An external spray containing itraconazole and eucalyptol and a method for preparing the same

Abstract

The application discloses a kind of containing it is the preparation method of spray and preparation method of eucalyptol for external use, and each 1000ml external use spray material composition: it is 8~10g, eucalyptol 10~20g, hydroxypropyl-β-cyclodextrin 250~300g, propylene glycol 100~140g, 9.5~12g of hydrochloric acid, diethylene glycol monoethyl ether 140~160g, caprylic acid capric acid polyethylene glycol glyceride 40~60g, 1.9~2.1g of sodium hydroxide, the rest is pure water, and preparation is obtained external use spray, mass is stable, antifungal effect is remarkable, clinical use is convenient.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of medicine, and particularly relates to an external spray containing itraconazole and eucalyptol and a preparation method thereof. BACKGROUND

[0002] Fungal skin infection is a common skin disease in dogs and cats, has strong transmission ability, and belongs to zoonosis. Skin fungal infection can be generally divided into two categories, superficial fungal infection and deep fungal infection, wherein deep fungal infection is more harmful but has a lower incidence; and superficial fungal infection is less harmful, but has strong infectivity and a high incidence, which brings many health risks to the sick dogs and cats and pet owners. In the clinical treatment of fungal skin disease in dogs and cats, the treatment is slow in effect, the treatment cycle is long, and the disease is prone to recurrence. In addition, the existing antifungal drugs are mostly in the form of oral administration, and there are problems of poor compliance and toxic side effects in the actual application.

[0003] Itraconazole is a triazole broad-spectrum antifungal drug, which is developed by Johnson & Johnson Company, USA, and can effectively treat fungal infections caused by Trichophyton, Microsporum, Epidermophyton floccosum, Candida, Blastomyces dermatitidis and the like. Itraconazole mainly affects the activity of cytochrome P450-dependent sterol 14α-demethylase of fungal cells, prevents the synthesis of ergosterol, an important component of fungal cell membranes, so as to inhibit fungal proliferation and promote fungal death.

[0004] Eucalyptol is a terpene, which widely exists in the oil of many eucalyptus trees, is a mixture of alpha-eucalyptol and beta-eucalyptol, and has certain antibacterial effect by destroying the cell wall and cell membrane of fungi, and can change the permeability, so that more drug molecules enter the fungal cells, and the combination of the two can improve the antifungal effect.

[0005] In the clinical treatment of pet skin fungal infection, there are very few drugs that can be selected, and most of them are oral preparations, such as ketoconazole tablets and itraconazole oral solution. In the medication treatment process, there are disadvantages such as large oral dose, long administration time, gastrointestinal stimulation (vomiting, anorexia, etc.) and liver and kidney toxicity. SUMMARY

[0006] In view of the above-mentioned deficiencies, the purpose of the present application is to provide an external spray containing itraconazole and eucalyptol which has significant antifungal effect and is convenient for clinical use, and a preparation method thereof.

[0007] To achieve the above-mentioned purpose, the present application adopts the following technical solutions:

[0008] An external spray containing itraconazole and eucalyptol, the material composition of each 1000ml of the external spray is: itraconazole 8~10g, eucalyptol 10~20g, hydroxypropyl-β-cyclodextrin 250~300g, propylene glycol 100~140g, hydrochloric acid 9.5~12g, diethylene glycol monoethyl ether 140~160g, caprylic capric acid polyethylene glycol glycerin ester 40~60g, sodium hydroxide 1.9~2.1g, and the rest is purified water; wherein the amount of purified water is used for preparing solution to 1000ml.

[0009] Further, the material composition of each 1000ml of the external spray is: itraconazole 9g, eucalyptol 15g, hydroxypropyl-β-cyclodextrin 270g, propylene glycol 120g, hydrochloric acid 10.5g, diethylene glycol monoethyl ether 150g, caprylic capric acid polyethylene glycol glycerin ester 50g, sodium hydroxide 2g, and the rest is purified water; wherein the amount of purified water is used for preparing solution to 1000ml.

[0010] Further, the preparation method of the external spray is specifically as follows:

[0011] 1) Take the prescription amount of 45% purified water, heat to 80℃, add the prescription amount of propylene glycol, 80℃, 50r / min stirring for 3min, to obtain solution 1;

[0012] 2) Slowly add the prescription amount of hydroxypropyl-β-cyclodextrin to solution 1, stir until completely dissolved, to obtain clear and transparent solution 2;

[0013] 3) Add the prescription amount of hydrochloric acid to solution 2, stir for 3min, then add the prescription amount of itraconazole, stir for 45~60min, to obtain clear solution 3;

[0014] 4) First, take the prescription amount of diethylene glycol monoethyl ether and caprylic capric acid polyethylene glycol glycerin ester, 50r / min stirring for 3min, then add the prescription amount of eucalyptol, stir for 10min, to obtain light brown solution 4;

[0015] 5) Slowly add the obtained light brown solution 4 to clear solution 3, stir for 20min, then add the prescription amount of sodium hydroxide, stir until completely dissolved, finally add the rest of the purified water to constant volume, stir for 30min, and pass through a 0.22μm microporous filter, then fill into a pressure container, to obtain the external spray containing itraconazole and eucalyptol.

[0016] By using the above scheme, the present application has the following advantages:

[0017] 1. This invention is based on the fact that both itraconazole and eucalyptol have good lipid solubility, and that itraconazole has the characteristic that its pharmacokinetics in the skin is delivered through sebum, which can make its concentration in the skin much higher than that in plasma. Therefore, the topical skin preparation developed has the advantages of significant antifungal effect and convenient clinical use.

[0018] 2. This invention prepares a topical spray by combining the broad-spectrum antifungal drugs itraconazole and eucalyptol. It is a novel formulation for treating superficial fungal infections in dogs and cats. It can be directly sprayed onto the lesion site in dogs and cats, firstly avoiding contact between pet owners and the infected lesion, thus reducing the risk of zoonotic diseases. Secondly, it avoids the gastrointestinal irritation, liver and kidney toxicity, and other side effects associated with oral administration. It also solves the problem of poor palatability of oral administration in dogs and cats, improving medication compliance. It is convenient to use in clinical practice, has excellent effects, is stable, and has a certain special fragrance.

[0019] 3. In this invention, itraconazole and eucalyptol have a synergistic effect. Combined with an innovative dosage form, the medication is administered to the lesion site, resulting in significant transdermal absorption and remarkable clinical efficacy. It is also convenient and safe to use. Furthermore, the preparation process is simple and low-cost, making it suitable for widespread application.

[0020] Other advantages, objectives, and features of the invention will be set forth in part in the description which follows, and in part will be apparent to those skilled in the art from the following examination, or may be learned from practice of the invention. The objectives and other advantages of the invention can be realized and obtained through the following description. Attached Figure Description

[0021] Figure 1 For itraconazole in vitro percutaneous permeation test Q -t curve;

[0022] Figure 2 For the in vitro transdermal permeation test of eucalyptol Q -t curve;

[0023] Figure 3 A comparison of clinical outcomes before and after treatment in Case 1;

[0024] Figure 4 This is a comparison of the clinical effects before and after treatment in Case 2. Detailed Implementation

[0025] The present invention will now be described in detail with reference to the accompanying drawings and embodiments. However, the embodiments do not limit the present invention in any way. Unless otherwise specified, the reagents, methods and equipment involved in the present invention are conventional reagents, methods and equipment in this technical field.

[0026] Example 1: Antibacterial test of itraconazole and eucalyptol

[0027] 1. Experimental method

[0028] According to the microdilution method of the American Clinical and Laboratory Standards Institute (CLSI), 15 strains of clinical isolates of Microsporum canis were selected for in vitro combined drug susceptibility testing, and the fractional inhibitory concentration index (FICI) was calculated. The FICI was used as the determination standard (when FICI≤0.5, it is synergistic; 0.5<FICI≤4.0, it is additive or irrelevant; when FICI>4.0, it is antagonistic). The drug interactions were analyzed, and Candida parapsilosis ATCC22019 was used as the quality control strain.

[0029] Preparation of drug solution: Itraconazole and eucalyptol were dissolved in dimethyl sulfoxide (DMSO) to prepare drug stock solutions of 320 μg / ml and 10 mg / ml, respectively, and stored frozen for future use. When used, the drug stock solutions were diluted with RPMI 1640 liquid medium to prepare working solutions with a final concentration of 2 times.

[0030] Bacterial liquid culture: The isolated test strains were cultured on SDA medium at 28°C for 7 days, and subcultured twice. After activation, the test bacteria were taken in a sterile test tube containing 2 ml of normal saline, allowed to stand, and the supernatant was shaken. The turbidimeter was used to adjust the concentration of the bacterial suspension to 5×10 6 CFU / ml. The RPMI 1640 liquid medium was used to dilute the inoculum suspension to a concentration of 5×10 4 CFU / ml with a final concentration of 2 times.

[0031] Operation steps: A 96-well plate was used, the first column was used as a blank control, the 12th column was used as a growth control, and the remaining columns were used for 2 times the final concentration of the two drugs alone and in combination. The drug amount of each strain was as shown in Table 1. The 1-8 wells of the 10th and 11th columns were used for 100 μl of single drug dilution, and the remaining wells were used for 50 μl of two-drug combination dilution. The concentration range of itraconazole was 0.0625-8 μg / ml, and the concentration range of eucalyptol was 62.5-8000 μg / ml. Each well was inoculated with 100 μl of the prepared bacterial suspension, and the drug sensitivity plate was placed in a 28°C constant temperature incubator for 5 days. The results of each strain in the experiment could be increased or decreased by 2 orders of magnitude in drug concentration.

[0032] 2. Experimental results

[0033] Table 1. Results of antibacterial experiments

[0034]

[0035] From Table 1, it can be seen that the results of the antibacterial experiments of itraconazole and eucalyptol show that itraconazole is more sensitive to the 15 strains of Microsporum canis clinically isolated, and the minimum inhibitory concentration ranges from 0.25 to 4 μg / ml; eucalyptol has a certain inhibitory effect on the 15 strains of Microsporum canis, and the minimum inhibitory concentration ranges from 250 to 4000 μg / ml; using the microdilution method, the minimum inhibitory concentration of itraconazole and eucalyptol combined for the 15 strains of Microsporum canis is 0.0625-0.5 μg / ml and 62.5-1000 μg / ml, according to the FICI calculation formula and determination standard, the FICI of all strains is ≤0.5, and itraconazole and eucalyptol show different degrees of synergistic effect on the 15 strains of Microsporum canis clinically isolated.

[0036] Example 2: Preparation of an external spray containing itraconazole and eucalyptol

[0037] The specific steps are as follows:

[0038] The external spray includes the following raw materials: itraconazole 9 g, eucalyptol 15 g, hydroxypropyl-β-cyclodextrin 250 g, propylene glycol 100 g, hydrochloric acid 9.5 g, diethylene glycol monoethyl ether 140 g, caprylic capric acid polyethylene glycol glycerol ester 40 g, sodium hydroxide 1.9 g, and the balance is purified water; wherein the amount of purified water is 1000 ml according to the preparation solution.

[0039] 1) Take the prescription amount of 45% purified water, heat to 80℃, add the prescription amount of propylene glycol, stir at 80℃ and 50 r / min for 3 min, to obtain solution 1;

[0040] 2) Slowly add the prescription amount of hydroxypropyl-β-cyclodextrin to solution 1, stir until completely dissolved, to obtain clear and transparent solution 2;

[0041] 3) Add the prescription amount of hydrochloric acid to solution 2, stir for 3 min, then add the prescription amount of itraconazole, stir for 45-60 min, to obtain clear solution 3;

[0042] 4) First, take the prescription amount of diethylene glycol monoethyl ether and caprylic capric acid polyethylene glycol glycerol ester, stir at 50 r / min for 3 min, then add the prescription amount of eucalyptol, stir for 10 min, to obtain light brown solution 4;

[0043] 5) Slowly add the obtained light brown solution 4 to the clear solution 3, stir for 20 min, then add the prescription amount of sodium hydroxide, stir until completely dissolved, finally add the balance of purified water to constant volume, stir for 30 min, pass through a 0.22 μm microporous filter, and then fill into a pressure container, to obtain the external spray containing itraconazole and eucalyptol.

[0044] Example 3: Preparation of an external spray containing itraconazole and eucalyptol

[0045] The specific steps are as follows:

[0046] The external spray includes the following raw materials: itraconazole 9 g, eucalyptol 15 g, hydroxypropyl-β-cyclodextrin 300 g, propylene glycol 140 g, hydrochloric acid 12 g, diethylene glycol monoethyl ether 160 g, caprylic capric acid polyethylene glycol glycerin ester 60 g, sodium hydroxide 2.1 g, and the balance is purified water; wherein the amount of purified water is 1000 ml.

[0047] 1) Take a batch of 45% purified water, heat to 80°C, add the prescribed amount of propylene glycol, 80°C, 50 r / min stirring for 3 min, get solution 1;

[0048] 2) Slowly add the prescribed amount of hydroxypropyl-β-cyclodextrin to solution 1, stir until completely dissolved, get clear and transparent solution 2;

[0049] 3) Add the prescribed amount of hydrochloric acid to solution 2, stir for 3 min, then add the prescribed amount of itraconazole, stir for 45-60 min, get clear solution 3;

[0050] 4) First take the prescribed amount of diethylene glycol monoethyl ether and caprylic capric acid polyethylene glycol glycerin ester, 50 r / min stirring for 3 min, then add the prescribed amount of eucalyptol, stir for 10 min, get light brown solution 4;

[0051] 5) Slowly add the light brown solution 4 obtained to the clear solution 3, stir for 20 min, then add the prescribed amount of sodium hydroxide, stir until completely dissolved, finally add the balance of purified water to constant volume, stir for 30 min, and pass through a 0.22 μm microporous filter, then fill into a pressure container to obtain an external spray containing itraconazole and eucalyptol.

[0052] Example 4: Preparation of an external spray containing itraconazole and eucalyptol

[0053] The specific steps are as follows:

[0054] The external spray includes the following raw materials: itraconazole 9 g, eucalyptol 15 g, hydroxypropyl-β-cyclodextrin 270 g, propylene glycol 120 g, hydrochloric acid 10.5 g, diethylene glycol monoethyl ether 150 g, caprylic capric acid polyethylene glycol glycerin ester 50 g, sodium hydroxide 2 g, and the balance is purified water; wherein the amount of purified water is 1000 ml.

[0055] 1) Take a batch of 45% purified water, heat to 80°C, add the prescribed amount of propylene glycol, 80°C, 50 r / min stirring for 3 min, get solution 1;

[0056] 2) Slowly add the prescribed amount of hydroxypropyl-β-cyclodextrin to solution 1, stir until completely dissolved, to obtain a clear and transparent solution 2;

[0057] 3) Add the prescribed amount of hydrochloric acid to solution 2, stir for 3 min, then add the prescribed amount of itraconazole, stir for 45-60 min, to obtain a clear solution 3;

[0058] 4) First take the prescribed amount of diethylene glycol monoethyl ether and caprylocaproyl macrogol glycerides, stir at 50 r / min for 3 min, then add the prescribed amount of eucalyptol, stir for 10 min, to obtain a light brown solution 4;

[0059] 5) Slowly add the obtained light brown solution 4 to the clear solution 3, stir for 20 min, then add the prescribed amount of sodium hydroxide, stir until completely dissolved, finally add the remaining purified water to constant volume, stir for 30 min, and pass through a 0.22 μm microporous filter, then fill into a pressure container to obtain an external use spray containing itraconazole and eucalyptol.

[0060] Example 5: Preparation of an external use spray containing itraconazole and eucalyptol

[0061] The specific steps are as follows:

[0062] The external use spray is composed of the following raw materials: itraconazole 9 g, eucalyptol 20 g, hydroxypropyl-β-cyclodextrin 270 g, propylene glycol 120 g, hydrochloric acid 10.5 g, diethylene glycol monoethyl ether 150 g, caprylocaproyl macrogol glycerides 50 g, sodium hydroxide 2 g, and the rest is purified water; wherein the amount of purified water is 1000 ml for preparation of the solution.

[0063] 1) Take 45% of the batch of purified purified water, heat to 80°C, add the prescribed amount of propylene glycol, stir at 50 r / min for 3 min at 80°C, to obtain solution 1;

[0064] 2) Slowly add the prescribed amount of hydroxypropyl-β-cyclodextrin to solution 1, stir until completely dissolved, to obtain a clear and transparent solution 2;

[0065] 3) Add the prescribed amount of hydrochloric acid to solution 2, stir for 3 min, then add the prescribed amount of itraconazole, stir for 45-60 min, to obtain a clear solution 3;

[0066] 4) First take the prescribed amount of diethylene glycol monoethyl ether and caprylocaproyl macrogol glycerides, stir at 50 r / min for 3 min, then add the prescribed amount of eucalyptol, stir for 10 min, to obtain a light brown solution 4;

[0067] 5) The light brown solution 4 obtained is slowly added to the clear solution 3, stirred for 20 min, then the prescribed amount of sodium hydroxide is added, stirred until completely dissolved, finally the remaining purified water is added to constant volume, stirred for 30 min, and after passing through a 0.22 μm microporous filter, it is filled into a pressure container to obtain an external use spray containing itraconazole and eucalyptol.

[0068] Example 6: Preparation of an external use spray containing itraconazole and eucalyptol

[0069] The specific steps are as follows:

[0070] The external use spray is composed of the following raw materials: itraconazole 10 g, eucalyptol 15 g, hydroxypropyl-β-cyclodextrin 270 g, propylene glycol 120 g, hydrochloric acid 10.5 g, diethylene glycol monoethyl ether 150 g, caprylic capric acid polyethylene glycol glycerol ester 50 g, sodium hydroxide 2 g, and the remaining purified water; wherein the amount of purified water is 1000 ml for the preparation of the solution.

[0071] 1) Take 45% of the batch of purified water, heat to 80°C, add the prescribed amount of propylene glycol, stir at 80°C and 50 r / min for 3 min to obtain solution 1;

[0072] 2) Slowly add the prescribed amount of hydroxypropyl-β-cyclodextrin to solution 1, stir until completely dissolved to obtain clear and transparent solution 2;

[0073] 3) Add the prescribed amount of hydrochloric acid solution to solution 2, stir for 3 min, then add the prescribed amount of itraconazole, stir for 45-60 min to obtain clear solution 3;

[0074] 4) First, take the prescribed amount of diethylene glycol monoethyl ether and caprylic capric acid polyethylene glycol glycerol ester, stir at 50 r / min for 3 min, then add the prescribed amount of eucalyptol, stir for 10 min to obtain light brown solution 4;

[0075] 5) Slowly add the light brown solution 4 obtained to the clear solution 3, stir for 20 min, then add the prescribed amount of sodium hydroxide, stir until completely dissolved, finally add the remaining purified water to constant volume, stir for 30 min, and after passing through a 0.22 μm microporous filter, it is filled into a pressure container to obtain an external use spray containing itraconazole and eucalyptol.

[0076] Example 7: Preparation of an external use spray containing itraconazole and eucalyptol

[0077] The specific steps are as follows:

[0078] The external use spray includes the following raw material composition: itraconazole 18g, eucalyptol 30g, hydroxypropyl-β-cyclodextrin 540g, propylene glycol 240g, hydrochloric acid 21g, diethylene glycol monoethyl ether 300g, caprylic capric acid polyethylene glycol glycerin ester 100g, sodium hydroxide 4g, and the balance is purified water; wherein the amount of purified water is 2000ml according to the preparation solution.

[0079] 1) Take 45% of the batch of purified water, heat to 80°C, add the prescription amount of propylene glycol, 80°C, 50r / min stirring for 3min, get solution 1;

[0080] 2) Slowly add the prescription amount of hydroxypropyl-β-cyclodextrin to solution 1, stir until completely dissolved, get clear transparent solution 2;

[0081] 3) Add the prescription amount of hydrochloric acid to solution 2, stir for 3min, then add the prescription amount of itraconazole, stir for 45~60min, get clear solution 3;

[0082] 4) First take the prescription amount of diethylene glycol monoethyl ether and caprylic capric acid polyethylene glycol glycerin ester, 50r / min stirring for 3min, then add the prescription amount of eucalyptol, stir for 10min, get light brown solution 4;

[0083] 5) Slowly add the light brown solution 4 obtained to the clear solution 3, stir for 20min, then add the prescription amount of sodium hydroxide, stir until completely dissolved, finally add the balance of purified water to constant volume, stir for 30min, and pass through 0.22μm microporous filter, then fill into a pressure container, to obtain the external use spray containing itraconazole and eucalyptol.

[0084] Comparative example 1: preparation of an external use spray containing itraconazole and eucalyptol

[0085] The specific steps are as follows:

[0086] The external use spray includes the following raw material composition: itraconazole 18g, eucalyptol 30g, hydroxypropyl-β-cyclodextrin 540g, propylene glycol 240g, hydrochloric acid 21g, diethylene glycol monoethyl ether 300g, caprylic capric acid polyethylene glycol glycerin ester 100g, sodium hydroxide 4g, and the balance is purified water; wherein the amount of purified water is 2000ml according to the preparation solution.

[0087] 1) Take 45% of the batch of purified water, heat to 80°C, add the prescription amount of propylene glycol, 80°C, 50r / min stirring for 3min, get solution 1;

[0088] 2) Slowly add the prescription amount of hydroxypropyl-β-cyclodextrin to solution 1, stir until completely dissolved, get clear transparent solution 2;

[0089] 3) Add the prescribed amount of hydrochloric acid to solution 2, stir for 3 minutes, then add the prescribed amount of itraconazole, stir for 45-60 minutes, to obtain clear solution 3;

[0090] 4) First take the prescribed amount of diethylene glycol monoethyl ether and caprylocaproyl macrogolglycerides, stir at 50 r / min for 3 minutes, then add the prescribed amount of eucalyptol, stir for 10 minutes, to obtain light brown solution 4;

[0091] 5) Slowly add the obtained light brown solution 4 to clear solution 3, stir for 20 minutes, then add the prescribed amount of sodium hydroxide, stir until completely dissolved, finally add the remaining purified water to constant volume, stir for 30 minutes, and after passing through a 0.22 μm microporous filter, fill into a pressure container, to obtain the external use spray containing itraconazole and eucalyptol.

[0092] Comparative Example 2: Preparation of an external use spray containing itraconazole and eucalyptol

[0093] The specific steps are as follows:

[0094] The external use spray comprises the following raw materials: itraconazole 9 g, eucalyptol 15 g, hydroxypropyl-β-cyclodextrin 270 g, propylene glycol 120 g, hydrochloric acid 10.5 g, diethylene glycol monoethyl ether 120 g, caprylocaproyl macrogolglycerides 30 g, sodium hydroxide 2 g, and the remaining amount is purified water; wherein the amount of purified water is 1000 ml based on the preparation of the solution.

[0095] 1) Take 45% of the batch of purified water, heat to 80°C, add the prescribed amount of propylene glycol, stir at 50 r / min for 3 minutes at 80°C, to obtain solution 1;

[0096] 2) Slowly add the prescribed amount of hydroxypropyl-β-cyclodextrin to solution 1, stir until completely dissolved, to obtain clear and transparent solution 2;

[0097] 3) Add the prescribed amount of hydrochloric acid to solution 2, stir for 3 minutes, then add the prescribed amount of itraconazole, stir for 45-60 minutes, to obtain clear solution 3;

[0098] 4) First take the prescribed amount of diethylene glycol monoethyl ether and caprylocaproyl macrogolglycerides, stir at 50 r / min for 3 minutes, then add the prescribed amount of eucalyptol, stir for 10 minutes, to obtain light brown solution 4;

[0099] 5) Slowly add the obtained light brown solution 4 to clear solution 3, stir for 20 minutes, then add the prescribed amount of sodium hydroxide, stir until completely dissolved, finally add the remaining purified water to constant volume, stir for 30 minutes, and after passing through a 0.22 μm microporous filter, fill into a pressure container, to obtain the external use spray containing itraconazole and eucalyptol.

[0100] Preparation of a topical spray containing only itraconazole

[0101] The specific steps are as follows:

[0102] The topical spray includes the following raw materials: itraconazole 9 g, hydroxypropyl-β-cyclodextrin 270 g, propylene glycol 120 g, hydrochloric acid 10.5 g, diethylene glycol monoethyl ether 150 g, caprylocaproyl macrogol glycerides 50 g, sodium hydroxide 2 g, and the balance is purified water; wherein the amount of purified water is determined by preparing a solution to a volume of 1000 ml.

[0103] 1) Take a batch of 45% purified water, heat to 80°C, add the prescribed amount of propylene glycol, 80°C, 50 r / min stirring for 3 min, get solution 1;

[0104] 2) Slowly add the prescribed amount of hydroxypropyl-β-cyclodextrin to solution 1, stir until completely dissolved, get clear and transparent solution 2;

[0105] 3) Add the prescribed amount of hydrochloric acid to solution 2, stir for 3 min, then add the prescribed amount of itraconazole, stir for 45-60 min, get clear solution 3;

[0106] 4) First take the prescribed amount of diethylene glycol monoethyl ether and caprylocaproyl macrogol glycerides, 50 r / min stirring for 3 min, get light brown solution 4;

[0107] 5) Slowly add the light brown solution 4 obtained to the clear solution 3, stir for 20 min, then add the prescribed amount of sodium hydroxide, stir until completely dissolved, finally add the balance of purified water to volume, stir for 30 min, and pass through a 0.22 μm microporous filter, then fill into a pressure container to obtain a topical spray containing only itraconazole.

[0108] Preparation of a topical spray containing only eucalyptol

[0109] The specific steps are as follows:

[0110] The topical spray includes the following raw materials: eucalyptol 15 g, hydroxypropyl-β-cyclodextrin 270 g, propylene glycol 120 g, hydrochloric acid 10.5 g, diethylene glycol monoethyl ether 150 g, caprylocaproyl macrogol glycerides 50 g, sodium hydroxide 2 g, and the balance is purified water; wherein the amount of purified water is determined by preparing a solution to a volume of 1000 ml.

[0111] 1) Take a batch of 45% purified water, heat to 80°C, add the prescribed amount of propylene glycol, 80°C, 50 r / min stirring for 3 min, get solution 1;

[0112] 2) Slowly add the prescribed amount of hydroxypropyl-β-cyclodextrin in solution 1, stir until completely dissolved, to obtain a clear transparent solution 2;

[0113] 3) Add the prescribed amount of hydrochloric acid in solution 2, stir for 3 min, to obtain a clear solution 3;

[0114] 4) First, take the prescribed amount of diethylene glycol monoethyl ether and caprylocaproyl polyoxylglycerides, stir at 50 r / min for 3 min, then add the prescribed amount of eucalyptol, stir for 10 min, to obtain a light brown solution 4;

[0115] 5) Slowly add the obtained light brown solution 4 to the clear solution 3, stir for 20 min, then add the prescribed amount of sodium hydroxide, stir until completely dissolved, finally add the remaining purified water to constant volume, stir for 30 min, and pass through a 0.22 μm microporous filter, then fill into a pressure container, to obtain an external use spray containing only eucalyptol.

[0116] Example 8: Stability experiment

[0117] 1. Experimental materials

[0118] The external use sprays prepared in Comparative Examples 1-2 and Examples 2-7

[0119] 2. Experimental method

[0120] The eight batches of external use sprays containing itraconazole and eucalyptol in Comparative Examples 1-2 and Examples 2-7 were subjected to accelerated stability test. The specific method of accelerated stability test was as follows: different batches of products were placed in a pharmaceutical accelerated stability chamber, the temperature was set to 40℃ and the relative humidity was set to 75%, samples were taken at 0, 1, 2, 3 and 6 months, and the product properties, pH value, relative density, and itraconazole and eucalyptol contents were detected.

[0121] 3. Experimental results

[0122] Table 2: Accelerated stability test results

[0123]

[0124] As shown in Table 2, the accelerated stability test results of the product showed that the product obtained in Comparative Example 1 failed to pass the accelerated stability test, its properties, relative density and itraconazole content all changed significantly, which did not meet the product quality requirements, indicating that reducing the amount of solubilizing agent hydroxypropyl-β-cyclodextrin and cosolvent propylene glycol in the preparation prescription could significantly affect the product quality;

[0125] The products obtained in Examples 2-7 are subjected to accelerated stability test, and the properties, pH value, relative density, itraconazole and eucalyptol content involved in the product quality requirements have no significant change. The accelerated stability test results of the products obtained in Examples 4 and 7 show that the product quality has no significant difference in the production batch amplification process by the preparation method. Therefore, the external spray containing itraconazole and eucalyptol produced by the present application has good process reproducibility and stable product quality.

[0126] Example 9: In vitro transdermal penetration experiment

[0127] 1. Experimental materials

[0128] The external sprays prepared in Comparative Example 2, Example 4 and Example 7

[0129] 2. Experimental method

[0130] The in vitro transdermal penetration test is carried out on the three batches of external sprays containing itraconazole and eucalyptol in Comparative Example 2, Example 4 and Example 7. The treated rat skin is fixed on the receiving port of Franz diffusion cell, 0.9% sodium chloride solution is used as the receiving liquid to simulate the physiological environment of the skin, and the device is placed on the intelligent drug transdermal diffusion instrument with a temperature of (32±0.5)℃. The samples are taken at 0.25, 0.5, 1, 2, 4, 6, 8 and 12 hours, respectively, and the amounts of itraconazole and eucalyptol are measured to calculate the cumulative permeation amount (Jss) of the drug, so as to evaluate the in vitro transdermal performance and characteristics of the product. Q

[0131] 3. Experimental results

[0132] Table 3: In vitro transdermal penetration cumulative drug release amount (Jss) Q

[0133]

[0134] From Table 3, Figures 1-2 ​​It can be seen that the in-vitro transdermal penetration test results show that the cumulative permeation amounts of the three groups of tested products increase with the extension of time, but the cumulative permeation amount of the product obtained in Comparative Example 2 is the smallest, indicating that the transdermal penetration performance of the product is the lowest, and the transdermal penetration performances of the products obtained in Examples 4 and 7 have no obvious difference; it can be known from the transdermal penetration performance results of the product obtained in Comparative Example 2 that reducing the amounts of the skin penetration promoters diethylene glycol monoethyl ether and caprylic capric acid polyethylene glycol glycerin ester in the preparation prescription can obviously affect the transdermal penetration performance of the product. At the same time, the transdermal penetration test results of the products obtained in Examples 4 and 7 also show that the transdermal penetration performances of the products obtained in the production batch amplification process of the preparation method have no obvious difference. Therefore, the external spray containing itraconazole and eucalyptol prepared by the present application has excellent transdermal penetration performance, can be used through the external spray mode and the local transdermal route, and can achieve the therapeutic effect of treating superficial skin fungal infections.

[0135] Example 10: Clinical treatment effect verification

[0136] 1. Experimental materials

[0137] the external spray obtained in Example 4,

[0138] 2. Experimental method

[0139] Two cats with fungal skin diseases identified positive for Microsporum canis were selected as Case 1 and Case 2, and the lesion sites showed symptoms such as hair loss, skin redness, and itching. The product obtained in Example 4 was used, and the lesion sites were treated with a dose of 2 sprays per 4 cm 2 The skin of the affected area was sprayed with a dose of 2 sprays per 4 cm

[0140] 3. Experimental results

[0141] As shown in Figures 3-4 , the clinical treatment effect verification results show that the two cats with confirmed diagnosis no longer show itching after 3 weeks of continuous administration, the continuous hair loss is significantly improved, the skin returns to normal, and after a subsequent 3-week recovery period, the hair on the lesion site regrows and returns to normal, and no recurrence occurs. The clinical treatment effect of the product prepared by the present application is significant.

[0142] Example 11: Clinical treatment effect comparison experiment

[0143] 1. Experimental materials

[0144] the external sprays obtained in Examples 4, Comparative Example 3, and Comparative Example 4

[0145] 2. Experimental method

[0146] Forty-five cats with fungal skin diseases identified positive for Microsporum canis were selected and randomly divided into three groups, and each group was treated with a dose of 2 sprays per 4 cm 2The dose of the product obtained in Example 4, the product of Comparative Example 3 and the product of Comparative Example 4 was sprayed on the skin of the affected area twice a day for 3 weeks.

[0147] 3. Experimental results

[0148] Table 4. Comparison of clinical treatment effect experimental results

[0149]

[0150] As shown in Table 4, the cure rates of the cats in the Example 4 group, the Comparative Example 3 group and the Comparative Example 4 group were 86.7%, 26.7% and 13.3% respectively, and the total effective rates were 100.0%, 73.4% and 40.0% respectively. The cure rate and the total effective rate of the product obtained in Example 4 were both significantly higher than those of the two comparative examples, and the clinical treatment effect was the best, indicating that the external spray prepared by combining itraconazole and eucalyptol had a significant clinical effect and better treatment effect, and had great advantages.

[0151] Finally, it should be pointed out that the above only describes the preferred embodiments of the present application and is not intended to limit the present application. Although the present application has been described in detail with reference to the foregoing embodiments, those skilled in the art can still modify the technical solutions described in the foregoing embodiments or replace some of the technical features with equivalent ones. Any modification, equivalent replacement, improvement, etc. made within the spirit and principles of the present application shall be included in the protection scope of the present application.

Claims

1. An external use spray containing itraconazole and eucalyptol, characterized in that, The material composition in each 1000ml external use spray: itraconazole 8~10g, eucalyptol 10~20g, hydroxypropyl-β-cyclodextrin 250~300g, propylene glycol 100~140g, hydrochloric acid 9.5~12g, diethylene glycol monoethyl ether 140~160g, caprylocaproyl macrogol glycerides 40~60g, sodium hydroxide 1.9~2.1g, and the rest is purified water; wherein the amount of purified water is determined by preparing solution to 1000ml; The preparation method of the external use spray, the specific steps are as follows: 1) take the prescription amount of 45% purified water, heat to 80℃, add the prescription amount of propylene glycol, 80℃, 50r / min stirring 3min, get solution 1; 2) slowly add the prescription amount of hydroxypropyl-β-cyclodextrin in solution 1, stir until completely dissolved, get clear transparent solution 2; 3) add the prescription amount of hydrochloric acid in solution 2, stir 3min, then add the prescription amount of itraconazole, stir 45~60min, get clear solution 3; 4) first take the prescription amount of diethylene glycol monoethyl ether and caprylocaproyl macrogol glycerides, 50r / min stirring 3min, then add the prescription amount of eucalyptol, stir 10min, get light brown solution 4; 5) slowly add the obtained light brown solution 4 to the clear solution 3, stir 20min, then add the prescription amount of sodium hydroxide, stir until completely dissolved, finally add the rest of the purified water to constant volume, stir 30min, and pass through 0.22μm microporous filter, then fill into pressure container, get the external use spray containing itraconazole and eucalyptol.

2. The external spray containing itraconazole and eucalyptol according to claim 1, characterized in that, The material composition in each 1000ml external use spray: Itraconazole 9g, eucalyptol 15g, hydroxypropyl-β-cyclodextrin 270g, propylene glycol 120g, hydrochloric acid 10.5g, diethylene glycol monoethyl ether 150g, caprylocaproyl macrogol glycerides 50g, sodium hydroxide 2g, and the rest is purified water; wherein the amount of purified water is determined by preparing solution to 1000ml.

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