A composition containing oseltamivir phosphate and its use in preparing antiviral drugs

Abstract

The present invention belongs to the technical field of antiviral drug preparations, and specifically relates to a composition containing oseltamivir phosphate and its use in preparing antiviral drugs. The composition containing oseltamivir phosphate of the present invention is composed of oseltamivir phosphate, pulegone, soybean lecithin or / and egg yolk lecithin, and cholesterol. Oseltamivir phosphate and pulegone are encapsulated by liposome technology, and the structural integrity of the liposome and the stability of the drug are maintained by dual regulation of glucose and pH, and the content of related substances is reduced. Animal experiments are also used to carry out drug efficacy verification, and the therapeutic effect is better than that of commercially available preparations.

Description

Technical Field

[0001] The present invention belongs to the technical field of antiviral drug preparations, and in particular relates to a composition containing oseltamivir phosphate and use thereof in preparing antiviral drugs. Background Art

[0002] The information disclosed in this background technology section is only intended to enhance the understanding of the overall background of the invention, and should not necessarily be regarded as an admission or any form of suggestion that the information constitutes the prior art already known to a person skilled in the art.

[0003] Influenza is an infectious respiratory disease caused by influenza viruses, which is common in winter and spring. Influenza A and B are the two main types of influenza viruses, which cause most influenza cases during the flu season. Influenza A viruses are mainly transmitted by birds and mammals, including humans. Influenza A viruses are divided into different subtypes, based on changes in two proteins on their surface (hemagglutinin and neuraminidase), the most common subtypes are H1N1 and H3N2. Influenza B viruses are mainly transmitted by humans, and a few can also be transmitted between animals. Influenza B mutates less, so it does not change significantly as often as influenza A. Influenza B does not usually cause a global pandemic compared to influenza A, but can still cause a seasonal disease burden.

[0004] Whether it is influenza A or influenza B, they all have the following common characteristics: (1) Symptoms: Common symptoms of influenza include sudden fever, headache, muscle pain, cough, sore throat, fatigue and chills. (2) Mode of transmission: Influenza is mainly transmitted through airborne droplets, and can also be transmitted through contact with contaminated surfaces.

[0005] Oseltamivir phosphate was originally developed by Swiss pharmaceutical company Roche and approved for marketing in the United States in 1999. It is a drug for the treatment of influenza virus infection and is effective in treating influenza A and B. Oseltamivir phosphate is a neuraminidase inhibitor that blocks the replication and reproduction of the virus in host cells by inhibiting the neuraminidase activity of the influenza virus. It mainly targets the neuraminidase of the influenza virus, thereby reducing viral replication, shortening the duration of the disease, and reducing the spread of the virus in the host.

[0006] Chinese patent publication number CN116869939A discloses an oseltamivir phosphate liposome, a preparation method and a preparation. By using liposome technology, oseltamivir phosphate is encapsulated by egg yolk phospholipids, stigmasterol, 2-hydroxyethylamine and sodium citrate in a certain proportion, providing a high-quality oseltamivir phosphate liposome and an oseltamivir acid preparation with low content of related substances and high stability.

[0007] Chinese patent publication number CN108420792A discloses a pharmaceutical composition containing oseltamivir phosphate, which is encapsulated in the inner aqueous phase of liposomes by active drug loading using ammonium sulfate gradient, effectively avoiding direct contact with the oral cavity and achieving a good taste masking effect. The oseltamivir phosphate liposome suspension is made into a dry powder by spray drying technology. Summary of the invention

[0008] The invention provides an oseltamivir phosphate composition with high stability and good antiviral effect. Specifically, the oseltamivir phosphate composition comprises 10-40 parts by weight of oseltamivir phosphate, 1-5 parts by weight of pulegone (CAS: 89-80-5, purity ≥98%), 15-50 parts by weight of soybean lecithin or / and egg yolk lecithin, and 5-20 parts by weight of cholesterol. The preparation method of the composition comprises the following steps: dissolving pulegone, soybean lecithin or / and egg yolk lecithin, and cholesterol in an organic solvent, placing the mixture in an eggplant-shaped bottle, heating in a water bath and performing vacuum rotary evaporation to remove the organic solvent, adding a glucose-buffer solution mixture containing oseltamivir phosphate to hydrate the mixture, continuing heating in a water bath and performing vacuum rotary evaporation, performing high pressure homogenization, and performing freeze drying to obtain the oseltamivir phosphate composition.

[0009] Furthermore, the glucose-buffer solution mixture is a glucose-phosphate buffer solution with a mass fraction of 4.5% to 9.5%. The addition of an appropriate amount of glucose can reduce the crystallization of liposomes and the movement of lipid molecules, thereby maintaining the structural integrity of the liposomes and the stability of the drug. In addition, glucose can also reduce the dehydration rate of liposomes during freeze-drying through osmotic protection, thereby reducing the formation of crystals and the destruction of liposomes.

[0010] Furthermore, the organic solvent is selected from one or more of methanol, ethanol, and n-butanol, preferably, a methanol-ethanol mixed solvent with a volume ratio of 3 to 5:1, and the organic solvents are all analytical grade with a purity of ≥95%.

[0011] Furthermore, the pH of the glucose-buffered solution is 7.5 to 8.5, and the purpose is to improve the stability of the oseltamivir phosphate liposome composition, reduce leakage, and reduce the content of related substances by combining with the glucose solution.

[0012] Furthermore, in order to remove the organic solvent, the water bath temperature is 30° C. to 50° C., and can be appropriately adjusted within the temperature range according to the outflow rate of the distillate.

[0013] For the convenience of taking medicine, the present invention also provides a preparation consisting of the above-mentioned composition containing oseltamivir phosphate and pharmaceutically acceptable excipients. The preparation can be one or more of tablets, granules, capsules, pills, and oral liquids. The pharmaceutically acceptable excipients are selected from at least one of fillers, lubricants, disintegrants, binders, humectants, solvents, flavoring agents, preservatives, stabilizers, thickeners, and surfactants. The types and amounts of the excipients are conventional weights and amounts, and are recorded in relevant textbooks in the field.

[0014] In order to prove the synergistic effect of oseltamivir phosphate and pulegone, the present invention also uses animal experiments to verify the use of the above-mentioned composition containing oseltamivir phosphate in the preparation of antiviral drugs.

[0015] Compared with the prior art, the technical effects of the present invention are:

[0016] The present invention utilizes liposome technology to encapsulate oseltamivir phosphate and pulegone. Oseltamivir phosphate as a water-soluble drug is encapsulated in an aqueous phase medium of a polar layer, and pulegone as an oil-soluble drug is encapsulated in a hydrophobic chain layer to form a closed double-layer structure. The structural integrity of the liposome and the stability of the drug are maintained through dual regulation of glucose and pH, and the content of related substances is reduced.

[0017] The present invention uses animal experiments to verify the efficacy and finds that oseltamivir phosphate and pulegone do have a synergistic effect, and the therapeutic effect is better than that of commercially available preparations. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 : Electron microscopic image of the composition containing oseltamivir phosphate in Example 1.

[0019] Figure 2 : Comparison of drug loading of the compositions containing oseltamivir phosphate in Examples 1 to 3 and the compositions containing oseltamivir phosphate in Comparative Examples 1 to 4.

[0020] Figure 3 : The encapsulation efficiency and leakage rate of the compositions containing oseltamivir phosphate in Examples 1 to 3 and the compositions containing oseltamivir phosphate in Comparative Examples 1 to 4.

[0021] Figure 4 : Effect of glucose concentration on the leakage rate of oseltamivir phosphate compositions.

[0022] Figure 5 : Effect of pH on the leakage rate of oseltamivir phosphate compositions.

[0023] Figure 6 :Impurity Ⅰ(3R,4R,5S)-4-acetylamino-5-amino-3-(1-ethylpropoxy)-1-cyclohexene-1-carboxylic acid methyl ester phosphate.

[0024] Figure 7 : Impurity ⅠⅠ 3-hydroxy-4-acetylaminobenzoic acid ethyl ester.

[0025] Figure 8 : Impurity ⅠⅠⅠ(3R,4R,5S)-4-acetylamino-5-amino-3-(1-ethylpropoxy)-1-cyclohexene-1-carboxylic acid.

[0026] Fig. 9 : Changes in the contents of related substances in the oseltamivir phosphate preparations of Examples 4 to 9 and the oseltamivir phosphate capsules of Comparative Example 6.

[0027] Fig.10 : Body temperature of rats in each group after administration. DETAILED DESCRIPTION

[0028] In order to make the purpose and technical solution of the present invention clearer, the present invention is further described below in conjunction with embodiments, but the protection scope of the present invention is not limited to these embodiments, and the embodiments are only used to explain the present invention. It should be understood by those skilled in the art that any changes or equivalent substitutions that do not deviate from the concept of the present invention are included in the protection scope of the present invention.

[0029] Example 1 Composition of Oseltamivir Phosphate

[0030] prescription:

[0031]

[0032]

[0033] Preparation method:

[0034] (1) Pulegone, soybean lecithin, egg yolk lecithin, and cholesterol were dissolved in a methanol-ethanol mixed solvent with a volume ratio of 4:1, placed in an eggplant-shaped bottle, heated in a water bath at 35°C and evaporated under reduced pressure to remove the solvent;

[0035] (2) After the film is formed, a glucose-phosphate buffer solution mixture (pH=8.0) containing 6.5% by mass of oseltamivir phosphate is added for hydration. After sufficient hydration, the mixture is heated in a water bath at 45° C. and subjected to reduced pressure rotary evaporation, high pressure homogenization, and freeze-dried to obtain a composition containing oseltamivir phosphate.

[0036] Example 2 Composition of Oseltamivir Phosphate

[0037] prescription:

[0038] Dosage(g) Oseltamivir phosphate 10 Pulegone 1 Soybean lecithin 15 cholesterol 5

[0039] Preparation method:

[0040] (1) Pulegone, soybean lecithin, and cholesterol were dissolved in a methanol-ethanol mixed solvent with a volume ratio of 3:1, placed in an eggplant-shaped bottle, heated in a water bath at 30°C and evaporated under reduced pressure to remove the solvent;

[0041] (2) After the film is formed, a mixed solution of glucose-phosphate buffer solution (pH=7.5) containing 4.5% by mass of oseltamivir phosphate is added for hydration. After sufficient hydration, the mixture is heated in a water bath at 50° C. and subjected to reduced pressure rotary evaporation, high pressure homogenization, and freeze-dried to obtain a composition containing oseltamivir phosphate.

[0042] Example 3 Composition of Oseltamivir Phosphate

[0043] prescription:

[0044] Dosage(g) Oseltamivir phosphate 40 Pulegone 5 Egg yolk lecithin 50 cholesterol 20

[0045] Preparation method:

[0046] (1) dissolving pulegone, soybean lecithin or / and egg yolk lecithin, and cholesterol in a methanol-ethanol mixed solvent with a volume ratio of 5:1, placing the mixture in an eggplant-shaped bottle, heating in a water bath at 35°C and vacuum evaporating the mixture to remove the solvent;

[0047] (2) After the film is formed, a glucose-phosphate buffer solution mixture (pH=8.5) containing 9.5% by mass of oseltamivir phosphate is added for hydration. After sufficient hydration, the mixture is heated in a water bath at 50° C. and subjected to reduced pressure rotary evaporation, high pressure homogenization, and freeze-dried to obtain a composition containing oseltamivir phosphate.

[0048] Examples 4-6 Oseltamivir phosphate tablets (1000 tablets)

[0049] The compositions containing oseltamivir phosphate of Examples 1 to 3 (containing 15 g of oseltamivir) were respectively taken, and 100 g of starch, 50 g of microcrystalline cellulose, and 20 g of sodium starch glycolate were crushed and mixed, and then 5 g of magnesium stearate was added. After mixing, the mixture was directly tableted to obtain the oseltamivir phosphate tablets of Examples 4 to 6.

[0050] Examples 7-9 Oseltamivir phosphate capsules (1000 tablets)

[0051] The compositions containing oseltamivir phosphate of Examples 1 to 3 (containing 15 g of oseltamivir) were respectively taken, and 150 g of dextrin, 20 g of mannitol, 15 g of low-substituted hydroxypropyl methylcellulose, and 30 g of sucrose were crushed and sieved and mixed, and an appropriate amount of starch slurry was added to prepare a soft material, granulated, dried, granulated, and filled into capsules to obtain the oseltamivir phosphate capsules of Examples 4 to 6.

[0052] Examples 10-12 Oseltamivir phosphate oral solution

[0053] Take the oseltamivir phosphate compositions of Examples 1 to 3 (containing 15 g of oseltamivir), add 50 g of syrup, 5 g of ethylparaben, 100 g of propylene glycol, and add purified water to the total amount to obtain the oseltamivir phosphate oral solutions of Examples 10 to 12.

[0054] Comparative Example 1 Composition of Oseltamivir Phosphate

[0055] prescription:

[0056] Dosage(g) Oseltamivir phosphate 20 Soybean lecithin 25 Egg yolk lecithin 10 cholesterol 12

[0057] Preparation method:

[0058] (1) Dissolve soybean lecithin, egg yolk lecithin and cholesterol in a methanol-ethanol mixed solvent with a volume ratio of 4:1, place in an eggplant-shaped bottle, heat in a water bath at 35°C and evaporate under reduced pressure to remove the solvent;

[0059] (2) After the film is formed, a glucose-phosphate buffer solution mixture (pH=8.0) containing 6.5% by mass of oseltamivir phosphate is added for hydration. After sufficient hydration, the mixture is heated in a water bath at 45° C. and subjected to reduced pressure rotary evaporation, high pressure homogenization, and freeze-dried to obtain a composition containing oseltamivir phosphate.

[0060] Comparative Example 2 Composition of Oseltamivir Phosphate

[0061] prescription:

[0062]

[0063] Preparation method:

[0064] (1) Menthone, soybean lecithin, egg yolk lecithin, and cholesterol were dissolved in a methanol-ethanol mixed solvent with a volume ratio of 4:1, placed in an eggplant-shaped bottle, heated in a water bath at 35°C and evaporated under reduced pressure to remove the solvent;

[0065] (2) After the film is formed, a glucose-phosphate buffer solution mixture (pH=8.0) containing 6.5% by mass of oseltamivir phosphate is added for hydration. After sufficient hydration, the mixture is heated in a water bath at 45° C. and subjected to reduced pressure rotary evaporation, high pressure homogenization, and freeze-dried to obtain a composition containing oseltamivir phosphate.

[0066] Comparative Example 3 Composition of Oseltamivir Phosphate

[0067] prescription:

[0068] Dosage(g) Oseltamivir phosphate 20 Pulegone 3 Soybean lecithin 25 Egg yolk lecithin 10 cholesterol 12

[0069] Preparation method:

[0070] (1) Pulegone, soybean lecithin, egg yolk lecithin, and cholesterol were dissolved in a methanol-water mixed solvent with a volume ratio of 4:1, placed in an eggplant-shaped bottle, heated in a water bath at 35°C and evaporated under reduced pressure to remove the solvent;

[0071] (2) After the film is formed, a phosphate buffer solution (pH=8.0) containing oseltamivir phosphate is added for hydration. After sufficient hydration, the film is heated in a water bath at 45° C. and then subjected to reduced pressure rotary evaporation, high pressure homogenization, and freeze drying to obtain a composition containing oseltamivir phosphate.

[0072] Comparative Example 4 Composition of Oseltamivir Phosphate

[0073] prescription:

[0074]

[0075]

[0076] Preparation method:

[0077] (1) Dissolve pulegone, soybean lecithin, egg yolk lecithin, and cholesterol in propylene glycol, place in an eggplant-shaped bottle, heat in a water bath at 35° C., and evaporate under reduced pressure to remove the solvent;

[0078] (2) After the film is formed, a glucose-phosphate buffer solution mixture (pH=8.0) containing 6.5% by mass of oseltamivir phosphate is added for hydration. After sufficient hydration, the mixture is heated in a water bath at 45° C. and subjected to reduced pressure rotary evaporation, high pressure homogenization, and freeze-dried to obtain a composition containing oseltamivir phosphate.

[0079] Comparative Examples 5-6 Oseltamivir Phosphate Capsules (1000 tablets)

[0080] The compositions containing oseltamivir phosphate of Comparative Example 1 and Comparative Example 3 (containing 15 g of oseltamivir) were respectively taken, and 150 g of dextrin, 20 g of mannitol, 15 g of low-substituted hydroxypropyl methylcellulose, and 30 g of sucrose were crushed and sieved and mixed, and an appropriate amount of starch slurry was added to prepare a soft material, granulated, dried, granulated, and filled into capsules to obtain the oseltamivir phosphate capsules of Examples 4 to 6.

[0081] Quality evaluation of compositions containing oseltamivir phosphate

[0082] The quality of the compositions containing oseltamivir phosphate of Examples 1 to 3 and the compositions containing oseltamivir phosphate of Comparative Examples 1 to 4 was evaluated, including drug loading, encapsulation efficiency, and leakage rate (the drug refers to oseltamivir phosphate), wherein the drug loading was measured by sampling on the first day and at the end of the sixth month of the accelerated test, the encapsulation efficiency was measured at the end of the sixth month of the accelerated test, and the leakage rate was measured by sampling at the end of the sixth month after the accelerated test was stored for 6 months.

[0083] Accelerated test conditions: Store at 40℃±2℃ and relative humidity 75%±5%.

[0084] Figure 2 The figure is a comparison of the drug loading of the compositions containing oseltamivir phosphate in Examples 1 to 3 and the compositions containing oseltamivir phosphate in Comparative Examples 1 to 4. Figure 3 The encapsulation efficiency and leakage rate of the compositions containing oseltamivir phosphate in Examples 1 to 3 and the compositions containing oseltamivir phosphate in Comparative Examples 1 to 4 are shown. The experimental results show that the compositions containing oseltamivir phosphate in Examples 1 to 3 of the present invention have high quality, high drug loading, high encapsulation efficiency, and low leakage rate, and in particular, the addition of glucose and the optimization of the solvent are beneficial to improving the quality of the liposome.

[0085] Single factor experiment to explore the effect of glucose concentration on the leakage rate of oseltamivir phosphate combination

[0086] The same composition containing oseltamivir phosphate as in Example 1, the variable factor being a 6.5% by mass mixture of glucose-phosphate buffer solution, and the concentrations thereof were 0.5%, 1.5%, 2.5%, 3.5%, 4.5%, 5.5%, 6.5%, 7.5%, 8.5%, 9.5%, 10.5%, 11.5%, 12.5%, and 15%, respectively. The prepared composition containing oseltamivir phosphate was subjected to accelerated test, and samples were taken at the end of the 6th month to determine and calculate the leakage rate.

[0087] Figure 4 The effect of glucose concentration on the leakage rate of the composition of oseltamivir phosphate is shown in Figure 1. The results show that only in a glucose solution with an appropriate mass fraction can the structural integrity of the liposome and the stability of the drug be maintained, and the leakage rate of the composition of oseltamivir phosphate be reduced.

[0088] Single factor experiment to explore the effect of glucose concentration on the leakage rate of oseltamivir phosphate combination

[0089] The same composition containing oseltamivir phosphate as in Example 1 was prepared, except that the pH of the phosphate buffer solution was 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, and 8.5, respectively. The prepared composition containing oseltamivir phosphate was subjected to an accelerated test, and samples were taken at the end of the 6th month to determine and calculate the leakage rate.

[0090] Figure 5 The results show that pH has an effect on the leakage rate of the composition of oseltamivir phosphate. The composition of oseltamivir phosphate may lose stability under acidic or neutral pH conditions, especially under acidic conditions, which may cause the dissociation or denaturation of the liposome structure, thereby causing leakage of the active ingredient. Figure 2 to Figure 5The experimental results show that the pH environment and the mass fraction of glucose complement each other and jointly affect the stability of the composition of oseltamivir phosphate.

[0091] Determination of related substances in oseltamivir phosphate

[0092] The relevant substances were determined according to the HPLC method (General Rule 0512) of the 2020 edition of the Chinese Pharmacopoeia. Solvent: 0.003 mol / L phosphoric acid solution-methanol-acetonitrile (620:245:135). Test solution: Take about 50 mg of the test sample, accurately weigh it, place it in a 50 ml volumetric flask, add solvent to dissolve and dilute to the scale, and shake well. Control solution: Accurately measure an appropriate amount of the test solution, and quantitatively dilute it with the mobile phase to make a solution containing about 1 μg per 1 ml. Reference solution: Take an appropriate amount of impurity I reference substance, impurity II reference substance, and impurity III reference substance, accurately weigh them respectively, dissolve them with solvent, and quantitatively dilute them to make a solution containing about 1.0 μg of impurity I, 1.0 μg of impurity II, and 2.0 μg of impurity III per 1 ml. Chromatographic conditions: Octylsilane bonded silica gel is used as filler; 0.05mol / L potassium dihydrogen phosphate solution (adjust pH to 5.6 with 1mol / L potassium hydroxide solution)-methanol-acetonitrile (700:245:135) is used as mobile phase; flow rate is 1.0ml per minute; detection wavelength is 207nm; column temperature is 50℃; injection volume is 15μl. System suitability requirements: In the chromatogram of the reference solution, the separation between the peaks of impurities Ⅰ, impurity Ⅱ and impurity Ⅲ should meet the requirements. Determination method: Accurately measure the test solution, reference solution and reference solution, inject them into the liquid chromatograph respectively, and record the chromatogram until twice the retention time of the main component peak. Limit: If there are impurity peaks in the chromatogram of the test solution, impurities Ⅰ, impurity Ⅱ and impurity Ⅲ calculated by peak area according to the external standard method shall not exceed 0.1%, 0.1% and 0.2% respectively, and the total amount of impurities shall not exceed 0.5%.

[0093] The oseltamivir phosphate preparations of Examples 4 to 9 and the oseltamivir phosphate capsules of Comparative Example 6 were subjected to an accelerated test at a temperature of 40°C ± 2°C, a relative humidity of 75% ± 5%, and an illumination of 4500 lx ± 500 lx. Samples were taken on the first day, at the end of the second month, at the end of the third month, and at the end of the sixth month, and the contents of related substances were determined.

[0094] Fig. 9 The graphs showing changes in the contents of related substances in the oseltamivir phosphate preparations of Examples 4 to 9 and the oseltamivir phosphate capsules of Comparative Example 6 show that the oseltamivir phosphate preparations of the present invention have high stability, low contents of related substances, and high safety of medication. In addition, it also reveals that glucose can improve the stability of the oseltamivir phosphate liposomes, thereby improving the stability of the preparation and preventing the generation of related substances.

[0095] Pharmacodynamics experiments

[0096] 1. Experimental Animals

[0097] SD rats SPF grade, 180-220 g, experimental animal license number: SYXK(Lu)20180008, were adaptively fed under standard conditions for 1 week before the experiment.

[0098] 2. Modeling method and grouping

[0099] Ether inhalation anesthesia was used, and 0.1 ml of AIV liquid with a median lethal dose (LD50) was alternately dripped into the left and right nostrils of the rats to establish an AIV infection rat model (A / FM1 / 47 (H1N1) is an AIV mouse lung-adapted strain, frozen in the virus room of Shandong University of Traditional Chinese Medicine). The normal control group rats were anesthetized by ether inhalation and inoculated with an equal amount of virus-free physiological saline. The rats with successful modeling were divided into a model group, Example 4 group, Example 5 group, Example 6 group, Comparative Example 5 group, and a commercially available group, with 10 rats in each group.

[0100] 3. Drug administration

[0101] Example 4 group: Example 4 tablets, calculated as oseltamivir phosphate, with a dosage of 5.4 kg / kg, were administered by oral gavage twice a day for 5 consecutive days.

[0102] Example 5 group: Example 5 tablets, calculated as oseltamivir phosphate, with a dosage of 5.4 kg / kg, were administered by oral gavage twice a day for 5 consecutive days.

[0103] Example 6 group: Example 6 tablets, calculated as oseltamivir phosphate, with a dosage of 5.4 kg / kg, were administered by oral gavage twice a day for 5 consecutive days.

[0104] Comparative Example 5 group: Comparative Example 1 tablets, calculated as oseltamivir phosphate, with a dosage of 5.4 kg / kg, were administered orally twice a day for 5 consecutive days.

[0105] Commercially available group: Oseltamivir phosphate capsules (H20140344), based on oseltamivir phosphate, the dose was 5.4 kg / kg, administered by gavage twice a day for 5 consecutive days.

[0106] Model group: The same volume of normal saline was administered orally twice a day for 5 consecutive days.

[0107] Blank group: The same amount of normal saline was administered intragastrically twice a day for 5 consecutive days.

[0108] 4. Statistical analysis

[0109] SPSS 22.0 software was used to perform statistical analysis on the data. The measurement data were expressed as (x±s). One-way analysis of variance was used for comparison among multiple groups, and independent sample T test was used for analysis between two groups. P<0.05 was considered statistically significant.

[0110] 5. Rat body temperature statistics

[0111] The body temperature of rats in each group was measured 2h, 6h, 12h, 24h, 48h and 60h after the first administration, and statistics were performed.

[0112] The results are as follows Fig.10 As shown, compared with the blank group, the body temperature of the model group was significantly increased, indicating that the model was successfully established. The body temperature of the rats in each medication group was significantly lower than that in the model group. The body temperature of the rats in Examples 4 to 6 groups decreased significantly, indicating that the oseltamivir phosphate capsules in Examples 4 to 6 had a significant inhibitory effect on the body temperature of rats infected with influenza A IV, and the effect was better than that of the commercially available preparation group and the comparative example 5 group ( Fig.10 Comparative Example 1) in the present invention confirms that the combination of oseltamivir phosphate and pulegone has a synergistic effect.

[0113] 6. TNF-α and IL-1β levels in rat serum

[0114] The cytokines, TNF-α and IL-1β in serum were detected by ELISA.

[0115] Table 1 TNF-α and IL-1β levels in rat serum

[0116]

[0117] As shown in Table 1, compared with the commercially available group, the levels of TNF-α and IL-1β in the serum of rats in groups 4 to 6 of Examples were significantly different, *P<0.01; compared with the comparative example 5 group, the levels of TNF-α and IL-1β in the serum of rats in groups 4 to 6 of Examples were significantly different, *P<0.01. This shows that the oseltamivir phosphate preparation of the present invention has a better therapeutic effect on rats infected with type A IV. It should be noted that the reason for preparing the oseltamivir phosphate composition into a preparation for pharmacodynamic experiments in the present invention is to compare the therapeutic effects of the oseltamivir phosphate composition of the present invention with those of the commercially available preparations. The animal experiments of the oseltamivir phosphate composition of comparative example 2 have been carried out, and its effects are similar to the various indicators of the rats in group 5 of comparative example.

Claims

1. A composition containing oseltamivir phosphate, characterized in that: The preparation method of the composition containing oseltamivir phosphate is as follows: 1-5 parts by weight of pulegone, 15-50 parts by weight of soybean lecithin or / and egg yolk lecithin, and 5-20 parts by weight of cholesterol are dissolved in a methanol-ethanol mixed solvent with a volume ratio of 3-5:1, placed in an eggplant-shaped bottle, heated in a water bath and subjected to reduced pressure rotary evaporation to remove the organic solvent, and a glucose-buffer solution mixture with a pH of 7.5-8.5 and a mass fraction of 4.5%-9.5% dissolved in 10-40 parts by weight of oseltamivir phosphate is added for hydration, and after sufficient hydration, heated in a water bath and subjected to reduced pressure rotary evaporation, high pressure homogenization, and freeze drying are continued to obtain the composition containing oseltamivir phosphate.

2. The composition containing oseltamivir phosphate according to claim 1, characterized in that: The water bath temperature is 30°C to 50°C.

3. A preparation, characterized in that The preparation consists of the composition containing oseltamivir phosphate according to claim 1 and pharmaceutically acceptable excipients.

4. The preparation according to claim 3, characterized in that The preparation is one or more of tablets, granules, capsules, pills, and oral liquids.

5. The preparation according to claim 3, characterized in that The pharmaceutically acceptable excipient is selected from at least one of a filler, a lubricant, a disintegrant, a binder, a humectant, a solvent, a flavoring agent, a preservative, a stabilizer, a thickener, and a surfactant.

6. Use of the composition containing oseltamivir phosphate according to claim 1 in the preparation of anti-influenza A virus drugs.

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