WO2017028775A1 - 贻贝粘蛋白用于粘膜的防护 - Google Patents

贻贝粘蛋白用于粘膜的防护 Download PDF

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Publication number
WO2017028775A1
WO2017028775A1 PCT/CN2016/095362 CN2016095362W WO2017028775A1 WO 2017028775 A1 WO2017028775 A1 WO 2017028775A1 CN 2016095362 W CN2016095362 W CN 2016095362W WO 2017028775 A1 WO2017028775 A1 WO 2017028775A1
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Prior art keywords
mussel mucin
mussel
mucin
rats
protecting
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PCT/CN2016/095362
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English (en)
French (fr)
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高敏
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江阴市本特塞缪森生命科学研究院有限公司
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Publication of WO2017028775A1 publication Critical patent/WO2017028775A1/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
    • A61L15/00—Chemical aspects of, or use of materials for, bandages, dressings or absorbent pads
    • A61L15/16—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons
    • A61L15/22—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons containing macromolecular materials
    • A61L15/32—Proteins, polypeptides; Degradation products or derivatives thereof, e.g. albumin, collagen, fibrin, gelatin
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2/00—Peptides of undefined number of amino acids; Derivatives thereof

Definitions

  • the invention generally relates to the fields of daily chemical products, cosmetics, medical devices, medicines, disinfection products, health care products, and the like, and relates to the application of mussel mucin or its preparation in protective mucosa.
  • the mucous membrane is the inner wall of various organs such as digestion, respiration, excretion, and reproduction in the body, and the surface of the mucus is kept moist by mucus, specifically including the moist lining of the oral cavity, the nasal cavity, the intestine, the vagina, the intestine, and the like.
  • the skin and mucous membrane protect the human body, so that the human body forms a closed system to the outside world.
  • the skin and mucous membranes When harmful substances invade the human body, the skin and mucous membranes first block the external pathogenic factors outside the body. Therefore, skin and mucous membranes are the first line of defense against infection.
  • the mucosa is susceptible to stimulation by physical factors, chemical factors, bacteria or their toxins.
  • Physical factors include ultraviolet rays, radiation (such as electromagnetic, radiation, computer radiation, etc.), over-cooling, over-scalding, seawater, and environmental pollution.
  • Chemical factors include chemical agents, irritating drugs, foods, and the like.
  • UVB Ultraviolet light
  • Radiation such as ionizing radiation, including radiotherapy, has a certain therapeutic effect on malignant tumors, but a certain degree of mucosal reaction occurs during treatment, such as erythema, burning and itching, congestion, edema, exudation, erosion, ulceration, Capillary expansion, lymphatic drainage disorders, etc.
  • the scald is caused by inhalation of high heat steam, accidental drinking of boiling water, head and neck burns caused by inhalation of flame or dry hot air, redness of the mucosa, blister tissue necrosis, ulceration, ulcers and the like.
  • Frostbite is a local damage of mucous membrane caused by cold and dampness. It is cold, pale, hard, numbness or loss, redness and congestion, heat, itching, burning pain, blisters, edema, severe pain and tissue necrosis.
  • Salt in seawater and chlorine in swimming pools can damage the skin and mucous membranes, causing oral mucosa Inflammation, inflammation, rhinitis, pharyngitis and other inflammatory reactions can also make the hair dry.
  • the main pollutants in the air are sulfur dioxide, nitrogen oxides, particulate pollutants, and acid rain.
  • Nitrogen-containing contaminants such as sulfur dioxide (SO 2 ), nitrous oxide (N 2 O), nitric oxide (NO), nitrogen dioxide (NO 2 ), and dinitrogen tetroxide (N 2 O 3 ) It is very irritating.
  • the amount of particulate contaminants is large and the composition is complex. It can be a carrier of toxic substances or other pollutants, which may cause problems such as mucosal inflammation.
  • Smoking, drinking, strong tea, coffee, irritating drugs, food, chemical reagents, etc. can cause damage to the digestive tract and respiratory mucosa.
  • Mussel adhesive protein also known as Mytilus edulis foot protein (Mefp)
  • Mefp Mytilus edulis foot protein
  • Mytilus coruscus A special protein secreted by Perna viridis. Mussels are usually attached in groups to the reefs on the coast or to the bottom of the ship, and have the ability to withstand wave impacts in the offshore. In fact, mussels can be attached extremely strongly to the substrate of any material, such as metal, wood, glass, and the like. The main reason for the above characteristics of mussels is that they can form and store this special mucin in the girth of the foot. The mussels release the mucin through the foot silk to a solid surface such as rock to form a water-resistant combination. Fix yourself.
  • Mussel mucin has two structural features: (1) containing lysine, which has a high loading of positive charge; (2) containing 3,4 dihydroxyphenylalanine (DOPA, dopa). The cells and tissues of the human body are negatively charged.
  • Mussel mucin plays a protective and therapeutic role by tightly binding cells and tissues through the electrostatic interaction between its own positive charge and the negative charge of cells and tissues.
  • dopa oxidation produces ortho-dioxins, which can be cross-linked with unoxidized dopa to form a membrane or a network scaffold, which promotes the protein to adhere more closely and firmly to the surface of the human body, thereby protecting.
  • Mussel mucin is a macromolecular protein that is completely degraded in the human body for about 3-10 days. Its ability to attach to cell tissues is excellent, so that mussel mucin can be stabilized locally and continue to function.
  • mussel mucin has the above characteristics, its current application field is very limited.
  • Commercial mussel mucin products are Cell-Tak from BD Biosciences, MAP Trix from Kollodis, Korea, and Hydrogel from Biopolymer, Sweden.
  • Mussel mucin The solution state is used as it is, or it is stored as a lyophilized powder formulation and dissolved before use. Their main fields of application are limited to microscopic cell adhesion and tissue adhesives. Mussel mucin has also been reported for use in the repair of fetal membranes, as a coating against seawater corrosion, and as a drug-loaded stent for the heart.
  • Mussel mucin used herein refers to Mytilus edulis Linnaeus, Mytilus coruscus or Perna viridis from the Mytilidae bivalve mollusc. 11 subclasses of mussel mucin, currently known as purified from marine mussels: mefp1, mefp-2, mefp-3, mefp-4, mefp-5, mefp-6, collagen pre-COL- A mixture of one or more of P, pre-COL-D, pre-COL-NG, foot silk matrix proteins PTMP and DTMP.
  • the mussel mucin used herein may have a pH of 1.0 to 7.0 in an aqueous solution, and particularly may be in the range of pH 3.0 to 6.5 to make the therapeutic effect better.
  • the mussel mucin used herein can be obtained by the following preparation method, for example, "A method for separating and purifying mussel mucin using mixed adsorption chromatography", Chinese Patent No. ZL200710179491.0, “China Patent No. ZL200710179492.5” A method for purifying mussel mucin using carboxymethyl ion exchange chromatography", "A method for separating and purifying mussel mucin using salting out and dialysis", Chinese Patent No. ZL200910087567.6, and the like.
  • the mussel mucin used herein may be in the form of a solution or a lyophilized powder, in particular, the concentration of mussel mucin in the product may be 0.1-15.0 mg/ml, and when the concentration is too low, the effect of mussel mucin Not large, when the concentration is too high, the cytotoxicity may be caused by the high positive charge, which is not conducive to play a protective role.
  • the mussel mucin used herein can also be prepared as a liquid agent by combining it with an excipient.
  • An exemplary mussel mucin liquid preparation is prepared by dissolving or diluting mussel mucin solution mother liquor or lyophilized powder to a certain concentration or pH, and the solution for dissolving or diluting may be water, physiological saline, phosphate solution, vinegar. Acid solution, borate solution, and the like.
  • the pH of mussel mucin in the final product may be pH 1.0-7.0, and in particular, the therapeutic effect may be better in the range of pH 3.0-6.5.
  • the mussel mucin used herein can also be prepared as a gelling agent in combination with an excipient.
  • An exemplary mussel mucin gel is prepared by mixing a mussel mucin solution or a lyophilized powder with a gel matrix material, which may be selected from the group consisting of cellulose derivatives, carbomers, and seaweeds. Acid salt, western yellow One of silicone, gelatin, pectin, carrageenan, gellan gum, starch, xanthan gum, cationic guar gum, agar, non-cellulosic polysaccharide, ethylene polymer, acrylic resin, polyvinyl alcohol or carboxyvinyl Or any combination thereof.
  • the mussel mucin used herein can also be made into a foaming agent.
  • An exemplary mussel mucin foaming agent is prepared by mixing a mussel mucin solution or a lyophilized powder with a foaming agent matrix, which may include hydroxypropylmethylcellulose, gelatin, and polyethylene.
  • a foaming agent matrix which may include hydroxypropylmethylcellulose, gelatin, and polyethylene.
  • the defoaming time of the foaming agent is long, and the action time is prolonged, so that the mussel mucin is more effective.
  • the mussel mucin used herein can be used as a main raw material to prepare a medicine using a pharmaceutically acceptable carrier.
  • the drug may be a liquid agent, a gel, or a foaming agent.
  • the medicine can be administered orally, by sublingual (sublingual), perfusion (rectal administration), dripping (eye), spraying (mouth, nose), inhalation (mouth, nose), spraying (mouth, nose, ear, cervix) Etc., laparoscopic (uterine cavity, abdominal cavity, etc.), targeted local sustained release, targeted administration, and can be administered at low temperature or in a heated manner.
  • the mussel mucin used herein can be used as a main raw material to prepare a medical device.
  • the term medical device as used herein refers to materials that are used directly or indirectly to the human body and other similar or related items.
  • the medical device may be a liquid agent, a gel, or a foaming agent.
  • the medical device can be administered orally, by sublingual (sublingual), perfusion (rectal administration), dripping (eye), spraying (mouth, nose), inhalation (mouth, nose), spraying (mouth, nose, ear, Cervical, etc., laparoscopic (uterine cavity, abdominal cavity, etc.), targeted local sustained release, targeted administration, and can be administered at low temperature or in a heated manner.
  • the mussel mucin used herein can be used as a main raw material to prepare a cosmetic using an auxiliary material which is acceptable in the field of cosmetics.
  • the cosmetic may be a liquid, a gel, or a foaming agent.
  • the cosmetic can be administered orally, by sublingual (sublingual), perfusion (rectal administration), dripping (eye), spraying (mouth, nose), inhalation (mouth, nose), spraying (mouth, nose, ear, cervix) Etc., laparoscopic (uterine cavity, abdominal cavity, etc.), targeted local sustained release, targeted administration, and can be administered at low temperature or in a heated manner.
  • the mussel mucin used herein can be used as a main raw material to prepare a disinfecting product using an excipient which is acceptable in the field of disinfecting products.
  • the term disinfecting product as used herein refers to a disinfectant, a disinfecting device, a sanitary article, and a disposable medical article that chemically, physically, or biologically kill or eliminate pathogenic microorganisms in the environment.
  • the disinfecting product may be a liquid agent, a gelling agent, or a foaming agent.
  • the disinfecting product can be administered orally, by sublingual (sublingual), perfusion (rectal administration), dripping (eye), spraying (mouth, nose), inhalation (mouth, nose), spraying (mouth, nose, ear, Cervical, etc., laparoscopic (uterine cavity, abdominal cavity, etc.), targeted local sustained release, targeted administration, and can be administered at low temperature or in a heated manner.
  • the mussel mucin used herein can be used as a main raw material, and a health care product or food can be prepared by using an excipient which is acceptable in the field of health care or food.
  • the health supplement or food may be a liquid, a gel, or a foam.
  • the health care product or food can be administered orally, by sublingual (sublingual), perfusion (rectal administration), dripping (eye), spraying (mouth, nose), inhalation (mouth, nose), spraying (mouth, nose, Ear, cervix, etc.), laparoscopic (uterine cavity, abdominal cavity, etc.), targeted local sustained release, targeted administration, and can be administered at low temperature or heating.
  • the mussel mucin used herein can be used as a main raw material to prepare a daily chemical product using an auxiliary material acceptable in the field of daily chemical products.
  • the term daily chemical product as used herein refers to technical chemicals for daily use, including shampoos, shower gels, and the like.
  • the daily chemical product may be a liquid agent, a gel agent, or a foaming agent.
  • the daily chemical product can be administered orally, by sublingual (sublingual), perfusion (rectal administration), dripping (eye), spraying (mouth, nose), inhalation (mouth, nose), spraying (mouth, nose, ear) , cervix, etc.), laparoscopic (uterine cavity, abdominal cavity, etc.), targeted local sustained release, targeted administration, and can be administered at low temperature or heating.
  • Another object of the invention is to provide a mussel mucin product for use in mucosal protection.
  • mucosal protection refers to the protection of mucous membranes including the oral cavity, nasal cavity, intestinal tract, vagina, intestines and the like from physical and chemical damage by protective measures.
  • the present invention provides the use of mussel mucin or a preparation thereof for protecting a mucous membrane from ultraviolet damage or the like.
  • the present invention provides the use of mussel mucin or a preparation thereof for protecting a mucous membrane from radiation (electromagnetic, radiation, computer radiation, etc.).
  • the present invention provides the use of mussel mucin or a preparation thereof for protecting a mucous membrane from a burn caused by a high temperature object or the like.
  • the present invention provides the use of mussel mucin or a preparation thereof for protecting a mucosa from frostbite caused by a low temperature object or the like.
  • the present invention provides the use of mussel mucin or a preparation thereof for protecting a mucous membrane from seawater damage; and for protecting the mucosa from environmental pollution or the like.
  • the present invention provides the use of mussel mucin or a preparation thereof for protecting a mucous membrane from damage from smoking, drinking, tea, coffee, irritating drugs, foods, chemical agents and the like.
  • the present invention provides a mussel mucin-containing liquid agent for protecting mucous membranes from smoking, drinking, tea, coffee, irritating drugs, foods, chemicals, and the like.
  • the present invention provides a mussel mucin-containing gelling agent for protecting a mucous membrane from damage from smoking, drinking, tea, coffee, irritating drugs, foods, chemicals, and the like.
  • the present invention provides a mussel mucin-containing foaming agent for protecting a mucous membrane from damage from smoking, drinking, tea, coffee, irritating drugs, foods, chemicals, and the like.
  • the present invention provides a method for preparing a mussel mucin-containing liquid agent for protecting a mucous membrane from damage from smoking, drinking, tea, coffee, irritating drugs, foods, chemicals, and the like. .
  • the present invention provides a mussel mucin-containing gel preparation for protecting a mucous membrane from smoking, drinking, tea, coffee, irritating drugs, foods, chemicals, and the like. method.
  • the present invention provides a mussel-containing mucin-containing foaming agent for the protection of mucosa from smoking, drinking, tea, coffee, irritating drugs, foods, chemicals, and the like.
  • the mucosal inflammation may comprise any one as described herein above.
  • mussel mucin application according to embodiment 1, wherein the mussel mucin may be from a subclass: mefp1, mefp-2, mefp-3, mefp-4, mefp-5, mefp-6, collagen pre-COL-P, A mixture of one or more of pre-COL-D, pre-COL-NG, foot silk matrix protein PTMP and DTMP.
  • the mussel mucin application according to embodiment 1, wherein the mussel mucin concentration may be from 0.1 to 15.0 mg/ml.
  • Mussel mucin application according to embodiment 1, wherein the mussel mucin in the final product may be in the range of pH 1.0-7.0, in particular in the range of pH 3.0-6.5.
  • composition can be administered orally, by sublingual (sublingual), perfusion (rectal administration), instillation (eye), spraying (mouth, nose), inhalation. (mouth, nose), spraying (mouth, nose, ear, cervix, etc.), laparoscopic (uterine cavity, abdominal cavity, etc.), targeted local sustained release, targeted drug delivery.
  • a gelling agent for mucosal protection comprising mussel mucin as an active ingredient.
  • a liquid agent for mucosal protection comprising mussel mucin as an active ingredient.
  • a foaming agent for mucosal protection comprising mussel mucin as an active ingredient.
  • the protective mucosa may include protective mucous membranes from ultraviolet rays, radiation, high temperature objects, low temperature objects, sea water, environmental pollution, chemical agent stimulation, and irritating drugs, food damage.
  • a method of preparing a gel for mucosal protection which may comprise: 1) obtaining or preparing Mussel mucin solution or lyophilized powder, 2) obtaining or preparing a gel matrix, 3) mixing the mussel mucin solution of step 1) and the gel matrix of step 2), and 4) adjusting the pH to weakly acidic, obtaining Gelling agent.
  • a method of preparing a liquid agent for mucosal protection which may comprise: obtaining or preparing a mussel mucin solution or a lyophilized powder, diluting with a pharmaceutically acceptable diluent, and adjusting the pH to a weakly acidic state to obtain a liquid Agent.
  • a method of preparing a foaming agent for mucosal protection which comprises: obtaining or preparing a mussel mucin solution or a lyophilized powder, mixing with a pharmaceutically acceptable foaming agent base, and adjusting the pH to a weakly acidic state. , obtaining a foaming agent.
  • Example 1 Application of mussel mucin hydrogel drug in gastric mucosal protection.
  • Healthy male rats were tested at a dose of 180-200 g each for a total of 20 rats, and 10 rats in the control group and the test group.
  • the control group was intragastrically administered with absolute ethanol, 1.0 ml each time. After 1 hour, the animals were sacrificed, the whole stomach was exposed, the pylorus was ligated, and 10% formaldehyde solution was fixed for 20 minutes. Then the skin was cut along the stomach, the stomach contents were washed, the gastric mucosa was removed, and the bleeding was measured with a vernier caliper under a stereo dissection microscope. The length and width of the point or bleed zone.
  • the test group first took the above-mentioned mussel mucin hydrogel drug 2.0 ml, and after 1 hour, the same method as the control group was used to administer the stomach with absolute ethanol, and the bleeding point was measured.
  • the control group and the test group evaluated the degree of gastric mucosal damage by the degree of gastric mucosal damage (see Table 1 for evaluation methods) and the incidence of injury. It can be seen from the degree of gastric mucosal injury and the incidence of injury (see Table 2) that The use of mussel mucin hydrogel drugs can reduce the incidence of gastric mucosal damage and reduce the degree of damage.
  • Table 1 Scoring criteria for acute alcohol injury observation
  • Incidence rate (%) number of rats with bleeding or ulceration / total rat x 100%
  • Example 2 Application of mussel mucin liquid health care product in gastric mucosal protection.
  • a mussel mucin solution was prepared and formulated into an aqueous solution having a concentration of 5.0 mg/ml with 0.5% acetic acid to obtain a mussel mucin liquid health product.
  • Healthy male rats were tested at a dose of 180-200 g each for a total of 20 rats, and 10 rats in the control group and the test group.
  • the control group was intragastrically administered with absolute ethanol, 1.0 ml each time. After 1 hour, the animals were sacrificed, the whole stomach was exposed, the pylorus was ligated, and 10% formaldehyde solution was fixed for 20 minutes. Then the skin was cut along the stomach, the stomach contents were washed, the gastric mucosa was removed, and the bleeding was measured with a vernier caliper under a stereo dissection microscope. The length and width of the point or bleed zone.
  • the test group first took 3.0 ml of mussel mucin liquid health care product, and after 1 hour, it was orally administered with absolute ethanol according to the same method as the control group, and the bleeding point was measured.
  • the control group and the experimental group evaluated the degree of gastric mucosal damage by the degree of gastric mucosal damage (see Table 3 for evaluation methods) and the incidence of injury. From the degree of gastric mucosal damage and the incidence of injury (see Table 4), mussel mucin was used. Hydrogel drugs can reduce the incidence of gastric mucosal damage and reduce the degree of damage.
  • Incidence rate (%) number of rats with bleeding or ulceration / total rat x 100%
  • Example 3 Application of mussel mucin hydrogel medical device in the protection of high temperature inhalation injury.
  • a mussel mucin solution having a concentration of 5.0 mg/ml was prepared and diluted with physiological saline to prepare a mussel mucin liquid medical device, wherein the mussel mucin concentration was 2.5 mg/ml.
  • mice Twenty white rats weighing 180g-200g and 10 rats were used as the experimental group.
  • the above-mentioned mussel mucin liquid medical device was inhaled continuously for 30 minutes.
  • Another 10 samples were used as the control group, and no other treatment was performed before the test.
  • the rats were fixed on a rat plate and placed under high temperature steam for 30 sec to form an inhalation injury.
  • the nasal cavity, mouth, pharynx, and throat mucosa of the inhalation injury were weighed and weighed, then placed in a 60 ° C oven for 48 hours, and the dry weight was weighed.
  • the mass of the mucosa after drying was divided by the mass of the mucosa before drying, and the ratio obtained was used to evaluate the degree of edema caused by the damage. The higher the ratio, the lighter the edema.
  • the dry weight/wet weight of the test sites of the 10 test rats was 23.4% ⁇ 1.8%; the dry weight/wet weight of the test sites of the 10 control rats was 12.9% ⁇ 1.2%.
  • the edema of the test site coated with the mussel mucin liquid medical device was significantly lighter than that of the control group. It is indicated that the mussel mucin product has the function of protecting the mucous membrane from high temperature gas.
  • Example 4 Application of mussel mucin liquid medicine in chemical stimulation protection.
  • a mussel mucin solution having a concentration of 10.0 mg/ml was prepared and diluted with physiological saline to prepare a mussel mucin liquid medicine, wherein the mussel mucin concentration was 5.0 mg/ml.
  • mice Twenty white rats weighing 180g-200g and 10 rats were used as the experimental group. The above-mentioned mussel mucin liquid medicine was inhaled continuously for 30 minutes. Another 10 samples were used as the control group, and no other treatment was performed before the test. After anesthesia, the rats were fixed on a rat plate and placed under the vapor of chemical reagents for 30 sec to form a chemically irritating lesion.
  • the dry weight/wet weight of the test sites of the 10 test rats was 35.1% ⁇ 2.9%; the dry weight/wet weight of the test sites of the 10 control rats was 19.3% ⁇ 2.6%.
  • the edema of the test site in which the mussel mucin liquid drug was inhaled was significantly lighter than that of the control group. It is indicated that the mussel mucin product has the function of protecting the mucosa from chemical stimulation.
  • Example 5 Application of mussel mucin liquid disinfection product in environmental pollution protection (PM2.5).
  • a mussel mucin solution having a concentration of 10.0 mg/ml was prepared and diluted with physiological saline to prepare a mussel mucin liquid disinfecting product in which the mussel mucin concentration was 2.0 mg/ml.
  • Example 6 Application of mussel mucin hydrogel medical device in ultraviolet protection.
  • Twenty Wistar rats weighing 200g-300g and 10 rats were used as test group.
  • the above-mentioned mussel mucin hydrogel medical device was used to coat the conjunctiva. Another 10 samples were used as the control group.
  • a daylight simulator xenon arc lamp was used as a light source to continuously generate ultraviolet rays having a wavelength of 290 nm to 400 nm, and all the rats were placed under ultraviolet light irradiation once a day for 2 hours.
  • Example 7 Application of mussel mucin liquid cosmetics in radioactive ray protection.
  • a mussel mucin solution having a concentration of 10.0 mg/ml was prepared and diluted with physiological saline to prepare a mussel mucin liquid cosmetic, wherein the mussel mucin concentration was 2 mg/ml.
  • mice Twenty Wistar rats weighing 200 g-300 g were used as the test group, and the above mussel mucoid liquid cosmetics were dropped to the conjunctiva every day; 10 were used as the control group, and no other treatment was performed before the test.
  • the rats were irradiated with radioactive rays at a radiation intensity of 10 mV, and all the rats were placed under irradiation conditions for one time each time for 40 minutes.
  • Example 8 Application of mussel mucin foam medical device in vaginal and cervical mucosal protection
  • the mussel mucin solution was mixed with hydroxypropylmethylcellulose in a ratio of 2:1 by mass ratio, and the acidity and alkalinity was adjusted to pH 5.0 with citric acid to obtain a mussel mucin foam medical device, wherein the mussel mucin concentration was 5 mg/mL.
  • the control group was perfused with phenol cement (5 ml of phenol, 1 g of scutellaria gelatin, 4 ml of glycerin, and 20 ml of distilled water) in the vagina of each mouse at a dose of 0.01 mg/kg body weight once a day for 2 days.
  • the test group was sprayed on the vagina and cervical region using the above-mentioned mussel mucin foam medical device in an amount of 1 ml/mouse. 1 small After the time, the phenol cement was poured in the same treatment method as the control group.
  • the organ index of the experimental group was lower than that of the control group, which proved that the vaginal and mucosal inflammatory response of the experimental group was lower than that of the control group. It can be seen that the mussel mucin product of the present invention has an effect of protecting the vagina and the cervical mucosa.
  • Example 9 Application of mussel mucin gel medical device in vaginal and cervical mucosal protection
  • the mussel mucin solution was mixed with sodium alginate at a mass ratio of 2:1, and the acidity and alkalinity was adjusted to pH 5.0 with citric acid to obtain a mussel mucin gel medical device, wherein the mussel mucin concentration was 5 mg/mL. .
  • the control group was perfused with phenol cement (5 ml of phenol, 1 g of scutellaria gelatin, 4 ml of glycerin, and 20 ml of distilled water) in the vagina of each mouse at a dose of 0.01 mg/kg body weight once a day for 2 days.
  • the test group was sprayed on the vagina and cervical region using the above mussel mucin gel medical device in an amount of 1 ml/mouse. After 1 hour, the phenol cement was poured in the same treatment as the control group.
  • the organ index of the experimental group was lower than that of the control group, which proved that the vaginal and mucosal inflammatory response of the experimental group was lower than that of the control group. It can be seen that the mussel mucin product of the present invention has an effect of protecting the vagina and the cervical mucosa.
  • Example 10 Application of mussel mucin foaming agent in gastric mucosal protection.
  • Healthy male rats were tested at a dose of 180-200 g each for a total of 20 rats, and 10 rats in the control group and the test group.
  • the control group was intragastrically administered with absolute ethanol, 1.0 ml each time. After 1 hour, the animals were sacrificed, the whole stomach was exposed, the pylorus was ligated, and 10% formaldehyde solution was fixed for 20 minutes. Then the skin was cut along the stomach, the stomach contents were washed, the gastric mucosa was removed, and the bleeding was measured with a vernier caliper under a stereo dissection microscope. The length and width of the point or bleed zone.
  • the test group first took 2.0 ml of the above mussel mucin hydrogel drug, and after 1 hour, it was orally administered with absolute ethanol in the same manner as the control group, and the bleeding point was measured.
  • the control group and the test group evaluated the degree of gastric mucosal damage by the degree of gastric mucosal damage (see Table 1 for evaluation methods) and the incidence of injury. From the degree of gastric mucosal damage and the incidence of injury (see Table 8), mussel mucin was used. Foaming agents can reduce the incidence of gastric mucosal damage and reduce the extent of damage.
  • Table 1 Scoring criteria for acute alcohol injury observation
  • Incidence rate (%) number of rats with bleeding or ulceration / total rat x 100%
  • Example 11 Application of mussel mucin liquid medical device in the protection of high temperature inhalation injury.
  • a mussel mucin solution having a concentration of 5.0 mg/ml was prepared and diluted with physiological saline to prepare a mussel mucin liquid medical device, wherein the mussel mucin concentration was 2.5 mg/ml.
  • mice Twenty white rats weighing 180g-200g and 10 rats were used as the experimental group.
  • the above-mentioned mussel mucin liquid medical device was inhaled continuously for 30 minutes.
  • Another 10 samples were used as the control group, and no other treatment was performed before the test.
  • the rats were fixed on a rat plate and placed under high temperature steam for 30 sec to form an inhalation injury.
  • the dry weight/wet weight of the test sites of the 10 test rats was 16.7% ⁇ 1.3%; the dry weight/wet weight of the test sites of the 10 control rats was 12.9% ⁇ 1.2%.
  • the edema of the test site coated with the mussel mucin liquid medical device was significantly lighter than that of the control group. It is indicated that the mussel mucin product has the function of protecting the mucous membrane from high temperature gas.
  • Example 12 Application of mussel mucin foam medical device in the protection of high temperature inhalation injury.
  • mice Twenty white rats weighing 180g-200g and 10 rats were used as the experimental group.
  • the above-mentioned mussel mucin liquid medical device was inhaled continuously for 30 minutes.
  • Another 10 samples were used as the control group, and no other treatment was performed before the test.
  • the rats were fixed on a rat plate and placed under high temperature steam for 30 sec to form an inhalation injury.
  • the nasal cavity, mouth, pharynx, and throat mucosa of the inhalation injury were weighed and weighed, then placed in a 60 ° C oven for 48 hours, and the dry weight was weighed.
  • the mass of the mucosa after drying was divided by the mass of the mucosa before drying, and the ratio obtained was used to evaluate the degree of edema caused by the damage. The higher the ratio, the lighter the edema.
  • the dry weight/wet weight of the test sites of the 10 test rats was 29.1% ⁇ 2.0% on average; the dry weight/wet weight of the test sites of the 10 control rats was 12.9% ⁇ 1.2%.
  • the edema of the test site coated with the mussel mucin liquid medical device was significantly lighter than that of the control group. It is indicated that the mussel mucin product has the function of protecting the mucous membrane from high temperature gas.
  • Example 13 Application of mussel mucin gel drug in chemical stimulation protection.
  • mice Twenty white rats weighing 180g-200g and 10 rats were used as the experimental group. The above-mentioned mussel mucin gel was continuously inhaled for 30 minutes. Another 10 samples were used as the control group, and no other treatment was performed before the test. After anesthesia, the rats are fixed on the rat plate and placed continuously next to the chemical forming steam. After 30 sec, a chemical irritative damage was formed.
  • the nasal cavity, mouth, pharynx, and throat mucosa of the inhalation injury were weighed and weighed, then placed in a 60 ° C oven for 48 hours, and the dry weight was weighed.
  • the mass of the mucosa after drying was divided by the mass of the mucosa before drying, and the ratio obtained was used to evaluate the degree of edema caused by the damage. The higher the ratio, the lighter the edema.
  • the dry weight/wet weight of the test sites of the 10 test rats was 37.2% ⁇ 2.2%; the dry weight/wet weight of the test sites of the 10 control rats was 19.3% ⁇ 2.6%.
  • the edema of the test site in which the mussel mucin gel drug was inhaled was significantly lighter than that of the control group. It is indicated that the mussel mucin product has the function of protecting the mucosa from chemical stimulation.
  • Example 14 Application of Mussel Mucin Foam Drugs in Chemical Stimulation Protection.
  • mice Twenty white rats weighing 180g-200g and 10 rats were used as the experimental group. The above-mentioned mussel mucin liquid medicine was inhaled continuously for 30 minutes. Another 10 samples were used as the control group, and no other treatment was performed before the test. After anesthesia, the rats were fixed on a rat plate and placed under the vapor of chemical reagents for 30 sec to form a chemically irritating lesion.
  • the nasal cavity, mouth, pharynx, and throat mucosa of the inhalation injury were weighed and weighed, then placed in a 60 ° C oven for 48 hours, and the dry weight was weighed.
  • the mass of the mucosa after drying was divided by the mass of the mucosa before drying, and the ratio obtained was used to evaluate the degree of edema caused by the damage. The higher the ratio, the lighter the edema.
  • the dry weight/wet weight of the test sites of the 10 test rats was 39.3% ⁇ 2.4%; the dry weight/wet weight of the test sites of the 10 control rats was 19.3% ⁇ 2.6%.
  • the edema of the test site in which the mussel mucin foam drug was inhaled was significantly lighter than that of the control group. It is indicated that the mussel mucin product has the function of protecting the mucosa from chemical stimulation.
  • Example 15 Application of mussel mucin gel disinfectant product in environmental pollution protection (PM2.5).
  • hydroxypropyl methylcellulose 10 g was taken, and 20 ml of deionized water was added thereto, and the mixture was heated at 90 ° C for 30 minutes to completely dissolve to obtain a gel matrix.
  • 40 ml of a mussel mucin solution having a concentration of 10.0 mg/ml was prepared to prepare a mussel mucin gel disinfecting product in which the mussel mucin concentration was 2.0 mg/ml.
  • Example 16 Application of mussel mucin foam disinfectant product in environmental pollution protection (PM2.5).
  • hydroxypropyl methylcellulose 10 g was taken, 20 ml of deionized water, 0.5 ml of Tween 60, and a bath at 90 ° C for 30 min to completely dissolve to obtain a foaming agent base.
  • 40 ml of a mussel mucin solution having a concentration of 10.0 mg/ml was prepared to prepare a mussel mucin foam disinfecting product in which the mussel mucin concentration was 2.0 mg/ml.
  • Example 17 Application of mussel mucin liquid medical device in ultraviolet protection.
  • a daylight simulator xenon arc lamp was used as a light source to continuously generate ultraviolet rays having a wavelength of 290 nm to 400 nm, and all the rats were placed under ultraviolet light irradiation once a day for 2 hours.
  • Example 18 Application of mussel mucin foam medical device in ultraviolet protection.
  • a daylight simulator xenon arc lamp was used as a light source to continuously generate ultraviolet rays having a wavelength of 290 nm to 400 nm, and all the rats were placed under ultraviolet light irradiation once a day for 2 hours.
  • Example 19 Application of mussel mucin gel cosmetics in radioactive ray protection.
  • Twenty Wistar rats weighing 200 g-300 g and 10 were used as test groups, and the above-mentioned mussel mucin gel cosmetics were dropped to the conjunctiva every day; 10 were used as the control group, and no other treatment was performed before the test.
  • the rats were irradiated with radioactive rays at a radiation intensity of 10 mV, and all the rats were placed under irradiation conditions for one time each time for 40 minutes.
  • Example 20 Application of mussel mucin foam cosmetic in radioactive ray protection.
  • Twenty Wistar rats weighing 200 g-300 g were used as the test group, and the above-mentioned mussel mucin foaming agent was applied to the conjunctiva every day. Ten other samples were used as the control group, and no other treatment was performed before the test. The rats were irradiated with radioactive rays at a radiation intensity of 10 mV, and all the rats were placed under irradiation conditions for one time each time for 40 minutes.
  • Example 21 Application of mussel mucin foam medical device in ultraviolet protection.
  • Example 22 Application of mussel mucin foaming agent in gastric mucosal protection.
  • test group took mussel mucin foam drugs half an hour before taking the drug, and the control group took the drugs normally.
  • the gastric mucosa of the subjects was tested.
  • the gastric mucosa of the 5 subjects in the test group was normal.
  • Two of the 5 subjects in the control group had symptoms of gastric mucosal congestion, 2 had edema, and 1 had ulceration. symptom.
  • mussel mucin foam medicine can be used for the protection of gastric mucosa.
  • Example 23 Application of mussel mucin foam disinfectant products in environmental pollution protection.
  • hydroxypropyl methylcellulose 10 g was taken, 20 ml of deionized water, 0.5 ml of Tween 60, and a bath at 90 ° C for 30 min to completely dissolve to obtain a foaming agent base.
  • 40 ml of a mussel mucin solution having a concentration of 10.0 mg/ml was prepared to prepare a mussel mucin foam disinfecting product in which the mussel mucin concentration was 2.0 mg/ml.
  • a foaming agent matrix to which no mussel mucin was added was prepared as a blank group.

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Abstract

本发明公开了贻贝粘蛋白或其制剂在防护粘膜的应用。具体地公开了贻贝粘蛋白或其制剂在防护粘膜免受紫外线损伤等的应用；防护粘膜免受电磁、射线、电脑辐射等放射线损伤等的应用；防护粘膜免受高温物体等的应用；防护粘膜免受低温物体等的应用；防护粘膜免受海水损伤等的应用；防护粘膜免受环境污染等损伤的应用；防护粘膜免受化学试剂刺激及刺激性药品、食品损伤的应用。

Description

贻贝粘蛋白用于粘膜的防护
本申请要求2015年8月14日递交的PCT/CN2015/087013的优先权，在其援引其全部内容并入本文。
技术领域
本发明大体涉及日化产品、化妆品、医疗器械、药品、消毒产品、保健品等领域，并且涉及一种贻贝粘蛋白或其制剂在防护粘膜中的应用。
背景技术
粘膜是体内的消化、呼吸、排泄、生殖等各器官的内壁，由粘液保持其表面湿润，具体地包括口腔、鼻腔、肠管、阴道、肠道等与外界相通体腔的湿润衬里。
皮肤和粘膜保护人体，使人体对外界形成密闭的系统，当有害物质侵入人体时，首先是皮肤和粘膜将外界致病因素阻挡在体外。所以皮肤和粘膜是人体抗感染的第一道防线。
粘膜易于受到物理因素、化学因素、细菌或其毒素的刺激。物理因素包括紫外线、放射线(如电磁、射线、电脑辐射等)、过冷、过烫、海水、环境污染等。化学因素包括化学试剂刺激及刺激性药品、食品等。
日光中的中波紫外线(UVB)在强烈照射时会引起粘膜的红、肿、灼热、灼痛、水肿等损伤。
包括放射性治疗在内的电离辐射等射线对恶性肿瘤有一定治疗作用，但在治疗期间会出现一定程度的粘膜反应，例如：红斑、烧灼和刺痒感、充血、水肿、渗出、糜烂、溃疡、毛细血管扩张、淋巴引流障碍等。
烫伤是由于吸入高热蒸气、误饮沸水、头颈烧伤者因吸入火焰或干热空气而造成的粘膜红肿、水疱组织坏死、溃烂、溃疡等问题。
冻伤是由于寒冷潮湿作用引起的粘膜的局部损伤，表现为冰凉、苍白、坚硬、感觉麻木或丧失、红肿充血、热、痒、灼痛、水疱、水肿、剧痛和组织坏死等。
海水中的盐分和游泳池中的氯气都会损伤皮肤粘膜，进而引发口腔粘膜 炎、鼻炎、咽喉炎等炎症反应，也会令头发变得干枯。
空气中的主要污染物有二氧化硫、氮氧化物、粒子状污染物、酸雨。二氧化硫(SO2)、一氧化二氮(N2O)、一氧化氮(NO)、二氧化氮(NO2)、三氧化二氮(N2O3)等含氮污染物对人体的粘膜有强烈刺激性。颗粒状污染物数量大且成分复杂，它本身可以是有毒物质或是其它污染物的运载体，易引发粘膜炎症等问题。
吸烟、喝酒、浓茶、咖啡、刺激性药品、食品、化学试剂等会对消化道、呼吸道粘膜造成损伤。
贻贝粘蛋白(Mussel adhesive protein，MAP)，也称作贻贝足丝蛋白(Mytilus edulis foot protein，Mefp)，是海洋贝类紫贻贝(Mytilus edulis Linnaeus)、厚壳贻贝(Mytilus coruscus)、翡翠贻贝(Perna viridis)等分泌的一种特殊的蛋白质。贻贝通常成群地附着在海岸边的礁石上或者轮船的底部，有在近海耐受波浪冲击的能力。实际上贻贝几乎可以极其牢固地附着在任何材料的基底上，如金属、木材、玻璃等。贻贝具有上述特性的主要原因是其足丝腺内可生成并储存这种特殊的粘蛋白，贻贝通过足丝释放粘蛋白到岩石一类的固体表面上，形成抗水的结合，从而将自己固定。
目前从贻贝中鉴定得到11种粘蛋白亚类，包括mefp1、mefp-2、mefp-3、mefp-4、mefp-5、mefp-6、胶原蛋白pre-COL-P、pre-COL-D、pre-COL-NG、足丝基质蛋白PTMP和DTMP(朱曜曜等，海洋科学进展，2014，32(4)：560-568)。贻贝粘蛋白具有2个结构特点：(1)含有赖氨酸，使蛋白带有高载量正电荷；(2)含3，4二羟基苯丙氨酸(DOPA，多巴)。人体的细胞和组织带有负电荷。贻贝粘蛋白通过自身正电荷与人体的细胞和组织负电荷之间的静电相互作用与细胞和组织紧密结合，发挥防护和治疗的作用。此外，多巴氧化生成邻位二醌，可以和未被氧化的多巴相互交联形成膜或是网状支架，促使蛋白质更加紧密、稳固地附着在人体表面，起到保护作用。贻贝粘蛋白是大分子蛋白质，在人体内完全降解的时间约为3-10天，其附着于细胞组织的能力优异，使贻贝粘蛋白可以稳固于局部，持续发挥作用。
虽然贻贝粘蛋白具有以上特点，但目前其产品应用领域非常有限。商品化的贻贝粘蛋白产品有美国BD Biosciences公司的Cell-Tak，韩国Kollodis的MAP Trix和瑞典Biopolymer的Hydrogel。这些产品或者是以贻贝粘蛋白 溶液状态直接使用，或者是以冻干粉制剂保存而在使用前溶解，它们的主要应用领域限于微观的细胞粘附和组织粘合剂。也有报道贻贝粘蛋白用于胎膜修复、作为抗海水腐蚀涂层、心脏载药支架等应用。
发明内容
本发明的目的是提供一种贻贝粘蛋白产品。
在本文中使用的贻贝粘蛋白是指从贻贝科(Mytilidae)双壳类软体动物中的紫贻贝(Mytilus edulis Linnaeus)、厚壳贻贝(Mytilus coruscus)或翡翠贻贝(Perna viridis)等海洋贻贝中纯化获得的、目前已知的贻贝粘蛋白11个亚类：mefp1、mefp-2、mefp-3、mefp-4、mefp-5、mefp-6、胶原蛋白pre-COL-P、pre-COL-D、pre-COL-NG、足丝基质蛋白PTMP和DTMP中的一种或几种的混合物。在本文中使用的贻贝粘蛋白在水溶液中的酸碱度可以是pH 1.0-7.0，特别是可以在pH 3.0-6.5的范围内以使其治疗效果更佳。
在本文中使用的贻贝粘蛋白可以采用以下制备方法获得，例如中国专利号ZL200710179491.0的“一种使用混合吸附色谱分离纯化贻贝粘蛋白的方法”，中国专利号ZL200710179492.5的“一种使用羧甲基离子交换色谱纯化贻贝粘蛋白的方法”，中国专利号ZL200910087567.6的“一种使用盐析和透析分离纯化贻贝粘蛋白的方法”等。
在本文中使用的贻贝粘蛋白可以是溶液或冻干粉形式，特别是贻贝粘蛋白在产品中的浓度可以是0.1-15.0mg/ml，当浓度过低时，贻贝粘蛋白的功效不大，当浓度过高时，可因正电荷载量过高引起细胞毒性，从而不利于发挥防护作用。
在本文中使用的贻贝粘蛋白也可以与辅料结合而制备成液体剂。示例性的贻贝粘蛋白液体剂是将贻贝粘蛋白溶液母液或冻干粉溶解或稀释至一定浓度或pH值制得，溶解或稀释用溶液可以是水、生理盐水、磷酸盐溶液、醋酸盐溶液、硼酸盐溶液等等。最终产品中贻贝粘蛋白的酸碱度可以是pH 1.0-7.0，特别是可以在pH 3.0-6.5的范围内治疗效果更佳。
在本文中使用的贻贝粘蛋白也可以与辅料结合而制备成凝胶剂。示例性的贻贝粘蛋白凝胶剂是将贻贝粘蛋白溶液或冻干粉与凝胶基质材料混合制得，所述凝胶基质材料可以是选自纤维素衍生物、卡波姆和海藻酸盐、西黄 蓍胶、明胶、果胶、卡拉胶、结冷胶、淀粉、黄原胶、阳离子瓜尔胶、琼脂、非纤维素多糖、乙烯聚合物、丙烯酸树脂、聚乙烯醇或聚羧乙烯中之一或其任意组合。
在本文中使用的贻贝粘蛋白还可以制成泡沫剂。示例性的贻贝粘蛋白泡沫剂是由贻贝粘蛋白溶液或冻干粉与发泡剂基质混合制得，所述发泡剂基质材料可以包括羟丙甲基纤维素、明胶、聚乙二醇十二烷基硫酸钠、脂肪醇聚氧乙烯醚磺酸钠、玉米麸质粉、丙烯酰胺之一或其任意组合。特别有益的是，泡沫剂在消泡过程中释放压力，更有利于贻贝粘蛋白在患处的附着、渗透和起效。同时泡沫剂消泡时间长，延长了作用时间，使贻贝粘蛋白起效更加显著。
本领域技术人员可以根据不同病期的临床适应症和特点来选择使用上述剂型或其他合适的剂型。
以上所有制剂都可采用本领域周知的方法制备，详细的操作步骤可以参照例如《制剂学》。
在本文中使用的贻贝粘蛋白可以作为主要原料，采用药学上可接受的载体制备药品。所述药品可以是液体剂、凝胶剂、泡沫剂。所述药品可通过口服、含化(舌下)、灌注(直肠给药)、滴入(眼)、喷雾(口、鼻)、吸入(口、鼻)、喷涂(口、鼻、耳、宫颈等)、腔镜(宫腔、腹腔等)、定向局部缓释、靶向给药的方式使用，并且可以以低温或加热方式给予。
在本文中使用的贻贝粘蛋白可以作为主要原料，制备医疗器械。本文中使用的术语医疗器械是指直接或间接用于人体的材料及其他类似或相关的物品。所述医疗器械可以是液体剂、凝胶剂、泡沫剂。所述医疗器械可通过口服、含化(舌下)、灌注(直肠给药)、滴入(眼)、喷雾(口、鼻)、吸入(口、鼻)、喷涂(口、鼻、耳、宫颈等)、腔镜(宫腔、腹腔等)、定向局部缓释、靶向给药的方式使用，并且可以以低温或加热方式给予。
在本文中使用的贻贝粘蛋白可以作为主要原料，采用化妆品领域可接受的辅料制备化妆品。所述化妆品可以是液体剂、凝胶剂、泡沫剂。所述化妆品可通过口服、含化(舌下)、灌注(直肠给药)、滴入(眼)、喷雾(口、鼻)、吸入(口、鼻)、喷涂(口、鼻、耳、宫颈等)、腔镜(宫腔、腹腔等)、定向局部缓释、靶向给药的方式使用，并且可以以低温或加热方式给予。
在本文中使用的贻贝粘蛋白可以作为主要原料，采用消毒产品领域可接受的辅料制备消毒产品。本文中使用的术语消毒产品是指用化学、物理、生物的方法杀灭或者消除环境中病原微生物的消毒剂、消毒器械、卫生用品和一次性使用的医疗用品。所述消毒产品可以是液体剂、凝胶剂、泡沫剂。所述消毒产品可通过口服、含化(舌下)、灌注(直肠给药)、滴入(眼)、喷雾(口、鼻)、吸入(口、鼻)、喷涂(口、鼻、耳、宫颈等)、腔镜(宫腔、腹腔等)、定向局部缓释、靶向给药的方式使用，并且可以以低温或加热方式给予。
在本文中使用的贻贝粘蛋白可以作为主要原料，采用保健品或食品领域可接受的辅料制备保健品或食品。所述保健品或食品可以是液体剂、凝胶剂、泡沫剂。所述保健品或食品可通过口服、含化(舌下)、灌注(直肠给药)、滴入(眼)、喷雾(口、鼻)、吸入(口、鼻)、喷涂(口、鼻、耳、宫颈等)、腔镜(宫腔、腹腔等)、定向局部缓释、靶向给药的方式使用，并且可以以低温或加热方式给予。
在本文中使用的贻贝粘蛋白可以作为主要原料，采用日化产品领域可接受的辅料制备日化产品。本文中所使用的术语日化产品是指平日常用的科技化学制品，包括洗发水、沐浴露等。所述日化产品可以是液体剂、凝胶剂、泡沫剂。所述日化产品可通过口服、含化(舌下)、灌注(直肠给药)、滴入(眼)、喷雾(口、鼻)、吸入(口、鼻)、喷涂(口、鼻、耳、宫颈等)、腔镜(宫腔、腹腔等)、定向局部缓释、靶向给药的方式使用，并且可以以低温或加热方式给予。
本发明的另一个目的是提供贻贝粘蛋白产品在粘膜防护中的应用。
在本文中使用的术语“粘膜防护”是指通过防护措施使包括口腔、鼻腔、肠管、阴道、肠道等的粘膜免于受到物理和化学损伤。
特别是，本发明提供了贻贝粘蛋白或其制剂在防护粘膜免受紫外线损伤等中的应用。
特别是，本发明提供了贻贝粘蛋白或其制剂在防护粘膜免受放射线(电磁、射线、电脑辐射等)损伤等中的应用。
特别是，本发明提供了贻贝粘蛋白或其制剂在防护粘膜免受高温物体等引发的烫伤中的应用。
特别是，本发明提供了贻贝粘蛋白或其制剂在防护粘膜免受低温物体等引发的冻伤中的应用。
特别是，本发明提供了贻贝粘蛋白或其制剂在防护粘膜免受海水损伤中的应用；在防护粘膜免受环境污染等损伤中的应用。
特别是，本发明提供了贻贝粘蛋白或其制剂在防护粘膜免受吸烟、喝酒、浓茶、咖啡、刺激性药品、食品、化学试剂等损伤中的应用。
根据本发明的一个方面，本发明提供了一种用于防护粘膜免受吸烟、喝酒、浓茶、咖啡、刺激性药品、食品、化学试剂等损伤的含贻贝粘蛋白的液体剂。
根据本发明的又一个方面，本发明提供了一种用于防护粘膜免受吸烟、喝酒、浓茶、咖啡、刺激性药品、食品、化学试剂等损伤的含贻贝粘蛋白的凝胶剂。
根据本发明的又一个方面，本发明提供了一种用于防护粘膜免受吸烟、喝酒、浓茶、咖啡、刺激性药品、食品、化学试剂等损伤的含贻贝粘蛋白的泡沫剂。
根据本发明的一个方面，本发明提供了一种制备用于防护粘膜免受吸烟、喝酒、浓茶、咖啡、刺激性药品、食品、化学试剂等损伤的含贻贝粘蛋白的液体剂的方法。
根据本发明的一个方面，本发明提供了一种制备用于防护粘膜免受吸烟、喝酒、浓茶、咖啡、刺激性药品、食品、化学试剂等损伤的含贻贝粘蛋白的凝胶剂的方法。
根据本发明的一个方面，本发明提供了一种制备用于防护粘膜免受吸烟、喝酒、浓茶、咖啡、刺激性药品、食品、化学试剂等损伤的含贻贝粘蛋白的泡沫剂。其中，所述粘膜炎症可以包括如本文上面所述的任何一种。
具体实施方式
本发明的实施方式包括：
1、贻贝粘蛋白在防护粘膜损伤中的应用。
2、根据实施方式1的贻贝粘蛋白应用，其中所述贻贝粘蛋白可以是来自亚类：mefp1、mefp-2、mefp-3、mefp-4、mefp-5、mefp-6、胶原蛋白pre-COL-P、 pre-COL-D、pre-COL-NG、足丝基质蛋白PTMP和DTMP中的一种或几种的混合物。
3、根据实施方式1的贻贝粘蛋白应用，其中所述贻贝粘蛋白浓度可以是0.1-15.0mg/ml。
4、根据实施方式1的贻贝粘蛋白应用，其中所述贻贝粘蛋白可以是以液体剂、凝胶剂或泡沫剂使用。
5、根据实施方式1的贻贝粘蛋白应用，其中最终产品中的贻贝粘蛋白可以是在pH 1.0-7.0的范围内，特别是可以在pH 3.0-6.5的范围内。
6、根据实施方式1-4中任一项的贻贝粘蛋白应用，其中所述防护粘膜可以包括防护粘膜免受紫外线、放射线、高温物体、低温物体、海水、环境污染、化学试剂刺激及刺激性药品、食品的损伤。
7、贻贝粘蛋白作为活性成分在用于防护粘膜的组合物中的应用，其中所述组合物可以是以液体剂、凝胶剂或泡沫剂使用。
8、根据实施方式7的贻贝粘蛋白应用，其中所述组合物可以通过口服、含化(舌下)、灌注(直肠给药)、滴入(眼)、喷雾(口、鼻)、吸入(口、鼻)、喷涂(口、鼻、耳、宫颈等)、腔镜(宫腔、腹腔等)、定向局部缓释、靶向给药的方式使用。
9、根据实施方式7的贻贝粘蛋白应用，其中所述组合物可以以低温或加热方式给予。
10、贻贝粘蛋白作为活性成分在用于粘膜防护的药品中的应用。
11、贻贝粘蛋白作为活性成分在用于粘膜防护的化妆品中的应用。
12、贻贝粘蛋白作为活性成分在用于粘膜防护的消毒产品中的应用。
13、贻贝粘蛋白作为活性成分在用于粘膜防护的保健品或食品中的应用。
14、贻贝粘蛋白作为活性成分在用于粘膜防护的日化产品中的应用。
15、一种用于粘膜防护的凝胶剂，含有贻贝粘蛋白作为活性成分。
16、一种用于粘膜防护的液体剂，含有贻贝粘蛋白作为活性成分。
17、一种用于粘膜防护的泡沫剂，含有贻贝粘蛋白作为活性成分。特别地，所述防护粘膜可以包括防护粘膜免受紫外线、放射线、高温物体、低温物体、海水、环境污染、化学试剂刺激及刺激性药品、食品的损伤。
18、一种制备用于粘膜防护的凝胶剂的方法，可以包括：1)获取或制备 贻贝粘蛋白溶液或冻干粉，2)获取或制备凝胶基质，3)混合步骤1)的贻贝粘蛋白溶液和步骤2)的凝胶基质，以及4)调节pH至弱酸性，获得凝胶剂。
19、一种制备用于粘膜防护的液体剂的方法，可以包括：获取或制备贻贝粘蛋白溶液或冻干粉，用药学上可接受的稀释剂稀释，以及调节pH至弱酸性，获得液体剂。
20、一种制备用于粘膜防护的泡沫剂的方法，可以包括：)获取或制备贻贝粘蛋白溶液或冻干粉，与药学上可接受的发泡剂基质混合，及调节pH至弱酸性，获得泡沫剂。
下面将结合具体实施例对本发明作进一步说明。需要指出的是，由本发明中的贻贝粘蛋白或贻贝粘蛋白的各种制剂形成的药品、医疗器械、化妆品、消毒产品、保健品或食品、日化产品在施用于受试者后，都可以应用于上文所述的适应症并展现出上文所述的功能，在本发明范围内的所有剂型均已测试，下文中，仅仅是为说明，只在实施例中描述了其中一少部分，然而不应将其理解为对本发明的限制。
除非特殊说明，否则本发明中所使用的试剂都是市售可购买的。
实施例1：贻贝粘蛋白水凝胶药品在胃粘膜防护中的应用。
取明胶10g，加入100ml去离子水，90℃温浴30min至基质完全溶解，另取浓度为10.0mg/ml的贻贝粘蛋白溶液60ml，边搅拌边加入到凝胶基质中，混合均匀后形成贻贝粘蛋白水凝胶药品，其中贻贝粘蛋白浓度为10mg/g。
取健康雄性大鼠进行试验，每只180-200g，共20只，对照组和试验组各10只。对照组采用无水乙醇灌胃，每只1次，每次1.0ml。1小时后处死动物，暴露完整胃，结扎幽门，灌注10％甲醛溶液固定20分钟，然后沿胃大弯剪开，洗净胃内容物，展开胃粘膜，在体视解剖显微镜下用游标卡尺测量出血点或出血带的长度和宽度。试验组先服用上述贻贝粘蛋白水凝胶药品2.0ml，1小时后按照对照组相同的方法用无水乙醇灌胃，并测量出血点。对照组和试验组以胃粘膜损伤程度(评价方法参见表1)及损伤发生率评价胃粘膜损伤程度，从胃粘膜损伤程度和损伤发生率(参见表2)可以看出，使 用贻贝粘蛋白水凝胶药品可以降低胃粘膜损伤的发生率并降低损伤程度。
表1：急性酒精损伤观察评分标准
Figure PCTCN2016095362-appb-000001
损伤发生率(％)＝出现出血或溃疡的大鼠数量/大鼠总量×100％
表2：胃粘膜损伤程度及损伤发生率
分组 损伤发生率(％) 损伤程度
对照组 100 7.4±0.8
试验组 90 4.1±0.7
实施例2：贻贝粘蛋白液体保健品在胃粘膜防护中的应用。
取贻贝粘蛋白溶液，用0.5％乙酸配制成浓度为5.0mg/ml的水溶液，获得贻贝粘蛋白液体保健品。
取健康雄性大鼠进行试验，每只180-200g，共20只，对照组和试验组各10只。对照组采用无水乙醇灌胃，每只1次，每次1.0ml。1小时后处死动物，暴露完整胃，结扎幽门，灌注10％甲醛溶液固定20分钟，然后沿胃大弯剪开，洗净胃内容物，展开胃粘膜，在体视解剖显微镜下用游标卡尺测量出血点或出血带的长度和宽度。试验组先服用贻贝粘蛋白液体保健品3.0ml，1小时后按照对照组相同的方法用无水乙醇灌胃，并测量出血点。对照组和试验组以胃粘膜损伤程度(评价方法参见表3)及损伤发生率评价胃粘膜损伤程度，从胃粘膜损伤程度和损伤发生率(参见表4)可以看出，使用贻贝粘蛋白水凝胶药品可以降低胃粘膜损伤的发生率并降低损伤程度。
表3：急性酒精损伤观察评分标准
Figure PCTCN2016095362-appb-000002
Figure PCTCN2016095362-appb-000003
损伤发生率(％)＝出现出血或溃疡的大鼠数量/大鼠总量×100％
表4：胃粘膜损伤程度及损伤发生率
分组 损伤发生率(％) 损伤程度
对照组 100 7.6±0.7
试验组 90 5.3±0.4
实施例3：贻贝粘蛋白水凝胶医疗器械在高温吸入性损伤防护中的应用。
取浓度为5.0mg/ml的贻贝粘蛋白溶液50ml，用生理盐水稀释，制备成贻贝粘蛋白液体医疗器械，其中贻贝粘蛋白浓度为2.5mg/ml。
取20只体重在180g-200g的白色大鼠，10只作为试验组，连续吸入雾化的上述贻贝粘蛋白液体医疗器械30min；另取10只作为对照组，试验前不作其它处理。大鼠麻醉后，固定于鼠板上，在高温蒸汽旁边持续放置30sec，形成吸入性损伤。
24h后剪下吸入性损伤鼻腔、口腔、咽、喉部粘膜秤量湿重，然后放入60℃烤箱烤干48h，称量干重。将烘干之后的粘膜质量除以烘干之前的粘膜质量，得到的比值用于评估损伤造成的水肿程度，比值越高表明水肿越轻。
10只试验组大鼠受试部位的干重/湿重，平均为23.4％±1.8％；10只对照组大鼠受试部位的干重/湿重，平均为12.9％±1.2％。
涂布了贻贝粘蛋白液体医疗器械的受试部位水肿情况明显比对照组轻。表明贻贝粘蛋白产品具有防护粘膜免受高温气体侵害的作用。
实施例4：贻贝粘蛋白液体药品在化学刺激防护中的应用。
取浓度为10.0mg/ml的贻贝粘蛋白溶液40ml，用生理盐水稀释，制备成贻贝粘蛋白液体药品，其中贻贝粘蛋白浓度为5.0mg/ml。
取20只体重在180g-200g的白色大鼠，10只作为试验组，连续吸入雾化的上述贻贝粘蛋白液体药品30min；另取10只作为对照组，试验前不作其它处理。大鼠麻醉后，固定于鼠板上，在化学试剂形成蒸汽旁边持续放置30sec，形成化学刺激性损伤。
24h后剪下吸入性损伤鼻腔、口腔、咽、喉部粘膜秤量湿重，然后放入 60℃烤箱烤干48h，称量干重。将烘干之后的粘膜质量除以烘干之前的粘膜质量，得到的比值用于评估损伤造成的水肿程度，比值越高表明水肿越轻。
10只试验组大鼠受试部位的干重/湿重，平均为35.1％±2.9％；10只对照组大鼠受试部位的干重/湿重，平均为19.3％±2.6％。
吸入了贻贝粘蛋白液体药品的受试部位水肿情况明显比对照组轻。表明贻贝粘蛋白产品具有防护粘膜免受化学刺激侵害的作用。
实施例5：贻贝粘蛋白液体消毒产品在环境污染防护(PM2.5)中的应用。
取浓度为10.0mg/ml的贻贝粘蛋白溶液40ml，用生理盐水稀释，制备成贻贝粘蛋白液体消毒产品，其中贻贝粘蛋白浓度为2.0mg/ml。
取20只体重在200g-300g的Wistar大鼠，取10只作为试验组，每天用上述贻贝粘蛋白消毒产品清洗鼻腔；另取10只作为对照组，试验前不作其它处理。将全部大鼠放置在PM 2.5含量超过300的环境中饲养。分别在饲养10、20、30天观察大鼠因环境污染引起的变应性鼻炎的症状(参见表5)，发现采用贻贝粘蛋白清洁鼻腔的大鼠打喷嚏和搔鼻次数明显低于对照组，表明贻贝粘蛋白液体消毒产品具有防护环境污染侵害的作用。
表5：受试大鼠打喷嚏和搔鼻次数
Figure PCTCN2016095362-appb-000004
实施例6：贻贝粘蛋白水凝胶医疗器械在紫外线防护中的应用。
取明胶10g，加入100ml去离子水，90℃温浴30min至基质完全溶解，另取浓度为20.0mg/ml的贻贝粘蛋白溶液60ml，边搅拌边加入到凝胶基质中，用生理盐水调整浓度，混合均匀后形成贻贝粘蛋白水凝胶医疗器械，其中贻贝粘蛋白浓度为10mg/g。
取20只体重在200g-300g的Wistar大鼠，10只作为试验组，每天用上述贻贝粘蛋白水凝胶医疗器械涂抹结膜；另取10只作为对照组，试验前不 作其它处理。采用日光模拟器氙弧灯作为光源，连续产生波长为290nm-400nm的紫外线，将全部大鼠放置在紫外光照射条件下，每日1次，每次2小时。
对照组大鼠从第2天开始有4只大鼠出现结膜充血、分泌物增多现象，占全部测试数量的40％；至第3天全部大鼠均出现结膜充血、分泌物增多现象。试验组大鼠在测试第2天有0只出现结膜充血、分泌物增多现象，至第3天有3只出现结膜充血、分泌物增多现象，占总数的30％。表明贻贝粘蛋白水凝胶医疗器械具有防护粘膜免受紫外线侵害的作用。
实施例7：贻贝粘蛋白液体化妆品在放射性射线防护中的应用。
取浓度为10.0mg/ml的贻贝粘蛋白溶液50ml，用生理盐水稀释，制备成贻贝粘蛋白液体化妆品，其中贻贝粘蛋白浓度为2mg/ml。
取20只体重在200g-300g的Wistar大鼠，10只作为试验组，每天用上述贻贝粘蛋白液体化妆品滴至结膜；另取10只作为对照组，试验前不作其它处理。采用放射性射线照射大鼠，射线强度为10mV，将全部大鼠放置在射线照射条件下，每日1次，每次40分钟。
对照组大鼠从第2天开始有5只大鼠出现结膜充血、分泌物增多现象，占全部测试数量的50％；至第3天全部大鼠均出现结膜充血、分泌物增多现象。试验组大鼠在测试第2天有1只出现结膜充血、分泌物增多现象，至第3天有5只出现结膜充血、分泌物增多现象，占总数的50％。表明贻贝粘蛋白液体化妆品具有防护粘膜免受放射性射线侵害的作用。
实施例8：贻贝粘蛋白泡沫剂医疗器械在阴道及宫颈粘膜防护中的应用
取贻贝粘蛋白溶液，与羟丙甲基纤维素按照质量比2∶1的比例混合，用柠檬酸调整酸碱度为pH 5.0，获得贻贝粘蛋白泡沫剂医疗器械，其中贻贝粘蛋白浓度为5mg/mL。
取体重为200-250g雌性大鼠20只，按体重均分为试验组和对照组。对照组采用苯酚胶浆(苯酚5ml，西黄蓍胶1g，甘油4ml，蒸馏水20ml)于每鼠阴道内按0.01mg/kg体重灌注，每日1次，使用2d。试验组采用上述贻贝粘蛋白泡沫剂医疗器械喷涂于阴道及宫颈区域，用量为1ml/鼠。1小 时后，采用与对照组相同的处理方法灌注苯酚胶浆。按上述分组分别给药，第10天称重后处死，取阴道及宫颈称重，并计算脏器指数(表6)。试验组的脏器指数低于对照组，证明试验组大鼠阴道及粘膜炎症反应程度低于对照组。可见本发明的贻贝粘蛋白产品具有防护阴道及宫颈粘膜的作用。
表6：受试大鼠阴道和宫颈脏器指数
分组 阴道和宫颈脏器指数(g/kg)
对照组 0.055±0.003
试验组 0.032±0.004
实施例9：贻贝粘蛋白凝胶医疗器械在阴道及宫颈粘膜防护中的应用
取贻贝粘蛋白溶液，与海藻酸钠按照质量比2∶1的比例混合，用柠檬酸调整酸碱度为pH 5.0，获得贻贝粘蛋白凝胶医疗器械，其中贻贝粘蛋白浓度为5mg/mL。
取体重为200-250g雌性大鼠20只，按体重均分为试验组和对照组。对照组采用苯酚胶浆(苯酚5ml，西黄蓍胶1g，甘油4ml，蒸馏水20ml)于每鼠阴道内按0.01mg/kg体重灌注，每日1次，使用2d。试验组采用上述贻贝粘蛋白凝胶医疗器械喷涂于阴道及宫颈区域，用量为1ml/鼠。1小时后，采用与对照组相同的处理方法灌注苯酚胶浆。按上述分组分别给药，第10天称重后处死，取阴道及宫颈称重，并计算脏器指数(表7)。试验组的脏器指数低于对照组，证明试验组大鼠阴道及粘膜炎症反应程度低于对照组。可见本发明的贻贝粘蛋白产品具有防护阴道及宫颈粘膜的作用。
表7：受试大鼠阴道和宫颈脏器指数
分组 阴道和宫颈脏器指数(g/kg)
对照组 0.053±0.003
试验组 0.041±0.004
实施例10：贻贝粘蛋白泡沫剂药品在胃粘膜防护中的应用。
取明胶10g，吐温0.5ml，加入100ml去离子水，90℃温浴30min至基质完全溶解，另取浓度为10.0mg/ml的贻贝粘蛋白溶液60ml，边搅拌边加入到泡沫基质中，混合均匀后形成贻贝粘蛋白泡沫剂药品，其中贻贝粘蛋白 浓度为10mg/g。
取健康雄性大鼠进行试验，每只180-200g，共20只，对照组和试验组各10只。对照组采用无水乙醇灌胃，每只1次，每次1.0ml。1小时后处死动物，暴露完整胃，结扎幽门，灌注10％甲醛溶液固定20分钟，然后沿胃大弯剪开，洗净胃内容物，展开胃粘膜，在体视解剖显微镜下用游标卡尺测量出血点或出血带的长度和宽度。试验组先服用上述贻贝粘蛋白水凝胶药品2.0ml，1小时后按照与对照组相同的方法用无水乙醇灌胃，并测量出血点。对照组和试验组以胃粘膜损伤程度(评价方法参见表1)及损伤发生率评价胃粘膜损伤程度，从胃粘膜损伤程度和损伤发生率(参见表8)可以看出，使用贻贝粘蛋白泡沫剂药品可以降低胃粘膜损伤的发生率并降低损伤程度。
表1：急性酒精损伤观察评分标准
Figure PCTCN2016095362-appb-000005
损伤发生率(％)＝出现出血或溃疡的大鼠数量/大鼠总量×100％
表8：胃粘膜损伤程度及损伤发生率
分组 损伤发生率(％) 损伤程度
对照组 100 7.4±0.8
试验组 83 3.6±0.6
实施例11：贻贝粘蛋白液体医疗器械在高温吸入性损伤防护中的应用。
取浓度为5.0mg/ml的贻贝粘蛋白溶液50ml，用生理盐水稀释，制备成贻贝粘蛋白液体医疗器械，其中贻贝粘蛋白浓度为2.5mg/ml。
取20只体重在180g-200g的白色大鼠，10只作为试验组，连续吸入雾化的上述贻贝粘蛋白液体医疗器械30min；另取10只作为对照组，试验前不作其它处理。大鼠麻醉后，固定于鼠板上，在高温蒸汽旁边持续放置30sec，形成吸入性损伤。
24h后剪下吸入性损伤鼻腔、口腔、咽、喉部粘膜秤量湿重，然后放入 60℃烤箱烤干48h，称量干重。将烘干之后的粘膜质量除以烘干之前的粘膜质量，得到的比值用于评估损伤造成的水肿程度，比值越高表明水肿越轻。
10只试验组大鼠受试部位的干重/湿重，平均为16.7％±1.3％；10只对照组大鼠受试部位的干重/湿重，平均为12.9％±1.2％。
涂布了贻贝粘蛋白液体医疗器械的受试部位水肿情况明显比对照组轻。表明贻贝粘蛋白产品具有防护粘膜免受高温气体侵害的作用。
实施例12：贻贝粘蛋白泡沫剂医疗器械在高温吸入性损伤防护中的应用。
取浓度为5.0mg/ml的贻贝粘蛋白溶液50ml，加入到等体积的甲基纤维素泡沫剂基质中，制备成贻贝粘蛋白泡沫剂医疗器械，其中贻贝粘蛋白浓度为2.5mg/ml。
取20只体重在180g-200g的白色大鼠，10只作为试验组，连续吸入雾化的上述贻贝粘蛋白液体医疗器械30min；另取10只作为对照组，试验前不作其它处理。大鼠麻醉后，固定于鼠板上，在高温蒸汽旁边持续放置30sec，形成吸入性损伤。
24h后剪下吸入性损伤鼻腔、口腔、咽、喉部粘膜秤量湿重，然后放入60℃烤箱烤干48h，称量干重。将烘干之后的粘膜质量除以烘干之前的粘膜质量，得到的比值用于评估损伤造成的水肿程度，比值越高表明水肿越轻。
10只试验组大鼠受试部位的干重/湿重，平均为29.1％±2.0％；10只对照组大鼠受试部位的干重/湿重，平均为12.9％±1.2％。
涂布了贻贝粘蛋白液体医疗器械的受试部位水肿情况明显比对照组轻。表明贻贝粘蛋白产品具有防护粘膜免受高温气体侵害的作用。
实施例13：贻贝粘蛋白凝胶剂药品在化学刺激防护中的应用。
取羟丙基甲基纤维素10g，加入20ml去离子水，90℃温浴30min至完全溶解得到凝胶剂基质。另取浓度为10.0mg/ml的贻贝粘蛋白溶液40ml，制备成贻贝粘蛋白凝胶剂药品，其中贻贝粘蛋白浓度为5.0mg/ml。
取20只体重在180g-200g的白色大鼠，10只作为试验组，连续吸入雾化的上述贻贝粘蛋白凝胶齐药品30min；另取10只作为对照组，试验前不作其它处理。大鼠麻醉后，固定于鼠板上，在化学试剂形成蒸汽旁边持续放 置30sec，形成化学刺激性损伤。
24h后剪下吸入性损伤鼻腔、口腔、咽、喉部粘膜秤量湿重，然后放入60℃烤箱烤干48h，称量干重。将烘干之后的粘膜质量除以烘干之前的粘膜质量，得到的比值用于评估损伤造成的水肿程度，比值越高表明水肿越轻。
10只试验组大鼠受试部位的干重/湿重，平均为37.2％±2.2％；10只对照组大鼠受试部位的干重/湿重，平均为19.3％±2.6％。
吸入了贻贝粘蛋白凝胶药品的受试部位水肿情况明显比对照组轻。表明贻贝粘蛋白产品具有防护粘膜免受化学刺激侵害的作用。
实施例14：贻贝粘蛋白泡沫剂药品在化学刺激防护中的应用。
取羟丙基甲基纤维素10g，加入20ml去离子水，0.5ml吐温60，90℃温浴30min至完全溶解得到泡沫剂基质。另取浓度为10.0mg/ml的贻贝粘蛋白溶液40ml，制备成贻贝粘蛋白泡沫剂药品，其中贻贝粘蛋白浓度为5.0mg/ml。
取20只体重在180g-200g的白色大鼠，10只作为试验组，连续吸入雾化的上述贻贝粘蛋白液体药品30min；另取10只作为对照组，试验前不作其它处理。大鼠麻醉后，固定于鼠板上，在化学试剂形成蒸汽旁边持续放置30sec，形成化学刺激性损伤。
24h后剪下吸入性损伤鼻腔、口腔、咽、喉部粘膜秤量湿重，然后放入60℃烤箱烤干48h，称量干重。将烘干之后的粘膜质量除以烘干之前的粘膜质量，得到的比值用于评估损伤造成的水肿程度，比值越高表明水肿越轻。
10只试验组大鼠受试部位的干重/湿重，平均为39.3％±2.4％；10只对照组大鼠受试部位的干重/湿重，平均为19.3％±2.6％。
吸入了贻贝粘蛋白泡沫剂药品的受试部位水肿情况明显比对照组轻。表明贻贝粘蛋白产品具有防护粘膜免受化学刺激侵害的作用。
实施例15：贻贝粘蛋白凝胶剂消毒产品在环境污染防护(PM2.5)中的应用。
取羟丙基甲基纤维素10g，加入20ml去离子水，90℃温浴30min至完全溶解得到凝胶基质。取浓度为10.0mg/ml的贻贝粘蛋白溶液40ml，制备成贻贝粘蛋白凝胶剂消毒产品，其中贻贝粘蛋白浓度为2.0mg/ml。
取20只体重在200g-300g的Wistar大鼠，取10只作为试验组，每天用上述贻贝粘蛋白消毒产品喷涂鼻腔；另取10只作为对照组，试验前不作其它处理。将全部大鼠放置在PM 2.5含量超过300的环境中饲养。分别在饲养10、20、30天观察大鼠因环境污染引起的变应性鼻炎的症状(参见表9)，发现采用贻贝粘蛋白喷涂鼻腔的大鼠打喷嚏和搔鼻次数明显低于对照组，表明贻贝粘蛋白凝胶消毒产品具有防护环境污染侵害的作用。
表9：受试大鼠打喷嚏和搔鼻次数
Figure PCTCN2016095362-appb-000006
实施例16：贻贝粘蛋白泡沫剂消毒产品在环境污染防护(PM2.5)中的应用。
取羟丙基甲基纤维素10g，加入20ml去离子水，0.5ml吐温60，90℃温浴30min至完全溶解得到泡沫剂基质。取浓度为10.0mg/ml的贻贝粘蛋白溶液40ml，制备成贻贝粘蛋白泡沫消毒产品，其中贻贝粘蛋白浓度为2.0mg/ml。
取20只体重在200g-300g的Wistar大鼠，取10只作为试验组，每天用上述贻贝粘蛋白消毒产品喷涂鼻腔；另取10只作为对照组，试验前不作其它处理。将全部大鼠放置在PM 2.5含量超过300的环境中饲养。分别在饲养10、20、30天观察大鼠因环境污染引起的变应性鼻炎的症状(参见表10)，发现采用贻贝粘蛋白喷涂鼻腔的大鼠打喷嚏和搔鼻次数明显低于对照组，表明贻贝粘蛋白泡沫剂消毒产品具有防护环境污染侵害的作用。
表10：受试大鼠打喷嚏和搔鼻次数
Figure PCTCN2016095362-appb-000007
实施例17：贻贝粘蛋白液体医疗器械在紫外线防护中的应用。
取浓度为20.0mg/ml的贻贝粘蛋白溶液60ml，用生理盐水调整浓度，混合均匀后形成贻贝粘蛋白液体医疗器械，其中贻贝粘蛋白浓度为10mg/g。
取20只体重在200g-300g的Wistar大鼠，10只作为试验组，每天用上述贻贝粘蛋白液体医疗器械喷涂结膜；另取10只作为对照组，试验前不作其它处理。采用日光模拟器氙弧灯作为光源，连续产生波长为290nm-400nm的紫外线，将全部大鼠放置在紫外光照射条件下，每日1次，每次2小时。
对照组大鼠从第2天开始有4只大鼠出现结膜充血、分泌物增多现象，占全部测试数量的40％；至第3天全部大鼠均出现结膜充血、分泌物增多现象。试验组大鼠在测试第2天有0只出现结膜充血、分泌物增多现象，至第3天有5只出现结膜充血、分泌物增多现象，占总数的50％。表明贻贝粘蛋白液体医疗器械具有防护粘膜免受紫外线侵害的作用。
实施例18：贻贝粘蛋白泡沫剂医疗器械在紫外线防护中的应用。
取明胶10g，丙三醇2ml，加入100ml去离子水，90℃温浴30min至基质完全溶解，取浓度为20.0mg/ml的贻贝粘蛋白溶液60ml，混合均匀后形成贻贝粘蛋白泡沫剂医疗器械，其中贻贝粘蛋白浓度为10mg/g。
取20只体重在200g-300g的Wistar大鼠，10只作为试验组，每天用上述贻贝粘蛋白泡沫剂医疗器械涂抹结膜；另取10只作为对照组，试验前不作其它处理。采用日光模拟器氙弧灯作为光源，连续产生波长为290nm-400nm的紫外线，将全部大鼠放置在紫外光照射条件下，每日1次，每次2小时。
对照组大鼠从第2天开始有4只大鼠出现结膜充血、分泌物增多现象，占全部测试数量的40％；至第3天全部大鼠均出现结膜充血、分泌物增多现象。试验组大鼠在测试第2天有0只出现结膜充血、分泌物增多现象，至第3天有2只出现结膜充血、分泌物增多现象，占总数的20％。表明贻贝粘蛋白泡沫剂医疗器械具有防护粘膜免受紫外线侵害的作用。
实施例19：贻贝粘蛋白凝胶化妆品在放射性射线防护中的应用。
取贻贝粘蛋白溶液，与瓜尔胶、丙三醇按体积比2∶1∶1混合，加入注射 水，制备成贻贝粘蛋白凝胶化妆品，其中贻贝粘蛋白浓度为2mg/ml。
取20只体重在200g-300g的Wistar大鼠，10只作为试验组，每天用上述贻贝粘蛋白凝胶化妆品滴至结膜；另取10只作为对照组，试验前不作其它处理。采用放射性射线照射大鼠，射线强度为10mV，将全部大鼠放置在射线照射条件下，每日1次，每次40分钟。
对照组大鼠从第2天开始有5只大鼠出现结膜充血、分泌物增多现象，占全部测试数量的50％；至第3天全部大鼠均出现结膜充血、分泌物增多现象。试验组大鼠在测试第2天有1只出现结膜充血、分泌物增多现象，至第3天有4只出现结膜充血、分泌物增多现象，占总数的40％。表明贻贝粘蛋白凝胶化妆品具有防护粘膜免受放射性射线侵害的作用。
实施例20：贻贝粘蛋白泡沫剂化妆品在放射性射线防护中的应用。
取贻贝粘蛋白溶液，与瓜尔胶、丙三醇、吐温60按体积比2∶1∶1∶0.5混合，加入注射水，制备成贻贝粘蛋白泡沫剂化妆品，其中贻贝粘蛋白浓度为2mg/ml。
取20只体重在200g-300g的Wistar大鼠，10只作为试验组，每天用上述贻贝粘蛋白泡沫剂化妆品滴至结膜；另取10只作为对照组，试验前不作其它处理。采用放射性射线照射大鼠，射线强度为10mV，将全部大鼠放置在射线照射条件下，每日1次，每次40分钟。
对照组大鼠从第2天开始有5只大鼠出现结膜充血、分泌物增多现象，占全部测试数量的50％；至第3天全部大鼠均出现结膜充血、分泌物增多现象。试验组大鼠在测试第2天有0只出现结膜充血、分泌物增多现象，至第3天有3只出现结膜充血、分泌物增多现象，占总数的30％。表明贻贝粘蛋白泡沫剂化妆品具有防护粘膜免受放射性射线侵害的作用。
实施例21：贻贝粘蛋白泡沫剂医疗器械在紫外线防护中的应用。
取明胶10g，丙三醇2ml，加入100ml去离子水，90℃温浴30min至基质完全溶解，取浓度为20.0mg/ml的贻贝粘蛋白溶液60ml，混合均匀后形成贻贝粘蛋白泡沫剂医疗器械，其中贻贝粘蛋白浓度为2mg/g。另取不加入贻贝粘蛋白的泡沫剂基质作为对照。
收集20例既往电光性眼炎患者，年龄20-40岁之间，职业特点：每日在强紫外线下工作时间不少于6小时。20名志愿者一只眼睛使用试验组贻贝粘蛋白泡沫剂医疗器械，一只眼睛使用对照组产品，10天后对受试者进行眼部检测。使用10天后试验组均未出现烧灼感，畏光、流泪、眼脸痉挛，头痛，眼睑及面部皮肤潮红和灼痛感，眼裂部结膜充血、水肿等症状。对照组有11名志愿者出现眼部结膜充血、水肿现象，3名志愿者出现畏光、流泪症状，2名志愿者出现灼痛感症状。试验结果表明贻贝粘蛋白泡沫剂医疗器械具有防护粘膜免受紫外线侵害的作用。
实施例22：贻贝粘蛋白泡沫剂药品在胃粘膜防护中的应用。
取明胶10g，吐温0.5ml，加入100ml去离子水，90℃温浴30min至基质完全溶解，另取浓度为10.0mg/ml的贻贝粘蛋白溶液60ml，边搅拌边加入到泡沫基质中，混合均匀后形成贻贝粘蛋白泡沫剂药品，其中贻贝粘蛋白浓度为5mg/g。
收集10例长期服用刺激性药物患者，年龄不限，随机分配至试验组或对照组，试验组在服药前半小时服用贻贝粘蛋白泡沫剂药品，对照组正常服用药物。使用30天后对受试者胃部胃镜检测，试验组5名受试者胃粘膜均正常，对照组5名受试者中2名出现胃粘膜充血症状，2名出现水肿现象，1名出现溃疡症状。表明贻贝粘蛋白泡沫剂药品可用于胃粘膜的防护。
实施例23：贻贝粘蛋白泡沫剂消毒产品在环境污染防护中的应用。
取羟丙基甲基纤维素10g，加入20ml去离子水，0.5ml吐温60，90℃温浴30min至完全溶解得到泡沫剂基质。取浓度为10.0mg/ml的贻贝粘蛋白溶液40ml，制备成贻贝粘蛋白泡沫消毒产品，其中贻贝粘蛋白浓度为2.0mg/ml。同时制备不添加贻贝粘蛋白的泡沫剂基质作为空白组。
收集20例长期接触化学物质刺激受试者，年龄不限，随机分配至试验组或对照组。受试者每天早晚两次使用贻贝粘蛋白泡沫剂消毒产品或空白，将泡沫挤入鼻腔后轻轻按揉至吸收。使用30天后对受试者鼻粘膜进行检测，试验组受试者均无不适现象，对照组有6例出现鼻粘膜充血或水肿现象。证明贻贝粘蛋白泡沫剂消毒产品可用于环境污染下鼻粘膜的防护。

Claims (13)

  1. 贻贝粘蛋白在防护粘膜中的应用。
  2. 根据权利要求1的贻贝粘蛋白应用，其中所述贻贝粘蛋白是来自亚类：mefp1、mefp-2、mefp-3、mefp-4、mefp-5、mefp-6、胶原蛋白pre-COL-P、pre-COL-D、pre-COL-NG、足丝基质蛋白PTMP和DTMP中的一种或几种的混合物。
  3. 根据权利要求1的贻贝粘蛋白应用，其中所述贻贝粘蛋白浓度为0.1-15.0mg/ml。
  4. 根据权利要求1的贻贝粘蛋白应用，其中所述贻贝粘蛋白是以液体剂、凝胶剂或泡沫剂使用。
  5. 根据权利要求1的贻贝粘蛋白应用，其中最终产品中的贻贝粘蛋白是在pH1.0-7.0的范围内，特别是在pH3.0-6.5的范围内。
  6. 根据权利要求1-4中任一项的贻贝粘蛋白应用，其中所述防护粘膜包括防护粘膜免受紫外线、放射线、高温物体、低温物体、海水、环境污染、化学试剂刺激及刺激性药品、食品的损伤。
  7. 贻贝粘蛋白作为活性成分在用于防护粘膜的组合物中的应用，其中所述组合物是以液体剂、凝胶剂或泡沫剂使用。
  8. 根据权利要求7的贻贝粘蛋白应用，其中所述组合物可以通过口服、含化、灌注、滴入、喷雾、吸入、喷涂、腔镜、定向局部缓释、靶向给药、低温或加热方式使用。
  9. 贻贝粘蛋白作为活性成分在用于防护粘膜的药品中的应用。
  10. 贻贝粘蛋白作为活性成分在用于防护粘膜的化妆品中的应用。
  11. 贻贝粘蛋白作为活性成分在用于防护粘膜的消毒产品中的应用。
  12. 贻贝粘蛋白作为活性成分在用于防护粘膜的保健品或食品中的应用。
  13. 贻贝粘蛋白作为活性成分在用于防护皮粘膜的日化产品中的应用。
PCT/CN2016/095362 2015-08-14 2016-08-15 贻贝粘蛋白用于粘膜的防护 WO2017028775A1 (zh)

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