Arbidol hydrochloride sustained release tablet as well as preparation method and application thereof

Abstract

The invention provides arbidol hydrochloride sustained release tablets as well as a preparation method and application thereof, and belongs to the technical field of medicines. The preparation method comprises the following steps: performing water extraction and alcohol precipitation on liquorice, ginger and flos puerariae to obtain hydrophilic active polysaccharide, performing ionic liquid separation to obtain an active component, mixing arbidol hydrochloride, biotoxin and the active component to prepare liposome, uniformly mixing the liposome with lactose, cross-linked povidone and the hydrophilic active polysaccharide, and preparing quick-release particles by using a povidone K30 solution and magnesium stearate; mixing arbidol hydrochloride and microcrystalline cellulose, and preparing sustained-release particles by using ethanol and magnesium stearate; the arbidol hydrochloride sustained-release tablet has the double properties of sustained release and quick release, the sustained-release effect reduces the administration frequency, and the quick-release effect can enable the blood concentration to quickly rise, so that the drug effect is quickly exerted to generate a treatment effect, the defect that a common sustained-release preparation takes effect slowly is overcome, and the arbidol hydrochloride sustained-release tablet is suitable for clinical application. The preparation method is simple, convenient and suitable for industrial production.

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to an arbidol hydrochloride sustained-release tablet and a preparation method and application thereof. Background Art

[0002] Arbidol is a highly effective drug for preventing and treating influenza A and B and other acute respiratory viral infections. Its mechanism of action is different from that of commonly used antiviral drugs in clinical practice, such as ribavirin, amantadine and rimantadine. It fights against influenza viruses by inducing interferon and enhancing immune function. In addition, it has antagonistic effects on influenza A and B viruses, has a wider antiviral spectrum than amantadine, and can activate macrophages, effectively treating influenza and other acute respiratory viral infections. Recent studies have shown that the drug also has a good inhibitory effect on atypical viruses at a lower safety concentration. Common dosage forms developed in China include tablets, capsules, dispersible tablets, granules, etc., which are taken 3-4 times a day, and each tablet contains 100mg or 200mg.

[0003] At present, the dosage forms of Abidor on the market include capsules, tablets, and oral suspensions. Due to the rapid release of the active ingredients in the drug, the blood drug concentration fluctuates greatly, and the effectiveness and safety of the drug cannot be guaranteed. In addition, patients need to take the drug many times, which makes it inconvenient for patients with chronic diseases who take the drug for a long time. At the same time, Abidor has poor water solubility, and sustained-release preparations often have the phenomenon of incomplete final drug release.

[0004] Chinese invention patent application CN1589790A discloses a sustained-release tablet involving the antiviral drug Arbidol, which uses a hydrophilic gel skeleton material or a wax skeleton material to prepare a sustained-release tablet containing Arbidol. However, a simple sustained-release tablet releases the drug slowly and cannot quickly release the active ingredient of the drug, resulting in a low maximum blood drug concentration. Summary of the invention

[0005] The purpose of the present invention is to provide an arbidol hydrochloride sustained-release tablet and a preparation method and application thereof, which have both sustained-release and rapid-release properties. The sustained-release effect can reduce the number of dosing times, and the rapid-release effect can rapidly increase the blood drug concentration, thereby rapidly exerting the drug efficacy to produce a therapeutic effect, overcoming the shortcoming of slow onset of ordinary sustained-release preparations. The preparation method of the present invention is simple and convenient, and is suitable for industrial production.

[0006] The technical solution of the present invention is achieved in this way:

[0007] The invention provides a method for preparing an arbidol hydrochloride sustained-release tablet. The method comprises the following steps: extracting liquorice, ginger and kudzu flower with water and performing alcohol precipitation to obtain hydrophilic active polysaccharides, separating active components by ionic liquid, mixing arbidol hydrochloride, biological toxins and active components to prepare liposomes, uniformly mixing the liposomes with lactose, cross-linked polyvinylpyrrolidone and the hydrophilic active polysaccharides, and preparing quick-release granules with polyvinylpyrrolidone K30 solution and magnesium stearate; mixing arbidol hydrochloride and microcrystalline cellulose, and preparing slow-release granules with ethanol and magnesium stearate; and tableting the quick-release granules and the slow-release granules in layers to prepare the arbidol hydrochloride sustained-release tablets.

[0008] As a further improvement of the present invention, the following steps are included:

[0009] S1. Extraction of hydrophilic active polysaccharides: Licorice, ginger and kudzu flower are washed, dried and crushed respectively, added into water, heated to boiling for extraction, filtered, the solid residue is retained, ethanol is added into the liquid for precipitation, filtered, the filtrate is retained, the solid is washed and dried to obtain the hydrophilic active polysaccharide;

[0010] S2. Extraction of active components: The solid residue in step S1 and the filtrate in step S1 are mixed and added to the ionic liquid, carbon dioxide is introduced, stirred for extraction, filtered, the filtrate is heated to allow carbon dioxide to escape, separated, the organic layer is collected, the active component is obtained, and the ionic liquid layer is washed and reused;

[0011] S3. Preparation of liposomes: Abidol hydrochloride, biotoxins, and active ingredients were mixed uniformly, lecithin and glyceryl monostearate were added, and the mixture was stirred and dispersed uniformly to obtain a mixture; an aqueous solution containing polysorbate-80 and polyethylene glycol 400 was prepared, the mixture was added dropwise with stirring, ultrasonically dispersed uniformly, and freeze-dried to obtain liposomes;

[0012] S4. Preparation of immediate-release granules: liposomes, lactose, cross-linked polyvinylpyrrolidone, and hydrophilic active polysaccharide are uniformly mixed, a soft material is prepared with a polyvinylpyrrolidone K30 solution, granulated, dried, granulated, and magnesium stearate is added and mixed to prepare immediate-release granules;

[0013] S5. Preparation of sustained-release granules: Abidol hydrochloride and microcrystalline cellulose were mixed and sieved, ethanol was added to prepare a soft material, granulated, dried, granulated, magnesium stearate was added and mixed, and sustained-release granules were prepared;

[0014] S6. Preparation of Abidor Hydrochloride Sustained-Release Tablets: The immediate-release granules and the sustained-release granules are layered and tableted in proportion to prepare Abidor Hydrochloride Sustained-Release Tablets.

[0015] As a further improvement of the present invention, the mass ratio of licorice, ginger and kudzu flower in step S1 is 3-5:5-7:10-15, and the time for heating to boiling extraction is 3-5h.

[0016] As a further improvement of the present invention, the ionic liquid in step S2 is selected from at least one of 1-butyl-3-methylimidazolium hexafluorophosphate, 1-butyl-3-methylimidazolium bistrifluoromethanesulfonyl imide salt, 1-butyl-3-methylimidazolium hexafluoroantimonate, 1-hexyl-3-methylimidazolium tetrafluoroborate, 1-hexyl-3-methylimidazolium hexafluorophosphate, and 1-hexyl-3-methylimidazolium hexafluoroantimonate, the carbon dioxide ventilation rate is 10-20 mL / min, the stirring extraction time is 20-30 min, and the filtrate heating temperature is 50-60 ° C.

[0017] As a further improvement of the present invention, the biotoxin in step S3 is selected from at least one of bee venom peptide, phospholipase A2, and bee venom peptide, the mass ratio of arbidol hydrochloride, biotoxin, active ingredient, lecithin and glyceryl monostearate is 15-20:3-5:4-6:25-35:100-150, and the concentration of polysorbate-80 in the aqueous solution containing polysorbate-80 and polyethylene glycol 400 is 0.5-1wt%, and the concentration of polyethylene glycol 400 is 1-2wt%.

[0018] As a further improvement of the present invention, the mass ratio of the liposomes, lactose, cross-linked polyvinylpyrrolidone, hydrophilic active polysaccharide, polyvinylpyrrolidone K30 solution, and magnesium stearate in step S4 is 10-15:4-7:15-20:3-5:30-50:0.5-1, and the concentration of the polyvinylpyrrolidone K30 solution is 5-10wt%.

[0019] As a further improvement of the present invention, the mass ratio of arbidol hydrochloride, microcrystalline cellulose, ethanol and magnesium stearate in step S5 is 8-10:4-7:15-20:0.5-1, and the mesh number of the sieve is 80-100 mesh.

[0020] As a further improvement of the present invention, the mass ratio of the rapid-release granules to the sustained-release granules in step S6 is 3-5:2-4.

[0021] The present invention further protects an arbidol hydrochloride sustained-release tablet prepared by the above-mentioned preparation method.

[0022] The present invention further protects the use of the above-mentioned arbidol hydrochloride sustained-release tablets in the preparation of medicines for treating pneumonia and influenza.

[0023] The present invention has the following beneficial effects:

[0024] Pueraria irisin is the main bioactive component of Pueraria iris isoflavones. It is a polyphenolic hydroxyl compound with multiple functions such as anti-oxidative damage, lowering blood lipids, protecting the liver, anti-virus, preventing and treating cardiovascular diseases, etc. It has important health and medicinal value. However, irisin has poor water solubility and low bioavailability. By preparing it into liposomes, its biocompatibility and bioavailability are greatly improved. Pueraria flowers, licorice and ginger are rich in polysaccharides, and other active components include liquiritigenin, liquiritigenin, isoliquiritigenin, etc. Ginger is rich in gingerol, shogaol, shogaol, gingerol, zingerone and gingerdione, etc. The hydrophilic active polysaccharides obtained by water extraction and alcohol precipitation have good synergistic antiviral activity on the one hand, and can directly interact with the virus surface through the negative charge it carries, thereby inhibiting the virus's infection ability, or directly killing the virus and making it lose its infectivity. On the other hand, it can be synergistically prepared with cross-linked polyvinylpyrrolidone and polyvinylpyrrol K30 for rapid-release granules, which are non-toxic, environmentally friendly, renewable, biodegradable and biocompatible. The release rate and release time of the drug can be controlled through its physical and chemical properties to achieve a controlled release effect.

[0025] The present invention uses carbon dioxide to increase the polarity of the ionic liquid. The anions of the ionic liquid can adsorb and activate CO2, and activate other molecules through hydrogen bonding and induction effects, thereby improving the extraction rate of the ionic liquid for the active components in the traditional Chinese medicine components. After heating, the CO2 molecules are dissipated, thereby reducing the polarity of the ionic liquid, so that the active components and the ionic liquid are stratified, thereby separating the active components well. The active components include liquiritigenin, liquiritigenin, isoliquiritigenin, etc. Ginger is rich in gingerol, shogaol, shogaol, gingerol, zingerone and gingerdione, etc., which can target the metal ion binding sites of metal ion binding proteins, and can effectively interfere with the life cycle of the virus, thereby inhibiting its infection and spread.

[0026] Antimicrobial peptides in animal toxins can directly kill viruses, or play an antiviral role by regulating the immune system, inhibiting viral replication, etc. The bee venom selected in the present invention can affect intracellular calcium ion metabolism, and destroy membrane polarization by combining with calcium-dependent potassium ion channels in cells, thereby playing a good auxiliary antiviral role.

[0027] Arbidol can be combined with cell membrane lipid bilayer and mixed with lipid bilayer, in the micromolar range, can be connected with the phospholipid pole head of boundary membrane surface, by the interaction with phospholipid, the plasma membrane fluidity will be disturbed, and the lipid bilayer is more difficult to merge. Its indole structure can form supramolecular arrangement by carrying out aromatic stacking reaction with the selective amino acid residues of protein (phenylalanine, tyrosine, tryptophan). Therefore, Arbidol fills the gap between lipid molecules and adopts consistent orientation by inserting in the lipid of viral membrane and cell membrane, so that Arbidol is highly concentrated in the local area, and then interacts with the aromatic residues in viral protein. However, the water solubility, salt solubility and alkali solubility of Arbidol are poor, which greatly limits its application. In the liposomes prepared by the present invention, Abidol prevents the conformational rearrangement of the fusion protein locally through dual binding ability, effectively inhibits the fusion of the virus and the host cell membrane and blocks the invasion of the virus. At the same time, the biological toxin is targeted and brought into the vicinity of the virus, thereby obtaining a local high concentration of the biological toxin, exerting the effect of the biological toxin in destroying membrane polarization, thereby playing a good synergistic antiviral effect.

[0028] The surface membrane of influenza virus contains two glycoproteins, hemagglutinin and neuraminidase, which play an important role in the invasion process of influenza virus. Hemagglutinin is responsible for binding the virus to the sialic acid receptor on the cell surface, thereby starting the infection process. After the virus completes replication and assembly, neuraminidase removes the sialic acid connecting the virus and the cell surface glycoprotein, promoting the release and spread of the virus. These two glycoproteins are like two keys, hemagglutinin opens the door of the cell, and neuraminidase releases the chain that the virus is bound to the cell. Due to the presence of hydrophobic pockets in the active site of neuraminidase, the present invention prepares liposomes, which improves the water solubility of the drug on the one hand, greatly improves the biocompatibility of the drug, and on the other hand, it can also improve its lipophilicity to the hydrophobic pocket of neuraminidase, so that the liposome can be targeted and delivered to the vicinity of the virus, and the local drug concentration is increased, thereby greatly improving the efficacy of the drug.

[0029] The invention provides an arbidol multilayer sustained-release tablet, which has both sustained-release and rapid-release properties. The sustained-release effect can reduce the number of drug administrations, and the rapid-release effect can rapidly increase the blood drug concentration, thereby rapidly exerting the drug efficacy to produce a therapeutic effect, thereby overcoming the shortcoming of slow onset of action of common sustained-release preparations. The preparation method of the invention is simple and convenient, and is suitable for industrial production. DETAILED DESCRIPTION

[0030] The technical solutions in the embodiments of the present invention are described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0031] Example 1

[0032] This embodiment provides an arbidol hydrochloride sustained-release tablet, comprising the following steps:

[0033] S1. Extraction of hydrophilic active polysaccharides: 3g of licorice, 5g of ginger, and 10g of kudzu flower were washed, dried, and crushed, added to 200mL of water, heated to boiling for extraction for 3h, filtered, and the solid residue was retained. Ethanol was added to the liquid until the ethanol content of the system was 80wt%, precipitated for 1h, filtered, and the filtrate was retained. The solid was washed and dried to obtain a hydrophilic active polysaccharide;

[0034] S2. Extraction of active components: The solid residue in step S1 and the filtrate in step S1 were mixed and added to 100 mL of ionic liquid 1-butyl-3-methylimidazolium hexafluorophosphate, carbon dioxide was introduced at a ventilation rate of 10 mL / min, and the mixture was stirred for 20 min, filtered, and the filtrate was heated to 50 ° C, stirred to allow carbon dioxide to escape, separated, the organic layer was collected, and dried to obtain the active component. The ionic liquid layer was washed and reused;

[0035] S3. Preparation of liposomes: 1.5 g of arbidol hydrochloride, 0.3 g of bee venom peptide, and 0.4 g of the active ingredient were mixed for 10 minutes, 2.5 g of lecithin and 10 g of glyceryl monostearate were added, and the mixture was added to 150 mL of ethanol, and stirred and dispersed for 10 minutes to obtain a mixture; an aqueous solution containing polysorbate-80 and polyethylene glycol 400 (the concentration of polysorbate-80 was 0.5 wt %, and the concentration of polyethylene glycol 400 was 1 wt %) was prepared, the mixture was added dropwise with stirring, ultrasonically dispersed at 1000 W for 15 minutes, and freeze-dried to obtain liposomes;

[0036] S4. Preparation of immediate-release granules: 1 g of liposomes, 0.4 g of lactose, 1.5 g of cross-linked polyvinylpyrrolidone, and 0.3 g of hydrophilic active polysaccharide were mixed for 15 min, 3 g of 5 wt% polyvinylpyrrolidone K30 solution was used to prepare a soft material, granulated, dried, granulated, 0.05 g of magnesium stearate was added and mixed for 10 min to prepare immediate-release granules;

[0037] S5. Preparation of sustained-release granules: 0.8 g of arbidol hydrochloride and 0.4 g of microcrystalline cellulose were mixed and passed through an 80-mesh sieve, 1.5 g of ethanol was added to prepare a soft material, granulated, dried, granulated, 0.05 g of magnesium stearate was added and mixed for 10 min to obtain sustained-release granules;

[0038] S6. Preparation of Abidor Hydrochloride Sustained-Release Tablets: 3 g of immediate-release granules and 2 g of sustained-release granules were layered and tableted in proportion to prepare Abidor Hydrochloride Sustained-Release Tablets.

[0039] Example 2

[0040] This embodiment provides an arbidol hydrochloride sustained-release tablet, comprising the following steps:

[0041] S1. Extraction of hydrophilic active polysaccharides: 5g of licorice, 7g of ginger, and 15g of kudzu flower were washed, dried, and crushed, added to 200mL of water, heated to boiling and extracted for 5h, filtered, and the solid residue was retained. Ethanol was added to the liquid until the ethanol content of the system was 80wt%, precipitated for 1h, filtered, and the filtrate was retained. The solid was washed and dried to obtain a hydrophilic active polysaccharide;

[0042] S2. Extraction of active components: The solid residue in step S1 and the filtrate in step S1 were mixed and added to 100 mL of ionic liquid 1-butyl-3-methylimidazolium hexafluoroantimonate, carbon dioxide was introduced at a ventilation rate of 20 mL / min, and the mixture was stirred for 30 min, filtered, and the filtrate was heated to 60 ° C, stirred to allow carbon dioxide to escape, separated, the organic layer was collected, and dried to obtain the active component. The ionic liquid layer was washed and reused;

[0043] S3. Preparation of liposomes: 2 g of arbidol hydrochloride, 0.5 g of bee venom peptide, and 0.6 g of the active ingredient were mixed for 10 min, 3.5 g of lecithin and 15 g of glyceryl monostearate were added, and the mixture was added to 150 mL of ethanol, and stirred and dispersed for 10 min to obtain a mixture; an aqueous solution containing polysorbate-80 and polyethylene glycol 400 (the concentration of polysorbate-80 was 1 wt%, and the concentration of polyethylene glycol 400 was 2 wt%) was prepared, the mixture was added dropwise with stirring, ultrasonically dispersed at 1000 W for 15 min, and freeze-dried to obtain liposomes;

[0044] S4. Preparation of immediate-release granules: 1.5 g liposomes, 0.7 g lactose, 2 g cross-linked polyvinylpyrrolidone, and 0.5 g hydrophilic active polysaccharide were mixed for 15 min, 5 g 10 wt% polyvinylpyrrolidone K30 solution was used to prepare a soft material, granulated, dried, granulated, 0.1 g magnesium stearate was added and mixed for 10 min to prepare immediate-release granules;

[0045] S5. Preparation of sustained-release granules: 1 g of arbidol hydrochloride and 0.7 g of microcrystalline cellulose were mixed and passed through a 100-mesh sieve, 2 g of ethanol was added to prepare a soft material, granulated, dried, granulated, 0.1 g of magnesium stearate was added and mixed for 10 min to prepare sustained-release granules;

[0046] S6. Preparation of Abidor Hydrochloride Sustained-Release Tablets: 5 g of immediate-release granules and 4 g of sustained-release granules were layered and tableted in proportion to prepare Abidor Hydrochloride Sustained-Release Tablets.

[0047] Example 3

[0048] This embodiment provides an arbidol hydrochloride sustained-release tablet, comprising the following steps:

[0049] S1. Extraction of hydrophilic active polysaccharides: 4 g of licorice, 6 g of ginger, and 12 g of kudzu flower were washed, dried, and crushed, added to 200 mL of water, heated to boiling and extracted for 4 h, filtered, and the solid residue was retained. Ethanol was added to the liquid until the ethanol content of the system was 80 wt%, precipitated for 1 h, filtered, the filtrate was retained, the solid was washed, and dried to obtain a hydrophilic active polysaccharide;

[0050] S2. Extraction of active components: The solid residue in step S1 and the filtrate in step S1 were mixed and added to 100 mL of ionic liquid 1-hexyl-3-methylimidazolium hexafluorophosphate, carbon dioxide was introduced at a ventilation rate of 15 mL / min, and the mixture was stirred for 25 min, filtered, and the filtrate was heated to 55 ° C, stirred to allow carbon dioxide to escape, separated, the organic layer was collected, and dried to obtain the active component. The ionic liquid layer was washed and reused;

[0051] S3. Preparation of liposomes: 1.7 g of arbidol hydrochloride, 0.4 g of bee venom peptide, and 0.5 g of the active ingredient were mixed for 10 min, 3 g of lecithin and 12 g of glyceryl monostearate were added, and the mixture was added to 150 mL of ethanol, and stirred and dispersed for 10 min to obtain a mixture; an aqueous solution containing polysorbate-80 and polyethylene glycol 400 (the concentration of polysorbate-80 was 0.7 wt %, and the concentration of polyethylene glycol 400 was 1.5 wt %) was prepared, the mixture was added dropwise with stirring, ultrasonically dispersed at 1000 W for 15 min, and freeze-dried to obtain liposomes;

[0052] S4. Preparation of immediate-release granules: 1.2 g of liposomes, 0.55 g of lactose, 1.7 g of cross-linked polyvinylpyrrolidone, and 0.4 g of hydrophilic active polysaccharide were mixed for 15 min, 4 g of 7 wt % polyvinylpyrrolidone K30 solution was used to prepare a soft material, granulated, dried, granulated, 0.07 g of magnesium stearate was added and mixed for 10 min to prepare immediate-release granules;

[0053] S5. Preparation of sustained-release granules: 0.9 g of arbidol hydrochloride and 0.55 g of microcrystalline cellulose were mixed through a 100-mesh sieve, 1.7 g of ethanol was added to prepare a soft material, granulated, dried, granulated, 0.07 g of magnesium stearate was added and mixed for 10 min to obtain sustained-release granules;

[0054] S6. Preparation of Abidor Hydrochloride Sustained-Release Tablets: 4 g of immediate-release granules and 3 g of sustained-release granules were layered and tableted in proportion to prepare Abidor Hydrochloride Sustained-Release Tablets.

[0055] Comparative Example 1

[0056] Compared with Example 3, the difference is that no kudzu flower is added in step S1.

[0057] The details are as follows:

[0058] S1. Extraction of hydrophilic active polysaccharides: 4g licorice, 6g ginger and 12g kudzu flower were washed, dried and crushed respectively, added into 200mL water, heated to boiling and extracted for 4h, filtered, and the solid residue was retained. Ethanol was added to the liquid until the ethanol content of the system was 80wt%, precipitated for 1h, filtered, and the filtrate was retained. The solid was washed and dried to obtain a hydrophilic active polysaccharide.

[0059] Comparative Example 2

[0060] Compared with Example 3, the difference is that liquorice is not added in step S1.

[0061] The details are as follows:

[0062] S1. Extraction of hydrophilic active polysaccharides: Wash, dry and crush 6 g of ginger, add into 200 mL of water, heat to boiling and extract for 4 h, filter, retain the solid residue, add ethanol to the liquid until the ethanol content of the system is 80 wt%, precipitate for 1 h, filter, retain the filtrate, wash the solid, and dry to obtain the hydrophilic active polysaccharide.

[0063] Comparative Example 3

[0064] Compared with Example 3, the difference is that ginger is not added in step S1.

[0065] The details are as follows:

[0066] S1. Extraction of hydrophilic active polysaccharides: Wash, dry and crush 4g of licorice and 12g of kudzu flower respectively, add them into 200mL of water, heat to boiling and extract for 4h, filter, retain the solid residue, add ethanol to the liquid until the ethanol content of the system is 80wt%, precipitate for 1h, filter, retain the filtrate, wash the solid, and dry to obtain hydrophilic active polysaccharides.

[0067] Comparative Example 4

[0068] Compared with Example 3, the difference is that no melittin is added in step S3.

[0069] The details are as follows:

[0070] S3. Preparation of liposomes: Mix 1.7 g of arbidol hydrochloride and 0.5 g of active ingredient for 10 minutes, add 3 g of lecithin and 12 g of glyceryl monostearate, add into 150 mL of ethanol, stir and disperse for 10 minutes to obtain a mixture; prepare an aqueous solution containing polysorbate 80 and polyethylene glycol 400 (the concentration of polysorbate 80 is 0.7 wt%, the concentration of polyethylene glycol 400 is 1.5 wt%), add the mixture dropwise with stirring, ultrasonically disperse at 1000 W for 15 minutes, freeze-dry, and obtain liposomes.

[0071] Comparative Example 5

[0072] Compared with Example 3, the difference is that no active component is added in step S3.

[0073] The details are as follows:

[0074] S3. Preparation of liposomes: Mix 1.7 g of arbidol hydrochloride and 0.4 g of bee venom peptide for 10 minutes, add 3 g of lecithin and 12 g of glyceryl monostearate, add into 150 mL of ethanol, stir and disperse for 10 minutes to obtain a mixture; prepare an aqueous solution containing polysorbate-80 and polyethylene glycol 400 (the concentration of polysorbate-80 is 0.7wt%, and the concentration of polyethylene glycol 400 is 1.5wt%), stir and add dropwise the mixture, ultrasonically disperse at 1000W for 15 minutes, freeze-dry, and obtain liposomes.

[0075] Comparative Example 6

[0076] Compared with Example 3, the difference is that liposomes are not prepared in step S3.

[0077] The details are as follows:

[0078] S3. Preparation of a mixture: 1.7 g of arbidol hydrochloride, 0.4 g of melittin, and 0.5 g of an active ingredient were mixed for 10 min to prepare a mixture.

[0079] Comparative Example 7

[0080] Compared with Example 3, the difference is that no hydrophilic active polysaccharide is added in step S4.

[0081] The details are as follows:

[0082] S4. Preparation of rapid-release granules: 1.2 g liposomes, 0.55 g lactose, and 2.1 g cross-linked polyvinylpyrrolidone were mixed for 15 min, and a soft material was prepared with 4 g 7 wt % polyvinylpyrrolidone K30 solution. The mixture was granulated, dried, and granulated. 0.07 g magnesium stearate was added and mixed for 10 min to obtain rapid-release granules.

[0083] Test Example 1

[0084] According to the release determination method of the first method of Appendix XC of the Chinese Pharmacopoeia (Part II of the 2000 edition), 900 ml of 0.1 mol / L acetic acid was used as the release medium, the rotation speed was 100 r / min, and the release performance of the arbidol hydrochloride sustained-release tablets and ordinary arbidol hydrochloride tablets (6 tablets) prepared in Examples 1-3 and Comparative Example 7 was tested according to the law. 5 mL of samples were taken at 5, 10, 20 min and 0.5, 1, 2, 4, 6, 8, 10, and 12 h, and the same volume of isothermal release medium was added at the same time; 1 mL of the subsequent filtrate was taken, and the absorbance was measured after dilution with 0.mol / L acetic acid at different time points according to different dilution multiples, and the average release was calculated.

[0085] The results are shown in Table 1.

[0086] Table 1

[0087] As can be seen from the above table, the arbidol hydrochloride sustained-release tablets prepared in Examples 1-3 of the present invention have a rapid release effect within 1 hour, and can release the drug for up to 12 hours, while the ordinary arbidol hydrochloride tablets will be completely released within 1 hour. In Comparative Example 7, no hydrophilic active polysaccharide was added, and its rapid release effect within 1 hour decreased, which shows that the hydrophilic active polysaccharide has a good promoting effect on rapid release within 1 hour.

[0088] Test Example 2

[0089] 130 SPF-grade ICR mice (14 ± 2g) were selected, half male and half female, and randomly divided into 13 groups: normal control group, model control group, Tamiflu control group, Example 1-3 group, and Comparative Example 1-7 group. Each group contained 10 mice, half male and half female. The mice were lightly anesthetized with isoflurane. Except for the normal control group, the other groups were infected with 15 LD50 H1N1 / PR8 influenza virus liquids by nasal drops. The normal control group used normal saline, and each mouse was given 35 μL nasal drops. Starting on the day of infection, each group of mice was given oral administration of the corresponding Abidor Hydrochloride Sustained-release Tablets for drug treatment, with a dosage of 1g / kg, once a day, for 4 days, and the normal control group and the model control group were given an equal mass of 0.5wt% sodium carboxymethyl cellulose solution.

[0090] 1. Determination of lung index

[0091] On the 5th day of infection, the mice were weighed, dissected, and their lungs were removed and weighed to calculate the lung index of the mice and the inhibition rate of the drug.

[0092] Lung index (g / 100g) = lung wet weight (g) / body weight (100g)

[0093] The results are shown in Table 2.

[0094] Table 2 Group Lung index (g / 100g) Normal control group 0.70±0.24 Model control group 0.98±0.19* Tamiflu control group 0.79±0.21# Example 1 0.73±0.20# Example 2 0.74±0.17# Example 3 0.72±0.23# Comparative Example 1 0.78±0.29 Comparative Example 2 0.80±0.27 Comparative Example 3 0.81±0.25 Comparative Example 4 0.79±0.31 Comparative Example 5 0.84±0.26 Comparative Example 6 0.82±0.34 Comparative Example 7 0.80±0.30 Note: * compared with the normal control group, P < 0.05; # compared with the model control group, P < 0.05.

[0095] It can be seen from the above table that the Abidol Hydrochloride Sustained-Release Tablets prepared in Examples 1-3 of the present invention can significantly reduce the lung index of mice.

[0096] 2. Detecting lung viral load

[0097] Weigh 20-30 mg of mouse lung tissue, add 600 μL lysis solution RLTPlus and homogenize for 15 seconds; add all the lysate solution to the DNA removal column; centrifuge and retain the filtrate; add an equal volume of 70% ethanol and mix immediately by blowing; add the mixture to the adsorption column RA, centrifuge, and discard the waste liquid; add 500 μL of deproteinization solution RW1, stand at room temperature for 2 minutes, centrifuge, and discard the waste liquid; add 500 μL of rinse solution RW, centrifuge, and discard the waste liquid. Add 500 μL of rinse solution RW and repeat; put the adsorption column RA back into the empty collection tube and centrifuge; take out the adsorption column RA and put it into a clean RNase Free centrifuge tube, add 50 μL RNase Free H2O to the middle part of the adsorption membrane, stand at room temperature for 1 minute, centrifuge, and obtain total RNA, which is stored at -80℃.

[0098] According to the instructions of the H1N1 RealTime RTPCR kit, the reaction system mixture was prepared, PCR amplification reaction was performed, and FAM and HEX / VIC / JOE channels were used for detection.

[0099] The results are shown in Table 3.

[0100] Table 3 Group Viral load (copies / mL) Normal control group 124 Model control group <![CDATA[5.82×10 7 ]]> Tamiflu control group 8426 Example 1 5582 Example 2 5395 Example 3 5214 Comparative Example 1 7952 Comparative Example 2 8015 Comparative Example 3 8224 Comparative Example 4 10256 Comparative Example 5 13251 Comparative Example 6 25733 Comparative Example 7 12156

[0101] It can be seen from the above table that the Abidor Hydrochloride Sustained-Release Tablets prepared in Examples 1-3 of the present invention can significantly reduce the lung viral load of mice.

[0102] 3. Detection of NK cell activity in spleen immune cells

[0103] Mouse spleen was ground into single cell suspension, and red blood cell lysis solution was added to lyse red blood cells. BV510-CD3, PE-Cy7-NK1.1, and PE-NKG2D antibodies were added to the cells, incubated at 4°C in the dark for 20 minutes, centrifuged, and washed twice. 200μL Fluxation and Permeabilization Solution was added to the cells, incubated at 4°C in the dark for 30 minutes, centrifuged, and washed twice; FITC-granzyme A antibody was added, incubated at 4°C in the dark for 30 minutes; centrifuged, and washed twice. The activity of NK cells was detected by flow cytometry. The results are shown in Table 4.

[0104] Table 4 Group NK cell ratio (%) Normal control group 5.38±1.05 Model control group 14.92±3.56* Tamiflu control group 10.42±2.21# Example 1 12.21±2.05# Example 2 12.44±2.15# Example 3 12.57±2.34# Comparative Example 1 10.15±2.96 Comparative Example 2 10.46±2.78 Comparative Example 3 10.01±2.85 Comparative Example 4 11.59±2.91 Comparative Example 5 11.05±2.87 Comparative Example 6 9.68±2.65 Comparative Example 7 10.27±2.59 Note: * compared with the normal control group, P < 0.05; # compared with the model control group, P < 0.05.

[0105] It can be seen from the above table that the Abidor Hydrochloride Sustained-Release Tablets prepared in Examples 1-3 of the present invention can significantly increase the proportion of NK cells in the spleen immune cells of mice, promote the activation of NK cells, enhance the innate immune response, and accelerate the clearance of viral infection.

[0106] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A method for preparing arbidol hydrochloride sustained-release tablets, characterized in that: Licorice, ginger and kudzu flower are subjected to water extraction and alcohol precipitation to obtain hydrophilic active polysaccharides, and the active components are separated by ionic liquids. Abidor hydrochloride, biological toxins and active components are mixed to prepare liposomes, which are evenly mixed with lactose, cross-linked polyvinylpyrrolidone and hydrophilic active polysaccharides, and are prepared into rapid-release granules with polyvinylpyrrolidone K30 solution and magnesium stearate; Abidor hydrochloride and microcrystalline cellulose are mixed, and sustained-release granules are prepared with ethanol and magnesium stearate; the rapid-release granules and sustained-release granules are layered and tableted to prepare Abidor hydrochloride sustained-release tablets.

2. The preparation method according to claim 1, characterized in that: The following steps are involved: S1. Extraction of hydrophilic active polysaccharides: Licorice, ginger and kudzu flower are washed, dried and crushed respectively, added into water, heated to boiling for extraction, filtered, the solid residue is retained, ethanol is added into the liquid for precipitation, filtered, the filtrate is retained, the solid is washed and dried to obtain the hydrophilic active polysaccharide; S2. Extraction of active components: The solid residue in step S1 and the filtrate in step S1 are mixed and added to the ionic liquid, carbon dioxide is introduced, stirred for extraction, filtered, the filtrate is heated to allow carbon dioxide to escape, separated, the organic layer is collected, the active component is obtained, and the ionic liquid layer is washed and reused; S3. Preparation of liposomes: Abidol hydrochloride, biotoxins, and active ingredients were mixed uniformly, lecithin and glyceryl monostearate were added, and the mixture was stirred and dispersed uniformly to obtain a mixture; an aqueous solution containing polysorbate-80 and polyethylene glycol 400 was prepared, the mixture was added dropwise with stirring, ultrasonically dispersed uniformly, and freeze-dried to obtain liposomes; S4. Preparation of immediate-release granules: liposomes, lactose, cross-linked polyvinylpyrrolidone, and hydrophilic active polysaccharide are uniformly mixed, a soft material is prepared with a polyvinylpyrrolidone K30 solution, granulated, dried, granulated, and magnesium stearate is added and mixed to prepare immediate-release granules; S5. Preparation of sustained-release granules: Abidol hydrochloride and microcrystalline cellulose were mixed and sieved, ethanol was added to prepare a soft material, granulated, dried, granulated, magnesium stearate was added and mixed to prepare sustained-release granules; S6. Preparation of Abidor Hydrochloride Sustained-Release Tablets: The immediate-release granules and the sustained-release granules are layered and tableted in proportion to prepare Abidor Hydrochloride Sustained-Release Tablets.

3. The preparation method according to claim 2, characterized in that: In step S1, the mass ratio of licorice, ginger and kudzu flower is 3-5:5-7:10-15, and the time of heating to boiling extraction is 3-5h.

4. The preparation method according to claim 2, characterized in that: In step S2, the ionic liquid is selected from at least one of 1-butyl-3-methylimidazolium hexafluorophosphate, 1-butyl-3-methylimidazolium bistrifluoromethanesulfonyl imide salt, 1-butyl-3-methylimidazolium hexafluoroantimonate, 1-hexyl-3-methylimidazolium tetrafluoroborate, 1-hexyl-3-methylimidazolium hexafluorophosphate, and 1-hexyl-3-methylimidazolium hexafluoroantimonate, the carbon dioxide ventilation rate is 10-20 mL / min, the stirring extraction time is 20-30 min, and the filtrate heating temperature is 50-60 ° C.

5. The preparation method according to claim 2, characterized in that: The biotoxin in step S3 is selected from at least one of bee venom peptide, phospholipase A2, and bee venom peptide, the mass ratio of arbidol hydrochloride, biotoxin, active ingredient, lecithin and glyceryl monostearate is 15-20:3-5:4-6:25-35:100-150, and the concentration of polysorbate-80 in the aqueous solution containing polysorbate-80 and polyethylene glycol 400 is 0.5-1wt%, and the concentration of polyethylene glycol 400 is 1-2wt%.

6. The preparation method according to claim 2, characterized in that: The mass ratio of the liposome, lactose, cross-linked polyvinylpyrrolidone, hydrophilic active polysaccharide, polyvinylpyrrolidone K30 solution and magnesium stearate in step S4 is 10-15:4-7:15-20:3-5:30-50:0.5-1, and the concentration of the polyvinylpyrrolidone K30 solution is 5-10wt%.

7. The preparation method according to claim 2, characterized in that: The mass ratio of arbidol hydrochloride, microcrystalline cellulose, ethanol and magnesium stearate in step S5 is 8-10:4-7:15-20:0.5-1, and the mesh number of the sieve is 80-100 mesh.

8. The preparation method according to claim 2, characterized in that: The mass ratio of the rapid-release granules to the sustained-release granules in step S6 is 3-5:2-4.

9. An arbidol hydrochloride sustained-release tablet prepared by the preparation method according to any one of claims 1 to 8.

10. Use of the arbidol hydrochloride sustained-release tablets as claimed in claim 9 in the preparation of medicines for treating pneumonia and influenza.

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