h。对照组灌喂等量布氏肉汤培养基。6周后随机处死实验组5只小鼠,取胃窦部组织进行尿霉素、细菌学涂片检查和组织学检查以确定是否成功感染Hp。
将成功感染Hp的90只小鼠随机分成6组,每组15只:模型组(灌胃给予等体积生理盐水)、实施例1组(灌胃给与实施例21,给药剂量0.3ml/20g)、实施例2组(灌胃给与实施例22,给药剂量0.3ml/20g)、实施例3组(灌胃给与实施例23,给药剂量0.3ml/20g)、实施例4组(灌胃给与实施例24,给药剂量0.3ml/20g)、实施例4组(灌胃给与实施例25,给药剂量0.3ml/20g)。对照组10只作为空白对照组(灌胃给予等体积生理盐水)。连续给药7d,于末次给药后第2周处死动物,取胃窦部组织进行尿霉素、细菌学涂片检查和组织学检查以确定是否Hp是否存在。
表2对小鼠幽门杆菌致胃溃疡的实验结果
Figure PCTCN2020078733-appb-000008
各组治疗效果见表2。从实施例各组与模型组比较可知,小鼠的自愈效果差,有利于观察制剂对胃溃疡的治疗效果。与模型组比较,实施例1～5组均有较好的胃溃疡治疗效果。
测试例17
实施例1组、实施例2组、实施例3组、实施例4组、实施例5组。对照组自由饮食进水,除对照组外,其他组分别经胃镜在胃壁黏膜表面造成2个创面,并进行充分的止血。即水对照组(等体积水灌胃)、实施例1组(0.3ml/20g)、实施例2组(0.3ml/20g)、实施例3组(0.3ml/20g)、实施例4组(0.3ml/20g)、实施例5组(0.3ml/20g)。每日灌胃一次连续14天,第14天灌胃后开始禁食48小时,不禁水。统一拉颈处死小鼠,剖腹,结扎喷门和幽门,向胃内注射1％甲醛溶液1.0ml,取胃置于同一浓度的甲醛溶液内固定30分钟,胃取出后沿胃大弯剪开,将胃外反倒去内容物,用水轻轻冲洗去除胃内残渣。观察腺胃部黏膜溃疡的程度,按照小鼠的溃疡面程度分为痊愈、有效、无效。
对小鼠盐酸实验性胃溃疡的保护作用(实施例21～25所得制剂)。小鼠70,体重18-20克,雌雄各 半。随机分成7组,每组10只,即对照组、模型组、实施
痊愈:小鼠溃疡面完全消失；
有效:小鼠溃疡面有良好改善,且溃疡面患处完好1/2以上；
无效:小鼠溃疡面无明显改善,且溃疡面患处未达到1/2以上完好。实验结果如下；
表3对小鼠盐酸实验性胃溃疡的实验结果
组别 痊愈数 有效数 无效数
对照组 -- -- --
模型组 0 1 9
实施例1组 8 2 0
实施例2组 7 2 1
实施例3组 6 3 1
实施例4组 7 1 2
实施例5组 7 2 1
注:所有变化由相对于对照组的变化确定。
从实施例各组与模型组比较可知,小鼠的自愈效果差,有利于观察制剂对胃溃疡的治疗效果。如上表所示，实施例21～25能明显促进的胃黏膜损伤性溃疡愈合,对胃溃疡有明显的治疗效果。
测试例17
对大鼠冰乙酸实验性胃溃疡的保护作用(实施例21～25所得制剂)。
将体重200～250gSPF级雄性大鼠80只,分成8组每组10只,随机分为正常对照组、模型对照组、自愈组、实施例1组(0.3ml/20g)、实施例2组(0.3ml/20g)、实施例3组(0.3ml/20g)、实施例4组(0.3ml/20g)、实施例5组(0.3ml/20g)。大鼠饮食不限,正常对照组自由饮食进水,17天后处死取材；除正常对照组外,其余各组大鼠均适应性饲养3天后进行造模。术前禁食不禁水24h,10％水合氯醛按3.5ml/kg剂量腹腔注射麻醉大鼠,将大鼠腹部备皮后仰卧固定在解剖办上,常规2％碘酊、75％乙醇消毒大鼠腹部皮肤,在大鼠剑突下腹部正中作一切口,长2cm左右。沿腹白线剪开腹壁组织、打开腹腔,以无齿镊拨动鼠肠找到鼠胃,把腺胃部拉出腹外,用直径5mm 的滤纸片浸渍100％冰乙酸在胃窦部浆膜面同一部位(避开血管)贴敷30s*2次,用干棉球吸除残余的冰乙酸,再用蘸有生理盐水的棉签轻轻擦洗冰乙酸贴敷部位,然后将胃送入腹腔。用1号丝线将腹膜和腹壁各层组织一起间断缝合,再用7号丝线间断缝合皮肤,然后用2％碘酊消毒伤口及周围皮肤。模型对照组于手术后第4天处死取材；自愈组造模后自由饮食进水,实施例五组于手术后第4天开始灌胃,灌胃10天后一起处死取材。然后将福尔马林溶液(1％)注入胃内。将鼠胃在10％福尔马林溶液中固定10分钟,沿胃的弯曲切开,用自来水冲洗。用卡尺测量溃疡面最大横径的d1、最大纵径d2,溃疡面积按公式S＝π*d1*d2*1/4计算。溃疡面完全消失计为溃疡愈合。表4对大鼠冰乙酸实验性胃溃疡的实验结果
组别 溃疡面积(mm2) 溃疡愈合只数
正常对照组 -- --
模型对照组 19.23 0
自愈组 9.46 1
实施例1组 3.72 8
实施例2组 4.05 7
实施例3组 3.82 6
实施例4组 3.74 8
实施例5组 3.40 9
从模型对照组与自愈组的比较,可见本造模方法成功率高,小鼠的自愈效果差,有利于观察制剂对胃溃疡的治疗效果。实施例1～5组,均有明显的治疗效果。
测试例18对狗实验性胃溃疡的保护作用。
狗64只,体重9-12千克,雌雄不限。随机分为8组,每组8只:对照组、模型组、对比例1组、对比例2组、对比例3组、实施例1组、实施例3组、实施例5组。(同之前的测试例一样，实施例1组对应实施例21的制剂，实施例3组对应实施例23的制剂，实施例5组对应实施例25的制剂)。
除对照组外,其余各组麻醉后经胃镜在胃壁粘膜表面造成2个1cmX1cm的创面,并进行充分的止血。术后第3和第6天,实施例各组分别通过胃窥镜喷涂0.3ml制剂。术后第3天开始, 对比例1组每日灌胃0.3ml实施例1制剂,对比例2组每日灌胃0.3ml实施例2制剂,对比例3组每日灌胃0.3ml实施例3制剂,连续给药7天。
第5天、第10天、第14天观察胃部粘膜损伤修复的程度。按照小鼠的溃疡面程度分为痊愈、有效、无效。
痊愈:小鼠溃疡面完全消失；
有效:小鼠溃疡面有良好改善,且溃疡面患处完好1/2以上；
无效:小鼠溃疡面无明显改善,且溃疡面患处未达到1/2以上完好。
表5对狗实验性胃溃疡的实验结果
Figure PCTCN2020078733-appb-000009
注:所有变化由相对于对照组的变化确定。
通过和模型组的对比,本发明制造的制剂对胃溃疡有明显的治疗作用。通过实施例和对比例的对比,本发明制剂通过胃窥镜喷涂,治疗效果显著提升。
第六部分促进消化道黏膜上皮细胞增殖和迁移的成分
实施例26
1.海藻酸钠 11克
2.柠檬酸钠 4克
3.β-葡聚糖 0.2g
4.氯化钙 1.5克
5.多聚赖氨酸 10g
以上成份溶解在0.01mol采用1mol氢氧化钠调pH9.0磷酸盐溶液500ml中溶解，每瓶5ml包装,高温蒸汽灭菌。固化液采用葡萄糖酸钙配制成2％的水溶液，每瓶5ml包装,高温蒸汽灭菌。
实施例27
1.海藻酸钾 10克
2.柠檬酸钠 4克
3.乳酸钙 2.8克
4.β-葡聚糖 0.15g
5.多聚赖氨酸 10g
以上成份溶解在0.01mol采用1mol氢氧化钠调pH9.0磷酸盐溶液600ml中溶解，每瓶5ml包装,高温蒸汽灭菌。固化液采用乳酸钙配制成5％的水溶液，每瓶5ml包装,高温蒸汽灭菌。
实施例28
1.泊洛沙姆407 18克
2.泊洛沙姆188 0.5克
3.羟丙基甲基纤维素 0.5克
4.海藻酸钠 0.3克
5.β-葡聚糖 0.15g
以上成份在4℃溶解在60ml纯水中，溶解完全呈透明液体，然后定容到100ml，每瓶5ml包装。高温蒸汽灭菌。低温保存。
固化液采用氯化钙配制成1.1％的水溶液，每瓶5ml包装,高温蒸汽灭菌。
实施例29
1.聚N-异丙基丙烯酰胺 10克
2.泊洛沙姆188 0.5克
3.胶原蛋白 2克
4.去乙酰结冷胶 0.2克
5.β-葡聚糖 0.15g
以上成份在4℃溶解在60ml生理盐水中，溶解完全呈透明液体，然后定容到100ml，每瓶5ml包装,高温蒸汽灭菌。低温保存。
固化液采用乳酸钙配制成5％的水溶液，每瓶5ml包装,高温蒸汽灭菌。
实施例30
1.海藻酸钠 11克
2.柠檬酸钠 4克
3.氯化钙 1.5克
4.多聚赖氨酸 10g
5.β-葡聚糖 0.15g
以上成份溶解在采用1mol氢氧化钠调pH8.0生理盐溶液1500ml中溶解，每瓶15ml包装,高温蒸汽灭菌。
实施例31
药物组合物原料:
Figure PCTCN2020078733-appb-000010
以上成份溶解在蒸馏水中,配置成质量浓度为3％,每瓶2.5ml包装,灭菌保存。
测试例19保护胶对胃粘膜上皮细胞增殖和迁移效果
1样品制备
11按照产品说明书进行DMEM培养液(高糖)的配制，补充10μg/mL牛胰岛素，50mg/mL庆大霉素，10％胎牛血清。实施例按照0.2克样品加入DMEM培养液在37℃浸提24小时，取液体部分用0.22μm微孔滤膜过滤除菌，4℃冰箱避光保存备用。
1.2细胞培养人胃粘膜上皮细胞，置37℃、饱和湿度、5％CO 2无菌培养箱培养，每2-3d换液1次，每3-4d传代，使细胞浓度维持70％-80％的融合度。
2对胃黏膜上皮细胞增值的影响
2.1对人胃粘膜上皮细胞GES-1增殖的影响：取对数生长期细胞，将其浓度调整为2.0×10 4个/mL，接种于96孔板，每孔100μL，置37℃、饱和湿度、5％CO 2无菌培养箱培养24h后更换培养液，实验组分别加入实施例浸提液各200μL，空白组为10％FBS的DMEM，溶剂空白组为相对应的培养液，每组设6个平行复孔，培养24h、48h后，加MTS溶液20μL，继续培养4h，测吸光度(A492)，计算细胞增殖率。
细胞增殖率＝(A实验-A溶剂空白)/(A对照组-A溶剂空白)X100％
3对胃黏膜上皮细胞迁移的影响
3.1对人胃粘膜上皮细胞GES-1迁移的影响细胞迁移的造模过程。
具体如下：对数生长期的人胃黏膜上皮细胞，以每孔6X10 5个细胞接种于24孔板，每孔400μL，置37℃，5％CO 2培养箱中培养24h，待细胞间形成较紧密的连接成单层后，用直径为1.5mm的毛细管在培养皿中央垂直划一道痕迹，且在显微镜下可同时看到创面两侧。划伤后吸出原培养液，每孔用200μL磷酸缓冲溶液(PBS)洗2次以除净未贴壁细胞，然后各孔加入本部分各实施例浸提液400μL，对照组(CK)仅加入DMEM培养液400μL，应用软件Image pro plus 6.0分别测量0、6、12、24h的划痕创面愈合情况。
细胞迁移率＝0h创面面积-Th创面面积)/(0h创面面积)X100％
4结果
4.1实施例对胃黏膜上皮细胞促进增殖的结果
试验结果表明实施例能明显促进胃黏膜细胞的增殖，增殖的速度是空白的1.5倍以上。
Figure PCTCN2020078733-appb-000011
4.2实施例对胃黏膜上皮细胞促进迁移的结果
试验结果表明实施例能明显促进胃黏膜细胞的迁移，迁移的速度是空白2倍以上。
Figure PCTCN2020078733-appb-000012
研究结果表明实施例能明显促进胃黏膜上皮细胞增殖和迁移，因此通过加速黏膜细胞增殖和向创面的迁移，促进消化道黏膜的愈合。
本文中所描述的具体实施例仅仅是对本发明精神作举例说明。本发明所属技术领域的技术人员可以对所描述的具体实施例做各种各样的修改或补充或采用类似的方式替代，但并不会偏离本发明的精神。

Claims (28)

  1. 消化道黏膜保护胶，其特征在于，所述消化道黏膜保护胶包括保护层和生物粘合剂；
    其中保护层的有效成分为成胶物质，水溶性钙盐和钙离子络合剂；
    成胶物质为果胶、海藻酸盐、去乙酰结冷胶中的一种或多种；
    生物粘合剂为羧甲基纤维素、羧甲基壳聚糖、葡聚糖、卡波姆、乙基纤维素、羟丙基纤维素、羟丙基甲基纤维素、白芨多糖、多聚赖氨酸、贻贝粘性蛋白、胶原蛋白、聚多巴胺、透明质酸钠、壳聚糖季铵盐中的一种或多种；
    所述消化道黏膜保护胶的pH值不小于7。
  2. 根据权利要求1所述的消化道黏膜保护胶，其特征在于，钙离子络合剂为柠檬酸、柠檬酸盐、EDTA或EDTA钠盐中的一种或几种。
  3. 根据权利要求1所述的消化道黏膜保护胶，其特征在于，所述果胶为低甲氧基果胶和酰胺化低甲氧基果胶中的一种或两种。
  4. 根据权利要求1所述的消化道黏膜保护胶，其特征在于，所述海藻酸盐为海藻酸钠或海藻酸钾中的一种或两种。
  5. 根据权利要求1所述的消化道黏膜保护胶，其特征在于，钙盐溶液中钙盐的质量浓度为0.25％-5％。
  6. 根据权利要求1所述的消化道黏膜保护胶，其特征在于，成胶物质与钙离子络合剂重量比为3-6:1。
  7. 根据权利要求1所述的消化道黏膜保护胶，其特征在于，所述生物粘合剂为多聚赖氨酸，成胶物质与多聚赖氨酸的重量比为3：1-3。
  8. 根据权利要求7所述的消化道黏膜保护胶，其特征在于，多聚赖氨酸在钙盐溶液中的重量百分比为0.8-3％。
  9. 根据权利要求1所述的消化道黏膜保护胶，其特征在于，所述生物粘合剂为含有贻贝粘蛋白的水溶液。
  10. 根据权利要求9所述的消化道黏膜保护胶，其特征在于，贻贝粘蛋白为生物粘合剂的5-25wt％。
  11. 根据权利要求9所述的消化道黏膜保护胶，其特征在于，保护层组分中还 含有0.3-5wt％的增塑剂。
  12. 根据权利要求11所述的消化道黏膜保护胶，其特征在于，增塑剂为甘油、聚乙二醇、聚乙烯醇、甘露醇和山梨醇中的一种或几种。
  13. 根据权利要求1所述的消化道黏膜保护胶，其特征在于，所述消化道黏膜保护胶还包括温度敏感性生物材料，凝胶化温度调节剂。
  14. 根据权利要求13所述的消化道黏膜保护胶，其特征在于，
    温度敏感性生物材料：凝胶化温度调节剂：生物粘附剂：成胶物质的重量百分比为为1-30：0.5-5：0.3-5：0.2-1。
  15. 根据权利要求13所述的消化道黏膜保护胶，其特征在于，
    所述温度敏感性生物材料为聚N-异丙基丙烯酰胺及其衍生物、羟丁基壳聚糖及其衍生物、泊洛沙姆407、聚乳酸/羟基乙酸/聚乙二醇共聚物、聚乙二醇单甲醚-(癸二酸-D,L-乳酸)聚酯酸酐-聚乙二醇单甲醚三嵌段共聚物、纤维素及其衍生物和聚乙二醇/聚己内酯嵌段共聚物中的一种或多种。
  16. 根据权利要求13所述的消化道黏膜保护胶，其特征在于，
    所述的凝胶化温度调节剂为泊洛沙姆188、聚山梨醇酯、聚乙二醇中的一种或多种。
  17. 根据权利要求1所述的消化道黏膜保护胶，其特征在于，所述保护胶还包括固定液，固定液为水溶性钙溶液。
  18. 根据权利要求17所述的消化道黏膜保护胶，其特征在于，所述水溶性钙为氯化钙、乳酸钙、葡萄糖酸钙中的一种或多种。
  19. 根据权利要求17所述的消化道黏膜保护胶，其特征在于，水溶性钙溶液浓度0.3％-5％。
  20. 根据权利要求11所述的消化道黏膜保护胶，其特征在于，所述保护胶还包括治疗溃疡、肿瘤的有效药物量的药物化合物或组合物。
  21. 根据权利要求1所述的消化道黏膜保护胶，其特征在于，所述保护胶还包括胃酸抑制剂、抗幽门杆菌制剂。
  22. 根据权利要求1所述的消化道黏膜保护胶，其特征在于，胃酸抑制剂为质子泵抑制剂或H2受体拮抗剂。
  23. 根据权利要求1所述的消化道黏膜保护胶，其特征在于，抗幽门杆菌制剂 由铋剂、甲硝唑或替硝唑、阿莫西林、克拉霉素、四环素、呋喃唑酮中的一种或多种组成。
  24. 根据权利要求1所述的消化道黏膜保护胶，其特征在于，胃酸抑制剂、抗幽门杆菌制剂和保护胶其他组分的重量比为0.001-0.05:0.001-1:1-20。
  25. 根据权利要求1所述的消化道黏膜保护胶，其特征在于，所述保护胶还包括甘草多糖、β-葡聚糖、党参多糖、索拉胶、杏鲍菇葡聚糖、燕麦葡聚糖、灰树花多糖、谷物葡聚糖、酵母葡聚糖、灵芝多糖中的一种或多种。
  26. 药物组合物，其特征在于，所述药物组合物含有如权利要求1-25任意一项所述的消化道黏膜保护胶和其他具有治疗效果的药物。
  27. 消化道隆起注射剂，其特征在于，所述注射剂包含如权利要求1-25任意一项所述的消化道黏膜保护胶。
  28. 如权利要求1-25任意一项所述的消化道黏膜保护胶在植被治疗消化性溃疡、应激性溃疡、溃疡性结肠炎、胃炎、胃黏膜切除和剥离术后的创面、食道粘膜剥离术和肠道粘膜剥离术后的创面、胃泌素瘤和胃癌药物中的应用。
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