Exosome composition containing baicalein or derivative thereof, and use thereof in treating rhinitis

Abstract

An exosome composition containing baicalein or a derivative thereof, and a use thereof in treating rhinosinusitis. Baicalein or a derivative thereof in the exosome composition is partially or completely loaded in an exosome. The baicalein or the derivative thereof and the exosome can achieve a synergistic treatment effect, thereby achieving a good treatment effect on rhinosinusitis, especially on eosinophilic chronic rhinosinusitis.

Description

An exosome composition containing baicalein or its derivatives and its application in treating rhinitis

[0001] This application claims priority to a prior application, patent application number 202311227597.9, filed with the State Intellectual Property Office of China on September 22, 2023, entitled "An exosome composition containing baicalein or its derivatives and its use in treating rhinitis." The entire text of that prior application is incorporated herein by reference. Technical Field

[0002] The present invention relates to the field of medical technology, and in particular to an exosome composition comprising baicalein or a derivative thereof and an application of the exosome composition, especially an application in treating rhinitis. Background Art

[0003] The anatomical structures of the nasal cavity and sinuses are interconnected. Therefore, while a distinction is made clinically between rhinitis and sinusitis, they are often interrelated and often occur simultaneously. Following the release of the European Academy of Allergy and Clinical Immunology's evidence-based diagnosis and treatment guidelines for sinusitis and nasal polyps in 2007, rhinitis and sinusitis have been collectively referred to as rhinosinusitis (including nasal polyps) in clinical practice both domestically and internationally. This refers to inflammation of the nasal cavity and sinuses, manifested by two or more clinical features of nasal and sinus inflammation: nasal obstruction / congestion and / or rhinorrhea (anterior and posterior rhinorrhea), with or without facial tenderness / swelling, and with or without decreased or loss of smell; nasal polyps, mucopurulent discharge from the middle nasal meatus, and / or swelling or obstruction of the middle nasal meatus mucosa on nasal endoscopy; and changes in the ostiomeatal complex or sinus mucosa on nasal CT scan. Symptoms are categorized as acute or chronic based on the duration of symptoms. Acute rhinosinusitis refers to symptoms lasting less than 12 weeks with complete remission, while chronic rhinosinusitis refers to symptoms lasting more than 12 weeks with incomplete remission or possible exacerbation. The 2020 European Opinion on Sinusitis and Nasal Polyps classifies chronic sinusitis into type 1 (Th1), type 2 (Th2), and type 3 (Th17) based on different immune inflammation types. Type 2 is also commonly referred to as eosinophilic chronic sinusitis in clinical practice, while types 1 and 3 are also referred to as non-type 2, which are non-eosinophilic chronic sinusitis.

[0004] Although the current routine clinical treatment for chronic sinusitis is still the topical use of glucocorticoids as the first-line treatment, and no more precise symptomatic treatment is provided for different subtypes, the efficacy of these treatments varies greatly due to the different immune responses and inflammatory characteristics of chronic sinusitis subtypes, such as type 2 inflammation and non-type 2 inflammation, causing many patients with insensitive responses to receive treatments that are actually ineffective.

[0005] Baicalein is one of the most abundant flavonoids in Scutellaria baicalensis. Baicalin is a glycoside formed by the combination of baicalein and a molecule of glucuronic acid. Both are present in Scutellaria baicalensis. Baicalein and baicalin have similar pharmacological activities, including anti-inflammatory, antioxidant, and free radical scavenging properties, and both undergo extensive biotransformation in the body.

[0006] Similar to mesenchymal stem cells, induced pluripotent stem cells (iPS) possess robust differentiation and regeneration capabilities. However, iPS cells possess the property of unlimited proliferation, allowing them to differentiate into the various cell types required by the human body's organs and tissues. iPS cells are not directly derived from human organs and tissues, but rather stem cells derived through gene editing. Therefore, there are no ethical issues surrounding their origin. Furthermore, iPS cells are easier to obtain than mesenchymal stem cells. Therefore, iPS exosomes are easier to produce on a large scale and are more readily available than mesenchymal stem cell exosomes.

[0007] Exosomes are extracellular vesicles secreted by cells. They possess unique properties such as low immunogenicity, inherent stability, high delivery efficiency, and the ability to cross the blood-brain barrier. They also possess numerous advantages, including tissue repair and anti-inflammatory properties. Exosomes have also garnered extensive attention and research as drug nanocarriers.

[0008] Summary of the Invention

[0009] The present invention relates to an exosome composition containing baicalein or a derivative thereof and its use in treating rhinosinusitis, particularly eosinophilic chronic sinusitis. Using exosomes to encapsulate baicalein or a derivative thereof can improve the solubility of baicalein or a derivative thereof, prolong its circulation time in the body, retain its therapeutic activity, and enhance the permeability of baicalein or a derivative thereof in the nasal mucosa. The exosome composition containing baicalein or a derivative thereof is more effective in treating rhinosinusitis than the therapeutic effects of exosomes alone, baicalein or a derivative thereof alone, or the sequential or simultaneous administration of independent exosomes and independent baicalein or a derivative thereof.

[0010] A first aspect of the present invention is to provide an exosome composition comprising baicalein or a derivative thereof, wherein the exosome composition comprises a combination of baicalein or a derivative thereof and exosomes as an active ingredient. In one embodiment of the present invention, only a portion of the baicalein or a derivative thereof in the exosome composition is loaded into the exosomes. In another embodiment of the present invention, all of the baicalein or a derivative thereof in the exosome composition is loaded into the exosomes. In the present invention, "loaded into the exosomes" includes at least two forms: baicalein or a derivative thereof permeates into the exosomes, and baicalein or a derivative thereof is associated with the exosomes.

[0011] In some embodiments of the present invention, the loading concentration of baicalein or its derivatives is 2-500 μg / mL, for example, 2.1 μg / mL, 2.5 μg / mL, 3 μg / mL, 4 μg / mL, 5 μg / mL, 6 μg / mL, 7 μg / mL, 8 μg / mL, 9 μg / mL, 10 μg / mL, 20 μg / mL, 30 μg / mL, 40 μg / mL, 50 μg / mL, 60 μg / mL, 70 μg / mL, 80 μg / mL, 90 μg / mL, 100 μg / mL, mL, 110μg / mL, 120μg / mL, 130μg / mL, 140μg / mL, 150μg / mL, 160μg / mL, 170μg / mL, 180μg / mL, 190μg / mL, 200μg / mL, 210 μg / mL, 220μg / mL, 230μg / mL, 240μg / mL, 250μg / mL, 260μg / mL, 270μg / mL, 280μg / mL, 290μg / mL, 300μg / mL, 310μg / mL, 320 μg / mL, 330 μg / mL, 340 μg / mL, 350 μg / mL, 360 μg / mL, 370 μg / mL, 380 μg / mL, 390 μg / mL, 400 μg / mL, 410 μg / mL, 420 μg / mL, 430 μg / mL, 440 μg / mL, 450 μg / mL, 460 μg / mL, 470 μg / mL, 480 μg / mL, 490 μg / mL, 495 μg / mL, etc. In the present invention, the “loading concentration” refers to the concentration of baicalein or its derivatives loaded in the exosomes in the exosome composition.

[0012] In some embodiments of the present invention, in the exosome composition, the amount of baicalein or its derivatives loaded in the exosomes, calculated as baicalein, accounts for 5%-100% of the total amount of baicalein or its derivatives in the exosome composition, for example, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, etc.

[0013] The present invention also provides an exosome composition comprising baicalein or its derivatives for treating rhinosinusitis. The exosome composition comprising baicalein or its derivatives is as described above.

[0014] The second aspect of the present invention is to provide the use of the exosome composition containing baicalein or its derivatives in the preparation of a medicament for treating rhinosinusitis;

[0015] Also provided is the use of exosomes and baicalein or its derivatives in preparing a medicine for treating rhinosinusitis.

[0016] The third aspect of the present invention is to provide a method for preparing the exosome composition containing baicalein or its derivatives.

[0017] In some embodiments of the present invention, the exosome composition comprising baicalein or its derivatives is prepared by incubation, sonication, freeze-thaw, extrusion or saponin treatment.

[0018] In one embodiment of the present invention, the incubation method comprises: preparing a solution of baicalein or a derivative thereof at an appropriate concentration and co-incubating it with exosomes. In one embodiment of the present invention, baicalein or a derivative thereof at an appropriate concentration is co-incubated with exosomes, wherein the final concentration of baicalein or a derivative thereof in the incubation system is 5-500 μg / mL, for example, 50-300 μg / mL, calculated as baicalein, and the final concentration of exosomes is 2-3000 μg / mL, for example, 50-200 μg / mL; and the incubation conditions are 0°C-5°C for 1-25 hours, or 20°C-35°C for 1-4 hours.

[0019] In one embodiment of the present invention, an appropriate concentration of baicalein is co-incubated with exosomes; preferably, in the incubation system, the final concentration of baicalein is 5-500 μg / mL (e.g., 6 μg / mL, 7 μg / mL, 8 μg / mL, 9 μg / mL, 10 μg / mL, 20 μg / mL, 30 μg / mL, 40 μg / mL, 45 μg / mL, 50 μg / mL, 55 μg / mL, 60 μg / mL, 70 μg / mL, 80 μg / mL, 90 μg / mL, 100 μg / mL, 110 μg / mL, 120 μg / mL, 130 μg / mL, 140 μg / mL, 150 μg / mL, 160 μg / mL, 170 μg / mL, 180 μg / mL). mL, 190μg / mL, 200μg / mL, 210μg / mL, 220μg / mL, 230μg / mL, 240μg / mL, 250μg / mL, 260μg / mL, 270μg / mL, 280μg / mL, 290μg / mL, 300μg / mL, 310μg / mL, 320μg / m L, 330μg / mL, 340μg / mL, 350μg / mL, 360μg / mL, 370μg / mL, 380μg / mL, 390μg / mL, 400μg / mL, 410μg / mL, 420μg / mL, 430μg / mL, 440μg / mL, 450μg / mL, 460μg / mL, 470 μg / mL, 480 μg / mL, 490 μg / mL, 495 μg / mL, etc.), for example, it can be 50-300 μg / mL,The final concentration of exosomes is 2-3000 μg / mL (e.g., 3 μg / mL, 4 μg / mL, 5 μg / mL, 6 μg / mL, 7 μg / mL, 8 μg / mL, 9 μg / mL, 10 μg / mL, 20 μg / mL, 30 μg / mL, 40 μg / mL, 45 μg / mL, 50 μg / mL, 55 μg / mL, 60 μg / mL, 70 μg / mL, 80 μg / mL, 90 μg / mL, 100 μg / mL). mL, 110μg / mL, 120μg / mL, 130μg / mL, 140μg / mL, 150μg / mL, 160μg / mL, 170μg / mL, 180μg / mL, 190μg / m L, 200μg / mL, 210μg / mL, 220μg / mL, 230μg / mL, 240μg / mL, 250μg / mL, 260μg / mL, 270μg / mL, 280μg / mL, 290μg / mL, 300μg / mL, 350μg / mL, 400μg / mL, 450μg / mL, 500μg / mL, 550μg / mL, 600μg / mL, 650μg / mL, 7 00μg / mL, 750μg / mL, 800μg / mL, 850μg / mL, 900μg / mL, 950μg / mL, 1000μg / mL, 1500μg / mL, 2000μg / mL, 2500μg / mL, 2600μg / mL, 2700μg / mL, 2800μg / mL, 2900μg / mL, 2950μg / mL, 2960μg / mL, 2970μg / mL, 2980μg / mL, 2990μg / mL, 2995μg / mL, etc.), for example, it can be 50-200μg / mL; more preferably, the incubation conditions are 0℃-5℃, 1-25h, or, 20℃-35℃, 1-4h.

[0020] In one embodiment of the present invention, the ultrasonic method comprises: mixing an exosome suspension with a solution of baicalein or a derivative thereof, and ultrasonicating the mixed solution using an ultrasonic instrument to obtain the exosome composition containing baicalein or a derivative thereof.

[0021] In one embodiment of the present invention, the freeze-thaw method comprises: mixing an exosome suspension and a solution of baicalein or a derivative thereof, freezing the mixture in a refrigerator, and then thawing and refreezing the mixture several times to obtain the exosome composition containing baicalein or a derivative thereof.

[0022] A fourth aspect of the present invention provides a pharmaceutical composition comprising the exosome composition described in the first aspect of the present invention, or further comprising a pharmaceutically acceptable excipient. The present invention also provides a pharmaceutical composition for treating rhinosinusitis, comprising the exosome composition described in the first aspect of the present invention, or further comprising a pharmaceutically acceptable excipient.

[0023] In one embodiment of the present invention, only a portion of the baicalein or its derivatives in the pharmaceutical composition is loaded into exosomes. In one embodiment of the present invention, all of the baicalein or its derivatives in the pharmaceutical composition are loaded into exosomes. According to the present invention, the pharmaceutical composition can be in various dosage forms known in the art, such as liquid preparations (including but not limited to suspensions), solid preparations (including but not limited to tablets, powder injections, etc.), and semi-solid preparations (including but not limited to ointments).

[0024] In one embodiment of the present invention, the pharmaceutical composition is a suspension, which, in addition to containing an exosome composition comprising baicalein or a derivative thereof, may further contain a buffer, which may be a buffer commonly used in the art, including but not limited to PBS buffer, DPBS buffer, Tris buffer, Hepes buffer, sodium phosphate, sodium citrate, sodium succinate, histidine (or histidine-HCl), sodium malate, sodium carbonate, etc.

[0025] In one embodiment of the present invention, the pharmaceutical composition is a lyophilized preparation, which, in addition to containing the exosome composition comprising baicalein or its derivatives, may further contain a cryoprotectant. The cryoprotectant may be a cryoprotectant commonly used in the art, including but not limited to mannitol, sucrose, trehalose, lactose, glycerol, dextrose, etc. For lyophilized preparations, water for injection or buffer may be added to reconstitute the suspension before use.

[0026] According to the present invention, in the pharmaceutical composition, the content of exosomes and the content of baicalein or its derivatives (calculated as baicalein) can be independently 0.1-500 μg, for example, 0.2 μg, 0.3 μg, 0.4 μg, 0.5 μg, 0.6 μg, 0.7 μg, 0.8 μg, 0.9 μg, 1 μg, 2 μg, 3 μg, 4 μg, 5 μg, 6 μg, 7 μg, 8 μg, 9 μg, 10 μg, 20 μg, 30 μg, 40 μg, 50 μg, 60 μg, 70 μg, 80 μg, 90 μg, 100 μg, 110 μg, 120 μg, 130 μg, 140 μg, respectively. μg, 150μg, 160μg, 170μg, 180μg, 190μg, 200μg, 210μg, 220μg, 230μg, 240μg, 250μg, 260μg, 270μg, 280μg, 290μg, 300μg, 310μg, 320μg , 330μg, 340μg, 350μg, 360μg, 370μg, 380μg, 390μg, 400μg, 410μg, 420μg, 430μg, 440μg, 450μg, 460μg, 470μg, 480μg, 490μg, 495μg, etc.When the pharmaceutical composition is a suspension, the concentration of the exosomes and the concentration of baicalein or its derivatives calculated as baicalein can be independently 2-500 μg / mL, for example, 2.1 μg / mL, 2.5 μg / mL, 3 μg / mL, 4 μg / mL, 5 μg / mL, 6 μg / mL, 7 μg / mL, 8 μg / mL, 9 μg / mL, 10 μg / mL, 20 μg / mL, 30 μg / mL, 40 μg / mL, 50 μg / mL, 60 μg / mL, 70 μg / mL, 80 μg / mL, 90 μg / mL, 100 μg / mL, 110 μg / mL, 120 μg / mL, 130 μg / mL, 140 μg / mL, 150 μg / mL, 160 μg / mL, 170 μg / mL, 180 μg / mL, 190 μg / mL. g / mL, 200μg / mL, 210μg / mL, 220μg / mL, 230μg / mL, 240μg / mL, 250μg / mL, 260μg / mL, 270 μg / mL, 280μg / mL, 290μg / mL, 300μg / mL, 310μg / mL, 320μg / mL, 330μg / mL, 340μg / mL, 35 0μg / mL, 360μg / mL, 370μg / mL, 380μg / mL, 390μg / mL, 400μg / mL, 410μg / mL, 420μg / mL, 430μg / mL, 440μg / mL, 450μg / mL, 460μg / mL, 470μg / mL, 480μg / mL, 490μg / mL, 495μg / mL, etc. A fifth aspect of the present invention is to provide use of the pharmaceutical composition in the preparation of a medicament for treating rhinosinusitis.

[0027] According to the present invention, the exosome composition comprising baicalein or its derivatives can be administered by various administration methods known in the art, such as oral administration, nasal administration, subcutaneous injection, topical administration, or intravenous administration, etc. In a preferred embodiment, nasal administration is adopted. The dosage of exosomes and the dosage of baicalein or its derivatives calculated as baicalein are independently 0.1-500 μg, for example, 0.2 μg, 0.3 μg, 0.4 μg, 0.5 μg, 0.6 μg, 0.7 μg, 0.8 μg, 0.9 μg, 1 μg, 2 μg, 3 μg, 4 μg, 5 μg, 6 μg, 7 μg, 8 μg, 9 μg, 10 μg, 20 μg, 30 μg, 40 μg, 50 μg, 60 μg, 70 μg, 80 μg, 90 μg, 100 μg, 110 μg, 120 μg, 130 μg, 140 μg, 150 μg, 160 μg, 170 μg, 180 μg, 190 μg, 200 μg, 210 μg, 220 μg, 230 μg, 240 μg, 250 μg, 260 μg, 270 μg, 280 μg, 290 μg, 300 μg, 310 μg, 320 μg, 330 μg, 340 μg, 350 μg, 360 μg, 370 μg, 380 μg, 390 μg, 400 μg, 410 μg, 420 μg, 430 μg, 440 μg, 450 μg, 460 μg, 470 μg, 480 μg, 490 μg, 500 μg, 510 μg, 510 μg, 520 μg, 530 μ 60μg, 170μg, 180μg, 190μg, 200μg, 210μg, 220μg, 230μg, 240μg, 250μg, 260μg, 270μg, 280μg, 290μg, 300μg, 310μg, 320μg, 330μ g, 340μg, 350μg, 360μg, 370μg, 380μg, 390μg, 400μg, 410μg, 420μg, 430μg, 440μg, 450μg, 460μg, 470μg, 480μg, 490μg, 495μg, etc.

[0028] A sixth aspect of the present invention is to provide a method for treating a disease, comprising administering an effective amount of the exosome composition comprising baicalein or a derivative thereof, or the pharmaceutical composition, of the present invention to a patient in need of treatment. According to the present invention, the disease is rhinosinusitis.

[0029] The present invention also provides the use of the exosome composition comprising baicalein or its derivatives or the pharmaceutical composition of the present invention in the treatment of rhinosinusitis.

[0030] The present invention also provides use of the exosome composition comprising baicalein or its derivatives or the pharmaceutical composition of the present invention in the treatment of rhinosinusitis.

[0031] According to the present invention, the baicalein or its derivatives are selected from one or more of the group consisting of baicalein, a pharmaceutically acceptable salt of baicalein, baicalin and a pharmaceutically acceptable salt of baicalin.

[0032] According to the present invention, the pharmaceutically acceptable salts of baicalein and baicalin can be selected from salts of organic acids and salts of inorganic acids, respectively. The inorganic acid can be hydrochloric acid, sulfuric acid, phosphoric acid, diphosphoric acid, hydrobromic acid, or nitric acid. The organic acid can be acetic acid, maleic acid, fumaric acid, tartaric acid, succinic acid, lactic acid, p-toluenesulfonic acid, salicylic acid, or oxalic acid.

[0033] In some embodiments of the present invention, the rhinosinusitis is chronic rhinosinusitis, preferably eosinophilic chronic sinusitis, with or without nasal polyps.

[0034] According to the present invention, the treatment includes reducing or alleviating or inhibiting inflammation of the mucous membrane of the nasal cavity and / or paranasal sinuses, repairing the mucous membrane of the nasal cavity and / or paranasal sinuses, and inhibiting the thickness of the mucous membrane of the nasal cavity and / or paranasal sinuses.

[0035] According to the present invention, the exosomes may be exosomes from human or mammalian cells.

[0036] The mammals include but are not limited to ungulates such as cattle, sheep, horses, and pigs, rodents such as rabbits, rats, mice, and guinea pigs, primates such as monkeys and gorillas, cats, dogs, etc. In one embodiment of the present invention, the exosomes are exosomes of human cells.

[0037] The cells may be normal somatic cells and / or stem cells, including but not limited to vascular endothelial cells, dendritic cells, T cells (e.g., Foxp3 + Treg cells), neurons, oligodendrocytes, astrocytes, microglia, embryonic stem cells, adult stem cells (e.g., hematopoietic stem cells, neural stem cells, mesenchymal stem cells derived from bone marrow, umbilical cord, fat, umbilical cord blood, amniotic membrane, (placental) chorionic villi, dental pulp, thymus, synovium, etc.), etc.

[0038] Induced pluripotent stem cells (iPSCs) technology can also be used to reprogram terminally differentiated somatic cells into pluripotent stem cells by introducing specific genes (such as Oct4, Sox2, Klf4, c-Myc, etc.). iPSCs can then be used to obtain exosomes derived from iPSCs. In this invention, iPSCs exosomes are abbreviated as "iPS-EVs." iPSCs technology can be used to prepare proprietary induced pluripotent stem cells from a patient's own somatic cells. Exosomes are then obtained from the patient's proprietary induced pluripotent stem cells and used on the patient's own body, significantly reducing the possibility of immune rejection.

[0039] In some embodiments of the present invention, the exosomes are exosomes derived from pluripotent stem cells. In some embodiments of the present invention, the exosomes are exosomes derived from induced pluripotent stem cells, exosomes derived from mesenchymal stem cells, or exosomes derived from embryonic pluripotent stem cells. In some embodiments of the present invention, the exosomes are human pluripotent stem cell exosomes, for example, exosomes derived from human induced pluripotent stem cells, exosomes derived from human mesenchymal stem cells, or exosomes derived from human embryonic pluripotent stem cells.

[0040] Exosomes can be isolated and prepared from the cell culture medium by culturing the corresponding cells in vitro through autocrine secretion. Each cell type can be cultured using methods and culture media known in the art. For example, the culture medium for human skin fibroblasts can be DMEM + 15% FBS + NEAA, the culture medium for umbilical cord mesenchymal stem cells can be DMEM + 10% FBS, and the culture medium for neuronal cells can be B27 + Neurobasal + GlutaMAX.

[0041] To increase the production of exosomes in cell culture, the content of chemicals in the culture medium can be adjusted according to the characteristics of the cultured cells. For example, the chemicals essential for cell growth can be retained in the culture medium, while chemicals not essential for cell growth or their content can be removed or reduced, thereby placing the cells in a "starvation" state to increase the production of exosomes in the culture. For example, to culture stem cells to obtain stem cell exosomes, L-ascorbic acid or its salts, selenium or its salts, insulin, etc. can be added to the stem cell basal culture medium, such as DMEM, F12, or DMEM / F12, to induce the stem cells to secrete large amounts of exosomes. For example, the exosome secretion inducer described in Chinese patent application CN112920991A is incorporated into this application in its entirety.

[0042] In the present invention, various exosome isolation methods in the art can be used to isolate and obtain exosomes from cell culture fluid.

[0043] Differential ultracentrifugation is the most commonly used method for isolating and concentrating exosomes. Differential ultracentrifugation, which uses different centrifugal forces to precipitate different impurities in the sample, allows for the extraction of purer exosomes. This method can be combined with ultrafiltration using a 0.22μm or 0.45μm pore size membrane to improve product purity and reduce exosome aggregation.

[0044] Exosomes are a subpopulation of extracellular vesicles with a diameter ranging from 30 to 150 nm. Exosomes can also be separated based on particle size. Methods include ultrafiltration, size exclusion chromatography, hydrostatic filtration dialysis, and asymmetric field flow fractionation.

[0045] Commercial exosome extraction kits such as Exo-spin can also be used TM 、ExoQuick TM 、Invitrogen TM and other products to isolate exosomes.

[0046] The extracted exosomes can be suspended in a buffer, such as phosphate buffer, and stored frozen at -80°C to 4°C. They can also be stored in a lyophilized form by adding a cryoprotectant, such as trehalose.

[0047] Definition of terms:

[0048] “And / or” will be taken as a specific disclosure of each of the two specified features or components with or without the other. Thus, the term “and / or” used in phrases such as “A and / or B” is intended to include “A and B,” “A or B,” “A” (alone), and “B” (alone). Likewise, the term “and / or” used in phrases such as “A, B, and / or C” is intended to cover each of the following: A, B, and C; A, B, or C; A or C; A or B; B or C; A and C; A and B; B and C; A (alone); B (alone); and C (alone).

[0049] "Comprising" and "including" have the same meaning and are intended to be open ended and permit but not require the inclusion of additional elements or steps. When the terms "comprising" or "including" are used herein, the terms "consisting of" and / or "consisting essentially of" are also included and disclosed.

[0050] "Pharmaceutically acceptable excipients" refers to any ingredient other than the exosomes described herein that is substantially non-toxic and non-inflammatory in a patient, including, but not limited to, any and all solvents, dispersion media or other liquid carriers, dispersion or suspension aids, diluents, isotonicity agents, preservatives, colorants, sweeteners or flavorings, stabilizers, antioxidants, antimicrobial or antifungal agents, osmolarity adjusters, pH adjusters, buffers, chelating agents, cryoprotectants, and / or fillers, as appropriate for the particular dosage form desired. Various excipients for formulating pharmaceutical compositions and techniques for preparing compositions are known in the art. Exemplary antimicrobial or antifungal agents include, but are not limited to, benzalkonium chloride, benzethonium chloride, methylparaben, ethylparaben, propylparaben, butylparaben, benzoic acid, hydroxybenzoic acid, potassium or sodium benzoate, potassium or sodium sorbate, sodium propionate, sorbic acid, and the like, and combinations thereof. Exemplary preservatives include, but are not limited to, vitamin A, vitamin C, vitamin E, β-carotene, citric acid, ascorbic acid, and the like, and combinations thereof. Exemplary buffers for controlling pH may include, but are not limited to, sodium phosphate, sodium citrate, sodium succinate, histidine (or histidine-HCl), sodium malate, sodium carbonate, and the like, and / or combinations thereof. Exemplary cryoprotectants include, but are not limited to, mannitol, sucrose, trehalose, lactose, glycerol, dextrose, and the like, and combinations thereof. BRIEF DESCRIPTION OF THE DRAWINGS

[0051] Figure 1: Standard curve of baicalein concentration and absorbance.

[0052] Figure 2: H&E stained sections of mouse sinus tissue.

[0053] Figure 3: Statistical graph of the thickness of the olfactory epithelium and nasal mucosa in the mouse sinus tissue.

[0054] Figure 4: Fluorescence immunohistochemistry of ciliated cells in mouse sinus tissue.

[0055] Figure 5: Fluorescence immunohistochemistry of basal cells in mouse sinus tissue. DETAILED DESCRIPTION

[0056] The present invention is further described below with reference to the following examples. It should be noted that the examples are not intended to limit the scope of protection of the present invention, and those skilled in the art will understand that any improvements and variations based on the present invention are within the scope of protection of the present invention.

[0057] The conventional reagents used in the following examples are all commercially available. The biological experiments performed are conventional biological experiments in the art and can be performed according to the corresponding experimental manuals or kit instructions.

[0058] Example 1 Preparation of exosomes from human induced pluripotent stem cells

[0059] (1) Cell culture: iPSC cells (product of Guodian (Beijing) Pharmaceutical Technology Co., Ltd.) were expanded and cultured (using the Essential 8™ Medium kit (purchased from Thermo Fisher, product number: A1517001) for routine expansion culture) until the logarithmic growth phase and in good condition. According to the technical manual of the Versene Solution kit (purchased from Thermo Fisher, product number: 15040066), they were digested into small cell clusters (3-10 cells), and then plated on a 10 cm cell culture dish coated with vitronectin (Recombinant Human Vitronectin purchased from Peprotech, product number 140-09). After adhesion, the coverage rate reached 60% to 70%. Cultured overnight at 37°C in an atmosphere of 95% air and 5% CO2. When the iPSCs were in good condition, the medium was replaced with GDEV medium (product of Guodian (Beijing) Pharmaceutical Technology Co., Ltd.). After 24 hours, the medium was replaced and the cell culture supernatant was collected. The previous step was repeated. A batch of cells can be collected 3 times.

[0060] (2) Isolation of exosomes: The cell culture supernatant obtained from the induced culture of cells in step (1) was centrifuged at 3000 g for 15 min at room temperature to remove dead cells and cell debris. The supernatant was collected and filtered through a 0.22 μm filter; the filtrate was transferred to a Centricon Plus-70 (100 kDa) ultrafiltration tube and concentrated, and centrifuged at 3000 g for 30 min at room temperature. The concentrate was diluted with DPBS (Dulbecco's phosphate buffered saline) at a ratio of approximately 1:100 and then concentrated again using the same device to obtain an exosome solution.

[0061] Example 2 Preparation of exosomes from human umbilical cord mesenchymal stem cells

[0062] (1) Sample processing: 10 cm of fresh umbilical cord tissue was collected and washed with normal saline containing penicillin-streptomycin solution (double antibody) (100×) (purchased from Gibco) and cut into 1 mm pieces with ophthalmic scissors. 2 The small pieces were transferred to cell culture flasks, and DMEM / F12 culture medium containing 10% serum replacement and penicillin-streptomycin (both purchased from Gibco) was added, and the cells were cultured in a 5% CO2, 37°C incubator.

[0063] (2) Cell culture: During the primary culture in step (1), the medium was changed for the first time after 1 week, and then every 3-4 days. When the cells reached 80%-90% confluency, they were digested with trypsin / EDTA solution (purchased from Gibco) and transferred to new culture dishes for culture.

[0064] (3) Isolation of exosomes: When the cell concentration grows to 80-90%, human mesenchymal stem cell serum-free culture medium (MSC XF Medium, purchased from Biological Industries), and continued culturing. After 24 h, the medium was replaced and the cell culture supernatant was collected. The previous step was repeated, and one batch of cells could be collected three times. The cell culture supernatant collected three times was centrifuged for 20 min (3000 g / min) to remove cells and cell debris, and then centrifuged at 3000 g / min for 30 min at room temperature using Centricon Plus-70 (100 kDa) ultrafiltration tubes (purchased from Sigma) to obtain exosomes.

[0065] Example 3 Preparation of exosome composition containing baicalein

[0066] 1.1 Weigh 5 mg of baicalein powder and add 1.15 mL of DMSO to obtain a 16 mM DMSO stock solution. Aliquot into 10 tubes and store in a -80°C refrigerator protected from light.

[0067] 1.2 Take out the sample with a density of 10 from the -80℃ refrigerator 11 / mL (density is 10 11 Thaw 100 μL of iPSC exosomes (exosome concentration of approximately 1 mg / mL) at room temperature, add 290 μL of PBS solution, and then add 10 μL of the 16 mM baicalein DMSO solution from step 1.1. At this point, the concentration of baicalein in the system is 0.4 mM (approximately 108 μg / mL), and the system contains 2.5% DMSO. Vortex to mix, and place in the dark at room temperature for 2 h, or in a refrigerator at 4°C overnight.

[0068] 1.3 Transfer the solution to a 100K ultrafiltration tube, set the centrifugal force to 14,000 × g, and centrifuge for 5 minutes.

[0069] 1.4 Add 400 μL of PBS solution containing 2.5% DMSO to the ultrafiltration tube and centrifuge at 14,000 × g for 5 minutes. Repeat this washing step four times and collect the final wash solution.

[0070] 1.5 Add 400 μL of PBS solution without DMSO, set the centrifugal force to 14,000 × g, and centrifuge for 5 minutes.

[0071] 1.6 Carefully aspirate the drug-loaded exosomes from the ultrafiltration tube using a 20 μL pipette and transfer them to a 0.2 mL centrifuge tube. Store in a -80°C freezer.

[0072] Example 4 Detection of the drug loading amount of baicalin in the exosome composition containing baicalin

[0073] 1.1. Preparation of baicalein standard solution

[0074] 1) Take 10 μL of a 16 mM baicalein DMSO solution and add 10 μL of DMSO to obtain an 8 mM baicalein DMSO solution.

[0075] 2) Take 10 μL of the above 8 mM baicalein solution and add it to 390 μL of PBS solution to obtain a standard solution with a concentration of 200 μM.

[0076] 3) Take 200 μL of the above 200 μM standard solution and add 200 μL of PBS solution containing 2.5% DMSO to obtain a standard solution with a concentration of 100 μM.

[0077] 4) According to the above dilution method, dilute in sequence to obtain standard solutions with concentrations of 50 μM, 25 μM, 10 μM, 5 μM, 1 μM, and 0 μM.

[0078] 1.2 UV-visible absorbance spectrophotometry using an enzyme-labeled instrument

[0079] 1) Prepare standard curve solution

[0080] Take 4.5 μL of each standard solution prepared in 1.1 and place it in a 0.2 mL centrifuge tube. 11 3 μL of iPSC exosomes at a concentration of 1 μg / mL. The gradient concentration of baicalein in the system was 120 μM, 60 μM, 30 μM, 15 μM, 6 μM, 3 μM, 0.6 μM, and 0 μM (blank control). All solutions contained 1.5% DMSO.

[0081] 2) Dilute the sample solution

[0082] 3 μL of the exosome composition containing baicalein prepared in Example 3 was taken as a sample, and 4.5 μL of a PBS solution containing 2.5% DMSO was added. The DMSO content at this point was 1.5%.

[0083] 3) Add 6 μL of each of the standard curve solution and diluted sample solution (three replicate wells) to a microplate. Measure the OD value in an M5 microplate reader at a wavelength of 338 nm. The absorbance values ​​for the three replicate wells of the sample solution are shown in Table 1.

[0084] 4) In Origin8.0 software, the concentration of the standard curve solution was used as the horizontal axis and the OD value after deducting the blank absorption was used as the vertical axis to perform linear fitting and obtain the linear equation and R 2 The values ​​are shown in Figure 1.

[0085] 5) Calculate the concentration of baicalein in the diluted sample according to the standard curve equation. Multiply this concentration by 2.5 to obtain the loading concentration of baicalein in the exosome composition containing baicalein prepared in Example 3.

[0086] Table 1 Absorbance values ​​of 3 parallel wells of diluted sample solution

[0087] Substituting the absorbance values ​​of the three replicate wells of the diluted sample solution into the standard curve (y = 0.0016x), the concentrations of baicalein in the diluted sample were found to be 20 μM, 19.13 μM, and 19.81 μM, with an average concentration of 19.65 μM. Therefore, the loading concentration of baicalein in the exosome composition containing baicalein prepared in Example 3 was 19.65 μM × 2.5 = 49.12 μM.

[0088] The drug loading rate of baicalein % = 49.12 μM / 400 μM×100% = 12.28%.

[0089] The applicant used the above method to change the incubation conditions, different final concentrations of exosomes in the incubation system, and different baicalein concentrations, and measured the loading concentration of baicalein in the exosome composition containing baicalein. The results are as follows:

[0090] (1) PBS system, containing 2.5×10 8 exosomes / mL, 200 μM baicalein

[0091] (2) PBS system, containing 3.0×10 11 exosomes / mL, 1mM baicalein

[0092] Example 5: Treatment of Eosinophilic Sinusitis with an Exosome Composition Containing Baicalein

[0093] Eosinophilic sinusitis modeling protocol in mice: Weigh an appropriate amount of papain and dissolve it in PBS to a final concentration of 1 mg / ml. Instill 5 μL of papain solution into the nostrils of each mouse, alternating morning and evening, for modeling, followed by 20 μL per mouse per day.

[0094] Grouping scheme: Mice were induced with an eosinophilic rhinitis model using papain intranasally (days 0-2 and 7-10), and then divided into a PBS group, a PC (positive control) group, an EXO-1 (exosome) group, a baicalein (hereinafter abbreviated as BC) group, and a BC+EXO-1 (exosome composition containing baicalein) group and administered the drugs respectively.

[0095] Other normal mice were induced with physiological saline (days 0-2 and 7-10) as a sham normal control group.

[0096] From day 7 to day 13 after modeling, the sham group, PBS group, PC group, EXO-1 group, BC group, and BC+EXO-1 group were intervened by intranasal drops of PBS, PBS, Nasonex, iPSC exosomes, baicalein, and an exosome composition containing baicalein in the morning and evening, respectively. On day 14, the sinus tissues of the mice were collected, embedded in paraffin, and stained with hematoxylin and eosin to observe the nasal mucosa.

[0097] Group setting and drug preparation instructions

[0098] 1) Sham group: normal control group, model was established using normal saline and PBS was administered;

[0099] 2) PBS group: placebo group, using papain-induced rhinitis model, PBS administration;

[0100] 3) PC group: positive control group, using papain-induced rhinitis model, administered with Nasonex (Nasonex: mometasone furoate nasal spray), 10 μL / time, once in the morning and evening;

[0101] 4) EXO-1 group: Exosome treatment group, using papain-induced rhinitis model, iPSC exosomes were administered at a final concentration of 100 μg / ml, 10 μL / time (exosome content 1 μg), once in the morning and evening;

[0102] 5) BC group: Baicalein treatment group, using papain-induced rhinitis model, baicalein at a concentration of 100 μg / mL (in 2% DMSO as solvent), 1 μg / time (about 10 μL / time) intranasal drops, once in the morning and evening;

[0103] 6) BC+EXO-1 Group: Treatment with an exosome composition containing baicalein. Using a papain-induced rhinitis model, baicalein at a final concentration of 100 μg / mL (in 2% DMSO) was co-incubated with 100 μg / mL iPSC exosomes (20 h at 4°C or 3 h at room temperature). The mixture was then administered intranasally at a concentration of 10 μL (1 μg exosomes and 1 μg baicalein) once in the morning and evening.

[0104] Experimental procedures

[0105] 1) Thirty male C57BL / 6J mice weighing 18-22 g were acclimated for one week and then numbered according to the table below and randomly divided into six groups of five mice each.

[0106] 2) Grouping was performed according to the table below. On days 0-2 and 7-10, groups 2-6 (PBS group and multiple treatment groups) received alternating intranasal papain drops (20 μL / mouse). Group 1 (sham group) received intranasal drops of an equal volume of normal saline.

[0107] 3) On days 7-13, groups 3-6 received alternating intranasal drops of PC, EXO-1, BC, and BC + EXO-1, respectively, while groups 1 (sham group) and 2 (PBS group) received intranasal drops of an equal volume of PBS solution;

[0108] 4) On day 14, the sinus tissues of the mice were collected, fixed, and embedded in paraffin. Subsequently, HE staining was performed to observe the morphology. The thickness of the nasal mucosa was calculated at three locations on each section, and the average of the three locations on each section was used for statistical analysis.

[0109] Animal experiment groups:

[0110] Experimental results

[0111] Epithelial cell damage was assessed using H&E-stained sections. As shown in Figure 2, the nasal mucosa in the sham group showed intact olfactory epithelial mucosa and intact cilia. In the PBS group, the nasal mucosa was severely damaged, with atrophy of the olfactory epithelium, loose and damaged cells, pyknotic and darkly stained nuclei in some cells, vacuolar changes, and cilia loss. In the PC group, the nasal mucosa showed atrophy of the olfactory epithelium, loose and damaged cells, pyknotic and darkly stained nuclei in some cells, and vacuolar changes, but some cilia collapsed. In the BC group, the nasal mucosa showed intact olfactory epithelial mucosa, but cilia loss. In the EXO-1 group, the nasal mucosa showed partial cilia loss. In the BC+EXO-1 group, the nasal mucosa showed intact olfactory epithelial mucosa and intact cilia. H&E-stained sections show that the nasal mucosa in the BC+EXO-1 group was intact and had no cilia loss, demonstrating reduced nasal mucosal damage compared to the PBS group, the PC group, and the groups receiving BC or EXO-1 alone.

[0112] Nasal mucosal thickening was assessed using H&E-stained sections. Nasal mucosal thickening typically indicates chronic inflammation in the nasal cavity, resulting from recurrent rhinitis or long-term chronic local irritation. As shown in Figure 3, in the saline-treated (Sham) group, the nasal mucosal thickness was approximately 30 μm. In the PBS group, due to edema caused by the continuous stimulation of inflammation, the nasal mucosa thickened to approximately 40 μm. Neither the Nasonex (PC) group nor the baicalein-containing (BC) group showed significant inhibition of nasal mucosal thickening compared to the PBS group. The overall average thickness of the nasal mucosa in the exosome EXO-1 group decreased compared to the PBS group, but the difference was not significant. The baicalein-containing exosome combination BC+EXO-1 group had the greatest therapeutic effect. After treatment, the nasal mucosal thickness of mice in the BC+EXO-1 group reached the same level as the Sham group, indicating that exosomes containing baicalein can effectively inhibit the inflammatory response of the nasal mucosa and have a good therapeutic effect on chronic sinusitis.

[0113] Example 6: Treatment of Eosinophilic Sinusitis with an Exosome Composition Containing Baicalin

[0114] A mouse model of eosinophilic sinusitis was constructed and the experiment was conducted according to the same experimental method as in Example 5, except that baicalin was used instead of baicalein, and the dosage concentration of the baicalin treatment group was adjusted to 200 μg / mL of baicalin (121 μg / mL as baicalein). When preparing the encapsulated exosomes, the final concentration of baicalin in the incubation system was 200 μg / mL (121 μg / mL as baicalein).

[0115] After obtaining mouse sinus tissue, paraffin embedding, H&E staining for histological observation, immunofluorescence staining (using β-tubulin and CK-5, respectively), and quantitative analysis of the integrity of ciliated cells and the proliferation of basal cells in sinus tissue were performed. The immunofluorescence intensity of ciliated cells per unit length was calculated at 8 locations on each section to assess the integrity / loss of ciliated cells. The immunofluorescence intensity of ciliated cells per unit length at 8 locations on each section was averaged for data statistics. The immunofluorescence intensity of basal cells per unit length was calculated at 6 locations on each section to assess basal cell proliferation. The immunofluorescence intensity of basal cells per unit length at 6 locations on each section was averaged for data statistics.

[0116] The experimental results for ciliated cells are shown in Figure 4. Figure 4 (a) shows the immunofluorescence intensity of ciliated cells per unit length in each section, which was used to assess the integrity / loss of ciliated cells at eight selected locations. Figure 4 (b) shows the results of immunofluorescence staining of sections. It can be seen that the fluorescence expression of ciliated cells in the PC group (Nasonex treatment group) and the NS group (normal saline group) was relatively weak, while the fluorescence expression of ciliated cells in the BC+EXO group (exosomes containing baicalin) was stronger than that in the EXO group (exosomes alone) and the BC group (baicalin alone).

[0117] Figure 4(c) shows the quantitative statistical results of the immunofluorescence intensity of ciliated cells per unit length. Stronger immunofluorescence intensity indicates more intact ciliated cells and less ciliated cell shedding. Conversely, weaker immunofluorescence intensity indicates greater damage to ciliated cells and more ciliated cell shedding. Figure 4(c) shows that the immunofluorescence intensity of ciliated cells per unit length in the PC group was the weakest. The immunofluorescence intensity of ciliated cells per unit length in the EXO and BC groups was stronger than that in the NS group, but there was no significant difference compared with the NS group. The immunofluorescence intensity of ciliated cells per unit length in the EXO+BC group was the strongest and significantly different from the NS group. This suggests that treatment with exosomes encapsulating baicalin can significantly reduce ciliated cell shedding and maintain ciliated cell integrity, with a better effect than that of nasonex or exosomes or baicalin alone.

[0118] The results for basal cells are shown in Figure 5. Figure 5(a) shows the immunofluorescence intensity of basal cells per unit length in each section, which was used to assess basal cell proliferation at six selected locations. Figure 5(b) shows the immunofluorescence staining results for the sections. As can be seen, the fluorescence expression of basal cells in the NS group was relatively strong, indicating a stronger inflammatory response.

[0119] Figure 5(c) shows the quantitative statistical results of the immunofluorescence intensity of basal cells per unit length. Stronger immunofluorescence intensity indicates more basal cell proliferation and more severe inflammation. Conversely, weaker immunofluorescence intensity indicates less basal cell proliferation and milder inflammation. As shown in Figure 5(c), the immunofluorescence intensity of basal cells per unit length in the PC group is similar to that in the Sham group. The immunofluorescence intensity of basal cells per unit length in the EXO and BC groups is slightly higher than that in the Sham group and lower than that in the NS group, but there is no significant difference. The immunofluorescence intensity of basal cells per unit length in the BC+EXO group is weaker than that in the EXO, BC, and NS groups, and is significantly different from that in the NS group. This indicates that treatment with exosomes encapsulating baicalin can significantly reduce the inflammatory response and basal cell proliferation, which is better than using exosomes or baicalin alone, and also better than using nasonex.

[0120] The above describes the embodiments of the present invention. However, the present invention is not limited to the above embodiments. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principles of the present invention shall be included in the scope of protection of the present invention.

Claims

1. An exosome composition comprising baicalein or a derivative thereof, characterized in that: A composition comprising baicalein or a derivative thereof and exosomes as active ingredients; only a portion of the baicalein or a derivative thereof in the exosome composition is loaded in the exosomes, or all of the baicalein or a derivative thereof in the exosome composition is loaded in the exosomes; the loading concentration of the baicalein or a derivative thereof is 2-500 μg / mL in terms of baicalein; Calculated as baicalein, in the exosome composition, the amount of baicalein or its derivatives loaded in the exosomes accounts for 5%-100% of the total amount of baicalein or its derivatives in the exosome composition; The baicalein or its derivative is selected from one or more of the group consisting of baicalein, a pharmaceutically acceptable salt of baicalein, baicalin and a pharmaceutically acceptable salt of baicalin.

2. The exosome composition according to claim 1, characterized in that The exosome composition is obtained by mixing the exosomes with the baicalein or a derivative thereof and incubating the mixture.

3. The exosome composition according to claim 2, characterized in that The incubation comprises: co-incubating an appropriate concentration of baicalein or its derivatives with exosomes; in the incubation system, the final concentration of baicalein or its derivatives is 5-500 μg / mL in terms of baicalein, and the final concentration of exosomes is 2-3000 μg / mL; the incubation conditions are 0°C-5°C, 1-25h, or 20°C-35°C, 1-4h.

4. Use of the exosome composition comprising baicalein or a derivative thereof according to any one of claims 1 to 3 in the preparation of a medicament for treating rhinosinusitis.

5. A pharmaceutical composition, characterized in that A composition comprising the exosomes according to any one of claims 1 to 3.

6. The pharmaceutical composition according to claim 5, characterized in that The pharmaceutical composition is a suspension, which contains a buffer in addition to an exosome composition containing baicalein or a derivative thereof, and the concentration of the exosomes and the concentration of baicalein or a derivative thereof calculated as baicalein are independently 2-500 μg / mL, respectively; or, the pharmaceutical composition is a lyophilized preparation, which contains a cryoprotectant in addition to an exosome composition containing baicalein or a derivative thereof, and the content of the exosomes and the content of baicalein or a derivative thereof calculated as baicalein are independently 0.1-500 μg, respectively.

7. The exosome composition according to any one of claims 1 to 3, the use according to claim 4, or the pharmaceutical composition according to any one of claims 5 to 6, characterized in that: The pharmaceutically acceptable salt of baicalein and the pharmaceutically acceptable salt of baicalin are respectively selected from salts of organic acids and salts of inorganic acids; Preferably, the inorganic acid is hydrochloric acid, sulfuric acid, phosphoric acid, diphosphoric acid, hydrobromic acid or nitric acid; Preferably, the organic acid is acetic acid, maleic acid, fumaric acid, tartaric acid, succinic acid, lactic acid, p-toluenesulfonic acid, salicylic acid or oxalic acid.

8. The exosome composition according to any one of claims 1 to 3, the use according to claim 4, or the pharmaceutical composition according to any one of claims 5 to 6, characterized in that: The exosomes are exosomes from human or mammalian cells; Preferably, the exosomes are exosomes of human cells; preferably, the cells are normal somatic cells or stem cells; Preferably, the exosomes are exosomes derived from pluripotent stem cells; preferably, the pluripotent stem cell exosomes are exosomes derived from induced pluripotent stem cells, exosomes derived from mesenchymal stem cells, or exosomes derived from embryonic pluripotent stem cells; preferably, the pluripotent stem cell exosomes are human pluripotent stem cell exosomes, for example, exosomes derived from human induced pluripotent stem cells, exosomes derived from human mesenchymal stem cells, or exosomes derived from human embryonic pluripotent stem cells.

9. A method for treating rhinosinusitis, characterized in that: Administering an effective amount of the exosome composition according to any one of claims 1 to 3 or the pharmaceutical composition according to any one of claims 5 to 6 to a patient in need of treatment.

10. Use of the exosome composition according to any one of claims 1 to 3 or the pharmaceutical composition according to any one of claims 5 to 6 in the treatment of rhinosinusitis.

11. The use according to claim 4, the method according to claim 9, or the application according to claim 10, characterized in that: The rhinosinusitis is chronic rhinosinusitis; Preferably, the rhinosinusitis is eosinophilic chronic sinusitis; Preferably, the eosinophilic chronic sinusitis is with or without nasal polyps; and / or, Preferably, the treatment is to reduce or alleviate or inhibit inflammation of the mucosa of the nasal cavity and / or paranasal sinuses, repair the mucosa of the nasal cavity and / or paranasal sinuses, and / or inhibit the thickness of the mucosa of the nasal cavity and / or paranasal sinuses.

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