Medical wound surface composite freeze-dried dressing and preparation method thereof

Abstract

The invention discloses a medical wound surface composite freeze-dried dressing and a preparation method thereof, and relates to the field of medicines. The medical wound surface composite freeze-drying dressing comprises a freeze-drying product and a solvent which are independently stored, the freeze-drying product comprises recombinant mussel mucin, recombinant collagen and a freeze-drying protective agent, and the solvent comprises an osmotic pressure regulator and water. By utilizing the synergistic effect of the recombinant collagen and the recombinant mussel mucin, the wound repair effect of the medical wound composite freeze-dried dressing is improved.

Description

Technical Field

[0001] The invention relates to the field of medicine, and in particular to a medical wound composite freeze-dried dressing and a preparation method thereof. Background Art

[0002] With the continuous development of the medical device industry, medical dressing products with recombinant mussel mucin as the main functional ingredient and recombinant collagen as the main functional ingredient are constantly being introduced. Recombinant mussel mucin is rich in positively charged amino acids and dopa groups, which gives it a strong biorepair potential, thus having all-round repair, anti-inflammatory, desensitization, redness reduction and inhibition of melanin deposition. As a biocompatible and biodegradable polymer material, recombinant collagen has broad application prospects in the medical field. From promoting wound healing to tissue regeneration, to drug carriers and beauty skin care, recombinant collagen has brought many benefits to people's health and beauty. However, among the medical device products currently on the market, there are products with recombinant mussel mucin as the core ingredient, and there are also products with recombinant collagen as the core ingredient. There is no product that contains both recombinant mussel mucin and recombinant collagen that can take into account the advantages of both raw materials. At the same time, because the target users are people with small wounds, abrasions, and cuts, whether the product can be sterile is also directly related to the safety of the product. How to ensure that recombinant mussel mucin and recombinant collagen can still maintain their own characteristics and play their roles during the preparation process has become a difficulty restricting the marketing of medical wound composite freeze-dried dressings containing both recombinant mussel mucin and recombinant collagen.

[0003] In the published patent documents, CN111991608A discloses a wound covering and a preparation method thereof, CN112957274A discloses a recombinant collagen freeze-dried ball and a preparation process, CN114917403A discloses a mussel mucin gel and a preparation method and application thereof, CN117247441A discloses a recombinant mussel mucin and an expression system and application thereof, CN118718097A discloses a preparation method of a composite polymer microsphere dermal filler and a dermal filler based thereon, and CN118141969A discloses a multifunctional recombinant mussel mucin medical dressing. The methods disclosed in these patents only utilize the protein characteristics in isolation, without giving play to the potential synergistic effect between these proteins. Summary of the invention

[0004] The purpose of the present invention is to provide a medical wound composite freeze-dried dressing containing both recombinant mussel mucin and recombinant collagen and a preparation method thereof, and to utilize the synergistic effect of recombinant collagen and recombinant mussel mucin to improve the wound repair effect of the medical wound composite freeze-dried dressing.

[0005] The technical solution adopted by the present invention to achieve its purpose is as follows:

[0006] In a first aspect, the present invention provides a medical wound composite freeze-dried dressing, which comprises a freeze-dried product and a solvent stored independently, wherein:

[0007] The freeze-dried product comprises, by weight percentage: 10% to 50% of recombinant mussel mucin, 10% to 50% of recombinant collagen, and 50% to 90% of freeze-drying protective agent;

[0008] The solvent comprises, by weight percentage, 0.9% to 5% of an osmotic pressure regulator and 99.1% to 95.0% of water.

[0009] Furthermore, the lyophilization protective agent is selected from any one or more of trehalose, glucose, sucrose, mannitol, and polyethylene glycol. The amount of the lyophilization protective agent is such that the activity of the protein can be effectively protected and an excellent lyophilized skeleton morphology can be provided.

[0010] Furthermore, the osmotic pressure regulator is selected from any one or more of sodium chloride, glucose, sorbitol, polyethylene glycol, propylene glycol, and mannitol. The osmotic pressure regulator is used in an amount to adjust the osmotic pressure of the composition to isotonic.

[0011] Furthermore, the mass percentage of the lyoprotectant is preferably 80% to 90%.

[0012] Furthermore, the mass percentage of the recombinant mussel mucin is preferably 10% to 20%.

[0013] Furthermore, the mass percentage of the recombinant collagen is preferably 10% to 20%.

[0014] Furthermore, the lyophilized product or the solvent also includes an acid-base regulator, which is used to adjust the pH value within the range of 4 to 7 after the lyophilized product is mixed with the solvent.

[0015] Furthermore, the medical wound composite freeze-dried dressing is of smear type, spray type or application type.

[0016] In a second aspect, the present invention provides a method for preparing a medical wound composite freeze-dried dressing, comprising the following steps:

[0017] Weighing recombinant mussel mucin, recombinant collagen, lyophilization protective agent and water and preparing them, filtering, filling, lyophilizing and sterilizing to obtain a lyophilized product;

[0018] Sodium chloride and water are weighed and prepared, and the solvent is obtained after filtering, filling, plugging and capping, and sterilization.

[0019] Furthermore, the preparation step includes: placing all materials in a sealed container, ultrasonicating for 20 minutes, using ultrasonication to make the solution more fully dissolved to form a uniform and stable solution, and the solid content of the solution during the preparation process is 0.5% to 10%.

[0020] Furthermore, the filtering step includes: passing through a 0.22 um microporous filter membrane.

[0021] Furthermore, the filling step includes: selecting vials of different sizes for filling according to product specifications, with a filling amount of 1 to 10 g.

[0022] Furthermore, the freeze-drying is set up with 8 to 12 total sections, including pre-freezing, primary drying (sublimation) and secondary drying (analysis); wherein the second section to the end of freeze-drying is in the vacuum control section.

[0023] Furthermore, the pre-freezing conditions are:

[0024] Pre-freezing temperature: -35~-50℃, the specific temperature is adjusted according to the material characteristics;

[0025] Pre-freezing time: 2 to 5 hours, to ensure that the material is completely frozen to prevent deformation during the drying process.

[0026] Further, the conditions for the primary drying are:

[0027] Temperature: -35~0℃, divided into several sub-sections, gradually increase the temperature to avoid material damage;

[0028] Time: 8 to 20 hours, adjusted according to material thickness and moisture content.

[0029] Further, the conditions of the secondary drying are:

[0030] Temperature: 0-40℃, divided into several sub-sections, gradually increase the temperature to remove the remaining moisture;

[0031] Time: 10 to 20 hours, adjusted according to material characteristics and drying degree.

[0032] Furthermore, after the freeze drying is completed, the lifting function of the freeze dryer is started under vacuum to perform tamponade. The advantages of tamponade are: tamponade under vacuum ensures that the freeze-dried product is stored in a sterile and oxygen-free environment; during the tamponade, the vacuum degree is strictly controlled to prevent microorganisms and oxygen in the air from entering the product.

[0033] Furthermore, the freeze-dried product is taken out in a whole tray and transported to a bottle feeding, filling and capping integrated machine for capping. The capping is checked by the three-finger method to ensure that there is no looseness and no tilt or curling in appearance.

[0034] Furthermore, the sterilization method is any one of sterilizing filtration, moist heat sterilization, electron beam sterilization, and cobalt 60 irradiation sterilization;

[0035] Among them, sterilization filtration: suitable for heat-sensitive materials, removing microorganisms through filters.

[0036] Moist heat sterilization: Suitable for high temperature resistant materials, using high temperature and high pressure steam for sterilization.

[0037] Electron beam sterilization: Applicable to various materials, using high-energy electron beams to destroy microbial DNA to achieve sterilization effect.

[0038] Cobalt 60 irradiation sterilization: Suitable for radiation-resistant materials, sterilization is carried out using gamma rays emitted by the radioactive isotope cobalt 60.

[0039] Electron beam sterilization is preferred, which has the following advantages: fast sterilization speed and high efficiency; no heating is required, suitable for heat-sensitive materials; no residue after sterilization, no impact on product quality.

[0040] Compared with the prior art, the present invention has the following advantages:

[0041] 1. Innovation of active ingredient combination:

[0042] 1.1. Collaborative repair mechanism:

[0043] Recombinant collagen plays a vital role in wound repair, not only providing the necessary structural support, but also providing a platform for cell migration and proliferation. This support is a key factor in accelerating wound healing.

[0044] Recombinant mussel mucin, with its unique biological activity, promotes cell adhesion and growth, and enhances cell adhesion and activity. The combination of the two forms an efficient synergistic repair mechanism, greatly improving the speed and quality of wound healing.

[0045] 1.2. Integration of multiple functions:

[0046] The multifunctionality of recombinant mussel mucin is reflected in the dopa groups it contains, which can effectively scavenge free radicals and reduce oxidative stress, thereby exerting anti-inflammatory and anti-allergic effects. This is crucial for alleviating inflammation and discomfort around the wound.

[0047] The versatility of collagen is reflected in its ability to replenish collagen lost in the skin and enhance skin elasticity and firmness, which is significantly effective in improving skin appearance and preventing wrinkles.

[0048] 2. Application of freeze-drying technology:

[0049] 2.1.Activity protection:

[0050] Through freeze-drying technology, the dressing of the present invention can effectively remove moisture, thereby avoiding oxidation and degradation of proteins at room temperature, ensuring that the activity of recombinant collagen and recombinant mussel mucin is protected to the greatest extent. The freeze-drying process also allows the product to remain stable without the addition of chemical preservatives, which is of great significance for improving product safety and reducing potential side effects.

[0051] 2.2. Sterility assurance:

[0052] The application of electron beam sterilization technology provides efficient sterility protection for dressings. This sterilization method is not only fast, but also does not cause thermal damage, ensuring that the quality of the dressing is not affected after sterilization.

[0053] The preparation of sterile products greatly reduces the risk of microbial contamination during use and provides consumers with a safer and more reliable choice.

[0054] 3. Multiple application modes to meet different needs:

[0055] The smear type is suitable for small wounds, such as cuts, abrasions, etc. It is convenient, fast and easy to operate.

[0056] The spray type is suitable for areas where it is difficult to apply evenly, such as the face and neck. It is easy to use and provides even coverage.

[0057] The patch type is suitable for large wounds or areas that require long-term coverage, providing continuous action and protection.

[0058] 4. Focus on specific wound surfaces:

[0059] The dressing of the present invention focuses on the repair of non-chronic wounds such as small wounds, abrasions, cuts, and the care of surrounding skin, which makes the dressing more efficient and effective in specific application scenarios. For the repair of skin allergy symptoms, the anti-inflammatory and anti-allergic properties of the dressing of the present invention make it an ideal care choice.

[0060] 5. Potential advantages:

[0061] 5.1. Sustainable repair:

[0062] The gel structure formed by recombinant mussel mucin can form a protective layer on the wound surface and continuously release active ingredients, while the long-term supporting effect of collagen ensures the durability of the repair effect.

[0063] 5.2. Personalized customization:

[0064] According to the needs of different users, the present invention can achieve personalized customization by adjusting the ratio of active ingredients and the formula of lyophilization protectant to meet the special needs of specific user groups.

[0065] In summary, the present invention, through the synergistic effect of recombinant collagen and recombinant mussel mucin, combined with advanced freeze-drying technology and electron beam sterilization technology, not only achieves comprehensive and efficient repair and care functions, but also shows significant advantages in safety, ease of application and market targeting.

[0066] Compared with the prior art, the present invention has the following substantial differences:

[0067] 1. The difference between the present invention and patent CN111991608A

[0068] (1) The main raw materials used are different: the main raw materials used in the present invention are recombinant collagen and recombinant mussel mucin, while the main raw materials used in CN111991608A are ordinary collagen and mussel mucin. The specific differences are shown in the following Tables 1 and 2.

[0069] Table 1 Differences between common mussel mucin and recombinant mussel mucin

[0070]

[0071] Table 2 The difference between ordinary collagen and recombinant collagen

[0072]

[0073] (2) CN111991608A does not mention the protection of activity by freeze-drying technology, but the present invention adds a freeze-drying protective agent to effectively protect the protein. The use of a suitable sterilization process can not only bring into play the synergistic effect of the two proteins, but also make the product sterile and safer.

[0074] (3) CN111991608A has a single application mode (film sheet), while the dressing of the present invention has multiple application modes: smear type, spray type, and application type, and has a wider application range.

[0075] 2. The difference between the present invention and CN112957274A, CN114917403A, CN117247441A, CN118718097A and CN118141969A

[0076] (1) CN112957274A only contains recombinant collagen and chitosan, and does not mention recombinant mussel mucin. CN114917403A only contains mussel-like mucin, without recombinant collagen. CN117247441A only contains recombinant mussel mucin, without recombinant collagen. CN118718097A only contains recombinant mussel mucin, without recombinant collagen. CN118141969A only contains recombinant mussel mucin, without recombinant collagen. None of them can exert the synergistic effect of the two proteins. When the product recombinant collagen and recombinant mussel mucin are used together, the present invention has the following advantages compared to using recombinant collagen alone:

[0077] Enhanced repair ability: Recombinant mussel mucin has excellent film-forming adhesion, rapid repair and stabilization and toughness. This protein is the only known natural protein with a high content of dopa groups, which plays an important role in skin barrier repair. When used in combination with recombinant collagen, these properties can significantly improve the repair effect of damaged skin.

[0078] Improve skin barrier function: Recombinant mussel mucin can promote cell migration and repair and healing, which is particularly important for improving skin barrier function. With the help of recombinant collagen, this promotion effect is more significant, which helps to accelerate the wound healing process.

[0079] Optimized bioactivity: The combination of recombinant mussel mucin and recombinant collagen optimizes the bioactivity of each, which is essential for maintaining skin elasticity and integrity.

[0080] The synergistic effect of recombinant collagen and recombinant mussel mucin shows obvious advantages in improving repair efficiency, enhancing skin barrier function, and optimizing biological activity.

[0081] (2) CN114917403A uses common mussel mucin, while the present invention uses recombinant mussel mucin, which are significantly different from each other.

[0082] (3) The above patent has a single application method, while the dressing of the present invention has multiple application methods: smear type, spray type, and application type, and has a wider application range.

[0083] (4) CN112957274A and CN117247441A do not have a sterilization process, while the present invention uses a suitable sterilization process to make the product sterile, with a wider range of application scenarios and higher safety.

[0084] (5) The pre-freezing time of CN112957274A is 24 hours, while the pre-freezing time of the dressing of the present invention is 2-5 hours, which is more efficient and energy-saving. CN118718097A and CN118141969A do not mention the freeze-drying process, while the present invention adopts the freeze-drying process. The advantages of the freeze-drying process are: 1. Low-temperature drying protects heat-sensitive substances; 2. The freeze-drying process can remove more than 95% to 99% of the water to ensure the stability of the dried product; 3. Since the drying is carried out under vacuum, the easily oxidized substances are protected; 4. The freeze-dried substance has a loose and porous sponge-like structure, which can be quickly dissolved after adding water and almost immediately restore its original properties.

[0085] (6) The sterilization process is different. CN118718097A uses moist heat sterilization, while the sterilization process of the present invention preferably uses electron beam sterilization. Since electron beam sterilization does not rely on heat, it is particularly suitable for the sterilization of heat-sensitive substances. This method can effectively kill bacteria, viruses and other microorganisms without destroying the structure and activity of these substances. Adaptable to a variety of packaging: Whether it is a plastic bag, glass bottle or metal can, the electron beam can penetrate most packaging materials for sterilization, providing more packaging options for the product. Moist heat sterilization requires high temperature and high pressure, which is not only not suitable for heat-sensitive substances, but also limits the packaging form of the product.

[0086] In summary, none of the above five patents mention the synergistic effect of recombinant collagen and recombinant mussel mucin. Although one patent mentions collagen and mussel mucin, the collagen and mussel mucin used are both common types, which are significantly different from the recombinant collagen and recombinant mussel mucin of the present invention. The present invention combines recombinant collagen and recombinant mussel mucin, which can exert the synergistic effect of the two proteins in terms of efficacy, and adopts a new preparation method to obtain an ideal medical wound composite freeze-dried dressing.

[0087] Compared with the prior art, the present invention has the following beneficial effects:

[0088] 1. The composition of the present invention has scientifically selected components and proportions, and the composition can form a protective layer on the wound surface, act as a physical barrier, provide a suitable microenvironment for wound healing, and promote wound healing. It has the advantages of both recombinant mussel viscose dressing and recombinant collagen dressing.

[0089] 2. In the liquid chromatography identification spectrum, the existing sterilization methods, whether electron beam sterilization, cobalt 60 sterilization, or moist heat sterilization, will affect the characteristic peaks of recombinant mussel mucin and recombinant collagen, and sterilization filtration has extremely strict requirements on the production environment and operation of medical device products, high requirements on hardware facilities and equipment, and huge investment. Sterilization process is a difficult point for various companies in the product development process. The composition of the present invention effectively protects the active ingredients of recombinant mussel mucin and recombinant collagen by changing the product dosage form, applying advanced freeze-drying technology and an appropriate proportion of freeze-drying protective agent. At the same time, it effectively solves the problem of sterilization.

[0090] 3. The composition of the present invention is sterilized by electron beam, so that the product can be sterile and applied to wound skin, which is safer, has good biocompatibility and good stability. BRIEF DESCRIPTION OF THE DRAWINGS

[0091] Figure 1 This is a process flow chart for preparing the freeze-dried product of the medical wound composite freeze-dried dressing in Example 1.

[0092] Figure 2 The present invention is a flow chart of the preparation process of the solvent of the medical wound composite freeze-dried dressing of Example 1.

[0093] Figure 3 It is the freeze-drying curve diagram in Example 1. DETAILED DESCRIPTION

[0094] In order to make the various technical features and advantages or technical effects in the above technical solutions of the present invention more obvious and easy to understand, they are described in detail below with reference to the accompanying drawings.

[0095] A medical wound composite freeze-dried dressing prepared by the present invention comprises freeze-dried products and solvents respectively contained in bottles A and B. The specifications of bottles A and B are: A10mg, B2g, A20mg, B2g, A30mg, B2g, A40mg, B2g, A50mg, B2g, A80mg, B2g, A100mg, B2g, A200mg, B2g; A25mg, B5g, A50mg, B5g, A75mg, B5g, A100mg, B5g, A125mg, B5g, A125mg, B5g, A200 mg, B5g, A250mg, B5g, A500mg, B5g; A50mg, B10g, A100mg, B10g, A150mg, B10g, A200mg, B10g, A250mg, B10g, A400mg, B10 g, A500mg, B10g; A100mg, B20g, A200mg, B20g, A300mg, B20g, A400mg, B20g, A500mg, B20g, A800mg, B20g, A1000mg, B20g.

[0096] Embodiment 1:

[0097] The two components of the medical wound composite freeze-dried dressing are bottled in the following specifications: A10mg, B2g, specifically:

[0098] Bottle A freeze-dried product: 1 mg recombinant mussel mucin, 1 mg recombinant collagen, 8 mg trehalose.

[0099] Solution in bottle B: 0.018g sodium chloride, 1.982g water.

[0100] Preparation of freeze-dried product in bottle A: weigh the required amount of recombinant mussel mucin, recombinant collagen, freeze-drying protective agent, and water; prepare, filter, fill, freeze-dry and sterilize.

[0101] Preparation of solution in bottle B: Weigh the required amount of sodium chloride and water; prepare, filter, fill, plug and cap, and sterilize.

[0102] 1. Operation process (taking T-type A10mg and B2g as examples)

[0103] Figure 1 This is a process flow chart for preparing freeze-dried products of medical wound composite freeze-dried dressings. Figure 2 This is a process flow chart for preparing the solvent for medical wound composite freeze-dried dressing. The specific steps are as follows:

[0104] 1. Pre-production inspection:

[0105] A pre-production inspection should be carried out before each production process to confirm that the site is properly cleared, the equipment is in good condition, the measuring instruments have been calibrated and are within the validity period, and the production environment meets the requirements. Only after production is allowed can the production operation of this batch of products be carried out.

[0106] 2. Prepare the ingredients:

[0107] Fill out the material requisition form according to the batch production instruction, collect recombinant mussel mucin, recombinant collagen, trehalose, sodium chloride, vials, freeze-dried brominated butyl rubber stoppers for injection, sterile powder brominated butyl rubber stoppers for injection, and aluminum-plastic combination caps, and collect the raw and auxiliary materials after verification.

[0108] 3. Liquid preparation (Take T-type A10mg, B2g to prepare 1000 bottles as an example)

[0109] 3.1. Preparation of freeze-dried product in bottle A:

[0110] First check whether the weighing instrument is calibrated and within the qualified period, and whether it is zeroed. According to the batch production instructions, weigh the raw materials and excipients, and weigh them double-check. Weigh 70% of the total amount of water for injection into the liquid preparation tank and cool it to below 30°C. Weigh the formula amount of recombinant mussel mucin, recombinant collagen, and trehalose and add them to the liquid preparation tank in turn, stirring for 10 minutes until uniform. Add water for injection to the liquid preparation tank, adjust the volume, stir for 10 minutes, and measure the pH value, which should be between 4.0-7.0.

[0111] 3.2. Preparation of bottle B (solution):

[0112] First check whether the weighing instrument is calibrated and within the qualified period, and whether it is zeroed. According to the batch production instructions, weigh the raw and auxiliary materials, and weigh them double-check. Take 70% of the total formula of water for injection and add it to the batching barrel, and cool it to below 30°C. Weigh the formula amount of sodium chloride in 1% of the total formula of water for injection, stir for 10 minutes to dissolve. Add the sodium chloride solution to the liquid preparation tank, adjust the volume, stir for 10 minutes, and measure the pH value, which should be between 5.0-7.5. Take samples and inspect.

[0113] 4. Filling (taking the preparation of 1000 bottles of T-type A10mg and B2g as an example)

[0114] Before filling, use 75% ethanol solution to disinfect hands and operating table. Bottle A is filled with freeze-dried products; bottle B is filled with solution. Receive intermediate products, vials, freeze-dried bromobutyl rubber stoppers for injection, sterile powder bromobutyl rubber stoppers for injection, and aluminum-plastic combination caps. Wash and dry the vials so that there is no visible foreign matter. Before filling, confirm that the hopper and the filling machine are sealed and connected. After checking that the connections of all parts of the bottle feeding, filling and capping machine are normal, start the machine, first use 75% ethanol solution to circulate and flush the hopper and filling pipeline, and then use injection water to flush the hopper and filling pipeline. At the same time, check whether the machine is running normally. After meeting the requirements, wipe the outer surface of the equipment with 75% ethanol and let it dry naturally. Pour the intermediate liquid to be filled into the hopper, turn on the manual mode and eject it from the filling head, exhaust the gas and excess injection water in the pipeline, and adjust the filling volume: bottle A is filled with 2g of freeze-dried products and bottle B is filled with 2g of solution; after passing the test, formal filling is carried out. Take 5 sample bottles every 15 minutes to check the filling volume. After filling, bottle A will be transferred to the next process for freeze drying. After filling, bottle B will press the rubber stopper onto the vial, put on the aluminum-plastic combination cap, and use the capping machine to cap it.

[0115] 4. Freeze drying:

[0116] Place the filled product (bottle A) into the freeze dryer, insert the temperature probe, and close the door. Before starting the machine, check whether the door and valves are closed to ensure that the cold trap has been defrosted and drained. Start the freeze dryer according to the set formula process, pre-freeze temperature -40℃, and maintain for more than 3 hours; after the product is completely frozen, start the vacuum pump, the vacuum degree drops below 100kpa, and enter the sublimation stage, the sublimation time is 8-20 hours; then enter the secondary drying stage, the temperature must not exceed 35℃, and the time is not less than 10-20 hours. Finally, press the plug and take it out of the box.

[0117] Before starting the freeze dryer, technicians need to set the recipe process parameters according to the product characteristics. The total section is set to 10, which means that there are 10 temperature control sections in this operation procedure; the vacuum control section is set to 2, which means that the vacuum pump is started from the second section, and the vacuum degree in the freeze dryer gradually decreases. The specific process settings are shown in Table 3 below:

[0118] Table 3

[0119]

[0120] The description of primary drying and secondary drying is as follows:

[0121] 1. Primary drying, also called sublimation drying or the main drying stage, aims to remove most of the water from the frozen material, which is present in the material in the form of ice.

[0122] Purpose of primary drying: To remove most of the moisture in the material, usually about 95% of the moisture is removed in this stage.

[0123] Primary drying process: The material is first frozen below the eutectic point to ensure that the water is completely solidified. Subsequently, the material is placed in a vacuum environment and heated to a temperature below the melting point of ice, so that the ice directly changes from solid to gas (sublimation) without passing through the liquid state. At this stage, the temperature needs to be strictly controlled to prevent the sample temperature from exceeding the melting point of ice, resulting in damage to the sample structure or denaturation of the protein.

[0124] 2. Secondary drying, also known as analytical drying or secondary drying stage, is carried out after primary drying to remove the remaining moisture in the material.

[0125] Purpose of secondary drying: To remove the remaining moisture in the material, which is usually bound water or moisture adsorbed in the material structure.

[0126] Secondary drying process: After primary drying, a small amount of moisture remains in the material, which is not easily removed by sublimation. In the secondary drying stage, the temperature is further increased, but usually not high enough to cause thermal damage to the material. By increasing the temperature, the rate of moisture diffusion inside the material can be increased, thereby accelerating the drying process.

[0127] It should be noted that the formulation parameters need to be adjusted when the product specifications, vial sizes, and product loading quantity in the freeze dryer are different. For example, if the specification of bottle A exceeds 200 mg and the freeze dryer is fully loaded, the pre-freezing time and sublimation time need to be appropriately increased.

[0128] Figure 3 It is a freeze-drying curve. The freeze dryer is equipped with a temperature probe. After the bottle A product is placed in the freeze dryer, the probe will record the sample temperature of the dressing throughout the process. At the same time, the freeze dryer will record the parameters of the freeze dryer every 3 minutes during operation, including time, sample temperature, cold trap temperature, set temperature, partition temperature, and vacuum degree. The freeze-drying curve is a curve chart made based on the data automatically recorded by the freeze dryer. The horizontal axis is time and the vertical axis is temperature. The freeze-drying curve chart can clearly show the relationship between sample temperature and time.

[0129] 5. Rolled aluminum-plastic composite cover.

[0130] Before each production operation, the site must be cleared and confirmed. Collect the freeze-dried intermediate products and carefully check the product name, quantity, specification, and batch number. Place the products out of the cabinet on the rotating plate, turn on the rotating button, and make the products move along the track. Turn on the vibration button of the bottle feeding, filling, and capping machine, adjust the speed to check the tightness of the capping and appearance: check the capping by the three-finger method. There should be no looseness, tilting, or curling on the appearance.

[0131] 6. Sterilization

[0132] Solution in bottle B: Moist heat sterilization, sterilization temperature 121℃, time 20min.

[0133] Bottle A freeze-dried product: Contact a qualified sterilization company for electron beam irradiation sterilization. Sterilization conditions: irradiation intensity 8-12kGy.

[0134] 7. Light inspection operation

[0135] Before each production operation, the site must be cleared and confirmed. Check whether the product name, specification, batch number and quantity of the current batch of products are consistent. Check the illumination of the light inspection, and the illumination (not less than 800LX) should meet the requirements. The light inspection operation should be carried out in accordance with the "Standard Operating Procedures for Light Inspection Posts". Lay the products on the light inspection table from left to right in the same direction, check carefully, and pick out products with glass chips, bottles without sample liquid, obvious differences in filling volume, wall adhesion, missing rolling, skirt caps and other unqualified products.

[0136] 8. Outer packaging

[0137] According to the batch packaging instructions, pick up the products to be packaged and check whether the product name, specification, batch number, production date, quantity, number of pieces, etc. are consistent. Check that the packaging materials should not be damaged or damp, and after checking that the materials are consistent with the batch packaging instructions, transport the materials to the packaging room. According to the batch packaging instructions, count the small boxes received and send them to the operator, and print the production batch number, production date, expiration date and other numbers in the specified position of the small box according to the batch packaging instructions. After printing the first small box, hand it over to QA for review. Only after it is correct can you continue to print. The printing should be clear and accurate. After the operation is completed, clear the site, QA will check and issue a clearance certificate. According to the batch packaging instructions, hand the label to the operator, check the quantity and whether it is damaged. According to the batch packaging instructions, hand the large box to the operator, and the operator will stamp the production batch number, production date, expiration date and other numbers in the specified position of the large box according to the batch packaging instructions. After stamping the first large box, hand it over to QA for review. Only after it is correct can you continue to stamp. The stamping should be clear and accurate. According to the batch packaging instructions, the instructions are sent to the operator, who counts the number of instructions for each package according to the packaging specifications and folds them neatly. According to the batch packaging instructions, the products to be packaged are picked up from the intermediate station, and the product name, specifications, quantity, number of pieces, production batch number, production date and expiration date of the product and the packaging material are checked to see if they are consistent. Packing can only be carried out after they are correct.

[0138] 9. Storage

[0139] The workshop staff fills out the inspection request form, and QA takes samples for inspection. After the inspection is qualified, the finished product entry form is filled out, and after the workshop director reviews and signs, it is sent to the warehouse together with the finished product inspection qualified report. The warehouse keeper counts the entry quantity according to the entry form and handles the entry of finished products. Finished product stacking requirements: batches and different varieties must not be mixed.

[0140] 2. Quality Standards

[0141] Intermediate product quality standards

[0142] Bottle A of freeze-dried product should meet the requirements of Table 4 below:

[0143] Table 4

[0144]

[0145] The solution in bottle B should meet the requirements of Table 5 below:

[0146] Table 5

[0147]

[0148] The quality standards of finished products meet the requirements of Table 6 below:

[0149] Table 6

[0150]

[0151] Embodiment 2:

[0152] The two components of the medical wound composite freeze-dried dressing are bottled in the following specifications: A20mg, B2g, specifically:

[0153] Bottle A freeze-dried product: 2 mg recombinant mussel mucin, 2 mg recombinant collagen, 16 mg trehalose.

[0154] Solution in bottle B: 0.018g sodium chloride, 1.982g water.

[0155] Preparation of freeze-dried product in bottle A: weigh the required amount of recombinant mussel mucin, recombinant collagen, freeze-drying protective agent, and water; prepare, filter, fill, freeze-dry and sterilize.

[0156] Preparation of solvent in bottle B: Weigh the required amount of sodium chloride and water; prepare, filter, fill, plug and cap, and sterilize.

[0157] Embodiment 3:

[0158] The two components of the medical wound composite freeze-dried dressing are bottled in the following specifications: A30mg, B2g, specifically:

[0159] Bottle A freeze-dried product: 3 mg recombinant mussel mucin, 3 mg recombinant collagen, 24 mg trehalose.

[0160] Solution in bottle B: 0.018g sodium chloride, 1.982g water.

[0161] Preparation of freeze-dried product in bottle A: weigh the required amount of recombinant mussel mucin, recombinant collagen, freeze-drying protective agent, and water; prepare, filter, fill, freeze-dry and sterilize.

[0162] Preparation of solvent in bottle B: Weigh the required amount of sodium chloride and water; prepare, filter, fill, plug and cap, and sterilize.

[0163] Embodiment 4:

[0164] The two components of the medical wound composite freeze-dried dressing are bottled in the following specifications: A40mg, B2g, specifically:

[0165] Bottle A freeze-dried product: 4 mg recombinant mussel mucin, 4 mg recombinant collagen, 32 mg trehalose.

[0166] Solution in bottle B: 0.018g sodium chloride, 1.982g water.

[0167] Preparation of freeze-dried product in bottle A: weigh the required amount of recombinant mussel mucin, recombinant collagen, freeze-drying protective agent, and water; prepare, filter, fill, freeze-dry and sterilize.

[0168] Preparation of solvent in bottle B: Weigh the required amount of sodium chloride and water; prepare, filter, fill, plug and cap, and sterilize.

[0169] Embodiment 5:

[0170] The two components of the medical wound composite freeze-dried dressing are bottled in the following specifications: A50mg, B2g, specifically:

[0171] Bottle A freeze-dried product: 5 mg recombinant mussel mucin, 5 mg recombinant collagen, 40 mg trehalose.

[0172] Solution in bottle B: 0.018g sodium chloride, 1.982g water.

[0173] Preparation of freeze-dried product in bottle A: weigh the required amount of recombinant mussel mucin, recombinant collagen, freeze-drying protective agent, and water; prepare, filter, fill, freeze-dry and sterilize.

[0174] Preparation of solvent in bottle B: Weigh the required amount of sodium chloride and water; prepare, filter, fill, plug and cap, and sterilize.

[0175] Embodiment 6:

[0176] The two components of the medical wound composite freeze-dried dressing are bottled in the following specifications: A80mg, B2g, specifically:

[0177] Bottle A freeze-dried product: 8 mg recombinant mussel mucin, 8 mg recombinant collagen, 72 mg trehalose.

[0178] Solution in bottle B: 0.018g sodium chloride, 1.982g water.

[0179] Preparation of freeze-dried product in bottle A: weigh the required amount of recombinant mussel mucin, recombinant collagen, freeze-drying protective agent, and water; prepare, filter, fill, freeze-dry and sterilize.

[0180] Preparation of solvent in bottle B: Weigh the required amount of sodium chloride and water; prepare, filter, fill, plug and cap, and sterilize.

[0181] Embodiment 7:

[0182] The two components of the medical wound composite freeze-dried dressing are bottled in the following specifications: A100mg, B2g, specifically:

[0183] Bottle A freeze-dried product: recombinant mussel mucin 10 mg, recombinant collagen 10 mg, trehalose 80 mg.

[0184] Solution in bottle B: 0.018g sodium chloride, 1.982g water.

[0185] Preparation of freeze-dried product in bottle A: weigh the required amount of recombinant mussel mucin, recombinant collagen, freeze-drying protective agent, and water; prepare, filter, fill, freeze-dry and sterilize.

[0186] Preparation of solution in bottle B: Weigh the required amount of sodium chloride and water; prepare, filter, fill, plug and cap, and sterilize.

[0187] Embodiment 8:

[0188] The two components of the medical wound composite freeze-dried dressing are bottled in the following specifications: A200mg, B2g, specifically:

[0189] Bottle A freeze-dried product: 20 mg recombinant mussel mucin, 20 mg recombinant collagen, 160 mg trehalose.

[0190] Solution in bottle B: 0.018g sodium chloride, 1.982g water.

[0191] Preparation of freeze-dried product in bottle A: weigh the required amounts of recombinant mussel mucin, recombinant collagen, freeze-drying protective agent, and water; prepare, filter, fill, freeze-dry, and sterilize to obtain the product.

[0192] Preparation of solution in bottle B: Weigh the required amount of sodium chloride and water; prepare, filter, fill, plug and cap, and sterilize.

[0193] Embodiment 9:

[0194] The two components of the medical wound composite freeze-dried dressing are bottled in specifications of A500mg and B5g, specifically:

[0195] Bottle A freeze-dried product: 50 mg recombinant mussel mucin, 50 mg recombinant collagen, 400 mg trehalose.

[0196] Solution in bottle B: 0.045g sodium chloride, 4.955g water.

[0197] Preparation of freeze-dried product in bottle A: weigh the required amount of recombinant mussel mucin, recombinant collagen, freeze-drying protective agent, and water; prepare, filter, fill, freeze-dry and sterilize.

[0198] Preparation of solution in bottle B: Weigh the required amount of sodium chloride and water; prepare, filter, fill, plug and cap, and sterilize.

[0199] How to use the medical wound composite freeze-dried dressing:

[0200] The types of use of medical wound composite freeze-dried dressings are divided into smear type, spray type, and dressing type. The use methods are as follows:

[0201] Smear type: Clean the affected skin. Open the package, pour the solution in bottle B into the freeze-dried product in bottle A, cover with a rubber stopper, mix well, and apply the liquid directly to the affected area.

[0202] Spray type: Clean the affected skin. Open the package, pour the solution in bottle B into the freeze-dried product in bottle A, cover with a rubber stopper and mix well, then remove the rubber stopper, install the matching spray head on the bottle, press the spray pump head, and spray on the affected area.

[0203] Patching type: Clean the affected skin. Open the package, pour the solution in bottle B into the freeze-dried product in bottle A, cover with a rubber stopper and mix evenly, then remove the rubber stopper, soak the matching non-woven fabric completely with the mixed solution, and apply it to the affected area.

[0204] Performance test of medical wound composite freeze-dried dressing:

[0205] 1. Formula of medical wound composite freeze-dried dressing (this product)

[0206] The formula of freeze-dried product in bottle A is shown in Table 7:

[0207] Table 7

[0208]

[0209] The solution formula of bottle B is shown in Table 8:

[0210] Table 8

[0211]

[0212] 2. Film formation test research:

[0213] In the experiment, bottles A10 and B10 of medical recombinant mussel mucin and recombinant collagen freeze-dried dressings were taken and spread on a glass plate. They were placed at a temperature of 65°C and dried for 4 hours. A protective film was formed in the culture dish.

[0214] 3. Physical and chemical properties research:

[0215] The main performance indicators of the product are shown in Table 9:

[0216] Table 9

[0217]

[0218]

[0219] 4. Biological characteristics research:

[0220] 1. Evaluation basis and methods: Biocompatibility evaluation is based on GB / T 16886.1-2022 Biological Evaluation of Medical Devices Part 1: Risk Management Evaluation and Testing. Description of the materials used in the product and the nature of contact with the human body.

[0221] The manufacturing materials are: recombinant mussel mucin, recombinant collagen, trehalose, sodium chloride, and water for injection.

[0222] The ingredients that come into direct contact with the patient or user's body are: recombinant mussel mucin, recombinant collagen, trehalose, sodium chloride, and water for injection.

[0223] The results of the biological tests are shown in Table 10:

[0224] Table 10

[0225] Test Project Evaluation Criteria result Cytotoxicity assay The cell viability of the test product is ≥75%, with no potential toxic effects No potential toxic effects, qualified Animal irritation test Should not be greater than 0.4 No irritation, qualified Skin sensitization test Should not be greater than level 1 The stimulation index is 0, which is qualified. Materials-mediated pyrogenicity No material-mediated pyrogenicity No material-mediated pyrogenicity, qualified Acute systemic toxicity No acute systemic toxicity No acute systemic toxicity, qualified

[0226] Although the present invention has been disclosed as above by way of embodiments, it is not intended to limit the present invention. Appropriate modifications or equivalent substitutions of the technical solutions of the present invention made by ordinary technicians in the field should all be included in the protection scope of the present invention. The protection scope of the present invention shall be based on what is defined in the claims.

Claims

1. A medical wound composite freeze-dried dressing, comprising a freeze-dried product and a solvent stored independently, wherein: The freeze-dried product comprises, by weight percentage: 10% to 50% of recombinant mussel mucin, 10% to 50% of recombinant collagen, and 50% to 90% of freeze-drying protective agent; The solvent comprises, by weight percentage, 0.9% to 5% of an osmotic pressure regulator and 99.1% to 95.0% of water.

2. The medical wound composite freeze-dried dressing according to claim 1, characterized in that: The lyophilization protective agent is selected from any one or more of trehalose, glucose, sucrose, mannitol, and polyethylene glycol; the osmotic pressure regulator is selected from any one or more of sodium chloride, glucose, sorbitol, polyethylene glycol, propylene glycol, and mannitol.

3. The medical wound composite freeze-dried dressing according to claim 1 or 2, characterized in that: The mass percentage of the lyophilization protectant is 80% to 90%, the mass percentage of the recombinant mussel adhesive protein is 10% to 20%, and the mass percentage of the recombinant collagen is 10% to 20%.

4. The medical wound composite freeze-dried dressing according to claim 1, characterized in that: The freeze-dried product or solvent also includes an acid-base regulator, which is used to adjust the pH to within the range of 4 to 7 after the freeze-dried product is mixed with the solvent.

5. The medical wound composite freeze-dried dressing according to claim 1, characterized in that: The medical wound composite freeze-dried dressing is of smear type, spray type or application type.

6. A method for preparing the medical wound composite freeze-dried dressing according to any one of claims 1 to 5, characterized in that: The following steps are involved: Weighing recombinant mussel mucin, recombinant collagen, lyophilization protective agent and water and preparing them, filtering, filling, lyophilizing and sterilizing to obtain a lyophilized product; Sodium chloride and water are weighed and prepared, and the solvent is obtained after filtering, filling, plugging and capping, and sterilization.

7. The preparation method according to claim 6, characterized in that: The preparation steps include: placing all materials in a sealed container, ultrasonicating for 20 minutes, using ultrasonication to make the solution more fully dissolved to form a uniform and stable solution, and the solid content of the solution during the preparation process is 0.5% to 10%; The filtering step comprises: passing through a 0.22 um microporous filter membrane; The filling step comprises: selecting vials of different sizes for filling according to the specifications of the product, with the filling amount being 1 to 10 g.

8. The preparation method according to claim 6, characterized in that: The freeze-drying setting has 8 to 12 total sections, including pre-freezing, primary drying and secondary drying; the second section to the end of freeze-drying is in the vacuum control section; The pre-freezing conditions are: pre-freezing temperature: -35 to -50°C, pre-freezing time: 2 to 5 hours; The conditions of the primary drying are: temperature of -35 to 0°C, divided into several sub-sections, and gradually increasing the temperature; and time of 8 to 20 hours. The secondary drying conditions are: temperature 0-40° C., divided into several sub-sections, gradually increasing temperature; time 10-20 hours.

9. The preparation method according to claim 6, characterized in that: After the freeze drying is completed, the lifting function of the freeze dryer is started under vacuum to perform plugging; the entire tray of the freeze dried product is taken out and transported to the bottle feeding, filling and capping integrated machine for capping.

10. The preparation method according to claim 6, characterized in that: The sterilization method is any one of sterilizing filtration, moist heat sterilization, electron beam sterilization, and cobalt 60 irradiation sterilization.

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