Method for preparing a pharmaceutical mixture, the obtained pharmaceutical mixture and its medical use

Abstract

The present invention belongs to the field of medical technology, and specifically relates to a method for preparing a pharmaceutical mixture, comprising the following steps: (1) providing a soluble salt solution containing zinc ions and optionally calcium ions; (2) providing an alkaline solution containing phosphate ions, bisphosphonate ions and hydroxide ions; (3) mixing the soluble salt solution with the alkaline solution to obtain a mixture. The present invention also relates to the prepared pharmaceutical mixture and its medical use. The pharmaceutical mixture of the present invention can prevent or treat respiratory tract-transmitted viral infections, or prevent or treat diseases or symptoms caused by respiratory tract-transmitted viral infections.

Description

Technical Field

[0001] The invention belongs to the field of medical technology, and specifically relates to a method for preparing a medicinal mixture, and also relates to the prepared medicinal mixture and its medical use. Background Art

[0002] Respiratory viruses include influenza virus, respiratory syncytial virus (RSV), and new coronavirus. Influenza virus is one of the main pathogens that infect humans and animals. It is mainly divided into four types: A, B, C, and D. Human influenza is mainly caused by influenza A virus and influenza B virus. Human influenza caused by influenza virus can be divided into seasonal influenza, pandemic influenza, and animal-borne influenza. Seasonal influenza includes types A, B, and C, which are prevalent every year, especially in winter. Seasonal influenza can cause mild or severe illness, and even cause death in high-risk groups such as people with low immunity, pregnant women, infants and the elderly. Pandemic influenza refers to a new type of influenza that has never been prevalent in the human population before. It is easy to cause a global pandemic. For example, the Spanish pandemic influenza from 1918 to 1919 caused about 20 million to 50 million deaths worldwide. Animal-borne influenza includes avian influenza and swine influenza. Infected people are infected through direct contact with sick animals. Animal-borne influenza has a weak ability to spread among people. Respiratory syncytial virus is an RNA virus that can cause infection in newborns and infants under 6 months old, with an incubation period of about one week. Infants and young children infected with RSV have more severe symptoms, generally rhinitis, pharyngitis, high fever and laryngitis, and further develop bronchiolitis and pneumonia.

[0003] In terms of disease prevention, vaccines are the most effective means for humans to fight against influenza viruses today. Traditional influenza vaccines are trivalent inactivated vaccines. Before each flu season, the WHO predicts in advance the H3N2 and H1N1 subtype A influenza virus strains and one sublineage B influenza virus strain that will be prevalent in the northern and southern hemispheres during that flu season. The three virus strains are combined as the trivalent influenza vaccine for that flu season. However, due to the mutability of influenza viruses and the co-circulation of two sublineages of influenza B viruses, the vaccine strains predicted by the WHO often do not conform to the actual epidemic situation, which greatly reduces the effective protection rate of the vaccine and increases the difficulty of prevention work.

[0004] At present, RSV vaccine development also faces huge challenges. There is no vaccine to prevent its infection in the world. Some patients hospitalized with severe infection can only receive supportive treatment and wait for their own immune system to eliminate the virus. For example, in 1960, the wild-type virus "Bernett strain" isolated from the sputum of volunteers was cultured and grown on grassland monkey kidney cells, inactivated with formalin, and then precipitated with aluminum adjuvant to obtain formalin-inactivated vaccine (FI-RSV); but in clinical trials, vaccination with FI-RSV not only failed to resist RSV infection, but also compared with the placebo group, the infection degree of some subjects in the vaccination group was aggravated; the vaccine evaluation study on the youngest age group (2-7 months old infants) showed that after the subjects were infected with RSV again, 16 of the 20 subjects in the vaccination group (80%) needed hospitalization, while only 1 of the 21 subjects in the control group (5%) needed hospitalization, and even two infants in the vaccination group died due to severe RSV infection. This result has a negative impact on the subsequent development of RSV vaccines.

[0005] Therefore, it is of great significance to develop new drugs that have better preventive effects on respiratory viruses. Summary of the invention

[0006] The present invention provides a method for preparing a medicinal mixture, wherein the prepared medicinal mixture can prevent or treat respiratory tract-transmitted viral infection, or prevent or treat diseases or symptoms caused by respiratory tract-transmitted viral infection. Based on this, the present invention also provides the prepared medicinal mixture and its medical use.

[0007] The first aspect of the present invention relates to a method for preparing a pharmaceutical mixture, comprising the following steps:

[0008] (1) providing a soluble salt solution comprising zinc ions and optionally calcium ions;

[0009] (2) Provide phosphate ions (PO4 3- ), an alkaline solution of bisphosphonate ions and hydroxide ions;

[0010] (3) Mixing the soluble salt solution with the alkaline solution to obtain a mixture.

[0011] In some embodiments of the first aspect of the present invention, in step (3), the alkaline solution is added to the soluble salt solution for mixing.

[0012] In some embodiments of the first aspect of the present invention, in step (3), the alkaline solution is dropped into the soluble salt solution for mixing.

[0013] In some embodiments of the first aspect of the present invention, the method further comprises step (4): sterilizing the mixture to obtain a sterilized product.

[0014] In some embodiments of the first aspect of the present invention, the sterilization treatment is high-pressure sterilization treatment.

[0015] In some embodiments of the present invention, high-pressure sterilization refers to placing the object to be treated in a closed high-pressure sterilization environment, and using high-pressure saturated steam (usually 103.4 kPa steam pressure) to denature the proteins and nucleic acids in the microorganisms, thereby killing the microorganisms.

[0016] In some embodiments of the first aspect of the present invention, the temperature of the sterilization treatment is 100°C to 140°C, for example, 110°C, 115°C, 120°C, 125°C, 130°C.

[0017] In some embodiments of the first aspect of the present invention, the sterilization treatment time is 0.5 to 3 hours, for example, 1 hour, 1.5 hours, 2 hours, or 2.5 hours.

[0018] In some embodiments of the first aspect of the present invention, the method further comprises step (5): adjusting the pH value of the mixture or the sterilized product to 6-7, for example, 6.5-7.

[0019] In some embodiments of the first aspect of the present invention, the pH value is adjusted using an acid solution or a hydroxide (eg, sodium hydroxide) solution.

[0020] In some embodiments of the first aspect of the present invention, the sterilized material after adjusting the pH value is diluted (with water).

[0021] In some embodiments of the first aspect of the present invention, the method has one or more of the following features (A) to (F):

[0022] (A) In step (1), the molar concentration of zinc ions in the soluble salt solution is 20 to 83 mM, for example, 30 mM, 35 mM, 40 mM, 42 mM, 45 mM, 50 mM, 55 mM, 60 mM, 65 mM, 70 mM, 80 mM;

[0023] (B) In step (2), the molar concentration of phosphate ions in the alkaline solution is 20 to 65 mM, for example, 30 mM, 35 mM, 40 mM, 45 mM, 50 mM, 55 mM, 60 mM;

[0024] (C) In step (2), the molar ratio of phosphate ion, diphosphonate ion and hydroxide ion in the alkaline solution is (4-7):1:(3-11), preferably (5-6):1:(4.5-9.3), more preferably (5-6):1:(5-9), for example 5.3:1:5.13, 5.3:1:8.46, 5.3:1:7.69;

[0025] (D) In ​​step (3), the volume ratio of the soluble salt solution to the alkaline solution is 1:(0.5-2), for example, 1:0.8, 1:1, 1:1.5;

[0026] (E) a bisphosphonate ion selected from the group consisting of a zoledronic acid ion and a risedronate ion;

[0027] (F) in step (1), the soluble salt solution contains zinc ions and calcium ions, or the soluble salt solution contains zinc ions but no calcium ions;

[0028] Preferably, the molar ratio of zinc ions to calcium ions in the soluble salt solution is 1:(0.4-3), for example, 1:0.7, 1:1, 1:1.5, 1:2, 1:2.4;

[0029] Preferably, in the soluble salt solution, the zinc ions are derived from zinc chloride;

[0030] Preferably, in the soluble salt solution, the calcium ions are derived from calcium chloride.

[0031] A second aspect of the present invention relates to a method for preparing another pharmaceutical mixture, comprising the steps of:

[0032] 1) providing a soluble salt solution comprising zinc ions, aluminum ions and manganese ions;

[0033] 2) providing an alkaline solution comprising phosphate ions and hydroxide ions;

[0034] 3) Mixing the soluble salt solution with the alkaline solution to obtain a mixture.

[0035] In some embodiments of the second aspect of the present invention, the method has one or more of the following features A to G:

[0036] A. The molar concentration of zinc ions in the soluble salt solution is 20 to 64 mM, for example, 23 mM, 25 mM, 27 mM, 30 mM, 32 mM, 34 mM, 36 mM, 38 mM, 41 mM, 43 mM, 45 mM, 47 mM, 48 mM, 50 mM, 52 mM, 55 mM, 57 mM, 58 mM, 60 mM, 62 mM;

[0037] B. The molar ratio of zinc ion, aluminum ion and manganese ion in the soluble salt solution is 1:(1-4):(0.5-2.0), preferably 1:(1-2):1, for example 1:2:1, 1:3:0.5;

[0038] C. The molar concentration of phosphate ions in the alkaline solution is 4 to 30 mM, for example, 6 mM, 8 mM, 10 mM, 12 mM, 14 mM, 16 mM, 18 mM, 20 mM, 22 mM, 24 mM, 26 mM, 28 mM;

[0039] D. The molar concentration of hydroxide ions in the alkaline solution is 120 to 170 mM, for example, 130 mM, 135 mM, 140 mM, 145 mM, 150 mM, 155 mM, 160 mM, 165 mM, 170 mM;

[0040] E. In step 3), the volume ratio of the soluble salt solution to the alkaline solution is 1:(0.5-2), for example 1:1, 1:1.5;

[0041] F. In step 3), mixing is performed by adding (e.g., dropping) the alkaline solution into the soluble salt solution;

[0042] G. The method further comprises step 4): sterilizing the mixture to obtain a sterilized product;

[0043] More preferably, the temperature of the sterilization treatment is 100°C to 140°C, such as 110°C, 115°C, 120°C, 125°C, 130°C;

[0044] More preferably, the sterilization treatment time is 0.5 to 3 hours, for example 1 hour, 1.5 hours, 2 hours, 2.5 hours;

[0045] More preferably, the pH value of the sterilized product is adjusted to 6 to 7, for example 6.5.

[0046] The third aspect of the present invention relates to a pharmaceutical mixture prepared by the method described in the first aspect of the present invention.

[0047] In some embodiments of the third aspect of the present invention, the pharmaceutical mixture comprises multiple substances selected from the group consisting of calcium chloride, calcium phosphate, calcium zoledronic acid, calcium risedronate, zinc chloride, zinc phosphate, zinc hydroxide, zinc zoledronic acid and zinc risedronate.

[0048] In some embodiments of the third aspect of the present invention, the pharmaceutical mixture comprises calcium chloride, calcium phosphate, zinc chloride, zinc hydroxide, zinc phosphate, calcium zoledronic acid and zinc zoledronic acid.

[0049] In some embodiments of the third aspect of the present invention, the pharmaceutical mixture comprises zinc chloride, zinc hydroxide, zinc phosphate and zinc zoledronic acid.

[0050] In some embodiments of the third aspect of the present invention, the pharmaceutical mixture comprises zinc chloride, zinc hydroxide, zinc phosphate and risedronate zinc.

[0051] The fourth aspect of the present invention relates to another pharmaceutical mixture prepared by the method described in the second aspect of the present invention.

[0052] In some embodiments of the fourth aspect of the present invention, the pharmaceutical mixture comprises multiple substances selected from the group consisting of ZnCl2, AlCl3, MnCl2, Zn(OH)2, Al(OH)3, Mn(OH)2, AlPO4 and aluminum hydroxyphosphate.

[0053] The fifth aspect of the present invention relates to a pharmaceutical preparation comprising the pharmaceutical mixture described in the third or fourth aspect of the present invention and optional pharmaceutical excipients.

[0054] In some embodiments of the fifth aspect of the present invention, the pharmaceutical preparation of the present invention is a nasal spray preparation or a nasal drop preparation.

[0055] Liquid pharmaceutical preparations can be taken orally or externally, and the liquid pharmaceutical preparations of the present invention include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups, elixirs, sprays and drops. Pharmaceutical excipients can be selected from inert diluents (e.g., water or other solvents), solubilizers and emulsifiers commonly used in the art, such as ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, oil (specifically cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil and sesame oil), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycol, fatty acid esters of sorbitan, and mixtures thereof. In liquid preparations, pharmaceutical excipients can also be selected from adjuvants such as wetting agents, emulsifiers and suspending agents, sweeteners, flavoring agents, colorants, flavoring agents and preservatives. In the suspension preparation, the pharmaceutical excipient may be a suspending agent, for example, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar and tragacanth gum, and mixtures thereof.

[0056] The pharmaceutical preparation of the present invention for rectal or vaginal administration can be a suppository, which can be prepared by mixing the pharmaceutical mixture with one or more suitable non-irritating excipients or carriers (including, for example, cocoa butter, polyethylene glycol, suppository wax or salicylate), and is solid at room temperature and liquid at body temperature, so that it will melt in the rectum or vagina to release the zinc-aluminum complex adjuvant. The preparation suitable for vaginal administration of the present invention also includes vaginal suppositories, tampons, creams, gels, pastes, foams or sprays containing suitable carriers known in the art.

[0057] The pharmaceutical preparations of the present invention suitable for parenteral administration comprise a pharmaceutical mixture and one or more pharmaceutically acceptable sterile isotonic aqueous or non-aqueous carriers in combination therewith, including solutions, dispersions, suspensions or emulsions or sterile powders that can be reconstituted into sterile injectable solutions or dispersions before use, which may contain antioxidants, buffers, solutes that make the preparation isotonic with the blood of the intended recipient, suspending agents or thickening agents.

[0058] Examples of suitable aqueous and non-aqueous carriers that can be used in the pharmaceutical preparations of the present invention include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, etc.) and suitable mixtures thereof, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate. Appropriate fluidity can be maintained, for example, by using coating materials such as lecithin, by maintaining the desired particle size (in the case of dispersants), and by using surfactants.

[0059] The pharmaceutical preparation of the present invention may also contain adjuvants such as wetting agents, emulsifiers and dispersants. It may also be necessary to include isotonic agents such as sugars, sodium chloride, etc. In addition, the extended absorption of injectable drug forms can be achieved by incorporating substances that can delay absorption such as aluminum monostearate and gelatin.

[0060] The dosage of the pharmaceutical preparations of the present invention is determined by techniques well known to those skilled in the pharmaceutical art, taking into account factors such as the age, sex, weight, species and condition of the particular animal or patient, and the route of administration.

[0061] In one embodiment, the pharmaceutical preparation of the present invention further comprises a component selected from the group consisting of surfactants, absorption enhancers, water-absorbing polymers, substances that inhibit enzymatic degradation, alcohols, organic solvents, oils, pH control agents, preservatives, osmotic pressure control agents, propellants, water, and any mixtures thereof.

[0062] The pharmaceutical preparations of the present invention may conveniently be administered by oral, parenteral (subcutaneous, intramuscular, intravenous, dermal, intraperitoneal or intraperitoneal), intrabuccal, intranasal or transdermal routes.

[0063] The sixth aspect of the present invention relates to the use of at least one selected from zinc chloride, ferric chloride, aluminum chloride and their aqueous solutions, and / or a pharmaceutical mixture in the preparation of a medicament for preventing or treating respiratory tract-transmitted viral infections, or

[0064] Use in the preparation of a medicament for preventing or treating a disease or condition caused by a respiratory virus infection;

[0065] Wherein, the pharmaceutical mixture is the pharmaceutical mixture according to the third or fourth aspect of the present invention, or the pharmaceutical mixture is prepared by a method comprising the following steps:

[0066] i) providing a soluble salt solution comprising at least one selected from zinc ions, aluminum ions and calcium ions;

[0067] ii) providing an alkaline solution comprising phosphate ions and hydroxide ions;

[0068] iii) mixing the soluble salt solution with the alkaline solution to obtain a mixture.

[0069] In some embodiments of the sixth aspect of the present invention, in step iii) of the method, the alkaline solution is added to the soluble salt solution for mixing.

[0070] In some embodiments of the sixth aspect of the present invention, in step iii) of the method, the alkaline solution is dropped into the soluble salt solution for mixing.

[0071] In some embodiments of the sixth aspect of the present invention, the use has one or more of the following features (a) to (f):

[0072] (a) a respiratory virus is selected from influenza A virus, influenza B virus, influenza C virus, influenza D virus, respiratory syncytial virus and a novel coronavirus (e.g., SARS-CoV-2), preferably influenza A virus, influenza B virus, respiratory syncytial virus and a novel coronavirus (e.g., SARS-CoV-2);

[0073] (b) diseases or conditions caused by respiratory viruses selected from seasonal influenza, pandemic influenza, zoonotic influenza, rhinitis, pharyngitis, laryngitis, bronchitis, pneumonia, fever, headache, sore throat, fatigue, muscle aches, nausea and vomiting, dry cough, dyspnea, nasal congestion, runny nose and diarrhea;

[0074] Preferably, seasonal influenza is selected from seasonal influenza type A, type B and type C;

[0075] Preferably, the animal-origin influenza is selected from swine influenza and avian influenza;

[0076] (c) the molar concentration of the aqueous zinc chloride solution, the aqueous ferric chloride solution or the aqueous aluminum chloride solution is 0.8 to 20 mM, for example, 1 mM, 2 mM, 4 mM, 6 mM, 8 mM, 10 mM, 12 mM, 14 mM, 15 mM, 17 mM, 18 mM;

[0077] (d) the drug is in the form of nasal spray or nasal drops;

[0078] (e) in step iii), the volume ratio of the soluble salt solution to the alkaline solution is 1:(0.5-2), for example 1:0.8, 1:1, 1:1.5;

[0079] (f) The method further comprises step iv): sterilizing the mixture to obtain a sterilized product.

[0080] In some embodiments of the sixth aspect of the present invention, the sterilization treatment is high-pressure sterilization treatment.

[0081] In some embodiments of the sixth aspect of the present invention, the temperature of the sterilization treatment is 100°C to 140°C, for example, 110°C, 115°C, 120°C, 125°C, 130°C.

[0082] In some embodiments of the sixth aspect of the present invention, the sterilization treatment time is 0.5 to 3 hours, for example, 1 hour, 1.5 hours, 2 hours, or 2.5 hours.

[0083] In some embodiments of the six aspects of the present invention, the pH value of the sterilized material is adjusted to 6-7, for example, 6.5-7.

[0084] In some embodiments of the sixth aspect of the present invention, the pH value is adjusted using an acid solution (eg, hydrochloric acid solution) or an alkaline solution (eg, sodium hydroxide solution).

[0085] In some embodiments of the sixth aspect of the present invention, the sterilized material after the pH value is adjusted is diluted (with water), for example, diluted to 2 to 8 times the volume, or diluted to 2 to 4 times the volume.

[0086] In some embodiments of the sixth aspect of the present invention, the method has any one of the following features a to c:

[0087] a. The soluble salt solution contains zinc ions and aluminum ions;

[0088] Preferably, the molar ratio of zinc ions to aluminum ions in the soluble salt solution is 1:(1-4), such as 1:2, 1:3;

[0089] Preferably, the molar ratio of phosphate ions to hydroxide ions in the alkaline solution is 1:(8-14), for example, 1:8.4, 1:9, 1:9.5, 1:10, 1:10.6, 1:11, 1:11.5, 1:12, 1:12.4, 1:13, 1:13.7;

[0090] Preferably, the molar concentration of zinc ions in the soluble salt solution is 20 to 64 mM, for example 30 mM, 35 mM, 40 mM, 45 mM, 50 mM, 55 mM, 60 mM;

[0091] Preferably, the molar concentration of phosphate ions in the alkaline solution is 4 to 30 mM, for example 6 mM, 10 mM, 14 mM, 16 mM, 18 mM, 20 mM, 23 mM, 26 mM, 28 mM;

[0092] b. The soluble salt solution contains aluminum ions but does not contain calcium ions and zinc ions;

[0093] Preferably, the molar ratio of phosphate ions to hydroxide ions in the alkaline solution is 1:(6-11), for example, 1:6.3, 1:6.7, 1:7.0, 1:7.4, 1:7.8, 1:7.9, 1:8.0, 1:8.3, 1:8.5, 1:9.0, 1:9.4, 1:9.8, 1:10.0, 1:10.4, 1:10.8;

[0094] Preferably, the molar concentration of aluminum ions in the soluble salt solution is 40 to 80 mM, for example 42 mM, 45 mM, 50 mM, 55 mM, 60 mM, 65 mM, 70 mM, 75 mM;

[0095] Preferably, the molar concentration of phosphate ions in the alkaline solution is 3 to 20 mM, for example, 5 mM, 7 mM, 10 mM, 12 mM, 14 mM, 16 mM, 18 mM;

[0096] c. The soluble salt solution contains calcium ions but does not contain zinc ions and aluminum ions;

[0097] Preferably, the molar concentration of calcium ions in the soluble salt solution is 40 to 82 mM, for example 45 mM, 50 mM, 55 mM, 60 mM, 65 mM, 70 mM, 75 mM, 80 mM;

[0098] Preferably, the molar concentration of phosphate ions in the alkaline solution is 40 to 82 mM, for example 45 mM, 50 mM, 55 mM, 60 mM, 65 mM, 70 mM, 75 mM, 80 mM;

[0099] Preferably, the molar ratio of phosphate ions to hydroxide ions in the alkaline solution is (1-7):1, for example, 1.5:1, 1.8:1, 2.0:1, 2.3:1, 2.5:1, 2.8:1, 3:1, 3.4:1, 3.6:1, 4:1, 4.5:1, 4.7:1, 5.0:1, 5.5:1, 6:1, 6.3:1, 6.5:1, 6.7:1.

[0100] The seventh aspect of the present invention relates to a pharmaceutical preparation comprising a pharmaceutical mixture, a recombinant protein RBD of the new coronavirus and optional pharmaceutical excipients; wherein

[0101] The pharmaceutical mixture is the pharmaceutical mixture according to the fourth aspect of the present invention, or the pharmaceutical mixture is prepared by a method comprising the following steps:

[0102] 1) providing a soluble salt solution comprising aluminum ions;

[0103] II) providing an alkaline solution comprising phosphate ions and hydroxide ions;

[0104] III) mixing the soluble salt solution with the alkaline solution to obtain a mixture.

[0105] In some embodiments of the seventh aspect of the present invention, in step II), the molar ratio of phosphate ions to hydroxide ions in the alkaline solution is 1:(6-11), for example, 1:6.3, 1:6.7, 1:7.0, 1:7.4, 1:7.8, 1:7.9, 1:8.0, 1:8.3, 1:8.5, 1:9.0, 1:9.4, 1:9.8, 1:10.0, 1:10.4, and 1:10.8.

[0106] In some embodiments of the seventh aspect of the present invention, in step I), the molar concentration of aluminum ions in the soluble salt solution is 40-80 mM, for example 42 mM, 45 mM, 50 mM, 55 mM, 60 mM, 65 mM, 70 mM, 75 mM.

[0107] In some embodiments of the seventh aspect of the present invention, in step II), the molar concentration of phosphate ions in the alkaline solution is 3-20 mM, for example, 5 mM, 7 mM, 10 mM, 12 mM, 14 mM, 16 mM, 18 mM.

[0108] In some embodiments of the seventh aspect of the present invention, in step III), the volume ratio of the soluble salt solution to the alkaline solution is 1:(0.5-2), for example 1:0.8, 1:1, 1:1.5.

[0109] In some embodiments of the seventh aspect of the present invention, the volume ratio of the medicinal mixture to the SARS-CoV-2 recombinant protein RBD is 1:(0.7-2), for example, 1:0.8, 1:1, or 1:1.5.

[0110] In some embodiments of the seventh aspect of the present invention, the pharmaceutical preparation is a nasal spray preparation or a nasal drop preparation.

[0111] The eighth aspect of the present invention relates to the use of a medicinal mixture in combination with a novel coronavirus recombinant protein RBD in the preparation of a medicament for preventing or treating novel coronavirus (e.g., SARS-CoV-2) infection, or

[0112] Use in the preparation of a medicament for preventing or treating a disease or condition caused by infection with a novel coronavirus (e.g., SARS-CoV-2);

[0113] Wherein, the pharmaceutical mixture is the pharmaceutical mixture described in the seventh aspect of the present invention.

[0114] In some embodiments of the eighth aspect of the present invention, the volume ratio of the medicinal mixture and the SARS-CoV-2 recombinant protein RBD is 1:(0.7-2), for example, 1:0.8, 1:1, or 1:1.5.

[0115] In some embodiments of the seventh or eighth aspects of the present invention, the SARS-CoV-2 recombinant protein RBD is produced by a CHO expression system and then purified.

[0116] In some embodiments of the eighth aspect of the present invention, the disease or condition caused by infection with the new coronavirus is selected from rhinitis, pharyngitis, laryngitis, bronchitis, pneumonia, fever, headache, pharyngitis, fatigue, muscle aches, nausea and vomiting, dry cough, dyspnea, nasal congestion, runny nose and diarrhea.

[0117] Another aspect of the present invention relates to a method for preventing or treating a respiratory viral infection, or a method for preventing or treating a disease or condition caused by a respiratory viral infection, comprising administering to a subject in need thereof an effective amount of at least one selected from zinc chloride, ferric chloride, aluminum chloride and their aqueous solutions, or administering to a subject in need thereof an effective amount of the pharmaceutical mixture described in the sixth aspect of the present invention.

[0118] Another aspect of the present invention relates to at least one selected from zinc chloride, ferric chloride, aluminum chloride and their aqueous solutions or the pharmaceutical mixture described in the sixth aspect of the present invention, which is used for preventing or treating respiratory viral infections, or for preventing or treating diseases or conditions caused by respiratory viral infections.

[0119] In some embodiments of the present invention, the respiratory virus is selected from influenza A virus, influenza B virus, influenza C virus, influenza D virus, respiratory syncytial virus and new coronavirus (such as SARS-CoV-2), preferably influenza A virus, influenza B virus, respiratory syncytial virus and new coronavirus (such as SARS-CoV-2).

[0120] In some embodiments of the present invention, the disease or condition caused by a respiratory virus is selected from seasonal influenza, pandemic influenza, zoonotic influenza, rhinitis, pharyngitis, laryngitis, (bronchitis), pneumonia, fever, headache, sore throat, fatigue, muscle aches, nausea and vomiting, dry cough, dyspnea, nasal congestion, runny nose and diarrhea.

[0121] In some embodiments of the present invention, the molar concentration of the aqueous zinc chloride solution, the aqueous ferric chloride solution or the aqueous aluminum chloride solution is 0.8 to 20 mM, for example, 1 mM, 2 mM, 4 mM, 6 mM, 8 mM, 10 mM, 12 mM, 14 mM, 15 mM, 17 mM, or 18 mM.

[0122] Another aspect of the present invention relates to a method for preventing or treating infection with a novel coronavirus (e.g., SARS-CoV-2), or preventing or treating a disease or condition caused by infection with a novel coronavirus (e.g., SARS-CoV-2), comprising administering to a subject in need thereof the pharmaceutical mixture described in the seventh aspect of the present invention and the novel coronavirus recombinant protein RBD.

[0123] Another aspect of the present invention relates to the combination of the pharmaceutical mixture described in the seventh aspect of the present invention and the new coronavirus recombinant protein RBD, which is used to prevent or treat infection with the new coronavirus (such as SARS-CoV-2), or to prevent or treat diseases or symptoms caused by infection with the new coronavirus (such as SARS-CoV-2).

[0124] In some embodiments of the present invention, the volume ratio of the medicinal mixture to the SARS-CoV-2 recombinant protein RBD is 1:(0.7-2), for example, 1:0.8, 1:1, or 1:1.5.

[0125] In some embodiments of the present invention, the SARS-CoV-2 recombinant protein RBD is produced by a CHO expression system and then purified.

[0126] In some embodiments of the present invention, the disease or condition caused by infection with the new coronavirus is selected from rhinitis, pharyngitis, laryngitis, bronchitis, pneumonia, fever, headache, pharyngitis, fatigue, muscle aches, nausea and vomiting, dry cough, dyspnea, nasal congestion, runny nose and diarrhea.

[0127] In the present invention, the soluble salt solution is an aqueous solution of a soluble salt. In the soluble salt solution, zinc ions, calcium ions, aluminum ions and manganese ions are derived from soluble zinc salts, soluble calcium salts, soluble aluminum salts and soluble manganese salts, such as zinc chloride, calcium chloride, aluminum chloride and manganese chloride, respectively.

[0128] In the present invention, the alkaline solution is an aqueous solution.

[0129] In the present invention, in the alkaline solution, the phosphate ions are derived from soluble phosphates, hydrogen phosphates, dihydrogen phosphates or their crystalline hydrates, such as sodium phosphate, sodium hydrogen phosphate, sodium dihydrogen phosphate.

[0130] In the present invention, in the alkaline solution, the bisphosphonate ions are derived from soluble bisphosphonates or their crystalline hydrates, such as sodium bisphosphonate and potassium bisphosphonate.

[0131] In the present invention, in the alkaline solution, the hydroxide ions are derived from hydroxides, such as sodium hydroxide and potassium hydroxide.

[0132] In the present invention, the structural formula of the zoledronic acid ion is

[0133] In the present invention, the structural formula of the risedronate ion is

[0134] In the present invention, calcium zoledronic acid is a salt formed by calcium ions and zoledronic acid ions.

[0135] In the present invention, risedronate calcium is a salt formed by calcium ions and risedronate ions.

[0136] In the present invention, zinc zoledronic acid is a salt formed by zinc ion and zoledronic acid anion.

[0137] In the present invention, risedronate zinc is a salt formed by zinc ion and risedronate ion.

[0138] The present invention achieves at least one of the following beneficial effects:

[0139] 1. The pharmaceutical mixture prepared by the method of the present invention, or zinc chloride, ferric chloride, aluminum chloride or their aqueous solutions can prevent or treat respiratory tract-transmitted viral infections, or prevent or treat diseases or symptoms caused by respiratory tract-transmitted viral infections.

[0140] 2. The nasal spray preparation or nasal drop preparation prepared by the method of the present invention can prevent or treat respiratory tract-transmitted viral infections, or prevent or treat diseases or symptoms caused by respiratory tract-transmitted viral infections. BRIEF DESCRIPTION OF THE DRAWINGS

[0141] In order to make the content of the present invention more clearly understood, the present invention is further described in detail below according to specific embodiments of the present invention in conjunction with the accompanying drawings, wherein

[0142] Figure 1 Schematic diagram of the co-precipitation method;

[0143] Figure 2A This is the weight change rate curve of mice in the nasal spray immune enhancement preparation FH-001 nasal drops group after the virus challenge;

[0144] Figure 2B This is the survival rate curve of mice in the group of nasal administration of the nasal spray immune enhancement preparation FH-001 after virus challenge;

[0145] Figure 2C This is the weight change rate curve of mice in the intramuscular administration group of the nasal spray immune enhancement preparation FH-001 after the virus challenge;

[0146] Figure 2D This is the survival rate curve of mice in the intramuscular administration group of the nasal spray immune enhancement preparation FH-001 after virus challenge;

[0147] Figure 3A This is the weight change rate curve of mice in the nasal spray immune enhancement preparation FH-001 nasal drops administration group and intraperitoneal administration group after virus challenge;

[0148] Figure 3B This is the survival rate curve of mice in the nasal spray immune enhancement preparation FH-001 nasal drops administration group and intraperitoneal administration group after virus challenge;

[0149] Figure 3C This is the weight change rate curve of mice in the group of intragastric administration of the nasal spray immune enhancement preparation FH-001 after the virus challenge;

[0150] Figure 3D This is the survival rate curve of mice in the group of intragastric administration of the nasal spray immune enhancement preparation FH-001 after virus challenge;

[0151] Figure 4A This is the weight change rate curve of mice in the group of nasal administration of 100 μL of the nasal spray immune enhancement preparation FH-001 after the virus challenge;

[0152] Figure 4B This is the survival rate curve of mice in the group of nasal administration of 100 μL of the nasal spray immune enhancement preparation FH-001 after virus challenge;

[0153] Figure 4C This is the weight change rate curve of mice in the nasal spray immune enhancement preparation FH-001 30μL and 60μL groups after challenge;

[0154] Figure 4D This is the survival rate curve of mice in the nasal spray immune enhancement preparation FH-001 nasal drops 30μL and 60μL groups after virus challenge;

[0155] Figure 5A This is the weight change rate curve of mice in the nasal spray immune enhancement preparation FH-001 diluted 10 times and 150 times after the virus was challenged;

[0156] Figure 5B This is the survival rate curve of mice in the nasal spray immune enhancement preparation FH-001 diluted 10 times and 150 times after the virus was challenged;

[0157] Fig. 6A The weight change rate curve of mice in the challenge group 1 day after administration of the nasal spray immune enhancement preparation FH-001 and 4 days after administration;

[0158] Figure 6B The survival curves of mice in the challenge group 1 day after administration of the nasal spray immune enhancement preparation FH-001 and 4 days after administration of the challenge group;

[0159] Figure 6C The weight change rate curve of mice in the challenge group 15 days after administration of the nasal spray immune enhancement preparation FH-001 diluted 10 times and 18 days after administration;

[0160] Fig.6DThe survival curves of mice in the challenge group 15 days after administration of the nasal spray immune enhancement preparation FH-001 diluted 10 times and 18 days after administration;

[0161] Fig. 7A This is the weight change rate curve of mice in the nasal spray immune enhancement preparation Al-001 nasal drops group after the virus challenge;

[0162] Figure 7B This is the survival rate curve of mice in the group of nasal administration of Al-001, a nasal spray immune enhancement preparation, after the virus challenge;

[0163] Fig. 8A This is the weight change rate curve of mice in the nasal spray immune enhancement preparation CaP nasal drops group after the virus challenge;

[0164] Figure 8B This is the survival rate curve of mice in the group of nasal spray immune enhancement preparation CaP nasal drops after challenge;

[0165] Fig.9A This is the weight change rate curve of mice in the group of nasal administration of bisphosphate zinc nasal drops of nasal spray immune enhancement preparation after the virus attack;

[0166] Fig. 9B This is the survival rate curve of mice in the group of nasal administration of bisphosphate zinc nasal drops of nasal spray immune enhancement preparation after virus attack;

[0167] Fig. 10A This is the weight change rate curve of mice in the group of nasal administration of organic bisphosphonate zinc-1, a nasal spray immune enhancement preparation, after the virus was challenged;

[0168] Fig. 10B This is the survival rate curve of mice in the group of nasal administration of organic bisphosphonate zinc-1, a nasal spray immune enhancement preparation, after the virus was challenged;

[0169] Fig. 10C This is the weight change rate curve of mice in the group of nasal administration of organic bisphosphonate zinc-2, a nasal spray immune enhancement preparation, after the virus challenge;

[0170] Fig. 10D This is the survival rate curve of mice in the group of nasal administration of zinc bisphosphonate-2, a nasal spray immune enhancement preparation, after the virus was challenged;

[0171] Fig.11A This is the weight change rate curve of mice in the group of nasal spray immune enhancement preparation containing zinc solution after the virus challenge;

[0172] Fig. 11B This is the survival rate curve of mice in the group of nasal spray immune enhancement preparation containing zinc solution after virus challenge;

[0173] Fig. 11C This is the weight change rate curve of mice in the group of nasal spray immune enhancement preparation containing aluminum solution after challenge;

[0174] Fig.11D This is the survival rate curve of mice in the group of nasal spray immune enhancement preparations containing aluminum solution after challenge;

[0175] Fig.11E This is the weight change rate curve of mice in the group of nasal administration of iron solution containing nasal spray immune enhancement preparation after the virus challenge;

[0176] Fig.11F This is the survival rate curve of mice in the group of nasal administration of iron solution containing nasal spray immune enhancement preparation after virus challenge;

[0177] Fig.12 This is the curve of the change of virus titer in the lungs of mice in the group of nasal administration of the nasal spray immune enhancement preparation FH-001 after the virus was challenged;

[0178] Fig.13A HE staining of the lungs of mice in the group receiving nasal administration of the nasal spray immune enhancement preparation FH-001 after challenge;

[0179] Fig. 13B HE staining of the lungs of mice in the negative control group after challenge;

[0180] Fig. 13C HE staining of the lungs of mice in the blank group;

[0181] Fig.14A This is the weight change rate curve of mice in the nasal spray immune enhancement preparation FH-001 nasal drops group after challenge with influenza virus (A / California / 04 / 2009);

[0182] Fig. 14B This is the survival rate curve of mice in the nasal spray immune enhancement preparation FH-001 nasal drops group after challenge with influenza virus (A / California / 04 / 2009);

[0183] Fig.15A This is the weight change rate curve of mice in the nasal spray immune enhancement preparation FH-001 nasal drops group after challenge with respiratory syncytial virus RSV;

[0184] Fig. 15B The virus titer level of the nose and lung tissues of mice in the group of intranasal administration of the nasal spray immune enhancement preparation FH-001 on the 6th day after the virus (respiratory syncytial virus RSV) was challenged;

[0185] Fig.16A This is a transmission electron micrograph of the nasal spray immune enhancement preparation Al-001;

[0186] Fig. 16B This is a transmission electron micrograph of the nasal spray immune enhancement preparation FH-001;

[0187] Fig. 16CThis is a transmission electron micrograph of CaP, a nasal spray immune enhancement preparation;

[0188] Fig.16D This is a transmission electron micrograph of the nasal spray immune enhancement preparation calcium zinc bisphosphate;

[0189] Fig.17A The appearance of the nasal spray immune enhancement preparations FH-001, FH-001-Mn, FH-001-Mn + 5% sucrose and FH-001-Mn + 5% trehalose;

[0190] Fig. 17B The appearance of the nasal spray immune enhancement preparations FH-001, FH-001-Mn, FH-001-Mn + 5% sucrose and FH-001-Mn + 5% trehalose after sedimentation for 12 hours;

[0191] Fig.18A This is the curve of the weight change rate of mice in the group that were first pretreated with CLL and then nasally sprayed with the immune enhancement agent FH-001 after the virus was challenged;

[0192] Fig.18B This is the survival rate curve of mice in the group that were first pretreated with CLL and then nasally sprayed with the immune enhancement agent FH-001 after virus attack;

[0193] Fig.19 This is the change over time of antibody titers in mice in the group administered with the nasal spray immune enhancing preparation FH-001-Mn combined with the recombinant protein RBD of the new coronavirus, and the group administered with the nasal spray immune enhancing preparation Al-001 combined with the recombinant protein RBD of the new coronavirus. DETAILED DESCRIPTION

[0194] The embodiments of the present invention will be described clearly and completely below in conjunction with the examples. Obviously, the described examples are only some examples of the present invention, rather than all examples. The following description of at least one exemplary embodiment is actually only illustrative and is by no means intended to limit the present invention and its application or use. Based on the examples in the present invention, all other examples obtained by ordinary technicians in the field without creative work are within the scope of protection of the present invention.

[0195] Preparation Example 1 Preparation of Nasal Spray Immunoenhancing Preparation FH-001

[0196] Prepare an aqueous solution containing 46.67 mM ZnCl2 and 124.44 mM AlCl3 (Solution A) and 18.67 mM PO4 3- and 200mM sodium hydroxide aqueous solution (B solution), the volumes of A and B solutions are equal. A and B solutions are co-precipitated, that is, Figure 1As shown, solution B is added dropwise to solution A until it is completely added; then it is sterilized at 121°C for 60 min and the pH is adjusted to between 6.5 and 7.0 to obtain nasal spray immune enhancement preparation 4×FH-001 (depending on different experimental designs, it can be diluted with sterile saline), which mainly contains ZnCl2, AlCl3, Zn(OH)2, Al(OH)3, AlPO4 and aluminum phosphate hydroxide [Al x (OH) y (PO4) z ]wait.

[0197] Preparation Example 2 Preparation of Nasal Spray Immunoenhancing Preparation Al-001

[0198] Formulated to contain 62.22 mM Al 3+ AlCl3 aqueous solution (Solution A) and 9.33mM PO4 3- and 75mM sodium hydroxide aqueous solution (B solution), the volumes of A and B solutions are equal. A and B solutions are co-precipitated, that is, Figure 1 As shown, solution B is added dropwise to solution A until it is completely added; then it is sterilized at 121°C for 60 minutes and the pH is adjusted to between 6.5 and 7.0 to obtain the nasal spray immune enhancement preparation 2×Al-001 (depending on different experimental designs, it can be diluted with sterile saline), which mainly contains aluminum hydroxide phosphate [Al x (OH) y (PO4) z ], AlPO4, Al(OH)3, AlCl3, etc.

[0199] Preparation Example 3 Preparation of Nasal Spray Immunoenhancing Preparation CaP

[0200] Formulated with 62.22 mM Ca 2+ Aqueous solution of CaCl2 (Solution A) and 62.22 mM PO4 3- and 20.00 mM sodium hydroxide aqueous solution (B solution), the volumes of A and B solutions are equal. A and B solutions are co-precipitated, that is, Figure 1 As shown, solution B is added dropwise into solution A until it is completely added; then, it is sterilized at 121°C for 60 minutes under high pressure, and the pH is adjusted to be between 6.5 and 7.0 to prepare a nasal spray immune enhancement preparation CaP, which mainly contains CaCl2, Ca3(PO4)2, etc.

[0201] Preparation Example 4 Preparation of Nasal Spray Immunoenhancing Preparation FH-001-Mn

[0202] Prepare an aqueous solution (Solution A) containing 49.78 mM ZnCl2, 74.66 mM AlCl3 and 49.78 mM MnCl2 and a solution containing 18.67 mM PO4 3- and 145 mM sodium hydroxide aqueous solution (B solution), with equal volumes of A and B solutions. A and B solutions are co-precipitated, i.e. Figure 1 As shown, solution B is added dropwise into solution A until it is completely added; then, the solution is sterilized at 121° C. for 60 min and the pH is adjusted to be between 6.5 and 7.0 to obtain the nasal spray immune enhancement preparation 4×FH-001-Mn.

[0203] Preparation Example 5 Preparation of nasal spray immunopotentiating preparation calcium zinc bisphosphate

[0204] Prepare an aqueous solution containing 46.67 mM ZnCl2, 46.67 mM CaCl2 (Solution A) and 42 mM PO4 3- , 7.8 mM organic bisphosphonate ion (zoledronic acid ion) and 40 mM sodium hydroxide aqueous solution (B solution), A and B solutions are equal in volume. A and B solutions are coprecipitated to prepare calcium zinc bisphosphonate, that is, Figure 1 As shown, solution B is added dropwise into solution A until it is completely added; then, the solution is sterilized at 121° C. for 60 min under high pressure, and the pH is adjusted to be between 6.5 and 7.0 to prepare a nasal spray immune enhancement preparation of calcium zinc bisphosphonate, which mainly contains CaCl2, Ca3(PO4)2, ZnCl2, Zn(OH)2, Zn3(PO4)2, zoledronic acid calcium and zoledronic acid zinc.

[0205] Preparation Example 6 Preparation of Nasal Spray Immunoenhancing Preparations: Organobisphosphonate Zinc-1 and Organobisphosphonate Zinc-2

[0206] (1) Preparation of organic bisphosphonate zinc-1: Prepare a solution containing 62.22 mM Zn 2+ Aqueous ZnCl2 solution (Solution A) and 42 mM PO4 3- , 7.8 mM zoledronic acid ion and 66 mM sodium hydroxide aqueous solution (B solution), A and B solutions are equal in volume. A and B solutions are coprecipitated to prepare organic bisphosphonate zinc-1, that is, Figure 1 As shown, solution B is added drop by drop into solution A until it is completely added; then it is sterilized at 121°C for 60 minutes under high pressure, and the pH is adjusted to ensure that it is between 6.5 and 7.0 to prepare a nasal spray immune enhancement preparation organic bisphosphonate zinc-1, which mainly contains ZnCl2, Zn(OH)2, Zn3(PO4)2 and zinc zoledronic acid.

[0207] (2) Preparation of organic bisphosphonate zinc-2: Prepare a solution containing 62.22 mM Zn 2+ Aqueous ZnCl2 solution (Solution A) and 42 mM PO43- , 7.8 mM risedronate ion and 60 mM sodium hydroxide aqueous solution (B solution), A and B solutions are equal in volume. A and B solutions are coprecipitated to prepare organic bisphosphonate zinc-2, that is, Figure 1 As shown, solution B is added drop by drop into solution A until it is completely added; then, the solution is sterilized at 121°C for 60 min under high pressure, and the pH is adjusted to be between 6.5 and 7.0 to prepare a nasal spray immune enhancement preparation, organic bisphosphonate zinc-2, which mainly contains ZnCl2, Zn(OH)2, Zn3(PO4)2 and risedronate zinc.

[0208] Preparation Example 7 Preparation of nasal spray immune enhancement preparation containing zinc solution, aluminum solution and iron solution

[0209] Deionized water was used to prepare 2+ 、Al 3+ , Fe 3+ The ZnCl2, AlCl3 and FeCl3 aqueous solutions are then filtered and sterilized. The filtered ZnCl2 aqueous solution is diluted 100 times with sterile saline to prepare a zinc solution for the nasal spray immune enhancement preparation; the filtered AlCl3 and FeCl3 solutions are diluted 10 times with sterile saline to prepare aluminum and iron solutions for the nasal spray immune enhancement preparation.

[0210] The experimental materials used in the following examples are as follows:

[0211] The nasal spray immune enhancement preparation FH-001 was prepared and stored in this laboratory;

[0212] B / Florida / 4 / 2006 Yamagata Lineage is deposited in this laboratory;

[0213] SPF Balb / c mice were purchased from Shanghai Slake Experimental Animal Co., Ltd. and Lingchang Biotechnology Co., Ltd., and SPF female Balb / c mice aged 6-8 weeks were selected for the experiment;

[0214] Physiological saline was prepared with sodium chloride (purchased from Xilong Chemical Company) and stored after filtration;

[0215] Isoflurane was purchased from Shenzhen Reward Life Science Co., Ltd.

[0216] Example 1 Nasal spray immune enhancement preparation FH-001 was administered twice by nasal drops for influenza B virus B / Florida / 4 / 2006 Yamagata The protective effect of Lineage infection on mice

[0217] 1) Drug dilution: Use normal saline to dilute the nasal spray immune enhancement preparation FH-001 by 2 times.

[0218] 2) Administration to Balb / c mice: The mice were randomly divided into 4 groups, namely, the nasal spray immune enhancement preparation FH-001 nasal drops administration group, the saline nasal drops administration group, the nasal spray immune enhancement preparation FH-001 intramuscular administration group, and the saline intramuscular administration group. The nasal drops administration group was anesthetized with isoflurane before administration, and the intramuscular administration group was administered with a syringe, with a volume of 100 μL per mouse, and two injections were used for administration on days 0 and 7.

[0219] 3) Balb / c mice challenge: The mice were challenged with isoflurane anesthesia on the 14th day after the second administration. The mice were anesthetized with 50 μL of 25 LD intranasal drops. 50 The body weight of mice was monitored starting from the 14th day after the second administration (i.e., the day of challenge) and continued for 14 days.

[0220] The results are as follows Figures 2A to 2D As shown in the figure, mice were judged dead when their body weight dropped to 75% of the weight before the challenge on the 14th day. Figure 2A , 2B They are the body weight change curve and survival curve of the nasal drops group. The maximum weight loss of the nasal spray immune enhancement preparation FH-001 nasal drops group did not exceed 10%, and the survival rate was 100%, while all the mice in the normal saline nasal drops group died on the 9th day after the infection. Figure 2C , 2D The weight change curve and survival curve of the intramuscular administration group are shown respectively. All mice died on the 9th day after the virus challenge. The above results prove that the intranasal administration of the nasal spray immune enhancement preparation FH-001 twice can 100% protect mice from the lethal dose infection of B / Florida / 4 / 2006 Yamagata Lineage virus, while the intramuscular administration has no protective effect.

[0221] Example 2 Nasal spray immune enhancement preparation FH-001 nasal drops once for influenza B virus B / Florida / 4 / 2006 Yamagata The protective effect of Lineage infection on mice

[0222] The mice were randomly divided into two groups: the nasal spray immune enhancement preparation FH-001 nasal drops / intraperitoneal / oral administration group and the saline nasal drops / intraperitoneal / oral administration group. Each mouse was administered with a volume of 100 μL, and the administration was carried out by one injection on day 0. The mice were challenged with the poison 7 days after administration. After the mice were anesthetized with isoflurane, 50 μL of nasal drops with a concentration of 25 LD 50 The B / Florida / 4 / 2006 Yamagata Lineage virus was used. The weight changes of mice were monitored starting from the 7th day after administration and continued for 14 days (the mice were considered dead when their weight dropped to 75% of the weight before the challenge on the 7th day).

[0223] The results are as follows Figures 3A to 3D As shown, Figure 3A and Figure 3B The following are the body weight change curves and survival curves of the nasal / intraperitoneal administration groups, respectively. The maximum body weight decrease in the nasal administration group of the nasal spray immune enhancement preparation FH-001 did not exceed 10%, and the survival rate was 100%, while all the mice in the normal saline nasal and intraperitoneal administration groups died within 8 days after the infection.

[0224] Figure 3C and Figure 3D They are the body weight change curve and survival curve of the oral administration group. All mice in the oral administration group of nasal spray immune enhancement preparation FH-001 lost weight within 7 days after the virus challenge, and 3 mice died; all mice in the oral administration group of normal saline lost weight within 7 days after the virus challenge, and 4 mice died; the weight loss curves of nasal spray immune enhancement preparation and normal saline oral administration were basically consistent.

[0225] The above results prove that a single nasal administration of the nasal spray immune enhancement preparation FH-001 can 100% protect mice from infection with a lethal dose of B / Florida / 4 / 2006 Yamagata Lineage virus, while intraperitoneal and oral administration have almost no protective effect.

[0226] Example 3 Effect of different volumes of nasal spray immune enhancement preparation FH-001 on influenza B virus B / Florida / 4 / 2006 Yamagata The protective effect of Lineage infection on mice

[0227] The mice were randomly divided into three groups. Each mouse in the first group was given 30 μL of nasal spray immune enhancement preparation FH-001 by nasal drops, each mouse in the second group was given 60 μL of nasal spray immune enhancement preparation FH-001 by nasal drops, and each mouse in the third group was given 100 μL of nasal spray immune enhancement preparation FH-001 by nasal drops. The administration was carried out by 1 injection on day 0. The mice were challenged with poison 7 days after administration. After the mice were anesthetized with isoflurane, 50 μL of 25LD 50 The B / Florida / 4 / 2006 Yamagata Lineage virus was used. The weight changes of mice were monitored starting from the 7th day after administration and continued for 14 days.

[0228] The results are as follows Figures 4A to 4D As shown, the maximum weight loss of mice in the three groups did not exceed 10%, while all the mice in the negative control group died within 7 days after the infection, indicating that different volumes (30 μL, 60 μL, 100 μL) of nasal spray immune enhancement preparations can 100% protect mice from infection with a lethal dose of B / Florida / 4 / 2006 Yamagata Lineage virus.

[0229] Example 4 Effect of different dilution multiples (i.e. different concentrations) of nasal spray immune enhancement preparation FH-001 on acetylcholine Influenza virus type B / Florida / 4 / 2006 Yamagata The protective effect of Lineage infection on mice

[0230] The mice were randomly divided into two groups. One group was administered with 10-fold diluted nasal spray immune enhancement preparation FH-001 by intranasal drops, and the other group was administered with 150-fold diluted nasal spray immune enhancement preparation FH-001 by intranasal drops. The volume of administration was 100 μL per mouse, and the administration was carried out by one injection on day 0. The mice were challenged with poison 7 days after administration. After the mice were anesthetized with isoflurane, 50 μL of 25LD 50 The B / Florida / 4 / 2006 Yamagata Lineage virus was used. The weight changes of mice were monitored starting from the 7th day after administration and continued for 14 days (the mice were considered dead when their weight dropped to 75% of the weight before the challenge on the 7th day).

[0231] The results are as follows Figures 5A to 5B As shown, the maximum weight loss of mice in the 10-fold dilution group did not exceed 10%, and the mice were 100% protected; after the nasal spray immune enhancement preparation was diluted 150 times, it was still 100% able to protect mice from infection with a lethal dose of B / Florida / 4 / 2006 Yamagata Lineage virus.

[0232] Example 5 Nasal spray immune enhancement preparation FH-001 nasal drops for influenza B virus B / Florida / 4 / 2006 Yamagata Duration of protective effect in lineage-infected mice

[0233] The mice were randomly divided into four groups. All mice were given one injection on day 0. The first group was challenged with the nasal spray immune enhancement preparation FH-001 one day after administration, the second group was challenged with the nasal spray immune enhancement preparation FH-001 four days after administration, the third group was challenged with the nasal spray immune enhancement preparation FH-001 diluted 10 times 15 days after administration, and the fourth group was challenged with the nasal spray immune enhancement preparation FH-001 diluted 10 times 18 days after administration. The challenge method was that after the mice were anesthetized with isoflurane, 50 μL of 25LD 50 The body weight of all mice was monitored from the day of infection and continued for 14 days.

[0234] The results are as follows Figures 6A to 6D As shown, Fig. 6A and 6BThe body weight change curve and survival curve of mice challenged with poison 1 day or 4 days after nasal administration of nasal spray immune enhancement preparation were presented. The group challenged with poison 4 days after administration could provide 100% protection to the mice, and one mouse died in the group challenged with poison 6 days after administration, which could provide 80% protection to the mice. Figure 6C and 6D The weight change curve and survival curve of mice challenged with poison 15 days after administration of 10-fold diluted nasal spray immune enhancement preparation and 18 days after administration of poison. The maximum weight loss of mice in the challenge group 15 days after administration did not exceed 10%, and the mice were 100% protected from the lethal dose of poison; all mice in the challenge group 18 days after administration died within 10 days after the challenge; all negative control mice died after the challenge. This shows that the nasal spray immune enhancement preparation can prevent and protect against the lethal dose infection of B / Florida / 4 / 2006 Yamagata Lineage 1 to 15 days after the nasal administration.

[0235] Example 6 Nasal spray immune enhancement preparation Al-001 nasal drops for influenza B virus B / Florida / 4 / 2006 Yamagata The protective effect of Lineage infection on mice

[0236] The mice were randomly divided into two groups, namely the nasal spray immune enhancement preparation Al-001 nasal drops group and the normal saline nasal drops group; among them, the nasal drops group was anesthetized with isoflurane in advance and then administrated, with a volume of 100 μL per mouse, and 2 injections were used, with administration on the 0th day and the 7th day.

[0237] On the 14th day after the second administration, the mice were challenged with isoflurane anesthesia and nasal drops of 50 μL at a concentration of 25 LD 50 The body weight of mice was monitored starting from the 14th day after the second administration (i.e., the day of challenge) and continued for 14 days.

[0238] The results are as follows Fig. 7A and 7B As shown, the survival rate of mice was able to reach 80% on the 14th day after the second administration, indicating that the nasal spray immune enhancement preparation Al-001 has a certain preventive and protective effect against the lethal dose of B / Florida / 4 / 2006 Yamagata Lineage.

[0239] Example 7 Nasal spray immune enhancement preparation CaP nasal drops for influenza B virus B / Florida / 4 / 2006 The protective effect of Yamagata Lineage infection on mice

[0240] The mice were randomly divided into two groups, namely the nasal spray immune enhancement preparation CaP nasal drops group and the normal saline nasal drops group; among them, the nasal drops group was anesthetized with isoflurane in advance and then administrated, with a volume of 100 μL per mouse, and 1 injection was used for administration on day 0.

[0241] The mice were challenged with isoflurane anesthesia and nasal dripped with 50 μL of 25 LD 50 The B / Florida / 4 / 2006 Yamagata Lineage virus was used to detect the changes in body weight of mice starting from day 7 after administration and continuing for 14 days.

[0242] The results are as follows Fig. 8A and 8B As shown, mice were given the drug once and challenged with the virus 7 days after administration, and the survival rate of the mice could reach 80%, indicating that the nasal spray immune enhancement preparation CaP has a certain preventive and protective effect against the lethal dose of B / Florida / 4 / 2006 Yamagata Lineage.

[0243] Example 8 Nasal Spray Immunoenhancing Preparation Bisphosphonic Acid Zinc Nasal Drops for Influenza B Virus B / Florida / 4 / 2006 Yamagata The protective effect of Lineage infection on mice

[0244] The mice were randomly divided into two groups, namely the nasal spray immune enhancement preparation calcium zinc bisphosphate nasal drops group and the normal saline nasal drops group; among them, the nasal drops group was anesthetized with isoflurane in advance and then administrated, with a volume of 100 μL per mouse, and 1 injection was used for administration on day 0.

[0245] The mice were challenged with isoflurane anesthesia and nasal dripped with 50 μL of 25 LD 50 The B / Florida / 4 / 2006 Yamagata Lineage virus was used to detect the changes in body weight of mice starting from day 7 after administration and continuing for 14 days.

[0246] The results are as follows Fig.9A and Fig. 9B As shown, Fig.9A The body weight change curve showed that the body weight of the calcium bisphosphate zinc group remained basically unchanged after the virus attack. Fig. 9B The survival curve showed that the survival rate of the bis calcium phosphate zinc administration group was 100%, indicating that the nasal spray immune enhancement preparation bis calcium phosphate zinc has a better preventive and protective effect against the lethal dose of B / Florida / 4 / 2006 Yamagata Lineage.

[0247] Example 9 Nasal spray immune enhancement preparation Organic bisphosphonate zinc-1 and organic bisphosphonate zinc-2 nasal drops for type 2 Influenza virus B / Florida / 4 / 2006 Yamagata The protective effect of Lineage infection on mice

[0248] The mice were randomly divided into three groups, namely, the nasal spray immune enhancement preparation organic bisphosphonate zinc-1 nasal drops group, the nasal spray immune enhancement preparation organic bisphosphonate zinc-2 nasal drops group and the normal saline nasal drops group; among them, the nasal drops group was anesthetized with isoflurane in advance and then dosed, with a dosing volume of 100 μL per mouse, administered by 1 injection, and the drug was given on day 0.

[0249] The mice were challenged with isoflurane anesthesia and nasal dripped with 50 μL of 25 LD 50 The B / Florida / 4 / 2006 Yamagata Lineage virus was used to detect the changes in body weight of mice starting from day 7 after administration and continuing for 14 days.

[0250] The results are as follows FIG. 10A to FIG. 10D As shown, Fig. 10A and Fig. 10C The body weight change curves showed that the body weights of the two organobisphosphonate zinc administration groups remained basically unchanged after the challenge. Fig. 10B and Fig. 10D The survival curves showed that the survival rates of the two organobisphosphonate zinc administration groups were both 100%, indicating that the two nasal spray immune enhancement preparations organobisphosphonate zinc had a better preventive and protective effect against the lethal dose of B / Florida / 4 / 2006 Yamagata Lineage.

[0251] Example 10 Nasal spray immune enhancement preparation containing zinc solution, aluminum solution and iron solution for nasal administration of type B Influenza virus B / Florida / 4 / 2006 Yamagata The protective effect of Lineage infection on mice

[0252] The mice were randomly divided into four groups, namely, the nasal drops group of zinc solution containing nasal spray immune enhancement preparation, the nasal drops group of aluminum solution containing nasal spray immune enhancement preparation, the nasal drops group of iron solution containing nasal spray immune enhancement preparation and the nasal drops group of normal saline; among them, the nasal drops group was anesthetized with isoflurane in advance and then dosed, with a dosing volume of 100 μL for each mouse, and 1 injection was used on day 0.

[0253] The mice were challenged with isoflurane anesthesia and nasal dripped with 50 μL of 25 LD 50 The B / Florida / 4 / 2006 Yamagata Lineage virus was used to detect the changes in body weight of mice starting from day 7 after administration and continuing for 14 days.

[0254] The results are as follows FIG. 11A to FIG. 11F As shown, Fig.11A , Fig. 11C , Fig.11EThe body weight change curves showed that the body weights of the three solution administration groups remained basically unchanged after the challenge. Fig. 11B , Fig.11D , Fig.11F The survival curves showed that the survival rates of the groups treated with the three solutions were all 100%, indicating that the nasal spray immune enhancement preparations containing zinc solution, aluminum solution and iron solution had a better preventive and protective effect against the lethal dose of B / Florida / 4 / 2006 Yamagata Lineage.

[0255] Example 11 Influenza B virus B / Florida / 4 / 2006 Yamagata Lung disease in mice after Lineage challenge Virus titer test results

[0256] The mice were randomly divided into two groups, namely the nasal spray immune enhancement preparation FH-001 nasal drops group and the normal saline nasal drops group; among them, the nasal drops group was anesthetized with isoflurane in advance and then administrated, with a volume of 100 μL per mouse, and 1 injection was used on day 0.

[0257] The mice were challenged with isoflurane anesthesia and nasal dripped with 50 μL of 25 LD 50 B / Florida / 4 / 2006 Yamagata Lineage virus. 0.5 days, 2 days, 4 days, and 6 days after infection, the lung tissues of mice were ground and centrifuged to obtain the supernatant, which was diluted 100 times to detect the virus titer.

[0258] The results are as follows Fig.12 As shown, on the 6th day after the challenge, the virus titer in the lung supernatant of the FH-001 administration group was below the detection line, while the virus titer in the negative control group was greater than 10000 TCID 50 / mL, this result proved that the nasal spray immune enhancement preparation FH-001 has a good preventive and protective effect against influenza B virus B / Florida / 4 / 2006 Yamagata Lineage at the virus titer level.

[0259] Example 12 Influenza B virus B / Florida / 4 / 2006 Yamagata HE in mice lungs after Lineage challenge Staining results

[0260] The mice were randomly divided into three groups, namely the nasal spray immune enhancement preparation FH-001 nasal drops group, the saline nasal drops group and the blank group. Among them, the nasal drops group was anesthetized with isoflurane in advance, and then the drug was administered. The volume of each mouse was 100μL, and the drug was administered by one injection. The drug was administered on the 0th day; the mice were challenged with the poison on the 7th day after the drug administration. After the mice were anesthetized with isoflurane, 50μL of nasal drops with a concentration of 25LD 50On the 12th day after infection, lung tissues of mice in different groups were collected for HE staining.

[0261] The results are as follows Figures 13A to 13C As shown, the lung tissue morphology of the FH-001-treated group was basically the same as that of the blank mice, while the lung tissue of the negative control group showed bronchial wall thickening and lung infiltration. This result demonstrated from the histological level that the nasal spray immune enhancement preparation FH-001 has a good preventive and protective effect against influenza B virus B / Florida / 4 / 2006 Yamagata Lineage.

[0262] Example 13 Nasal spray immune enhancement preparation FH-001 nasal drops for influenza A virus A / California / 04 / 2009 Preventive and protective effects of lethal mouse challenge model

[0263] The mice were randomly divided into two groups, namely the nasal spray immune enhancement preparation FH-001 nasal drops group and the normal saline nasal drops group; among them, the nasal drops group was anesthetized with isoflurane in advance and then administrated, with a dosing volume of 100 μL per mouse, and 1 injection was used for administration on day 0.

[0264] The mice were challenged with isoflurane anesthesia and nasally dripped 50 μL of 25×LD 50 The weight changes of mice were monitored starting from the 7th day after administration (i.e., the day of challenge) and continued for 14 days.

[0265] The results are as follows Figures 14A to 14B As shown in the figure, the body weight of mice in the FH-001 administration group decreased by up to 20% after the virus challenge, while the weight loss trend of the negative control group after the virus challenge was more obvious, with the maximum decrease exceeding 25%. This result proves from the perspective of mouse weight changes that FH-001 has a good preventive and protective effect on the lethal virus challenge mouse model of influenza A virus A / California / 04 / 2009. Fig. 14B The data show the survival rate curve of mice after infection. The results show that the survival rate of mice in the FH-001 nasal administration group was 80% 14 days after infection, while all mice in the negative control group died after infection, with no mice surviving. This result proves the good preventive and protective effect of FH-001 against RSV virus from the perspective of influenza A virus.

[0266] Example 14 Effect of Nasal Spray Immunopotentiating Preparation FH-001 on Mice Infected with Respiratory Syncytial Virus (RSV) by Nasal Drops (Non-lethal model) preventive protective effect

[0267] The mice were randomly divided into two groups, namely the nasal spray immune enhancement preparation FH-001 nasal drops group and the normal saline nasal drops group; among them, the nasal drops group was anesthetized with isoflurane in advance and then administrated, with a dosing volume of 100 μL per mouse, and 1 injection was used for administration on day 0.

[0268] The mice were challenged with isoflurane anesthesia and nasally dripped 50 μL of 25×LD 50 The weight of the mice was monitored starting from the 7th day after administration (i.e. the day of challenge) and continued for 14 days. In addition, the virus titer was detected by taking the nasal and lung tissues of the mice on the 6th day after challenge.

[0269] The results are as follows Figures 15A to 15B As shown in the figure, the body weight of the FH-001-treated group remained basically unchanged, while the negative control group showed a significant weight loss since the 4th day after infection, with the maximum decrease exceeding 10%. This result proves that FH-001 has a good preventive and protective effect against RSV virus infection from the perspective of mouse weight changes. Fig. 15B The results show the results of detecting virus titer levels in the nasal and lung tissues of mice on the 6th day after infection. The virus titers in the nose and lungs of the FH-001-treated group were lower than those in the negative control group (approximately 2 times lower). This result proves the good preventive and protective effect of FH-001 against RSV virus from the perspective of virus titer.

[0270] Example 15 Transmission electron microscopy results of different types of nasal spray immune enhancement preparations

[0271] The nasal spray immune enhancement preparations FH-001, Al-001, CaP and biscalcium zinc phosphate were observed by transmission electron microscopy.

[0272] The results are as follows FIG. 16A to FIG. 16D As shown, Fig.16A It is Al-001 form, amorphous and filamentous; Fig. 16B It is the form of FH-001, amorphous and filamentous; Fig. 16C It is in the form of calcium phosphate, cylindrical and rod-shaped, and dispersed relatively evenly; Fig.16D It is the form of calcium zinc bisphosphate, which is irregular polygonal and unevenly dispersed. The difference in the morphology of immune enhancement preparations may lead to differences in their preventive and protective effects.

[0273] Example 16 Nasal spray immune enhancement preparations FH-001, FH-001-Mn, FH-001-Mn + 5% sucrose and FH-001-Mn +5% trehalose sedimentation properties results

[0274] The preparation of FH-001 and FH-001-Mn was as described above. The preparation process of FH-001-Mn + 5% sucrose / trehalose was to mix 4X FH-001-Mn with 10% sucrose / trehalose solution. Figures 17A to 17B As shown, Fig.17AThe four preparations of FH-001, FH-001-Mn, FH-001-Mn+5% sucrose and FH-001-Mn+5% trehalose were white or light yellow suspensions after being shaken. Fig. 17B The appearance of the above four preparations after standing at room temperature for 12 hours. Compared with FH-001, FH-001-Mn has a larger volume of light yellow sediment at the bottom. The volume of the light yellow sediment of the two preparations of FH-001-Mn + 5% sucrose / trehalose is similar.

[0275] Example 17 Nasal spray immune enhancement preparation FH-001 nasal drops against influenza B virus B / Florida / 4 / 2006 Yamagata The relationship between lineage prevention and protection and body immunity

[0276] The mice were randomly divided into three groups, namely, the group receiving nasal drops of the nasal spray immune enhancement preparation FH-001, the group receiving nasal drops of the nasal spray immune enhancement preparation FH-001 after pretreatment with CLL (clodronate liposomes), and the group receiving nasal drops of normal saline. Among them, CLL is an effective macrophage scavenger, and the main function of CLL pretreatment is to clear macrophages. The nasal drops administration volume is 100 μL per mouse, with 1 injection, and the administration is performed on day 0.

[0277] The mice were challenged with isoflurane anesthesia and nasal dripped with 50 μL of 25 LD 50 The B / Florida / 4 / 2006 Yamagata Lineage virus was used to detect the changes in body weight of mice starting from day 7 after administration and continuing for 14 days.

[0278] The results are as follows Figures 18A to 18B As shown in the figure, the weight of mice in the FH-001 administration group was basically not significantly reduced, and the average was still maintained at more than 90%, while the weight of the negative control group (normal saline nasal administration group) decreased significantly after the challenge, with the maximum decrease exceeding 25%, and they basically died on the 8th day after the challenge, which was consistent with the previous results. The mice in the nasal administration group of the newly introduced CLL pretreatment and then nasal spray of the immune enhancement preparation FH-001 in this example lost a significant amount of weight after the lethal challenge of influenza B virus, with a maximum average decrease of about 24%. Fig.18BThe results showed that 14 days after infection, the survival rate of mice in the FH-001 nasal drops group was 100%, all mice in the negative control group died after infection, and no mice survived. The survival rate of mice in the nasal drops group that was first pretreated with CLL and then nasally sprayed with the immune-enhancing agent FH-001 was only 18%. Since the main function of CLL pretreatment is to clear macrophages, according to this result, the preventive and protective effect of FH-001 is related to the existence and function of immune cells, and its preventive and protective effect is mainly achieved by improving the body's immunity.

[0279] Example 18 Immunoenhancing preparation FH-001-Mn combined with SARS-CoV-2 recombinant protein RBD (CHO expression system) Drugs, or immune enhancement preparations Al-001 combined with the new coronavirus recombinant protein RBD (CHO expression system) for the new coronavirus The protective effect of SARS-CoV-2 on mice infected with SARS-CoV-2

[0280] The recombinant protein RBD (receptor-binding domain) of the new coronavirus was produced using the CHO expression system, and then purified by affinity chromatography. The molecular weight and purity (95%) of the protein were determined by the SDS-PAGE system, and the protein concentration in the solution was determined by the bicinchoninic acid (BCA) protein quantification method.

[0281] Balb / C mice were randomly divided into two groups: (1) the group administered with the immune enhancing agent FH-001-Mn combined with the SARS-CoV-2 recombinant protein RBD (CHO expression system); and (2) the group administered with the immune enhancing agent Al-001 combined with the SARS-CoV-2 recombinant protein RBD (CHO expression system). In group (1), FH-001-Mn was mixed with an equal volume of purified SARS-CoV-2 recombinant protein RBD solution, and the mixture was placed at 4°C for at least 30 min before injection into mice. In group (2), Al-001 was mixed with an equal volume of purified SARS-CoV-2 recombinant protein RBD solution, and the mixture was placed at 4°C for at least 30 min before injection into mice. Both groups of mice received three intramuscular injections at weeks 0, 2, and 4, with each injection volume of 150 μL per mouse (containing 10 μg of SARS-CoV-2 recombinant protein RBD), and 75 μL was injected into each hind limb. Blood was collected at weeks 2, 3, 4, 5, and 6 after injection, and the blood was centrifuged at 13,000 rpm for 10 min to separate the serum, which was then stored at 4°C. The serum was measured by enzyme-linked immunosorbent assay (ELISA) using an enzyme-linked immunosorbent assay (ELISA) to obtain the antibody titer in mice. The ELISA method used novel coronavirus antibody detection reagents were purchased from Xiamen Wantai Canghai Biotechnology Co., Ltd., and specifically included the following steps: 1X carbonate buffer (CB, pH = 9.6) was used as a coating solution to coat the novel coronavirus recombinant protein RBD antigen, fixed on a 96-well polystyrene microplate (200 ng per well), and placed at 37°C for 4 hours; 200 μL of blocking solution was added to each well, blocked at 37°C for 2 hours, and then placed at 4°C for standby use. With a starting dilution ratio of 1:100, the mouse serum was serially diluted with enzyme diluent, and the diluent was added to each well, 100 μL per well, incubated at 37°C for 0.5 hours, and the plate was washed 5 times with phosphate buffered saline (PBST) containing Tween-20. Add 100 μL of diluted enzyme-labeled antibody (enzyme-labeled goat anti-mouse antibody (GAM-HRP) diluted with enzyme diluent at a volume ratio of 1:5000) to each well, incubate at 37°C for 0.5h, and wash the plate 5 times with PBST washing solution. Add 100 μL of color development solution (A:B solution mixed in equal volumes) to each well, and react at 37°C in the dark for 10 minutes. Add 50 μL of stop solution to each well to stop the color development process, and use an enzyme reader to measure the absorbance value at 450nm to quantitatively analyze the antibody titer.

[0282] The results are as follows Fig.19 As shown in the figure, it can be seen that both medication groups can produce new coronavirus-specific antibodies and have a preventive effect. Among them, FH-001-Mn combined with the new coronavirus recombinant protein RBD produces new coronavirus-specific antibodies faster, and its immune stimulation effect is better than that of the nasal spray immune enhancement preparation Al-001 combined with the new coronavirus recombinant protein RBD.

[0283] Obviously, the above embodiments are merely examples for the purpose of clear explanation, and are not intended to limit the implementation methods. For those skilled in the art, other different forms of changes or modifications can be made based on the above description. It is not necessary and impossible to list all the implementation methods here. The obvious changes or modifications derived therefrom are still within the scope of protection of the invention.

Claims

1. A method for preparing a pharmaceutical mixture, wherein: The medicinal mixture is used to prevent respiratory tract transmission virus infection or diseases or conditions caused by it, and the respiratory tract transmission virus is influenza B virus; The method comprises the following steps: (1) providing a soluble salt solution; the cations in the soluble salt solution are zinc ions; or the cations in the soluble salt solution are zinc ions and calcium ions, and the molar ratio of zinc ions to calcium ions is 1:(0.4-3); (2) providing an alkaline solution comprising phosphate ions, bisphosphonate ions and hydroxide ions; wherein the molar ratio of the phosphate ions, the bisphosphonate ions and the hydroxide ions in the alkaline solution is (4-7):1:(3-11); (3) Mixing the soluble salt solution and the alkaline solution in a volume ratio of 1:(0.5-2) to obtain a mixture.

2. The method according to claim 1, wherein: In step (3), the alkaline solution is added to the soluble salt solution for mixing.

3. The method according to claim 1, further comprising step (4): sterilizing the mixture to obtain a sterilized product.

4. The method according to claim 1 or 3, further comprising step (5): adjusting the pH value of the mixture or the sterilized product to 6-7.

5. The method according to claim 1, characterized in that One or more of the following: (A) In step (1), the molar concentration of zinc ions in the soluble salt solution is 20 to 83 mM; (B) In step (2), the molar concentration of phosphate ions in the alkaline solution is 20 to 65 mM; (C) The bisphosphonate ion is selected from the group consisting of a zoledronic acid ion and a risedronate ion.

6. A method for preparing a pharmaceutical mixture, wherein: The medicinal mixture is used to prevent respiratory tract-transmitted virus infection or diseases or conditions caused by it, and the respiratory tract-transmitted virus is a new coronavirus; The method comprises the following steps: 1) providing a soluble salt solution containing zinc ions, aluminum ions and manganese ions; wherein the molar ratio of zinc ions, aluminum ions and manganese ions in the soluble salt solution is 1:(1-2):1; 2) providing an alkaline solution comprising phosphate ions and hydroxide ions; wherein the molar concentration of the phosphate ions in the alkaline solution is 12 to 24 mM, and the molar concentration of the hydroxide ions is 130 to 160 mM; 3) Mixing the soluble salt solution and the alkaline solution in a volume ratio of 1:(0.5-1.5) to obtain a mixture.

7. The method according to claim 6, characterized in that One or more of the following: A. The molar concentration of zinc ions in the soluble salt solution is 20-64 mM; B. In step 3), the alkaline solution is added to the soluble salt solution for mixing; C. The method further comprises step 4): sterilizing the mixture to obtain a sterilized product.

8. The method according to claim 7, wherein: In step 4), the pH value of the sterilized product is adjusted to 6-7.

9. A pharmaceutical mixture prepared by the method according to any one of claims 1 to 5; the pharmaceutical mixture is used to prevent respiratory tract-transmitted virus infection or diseases or conditions caused by it, wherein the respiratory tract-transmitted virus is influenza B virus.

10. The drug mixture according to claim 9, comprising a plurality of selected from the group consisting of calcium chloride, calcium phosphate, calcium zoledronic acid, calcium risedronate, zinc chloride, zinc phosphate, zinc hydroxide, zinc zoledronic acid and zinc risedronate.

11. A pharmaceutical mixture prepared by the method according to any one of claims 6 to 8; the pharmaceutical mixture is used to prevent respiratory tract-transmitted virus infection or diseases or symptoms caused by it, wherein the respiratory tract-transmitted virus is a new coronavirus.

12. The pharmaceutical mixture according to claim 11, comprising a plurality of compounds selected from the group consisting of ZnCl2, AlCl3, MnCl2, Zn(OH)2, Al(OH)3, Mn(OH)2, AlPO4 and aluminum hydroxyphosphate.

13. A pharmaceutical preparation comprising the pharmaceutical mixture according to any one of claims 9 to 12 and optional pharmaceutical excipients. The pharmaceutical preparation according to claim 13, which is a nasal spray preparation or a nasal drop preparation.

15. Use of a pharmaceutical mixture in the preparation of a medicament for preventing respiratory tract-transmitted viral infections, or in the preparation of a medicament for preventing diseases or conditions caused by respiratory tract-transmitted viral infections; wherein: The respiratory virus is selected from influenza A virus, influenza B virus, respiratory syncytial virus and new coronavirus; Wherein, the pharmaceutical mixture is the pharmaceutical mixture according to any one of claims 9 to 12, or the pharmaceutical mixture is prepared by a method comprising the following steps: i) providing a soluble salt solution comprising at least one selected from zinc ions, aluminum ions and calcium ions; ii) providing an alkaline solution comprising phosphate ions and hydroxide ions; iii) mixing the soluble salt solution with the alkaline solution to obtain a mixture, wherein the volume ratio of the soluble salt solution to the alkaline solution is 1:(0.5-2); And, including any of the following: a. The soluble salt solution contains zinc ions and aluminum ions; the molar ratio of zinc ions to aluminum ions in the soluble salt solution is 1:(1-4); the molar ratio of phosphate ions to hydroxide ions in the alkaline solution is 1:(8-14); b. The soluble salt solution contains aluminum ions but does not contain calcium ions and zinc ions; the molar ratio of phosphate ions to hydroxide ions in the alkaline solution is 1:(6-11); c. The soluble salt solution contains calcium ions but does not contain zinc ions and aluminum ions; the molar ratio of phosphate ions to hydroxide ions in the alkaline solution is (1-7):

1.

16. The use according to claim 15, wherein In step iii), mixing is carried out by adding the alkaline solution to the soluble salt solution.

17. The use according to claim 15, in item a, characterized in that One or more of the following: The molar concentration of zinc ions in the soluble salt solution is 20 to 64 mM; The molar concentration of phosphate ions in the alkaline solution is 4 to 30 mM.

18. The use according to claim 15, in item b, characterized in that One or more of the following: The molar concentration of aluminum ions in the soluble salt solution is 40-80 mM; The molar concentration of phosphate ions in the alkaline solution is 3 to 20 mM.

19. The use according to claim 15, in item c, characterized in that One or more of the following: The molar concentration of calcium ions in soluble salt solutions is 40 to 82 mM; The molar concentration of phosphate ions in alkaline solution is 40-82 mM.

20. The use according to claim 15, characterized in that One or more of the following: (a) a disease or condition caused by the respiratory virus infection selected from rhinitis, pharyngitis, laryngitis, bronchitis, pneumonia, fever, headache, pharyngitis, fatigue, muscle aches, nausea and vomiting, dry cough, dyspnea, nasal congestion, runny nose and diarrhea; (b) the drug is in the form of a nasal spray or nasal drops; (c) The method further comprises step iv): sterilizing the mixture to obtain a sterilized product.

21. The use according to claim 15, wherein The new coronavirus is SARS-CoV-2.

22. The use according to claim 20, wherein In step iv), the pH value of the sterilized product is adjusted to 6-7.

23. A pharmaceutical preparation comprising a medicinal mixture, a recombinant protein RBD of a novel coronavirus, and optional pharmaceutical excipients, wherein the volume ratio of the medicinal mixture to the liquid containing the recombinant protein RBD of the novel coronavirus is 1:1, and the content of the recombinant protein RBD of the novel coronavirus in the liquid is 1 / 15 μg / μL; wherein The pharmaceutical mixture is the pharmaceutical mixture according to claim 11 or 12, or the pharmaceutical mixture is prepared by a method comprising the following steps: 1) providing a soluble salt solution containing aluminum ions; wherein, The molar concentration of aluminum ions in the soluble salt solution is 50-65 mM; II) providing an alkaline solution comprising phosphate ions and hydroxide ions; wherein the molar ratio of the phosphate ions to the hydroxide ions in the alkaline solution is 1:(6.7-9.4); III) mixing the soluble salt solution and the alkaline solution in a volume ratio of 1:(0.5-1.5) to obtain a mixture.

24. The pharmaceutical preparation according to claim 23, wherein In step II), the molar concentration of phosphate ions in the alkaline solution is 3-20 mM.

25. Use of a medicinal mixture in combination with a novel coronavirus recombinant protein RBD in the preparation of a drug for preventing novel coronavirus infection; in, The pharmaceutical mixture is the pharmaceutical mixture as claimed in claim 23 or 24; The volume ratio of the medicinal mixture to the liquid containing the new coronavirus recombinant protein RBD is 1:1, and the content of the new coronavirus recombinant protein RBD in the liquid is 1 / 15 μg / μL.

26. The use according to claim 25, wherein The new coronavirus is SARS-CoV-2.

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