Multi-target complex egg yolk globulin against respiratory infection and preparation method thereof

Abstract

The application discloses a multi-target composite egg yolk globulin against respiratory tract infection and a preparation method thereof, relates to the technical field of biological medicine, and comprises multi-target composite chicken-derived induced-natural composite antibody, food-grade natural immune enhancing component, sodium chloride and purified water. In the preparation, an isotonic aqueous solution is first prepared, the pre-diluted natural immune enhancing component is sequentially added, the antibody powder is gently dissolved under low temperature, and then the product is filled after filtration. The application can realize the synergistic effect of passive neutralization and active mucosal immune repair, improve the activity retention rate of the antibody after atomization, prolong the local action time of the active ingredient, has good broad-spectrum and high safety, and is suitable for the prevention and adjuvant treatment of respiratory tract infection.

Description

Technical Field

[0001] This invention relates to the field of biomedical technology, specifically to a multi-target complex egg yolk globulin for combating respiratory infections (including colds, asthma, bronchitis, white lung, rhinitis, etc.) and its preparation method. Background Technology

[0002] Respiratory infections are among the most common types of disease globally, posing a persistent threat to public health. Lower respiratory tract infections remain a significant source of morbidity and mortality worldwide. The pathogen spectrum is complex, encompassing multiple pathogenic factors including viruses, bacteria, and allergens, often presenting as mixed infections clinically. Given this situation, the development of safe and broad-spectrum respiratory protective products is a real need.

[0003] Currently, there are several patents published regarding the application of egg yolk antibodies (IgY) in the prevention and treatment of respiratory infections. Patent publication number CN1477128A discloses a method for preparing specific IgY for respiratory infections and its combined formulations. This patent employs a single antigen immunization strategy to prepare specific antibodies, focusing on pathogen neutralization. Patent publication number CN108992669B discloses a composite egg yolk antibody composition for preventing and treating respiratory infections, a nebulized inhalation solution, and its preparation process and application. This patent directly formulates the composite egg yolk antibody into a nebulized inhalation agent, but its technical solution remains limited to the single pathway of antibody neutralization. None of the above-mentioned prior art involves introducing active immune regulation or mucosal repair functional components into the antibody preparation, nor does it systematically optimize the protection of protein activity of egg yolk antibodies during nebulization.

[0004] Existing egg yolk preparations only provide exogenous antibodies to neutralize pathogens and cannot actively promote the host's own mucosal immune repair. After viral infection, the respiratory mucosal barrier is damaged, and passive neutralization alone is insufficient to accelerate mucosal repair and host immune reconstruction, resulting in a short protective period and limited effectiveness. Furthermore, egg yolk antibodies are easily denatured and inactivated by shear forces during ultrasonic or micro-mesh nebulization; current technologies do not incorporate targeted protective measures, leading to low antibody activity retention rates after nebulization.

[0005] Therefore, how to overcome the limitations of a single passive immunization pathway and simultaneously provide exogenous antibody neutralization while also ensuring mucosal barrier repair and antibody activity protection during the atomization process is an unsolved technical problem in this field. Summary of the Invention

[0006] The purpose of this invention is to overcome the shortcomings of the prior art and provide a multi-target complex egg yolk globulin for resisting respiratory tract infections and its preparation method. By co-dissolving the multi-target complex chicken-derived inducible-natural complex antibody with food-grade natural immune-enhancing components in an isotonic aqueous matrix, and combining low-temperature mild dissolution and aseptic filtration preparation process, a dual-pathway synergistic effect of neutralizing exogenous pathogens and repairing endogenous mucosal immunity is achieved. At the same time, the antibody activity during the nebulization process is effectively protected, thereby improving the protective and adjuvant therapeutic effects of respiratory tract infections.

[0007] To solve the above-mentioned technical problems, the present invention provides the following technical solution: On one hand, a multi-target complex egg yolk globulin for resisting respiratory infections, wherein the multi-target complex egg yolk globulin is a broad-spectrum anti-respiratory pathogen multi-target chicken-derived inducible-natural complex antibody nebulization inhalation composition containing food-grade natural immune-enhancing components, the composition comprising the following components:

[0008] A multi-target complex chicken-derived inducible-natural complex antibody with a content of 0.5%-2.5% w / v, wherein the multi-target complex chicken-derived inducible-natural complex antibody contains specific antibodies against viral antigens and bacterial antigens;

[0009] At least one food-grade natural immune-enhancing ingredient, with a content of 0.01%-1.0% v / v, wherein the food-grade natural immune-enhancing ingredient is selected from one or more of plant-derived immunomodulatory polysaccharides, food-grade mucosal repair factors, and food-grade antioxidant and anti-inflammatory plant extracts;

[0010] Sodium chloride is used as an isotonic regulator at a concentration of 0.9% w / v;

[0011] The remainder is purified water;

[0012] The multi-target complex chicken-derived inducible-natural complex antibody and the food-grade natural immune-enhancing ingredient are co-soluble in the same aqueous matrix and act synchronously on the respiratory mucosa after being atomized by ultrasound or micro-mesh, achieving a dual-pathway synergistic effect of passive neutralization and active mucosal immune repair and antibody synthesis promotion; and the food-grade natural immune-enhancing ingredient does not include steviosides, fruit juices, menthol and other substances that only provide taste or physical relief and have no positive antibody effect.

[0013] This composition co-dissolves multi-target complex egg yolk globulin with food-grade natural immune-enhancing ingredients in an isotonic aqueous solution, and exerts both exogenous neutralization and endogenous immune repair effects after nebulization. The antibody titer retention rate after nebulization reached 92.5%, which is 12.5 percentage points higher than the control group without natural ingredients (approximately 80%), confirming that the synergistic effect of the dual pathways has a quantifiable gain.

[0014] Furthermore, the plant-derived immunomodulatory polysaccharides include Polygonatum odoratum extract, which is pre-prepared as a 50-200 times diluted standard solution, and the amount added per 5 mL of composition is 0.01-0.15 mL;

[0015] The food-grade mucosal repair factor includes sodium hyaluronate, which is pre-prepared as a 150-300 times diluted standard solution, and the amount added per 5 mL of composition is 0.01-0.08 mL.

[0016] Adding 0.01-0.15 mL of Polygonatum odoratum extract and 0.01-0.08 mL of sodium hyaluronate per 5 mL can respectively achieve intestinal-lung immune regulation and prolong mucosal retention. Sodium hyaluronate extended the half-life of antibodies on porcine tracheal mucosa from 42 minutes to 55 minutes, an extension of approximately 31%, effectively improving the local action duration of the active ingredients.

[0017] Furthermore, the food-grade antioxidant and anti-inflammatory plant extract is selected from one or more of perilla extract, lily extract, monk fruit extract, and honeysuckle extract, and is pre-prepared as a 100-200 times diluted standard solution, with an addition amount of 0.02-0.10 mL per 5 mL of the composition;

[0018] The composition also contains one or more auxiliary stabilizing ingredients selected from erythritol solution, food-grade glycerin, and food-grade lactic acid.

[0019] Extracts from plants such as perilla, lily, or monk fruit, added at 0.02-0.10 mL per 5 mL, can inhibit inflammatory pathways and reduce antibody oxidative consumption. Extracts from Polygonatum odoratum increased the virus neutralizing titer from 1:6400 to 1:7680, a relative increase of 20%, while taste modifiers such as steviol glycosides did not have this effect, demonstrating that the selected extracts possess a clear auxiliary immune function.

[0020] Furthermore, in a single-preparation composition, the types of functional ingredients in the food-grade natural immune-enhancing ingredients do not exceed two, and erythritol, glycerin, and other excipients are not included, as these are optional excipients.

[0021] The number of functional natural ingredients in a single formulation does not exceed two, avoiding the problem of each ingredient falling below its effective threshold when multiple components are added at low concentrations. The T0 top-tier formula contains only two ingredients: Polygonatum odoratum extract and sodium hyaluronate. After atomization, the potency retention rate and mucosal retention time are significantly better than the control group with a single ingredient or no natural ingredients, ensuring accurate dosage and controllable efficacy.

[0022] Furthermore, the viral antigen set includes at least three of the following: influenza virus antigen, novel coronavirus antigen, respiratory syncytial virus antigen, adenovirus antigen, and rhinovirus antigen.

[0023] The bacterial antigen group includes at least two of the following: Streptococcus pneumoniae antigen, Haemophilus influenzae antigen, group A type B hemolytic streptococcal antigen, and Staphylococcus aureus antigen.

[0024] The antibodies are prepared simultaneously targeting at least three viral antigens, two bacterial antigens, and one allergen antigen, covering common pathogens such as influenza virus, coronavirus, Streptococcus pneumoniae, and house dust mites. The combined immunization regimen utilizes conserved regions such as the HA2 fusion peptide, the S2 conserved region, and the F protein conserved epitope, resulting in antibodies that can cross-neutralize different subtypes, making them suitable for mixed and secondary infections.

[0025] Furthermore, the viral antigen set includes conserved viral protein region antigens, which are selected from one or more of the following: influenza virus HA2 fusion peptide, novel coronavirus S2 conserved region, and respiratory syncytial virus F protein conserved epitope.

[0026] Using influenza virus HA2 fusion peptide, SARS-CoV-2 S2 conserved region, or respiratory syncytial virus F protein conserved epitope as immunogens, and leveraging the high response of the avian immune system to mammalian conserved antigens, the resulting antibodies can recognize common epitopes among different viral subtypes. This method can reduce the risk of inactivation due to viral surface protein mutations and extend the effective window period of the product.

[0027] Furthermore, the composition has a pH of 5.5-7.0, an osmotic pressure of 280-320 mOsm / kg, and no visible particulate matter after filtration through a 0.22 μm microporous membrane.

[0028] The composition, with a pH controlled at 5.5-7.0 and an osmotic pressure of 280-320 mOsm / kg, and sterilized via a 0.22 μm filter membrane, maintains the native conformation of the antibody protein while avoiding osmotic irritation to the respiratory mucosa. The finished product has a pH of 6.2 and an osmotic pressure of 298 mOsm / kg, and remains transparent and clear after dissolution and filtration at 4°C, meeting the basic requirements for nebulized inhalation formulations.

[0029] On the other hand, the preparation method of multi-target complex egg yolk globulin for anti-respiratory tract infection is applicable to multi-target complex egg yolk globulin for anti-respiratory tract infection, and the method includes the following steps:

[0030] S1, Dissolve sodium chloride in purified water to prepare a 0.9% isotonic aqueous solution;

[0031] S2, add one or two pre-diluted food-grade natural immune-enhancing ingredients to the solution of S1 in sequence, and stir well;

[0032] S3, add multi-target complex chicken-derived inducible-natural complex antibody powder to the mixture of S2, and gently stir at a low temperature of 2-8℃ until completely dissolved;

[0033] S4, add purified water to the target volume, filter through a 0.22μm microporous membrane, fill and seal to obtain the product.

[0034] This preparation method first prepares an isotonic base solution, then adds pre-diluted natural components, and finally gently dissolves the antibody powder at 2-8°C, effectively reducing protein denaturation. The composition prepared using this method achieved a titer retention rate of 92.5% after atomization, while the unoptimized simple antibody control only achieved 80%, indicating that low temperature, gentle operation, and the addition of natural components collectively protected antibody activity.

[0035] Compared with existing technologies, this multi-target complex egg yolk globulin for resisting respiratory infections and its preparation method have the following beneficial effects:

[0036] I. This invention co-dissolves a multi-target, chicken-derived inducible-natural complex antibody with a food-grade natural immune-enhancing component in the same aqueous matrix. After nebulization, the two components act simultaneously on the respiratory mucosa. While providing exogenous antibodies to neutralize various respiratory pathogens, it actively promotes the repair of the respiratory mucosal barrier and the reconstruction of the host's own immunity. At the same time, it effectively protects the natural conformation of the antibody during the nebulization process, thereby playing a preventive and adjunctive therapeutic role against various respiratory diseases such as colds, asthma, bronchitis, white lung, and rhinitis. It avoids protein denaturation and inactivation caused by shear force, solving the common industry problems of short protection cycle, limited effect, and low antibody activity retention rate after nebulization in existing single passive immune pathways.

[0037] II. This invention uses multiple antigens, including conserved viral protein region antigens, to immunize laying hens to prepare composite antibodies. These antibodies can cover a variety of common respiratory viruses, bacteria, and allergens, making them suitable for mixed and secondary infection scenarios. They reduce the risk of product failure caused by viral surface protein mutations. At the same time, food-grade natural immune-enhancing ingredients are selected, and the types of functional ingredients in each formulation are strictly controlled to ensure the safety and efficacy stability of the product. The preparation process is simple to operate, the conditions are mild, and it is easy to achieve large-scale industrial production.

[0038] Other advantages, objectives and features of the invention will be set forth in part in the description which follows, and in part will be apparent to those skilled in the art from the following examination or study, or may be learned from the practice of the invention. Attached Figure Description

[0039] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the accompanying drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are merely some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without any creative effort.

[0040] Figure 1 This is a schematic diagram of the overall technical architecture of the present invention;

[0041] Figure 2 This is a schematic diagram of the atomization delivery process of the present invention;

[0042] Figure 3 This is a schematic diagram of the dual-pathway synergistic mechanism of the present invention. Detailed Implementation

[0043] To further illustrate the technical means and effects of the present invention in achieving its intended purpose, the following detailed description of the specific implementation methods, structures, features, and effects of the present invention, in conjunction with the accompanying drawings and preferred embodiments, is provided below.

[0044] like Figure 1 As shown, the multi-target complex egg yolk globulin for combating respiratory infections provided by this invention has an overall technical architecture consisting of a passive neutralizing layer and an active gain layer. The passive neutralizing layer contains chicken-derived inducible-natural complex antibodies targeting three antigen groups: viruses, bacteria, and allergens. The active gain layer contains food-grade natural immune-enhancing components. Both components are co-soluble in an isotonic aqueous solution and act simultaneously on the respiratory mucosa after nebulization.

[0045] The following representative embodiment provides a complete description of the composition, preparation method, and application effects of the present invention. In this embodiment, all raw materials are commercially available food-grade or pharmaceutical-grade products, and all diluents are pre-prepared according to a uniform standard before use.

[0046] This embodiment first prepares a combined immunogenic composition for immunizing laying hens. The specific steps are as follows:

[0047] Take 40 mg each of influenza A H1N1 subtype HA2 fusion peptide antigen, SARS-CoV-2 S2 conserved region antigen, and respiratory syncytial virus F protein conserved epitope antigen, and mix them as the viral antigen group. Take 20 mg each of Streptococcus pneumoniae surface protein A antigen and Haemophilus influenzae outer membrane protein P6 antigen, and mix them as the bacterial antigen group. Mix the above antigens with complete Freund's adjuvant at a weight ratio of 1:1, and emulsify them at 12,000 rpm for 5 minutes using a high-speed homogenizer to form a water-in-oil emulsion, which is the combined immunogenic composition.

[0048] Sixty healthy 20-week-old white Leghorn laying hens were selected and housed in an SPF-grade animal facility. Each hen received a primary immunization consisting of 0.5 mL of the combined immunogenic antigen composition injected at multiple sites in the breast muscle. A second immunization was administered 14 days after the primary immunization, using a 1:1 mixture of incomplete Freund's adjuvant and the antigen, with the same injection dose and site. A third immunization was administered 28 days after the primary immunization, using an aluminum adjuvant at a dose of 0.3 mL.

[0049] Starting 21 days after the initial immunization, two eggs were collected weekly. The titers of antibodies against influenza virus HA2, coronavirus S2, pneumococcus, and house dust mites were measured using indirect ELISA. High-immunity eggs were collected when the anti-influenza virus antibody titer reached 1:32000 or higher, and the titers of the other antibodies were all at least 1:16000. The collection period was from day 7 to day 60 after the third immunization, with a total of approximately 800 high-immunity eggs collected.

[0050] The collected high-immunity eggs were refrigerated at 4°C. Before use, the eggshell surface was wiped with 75% alcohol. The yolks were manually separated from the shells, and the yolk liquid was collected. 1000g of yolk liquid was added to 8L of sterile deionized water at a mass-to-volume ratio of 1:8, and the pH was adjusted to 5.2 with 1mol / L hydrochloric acid. The mixture was placed in a 4°C freezer and stirred at low speed of 80rpm for 2 hours, then allowed to stand overnight.

[0051] The following day, the mixture was centrifuged at 8000g for 30 minutes after settling to remove precipitated lipoproteins, and the supernatant was collected. Carrageenan powder was added to the supernatant to a final concentration of 0.3%, and the mixture was stirred slowly for 20 minutes. The mixture was then allowed to stand for 1 hour to allow complete flocculation and precipitation. A plate and frame filter press with 0.45μm filter paper was used to collect the clarified filtrate.

[0052] The clarified filtrate was concentrated using a regenerated cellulose ultrafiltration membrane with a molecular weight cutoff of 100 kDa. The feed pressure was controlled at 0.2 MPa, and the permeate flow rate was approximately 120 mL / min. When the concentrate volume was reduced to one-tenth of its original volume, washing with an equal volume of purified water was initiated, and the mixture was washed three times. The concentrate was then collected.

[0053] A preservative was added to the concentrate. The preservative consisted of trehalose, sorbitol, and maltodextrin mixed in a mass ratio of 1:1:2, and the amount added was 10% of the solid content of the concentrate. After complete dissolution, the concentrate was freeze-dried. The freeze-drying conditions were as follows: pre-freezing temperature -40℃ for 4 hours; primary drying temperature -20℃ and pressure 15Pa for 24 hours; secondary drying temperature 25℃ and pressure 5Pa for 8 hours. The resulting white to pale yellow powder was the multi-target complex egg yolk globulin powder. Its protein content was determined to be 86.3 g / 100 g, globulin content 52.1 g / 100 g, γ-globulin content 22.4%, and moisture content 5.2 g / 100 g.

[0054] All natural ingredients must be pre-diluted into a standard solution before use to precisely control the amount added. The specific preparation method is as follows:

[0055] Polygonatum odoratum extract: Take commercially available Polygonatum odoratum extract stock solution and dilute it 100 times with purified water. That is, take 1 mL of stock solution and add purified water to 100 mL. Mix well and refrigerate at 2-8℃ for later use.

[0056] Sodium hyaluronate solution: Take sodium hyaluronate powder, dissolve it in purified water and dilute it 200 times, that is, take 0.1g of sodium hyaluronate and add purified water to 200mL. After it is fully dissolved, refrigerate it at 2-8℃ for later use.

[0057] Lily extract: Take commercially available lily extract stock solution and dilute it 100 times with purified water, using the same method as above.

[0058] Monk fruit extract: Take commercially available monk fruit extract stock solution and dilute it 100 times with purified water.

[0059] Perilla extract: Take commercially available perilla extract stock solution and dilute it 200 times with purified water.

[0060] Erythritol solution: Take erythritol powder, dissolve it in purified water and dilute it 100 times, that is, take 1g of erythritol and add purified water to 100mL.

[0061] Food-grade glycerin solution: Take food-grade glycerin and dilute it 150 times with purified water, that is, take 0.67 mL of glycerin and add purified water to 100 mL.

[0062] The following is a detailed explanation using the 10mL T0 premium formula as an example. This formula contains two natural immune-enhancing ingredients: Polygonatum odoratum polysaccharide and sodium hyaluronate.

[0063] Weigh 0.09 g of sodium chloride and place it in a 50 mL beaker. Add about 8 mL of purified water and stir until completely dissolved to obtain a 0.9% isotonic sodium chloride aqueous solution. Add 0.10 mL of pre-diluted Polygonatum odoratum extract and 0.05 mL of sodium hyaluronate solution to this solution in sequence. Stir at 200 rpm for 1 minute with a magnetic stirrer to mix evenly.

[0064] Weigh 0.10 g of the prepared egg yolk globulin powder and slowly add it to the mixture. Place the beaker in a 4°C water bath and gently stir at 100 rpm for about 10 minutes until the powder is completely dissolved. Avoid vigorous shaking or high-speed stirring to prevent antibody protein denaturation.

[0065] Add purified water to a total volume of 10 mL, filter and sterilize using a 0.22 μm polyethersulfone microporous membrane, and collect the filtrate in a sterile container. Fill each disposable medical nebulizer cup with 10 mL of filtrate, seal and label, and store at 2-8°C, indicating that it should be prepared and used immediately and should not be kept overnight.

[0066] The finished product is a colorless to slightly yellow transparent liquid. The pH value is 6.2 as measured by a pH meter, and the osmotic pressure is 298 mOsm / kg as measured by an osmometer, which meets the isotonic requirements for respiratory mucosal administration.

[0067] The nebulized inhalation composition is used as follows:

[0068] To use, connect the nebulizer cup containing the composition to a micro-mesh nebulizer (such as a medical compressor nebulizer or vibrating mesh nebulizer). Turn on the nebulizer, set the nebulization rate to 0.3 mL / min, and the nebulization time for each session to 8-10 minutes, ensuring complete atomization of the 10 mL composition. The user wears a mask or mouthpiece and inhales the mist through the mouth and nose. Use 1-2 times daily for 5-7 consecutive days as a prevention or adjunctive treatment cycle. It is suitable for the adjunctive improvement of respiratory symptoms such as colds, asthma, bronchitis, white lung, and rhinitis. For sensitive individuals, infants, or asthma patients, halve the amount of all natural immune-enhancing ingredients; that is, replace the Polygonatum odoratum extract with 0.05 mL and the sodium hyaluronate solution with 0.025 mL, while keeping the other components unchanged.

[0069] Antibody titer retention rate determination before and after nebulization:

[0070] The above-mentioned T0 top-grade formulation composition was used as the experimental group. A control composition was prepared, containing only 0.9% sodium chloride and 1.0% multi-target complex egg yolk globulin powder, but without Polygonatum odoratum extract and sodium hyaluronate. Both compositions were nebulized for 8 minutes using the same micro-mesh nebulizer. Liquid samples were collected before and after nebulization, and the anti-H1N1 influenza virus antibody titer was determined using an indirect ELISA method.

[0071] The results showed that the control composition had a titer of 1:32000 before nebulization, which decreased to 1:25600 after nebulization, with a retention rate of approximately 80%. The experimental group had a titer of 1:32000 before nebulization, which decreased to 1:29600 after nebulization, with a retention rate of approximately 92.5%. The experimental group showed a titer retention rate increase of approximately 12.5 percentage points compared to the control group. This indicates that the addition of natural components significantly protected the antibody activity during nebulization.

[0072] Respiratory mucosal retention time measurement:

[0073] Fluorescent labeling experiments were conducted using an isolated porcine tracheal mucosa model. Fresh porcine trachea was longitudinally cut to expose the mucosal surface. FITC-labeled multi-target complex yolk globulin (F / P ratio = 3-5) was prepared into two formulations: one containing sodium hyaluronate (Example 1) and the other a control without sodium hyaluronate. 0.1 mL of each was added to the mucosal surface. Initial fluorescence intensity was measured using a fluorescence microscope, and then every 10 minutes until the fluorescence intensity decreased to 50% of its initial value. As an indicator of mucosal retention. Each group was divided into 3 parallel groups, and the experiment was repeated 3 times.

[0074] The results showed that the half-life time of the control group was approximately 42 minutes, while that of the experimental group was approximately 55 minutes, an increase of about 31%. This indicates that sodium hyaluronate can effectively prolong the retention time of antibodies in the respiratory mucosa, thereby enhancing the local effect.

[0075] Validation of the synergistic effect of functional gain components:

[0076] To verify the effects of different natural components on antibody function, multiple additive groups were established. Equal volumes of multi-target complex egg yolk globulin solution were taken, and equal volumes of Polygonatum odoratum extract dilution, sodium hyaluronate dilution, steviol glycoside dilution, and pear juice dilution were added, respectively. An antibody solution without any additives served as a control. Each sample was mixed with an equal volume of influenza H1N1 virus solution for virus neutralization experiments. A 2-fold serial dilution method was used, with 8 replicate wells per dilution. The half-maximal neutralizing titer (NT50) was calculated using the Reed-Muench method, with cytopathic effect (CPE) as the criterion.

[0077] The results are as follows: The NT50 of the control group was 1:6400; after adding the dilution of Polygonatum odoratum extract, the NT50 increased to 1:7680, an increase of approximately 20%; after adding the dilution of sodium hyaluronate, the NT50 increased to 1:7040, an increase of approximately 10%; after adding the dilution of steviol glycosides, the NT50 was 1:6592, a change of less than 3%, which was not statistically significant; after adding the dilution of pear juice, the NT50 was 1:6528, also without significant change. These results confirm that Polygonatum odoratum extract and sodium hyaluronate have clear immune synergistic or antibody protective functions, while components such as steviol glycosides and pear juice, which only provide taste or physical soothing effects, do not possess functional gains.

[0078] To further illustrate the influence of the content of each component in the composition of this invention on the overall effect, four other compositions with different parameters were formulated and compared with the above-mentioned top-level T0 formulation. All formulations were prepared using the same method, with only the concentration of the multi-target complex egg yolk globulin powder and the amount of natural ingredients added being varied. The specific parameters of each formulation are shown in Table 1.

[0079] Table 1. Performance comparison of formulations with different parameters:

[0080] Formula A 0.5 0.10 0.05 93.1 32.5 Formula B 1.0 0.10 0.05 92.5 31.2 Formula C 2.0 0.10 0.05 90.8 29.6 Formula D 1.0 0.05 0.025 91.2 18.3 Formula E 1.0 0.20 0.10 93.7 39.4

[0081] As shown in Table 1, within the antibody concentration range of 0.5% to 2.0%, the titer retention rate after nebulization was consistently above 90%. When the antibody concentration increased to 2.0%, the retention rate decreased slightly, possibly due to increased intermolecular interactions at higher concentrations, but it remained higher than the 80% of the control group. Regarding the amount of natural ingredients added, increasing the amount of Polygonatum odoratum extract and sodium hyaluronate improved both the nebulization protective effect and the mucosal retention time. However, considering cost and the tolerability of sensitive individuals, 0.10 mL and 0.05 mL were chosen as the optimal balance points in the examples. After halving the amount of natural ingredients in Formulation D, the mucosal half-life extension rate decreased significantly, indicating a positive correlation between the protective effect of sodium hyaluronate and its addition amount.

[0082] Based on the comprehensive verification results, the formulation and preparation method described in this embodiment can achieve high activity retention after antibody atomization, long-term retention in the respiratory mucosa, and synergistic enhancement of immunity by natural ingredients, and is feasible for actual production and clinical application.

[0083] like Figure 2 As shown, the nebulization delivery process of the composition of the present invention is illustrated as follows: The nebulizer converts the liquid preparation into aerosol particles with a particle size of 1-5 micrometers, which carry antibodies and natural ingredients and settle on the surface of the respiratory mucosa. The antibodies neutralize pathogens immediately, and the natural ingredients begin to exert their mucosal repair and immune regulation effects.

[0084] like Figure 3As shown, the dual-pathway synergistic mechanism of the present invention can be summarized as follows: the passive neutralizing layer blocks invasion by binding antibodies to pathogen surface antigens; the active gain layer regulates gut-lung axis immunity through Polygonatum odoratum polysaccharide, repairs physical barriers through sodium hyaluronate, and inhibits inflammatory factors through plant extracts, and the three work together to prolong the protective effect.

[0085] The multi-target complex egg yolk globulin nebulized inhalation composition for combating respiratory infections provided in this embodiment is prepared by co-dissolving a broad-spectrum antibody prepared through combined immunization with food-grade natural immune-enhancing components in an isotonic aqueous solution. The mixture is then formulated using a low-temperature, mild preparation process and microporous filtration. The resulting product exhibits stable physicochemical properties, with an antibody titer retention rate of no less than 90% after nebulization, and a mucosal retention time that is more than 30% longer than traditional formulations. Animal-level virus neutralization experiments show that the addition of Polygonatum odoratum extract can increase the neutralization titer by approximately 15%. All the above data are from specific tests conducted in this embodiment, and the detection methods used are conventional methods in the art; the experimental results are reproducible.

[0086] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention in any way. Although the present invention has been disclosed above with reference to preferred embodiments, it is not intended to limit the present invention. Any person skilled in the art can make some modifications or alterations to the above-disclosed technical content to create equivalent embodiments without departing from the scope of the present invention. Any simple modifications, equivalent changes and alterations made to the above embodiments based on the technical essence of the present invention without departing from the scope of the present invention shall still fall within the scope of the present invention.

Claims

1. A multi-target complex yolk globulin for combating respiratory infections, characterized in that, This multi-target complex egg yolk globulin is a broad-spectrum anti-respiratory pathogen multi-target chicken-derived inducible-natural complex antibody nebulization inhalation composition containing food-grade natural immune-enhancing components. The composition comprises the following components: A multi-target complex chicken-derived inducible-natural complex antibody with a content of 0.5%-2.5% w / v, wherein the multi-target complex chicken-derived inducible-natural complex antibody contains specific antibodies against viral antigens and bacterial antigens; At least one food-grade natural immune-enhancing ingredient, with a content of 0.01%-1.0% v / v, wherein the food-grade natural immune-enhancing ingredient is selected from one or more of plant-derived immunomodulatory polysaccharides, food-grade mucosal repair factors, and food-grade antioxidant and anti-inflammatory plant extracts; Sodium chloride is used as an isotonic regulator at a concentration of 0.9% w / v; The remainder is purified water; The multi-target complex chicken-derived inducible-natural complex antibody and the food-grade natural immune-enhancing ingredient are co-soluble in the same aqueous matrix and act synchronously on the respiratory mucosa after being atomized by ultrasound or micro-mesh; and the food-grade natural immune-enhancing ingredient does not include steviosides, fruit juices, or menthol substances that only provide taste or physical relief and have no positive antibody effect.

2. The multi-target complex egg yolk globulin for resisting respiratory tract infections according to claim 1, characterized in that, The plant-derived immunomodulatory polysaccharides include Polygonatum odoratum extract, which is pre-prepared as a 50-200 times diluted standard solution, and the amount added per 5 mL of composition is 0.01-0.15 mL; The food-grade mucosal repair factor includes sodium hyaluronate, which is pre-prepared as a 150-300 times diluted standard solution, and the amount added per 5 mL of composition is 0.01-0.08 mL.

3. The multi-target complex egg yolk globulin for resisting respiratory tract infections according to claim 1, characterized in that, The food-grade antioxidant and anti-inflammatory plant extract is selected from one or more of the following: perilla extract, lily extract, monk fruit extract, and honeysuckle extract. It is prepared in advance as a 100-200 times diluted standard solution, and the amount added per 5 mL of composition is 0.02-0.10 mL. The composition also contains one or more auxiliary stabilizing ingredients selected from erythritol solution, food-grade glycerin, and food-grade lactic acid.

4. The multi-target complex egg yolk globulin for resisting respiratory tract infections according to claim 1, characterized in that, In a single-preparation composition, the number of functional ingredients in the food-grade natural immune-enhancing ingredient does not exceed two.

5. The multi-target complex egg yolk globulin for resisting respiratory tract infections according to claim 1, characterized in that, The viral antigen group includes at least three of the following: influenza virus antigen, novel coronavirus antigen, respiratory syncytial virus antigen, adenovirus antigen, and rhinovirus antigen. The bacterial antigen group includes at least two of the following: Streptococcus pneumoniae antigen, Haemophilus influenzae antigen, group A type B hemolytic streptococcal antigen, and Staphylococcus aureus antigen.

6. The multi-target complex egg yolk globulin for resisting respiratory tract infections according to claim 1, characterized in that, The viral antigen set includes conserved viral protein region antigens, which are selected from one or more of the following: influenza virus HA2 fusion peptide, novel coronavirus S2 conserved region, and respiratory syncytial virus F protein conserved epitope.

7. The multi-target complex egg yolk globulin for resisting respiratory tract infections according to claim 1, characterized in that, The composition has a pH of 5.5-7.0, an osmotic pressure of 280-320 mOsm / kg, and no visible particulate matter after filtration through a 0.22 μm microporous membrane.

8. A method for preparing a multi-target complex egg yolk globulin for combating respiratory infections, applicable to the multi-target complex egg yolk globulin for combating respiratory infections as described in any one of claims 1 to 7, characterized in that, The method includes the following steps: S1, Dissolve sodium chloride in purified water to prepare a 0.9% isotonic aqueous solution; S2, add one or two pre-diluted food-grade natural immune-enhancing ingredients to the solution of S1 in sequence, and stir well; S3, add multi-target complex chicken-derived inducible-natural complex antibody powder to the mixture of S2, and gently stir at a low temperature of 2-8℃ until completely dissolved; S4, add purified water to the target volume, filter through a 0.22μm microporous membrane, fill and seal to obtain the product.

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