A solution-type nasal spray for treating rhinitis and a method of preparation

Abstract

The application discloses a solution type nasal spray containing 3-[(2-cyclopentyl-2-hydroxy-2-phenyl)ethoxy]-1-(3-phenoxypropyl)-1-azabicyclo[2,2,2]octane or a derivative thereof and a preparation method of the nasal spray. The nasal spray contains a main drug, a solvent, a cosolvent, a bacteriostatic agent, a pH regulator and an osmotic pressure regulator. The nasal spray has good stability, the pharmacodynamic test result shows that the nasal spray has fast effect and small irritation, and can be used for treating various rhinitis and cold accompanied rhinitis.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of pharmaceutical preparation technology, and particularly relates to a solution type nasal spray for treating various rhinitis and cold accompanied rhinitis and a preparation method thereof. BACKGROUND

[0002] Rhinitis includes acute rhinitis such as cold accompanied rhinitis and chronic rhinitis. Chronic rhinitis is classified as allergic rhinitis (AR) and non-allergic rhinitis (NAR) according to allergens, and is classified as seasonal rhinitis and perennial rhinitis according to the onset time. Rhinitis is a common respiratory disease, among which AR is one of the most common diseases in adults, and the incidence shows an obvious increasing trend. The typical symptoms of rhinitis are sneezing, rhinorrhea, nasal itching and nasal congestion.

[0003] At present, the drugs for treating rhinitis mainly include glucocorticoids, antihistamines and anticholinergic drugs, and the preparation is mainly nasal spray. Glucocorticoids and antihistamines have certain effects on rhinorrhea, nasal congestion, sneezing and nasal itching, among which glucocorticoids have a slow effect, and generally need more than 2 weeks to obtain good efficacy, and the efficacy on AR with negative eosinophils is not significant. In addition, oral and intranasal administration of antihistamines has uncertain efficacy on NAR, and both hormones and antihistamines have certain side effects.

[0004] The anticholinergic drug ipratropium bromide (IPR) nasal spray can treat the symptoms of rhinitis such as cold accompanied rhinitis, AR and NAR, and can be used for short-term or long-term treatment. Its advantages are fast effect and less side effects, and it can be used as an urgently needed drug for symptomatic treatment, but the action time is short, and the patient needs to be given 3-4 times or even more per day, and the patient compliance is poor. The long-acting anticholinergic drug (LAMA) that has been marketed is not suitable for the treatment of rhinitis because it has no selectivity for M receptor subtypes. The compound of formula I has strong selectivity for M receptor subtypes, is an ideal LAMA for treating rhinitis, has the characteristics of strong and lasting effect on M3 receptors and very weak and short duration of effect on M2, and is intended to be used for treating symptoms such as rhinorrhea and nasal itching caused by various rhinitis. Through years of research, the pharmacological effects of the compound of formula I have been fully proved. The nasal spray of the present application achieves the purposes of good stability, improved drug concentration and elimination of irritation through prescription and process innovation compared with the prior art; the nasal spray of the present application has the characteristics of faster effect compared with the nasal spray of the prior art, and produces unexpected effects. SUMMARY

[0005] The present application provides a nasal spray which can be used for treating acute and chronic rhinitis, and is in the form of a clear and uniform solution. The nasal spray comprises a compound of formula I, water, a cosolvent, a bacteriostatic agent, a pH adjusting agent and a tonicity adjusting agent NaCl.

[0006] The active ingredient in the present application is compound of formula I

[0007]

[0008] M - Suitable organic or inorganic pharmaceutically acceptable anions can be selected from, but not limited to, bromide, fluoride, chloride, iodide, nitrate, sulfate, phosphate, formate, acetate, trifluoroacetate, propionate, butyrate, lactate, citrate, tartrate, malate, maleate, succinate, benzoate, p-chlorobenzoate, diphenylacetate, triphenylacetate, o-hydroxybenzoate, p-hydroxybenzoate, 1-hydroxynaphthalene-2-carboxylate, 3-hydroxynaphthalene-2-carboxylate, methanesulfonate or p-toluenesulfonate. Compound of formula I has two chiral centers, thus there are four optical isomers, i.e. compounds of (2R, 3R), (2S, 3R), (2R, 3S) and (2S, 3S) configurations. The compound of formula I described in the present application also contains one or more of the above isomers, or their mixtures and racemates, such as (2S / 3S, 2R / 3R) racemate, which are all within the scope of the present application.

[0009] A representative compound of formula I is compound 1, having the molecular formula C 29 H 40 BrNO3, with a molecular weight of 530.54, and having the structure as follows:

[0010]

[0011] M - As a pharmaceutically acceptable anion, the preparation of the compound is known in the prior art, see patent document WO2015007073.

[0012] In the nasal spray of the present application, the co-solvent includes one or more than one mixture of ethanol, glycerol, glycols, wherein the glycols include but are not limited to ethylene glycol, propylene glycol, polyethylene glycol 200, polyethylene glycol 300, polyethylene glycol 400, polyethylene glycol 600, polyethylene glycol 800, etc.

[0013] In the nasal spray of the present application, the bacteriostatic agent is benzalkonium chloride or benzalkonium bromide; the structures of benzalkonium chloride and benzalkonium bromide are shown in formula II, X is Cl for benzalkonium chloride, X is Br for benzalkonium bromide, and n is between 1 and 18. There are pure single molecules and mixtures of different molecules in the market, and the literature shows that there is no significant difference in bacteriostatic effect.

[0014]

[0015] The content of the compound of formula I in the nasal spray of the present application is 0.01-1% by weight, preferably 0.02-0.5% by weight; the content of the cosolvent is 0-10% by weight, preferably 0-5% by weight; the content of the bacteriostatic agent in the nasal spray is 0.001-0.05% by weight, preferably 0.005-0.02% by weight. The content of the osmotic pressure regulator NaCl in the nasal spray is 0.45-2.25% by weight, preferably 0.45-1.35% by weight. The pH of the nasal spray is 4.5-8, preferably 5-7.

[0016] The nasal spray of the present application can further improve the drug content, increase the stability, reduce the irritation, and have a fast onset of action compared with the prior art. The animal overall anti-rhinitis test uses an ovalbumin (OVA)-induced guinea pig allergic rhinitis model to observe the effect of the compound of formula I on the symptoms of the allergic rhinitis model and the onset time; the guinea pig nose is continuously administered for 30 days to observe the irritation of the compound of formula I nasal spray pH, NaCl content, and cosolvent to the nasal mucosa. The nasal spray of the present application can be used for treating various rhinitis, including acute rhinitis such as cold-associated rhinitis and chronic rhinitis. Chronic rhinitis is classified as allergic rhinitis (AR) and non-allergic rhinitis (NAR) according to the allergen, and is classified as seasonal rhinitis and perennial rhinitis according to the onset time, and the typical symptoms of cold-associated rhinitis and AR are symptoms such as runny nose and nasal itching.

[0017] Another object of the present application is to provide a preparation method of a nasal spray, which comprises the steps of dissolving the compound of formula I or its optical isomer and racemate, cosolvent, bacteriostatic agent, NaCl, etc. with water, adjusting to a suitable pH with an acid, salt, or base, filling the finished product into a glass or plastic bottle, and adding a spray head and a protective cap. The filling method belongs to the conventional technology in the art and is described in textbooks in the art, which are incorporated by reference into the present application. DETAILED DESCRIPTION

[0018] The present application is further described below through specific embodiments. It should be noted that the following specific embodiments are only used to illustrate the present application, and those skilled in the art can make corresponding changes to the present application according to the prior art and common knowledge in the technical field, and these technical solutions all fall within the scope of the present application.

[0019] The compound synthesized according to the patent document with publication number WO2015007073 is as follows:

[0020] Compound 1: (2R, 3R)-3-[(2-cyclopentyl-2-hydroxy-2-phenyl)ethoxy]-1-(3-phenoxypropyl)-1-azabicyclo[2, 2, 2]octane bromide;

[0021] Compound 2: (2R,3R)-3-[(2-cyclopentyl-2-hydroxy-2-phenyl)ethoxy]-1-(3-phenoxypropyl)-1-azabicyclo[2,2,2]octanium fumarate (1:1) salt;

[0022] Compound 3: (2R,3R)-3-[(2-cyclopentyl-2-hydroxy-2-phenyl)ethoxy]-1-(3-phenoxypropyl)-1-azabicyclo[2,2,2]octanium methanesulfonate;

[0023] Compound 4: (2R,3S),(2S,3R)-3-[(2-cyclopentyl-2-hydroxy-2-phenyl)ethoxy]-1-(3-phenoxypropyl)-1-azabicyclo[2,2,2]octanium bromide;

[0024] Compound 5: (2R,3S),(2S,3R)-3-[(2-cyclopentyl-2-hydroxy-2-phenyl)ethoxy]-1-(3-phenoxypropyl)-1-azabicyclo[2,2,2]octanium chloride;

[0025] Compound 6: 3-[(2-cyclopentyl-2-hydroxy-2-phenyl)ethoxy]-1-(3-phenoxypropyl)-1-azabicyclo[2,2,2]octanium bromide;

[0026] Compound 7: (2R,3R),(2S,3S)-3-[(2-cyclopentyl-2-hydroxy-2-phenyl)ethoxy]-1-(3-phenoxypropyl)-azabicyclo[2,2,2]octanium dihydrogen phosphate;

[0027] Compound 8: (2S,3R)-3-[(2-cyclopentyl-2-hydroxy-2-phenyl)ethoxy]-1-(3-phenoxypropyl)-1-azabicyclo[2,2,2]octanium chloride maleate (1:1) salt;

[0028] Compound 9: (2S,3R)-3-[(2-cyclopentyl-2-hydroxy-2-phenyl)ethoxy]-1-(3-phenoxypropyl)-1-azabicyclo[2,2,2]octanium tartrate (1:1) salt;

[0029] Compound 10: (2R,3S)-3-[(2-cyclopentyl-2-hydroxy-2-phenyl)ethoxy]-1-(3-phenoxypropyl)-1-azabicyclo[2,2,2]octanium hydrogen sulfate.

[0030] Example 1

[0031] Specification: 140 μl per spray, containing 20 μg of compound 1

[0032] Prescription component Amount Action Implementation standard Compound 1 (as anhydride) 1.43 mg Main drug Enterprise standard Benzalkonium chloride 1.00 mg Antiseptic bacteriostatic agent Chinese Pharmacopoeia 2020 edition Ⅱ Sodium chloride 90.00 mg Osmotic pressure regulator Chinese Pharmacopoeia 2020 edition Ⅳ Disodium hydrogen phosphate 0.60 mg pH regulator Chinese Pharmacopoeia 2020 edition Ⅳ Hydrochloric acid Sufficient amount to adjust pH to 5.0 pH regulator Chinese Pharmacopoeia 2020 edition Ⅳ Purified water To 10 ml Solvent Chinese Pharmacopoeia 2020 edition Ⅱ

[0033] Example 2

[0034] Specification: 140 μl per spray, containing Compound 1 80 μg

[0035] Prescription component Amount Action Implementation standard Compound 1 (as anhydride) 5.72 mg Main drug Enterprise standard Benzalkonium chloride 1.00 mg Antiseptic bacteriostatic agent Chinese Pharmacopoeia 2020 edition Ⅱ Sodium chloride 90.00 mg Osmotic pressure regulator Chinese Pharmacopoeia 2020 edition Ⅳ Disodium hydrogen phosphate 0.60 mg pH regulator Chinese Pharmacopoeia 2020 edition Ⅳ Hydrochloric acid Sufficient amount to adjust pH to 6.0 pH regulator Chinese Pharmacopoeia 2020 edition Ⅳ Purified water To 10 ml Solvent Chinese Pharmacopoeia 2020 edition Ⅱ

[0036] Example 3

[0037] Specification: 140 μl per spray, containing Compound 1 240 μg

[0038] Prescription component Amount Action Implementation standard Compound 1 (as anhydride) 17.16 mg Main drug Enterprise standard Benzalkonium chloride 1.00 mg Antiseptic bacteriostatic agent Chinese Pharmacopoeia 2020 edition Ⅱ Sodium chloride 90.00 mg Osmotic pressure regulator Chinese Pharmacopoeia 2020 edition Ⅳ Disodium hydrogen phosphate 0.60 mg pH regulator Chinese Pharmacopoeia 2020 edition Ⅳ Hydrochloric acid Sufficient amount to adjust pH to 6.2 pH regulator Chinese Pharmacopoeia 2020 edition Ⅳ Purified water To 10 ml Solvent Chinese Pharmacopoeia 2020 edition Ⅱ

[0039] Example 4

[0040] Specification: 90 μl per spray, containing Compound 1 80 μg

[0041] Prescription component Amount Action Implementation standard Compound 1 (as anhydride) 8.89 mg Main drug Enterprise standard Benzalkonium chloride 1.00 mg Antiseptic bacteriostatic agent Chinese Pharmacopoeia 2020 edition Ⅱ Sodium chloride 90.00 mg Osmotic pressure regulator Chinese Pharmacopoeia 2020 edition Ⅳ Disodium hydrogen phosphate 0.60 mg pH regulator Chinese Pharmacopoeia 2020 edition Ⅳ Hydrochloric acid Sufficient amount to adjust pH to 6.5 pH regulator Chinese Pharmacopoeia 2020 edition Ⅳ Purified water To 10 ml Solvent Chinese Pharmacopoeia 2020 edition Ⅱ

[0042] Example 5

[0043] Specification: 140 μl per spray, containing Compound 2 80 μg

[0044] Prescription component Amount Action Implementation standard Compound 2 (as anhydride) 5.72 mg Main drug Enterprise standard Benzalkonium chloride 2.00 mg Antiseptic bacteriostatic agent Chinese Pharmacopoeia 2020 edition Ⅱ Sodium chloride 45.00 mg Osmotic pressure regulator Chinese Pharmacopoeia 2020 edition Ⅳ Sodium hydrogen fumarate 10.00 mg pH regulator Chinese Pharmacopoeia 2020 edition Ⅳ Hydrochloric acid Sufficient amount to adjust pH to 7.0 pH regulator Chinese Pharmacopoeia 2020 edition Ⅳ Purified water To 10 ml Solvent Chinese Pharmacopoeia 2020 edition Ⅱ

[0045] Example 6

[0046] Specification: 140 μl per spray, containing Compound 3 240 μg

[0047] Prescription component Amount Action Implementation standard Compound 3 (as anhydride) 17.16 mg Main drug Enterprise standard Benzalkonium chloride 5.00 mg Antiseptic bacteriostatic agent Chinese Pharmacopoeia 2020 edition Ⅱ Sodium chloride 45.00 mg Osmotic pressure regulator Chinese Pharmacopoeia 2020 edition Ⅳ Disodium hydrogen phosphate 6.00 mg pH regulator Chinese Pharmacopoeia 2020 edition Ⅳ Hydrochloric acid Sufficient amount to adjust pH to 4.0 pH regulator Chinese Pharmacopoeia 2020 edition Ⅳ Purified water To 10 ml Solvent Chinese Pharmacopoeia 2020 edition Ⅱ

[0048] Example 7

[0049] Specification: 140 μl per spray, containing Compound 4 240 μg

[0050] Prescription component Amount Action Implementation standard Compound 4 (as anhydrous substance) 17.16 mg Main drug Enterprise standard Benzalkonium chloride 3.00 mg Antiseptic bacteriostatic agent Chinese Pharmacopoeia 2020 edition part two Sodium chloride 90 mg Osmotic pressure regulator Chinese Pharmacopoeia 2020 edition part two Ethylene glycol 500 mg Solubilizer Chinese Pharmacopoeia 2020 edition part four Disodium hydrogen phosphate 6.00 mg pH regulator Chinese Pharmacopoeia 2020 edition part four Hydrochloric acid Sufficient amount to adjust pH to 6.1 pH regulator Chinese Pharmacopoeia 2020 edition part four Purified water To 10 ml Solvent Chinese Pharmacopoeia 2020 edition part two

[0051] Example 8

[0052] Specification: 140 μl per spray, containing Compound 5 700 μg

[0053] Prescription components Dosage Action Execution standard Compound 5 (as anhydrous substance) 50.00 mg Main drug Enterprise standard Benzalkonium chloride 1.00 mg Antiseptic bacteriostatic agent Chinese Pharmacopoeia 2020 edition part two Sodium chloride 45 mg Osmotic pressure regulator Chinese Pharmacopoeia 2020 edition part two Propylene glycol 1000 mg Solubilizer Chinese Pharmacopoeia 2020 edition part four Sodium dihydrogen phosphate 5.00 mg pH regulator Chinese Pharmacopoeia 2020 edition part four Sodium hydroxide Sufficient amount to adjust pH to 8.0 pH regulator Chinese Pharmacopoeia 2020 edition part four Purified water To 10 ml Solvent Chinese Pharmacopoeia 2020 edition part two

[0054] Example 9

[0055] Specification: 140 μl per spray, containing Compound 6 240 μg

[0056] Prescription components Dosage Action Execution standard Compound 6 (as anhydrous substance) 17.16 mg Main drug Enterprise standard Benzalkonium chloride 1.00 mg Antiseptic bacteriostatic agent Chinese Pharmacopoeia 2020 edition part two Sodium chloride 90.00 mg Osmotic pressure regulator Chinese Pharmacopoeia 2020 edition part two Ethylene glycol 800 500 mg Solubilizer Chinese Pharmacopoeia 2020 edition part four Disodium hydrogen phosphate 5.00 mg pH regulator Chinese Pharmacopoeia 2020 edition part four Hydrochloric acid Sufficient amount to adjust pH to 6.0 pH regulator Chinese Pharmacopoeia 2020 edition part four Purified water To 10 ml Solvent Chinese Pharmacopoeia 2020 edition part two

[0057] Example 10

[0058] Specification: 140 μl per spray, containing Compound 6 420 μg

[0059] Prescription components Dosage Action Execution standard Compound 6 (as anhydrous substance) 30.00 mg Main drug Enterprise standard Benzalkonium chloride 0.10 mg Antiseptic bacteriostatic agent Chinese Pharmacopoeia 2020 edition part two Sodium chloride 90.00 mg Osmotic pressure regulator Chinese Pharmacopoeia 2020 edition part two Ethylene glycol: ethanol (1:1) 1000 mg Solubilizer Chinese Pharmacopoeia 2020 edition part four Disodium hydrogen phosphate 1.00 mg pH regulator Chinese Pharmacopoeia 2020 edition part four Hydrochloric acid Sufficient amount to adjust pH to 4.5 pH regulator Chinese Pharmacopoeia 2020 edition part four Purified water To 10 ml Solvent Chinese Pharmacopoeia 2020 edition part two

[0060] Example 11

[0061] Specification: 140 μl per spray, containing compound 7 220 μg

[0062] Prescription components Dosage Action Execution standard Compound 7 (as anhydrous substance) 15.00 mg Main drug Enterprise standard Benzalkonium chloride 1.00 mg Antiseptic bacteriostatic agent Chinese Pharmacopoeia 2020 edition part two Sodium chloride 90.00 mg Osmotic pressure regulator Chinese Pharmacopoeia 2020 edition part two Ethylene glycol 300 mg Solubilizer Chinese Pharmacopoeia 2020 edition part four Disodium hydrogen phosphate 0.60 mg pH regulator Chinese Pharmacopoeia 2020 edition part four Hydrochloric acid Sufficient amount to adjust pH to 6.0 pH regulator Chinese Pharmacopoeia 2020 edition part four Purified water To 10 ml Solvent Chinese Pharmacopoeia 2020 edition part two

[0063] Example 12

[0064] Specification: 90 μl per spray, containing compound 8 135 μg

[0065] Prescription components Dosage Action Execution standard Compound 8 (as anhydrous substance) 15.00 mg Main drug Enterprise standard Benzalkonium chloride 1.00 mg Antiseptic bacteriostatic agent Chinese Pharmacopoeia 2020 edition part two Sodium chloride 90.00 mg Osmotic pressure regulator Chinese Pharmacopoeia 2020 edition part two Osmotic pressure regulator Chinese Pharmacopoeia 2020 edition volume 4 Disodium hydrogen phosphate 0.60 mg pH regulator Chinese Pharmacopoeia 2020 edition volume 4 Hydrochloric acid Sufficient amount to adjust pH to 5.5 pH regulator Chinese Pharmacopoeia 2020 edition volume 4 Purified water To 10 ml Solvent Chinese Pharmacopoeia 2020 edition volume 2

[0066] Example 13

[0067] Specification: 90 μl per spray, containing compound 9 900 μg

[0068] Prescription components Dosage Effects Execution standards Compound 9 (as anhydride) 100.00 mg Main drug Enterprise standard Benzalkonium bromide 1.00 mg Antiseptic bacteriostatic agent Chinese Pharmacopoeia 2020 edition volume 2 Sodium chloride 90.00 mg Osmotic pressure regulator Chinese Pharmacopoeia 2020 edition volume 4 Glycerin 500 mg Solubilizer Chinese Pharmacopoeia 2020 edition volume 4 Sodium hydrogen tartrate 0.60 mg pH regulator Chinese Pharmacopoeia 2020 edition volume 4 Hydrochloric acid Sufficient amount to adjust pH to 7.5 pH regulator Chinese Pharmacopoeia 2020 edition volume 4 Purified water To 10 ml Solvent Chinese Pharmacopoeia 2020 edition volume 2

[0069] Example 14

[0070] Specification: 90 μl per spray, containing compound 10 450 μg

[0071] Prescription components Dosage Effects Execution standards Compound 10 (as anhydride) 50.00 mg Main drug Enterprise standard Benzalkonium bromide 1.00 mg Antiseptic bacteriostatic agent Chinese Pharmacopoeia 2020 edition volume 2 Sodium chloride 90.00 mg Osmotic pressure regulator Chinese Pharmacopoeia 2020 edition volume 4 Ethanol 500 mg Solubilizer Chinese Pharmacopoeia 2020 edition volume 4 Disodium hydrogen phosphate 0.60 mg pH regulator Chinese Pharmacopoeia 2020 edition volume 4 Sulfuric acid Sufficient amount to adjust pH to 6.2 pH regulator Chinese Pharmacopoeia 2020 edition volume 4 Purified water To 10 ml Solvent Chinese Pharmacopoeia 2020 edition volume 2

[0072] Determination of drug content and related substances in the sample of Example 1

[0073] (1) Method

[0074] The drug solution of Examples 1-14 was a clear liquid, which was filtered through a 0.22 μm or 0.45 μm micropore filter. The drug content of the filtrate was the same as that of the solution before filtration, as determined by HPLC.

[0075] The chromatographic conditions of the HPLC content determination method were as follows:

[0076]

[0077] Preparation of Assay Control Solution: The assay control solution was prepared by accurately weighing a suitable amount of the corresponding compound of Formula I as the control, dissolving it in the mobile phase. The sample was diluted with the mobile phase to obtain a solution containing 2 μg per 1 ml. The retention time of the various optical isomers of the compound of Formula I was as follows: the retention time of the RR or SS isomers was about 15 min (Rf = 1), and the two isomers coincided; the retention time of the (2R, 3S) and (3S, 2R) isomers was about 1.07 (Rf = 1.07), and the diastereoisomers were well separated from the main drug as impurities. The enantiomers as impurities had the same retention time as the main drug, and must be separated using a chiral column. The results showed that the content of the enantiomers was less than 0.02% of the main drug (not published); the retention time of the acid radical, bromine and solvent peaks was about 0.13-0.2 min; the retention time of benzalkonium chloride (or bromide) (Formula II) was about 3 (n = 2), and the retention time of the degradation product thereof was about 1.27; the retention time of the leachate was about 2.2; the retention time of the maximum impurity introduced during the synthesis of the raw material was about 0.53, and the content was less than 0.01%; and the above components were well separated. The impurities introduced during the preparation of the drug and the optical isomers did not change significantly during long-term storage and long-term sample retention, and were controlled as special known impurities. The impurities produced during the preparation and storage of the preparation included the degradation products of the bacteriostatic agent and the leachate of the inner package, which were controlled as unknown impurities.

[0078] (2) Results

[0079] ① The impurities of different compounds of Formula I and preparations at 0 days are shown in Table 1:

[0080] Table 1. Impurity content (%) of different compounds of Formula I and preparations at 0 days of production

[0081]

[0082] Table 1 shows that the total impurity control of different compounds of Formula I is very good. "*"": the main drug component; "-": the impurity is part of the main drug or the impurity contains the main drug

[0083] ② Stability study of compounds of Formula I under different pH conditions

[0084] The total impurities of Examples 1, 2, 5, 6, 9 and 10 were determined by the method of Example 1 under different pH conditions at 60°C. The results are shown in Table 2:

[0085] Table 2. Total impurity results of compounds of Formula I in aqueous solutions at different pH stored at 60°C for 24 h

[0086]

[0087] * The most stable pH of the solution of compound 1 of Formula I

[0088] Table 2 shows the stability at pH 4-8, the total impurities increase slightly at 60°C for 24 hours at pH 5-7, especially at pH 6.2, the increase is the least; when the pH is greater than 7 or less than 5, the total impurities increase obviously at 60°C for 10 days.

[0089] ③Stability of the nasal spray of the examples under high temperature, strong light irradiation and accelerated conditions

[0090] Under high temperature (50°C) and accelerated conditions (40°C / 75% RH), the samples were determined for the related substances by the method of Experimental Example 1, and the results are shown in Tables 3-4.

[0091] Table 3, results of the high temperature test of the related substances of the nasal spray of the examples (stored at 50°C)

[0092]

[0093] Table 4, results of the accelerated test of the related substances of the nasal spray of the examples (stored at 40°C / 75% RH)

[0094]

[0095] Conclusion: The total related substances of the nasal spray of the examples increase less or not obviously under normal conditions when the samples are placed at 50°C for 5 or 10 days or at 40°C / 75% RH for 6 months. However, the total impurities increase obviously when the pH is less than 5 or greater than 7.

[0096] ④Effect of the concentration of the compound on the stability of the nasal spray during the storage and the effect of the cosolvent

[0097] It is found in the experiment that the solubility of the compound of Formula I in pure water is about 0.25%, but it can decrease to less than 0.2% during the storage. It is analyzed that the drug solution close to the saturated concentration is adsorbed by the inner package (glass bottle, plastic bottle or spray head). After selection, it is found that ethanol, glycerol, isopropyl alcohol, ethylene glycol, polyethylene glycol 200, polyethylene glycol 400, polyethylene glycol 600, polyethylene glycol 800 or the mixture of these alcohols can effectively increase the solubility. When the concentration of the cosolvent is increased to 3%, the drug content can be increased to 0.5%; when the concentration of the cosolvent is increased to 5%, the drug content can be increased to more than 1%. The solution with the cosolvent is very stable, which can meet the requirements of various uses of the nasal spray.

[0098] Experimental Example 2, effect of pH on the therapeutic effect and onset time of the nasal scratching symptoms of the guinea pig allergic rhinitis model induced by ovalbumin (OVA)

[0099] (1) Method

[0100] Sensitization and challenge: Sensitization on Day 1, 3, 5, 7, 14, 21 by intraperitoneal injection of OVA + aluminum hydroxide gel; Challenge on Day 28-34 by intranasal administration of OVA solution, once a day, for a total of 7 days.

[0101] Grouping and administration: Guinea pigs were randomly grouped, 8 in each group, half male and half female, into Control, Model, and administration groups with pH values of 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, and 8, respectively, and other components being the same as in Example 2. The guinea pigs were administered 30 min after challenge on Day 34, with a total volume of 20 μl / 300 g of body weight, and the same amount was administered to both nostrils.

[0102] Index detection: The effect of the test drug on nasal mucosal inflammation was mainly observed. On Day 34, the guinea pigs were administered 30 min after challenge, and the number of times the guinea pigs scratched their noses was observed at 0-10 min, 10-20 min, 20-30 min, 30-40 min, 40-50 min, and 50-60 min after administration, and statistical analysis was performed.

[0103] (2) Results

[0104] ① The results of inhibiting the symptoms of allergic rhinitis in guinea pigs induced by OVA are shown in Table 5:

[0105] Table 5, Effect on the scratching of the nose of guinea pigs with allergic rhinitis induced by OVA (x ± SEM)

[0106]

[0107] Statistics: one way ANOVA, compared with the Control group: ###p < 0.001; compared with the Model group: *p < 0.05

[0108] Table 5 shows that the different pH values of the nasal spray of the present application have a significant inhibitory effect on the itching of the nose of guinea pigs with rhinitis induced by OVA, and the pH value has a significant effect on the onset time. When the pH value is less than 5 (4 and 4.5) or greater than 7 (7.5 and 8), the significant effect is achieved 20 min after administration (*p < 0.05); when the pH value is between 5 and 7, the significant effect is achieved within 20 min (pH 5 and 5.5) or even within 10 min (pH 6, 6.5, and 7) after administration (*p < 0.05).

[0109] Experimental Example 3, Stimulatory effect of the pH, NaCl content, and cosolvent of the nasal spray on the nasal mucosa of guinea pigs

[0110] (1) Methods

[0111] The guinea pigs were divided into groups by random method, half male and half female, 8 in each group, divided into control group and test drug group, the control group was given 0.9% NaCl solution by nasal drops; the test drug group was tested for different pH, different NaCl concentration, and different types of cosolvent, the drug volume of the animals was 20 μl / 300 g body weight, the same amount was given to the left and right nostrils, twice a day, for 30 days.

[0112] The H&E staining of nasal mucosa inflammation pathological grading score standard is shown in Table 6:

[0113] Table 6, H&E staining of nasal mucosa inflammation pathological grading score standard

[0114]

[0115] (2) Results

[0116] ①The influence of pH: the pH of the test drug nasal spray was selected as 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5 and 8, and other components were the same as in Example 2, the irritation of different pH to the nasal mucosa of guinea pigs is shown in Table 7:

[0117] Table 7, the irritation of different pH to the nasal mucosa of guinea pigs (x±SEM)

[0118] Group Number of animals H&E staining score Control 8 0.41±0.19 pH 4.0 8 1.37±0.24* pH 4.5 8 0.75±0.35 pH 5.0 8 0.47±0.26 pH 5.5 8 0.45±0.21 pH 6.0 8 0.46±0.23 pH 6.5 8 0.42±0.17 pH 7.0 8 0.39±0.20 pH 7.5 8 0.65±0.29 pH 8.0 8 0.79±0.25

[0119] Statistics: x±SEM, n=8, one way ANOVA, compared with the Control group: *p<0.05.

[0120] Table 7 shows that when pH=4, repeated administration for 30 days, twice a day, there is a certain irritation, p<0.05; when pH value is ≦4.5 or ≧7.5, long-term administration of guinea pigs has a tendency to irritate the nasal mucosa, but the P value is greater than 0.05; when the pH is between 5-7, long-term administration of guinea pigs has no obvious irritation or tendency to the nasal mucosa.

[0121] ②The influence of NaCl content: the NaCl content of the test drug nasal spray was 0%, 0.45%, 0.9%, 1.35%, 1.8%, 2.25% and 2.70%, and other components were the same as in Example 2, the irritation of different NaCl content to the nasal mucosa of guinea pigs is shown in Table 8:

[0122] Table 8, the irritation of different NaCl concentration to the nasal mucosa of guinea pigs (x±SEM)

[0123] Group Number of animals H&E staining score Control 8 0.41±0.19 NaCl (0%) 8 0.85±0.31 NaCl (0.45%) 8 0.47±0.24 NaCl (0.90%) 8 0.40±0.15 NaCl (1.35%) 8 0.49±0.23 NaCl (1.80%) 8 0.68±0.32 NaCl (2.25%) 8 0.92±0.25 NaCl (2.70%) 8 1.67±0.33*

[0124] Statistics: x±SEM, n=8, one way ANOVA, compared with the Control group: *p<0.05.

[0125] Table 8 shows that when the NaCl content deviates from 0.90% to 2.70%, repeated administration for 30 days, twice a day, has a significant stimulating effect, p<0.05; the greater the deviation of the NaCl content from 0.9%, the more obvious the trend of the stimulating effect of long-term administration of guinea pigs on the nasal mucosa; the stimulating effect of 0.45%≦NaCl content≦2.25% on the nasal mucosa has no significant difference compared with the control group (p>0.05).

[0126] 3. Effect of cosolvents: The cosolvents of the test drug nasal spray are selected as ethanol (content of 5%, 10%, 15%), ethylene glycol 200 (content of 5%, 10%, 15%) and a mixture of ethylene glycol 200 and ethanol 1:1 (content of 5%, 10%, 15%); the main drug is compound 1, with a content of 0.5%; other ingredients are the same as in Example 2. The stimulating effect of different contents of cosolvents on the nasal mucosa of guinea pigs is shown in Table 9:

[0127] Table 9, stimulating effect of different contents of cosolvents on the nasal mucosa of guinea pigs (x±SEM)

[0128] Group Number of animals H&E staining score Control 8 0.41±0.19 Ethanol (5%) 8 0.53±0.27 Ethanol (10%) 8 0.76±0.32 Ethanol (15%) 8 1.58±0.31* Ethylene glycol (5%) 8 0.45±0.20 Ethylene glycol (10%) 8 0.63±0.28 Ethylene glycol (15%) 8 1.51±0.27* Mixture (5%) 8 0.458±0.23 Mixture (10%) 8 0.67±0.24 Mixture (15%) 8 1.54±0.25*

[0129] Statistics: x±SEM, n=8, one way ANOVA, compared with the Control group: *p<0.05.

[0130] Table 9 shows that cosolvents at a concentration less than 5% do not have a significant stimulating effect; as the concentration of cosolvents increases, the stimulating effect tends to increase; when the content of various cosolvents exceeds 10%, repeated administration for 30 days, twice a day, has a significant stimulating effect, and p<0.05.

Claims

1. A nasal spray containing a compound of formula I, water, a cosolvent, an antibacterial agent, a pH adjuster, and NaCl, wherein the pH is 6-7, the content of the compound of formula I is 0.02-0.5% by weight, the content of the cosolvent ethanol is 0-5% by weight, the content of the antibacterial agent is 0.005-0.02% by weight, and the content of NaCl is 0.45-1.35% by weight. M - It is a bromide ion.

2. The nasal spray according to claim 1, wherein the antibacterial agent is benzalkonium chloride or benzalkonium bromide.

3. The nasal spray according to any one of claims 1-2, wherein the pH adjuster is selected from one or more of HCl, H3PO4, H2SO4, lactic acid, citric acid, tartaric acid, malic acid, maleic acid, succinic acid, benzoic acid, sodium salts of the aforementioned acids other than HCl, and NaOH.

4. The method for preparing a nasal spray according to any one of claims 1-3, wherein the compound of formula I is dissolved in water or dissolved in a co-solvent and then dissolved in water, an antibacterial agent and an osmotic pressure regulator NaCl are added to dissolve the compound, the pH is adjusted, and then it is filled into a container.

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