Externally applied medicine for treating and preventing onychomycosis as well as preparation method and application of externally applied medicine

Abstract

The invention provides an external medicine for treating and preventing onychomycosis as well as a preparation method and application thereof, and the external medicine is prepared from the following raw materials in parts by mass: 3-7% of ethyl acetate, 80-90% of squalane, 3-7% of octanoic acid and 3-7% of origanum vulgare leaf extract. The preparation method of the externally applied medicine comprises the following steps: uniformly stirring and mixing the caprylic acid, the origanum vulgare leaf extract, the squalane and the ethyl acetate to obtain the externally applied medicine. The external medicine can be prepared into spray or patch, so that the compliance of patients is improved, and the use is convenient. The external medicine prepared by the invention can fundamentally prevent and treat leuconychia caused by various fungi, and has the advantages of good curative effect, short treatment course, strong permeability, low recurrence rate, high safety, low cost and good application prospect.

Description

Technical Field

[0001] The present invention relates to the technical field of medicine preparation, and in particular to an external medicine for treating and preventing onychomycosis, and a preparation method and application thereof. Background Art

[0002] Onychomycosis, also known as onychomycosis, is a fungal disease that occurs on the nails and under the nails. The pathogenic fungi that cause this disease include dermatophytes, yeasts, yeast-like fungi, and Candida, etc. The damage to the nail caused by them is slightly different, and they are collectively referred to as onychomycosis in clinical practice. The main reasons why onychomycosis is difficult to treat are as follows: 1. Nail structure: The dense structure of the nail plate makes it difficult for traditional drugs to penetrate, providing a natural barrier for the reproduction of pathogens and a breeding ground for parasitism. 2. Fungal diversity: It is difficult for a single drug to be effective against all fungi. 3. Insensitivity to drugs: Fungal bodies in a dormant state are extremely poorly sensitive or not sensitive at all to general antifungal drugs, and the incubation period is particularly long. Once the medication is over, the spores will germinate, grow, and reproduce when encountering a suitable environment. 4. Repeated infection due to athlete's foot: In most cases, athlete's foot (tinea pedis) is first caused by onychomycosis, and of course, athlete's foot (tinea pedis) or tinea manuum can also be caused by onychomycosis.

[0003] At present, the means of treatment include surgical nail removal, oral medication, and topical medication. Surgical nail removal has a large wound, bleeding, easy to cause infection, and damage to the nail bed. It is necessary to change the dressing frequently, and it is generally painful after the operation, which causes great inconvenience to daily life and easily re-infects and relapses. Oral medication simply takes antifungal drugs (Sporanox, Terbinaquinol hydrochloride, Fluconazole, etc.), but this type of drug must reach the deck where the fungus is parasitic to play an antibacterial effect, and the solubility in water is low and slow, and the absorption and metabolism are very fast, so the dosage is large, the medication time is long, and it is easy to cause normal flora imbalance. Oral medication has side effects that can seriously damage the liver and kidneys, and it is easy to relapse after stopping the drug. The penetration of topical drugs to the deck is poor, and it cannot penetrate the protective layer on the surface of the nail to penetrate into the subungual or deep layer of the deck to kill the fungus, so the drug effect is not obvious after medication and the effect is slow, the drug effect time is short, it is difficult to cure, and it is easy to relapse. Therefore, it is necessary to develop an external medicine with good efficacy, short treatment course, strong permeability, low recurrence, small side effects and low cost to fundamentally treat onychomycosis caused by various fungi. Summary of the invention

[0004] In view of this, the present invention provides an external medicine for treating and preventing onychomycosis, and a preparation method and application thereof.

[0005] The technical solution of the present invention is achieved in this way:

[0006] An external medicine for treating and preventing onychomycosis consists of the following raw materials in parts by mass: 3%-7% ethyl acetate, 80%-90% squalane, 3%-7% octanoic acid, and 3%-7% oregano leaf extract.

[0007] Furthermore, the invention is composed of the following raw materials in parts by mass: 5% ethyl acetate, 85% squalane, 5% caprylic acid, and 5% oregano leaf extract.

[0008] Furthermore, the preparation method of the oregano leaf extract comprises:

[0009] (1) crushing fresh oregano leaves, adding them to a n-hexane-ethanol aqueous two-phase system, and ultrasonically extracting them to obtain a first filter residue and a first filtrate;

[0010] (2) adding the first filter residue into an ethanol-ammonium sulfate aqueous two-phase system and performing microwave extraction to obtain a second filter residue and a second filtrate;

[0011] (3) combining the first filtrate and the second filtrate, and concentrating under reduced pressure to obtain an oregano leaf extract.

[0012] Furthermore, in step (1), the solid-liquid ratio of the oregano leaves to the n-hexane-ethanol aqueous two-phase system is 1:10-20 g / ml; the concentration of the n-hexane-ethanol aqueous two-phase system is 40%-60% v / v; and the volume ratio of n-hexane to ethanol in the n-hexane-ethanol aqueous two-phase system is 1-3:1.

[0013] Furthermore, in step (1), the ultrasonic extraction is carried out at 40-50° C. and a power of 300-400 W for 1-3 h.

[0014] Furthermore, in step (2), the solid-liquid ratio of the first filter residue to the ethanol-ammonium sulfate two-phase aqueous system is 1:6-8 g / ml; the concentration of the ethanol-ammonium sulfate two-phase aqueous system is 50%-70% v / v; and the volume ratio of ethanol to ammonium sulfate in the ethanol-ammonium sulfate two-phase aqueous system is 1:2-3.

[0015] Furthermore, in step (2), the microwave extraction is carried out by microwave heating at 50-60° C. and a power of 400-500 W for 30-50 min.

[0016] A method for preparing an external medicine for treating and preventing onychomycosis comprises the following specific steps: uniformly mixing caprylic acid, oregano leaf extract, squalane and ethyl acetate to obtain the target external medicine.

[0017] Use of any one of the external medicines described in the invention in preparing medicines for treating and preventing onychomycosis.

[0018] Furthermore, the dosage form of the drug is a spray or a patch.

[0019] Compared with the prior art, the present invention has the following beneficial effects:

[0020] 1. The external medicine prepared by the present invention fundamentally prevents and treats onychomycosis caused by various fungi, has good efficacy, short treatment course, strong permeability, low recurrence rate, high safety, low cost and good application prospect.

[0021] 2. The squalane and oregano leaf extract of the present invention cooperate to resist fungal infection and inflammation, have a broad spectrum of bactericidal ability for various bacteria and fungi, enhance the effect of preventing and treating onychomycosis, and have no side effects on the human body. Adding squalane to the external medicine can not only soften keratin, but also make squalane and oregano leaf extract better penetrate the nail bed, achieve the effect of preventing and treating onychomycosis, and can also repair the epidermis, heal the wound surface, form a natural protective film, and promote the growth of new nails. The ethyl acetate of the present invention, as a solvent, can not only effectively dissolve squalane, squalane and oregano leaf extract, but also improve the utilization of squalane and oregano leaf extract, so that it can quickly penetrate into the fungal infection site, and achieve rapid and efficient treatment.

[0022] 3. The preparation process of the oregano leaf extract of the present invention utilizes "ultrasonic extraction of n-hexane-ethanol aqueous two-phase system + microwave extraction of ethanol-ammonium sulfate aqueous two-phase system", which can fully and highly selectively extract the active substances of the oregano leaves. DETAILED DESCRIPTION

[0023] In order to better understand the technical content of the present invention, specific embodiments are provided below to further illustrate the present invention.

[0024] Unless otherwise specified, the experimental methods used in the embodiments of the present invention are all conventional methods.

[0025] Unless otherwise specified, the materials, reagents, etc. used in the embodiments of the present invention can be obtained from commercial sources.

[0026] Example 1

[0027] An external medicine for treating and preventing onychomycosis consists of the following raw materials in parts by mass: 5% ethyl acetate, 85% squalane, 5% octanoic acid and 5% oregano leaf extract.

[0028] The method for preparing oregano leaf extract comprises:

[0029] (1) crushing fresh oregano leaves at a solid-liquid ratio of 1:15 g / ml, adding them to a 50% v / v n-hexane-ethanol aqueous two-phase system (the volume ratio of n-hexane to ethanol is 2:1), and ultrasonically extracting them at 45° C. and 350 W for 2 h to obtain a first filter residue and a first filtrate;

[0030] (2) adding the first filter residue to a 60% v / v ethanol-ammonium sulfate aqueous two-phase system (the volume ratio of ethanol to ammonium sulfate is 1:2.5) at a solid-liquid ratio of 1:7 g / ml, and heating the mixture in a microwave oven at 55° C. and a power of 450 W for 40 min to obtain a second filter residue and a second filtrate;

[0031] (3) combining the first filtrate and the second filtrate, and concentrating under reduced pressure to obtain an oregano leaf extract.

[0032] The preparation method of the external medicine for treating and preventing onychomycosis comprises the following specific steps: uniformly mixing caprylic acid, oregano leaf extract, squalane and ethyl acetate to obtain the target external medicine.

[0033] Example 2

[0034] An external medicine for treating and preventing onychomycosis consists of the following raw materials in parts by mass: 7% ethyl acetate, 87% squalane, 3% octanoic acid and 3% oregano leaf extract.

[0035] The method for preparing oregano leaf extract comprises:

[0036] (1) crushing fresh oregano leaves at a solid-liquid ratio of 1:10 g / ml, adding them to a 40% v / v n-hexane-ethanol aqueous two-phase system (the volume ratio of n-hexane to ethanol is 1:1), and ultrasonically extracting them at 40° C. and 300 W for 1 h to obtain a first filter residue and a first filtrate;

[0037] (2) adding the first filter residue to a 50% v / v ethanol-ammonium sulfate aqueous two-phase system (the volume ratio of ethanol to ammonium sulfate is 1:2) at a solid-liquid ratio of 1:6 g / ml, and heating the mixture in a microwave oven at 50° C. and a power of 400 W for 30 min to obtain a second filter residue and a second filtrate;

[0038] (3) combining the first filtrate and the second filtrate, and concentrating under reduced pressure to obtain an oregano leaf extract.

[0039] The preparation method of the external medicine for treating and preventing onychomycosis comprises the following specific steps: uniformly mixing caprylic acid, oregano leaf extract, squalane and ethyl acetate to obtain the target external medicine.

[0040] Example 3

[0041] An external medicine for treating and preventing onychomycosis consists of the following raw materials in parts by mass: 3% ethyl acetate, 83% squalane, 7% caprylic acid and 7% oregano leaf extract.

[0042] The method for preparing oregano leaf extract comprises:

[0043] (1) crushing fresh oregano leaves according to a solid-liquid ratio of 1:20 g / ml, adding them to a 60% v / v n-hexane-ethanol aqueous two-phase system (the volume ratio of n-hexane to ethanol is 3:1), and ultrasonically extracting them at 50° C. and 400 W for 3 h to obtain a first filter residue and a first filtrate;

[0044] (2) adding the first filter residue to an ethanol-ammonium sulfate aqueous two-phase system with a concentration of 70% v / v (the volume ratio of ethanol to ammonium sulfate is 1:3) at a solid-liquid ratio of 1:8 g / ml, and heating the mixture in a microwave at 60° C. and a power of 500 W for 50 min to obtain a second filter residue and a second filtrate;

[0045] (3) combining the first filtrate and the second filtrate, and concentrating under reduced pressure to obtain an oregano leaf extract.

[0046] The preparation method of the external medicine for treating and preventing onychomycosis comprises the following specific steps: uniformly mixing caprylic acid, oregano leaf extract, squalane and ethyl acetate to obtain the target external medicine.

[0047] Comparative Example 1

[0048] The difference from Example 1 is that the amount of raw materials is different; the rest is the same as Example 1.

[0049] The external medicine for treating and preventing onychomycosis in this comparative example is composed of the following raw materials in parts by mass: 15% ethyl acetate, 75% squalane, 2% octanoic acid, and 8% oregano leaf extract.

[0050] Comparative Example 2

[0051] The difference from Example 1 is that caprylic acid is missing, that is, the oregano leaf extract is 10%; the rest is consistent with Example 1.

[0052] The external medicine for treating and preventing onychomycosis in this comparative example is composed of the following raw materials in parts by mass: 5% ethyl acetate, 85% squalane, and 10% oregano leaf extract.

[0053] Among them, the preparation method of the external medicine for treating and preventing onychomycosis in this comparative example specifically comprises the following steps: stirring and mixing oregano leaf extract, squalane and ethyl acetate to obtain the target external medicine.

[0054] Comparative Example 3

[0055] The difference from Example 1 is that the oregano leaf extract is missing, that is, the octanoic acid is 10%; the rest is the same as Example 1.

[0056] The external medicine for treating and preventing onychomycosis in this comparative example is composed of the following raw materials in parts by mass: 5% ethyl acetate, 85% squalane, and 10% octanoic acid.

[0057] The preparation method of the external medicine for treating and preventing onychomycosis in this comparative example specifically comprises the following steps: uniformly mixing caprylic acid, squalane and ethyl acetate to obtain the target external medicine.

[0058] Comparative Example 4

[0059] The difference from Example 1 is that squalane is missing, that is, ethyl acetate is 90%; the rest is the same as Example 1.

[0060] The external medicine for treating and preventing onychomycosis in this comparative example is composed of the following raw materials in parts by mass: 90% ethyl acetate, 5% octanoic acid, and 5% oregano leaf extract.

[0061] The preparation method of the external medicine for treating and preventing onychomycosis in this comparative example specifically comprises the following steps: uniformly mixing caprylic acid, oregano leaf extract and ethyl acetate to obtain the target external medicine.

[0062] Comparative Example 5

[0063] The difference from Example 1 is that the preparation method of the oregano leaf extract is ethanol ultrasonic extraction; the rest is consistent with Example 1.

[0064] The preparation method of the oregano leaf extract of the comparative example comprises: crushing fresh oregano leaves according to a solid-liquid ratio of 1:15 g / ml, adding a 50% v / v ethanol solution, ultrasonically extracting for 2 hours at 45° C. and a power of 350 W, collecting the extract, and concentrating under reduced pressure to obtain the oregano leaf extract.

[0065] Antibacterial test

[0066] 1. Fungi: Trichophyton rubrum, Trichophyton mentagrophytes, Microsporum canis, Epidermophyton floccosum, Escherichia coli, Staphylococcus aureus, Candida albicans and Trichophyton furfur;

[0067] 2. Experimental groups: control group (sterile water) and experimental group (Examples 1-3, Comparative Examples 1-5), commercially available group (amorolfine hydrochloride liniment);

[0068] 3. Experimental method: First, inject the bacterial solution (the 24-hour slant culture of the test bacteria was washed with PBS to make a bacterial suspension) and agar medium into a flat plate, mix and wait for coagulation, use a sterile hole puncher to punch 6 holes with a diameter of 6 mm on the agar plate, pick out the agar in the holes, and then drop the topical medicine of the experimental group diluted 2 times (0.2 mL / hole), culture at 25°C for 24 hours, and observe the size of the inhibition zone and the effect on the activity of the fungus as shown in Table 1-2.

[0069] Table 1

[0070]

[0071]

[0072] Table 2

[0073]

[0074] As can be seen from Table 1-2, the external medicine prepared by Examples 1-3 of the present invention has a strong antibacterial and antifungal effect, achieves a good effect of preventing and treating onychomycosis, and is better than the onychomycosis external medicine (amorolfine hydrochloride liniment) of the commercially available group. Comparative Example 1 Compared with Example 1, the raw material dosage is different, resulting in a reduction in its antibacterial rate, and a decrease in the efficacy of preventing and treating onychomycosis; Comparative Example 2 lacks octanoic acid, and Comparative Example 3 lacks oregano leaf extract, resulting in a reduction in its antibacterial rate, and a decrease in the efficacy of preventing and treating onychomycosis. It can be concluded that the octanoic acid and oregano leaf extract of the present invention have a synergistic effect, jointly improve the effect of antifungal infection, have a broad spectrum of bactericidal ability for various bacteria and fungi, and enhance the effect of preventing and treating onychomycosis; Comparative Example 4 lacks squalane, and its antibacterial rate is reduced, and the efficacy of preventing and treating onychomycosis is reduced, indicating that the squalane of the present invention can not only soften keratin, but also make octanoic acid and oregano leaf extract better penetrate the nail bed, achieve the effect of preventing and treating onychomycosis, but also can repair the epidermis, heal the wound, form a natural protective film, and promote the growth of new nails. The preparation method of the oregano leaf extract of Comparative Example 5 is ethanol ultrasonic extraction, and its antibacterial rate is reduced, and the efficacy of preventing and treating onychomycosis is reduced, indicating that the preparation method of the oregano leaf extract of the present invention utilizes "n-hexane-ethanol two-phase aqueous system ultrasonic extraction + ethanol-ammonium sulfate two-phase aqueous system microwave extraction", which can fully and highly selectively extract the active substances of oregano leaves, thereby enhancing the broad-spectrum bactericidal ability of various bacteria and fungi.

[0075] Application Example 1-Spray

[0076] The external medicine prepared in Example 1 (10 wt%), polyethylene glycol (3 wt%), preservative (0.5 wt%) and the remainder of water, totaling 100%, were mixed evenly, filled in a spray bottle, and filled with pressure (50 KPa) to prepare a spray.

[0077] Application Example 2-Patch

[0078] The external medicine prepared in Example 1 (10 wt%), cellulose (5 wt%), preservative (0.5 wt%) and the remaining filler, totaling 100%, were mixed evenly and coated evenly on a polyethylene film as a base material to prepare a patch.

[0079] Clinical Research

[0080] 150 patients with onychomycosis were randomly divided into 3 groups, 50 in each group. The first group sprayed the spray of Application Example 1 on the onychomycosis twice a day; the second group applied the patch of Application Example 2 on the onychomycosis and changed the dressing twice a day; the third group applied the commercially available amorolfine hydrochloride ointment on the onychomycosis and changed the dressing twice a day; any other treatment was stopped during the treatment of the three groups, and the efficacy was observed after 1 week.

[0081] Treatment standard: Cured: The diseased nail disappears completely, most of the new nail grows out, there is no itching, and the new nail plate is as smooth and bright as the disease-free nail plate;

[0082] Effective: Part of the nail grows out, the brittleness improves, there is no separation from the nail bed, no cavities, and no itching;

[0083] Ineffective: There is no significant change in symptoms after medication compared to before treatment.

[0084] The statistical results are shown in Table 3.

[0085] Table 3

[0086]

[0087]

[0088] As can be seen from Table 3, the external-use pharmaceutical preparations of Application Examples 1-2 of the present invention have better efficacy than the amorolfine hydrochloride ointment of the commercially available group, and the external-use pharmaceutical preparations of the present invention have good efficacy, short treatment course, low recurrence rate, high safety, and good application prospects.

[0089] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the protection scope of the present invention.

Claims

1. An external medicine for treating and preventing onychomycosis, characterized in that: The invention is composed of the following raw materials in terms of mass: 3%-7% ethyl acetate, 80%-90% squalane, 3%-7% octanoic acid and 3%-7% oregano leaf extract.

2. The external medicine for treating and preventing onychomycosis according to claim 1, characterized in that: The invention is composed of the following raw materials in parts by mass: 5% ethyl acetate, 85% squalane, 5% caprylic acid and 5% oregano leaf extract.

3. The external medicine for treating and preventing onychomycosis according to claim 1, characterized in that: The preparation method of the oregano leaf extract comprises: (1) crushing fresh oregano leaves, adding them to a n-hexane-ethanol aqueous two-phase system, and ultrasonically extracting them to obtain a first filter residue and a first filtrate; (2) adding the first filter residue into an ethanol-ammonium sulfate aqueous two-phase system and performing microwave extraction to obtain a second filter residue and a second filtrate; (3) combining the first filtrate and the second filtrate, and concentrating under reduced pressure to obtain an oregano leaf extract.

4. The external medicine for treating and preventing onychomycosis according to claim 3, characterized in that: In step (1), the solid-liquid ratio of the oregano leaves to the n-hexane-ethanol aqueous two-phase system is 1:10-20 g / ml; the concentration of the n-hexane-ethanol aqueous two-phase system is 40%-60% v / v; and the volume ratio of n-hexane to ethanol in the n-hexane-ethanol aqueous two-phase system is 1-3:

1.

5. The external medicine for treating and preventing onychomycosis according to claim 3, characterized in that: In step (1), the ultrasonic extraction is carried out at 40-50°C and a power of 300-400W for 1-3h.

6. The external medicine for treating and preventing onychomycosis according to claim 3, characterized in that: In step (2), the solid-liquid ratio of the first filter residue to the ethanol-ammonium sulfate two-phase aqueous system is 1:6-8 g / ml; the concentration of the ethanol-ammonium sulfate two-phase aqueous system is 50%-70% v / v; and the volume ratio of ethanol to ammonium sulfate in the ethanol-ammonium sulfate two-phase aqueous system is 1:2-3.

7. The external medicine for treating and preventing onychomycosis according to claim 3, characterized in that: In step (2), the microwave extraction is carried out by microwave heating at 50-60° C. and a power of 400-500 W for 30-50 min.

8. The method for preparing an external medicine for treating and preventing onychomycosis according to claim 1, characterized in that: The specific steps include: stirring and mixing caprylic acid, oregano leaf extract, squalane and ethyl acetate to obtain the target external medicine.

9. Use of the external medicine according to any one of claims 1 to 7 in the preparation of a medicine for treating and preventing onychomycosis.

10. The use according to claim 9, characterized in that The dosage form of the medicine is a spray or a patch.

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