A lysozyme nasal spray and preparation method thereof

Abstract

The present application discloses a lysozyme nasal spray, and its raw material composition includes, by weight: 45-53 parts of lysozyme, 65-75 parts of viscosity modifier, 85-90 parts of sodium chloride, 2.5-3.5 parts of magnesium chloride, 0.7-0.8 parts of benzalkonium chloride, 1920-2030 parts of poloxamer 407, and 7950-8035 parts of water for injection. The present application also provides a method for preparing a lysozyme nasal spray, comprising the following steps: S1, adding the lysozyme, viscosity modifier, sodium chloride, magnesium chloride, benzalkonium chloride and poloxamer 407 to water for injection, and then stirring to obtain a mixed solution; S2, adjusting the pH of the mixed solution to 6.5-7.5, and then performing sterilization filtration, and the filtrate is the lysozyme nasal spray. The present application adopts specific proportions of components, dosages and pH values, and the prepared nasal spray is mild and non-irritating, has a broad-spectrum antibacterial effect, and is easy to use.

Description

Technical Field

[0001] The present application relates to the technical field of pharmaceutical preparations, and in particular to a lysozyme nasal spray and a preparation method thereof. Background Art

[0002] Viruses or pathogenic microorganisms such as coronavirus, rhinovirus, influenza virus and other pathogens can easily cause diseases after invading the human body from the nasal cavity, throat, trachea and other parts. Existing antiviral preparations are taken by brewing, injection or nasal administration. Brewing and swallowing requires 1 to 2 hours of body metabolism before it can play an antibacterial and antiviral role. Injection administration is not convenient for self-medication, and children have poor compliance and are not easily accepted by children, which is inconvenient to use.

[0003] Nasal administration is convenient and has good patient compliance. The surface area of ​​the human nasal mucosa is 150 cm 2 There are many microvilli on the epithelial cells on the mucosal surface, which are similar to the villi in the small intestine, which can increase the effective area for drug absorption. In addition, there are abundant capillaries and capillary lymphatic vessels in the subepithelial layer of the nasal mucosa, which can quickly absorb drugs into the human circulation. The mucosa in the upper nasal cavity is thicker than that in the bottom of the nasal cavity and the sinuses, and has dense blood vessels. It is the main area for drug absorption. The nasal mucosa is also considered to be the first line of defense against antigen invasion. Therefore, nasal administration has advantages for diseases infected through the respiratory tract. In addition, nasal administration of drugs does not pass through the gastrointestinal tract but directly reaches the site of action. A small amount of drug can achieve a higher blood concentration, generally only about 1 / 10 or 1 / 5 of the oral drug dose is required, thereby reducing the toxic and side effects of the drug.

[0004] Therefore, nasal administration avoids the first-pass effect of oral administration and the destructive effects of the gastrointestinal tract. Patients can administer the drug in a quantitative manner by themselves, which is convenient to use and has good patient compliance. However, current nasal administration agents are prone to irritation to the nasal mucosa and have a narrow antibacterial spectrum. Therefore, if nasal administration agents are mild to the nasal mucosa and have a broad spectrum of antibacterial, it will bring good news to patients. Summary of the invention

[0005] In order to solve at least one of the above technical problems, a nasal spray that is mild and non-irritating to the nasal mucosa and has a broad-spectrum antibacterial effect is developed. The present application provides a lysozyme nasal spray and a preparation method thereof.

[0006] On the one hand, the present application provides a lysozyme nasal spray, the raw material composition of which includes, by weight:

[0007]

[0008]

[0009] By adopting the above technical scheme, the nasal spray prepared by the application using specific components and dosage is mild and non-irritating, avoiding the nasal spray from irritating the nasal mucosa, and also has a broad-spectrum antibacterial and antiviral effect, and is easy to use and has good patient compliance. Lysozyme has a wide range of antibacterial, antiviral, and immune-enhancing effects, and is easily absorbed, easily digested, has good biocompatibility, is non-irritating to tissues, non-toxic, and does not have drug residue problems; the viscosity modifier is used to adjust the initial viscosity of the spray so that the spray can be sprayed out of the spray device more smoothly, and will not flow out immediately after entering the nasal cavity; after the spray enters the nasal cavity and falls on the nasal mucosa, since the temperature of the nasal mucosa is about 36.5°C, the temperature of the spray in contact with the nasal mucosa increases, and the temperature of poloxamer 407 in the spray also increases, thereby changing the spray from a liquid state to a gel state, thereby extending the time the agent stays in the nasal cavity, and thereby enabling the agent to better play an antibacterial and antiviral role. The nasal spray prepared in the present application is stored below 30° C. to prevent the spray from forming gel in the spray device and causing it to be unable to spray during use.

[0010] Optionally, the raw material composition includes, by weight:

[0011]

[0012] Optionally, the lysozyme is hen egg white lysozyme.

[0013] Optionally, the viscosity modifier is hypromellose.

[0014] Optionally, the pH value of the lysozyme nasal spray is 6.5-7.5.

[0015] Optionally, the viscosity of the lysozyme nasal spray is 6 to 8 mPa·s.

[0016] By adopting the above technical solution, the nasal spray maintains an appropriate viscosity, can be sprayed out from the spray device more smoothly, and will not flow out immediately after entering the nasal cavity, which helps to improve the efficacy of the medicine.

[0017] In a second aspect, the present application provides a method for preparing the above-mentioned lysozyme nasal spray, comprising the following steps:

[0018] S1, adding the lysozyme, the viscosity modifier, the sodium chloride, the magnesium chloride, the benzalkonium chloride and the poloxamer 407 into the water for injection, and then stirring to obtain a mixed solution;

[0019] S2. The pH of the mixed solution is adjusted to 6.5-7.5, and then sterilized and filtered. The filtrate is the lysozyme nasal spray.

[0020] By adopting the above technical solution, the present application adopts specific proportions of components, dosages and pH, and the prepared nasal spray is mild and non-irritating, and has a broad-spectrum antibacterial and antiviral effect.

[0021] Optionally, in step S2, a polyethersulfone sterilizing filter is used for sterilizing filtration treatment, and the pore size of the filter pores of the polyethersulfone sterilizing filter is 0.22 um.

[0022] In summary, the present application includes at least one of the following beneficial technical effects:

[0023] 1. The present application adopts specific proportions of components, dosages and pH values, and the prepared nasal spray is mild and non-irritating, has broad-spectrum antibacterial and antiviral effects, and is easy to use.

[0024] 2. Lysozyme has a wide range of antibacterial, antiviral, and immunity-enhancing effects. It is easily absorbed, easily digested, has good biocompatibility, is non-irritating and non-toxic to tissues, and does not cause drug residue problems.

[0025] 3. The viscosity modifier is used to adjust the initial viscosity of the spray so that the spray can be sprayed out of the spray device more smoothly and will not flow out immediately after entering the nasal cavity.

[0026] 4. After the spray enters the nasal cavity and falls on the nasal mucosa, the temperature of the nasal mucosa is about 36.5°C, which increases the temperature of the spray in contact with the nasal mucosa. The temperature of the poloxamer 407 in the spray also increases, causing the spray to change from liquid to gel, thereby prolonging the time the drug stays in the nasal cavity, allowing the drug to better play an antibacterial and antiviral role. DETAILED DESCRIPTION

[0027] The present application is further described in detail below with reference to the embodiments.

[0028] The present application designs a lysozyme nasal spray, and its raw material composition includes, by weight:

[0029]

[0030] The lysozyme nasal spray of the present application is prepared by the following method, comprising the following steps:

[0031] S1, adding the lysozyme, the viscosity modifier, the sodium chloride, the magnesium chloride, the benzalkonium chloride and the poloxamer 407 into the water for injection, and then stirring to obtain a mixed solution;

[0032] S2. The pH of the mixed solution is adjusted to 6.5-7.5, and then sterilized and filtered. The filtrate is the lysozyme nasal spray.

[0033] The lysozyme nasal spray of the present application can be used in the field of fighting coronavirus, rhinovirus and influenza virus.

[0034] The present application adopts a specific proportion of components, dosage and pH, and the prepared nasal spray is mild and non-irritating, has no irritating adverse reactions to the nasal mucosa, and also has broad-spectrum antibacterial and antiviral effects, and is easy to use and has good patient compliance. Lysozyme has a wide range of antibacterial, antiviral, and immune-enhancing effects, and is easily absorbed, easily digested, has good biocompatibility, is non-irritating and non-toxic to tissues, and does not have drug residue problems; the viscosity modifier is used to adjust the initial viscosity of the spray, so that the spray can be sprayed out of the spray device more smoothly, and will not flow out immediately after entering the nasal cavity; poloxamer 407 changes the spray from a liquid state to a gel state after entering the nasal cavity, thereby prolonging the time the agent stays in the nasal cavity, and then making the agent better play the role of antibacterial and antiviral. Specific embodiments

[0036] Example 1

[0037] A lysozyme nasal spray comprises the following components and the proportions of the components: 50 g lysozyme, 70 g hydroxypropyl methylcellulose, 87 g sodium chloride, 3 g magnesium chloride, 0.75 g benzalkonium chloride, 2000 g poloxamer 407 and 8000 ml water for injection.

[0038] The lysozyme is the hen egg white lysozyme from Shanghai Zhonghua Pharmaceutical Co., Ltd., the hydroxypropyl methylcellulose is purchased from Anhui Shanhe Pharmaceutical Excipients Co., Ltd., the benzalkonium chloride is purchased from Hubei Gedian Renfu Pharmaceutical Excipients Co., Ltd., and the poloxamer 407 is purchased from Shanghai Yunhong Chemical Preparation Excipients Technology Co., Ltd.

[0039] A method for preparing a lysozyme nasal spray comprises the following steps:

[0040] S1, adding the lysozyme, the viscosity modifier, the sodium chloride, the magnesium chloride, the benzalkonium chloride and the poloxamer 407 into the water for injection, and then stirring to obtain a mixed solution;

[0041] S2. The pH of the mixed solution is adjusted by using hydrochloric acid and sodium hydroxide to make the pH of the mixed solution 6.5, and then the mixed solution is sterilized and filtered by using a polyethersulfone sterilizing filter, wherein the pore size of the filter hole of the polyethersulfone sterilizing filter is 0.22um, and the filtrate is the lysozyme nasal spray, and the lysozyme nasal spray is filled into polyethylene spray bottles, each bottle containing 10ml of the lysozyme nasal spray. The concentration of hydrochloric acid is 1mol / L.

[0042] When using, aim the spray head of the spray bottle at the nasal cavity and press the spray head twice or three times to make the lysozyme nasal spray more evenly distributed in the nasal cavity, thereby playing an antibacterial and antiviral role.

[0043] Example 2

[0044] A lysozyme nasal spray comprises the following components and the proportions of the components: 50 g lysozyme, 65 g hydroxypropyl methylcellulose, 85.5 g sodium chloride, 3.1 g magnesium chloride, 0.72 g benzalkonium chloride, 2040 g poloxamer 407 and 8030 ml water for injection.

[0045] A method for preparing a lysozyme nasal spray comprises the following steps:

[0046] S1, adding the lysozyme, the viscosity modifier, the sodium chloride, the magnesium chloride, the benzalkonium chloride and the poloxamer 407 into the water for injection, and then stirring to obtain a mixed solution;

[0047] S2. The pH of the mixed solution is adjusted with hydrochloric acid and sodium hydroxide to a pH of 6.5. Then, a polyethersulfone sterilizing filter is used for sterilizing filtration. The pore size of the filter pores of the polyethersulfone sterilizing filter is 0.22 um. The filtrate is the lysozyme nasal spray.

[0048] Example 3

[0049] A lysozyme nasal spray comprises the following components and the proportions of the components: 45 g lysozyme, 70 g hydroxypropyl methylcellulose, 90 g sodium chloride, 2.5 g magnesium chloride, 0.7 g benzalkonium chloride, 2000 g poloxamer 407 and 8035 ml water for injection.

[0050] A method for preparing a lysozyme nasal spray comprises the following steps:

[0051] S1, adding the lysozyme, the viscosity modifier, the sodium chloride, the magnesium chloride, the benzalkonium chloride and the poloxamer 407 into the water for injection, and then stirring to obtain a mixed solution;

[0052] S2. The pH of the mixed solution is adjusted with hydrochloric acid and sodium hydroxide to be 7, and then a polyethersulfone sterilizing filter is used for sterilizing filtration. The pore size of the filter pores of the polyethersulfone sterilizing filter is 0.22 um, and the filtrate is the lysozyme nasal spray.

[0053] Example 4

[0054] A lysozyme nasal spray comprises the following components and the proportions of the components: 53g lysozyme, 70.5g hydroxypropyl methylcellulose, 87g sodium chloride, 3.5g magnesium chloride, 0.7g benzalkonium chloride, 1920g poloxamer 407 and 7950ml water for injection.

[0055] A method for preparing a lysozyme nasal spray comprises the following steps:

[0056] S1, adding the lysozyme, the viscosity modifier, the sodium chloride, the magnesium chloride, the benzalkonium chloride and the poloxamer 407 into the water for injection, and then stirring to obtain a mixed solution;

[0057] S2. The pH of the mixed solution is adjusted with hydrochloric acid and sodium hydroxide to be 7, and then a polyethersulfone sterilizing filter is used for sterilizing filtration. The pore size of the filter pores of the polyethersulfone sterilizing filter is 0.22 um, and the filtrate is the lysozyme nasal spray.

[0058] Example 5

[0059] A lysozyme nasal spray comprises the following components and the proportions of the components: 50 g lysozyme, 75 g hydroxypropyl methylcellulose, 85 g sodium chloride, 3 g magnesium chloride, 0.74 g benzalkonium chloride, 2030 g poloxamer 407 and 8000 ml water for injection.

[0060] A method for preparing a lysozyme nasal spray comprises the following steps:

[0061] S1, adding the lysozyme, the viscosity modifier, the sodium chloride, the magnesium chloride, the benzalkonium chloride and the poloxamer 407 into the water for injection, and then stirring to obtain a mixed solution;

[0062] S2. The pH of the mixed solution is adjusted with hydrochloric acid and sodium hydroxide to a pH of 7.5. After that, a polyethersulfone sterilizing filter is used for sterilizing filtration. The pore size of the filter pores of the polyethersulfone sterilizing filter is 0.22 um. The filtrate is the lysozyme nasal spray.

[0063] Example 6

[0064] A lysozyme nasal spray comprises the following components and the proportions of the components: 52.5 g lysozyme, 68.5 g hydroxypropyl methylcellulose, 86 g sodium chloride, 2.8 g magnesium chloride, 0.8 g benzalkonium chloride, 1980 g poloxamer 407 and 8020 ml water for injection.

[0065] A method for preparing a lysozyme nasal spray comprises the following steps:

[0066] S1, adding the lysozyme, the viscosity modifier, the sodium chloride, the magnesium chloride, the benzalkonium chloride and the poloxamer 407 into the water for injection, and then stirring to obtain a mixed solution;

[0067] S2. The pH of the mixed solution is adjusted with hydrochloric acid and sodium hydroxide to a pH of 7.5. After that, a polyethersulfone sterilizing filter is used for sterilizing filtration. The pore size of the filter pores of the polyethersulfone sterilizing filter is 0.22 um. The filtrate is the lysozyme nasal spray.

[0068] Example 7

[0069] A lysozyme nasal spray comprises the following components and the proportions of the components: 48 g lysozyme, 70 g hydroxypropyl methylcellulose, 87 g sodium chloride, 2.9 g magnesium chloride, 0.73 g benzalkonium chloride, 2000 g poloxamer 407 and 8015 ml water for injection.

[0070] A method for preparing a lysozyme nasal spray comprises the following steps:

[0071] S1, adding the lysozyme, the viscosity modifier, the sodium chloride, the magnesium chloride, the benzalkonium chloride and the poloxamer 407 into the water for injection, and then stirring to obtain a mixed solution;

[0072] S2. The pH of the mixed solution is adjusted with hydrochloric acid and sodium hydroxide to a pH of 7.5. After that, a polyethersulfone sterilizing filter is used for sterilizing filtration. The pore size of the filter pores of the polyethersulfone sterilizing filter is 0.22 um. The filtrate is the lysozyme nasal spray.

[0073] Example 8

[0074] A lysozyme nasal spray comprises the following components and the proportions of the components: 50 g lysozyme, 67 g hydroxypropyl methylcellulose, 87.5 g sodium chloride, 3.2 g magnesium chloride, 0.73 g benzalkonium chloride, 1970 g poloxamer 407 and 8000 ml water for injection.

[0075] A method for preparing a lysozyme nasal spray comprises the following steps:

[0076] S1, adding the lysozyme, the viscosity modifier, the sodium chloride, the magnesium chloride, the benzalkonium chloride and the poloxamer 407 into the water for injection, and then stirring to obtain a mixed solution;

[0077] S2. The pH of the mixed solution is adjusted with hydrochloric acid and sodium hydroxide to a pH of 6.5. Then, a polyethersulfone sterilizing filter is used for sterilizing filtration. The pore size of the filter pores of the polyethersulfone sterilizing filter is 0.22 um. The filtrate is the lysozyme nasal spray.

[0078] Comparative Example 1

[0079] The difference between Comparative Example 1 and Example 1 is that lysozyme is replaced by ganciclovir, and the ganciclovir is purchased from Shanghai En Chemical Technology Co., Ltd.

[0080] Irritation test

[0081] The sprays prepared in Examples 1 to 8 and Comparative Example 1 were subjected to a nasal mucosal irritation test.

[0082] Fifty-four rats were randomly divided into nine groups, each with six rats, eight of which were example groups and one group was a control group. A soft PE tube was connected in front of the syringe, and one end of the soft PE tube was inserted into the left nasal cavity of the rat. The spray (10 mg / kg) prepared in Examples 1 to 8 was slowly introduced into the example group, and the spray (10 mg / kg) prepared in Comparative Example 1 was slowly introduced into the control group. After 24 hours, two rats in each group were randomly killed, and then the left nasal cavity was cut open to observe the rat's nasal mucosa, and the spray (10 mg / kg) was pushed into the left nasal cavity of the corresponding remaining rat. After 48 hours, two rats in each group were randomly killed, and then the left nasal cavity was cut open to observe the rat's nasal mucosa, and the spray (10 mg / kg) was pushed into the left nasal cavity of the corresponding remaining rat. After 72 hours, the remaining rats in each group were killed, and then the left nasal cavity was cut open to observe the rat's nasal mucosa. The observation results are shown in Table 1 below.

[0083] Table 1

[0084]

[0085] “+” indicates that the rat’s nasal mucosa is congested or swollen, and “—” indicates that the rat’s nasal mucosa is not congested or swollen.

[0086] It can be seen from the test results in Table 1 that the lysozyme nasal spray prepared in the present application is mild and non-irritating to the nasal mucosa.

[0087] Comparative Example 2

[0088] The difference between Comparative Example 2 and Example 1 is that the content of lysozyme is 43 g.

[0089] Antibacterial test

[0090] The sprays prepared in Examples 1 to 8 and Comparative Example 2 were tested according to GB15979-2002 "Hygiene Standard for Disposable Hygiene Products". The prepared sprays were used to act on Staphylococcus aureus, Bacillus subtilis, Streptococcus mutans, Escherichia coli, Pseudomonas aeruginosa, and Candida albicans, respectively. The action time on each bacterium was 2 minutes. The test was repeated 3 times at 25±2°C, and the antibacterial rate was the average value of the 3 times. The test results are shown in Table 2.

[0091] Table 2

[0092] From the test results in Table 2, it can be seen that the lysozyme nasal spray prepared in the present application has a wide range of antibacterial properties, and the antibacterial rate reaches more than 95%.

[0093] Comparative Example 3

[0094] The difference between Comparative Example 3 and Example 1 is that the content of the hypromellose is 60 g.

[0095] The difference between Comparative Example 4 and Example 1 is that the content of the hypromellose is 80 g.

[0096] Initial viscosity test

[0097] 8 ml of the sprays prepared in Examples 1 to 8 and Comparative Examples 3 and 4 were taken respectively, and the initial viscosity test was performed using an LVDV-27 rotational viscometer with a No. 18 rotor and a rotation speed of 150 rpm at a temperature of 25°C. The test was performed 3 times, and the initial viscosity value was the average value of the 3 times. The test results are shown in Table 3 below.

[0098] Table 3

[0099] Initial viscosity (mPa·s) Example 1 6.00 Example 2 6.21 Example 3 7.23 Example 4 6.52 Example 5 6.21 Example 6 8.00 Example 7 6.23 Example 8 6.23 Comparative Example 3 5.14 Comparative Example 4 8.95

[0100] From the test results in Table 3, it can be seen that in Comparative Example 3, the content of hypromellose is lower than 65g, and the initial viscosity value of the prepared spray is low. Although the spray can be sprayed out of the spray bottle more smoothly, the spray is easy to flow out after being sprayed into the nasal cavity; the content of hypromellose in Comparative Example 4 is higher than 75g, and the initial viscosity value of the prepared spray is higher, and the spray cannot be sprayed out of the spray bottle more smoothly. The content of hypromellose in the present application is 65-75g, which can make the spray spray out of the spray bottle more smoothly, and the spray will not flow out immediately after being sprayed into the nasal cavity.

[0101] Comparative Example 5

[0102] The difference between Comparative Example 5 and Example 1 is that Comparative Example 5 does not contain Poloxamer 407.

[0103] Viscosity change test with temperature

[0104] Take 9 test tubes, add the sprays prepared in Examples 1 to 8 and Comparative Example 5 respectively, then place the test tubes in an environment of 34°C for 2 minutes, invert the test tubes, and observe the flow of the liquid within 0.5 minutes. The test results are shown in Table 4 below.

[0105] Table 4

[0106]

[0107]

[0108] From the test results in Table 4, it can be seen that Examples 1 to 8 contain a specific proportion of Poloxamer 407, which can make the spray change from liquid to gel at 34°C, so that no flow will occur within 0.5 minutes. However, Comparative Example 5 does not contain Poloxamer 407, so it will flow when the test tube is inverted.

[0109] The above are all preferred embodiments of the present application, and the protection scope of the present application is not limited thereto. Therefore, any equivalent changes made according to the structure, shape, and principle of the present application should be included in the protection scope of the present application.

Claims

1. A lysozyme nasal spray, characterized in that: The raw material composition is as follows by weight: 45-53 parts of lysozyme, 65-75 parts of viscosity modifier, 85-90 parts of sodium chloride, 2.5-3.5 parts of magnesium chloride, Benzalkonium chloride 0.7-0.8 parts, Poloxamer 407 1920~2030 parts, Water for injection 7950-8035 parts; The viscosity modifier is hypromellose; The viscosity of the lysozyme nasal spray is 6 to 8 mPa•s; The pH value of the lysozyme nasal spray is 6.5-7.

5.

2. The lysozyme nasal spray according to claim 1, characterized in that The raw material composition is as follows by weight: Lysozyme 50 parts, Viscosity adjustment 70 parts, Sodium chloride 87 parts, 3 parts of magnesium chloride, 0.75 parts of benzalkonium chloride, Poloxamer 407 2000 copies, 8000 parts of water for injection.

3. The lysozyme nasal spray according to claim 1, characterized in that The lysozyme is hen egg white lysozyme.

4. A method for preparing the lysozyme nasal spray according to claim 1, characterized in that: The steps include: S1, adding the lysozyme, the viscosity modifier, the sodium chloride, the magnesium chloride, the benzalkonium chloride and the poloxamer 407 into the water for injection, and then stirring to obtain a mixed solution; S2. The pH of the mixed solution is adjusted to 6.5-7.5, and then sterilized and filtered. The filtrate is the lysozyme nasal spray.

5. The method for preparing the lysozyme nasal spray according to claim 4, characterized in that: In step S2, a polyethersulfone sterilizing filter is used for sterilizing filtration treatment, and the pore size of the filter pores of the polyethersulfone sterilizing filter is 0.22 um.

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