Brutenafine inclusion compound hydrogel for eyes and preparation method of butenafine inclusion compound hydrogel

Abstract

The invention relates to an ophthalmic butenafine clathrate compound hydrogel, the hydrogel comprises a butenafine clathrate compound, butenafine is clathrated through a solubilizer hydroxypropyl-beta-cyclodextrin or sulfobutyl-beta-cyclodextrin, the hydrophobicity of a medicine is improved, the solubility of the medicine is improved, the bioavailability of the medicine is improved, and the bioavailability of the medicine is improved. When acting on eyes together with the temperature-sensitive gel matrix, the medicine does not generate irritation and foreign body sensation, so that the medicine utilization rate is increased; the hydrogel further comprises a temperature-sensitive gel matrix, and the temperature-sensitive gel matrix is prepared from poloxamer, hydroxypropyl methyl cellulose and sodium hyaluronate; the poloxamer is a temperature-sensitive material, so that the medicine is changed into soft gel after being applied to eyes; the hydroxypropyl methyl cellulose can enhance the mechanical strength of the gel, improve the cornea adhesive force of the gel and prolong the residence time of eyes; the sodium hyaluronate is high in water retention capacity; animal experiments prove that the eye retention time of the butenafine inclusion compound hydrogel prepared by the method disclosed by the invention is at least 120 minutes, which is far higher than that of a butenafine suspension, so that the medication frequency is greatly reduced.

Description

Technical Field

[0001] The invention belongs to the field of medicine preparation, and particularly relates to butenafine inclusion compound hydrogel for eyes and a preparation method thereof. Background Art

[0002] At present, about 90% of clinical ophthalmic topical medications are traditional eye drops or eye ointments, which have the advantages of avoiding systemic administration, being easy to use, and concentrating the drug on the affected area. However, the special physiological barriers and clearance mechanisms of the eye are the biggest barriers to topical ocular medications. Physiological barriers such as the cornea, conjunctiva, tear drainage, and reflexes (blinking and tearing) greatly limit the effects of drugs, resulting in the bioavailability of conventional dosage forms of drugs in the eye being generally less than 10% (about 5%). Butenafine (BTF) is a new type of allylamine antifungal drug and a rising star in antifungal drugs. Its mechanism of action is to selectively inhibit fungal squalene epoxidase and prevent fungal ergosterol synthesis. In addition, butenafine is poorly soluble in water and also has the problem of low bioavailability. Summary of the invention

[0003] In view of the shortcomings of the prior art, the present invention provides an ophthalmic butenafine inclusion compound hydrogel and a preparation method thereof.

[0004] The technical solution of the present invention is:

[0005] The butenafine inclusion compound hydrogel for eye use comprises butenafine inclusion compound and a thermosensitive gel matrix. The mass ratio of the butenafine inclusion compound to the thermosensitive gel matrix is ​​(1-1.5):(10-15), and the mass concentration of the thermosensitive gel matrix is ​​17-28.1%.

[0006] Furthermore, the butenafine inclusion compound comprises butenafine, anhydrous ethanol, a solubilizing agent, and a lyophilizing protective agent, the molar ratio of butenafine to the solubilizing agent is 1:2, the mass ratio of butenafine:lyophilizing protective agent is (1-5):(10-20), and the mass concentration of butenafine after being dissolved in anhydrous ethanol solution is 40-60%.

[0007] Furthermore, the solubilizing agent is at least one of hydroxypropyl-β-cyclodextrin or sulfobutyl-β-cyclodextrin.

[0008] Furthermore, the lyoprotectant is at least one of glycine or mannitol.

[0009] Furthermore, the osmotic pressure regulator is sodium chloride.

[0010] Furthermore, the thermosensitive gel matrix includes poloxamer, hydroxypropyl methylcellulose, sodium hyaluronate, and an osmotic pressure regulator, and the mass ratio of poloxamer:hydroxypropyl methylcellulose:sodium hyaluronate:osmotic pressure regulator is (15-25):(1-2):(0.1-0.2):0.9.

[0011] The preparation method of the butenafine inclusion compound hydrogel for eyes comprises the following steps:

[0012] S1: dissolving butenafine in anhydrous ethanol to obtain a butenafine ethanol solution, adding the butenafine ethanol solution to a solubilizing agent, mixing, filtering, adding a lyophilization protective agent to the filtrate, and freeze-drying to obtain a butenafine inclusion compound;

[0013] S2: Dissolve hydroxypropyl methylcellulose and sodium hyaluronate in the prescribed proportion, add poloxamer in the prescribed proportion and stir, let stand for 18-24 hours until transparent, then add an osmotic pressure regulator, and finally add the butenafine inclusion compound in the prescribed proportion prepared in step S1 and stir to dissolve to obtain an ophthalmic butenafine inclusion compound hydrogel.

[0014] Furthermore, the filtration in step S1 is performed by filtering using a 0.3-0.5 μm microporous filter membrane.

[0015] Compared with the prior art, the present invention has at least the following advantages:

[0016] 1. The present invention relates to an ophthalmic butenafine inclusion compound hydrogel, wherein the hydrogel comprises a butenafine inclusion compound. The present invention includes butenafine by encapsulating butenafine with a solubilizer (hydroxypropyl-β-cyclodextrin or sulfobutyl-β-cyclodextrin), thereby improving the hydrophobicity of the drug and greatly increasing the solubility of the drug component. When the butenafine is applied to the eye together with a thermosensitive gel matrix, it does not cause irritation or foreign body sensation, thereby improving the utilization rate of the drug.

[0017] 2. The present invention relates to an ophthalmic butenafine inclusion hydrogel, wherein the hydrogel further comprises a thermosensitive gel matrix, which is composed of poloxamer, hydroxypropylmethylcellulose and sodium hyaluronate; poloxamer is a thermosensitive material, which allows the drug to change into a soft gel after being used in the eye; hydroxypropylmethylcellulose can enhance the mechanical strength of the gel, improve the corneal adhesion of the gel, and increase the eye retention time of the hydrogel drug; sodium hyaluronate has a strong water retention performance, which allows the eyes to remain moisturized for a long time; experimental verification shows that the eye retention time of the butenafine inclusion hydrogel prepared by the method of the present invention is at least 120 minutes, which is much higher than that of the butenafine suspension, greatly reducing the number of medication times; and the antibacterial effect of the butenafine inclusion hydrogel prepared by the method of the present invention is higher than that of the butenafine suspension and amphotericin B eye drops. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] In order to more clearly illustrate the specific embodiments of the present invention, the drawings required for use in the specific embodiments or the description of the prior art will be briefly introduced below.

[0019] Figure 1 This is the appearance of the BTF inclusion compound prepared in Example 2 of the present invention. DETAILED DESCRIPTION

[0020] The present invention is further described in detail below. It is necessary to point out that the following specific implementation methods are only used to further illustrate the present invention and cannot be understood as limiting the scope of protection of the present invention. Technicians in this field can make some non-essential improvements and adjustments to the present invention based on the above application content.

[0021] The present invention provides a general and / or specific description of the materials and test methods used in the experiment. The test methods or test methods involved are all conventional methods unless otherwise specified; the reagents or instruments used are all conventional products available on the market, prepared or used in a conventional manner, unless the manufacturer is specified.

[0022] Sources:

[0023] Hydroxypropyl methylcellulose: average molecular weight 10000, purchased from Sigma;

[0024] Amphotericin B eye drops: North China Pharmaceutical.

[0025] Example 1 Example 1 is a method for preparing butenafine inclusion compound hydrogel

[0026] S1: 0.1 g of butenafine was dissolved in 200 μL of anhydrous ethanol to prepare a BTF anhydrous ethanol solution with a mass concentration of 50%, the molar ratio of BTF to sulfobutyl-β-cyclodextrin inclusion material was 1:2, the sulfobutyl-β-cyclodextrin inclusion material was dissolved in 5 mL of sterile water for injection, the BTF anhydrous ethanol solution was slowly dripped into the sulfobutyl-β-cyclodextrin aqueous solution at a temperature of 45°C and a stirring speed of 400 r / min, the stirring was continued for 3 hours, and the solution was allowed to stand for 3 hours, and the solution was filtered through a 0.45 μm microporous filter membrane to obtain a filtrate, 2 g of glycine was added to the filtrate as a lyophilization protective agent, and then cooled The freeze-drying procedure is as follows: C1: -40°C, 120 min; C2: -20°C, 80 min; C3: -20°C, 60 min; C4: -15°C, 60 min; C5: -15°C, 600 min; C6: -10°C, 90 min; C7: -10°C, 60 min; C8: 0°C, 180 min; C9: 0°C, 90 min; C10: 20°C, 480 min; C11: 20°C, 60 min. The freeze-dried inclusion compound is obtained after the procedure is completed. The freeze-dried inclusion compound is ground and passed through an 80-mesh sieve for standby use to obtain the BTF inclusion compound. Three batches are prepared for standby use.

[0027] S2: Take 0.1g of hydroxypropyl methylcellulose and 0.01g of sodium hyaluronate and dissolve them completely in 10mL of sterile water for injection, then add 2g of gel matrix poloxamer, place the prepared solution in a 4°C refrigerator for 24h, wait until the solution becomes transparent, add 0.09g of sodium chloride as an osmotic pressure regulator, and finally add 1g of the BTF inclusion complex prepared in step S1, and dissolve under gentle stirring to obtain butenafine inclusion complex hydrogel.

[0028] The BTF inclusion compound prepared in Example S1 of this embodiment is in the form of loose white powder, which is a colorless transparent solution after being dissolved in sterile water for injection.

[0029] Example 2 This example is a method for preparing butenafine inclusion compound hydrogel

[0030] S1: 0.3 g of butenafine was dissolved in 600 μL of anhydrous ethanol to prepare a BTF anhydrous ethanol solution with a mass concentration of 50%, the molar ratio of BTF to sulfobutyl-β-cyclodextrin inclusion material was 1:2, the sulfobutyl-β-cyclodextrin inclusion material was dissolved in 8 mL of sterile water for injection, the BTF anhydrous ethanol solution was slowly dripped into the sulfobutyl-β-cyclodextrin aqueous solution at a temperature of 45°C and a stirring speed of 400 r / min, the stirring was continued for 3 hours, the solution was allowed to stand for 3 hours, the solution was filtered through a 0.45 μm microporous filter membrane to obtain a filtrate, 1 g of glycine was added to the filtrate as a lyophilization protective agent, and then cooled The freeze-drying procedure is as follows: C1: -40°C, 120 min; C2: -20°C, 80 min; C3: -20°C, 60 min; C4: -15°C, 60 min; C5: -15°C, 600 min; C6: -10°C, 90 min; C7: -10°C, 60 min; C8: 0°C, 180 min; C9: 0°C, 90 min; C10: 20°C, 480 min; C11: 20°C, 60 min. The freeze-dried inclusion compound is obtained after the procedure is completed. The freeze-dried inclusion compound is ground and passed through an 80-mesh sieve for standby use to obtain the BTF inclusion compound. Three batches are prepared for standby use.

[0031] S2: Take 0.2g of hydroxypropyl methylcellulose and 0.02g of sodium hyaluronate and dissolve them in 10mL of sterile water for injection until they are completely dissolved. Then add 1.5g of gel matrix poloxamer thereto. Place the prepared solution in a refrigerator at 4°C for 24h. When the solution becomes transparent, add 0.09g of sodium chloride as an osmotic pressure regulator. Finally, add 1.5g of the BTF inclusion complex prepared in step S1 and dissolve under gentle stirring to obtain butenafine inclusion complex hydrogel.

[0032] The BTF inclusion compound prepared in Example S1 has the appearance of loose white powder ( Figure 1 ), which is a colorless and transparent solution after being dissolved in sterile water for injection.

[0033] Example 3 This example is a method for preparing butenafine inclusion compound hydrogel

[0034] S1: 0.5 g of butenafine was dissolved in 400 μL of anhydrous ethanol to prepare a BTF anhydrous ethanol solution with a mass concentration of 50%, the molar ratio of BTF to hydroxypropyl-β-cyclodextrin inclusion material was 1:2, the hydroxypropyl-β-cyclodextrin inclusion material was dissolved in 10 mL of sterile water for injection, the BTF anhydrous ethanol solution was slowly dripped into the hydroxypropyl-β-cyclodextrin aqueous solution at a temperature of 45°C and a stirring speed of 400 r / min, the stirring was continued for 3 hours, and the solution was allowed to stand for 3 hours, and the solution was filtered through a 0.45 μm microporous filter membrane to obtain a filtrate, 2 g of mannitol was added to the filtrate as a lyophilization protective agent, and then cooled. The freeze-drying procedure is as follows: C1: -40°C, 120 min; C2: -20°C, 80 min; C3: -20°C, 60 min; C4: -15°C, 60 min; C5: -15°C, 600 min; C6: -10°C, 90 min; C7: -10°C, 60 min; C8: 0°C, 180 min; C9: 0°C, 90 min; C10: 20°C, 480 min; C11: 20°C, 60 min. The freeze-dried inclusion compound is obtained after the procedure is completed. The freeze-dried inclusion compound is ground and passed through an 80-mesh sieve for standby use to obtain the BTF inclusion compound. Three batches are prepared for standby use.

[0035] S2: Take 0.1g of hydroxypropyl methylcellulose and 0.02g of sodium hyaluronate and dissolve them in 10mL of sterile water for injection until they are completely dissolved. Then add 2.3g of gel matrix poloxamer thereto. Place the prepared solution in a refrigerator at 4°C for 24h. When the solution becomes transparent, add 0.09g of sodium chloride as an osmotic pressure regulator. Finally, add 1.5g of the BTF inclusion complex prepared in step S1 and dissolve under gentle stirring to obtain butenafine inclusion complex hydrogel.

[0036] The BTF inclusion compound prepared in Example S1 of this embodiment is in the form of loose white powder, which is a colorless transparent solution after being dissolved in sterile water for injection.

[0037] Test Example 1 Determination of the solubility of butenafine (BTF) inclusion complex

[0038] The HPLC method for determining the content of BTF inclusion complex is as follows: chromatographic column: C 18 Column; mobile phase: acetonitrile-methanol-0.05mol / L ammonium acetate buffer salt = 65:20:15; detection wavelength: 282nm; injection volume: 10μL; flow rate: 1.0mL / min, column temperature: 30℃.

[0039] First, 800 mg of BTF inclusion complex was weighed to prepare a supersaturated solution, shaken at 25°C ± 0.5°C for 72 h, centrifuged at 4000 rpm for 5 min, the supernatant was taken, filtered through a 0.45 μm microporous filter membrane, and after dilution, the solubility was determined according to the above-mentioned HPLC method. The solubility calculation formula was as follows: solubility = concentration × dilution factor.

[0040] This test example uses the BTF inclusion compound prepared in Example 2 as an example. The solubility of the BTF inclusion compound in 30°C water is about 1.05 mg / ml, which is slightly soluble in water. Pure BTF is difficult to dissolve in water. The present invention greatly improves the solubility of BTF by solubilizing the inclusion compound.

[0041] Test Example 2 Determination of drug loading of BTF inclusion compound

[0042] This test example uses the BTF inclusion compound prepared in Example 2 as an example. 20 mg of the BTF inclusion compound is accurately weighed into a 10 mL volumetric flask, 5 mL of methanol is added, and ultrasonic oscillation is performed for 15 minutes. After the BTF is completely dissolved, the volume is diluted and fixed, and the drug loading amount is calculated according to the following formula: drug loading amount % = amount of BTF in the preparation / total weight of the preparation × 100%.

[0043] In this test example, the drug loading of the BTF inclusion compound tested by the above method was 10.3%.

[0044] Test Example 3 Evaluation of the antifungal effect of BTF inclusion complex hydrogel in vitro

[0045] Aspergillus is made into 10 6 CFU / ml bacterial suspension was inoculated into glucose agar medium, and the Oxford cup was placed into the above bacterial medium using a steel tube placement device. BTF suspension (BTF eye drops were not commercially available, so BTF suspension was prepared instead), BTF inclusion hydrogel (the BTF inclusion hydrogel prepared in Example 2 was used as an example in this test example), amphotericin B eye drops, and sterile water for injection were added to different Oxford cups, and then transferred to a biochemical incubator and cultured at 28°C for 72h. The size of the inhibition zone formed by the four solutions was measured with a vernier caliper. The results are shown in Table 1.

[0046] The concentration of BTF suspension is 0.3%; the concentration of BTF inclusion compound hydrogel is 0.3%; the concentration of amphotericin B eye drops is 0.3%.

[0047] Table 1 Inhibition zone size of different drugs against Aspergillus (n=3)

[0048] Group Inhibition zone measurement value (mm) BTF solution (suspension) 9.6±1.1 BTF inclusion complex hydrogel 18.3±1.6 Amphotericin B eye drops 15.7±1.4 Sterile water for injection 0

[0049] As can be seen from Table 1, the inhibition zone formed by the BTF inclusion compound hydrogel is the highest, which is significantly higher than that of the BTF suspension group and the eye drop group, indicating that the effect of the BTF inclusion compound hydrogel prepared in Example 2 is better than that of other drugs at the same dose.

[0050] Test Example 4 Investigation of BTF inclusion complex hydrogel eye retention time

[0051] Weigh 6 mg of sodium fluorescein, add it to 10 ml of BTF inclusion complex hydrogel, mix well and set aside; weigh 6 mg of sodium fluorescein, add it to BTF solution (suspension) and set aside.

[0052] Two rabbits were fixed in a rabbit box, the lower eyelids were lifted, the conjunctival sac was pulled into a ring, and the left eye was given a BTF solution (suspension) containing sodium fluorescein at a dose of 0.3%, and the right eye was given a BTF inclusion compound hydrogel containing sodium fluorescein at a dose of 0.3%. After administration, the eyes were closed for 10 seconds, and then images were collected under a slit lamp. The images were recorded every 5 minutes for the first 30 minutes, and every 15 minutes after 30 minutes. The retention time of the drug containing sodium fluorescein in the eye was observed until there was no fluorescence. The results are shown in Table 2, where "-" means no fluorescein, "+" means there is fluorescein but it is weak, and "++" means there is fluorescein and it is strong.

[0053] Table 2 Comparison of drug intraocular retention time

[0054] time BTF solution BTF inclusion complex hydrogel 0min(before administration) — — 5min ++ ++ 10min + ++ 15min + ++ 20min — ++ 25min — ++ 30min — ++ 45min — ++ 60min — ++ 75min — + 90min — + 105min — + 120min — +

[0055] As can be seen from Table 2, compared with BTF suspension, BTF inclusion complex hydrogel can stay in the eye for at least 120 min, effectively reducing the number of dosing times.

[0056] The above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or replace some or all of the technical features therein by equivalents. These modifications or replacements do not make the essence of the corresponding technical solutions deviate from the scope of the technical solutions of the embodiments of the present invention, and they should all be included in the scope of the claims and specification of the present invention.

Claims

1. An ophthalmic butenafine inclusion compound hydrogel, characterized in that: The butenafine inclusion compound hydrogel comprises butenafine inclusion compound and a thermosensitive gel matrix. The mass ratio of the butenafine inclusion compound to the thermosensitive gel matrix is ​​(1-1.5):(10-15), and the mass concentration of the thermosensitive gel matrix is ​​17-28.1%.

2. The ophthalmic butenafine inclusion compound hydrogel according to claim 1, characterized in that: The butenafine inclusion compound comprises butenafine, anhydrous ethanol, a solubilizing agent, and a lyophilizing protective agent. The molar ratio of the butenafine to the solubilizing agent is 1:2, the mass ratio of the butenafine to the lyophilizing protective agent is (1-5):(10-20), and the mass concentration of the butenafine after being dissolved in anhydrous ethanol solution is 40-60%.

3. The ophthalmic butenafine inclusion compound hydrogel according to claim 2, characterized in that: The solubilizing agent is at least one of hydroxypropyl-β-cyclodextrin or sulfobutyl-β-cyclodextrin.

4. The ophthalmic butenafine inclusion compound hydrogel according to claim 2, characterized in that: The lyophilization protectant is at least one of glycine or mannitol.

5. The ophthalmic butenafine inclusion compound hydrogel according to claim 2, characterized in that: The osmotic pressure regulator is sodium chloride.

6. The ophthalmic butenafine inclusion compound hydrogel according to claim 1, characterized in that: The thermosensitive gel matrix includes poloxamer, hydroxypropyl methylcellulose, sodium hyaluronate, and an osmotic pressure regulator, and the mass ratio of poloxamer:hydroxypropyl methylcellulose:sodium hyaluronate:osmotic pressure regulator is (15-25):(1-2):(0.1-0.2):0.

9.

7. The method for preparing the butenafine inclusion compound hydrogel for ophthalmology according to any one of claims 1 to 6, characterized in that: The following steps are involved: S1: dissolving butenafine in anhydrous ethanol to obtain a butenafine ethanol solution, adding the butenafine ethanol solution to a solubilizing agent, mixing, filtering, adding a lyophilization protective agent to the filtrate, and freeze-drying to obtain a butenafine inclusion compound; S2: Dissolve hydroxypropyl methylcellulose and sodium hyaluronate in the prescribed proportion, add poloxamer in the prescribed proportion and stir, let stand for 18-24 hours until transparent, then add an osmotic pressure regulator, and finally add the butenafine inclusion compound in the prescribed proportion prepared in step S1 and stir to dissolve to obtain an ophthalmic butenafine inclusion compound hydrogel.

8. The preparation method according to claim 7, characterized in that: The filtration in step S1 is performed by filtering using a 0.3-0.5 μm microporous filter membrane.

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