Compound exosome temperature-sensitive gel nasal delivery preparation and application
By combining exosomes and poloxamer thermosensitive gel, the problems of short drug retention time and high irritation to the nasal mucosa in nasal drug delivery formulations are solved, achieving long-term retention and synergistic therapeutic effects, thus improving the treatment efficacy of nasal diseases.
Patent Information
- Application Number
- CN202511580812.2
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-10-31
- Publication Date
- 2026-01-09
AI Technical Summary
Existing nasal delivery formulations have problems such as short drug retention time, easy clearance by cilia, and high irritation to the nasal mucosa, resulting in limited duration of efficacy and poor patient compliance.
The formula uses a combination of exosomes, poloxamer thermosensitive gel, and functional ingredients. The exosomes are derived from human mesenchymal stem cells and are engineered to form a gel layer that adheres to the nasal cavity for a long time. The functional ingredients, such as menthol and borneol, play a synergistic therapeutic role.
It improves drug stability and retention time, enhances the therapeutic effect on nasal diseases, solves the problems of drug stability and short retention time, and has important clinical application value.
Abstract
Description
Technical Field
[0001] This invention belongs to the field of biomedical technology, specifically a compound exosome thermosensitive gel nasal delivery formulation and its application. Background Technology
[0002] Nasal administration, as a non-invasive method of drug delivery, has many advantages. The nasal mucosa has a rich blood supply, allowing for rapid drug absorption. It can bypass the first-pass effect of the liver and directly enter the systemic circulation, improving the bioavailability of drugs. At the same time, the nasal cavity is directly connected to the central nervous system through the olfactory nerve and the trigeminal nerve, providing a unique drug delivery route for the treatment of some neurological diseases.
[0003] Currently, many drugs are used clinically via nasal administration, such as nasal corticosteroid sprays for treating allergic rhinitis. However, existing nasal administration formulations still have some limitations, such as short drug retention time, easy clearance by nasal cilia, resulting in limited duration of efficacy; some drugs are also highly irritating to the nasal mucosa, affecting patient compliance. Therefore, there is an urgent need to develop a new type of nasal administration compound formulation to solve problems such as drug stability, retention time, and limited therapeutic function. Summary of the Invention
[0004] To address the shortcomings of existing technologies, this invention provides a compound exosome thermosensitive gel nasal delivery formulation and its application, which has the advantages of drug stability, retention time, and diverse therapeutic functions.
[0005] To achieve the above objectives, the present invention provides the following technical solution: a compound exosome thermosensitive gel nasal delivery formulation, comprising the following components: exosomes, poloxamer thermosensitive gel, and functional ingredients.
[0006] As a preferred embodiment of the present invention, the functional ingredient is selected from one or more of menthol, borneol, and ginkgo leaf extract.
[0007] As a preferred embodiment of the present invention, the exosomes are derived from human mesenchymal stem cells, which have multi-directional differentiation potential and immunomodulatory function, and the exosomes derived from them have high biological activity and good safety.
[0008] As a preferred embodiment of the present invention, the exosomes need to be processed by an engineering method, which includes pretreatment of human mesenchymal stem cells: stimulation with specific cytokines to increase the secretion of exosomes and the content of specific bioactive molecules; and surface modification of exosomes: linking targeting molecules to make them act more specifically on nasal lesions.
[0009] As a preferred embodiment of the present invention, the method for preparing the nasal delivery formulation is as follows: Step 1: Exosomes were extracted from the culture supernatant of human mesenchymal stem cells using ultracentrifugation. First, human mesenchymal stem cells were cultured in serum-free medium, and the culture supernatant was collected. Then, low-speed centrifugation was performed to remove cell debris, and high-speed centrifugation was performed to precipitate the exosomes. Finally, the exosome precipitate was washed with phosphate buffer to obtain purified exosomes. Step Two: Poloxamer 407 and poloxamer 188 were dissolved in distilled water in a certain proportion and dissolved overnight in a refrigerator at 4°C to obtain a homogeneous poloxamer thermosensitive gel solution. The phase transition temperature of the poloxamer thermosensitive gel was determined by rheological testing. When the concentration of poloxamer 407 was 18%-22% and the concentration of poloxamer 188 was 2%-5%, the phase transition temperature of the gel was between 32-34°C, which is close to the human body temperature. Step 3: Dissolve menthol, borneol or ginkgo leaf extract in an organic solvent as needed, and then slowly add it to the poloxamer thermosensitive gel solution, stirring until homogeneous. Step Four: The purified exosome suspension was slowly added to the poloxamer thermosensitive gel solution containing the functional ingredient, and gently mixed to avoid exosome rupture, thus obtaining the compound exosome thermosensitive gel nasal administration formulation. The prepared compound exosome thermosensitive gel nasal administration formulation was stored in a refrigerator at 4°C for later use.
[0010] As a preferred embodiment of the present invention, the particle size distribution of the purified exosomes described in step one is detected by nanoparticle tracking analysis.
[0011] As a preferred embodiment of the present invention, the nasal delivery formulation is verified through animal experiments after preparation. The specific verification method is as follows: Experimental animals: Sixty SPF-grade SD rats, weighing 200-250g, were randomly divided into 6 groups: normal control group, model control group, compound group containing menthol, compound group containing borneol and ginkgo leaf extract, exosome group, and functional component group. Establishment of an allergic rhinitis model: A rat allergic rhinitis model was established using the ovalbumin sensitization method. Rats were sensitized by intraperitoneal injection of ovalbumin solution once a week for a total of 3 injections. Then, 1% ovalbumin solution was used for nasal provocation once a day for 10 consecutive days. Treatment: Rats in the normal control group and the model control group were given equal amounts of physiological saline nasal drops; the compound group containing menthol and the compound group containing borneol and ginkgo biloba extract were given the corresponding compound exosome thermosensitive gel composition nasal drops; the exosome group was given exosome suspension nasal drops; and the functional ingredient group was given solution containing the corresponding functional ingredient nasal drops. The administration was twice a day for 14 consecutive days. Observation indicators: Observe the nasal symptoms of rats: sneezing, runny nose, and scratching the nose, and then score them. At the same time, collect nasal secretions from rats and detect the levels of inflammatory factors (IL-4, IL-5, IL-13).
[0012] As a preferred embodiment of the present invention, the ovalbumin solution is a mixed solution containing 1 mg ovalbumin and 100 mg aluminum hydroxide.
[0013] As a preferred embodiment of the present invention, the above-mentioned compound exosome thermosensitive gel nasal delivery preparation is used in the treatment and prevention of nasal diseases, including but not limited to allergic rhinitis, chronic rhinitis, and sinusitis.
[0014] Compared with the prior art, the beneficial effects of the present invention are as follows: This invention combines exosomes, poloxamer thermosensitive gel, and functional ingredients. The exosomes are encapsulated in the poloxamer thermosensitive gel, preventing the influence of the external environment on their biological activity and improving the stability of the exosomes. The poloxamer thermosensitive gel forms a gel layer in the nasal cavity, allowing it to adhere to the nasal mucosa for a prolonged period, extending the drug's retention time. The functional ingredients and exosomes exert a synergistic therapeutic effect, improving the treatment efficacy for nasal diseases. Compared with existing nasal delivery formulations, this invention's compound exosome thermosensitive gel nasal delivery formulation can simultaneously solve problems related to drug stability, retention time, and limited therapeutic function, and is expected to provide a new and effective formulation for the treatment of nasal diseases, possessing significant clinical application value and socio-economic benefits. Detailed Implementation
[0015] Based on the embodiments of this invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of this invention.
[0016] Example 1 A compound exosome thermosensitive gel nasal delivery formulation comprises the following components: exosomes, poloxamer thermosensitive gel, and functional ingredients.
[0017] The combination of exosomes, poloxamer thermosensitive gel, and functional ingredients fully leverages the advantages of each component: exosomes provide bioactive therapeutic effects; poloxamer thermosensitive gel is a commonly used thermosensitive gel material with good biocompatibility and reversible sol-gel phase transition characteristics. Encapsulating drugs in poloxamer thermosensitive gel can improve drug stability and retention time at the administration site. The application of poloxamer thermosensitive gel enables sustained drug release and prolonged retention in the nasal cavity; the functional ingredients play a role in adjuvant therapy, improving nasal ventilation, and reducing inflammation.
[0018] The functional ingredient is selected from menthol.
[0019] Menthol has a cooling effect, which can improve nasal ventilation and reduce congestion and edema of the nasal mucosa.
[0020] Among them, exosomes are derived from human mesenchymal stem cells, which have multi-directional differentiation potential and immune regulatory functions. The exosomes derived from them have high biological activity and good safety.
[0021] Exosomes are extracellular vesicles with a diameter of 30-150 nm, secreted by various cells, and carry a wealth of bioactive molecules, such as proteins, nucleic acids, and lipids. Studies have shown that exosomes have a variety of biological functions, including immunomodulation, tissue repair, and anti-inflammation. In nasal cavity-related diseases, exosomes can exert therapeutic effects through mechanisms such as regulating immune responses and promoting mucosal repair.
[0022] Among them, exosomes need to be processed by engineering methods, which include pretreatment of human mesenchymal stem cells: stimulation with specific cytokines to increase the secretion of exosomes and the content of specific bioactive molecules; and surface modification of exosomes: linking targeting molecules to make them act more specifically on nasal lesions.
[0023] Exosomes derived from human mesenchymal stem cells can alleviate nasal inflammation and promote the repair of damaged nasal mucosa. However, the use of exosomes alone has problems such as poor stability and short retention time in the nasal cavity, which limits their clinical application.
[0024] The preparation method of the nasal delivery formulation is as follows: Step 1: Exosomes were extracted from the culture supernatant of human mesenchymal stem cells using ultracentrifugation. First, human mesenchymal stem cells were cultured in serum-free medium, and the culture supernatant was collected. Then, low-speed centrifugation was performed to remove cell debris, and high-speed centrifugation was performed to precipitate the exosomes. Finally, the exosome precipitate was washed with phosphate buffer to obtain purified exosomes. Step Two: 20g of poloxamer 407 and 3g of poloxamer 188 were dissolved in 100ml of distilled water and dissolved overnight in a refrigerator at 4℃ to obtain a homogeneous poloxamer thermosensitive gel solution. The phase transition temperature of the poloxamer thermosensitive gel was determined to be 33℃ by rheological testing. Step 3: Dissolve 0.5g of menthol in 5ml of ethanol, then slowly add it to the poloxamer thermosensitive gel solution and stir until homogeneous. Step Four: 5 ml of purified exosome suspension was slowly added to the poloxamer thermosensitive gel solution containing the functional ingredient and gently mixed to obtain the compound exosome thermosensitive gel nasal administration formulation. The prepared compound exosome thermosensitive gel nasal administration formulation was stored in a refrigerator at 4°C for later use.
[0025] Poloxamer 407 and Poloxamer 188 are both nonionic block copolymer surfactants, prepared by chemical synthesis of propylene oxide, propylene glycol, and ethylene oxide. Poloxamer 407 is a fluid at low temperatures, but solidifies into a reversible physical gel upon heating, making it suitable for mucosal drug delivery systems. Poloxamer 188 can increase the solubility and stability of poorly soluble drugs and control the drug release rate.
[0026] In step one, the particle size distribution of the purified exosomes is detected by nanoparticle tracking analysis.
[0027] Nanoparticle tracking analysis is a technique that uses light scattering and Brownian motion properties to perform real-time dynamic characterization of nanoparticles in liquid suspensions. It tracks the Brownian motion trajectory of individual particles and calculates the hydrodynamic radius of the particles using the Stokes-Einstein equation. It also provides key parameters such as particle size distribution, concentration, and scattered light intensity. By detecting the particle size distribution of exosomes through nanoparticle tracking analysis, the results show that the exosome particle size is mainly concentrated between 30-150 nm, and the purity reaches more than 90%.
[0028] The nasal delivery formulation was prepared and then validated through animal experiments. The specific validation method was as follows: Experimental animals: Sixty SPF-grade SD rats, weighing 200-250g, were randomly divided into 6 groups: normal control group, model control group, compound group containing menthol, compound group containing borneol and ginkgo leaf extract, exosome group, and functional component group. Establishment of an allergic rhinitis model: A rat allergic rhinitis model was established using the ovalbumin sensitization method. Rats were sensitized by intraperitoneal injection of ovalbumin solution once a week for a total of 3 injections. Then, 1% ovalbumin solution was used for nasal provocation once a day for 10 consecutive days. Treatment: Rats in the normal control group and the model control group were given equal amounts of physiological saline nasal drops; the compound group containing menthol and the compound group containing borneol and ginkgo biloba extract were given the corresponding compound exosome thermosensitive gel composition nasal drops; the exosome group was given exosome suspension nasal drops; and the functional ingredient group was given solution containing the corresponding functional ingredient nasal drops. The administration was twice a day for 14 consecutive days. Observation indicators: Observe the nasal symptoms of rats: sneezing, runny nose, and scratching the nose, and then score them. At the same time, collect nasal secretions from rats and detect the levels of inflammatory factors (IL-4, IL-5, IL-13).
[0029] Experimental results: The results showed that, compared with the model control group, the nasal symptom scores of rats in the compound group containing menthol and the compound group containing borneol and ginkgo leaf extract were significantly reduced (P<0.05), and the levels of inflammatory factors in nasal secretions were also significantly reduced (P<0.05). Although the exosome group and the functional component group also had certain therapeutic effects, they were not as significant as those in the compound group. This indicates that the compound exosome thermosensitive gel nasal delivery formulation of the present invention has a better therapeutic effect on allergic rhinitis.
[0030] The ovalbumin solution is a mixed solution containing 1 mg of ovalbumin and 100 mg of aluminum hydroxide.
[0031] The ovalbumin sensitization method for establishing a rat model of allergic rhinitis is a classic method for studying allergic rhinitis by simulating the human allergic reaction mechanism. Its core lies in using ovalbumin as an antigen to induce an immune response and symptoms similar to human allergic rhinitis in rats through specific pathways and procedures. The principle is as follows: both allergic rhinitis and asthma are IgE-mediated type I hypersensitivity diseases. When ovalbumin enters the rat's body as an antigen, its soluble antigenic components stimulate the body to produce specific IgE antibodies. These antibodies attach to the surface of mast cells and basophils, sensitizing the rat. When the rat is exposed to the same antigen again, an antigen-antibody reaction occurs between the antigen and the IgE on the surface of the sensitized cells, leading to cell degranulation and the release of inflammatory mediators such as histamine and eosinophil chemotactic factors. These mediators act on the nasal mucosa, causing vasodilation, increased permeability, increased mucus secretion, and eosinophil infiltration, thereby producing the symptoms of allergic rhinitis.
[0032] Among them, the above-mentioned compound exosome thermosensitive gel nasal delivery preparation is used in the treatment and prevention of nasal diseases, including but not limited to allergic rhinitis, chronic rhinitis, and sinusitis.
[0033] Nasal administration allows medication to act directly on the lesion site, exerting anti-inflammatory, immunomodulatory, and mucosal repair-promoting effects, thereby improving treatment efficacy and reducing systemic adverse reactions.
[0034] Example 2 A compound exosome thermosensitive gel nasal delivery formulation comprises the following components: exosomes, poloxamer thermosensitive gel, and functional ingredients.
[0035] The combination of exosomes, poloxamer thermosensitive gel, and functional ingredients fully leverages the advantages of each component: exosomes provide bioactive therapeutic effects; poloxamer thermosensitive gel is a commonly used thermosensitive gel material with good biocompatibility and reversible sol-gel phase transition characteristics. Encapsulating drugs in poloxamer thermosensitive gel can improve drug stability and retention time at the administration site. The application of poloxamer thermosensitive gel enables sustained drug release and prolonged retention in the nasal cavity; the functional ingredients play a role in adjuvant therapy, improving nasal ventilation, and reducing inflammation.
[0036] The functional ingredients are selected from borneol and ginkgo leaf extract.
[0037] Borneol has the effects of opening the orifices and refreshing the mind, clearing heat and relieving pain, and can promote the absorption of drugs in the nasal cavity; Ginkgo biloba extract contains active ingredients such as flavonoids and terpene lactones, which have anti-inflammatory, antioxidant and microcirculation-improving effects.
[0038] Among them, exosomes are derived from human mesenchymal stem cells, which have multi-directional differentiation potential and immune regulatory functions. The exosomes derived from them have high biological activity and good safety.
[0039] Exosomes are extracellular vesicles with a diameter of 30-150 nm, secreted by various cells, and carry a wealth of bioactive molecules, such as proteins, nucleic acids, and lipids. Studies have shown that exosomes have a variety of biological functions, including immunomodulation, tissue repair, and anti-inflammation. In nasal cavity-related diseases, exosomes can exert therapeutic effects through mechanisms such as regulating immune responses and promoting mucosal repair.
[0040] Among them, exosomes need to be processed by engineering methods, which include pretreatment of human mesenchymal stem cells: stimulation with specific cytokines to increase the secretion of exosomes and the content of specific bioactive molecules; and surface modification of exosomes: linking targeting molecules to make them act more specifically on nasal lesions.
[0041] Exosomes derived from human mesenchymal stem cells can alleviate nasal inflammation and promote the repair of damaged nasal mucosa. However, the use of exosomes alone has problems such as poor stability and short retention time in the nasal cavity, which limits their clinical application.
[0042] The preparation method of the nasal delivery formulation is as follows: Step 1: Exosomes were extracted from the culture supernatant of human mesenchymal stem cells using ultracentrifugation. First, human mesenchymal stem cells were cultured in serum-free medium, and the culture supernatant was collected. Then, low-speed centrifugation was performed to remove cell debris, and high-speed centrifugation was performed to precipitate the exosomes. Finally, the exosome precipitate was washed with phosphate buffer to obtain purified exosomes. Step Two: 19g of poloxamer 407 and 4g of poloxamer 188 were dissolved in 100ml of distilled water and dissolved overnight in a refrigerator at 4℃ to obtain a homogeneous poloxamer thermosensitive gel solution. The phase transition temperature of the poloxamer thermosensitive gel was determined to be 32.5℃ by rheological testing. Step 3: Weigh 0.3g of borneol and 1g of ginkgo leaf extract, dissolve them separately in an appropriate amount of ethanol, mix them evenly, and slowly add them to the poloxamer thermosensitive gel solution, stirring evenly. Step Four: 5 ml of purified exosome suspension was slowly added to the poloxamer thermosensitive gel solution containing the functional ingredient and gently mixed to obtain the compound exosome thermosensitive gel nasal administration formulation. The prepared compound exosome thermosensitive gel nasal administration formulation was stored in a refrigerator at 4°C for later use.
[0043] Poloxamer 407 and Poloxamer 188 are both nonionic block copolymer surfactants, prepared by chemical synthesis of propylene oxide, propylene glycol, and ethylene oxide. Poloxamer 407 is a fluid at low temperatures, but solidifies into a reversible physical gel upon heating, making it suitable for mucosal drug delivery systems. Poloxamer 188 can increase the solubility and stability of poorly soluble drugs and control the drug release rate.
[0044] In step one, the particle size distribution of the purified exosomes is detected by nanoparticle tracking analysis.
[0045] Nanoparticle tracking analysis is a technique that uses light scattering and Brownian motion properties to perform real-time dynamic characterization of nanoparticles in liquid suspensions. It tracks the Brownian motion trajectory of individual particles and calculates the hydrodynamic radius of the particles using the Stokes-Einstein equation. It also provides key parameters such as particle size distribution, concentration, and scattered light intensity. By detecting the particle size distribution of exosomes through nanoparticle tracking analysis, the results show that the exosome particle size is mainly concentrated between 30-150 nm, and the purity reaches more than 90%.
[0046] The nasal delivery formulation was prepared and then validated through animal experiments. The specific validation method was as follows: Experimental animals: Sixty SPF-grade SD rats, weighing 200-250g, were randomly divided into 6 groups: normal control group, model control group, compound group containing menthol, compound group containing borneol and ginkgo leaf extract, exosome group, and functional component group. Establishment of an allergic rhinitis model: A rat allergic rhinitis model was established using the ovalbumin sensitization method. Rats were sensitized by intraperitoneal injection of ovalbumin solution once a week for a total of 3 injections. Then, 1% ovalbumin solution was used for nasal provocation once a day for 10 consecutive days. Treatment: Rats in the normal control group and the model control group were given equal amounts of physiological saline nasal drops; the compound group containing menthol and the compound group containing borneol and ginkgo biloba extract were given the corresponding compound exosome thermosensitive gel composition nasal drops; the exosome group was given exosome suspension nasal drops; and the functional ingredient group was given solution containing the corresponding functional ingredient nasal drops. The administration was twice a day for 14 consecutive days. Observation indicators: Observe the nasal symptoms of rats: sneezing, runny nose, and scratching the nose, and then score them. At the same time, collect nasal secretions from rats and detect the levels of inflammatory factors (IL-4, IL-5, IL-13).
[0047] Experimental results: The results showed that, compared with the model control group, the nasal symptom scores of rats in the compound group containing menthol and the compound group containing borneol and ginkgo leaf extract were significantly reduced (P<0.05), and the levels of inflammatory factors in nasal secretions were also significantly reduced (P<0.05). Although the exosome group and the functional component group also had certain therapeutic effects, they were not as significant as those in the compound group. This indicates that the compound exosome thermosensitive gel nasal delivery formulation of the present invention has a better therapeutic effect on allergic rhinitis.
[0048] The ovalbumin solution is a mixed solution containing 1 mg of ovalbumin and 100 mg of aluminum hydroxide.
[0049] The ovalbumin sensitization method for establishing a rat model of allergic rhinitis is a classic method for studying allergic rhinitis by simulating the human allergic reaction mechanism. Its core lies in using ovalbumin as an antigen to induce an immune response and symptoms similar to human allergic rhinitis in rats through specific pathways and procedures. The principle is as follows: both allergic rhinitis and asthma are IgE-mediated type I hypersensitivity diseases. When ovalbumin enters the rat's body as an antigen, its soluble antigenic components stimulate the body to produce specific IgE antibodies. These antibodies attach to the surface of mast cells and basophils, sensitizing the rat. When the rat is exposed to the same antigen again, an antigen-antibody reaction occurs between the antigen and the IgE on the surface of the sensitized cells, leading to cell degranulation and the release of inflammatory mediators such as histamine and eosinophil chemotactic factors. These mediators act on the nasal mucosa, causing vasodilation, increased permeability, increased mucus secretion, and eosinophil infiltration, thereby producing the symptoms of allergic rhinitis.
[0050] Among them, the above-mentioned compound exosome thermosensitive gel nasal delivery preparation is used in the treatment and prevention of nasal diseases, including but not limited to allergic rhinitis, chronic rhinitis, and sinusitis.
[0051] Nasal administration allows medication to act directly on the lesion site, exerting anti-inflammatory, immunomodulatory, and mucosal repair-promoting effects, thereby improving treatment efficacy and reducing systemic adverse reactions.
[0052] It should be noted that, in this document, relational terms such as "first" and "second" are used only to distinguish one entity or operation from another, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Furthermore, the terms "comprising," "including," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such process, method, article, or apparatus.
[0053] Although embodiments of the invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the appended claims and their equivalents.
Claims
1. A compound exosome thermosensitive gel nasal delivery formulation, characterized in that, It includes the following components: exosomes, poloxamer thermosensitive gel, and functional ingredients.
2. The compound exosome thermosensitive gel nasal delivery formulation according to claim 1, characterized in that: The functional ingredients are selected from one or more of menthol, borneol, and ginkgo leaf extract.
3. The compound exosome thermosensitive gel nasal delivery formulation according to claim 1, characterized in that: The exosomes are derived from human mesenchymal stem cells, which have multi-directional differentiation potential and immunomodulatory functions. The exosomes derived from these stem cells have high biological activity and good safety.
4. The compound exosome thermosensitive gel nasal delivery formulation according to claim 1, characterized in that: The exosomes need to be processed by an engineering method, which includes pretreatment of human mesenchymal stem cells: stimulation with specific cytokines to increase the secretion of exosomes and the content of specific bioactive molecules; and surface modification of exosomes: linking targeting molecules to make them act more specifically on nasal lesions.
5. The compound exosome thermosensitive gel nasal delivery formulation according to claim 1, characterized in that: The method for preparing the nasal delivery formulation is as follows: Step 1: Exosomes were extracted from the culture supernatant of human mesenchymal stem cells using ultracentrifugation. First, human mesenchymal stem cells were cultured in serum-free medium, and the culture supernatant was collected. Then, low-speed centrifugation was performed to remove cell debris, and high-speed centrifugation was performed to precipitate the exosomes. Finally, the exosome precipitate was washed with phosphate buffer to obtain purified exosomes. Step Two: Poloxamer 407 and poloxamer 188 were dissolved in distilled water in a certain proportion and dissolved overnight in a refrigerator at 4°C to obtain a homogeneous poloxamer thermosensitive gel solution. The phase transition temperature of the poloxamer thermosensitive gel was determined by rheological testing. When the concentration of poloxamer 407 was 18%-22% and the concentration of poloxamer 188 was 2%-5%, the phase transition temperature of the gel was between 32-34°C, which is close to the human body temperature. Step 3: Dissolve menthol, borneol or ginkgo leaf extract in an organic solvent as needed, and then slowly add it to the poloxamer thermosensitive gel solution, stirring until homogeneous. Step Four: The purified exosome suspension was slowly added to the poloxamer thermosensitive gel solution containing the functional ingredient, and gently mixed to avoid exosome rupture, thus obtaining the compound exosome thermosensitive gel nasal administration formulation. The prepared compound exosome thermosensitive gel nasal administration formulation was stored in a refrigerator at 4°C for later use.
6. The compound exosome thermosensitive gel nasal delivery formulation according to claim 5, characterized in that: The particle size distribution of the purified exosomes described in step one was detected by nanoparticle tracking analysis.
7. The compound exosome thermosensitive gel nasal delivery formulation according to claim 5, characterized in that: After the nasal delivery formulation was prepared, it was validated through animal experiments. The specific validation method was as follows: Experimental animals: Sixty SPF-grade SD rats, weighing 200-250g, were randomly divided into 6 groups: normal control group, model control group, compound group containing menthol, compound group containing borneol and ginkgo leaf extract, exosome group, and functional component group. Establishment of an allergic rhinitis model: A rat allergic rhinitis model was established using the ovalbumin sensitization method. Rats were sensitized by intraperitoneal injection of ovalbumin solution once a week for a total of 3 injections. Then, 1% ovalbumin solution was used for nasal provocation once a day for 10 consecutive days. Treatment: Rats in the normal control group and the model control group were given equal amounts of physiological saline nasal drops; the compound group containing menthol and the compound group containing borneol and ginkgo biloba extract were given the corresponding compound exosome thermosensitive gel composition nasal drops; the exosome group was given exosome suspension nasal drops; and the functional ingredient group was given solution containing the corresponding functional ingredient nasal drops. The administration was twice a day for 14 consecutive days. Observation indicators: Observe the nasal symptoms of rats: sneezing, runny nose, and scratching the nose, and then score them. At the same time, collect nasal secretions from rats and detect the levels of inflammatory factors (IL-4, IL-5, IL-13).
8. The compound exosome thermosensitive gel nasal delivery formulation according to claim 7, characterized in that: The ovalbumin solution is a mixed solution containing 1 mg of ovalbumin and 100 mg of aluminum hydroxide.
9. The application of a compound exosome thermosensitive gel nasal delivery formulation according to any one of claims 1-8 in the treatment and prevention of nasal diseases, characterized in that, Including but not limited to allergic rhinitis, chronic rhinitis, and sinusitis.