KR0137206B1 - Spray gel base and spray gel preparation thereof - Google Patents

Abstract

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Description

Spray gel base and spray gel using the same

The present invention relates to a gel preparation suitable for spraying (hereinafter referred to as gel base for spraying) and a gel preparation suitable for spraying prepared by uniformly mixing the above-mentioned spraying gel base with an active drug (hereinafter for spraying Gel). More specifically, an electrodepositionable gel base having excellent electrodeposition properties, prepared by increasing the viscosity of an aqueous solution of a carboxyvinyl polymer (hereinafter referred to as CVP) with a water-soluble basic substance and uniformly dispensing the active drug thereon. It relates to a gel for spraying by mixing.

Sprays, such as aerosols and the like, using a fluorocarbon fluoride (i.e., Freon, a product of DuPont) as the propellant, and an active drug aqueous solution by manual pressurization, and the like are now known. Among these, aerosols using fluorinated hydrocarbons (freons) as propellants; (1) The sprayed active drug or powder containing the active drug must dissolve at the spray site to exert its pharmacological action, but it does not dissolve well, so the aerosol is inferior to the nebulizer of the active drug solution in order to maximize the pharmacological action. (2) furthermore, there is a physical irritation at the spray site due to the fluorinated hydrocarbons themselves and the gas atomization pressure, and (3) and because the fluorinated hydrocarbons have a severe effect on the ozone content in the stratosphere, It is not preferable because it becomes.

On the other hand, there are a number of other drawbacks, although the nebulizer for spraying an aqueous solution of the active drug by manual press does not have the drawbacks mentioned above in the aerosol. In other words, the spray of the active drug solution has poor electrodeposition properties, so the active drug solution flows down from the spray site, resulting in anxiety when used, and the active drug cannot be administered to the fixed site. If not dissolved, the formulation containing the active drug cannot be prepared uniformly.

Under the above circumstances, for example, attempts have been made to prevent the liquid droplets from dropping by some means such as minimizing the size of the spray nozzle of the sprayer to make the particle size much smaller when spraying. However, the problem of the liquid drop still being solved by such means has not been solved. Therefore, studies have been conducted to develop a means for maintaining the required amount of the active drug properly at the spray site without the liquid drop flowing.

In order to improve the electrodeposition property of the spray agent which is an aqueous solution of the active drug at the time of spraying, a water-soluble high molecular compound generally used as a thickener, for example, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, polyvinyl alcohol , Polyvinyl pyrrolidone, gelatin, sodium alginate and the like are effective in increasing the viscosity of the aqueous active drug solution. However, according to the researches of the present inventors, when using these conventional thickeners, the aqueous solution of the active drug cannot be sprayed from the sprayer, or even if it can be sprayed, the sprayed liquid does not become a misty form and becomes a water column. It remains unresolved as such a means.

In a thorough study of spray formulations, the inventors found that gel bases prepared by increasing the viscosity of a CVP aqueous solution with water-soluble basic materials have good sprayability in the sprayer, whereas the gel bases prepared in this way employ the conventional thickeners described above. It has been found that it has a high viscosity compared to the solution prepared in the above, and that the dripping of the liquid can also be prevented by using the gel base described above except in the case of application to a living body. However, the present inventors have found the following drawbacks in the above-described gel base made from CVP. That is, the viscosity of the high CVP solution before spraying is somewhat lowered by spraying, and when applied to a living body such as mucous membranes or skin, the viscosity of the solution decreases rapidly at the spraying site, and the liquid drips or contains the required amount of active drug. It can be maintained properly. In order to solve the above-mentioned drawbacks, the present inventors attempted to increase the viscosity of the active drug solution by using CVP at a high rate, but in this case, high spraying pressure was required to spray it, and high spraying cancer at the application site. There was a stimulus.

Moreover, forced spraying of such high viscosity solutions results in very large particle sizes of the solution,

A higher viscosity of the solution makes it impossible to spray.

Under the circumstances described above, the inventors have made more and more intense studies and, as a result, the aqueous solution containing a relatively high concentration of CVP is increased to a water-soluble base material to obtain a gel having a relatively high viscosity, and then the viscosity is increased. It was unexpectedly found that by adjusting the range to 500-5,000 centipoise (cp) as a modifier, a useful spray gel base with good properties could be obtained. That is, it was unexpectedly found that the gel base for spraying prepared by the above-described method was able to obtain the required gel base for spraying having a small viscosity change and excellent properties between before and after spraying. In other words, the gel base for spraying prepared by the above-described method has a small viscosity change and excellent electrodeposition property between before and after spraying, so that when applied to a living body such as mucous membrane or skin, it does not flow from the application site. Furthermore, the present inventors have found that spray gels prepared by mixing the active drug with the gel bases described above also have very good properties and can continue to release the active drug against living organisms such as mucous membranes and skin.

It is an object of the present invention to increase the viscosity of a relatively high concentration of aqueous solution containing CVP with a water-soluble basic material, and then to adjust the viscosity of the thickened solution to a certain range as a viscosity modifier to prepare a good electrodeposition and spray-free spray gel in vivo application It provides a mechanism. Another object of the present invention to provide a spray gel prepared by uniformly mixing the active drug and the above-described gel base with an active drug having excellent absorption of the active drug when applied to a living body. Various objects and advantages of the present invention will become apparent from the following description.

The present invention provides a CVP of 0.2-1.5 wt. The aqueous solution containing% was used to increase the viscosity with a water-soluble base material, and the viscosity was adjusted within the range of 500-5,000 cp with a viscosity modifier so that the spray particle size distribution after spraying was 80% or more in the range of 20-100 μl. Provided is a spray gel base having good electrodeposition properties comprising an active drug and a spray gel base.

Hereinafter, the present invention will be described in more detail.

The CVP used in the spray gel base of the present invention is a hydrophilic polymer prepared by synthesizing acrylic acid as the main monomer component, and is manufactured by conventional Carbopol 934,934P, 940 and 941 [commercially available from Goorich, USA. )] And the like. The concentration of the CVP aqueous solution used in the present invention is generally 0.2-1.5 wt. It is in the range of%.

The water-soluble base material used in the present invention is used for the purpose of thickening the CVP aqueous solution to increase its viscosity. Suitable examples of the water-soluble base material of the present invention include inorganic bases (e.g. sodium hydroxide, potassium hydroxide, ammonia, etc.) and organic bases [e.g. alkalamines (e.g. methylamine, ethylamine, propylamine, etc.), dialkylamines (e.g. : Dimethylamine, diethylamine, dipropylamine, etc.), trialkylamines (such as trimethylamine, triethylamine, tripropylamine, etc.), alkanolamines (such as methanolamine, ethanolamine, propanolamine, etc.), Dialkanolamines (e.g. dimethanolamine, diethanolamine, dipropanolamine, etc.), trialkanolamines (e.g. trimethanolamine, triethanolamine, tripropanolamine, etc.), amino acids (e.g. arginine, lysine, ornithine, etc.) ), Etc.]. These water soluble bases are used in amounts necessary for neutralization to adjust the pH value of the CVP aqueous solution to the desired pH value.

The viscosity modifier of the present invention is a CVP 0.2-1.5 wt. It is used for the purpose of adjusting the viscosity of a relatively high viscosity gel prepared by thickening the aqueous solution containing% with a water-soluble base material so that the spray particle size distribution after spraying is 80% or more in the range of 20 to 100 µm. Suitable viscosity modifiers of the invention include, for example, sodium chloride, potassium chloride, calcium chloride and the like. The viscosity modifier of the present invention is 0.01-10.2 wt.% With respect to the total amount of all components. It is preferable to use it in the ratio of%. On the other hand, when applied to the mucosa, the amount of viscosity modifier should be determined in consideration of the change in osmotic pressure due to the viscosity modifier.

In adjusting the viscosity of the gel base for spraying of this invention, it is preferable to make the spray particle size distribution after spraying be 80% or more in the range of 20-100 micrometers. Only when the spray particle size distribution after spraying is in the said range, the spray gel base of this invention has the outstanding electrodeposition property, and the viscosity does not change before and behind spraying.

The spray gel base of the present invention contains 0.2-1.5 wt. It can be prepared by adding a water-soluble base material to the aqueous solution containing%, uniformly mixing the mixture to prepare a viscous gel, and then adding a viscosity modifier under stirring to obtain the required viscosity. When a viscosity modifier is a crystalline form, it can also add as it is, but it is more preferable to add in the form of aqueous solution. This is because when added in the form of an aqueous solution, there is no sudden change in viscosity and the viscosity changes uniformly.

The pH value of the spray gel base of the present invention is adjusted with a water soluble base material or other pH adjuster taking into account the stability or absorbency of the active drug.

The spray gel of the present invention is prepared by thickening an aqueous solution containing 0.2-1.5 wt.% CVP with a water-soluble base material, then uniformly mixing the active drug and adjusting the viscosity of the mixture in the same manner as the spray gel base described above. do. Furthermore, according to the type of active drug, the spray gel of the present invention first dissolves or disperses the active drug in an aqueous solution containing 0.2-1.5 wt.% Of CVP, and then uniformly mixes the same by adding a water-soluble base material under stirring. It is prepared by adjusting the viscosity of the mixture by the method.

Both water-soluble and water-insoluble drugs can be used as the active drugs of the present invention, but it is more preferable to use drugs which are stable in the preparation, ie in aqueous solution. Examples of suitable active drugs of the present invention include hypnotics and sedatives (e.g., glutetimides, chloral hydrates, nitrazepam, amobarbital, phenobarbital, etc.), antipyretic, analgesic and anti-inflammatory agents (e.g. aspirin, acetaminophen, ibuprofen, flur) Bipropene, Indomethacin, Ketoprofen, Thiclofenac Sodium, Thialamide, Pyroxycane, Flufenacic Acid, Mefenamic Acid, Pentazosin, etc., Local Anesthetics (e.g. Methyl Aminobenzoate, Lidocaine, etc.) ), Local vasoconstrictors (e.g. napazoline nitrate, tetrazoline nitrate, oxymetazone hydrochloride, tramizoline hydrochloride, etc.), anti-allergic agents (e.g. disodium chromoglycolate, oxatomid, azelastine hydrochloride, fructose Ketoxifene, sodium traxanox, amlexanox, etc.), cardiac agents (e.g., dopamine hydrochloride, ubidecarenone, etc.), antiarrhythmic agents (e.g., propranolol, pindrol, phenytoin, disopyramid, etc.), coronary blood Dilators (e.g. isosorbide nitrate, nifedipine, diltiazem hydrochloride, dipyridamole, etc.), drugs of the digestive system (e.g. domperidone, etc.), corticosteroids (e.g. triamcinolone acetonide, dexamethasone, betametha Zonsodium phosphate, predonisolone acetate, fluorinide, fluoropropionate beclomethasone, flunisolide, etc., antipramine (e.g., tranexamic acid), antifungal agents (e.g. clotrimazole, nitrate myconazole, ketoconazole Etc.), anti-malignant neoplastic agents (e.g. tegapur, fluorouracil, mercaptopurine, etc.), antibiotics (e.g. amoxicillin, ampicillin, cephalexin, cephalotin sodium, ceftizone sodium, erythromycin, hydrochloric acid) Oxytetracycline, etc.), bioactive peptides (e.g., calcitonins such as insulin, salmon calcitonin, chick calcitonin and elcatonin, urokinase, TRA, inteferon, etc.), waxes (e.g., influenza wax, swine) Reude include those such telra infection prevention confirmed, B-type infection wakjin etc.). The amount of active drug used in the spray gel of the present invention varies depending on the type of drug, but the active drug is used in an amount sufficient to exert a useful pharmacological activity.

In the present invention, when the water-insoluble drug is used, the spray gel becomes cloudy in white, but the active drug does not settle, and there is no problem in daily administration. However, when the spray gel of the present invention is applied to a skin or the like and the absorption into the living body is better in a solid form than the solid, it is preferable to dissolve the water-insoluble drug in a water-soluble organic solvent or prepare a spray gel using a dissolving agent. Do. Suitable water soluble organic solvents include, for example, lower alcohols (eg ethanol, isopropanol, etc.), glycols (eg propylene glycol, 1,3-butylene glycol, polyethylene glycol having a molecular weight of 300-500, etc.). Suitable solubilizers are selected from the group consisting of various surfactants, crotamiton, salicylate glycol esters, methyl salicylate, peppermint oil, benzyl alcohol, and the like, depending on the solubility of the active drug.

On the other hand, the active drug used in the present invention can be suspended by using a suitable suspending agent. Suitable suspending agents include various surfactants, such as sucrose fatty acid esters, polyoxyl stearate 40, polyoxyethylene hydrogenated castor oil 60, polysorbate 80, glycerin monostearate, voicebita monostearate, sorbitan Monostearate, sorbitan monopalmitate and the like.

It is preferable to adjust the viscosity of the spray gel base and the spray gel of the present invention within the range of 500-5,000 cps with a viscosity modifier such as sodium chloride, potassium chloride or calcium chloride. When the viscosity of the spray gel base or spray gel of the present invention is 500 cps or less, the fluidity is too high, and when applied to mucous membranes or skin, the liquid flows down. On the other hand, when the viscosity of the spray base or spray gel of the present invention is 5,000 cps or more, the particle size after spraying is irregular and large, which is not suitable for showing the desired effect of the active drug well. More preferably, the viscosity of the spray base or spray gel of the present invention is within the range of 800-3,000 cp.

Spray gel of the present invention can be applied to the nasal mucosa, oral mucosa, vaginal mucosa and skin according to the conventional method. Compared with gels prepared without the use of viscosity modifiers or with CVP gel bases or formulations prepared using conventional water-soluble polymer compounds, the spray gel bases and spray gels of the present invention have a much more uniform particle size and change in viscosity between before and after spraying. Because of the small amount, the electrodepositability is excellent and the liquid does not flow down after spraying.

Moreover, the spray gel of the present invention is also useful for medical use. For example, spray gels of influenza wax prepared according to the present invention can be applied to the nasal mucosa and are more preferred than the dosage forms of conventional influenza waxes.

Until now, influenza waxes have been administered by subcutaneous injection, because influenza waxes are high molecular weight polypeptides, which have poor membrane permeability, tend to degrade in the digestive tract, are difficult to be absorbed by living bodies, and moreover, influenza when administered orally This is because the inoculation of Luenzer wax is aimed at the production of antimicrobial antibodies in the blood (fluid immunity).

Essentially, influenza bacteria infect only the mucous membrane of the respiratory tract (local infection), so it is more effective to produce antibodies (I gA antibodies) on the mucous membrane of the respiratory tract than to produce antibodies in the blood. However, in conventional subcutaneous inoculation, secretion I gA can be obtained since the mucous membrane of the respiratory tract is not irritated. In addition, subcutaneous vaccination is limited in use due to its side effects, and the antibody response level is not appropriate and the duration is short, because a sufficient amount of influenza wax cannot be inoculated to prevent infection.

In addition, because influenza bacteria are prevalent as they cause fragrance displacement, if the types of antigens in waxes are different from those of prevalent bacteria, the prevention effect of waxes is reduced.

In order to ameliorate the above drawbacks of conventional influenza wax dosage forms, efforts have been made to develop new types of waxes or dosage forms thereof. Administration into the nasal cavity (nasal cavity) may induce secretory I gA antibodies in the respiratory tract, one infectious route and is effective in preventing infection.

However, it is difficult to obtain high levels of secreted I gA in the respiratory tract by spraying the influenza wax alone or by spraying the influenza wax aqueous solution. The reason is that because influenza wax is a high molecular weight polypeptide, it has low permeability in the respiratory tract mucosa, and in respiratory tract mucosa due to the purifying mechanism of the respiratory tract mucosa such as mucus secretion, epithelial wear, villius movement, etc. This is because absorption of influenza wax is difficult.

The nasal spray gel of the influenza wax of the present invention may be prepared by mixing the spray gel base and the influenza wax described above. The influenza wax used in the present invention may be either a toxic weakened wax or an inactive wax thereof.

When using toxic weakened wax, it may be H _A wax made by removing fat component by ether treatment or viral particle wax without ether treatment.

Moreover, influenza waxes used in the present invention include those such as new cold-adapted fresh waxes, artificial waxes, genetically modified waxes, and peptide waxes.

Nasal spray gels of influenza waxes prepared according to the present invention may contain suitable active drugs, fungicides, preservatives, surfactants, stabilizers and the like that can be used with influenza waxes.

The present invention will be described in more detail in the following examples with reference examples and experimental examples, but should not be construed as being limited thereto. In the following experimental example, the reference example, and the Example, the viscosity was measured with C-type viscosity meter [made by Tokyo Keiki Co., Ltd. of Japan] at 20 degreeC.

Experimental Example

The spray test was carried out on various bases prepared using the following various thickeners and purified water, and the properties of the bases were measured by spray state, viscosity retention, electrodeposition, and skin electrodeposition. The results are shown in Table 1.

TABLE 1

HPMC: hydroxypropyl methylcellulose

HPC: Hydroxypropyl Cellulose

PVA: Polyvinyl Alcohol

PVP: Polyvinyl Pyrrolidone

*) Spray condition was evaluated according to the following criteria.

Poor 1: No ejection from sprayer

Poor 2: The solution from the sprayer was not a particle but a column of water.

Poor 3: Ejected from sprayer, but too large particles.

Good: Uniformly ejected from the sprayer, small particles.

**) Viscosity retention was calculated according to the following equation.

***) The electrodeposition test was carried out in the following manner.

Filter paper No. 6 (diameter: 110 mm) infiltrated with saline solution (1.5 g) was attached to an inclined plate at an angle of 40 degrees, and then sprayed the contents of the sprayer (600 mg) toward the center of the filter paper at a distance of 30 mm, and then the liquid The time (seconds) until starting to flow was measured. When the liquid did not flow out, it was evaluated as no change.

****) Skin electrodeposition was evaluated by the following method.

After spraying the contents of the sprayer (180mg) inside the upper forearm of a person at a distance of 30mm, if the liquid spilled within 10 seconds, it was regarded as dripping, and if the liquid did not flow for 10 seconds after spraying, Evaluated by

As is clear from Table 1, only the gel base prepared according to the present invention (CVP 0.4% + NaCl 0.27% and CVP 0.6 + NaCl 0.45%) has all spraying state, viscosity retention, electrodeposition to plate and human skin. It was good.

Example 1

Gel gel for spraying of ketoprophen:

Ketoprofen spray gel was prepared using the following amounts of ingredients.

To the CVP 4% aqueous solution was slowly added an aqueous 2% sodium hydroxide solution while stirring and the mixture was stirred until it became a gel. A 1% aqueous solution of disodiumedetate was added to the mixture, followed by the slow addition of a suspension in which ketoprofen was suspended in polysorbate 80 and purified water, and then stirred uniformly. Kerosene spray gel (3%, pH: 6.8, viscosity: 3,800 cps) was prepared by adjusting the viscosity of this mixture with an aqueous 10% sodium chloride solution and then stirring the mixture uniformly and mixing well.

Example 2

Gel for spraying nitrate tetrazoline:

Gel for spray tetranitrate nitrate was prepared using the following amounts of ingredients.

To the CVP 4% aqueous solution was slowly added a 2% aqueous solution of L-arginine, and the mixture was stirred until it became a gel. Tetrazoline nitrate dissolved in purified water was slowly added and the mixture was stirred uniformly, and then the viscosity of the mixture was adjusted to 10% aqueous sodium chloride solution, and the mixture was stirred and mixed uniformly to spray gel of tetrazoline nitrate (0.1 %, pH: 5.8, viscosity: 4,500 cps).

Example 3

Gel for spraying disodium chromoglycolate:

Spray gels of disodium chromoglycolate were prepared using the following amounts of ingredients.

2% aqueous sodium hydroxide 2% solution was slowly added to the CVP 4% aqueous solution while stirring, and the mixture was stirred until it became a gel. A 1% aqueous solution of disodium edetate was added to the mixture, and a solution obtained by adding sodium chromoglycolate to glycerin and purified water was added and stirred uniformly. The viscosity of this mixture was adjusted to 10% sodium chloride, 10% aqueous solution, and uniformly stirred and mixed to prepare a gel for spraying disodium chromoglycolate (2%, pH: 6.0, viscosity: 1500cp).

Example 4

Oxatomide spray gel:

A spray gel of oxatomid was prepared using the following amounts of ingredients.

To the CVP 4% aqueous solution, an aqueous 2% aqueous solution of L-arginine was slowly added while stirring until the mixture became a gel. The mixture was added to polysorbate 80 and purified water, and the suspension obtained by adding oxatomid was gradually added and stirred uniformly. The viscosity of the mixture was adjusted with an aqueous 10% sodium chloride solution and uniformly stirred to prepare a gel for spraying oxatomide (0.01%, pH: 5.1, viscosity: 1500 cps).

Example 5

Gel for spraying propionic acid beclomethasone:

Spray gel of propionic acid beclomethasone was prepared using the following amount of components.

Slowly add 2% aqueous sodium hydroxide solution to the CVP 4% aqueous solution, stirring until the mixture becomes a gel.

Stirred. Slowly add polysorbate 80, concentrated glycerin and beclometazone propionate to purified water

The suspension was slowly added and stirred uniformly.

The viscosity of this mixture was adjusted to 10% aqueous sodium chloride solution, uniformly stirred and mixed, and propionate beclometa.

Zone spray gel (0.1%, pH: 6.0, viscosity: 2500 cps) was prepared.

Example 6

Polyniside Spray Gel:

Polynilide's spray gel was prepared using the following amounts of ingredients.

While stirring to a 4% aqueous solution of CVP, an aqueous 2% sodium hydroxide solution was slowly added and stirred until the mixture became a gel. A 1% aqueous solution of disodium edetate and 0.1% aqueous solution of belsalkonium chloride were added to the mixture, followed by the addition of polysorbate 80, polyglycolide to polyethylene glycol and purified water, and agitation.

The viscosity of the mixture was adjusted to a 10% aqueous solution of sodium chloride and the mixture was stirred and mixed uniformly to prepare polynisole's spray gel (0.0255%, pH: 5.1, viscosity: 2,200 cps).

Example 7

Gel for spraying insulin:

Spray gels of insulin were prepared using the following amounts of ingredients.

L-arginine 4% aqueous solution was added slowly, stirring to CVP 4% aqueous solution, and this mixture was stirred until it became a gel. To this mixture, a solution obtained by adding insulin to purified water was slowly added and stirred uniformly. The viscosity of this mixture was adjusted with an aqueous 10% sodium chloride solution and uniformly stirred to prepare a spray gel of insulin (50 U / g pH: 7.3, viscosity: 550 cps).

Example 8

Gel for nasal spraying of influenza H _A waxes:

Nasal spray gels of influenza H _A waxes were prepared using the following amounts of ingredients.

L-argin 4% aqueous solution was slowly added to the CVP 4% aqueous solution while stirring, and the mixture was stirred until it became a gel. To the mixture was slowly added a solution obtained by adding influenza H _A wax to the phosphate buffer and stirred uniformly.

The viscosity of this mixture was adjusted to a solution obtained by adding sodium chloride to purified water, and the mixture was uniformly stirred and mixed to obtain a nasal spray gel of influenza H _A wax (249 μg protein / ml, pH: 7.2, viscosity: 2,900 cps). Was prepared.

Example 9-11

Influenza H _A wax (A / Yamagata / 120/86) Nasal spray gels were prepared using the amounts of ingredients in the following table.

TABLE 2

*) A / Yamagata / 120/86, 500µg Protein / ml

To the CVP 4% aqueous solution was slowly added an aqueous 8% aqueous solution of L-arginine and stirred until the mixture became a gel. A solution obtained by adding sodium chloride to purified water was slowly added to the mixture, followed by uniformly stirring to prepare a base for spraying. The spray gel base thus prepared was subjected to high pressure steam sterilization (121 ° C., 20 minutes), and then uniformly mixed with influenza H _A wax solution (500 μg protein / ml) and concentrated under aseptic conditions to influenza H _A. Wax nasal spray gels were prepared.

Reference Example 1

Influenza Me H _A wakjin (A / Yamagata / 120/86), a phosphate buffer solution (pH: 7.4) influenza that H _A was dissolved in the wax spirit nasal

Spray formulations (24 μg protein / ml) were prepared.

Reference Example 2

Influenza H _A wax (A / Yamagata / 120/86) was dissolved in phosphate buffer (pH: 7.3) to prepare a nasal spray formulation (250 μg protein / ml) of influenza H _A wax.

Pharmacological test

The following pharmacological test was carried out using the nasal spray gel of the influenza H _A waxes prepared in Examples (8 to 11) and the nasal spray formulations of the influenza H _A waxes prepared in Reference Examples 1 and 2.

(Way)

(1) Immunology:

Intraperitoneal injection of Nembutal injection (sodium fetobarbiturate, 15-fold diluent) into BALB / C mice

(0.2-0.3 ml / mar) and anesthetized.

The nasal passages of anesthetized BALB / C mice were turned upward and gelled (20 μg) into one of the nasal passages.

(2) Sampling Method:

At 3 weeks post-immunization, blood serum was collected (whole blood) for measurement of hemagglutination inhibitory antibody value (hereinafter, simply referred to as HI antibody value), and the thoracic incision was exposed to trachea. Dilution of anti-IgA antibody (1 ml) was injected into the trachea from the top of the trachea under the jaw toward the lungs, and was slowly absorbed. Samples were taken by repeating this suction and injection.

(3) HI antibody value measurement method:

Serum samples (0.1 mL) were treated with receptor disrupting enzyme (RED) (neuuraminidase, 0.3 mL), kept at 37 ° C. overnight, followed by passivation at 56 ° C. for 1 hour. One drop of chicken red blood cells was added thereto, and the mixture was shaken and left at room temperature for 1 hour, after which the supernatant was collected in a centrifuge (2,000 rpm, 10 minutes). The antigen amount of the strain used for the measurement was adjusted to 16 H _A. Phosphate buffer (0.025 mL) was injected into the whole, except for the first well above the 96 wells of the microplate. In the first well a sample (0.05 ml) was placed. Samples in all wells were diluted with an automatic dilutor. The amount of antigen solution (0.025 ml) steered to 16 H _A was injected into all wells of the microplate, and the plate was shaken with a plate mixer for 1 hour. 0.5% of blood cells (0.05 ml) was added to all wells, then the plate was shaken again, left for 1 hour and the shape of each well bottom was observed.

(4) IgA test method:

HANA antigen was diluted to 0.05 M carbonate buffer (pH: 9.5) using Dynatech Immulon II Flatbottom Plates to obtain an optimal concentration (approximately 1-10 μg / ml). The wells were injected into each well of the plates described above and left overnight at 4 ° C. (about 18 hours). The next day, the plates were washed three times with 0.01 M phosphate buffer (pH: 7.2) containing 0.05% Tween 20 and then treated at 37 ° C. for 1 hour with phosphate buffer containing 0.1% BSA (bovine serum albumin). Blocking made the antigen solids. A control plate was prepared in the same manner as above except that serum serum solution was used instead of HANA antigen.

100 μl of dilution for the sample (0.01 M phosphate buffer containing 0.5% BSA and 0.05% Tween 20) was added.

This plate was wrapped with Saran Wrap (polyvinylidene chloride film) and then left at 4 ° C. overnight. The next day, the reaction mixture in each well was removed and each well was washed three times with a wash (0.01 M phosphate buffer containing 0.05% Tween 20).

Labeled antibodies (anti mouse IgA antibodies labeled with peroxidase) were diluted to optimal concentration with the same dilution for such samples and 100 μl was injected into the wells of the plate.

The plate was allowed to react at room temperature for 2 hours, and then washed three times with the above-mentioned washing liquid.

100 µl of substrate solution (0.1 M citric acid buffer, pH: 4.9, containing 3.3 mg / ml 0-phenylenediamine, 0.02% H202) was injected into each well, and reacted for 0.5-1.0 hours at room temperature under shaded conditions. N sulfuric acid (100 µl) was added to stop the reaction. Absorbance (492 nm) was measured by autoreader on a microplate.

(5) result:

The results are shown in the following third table.

TABLE 3

As is evident from the above results, in mice administered the nasal spray gel of the influenza H _A wax prepared according to the present invention, antiviral IgA antibody was detected in the nasal lavage three weeks after administration, and also the HI antibody. Hyperemia concentrations were detected at high levels. In contrast, traces of IgA antibodies were detected in the mice to which the influenza H _A wax was added to the phosphate buffer (Reference Examples 1 and 2 degrees) and blood HI antibodies were detected only at low levels under the same conditions.

Claims (12)

0.2-1.5 wt. Of aqueous carboxyvinyl polymer solution. Gel base for spraying containing aqueous solution of carboxyvinyl polymer of viscosity 500-5,000 centipoise prepared by adjusting the viscosity to 500-5,000 centipoise after adjusting the viscosity to a viscosity modifier. The gel base for spraying according to claim 1 wherein the viscosity modifier is selected from the group consisting of sodium chloride, potassium chloride and calcium chloride. The spray gel base of claim 1 wherein the pH value is in the range 4-9. The gel base for spraying according to claim 2 wherein the pH value is in the range 4-9. The gel base for spraying according to claim 1, 2, 3 or 4, wherein the water-soluble base material is a water-soluble inorganic base or a water-soluble organic base. A spray gel containing a homogeneous mixture of the spray gel base according to claim 1 and an effective amount of the active drug. The spray gel according to claim 6, wherein the pH value is in the range of 4-9. The gel for spraying according to claim 6 or 7 for applying to mucous membranes or skin. The gel for spraying according to claim 6 or 7, wherein the active drug is suspended by adding a dissolving agent or a solvent. The spray gel preparation according to claim 6 or 7, wherein the active drug is suspended by adding a suspending agent to the active drug. A gel agent for nasal spraying comprising the gel base according to claim 1 and an effective amount of influenza selected from the group consisting of viral particle wax, H _A wax, live virus wax, artificial membrane wax, genetically modified wax and peptide wax. The nasal spray gel of claim 11, wherein the pH ranges from 6.0-8.0.