Liposome preparation capable of expelling parasites, inhibiting bacteria and killing mites as well as preparation method and application of liposome preparation

Abstract

The invention relates to a liposome preparation for expelling parasites, inhibiting bacteria and killing mites as well as a preparation method and application thereof, and belongs to the technical field of daily chemicals. The liposome preparation for expelling parasites, inhibiting bacteria and killing mites provided by the invention comprises a liposome content, a liposome wall material, a liposome curing agent and water, and is prepared from the following components in percentage by mass: 0.1 to 10 percent of bactericide, 0.1 to 10 percent of insecticide, 1 to 40 percent of bacteriostatic agent, 0.1 to 10 percent of efflux pump inhibitor, 0.1 to 10 percent of liposome wall material, 0.01 to 1 percent of liposome curing agent and the balance of water. The dosage form is lipidosome, the particle size is small, the permeability is very high, and the active ingredients can be assisted to take effect better; the components are low-toxicity or natural-source components with relatively high safety; the sustained-release effect is achieved, burst release of active ingredients is prevented, and safer and longer-acting application performance is provided.

Description

Technical Field

[0001] The present invention relates to the technical field of daily chemicals, and in particular to an insect repellent, antibacterial and mite removing liposome preparation and a preparation method and application thereof. Background Art

[0002] Mites are closely related to human health. Studies have found that mites can invade the skin, causing "rosacea" or demodectic mange, allergies, urinary tract mite disease, lung mite disease, intestinal mite disease and scabies, which seriously endanger human health. Therefore, household mite removal products are increasingly attracting the attention and use of consumers.

[0003] At present, among the wide variety of mite removal products, daily chemical products account for 80%-90%, including mite removal spray, mite removal medicine packs, mite removal laundry detergent, etc.; in the field of textile printing and dyeing, by adding textile auxiliaries with mite removal effect, the fabric has mite removal function and is used in products such as clothing, bed sheets, sofa covers, etc.

[0004] However, looking at the mite removal products, either they contain chemical mite removal agents, which have a strong odor, are harmful to the human body, and are not effective for a long time. Or they are plant-based mite removal agents, which are less harmful to the human body, but often have average effects. How to balance the safety and effectiveness of mite removal products is an industry pain point that needs to be solved urgently, and it is also the core concern of consumers; the above problems are also common in commercially available repellent products, such as insecticides, mosquito repellents, antibacterial agents, etc.

[0005] Therefore, developing a preparation that has good combined effects of insect repelling, antibacterial and mite removal and is less harmful to the skin has high application value in the field of daily chemicals. Summary of the invention

[0006] The purpose of the present invention is to overcome the shortcomings of the prior art and provide an insect repellent, antibacterial, and mite removal liposome preparation and its preparation method and application. The insect repellent, antibacterial, and mite removal liposome preparation provided by the present invention has strong permeability, can achieve low-dose high effect, high safety, and has a sustained-release effect to provide long-term application performance, and has high application value in the field of daily chemicals.

[0007] To achieve the above object, the technical solution adopted by the present invention is:

[0008] In a first aspect, the present invention provides an insect repellent, antibacterial, and mite removal liposome preparation, comprising liposome contents, liposome wall materials, liposome curing agents, and water, wherein the liposome contents include bactericides, insecticides, antibacterial agents, and efflux pump inhibitors; the antibacterial agent is a polyol, and the liposome curing agent is a C16-C22 fatty acid;

[0009] The insect repellent, antibacterial and mite removing liposome preparation comprises the following components by mass percentage: 0.1-10% bactericide, 0.1-10% insecticide, 1-40% antibacterial agent, 0.1-10% efflux pump inhibitor, 0.1-10% liposome wall material, 0.01-1% liposome curing agent and the balance water.

[0010] The present invention provides a composite functional agent with the effects of anthelmintic, antibacterial and mite removal, and the dosage form is liposome; the contents of the liposome include bactericide, insecticide, antibacterial agent and efflux pump inhibitor, wherein the fat-soluble components can be contained between the bilayers of the wall material, and the water-soluble components can be contained inside the bilayers of the liposome; in addition, a liposome curing agent is added to fix the shape and form a more stable preparation.

[0011] Among them, bactericides provide a broad-spectrum bactericidal effect; insecticides have the effects of repelling insects, killing insects, and removing mites, and are relatively safe; in addition to providing a broad-spectrum antibacterial effect and enhancing antibacterial and antimicrobial effects, polyol antibacterial agents can also play the following roles when used in combination with other components: prevent biomacromolecules from forming biofilms outside bacterial cells, thereby reducing bacterial resistance and achieving antibacterial and antimicrobial enhancement effects; they can also be used as solvents to dissolve liposome wall materials. Efflux pump inhibitors can interfere with the assembly of efflux pumps on bacterial cell membranes, block the energy source of efflux pumps, and hinder substrates from directly passing through efflux channels, thereby directly interfering with the efflux function of bacteria, inhibiting bacterial activity, and achieving antibacterial, antibacterial, and insecticidal enhancement effects.

[0012] The liposome wall material can encapsulate the above-mentioned bactericides, pesticides, antibacterial agents and efflux pump inhibitors, and is the structural basis for the composite agent of the present invention to construct the drug-carrying liposome. Gram-negative bacteria contain a lot of bacterial outer membranes that are difficult for drugs to penetrate, which is the main reason why most antibacterial drugs are ineffective against Gram-negative bacteria. The bacterial cell membrane is mainly composed of lipids (mainly phospholipids and cholesterol), etc. When the liposome wall material in the present invention contacts bacteria, it can inactivate the permeability barrier of the bacterial outer membrane. The liposome wall material can easily carry drugs into bacteria, thereby reducing the drug resistance of bacteria, allowing each component to play a role directly in bacteria, and achieving a synergistic effect.

[0013] In addition, the liposome curing agent introduced in the present invention can adjust the appearance of the liposome wall material and improve the sustained release effect of the liposome complex of the present invention. The liposome curing agent used in the present invention is a fatty acid substance that can solidify liquid fat. The fatty acid chain length is relatively short, the chain length is between C16-C22, and the mutual force between them is relatively weak; when the temperature drops, the mutual force between molecules gradually increases, so that the fat molecules begin to gather together to form small crystal nuclei, and these crystal nuclei will continue to grow, and finally form a continuous and interconnected solid structure, so that the liquid fat is converted into solid fat. The role of water is to dissolve and disperse the components.

[0014] The liposome encapsulation and solidification strategy adopted in the present invention can significantly improve the insect repellent, antibacterial and mite removal effects of the composite agent. If the liposome dosage form is not adopted or the liposome solidification is not performed, the overall performance will be significantly reduced. The ratio of the liposome wall material and the liposome solidifying agent also has a significant impact on the insect repellent, antibacterial and mite removal effects of the present invention. Only under a specific ratio can a better insect repellent, antibacterial and mite removal effect be comprehensively achieved.

[0015] The anthelmintic, antibacterial and mite-removing liposome preparation provided by the present invention can form a drug-carrying liposome structure with a small particle size and strong permeability, which can help the active ingredients to take effect better and achieve the purpose of low dose and high effect; the anthelmintic, antibacterial and mite-removing liposome preparation provided by the present invention contains antibacterial, antibacterial and insecticidal synergistic ingredients, which can reduce receptor resistance; at the same time, the present invention has a sustained release effect, which can prevent the explosive release of active ingredients and provide safer and longer-lasting application performance.

[0016] Preferably, the insect repellent, antibacterial and mite removal liposome preparation comprises the following components by mass percentage: 0.5-10% bactericide, 1-8% insecticide, 20-35% antibacterial agent, 1-5% efflux pump inhibitor, 2-3% liposome wall material, 0.2-0.5% liposome curing agent and the remainder water.

[0017] As a preferred embodiment, the insect repellent, antibacterial and mite removal liposome preparation includes the following components, by mass percentage: 5-10% bactericide, 5-8% insecticide, 30-35% antibacterial agent, 3-5% efflux pump inhibitor, 3% liposome wall material, 0.3-0.5% liposome curing agent and the remainder water.

[0018] Preferably, the bactericide is at least one of diclosan, phenoxyethanol, polyhexamethylene hydrochloride, sodium benzoate, potassium sorbate, methylparaben, propylparaben, methylisothiazolinone, methylchloroisothiazolinone, DMDM ​​hydantoin and chlorphenesin.

[0019] Preferably, the insecticide is at least one of natural menthanediol, pyrethrin, pepper extract, onion extract, orange peel extract, garlic extract, honeysuckle extract, sophora flavescens extract, ginkgo extract, derris extract, artemisia annua extract, castor seed extract, and datura extract.

[0020] Preferably, the antibacterial agent is at least one of glycerol, ethanol, propylene glycol, butylene glycol, pentanediol and hexylene glycol.

[0021] The bactericide, bacteriostatic agent and insecticide provided by the present invention are all low-toxic or natural ingredients, have high safety, little harm to the skin, are non-irritating to the skin, and are more suitable for the field of daily chemicals.

[0022] Preferably, the efflux pump inhibitor is at least one of glycyrrhizic acid, methyl pyrrolidone, artesunate, antisense nucleic acid, oligonucleotide, and amikacin.

[0023] Preferably, the liposome wall material is at least one of lecithin, hydrogenated lecithin, hydroxylated lecithin, hydrogenated lysolecithin, lysolecithin, and soybean lecithin.

[0024] Preferably, the liposome solidifying agent is at least one of palmitic acid, linoleic acid, linolenic acid, oleic acid, stearic acid, arachidic acid, and behenic acid.

[0025] In a second aspect, the present invention provides a method for preparing the above-mentioned insect repellent, antibacterial and mite removing liposome preparation, comprising the following steps:

[0026] (1) dispersing a fungicide, an insecticide and an efflux pump inhibitor in deionized water, heating and keeping the water warm, and mixing the water to obtain a phase A;

[0027] (2) dissolving the liposome wall material in the antibacterial agent, heating and keeping warm, and mixing to obtain phase B;

[0028] (3) adding the phase B dropwise into the phase A under heating, insulation and stirring to obtain phase C;

[0029] The heating and heat preservation temperature is 35-65°C;

[0030] (4) homogenizing the phase C to obtain phase D, then heating the phase to 45-75° C., dispersing the liposome solidifying agent in the phase D, and cooling the phase to 15-30° C. to obtain the insect repellent, antibacterial, and mite removal liposome preparation.

[0031] The anthelmintic, antibacterial and de-mite liposome preparation prepared by the above preparation method can prepare an anthelmintic, antibacterial and de-mite liposome preparation with stable dosage form and good sustained-release effect, so that the drug-carrying liposome can release the effective components stably for a long time and achieve longer-lasting application performance.

[0032] Preferably, the cooling time is 0.5-3h.

[0033] In a third aspect, the present invention provides the application of the above-mentioned insect repellent, antibacterial and mite removal liposome preparation in the field of daily chemicals.

[0034] The insect repellent, antibacterial and mite removal liposome preparation provided by the present invention has a safe and long-lasting comprehensive insect repellent, antibacterial and mite removal effect, and can achieve high effect at a low dose. In the field of daily chemical products, it can be further added and applied to a variety of products, such as laundry detergent, pet shampoo, etc., and has a high application value.

[0035] Compared with the prior art, the present invention has the following beneficial effects:

[0036] The present invention provides a composite agent with good insect repellent, antibacterial and mite removal effects by introducing liposome wall materials and liposome curing agents and limiting the functional components and proportions. The components are all of low toxicity or natural origin, highly safe and non-irritating to the skin; the composite agent also has strong permeability and contains antibacterial and antibacterial, insecticidal and synergistic ingredients, which can reduce receptor resistance and comprehensively achieve low-dose and high-effect; at the same time, the composite agent has a stable and long-lasting sustained-release effect, which can prevent the explosive release of active ingredients, provide safe and long-lasting application performance, and is more suitable for the use needs of daily chemical products, and has a high application value. BRIEF DESCRIPTION OF THE DRAWINGS

[0037] Figure 1 The transmission electron micrograph of the insect repellent, antibacterial and mite removal liposome preparation in Example 1;

[0038] Figure 2 The particle size distribution diagram of the insect repellent, antibacterial and mite removal liposome preparation in Example 1;

[0039] Figure 3 The minimum inhibitory concentration (MIC) results of the liposome preparations for insect repelling, antibacterial and mite removal in the examples and comparative examples are shown;

[0040] Figure 4 A schematic diagram of a sticky board used in a mite removal test in an effect example;

[0041] Figure 5 The graph is a result of mite removal rate of different concentration gradients of the anthelmintic, antibacterial and mite removal liposome preparations in the examples and comparative examples;

[0042] Figure 6 The graph is a release rate curve of the active ingredients of the insect repellent, antibacterial and mite removal liposome preparations in water in Example 1 and Comparative Examples 1 and 2. DETAILED DESCRIPTION

[0043] In order to better illustrate the purpose, technical scheme and advantages of the present invention, the present invention will be further described below in conjunction with specific examples. The test methods used in the following examples are conventional methods unless otherwise specified; the materials and reagents used are reagents and materials that can be obtained from commercial channels unless otherwise specified.

[0044] Example 1

[0045] An embodiment of the insect repellent, antibacterial and mite removal liposome preparation of the present invention, the insect repellent, antibacterial and mite removal liposome preparation described in this embodiment is composed of the following components by mass percentage: 1% bactericide, 1% insecticide, 1% efflux pump inhibitor, 20% antibacterial agent, 2% liposome wall material, 0.2% liposome curing agent and the remainder water.

[0046] The preparation method of the insect repellent, antibacterial and mite removal liposome preparation described in this embodiment is as follows:

[0047] (1) Weigh 1 g of diclosan, 1 g of natural menthyl glycol, and 1 g of glycyrrhizic acid and disperse them in 74.8 g of deionized water, heat at 45° C., and maintain a stirring speed of 500 rpm / min to obtain phase A1;

[0048] (2) Weigh 2 g of soybean lecithin and uniformly dissolve it in 20 g of propylene glycol, and heat it at 45°C to obtain phase B1;

[0049] (3) heating at 45°C and stirring at 500 rpm / min, adding phase B1 dropwise into phase A1 to obtain phase C1;

[0050] (4) homogenizing phase C1 by a high-pressure homogenizer at a homogenization pressure of 1000 bar to obtain phase D1;

[0051] (5) The temperature of phase D1 was raised to 55°C, 0.2 g of palmitic acid was evenly dispersed therein, and the stirring speed was maintained at 50 rpm / min. The temperature was slowly lowered to 30°C for 0.5 h to obtain the insect repellent, antibacterial and mite removal liposome preparation, hereinafter referred to as "Example 1".

[0052] The transmission electron microscopy image of the insect repellent, antibacterial and mite removal liposome preparation of Example 1 is as follows: Figure 1 , the particle size distribution diagram is as follows Figure 2 It can be seen that this example successfully prepared an insect repellent, antibacterial, and mite-removing liposome preparation, the average particle size of which was 95 nm.

[0053] Example 2

[0054] An embodiment of the insect repellent, antibacterial and mite removal liposome preparation of the present invention, the insect repellent, antibacterial and mite removal liposome preparation described in this embodiment is composed of the following components by mass percentage: 5% bactericide, 5% insecticide, 3% efflux pump inhibitor, 30% antibacterial agent, 3% liposome wall material, 0.3% liposome curing agent and the remainder water.

[0055] The preparation method of the insect repellent, antibacterial and mite removal liposome preparation described in this embodiment is as follows:

[0056] (1) Weigh 5 g of sodium benzoate, 5 g of natural menthyl glycol, and 3 g of methyl pyrrolidone and disperse them in 53.7 g of deionized water, heat at 55° C., and maintain a stirring speed of 800 rpm / min to obtain phase A2;

[0057] (2) Weigh 3 g of soybean lecithin and uniformly dissolve it in 30 g of butanediol and heat at 45° C. to obtain phase B2;

[0058] (3) heating at 55°C and stirring at 800 rpm / min, adding phase B1 dropwise into phase A2 to obtain phase C2;

[0059] (4) homogenizing the C2 phase by a high-pressure homogenizer at a homogenization pressure of 2000 bar to obtain the D2 phase;

[0060] (5) The temperature of phase D2 was raised to 65°C, 0.3 g of linoleic acid was evenly dispersed therein, and the stirring speed was maintained at 70 rpm / min. The temperature was slowly lowered to 25°C for 1.5 h to obtain the insect repellent, antibacterial and mite removal liposome preparation, hereinafter referred to as "Example 2".

[0061] Example 3

[0062] An embodiment of the insect repellent, antibacterial and mite removal liposome preparation of the present invention, the insect repellent, antibacterial and mite removal liposome preparation described in this embodiment is composed of the following components by mass percentage: 10% bactericide, 8% insecticide, 5% efflux pump inhibitor, 30% antibacterial agent, 3% liposome wall material, 0.5% liposome curing agent and the remainder water.

[0063] The preparation method of the insect repellent, antibacterial and mite removal liposome preparation described in this embodiment is as follows:

[0064] (1) Weigh 10 g of potassium sorbate, 8 g of pyrethrin, and 5 g of glycyrrhizic acid, dissolve or disperse them in 43.5 g of deionized water, heat at 65° C., and maintain a stirring speed of 1000 rpm / min to obtain phase A3;

[0065] (2) Weigh 3 g of hydrogenated lecithin and uniformly dissolve it in 30 g of glycerol, and heat it at 65° C. to obtain phase B3;

[0066] (3) heating at 65°C and stirring at 1000 rpm / min, adding phase B1 dropwise into phase A3 to obtain phase C3;

[0067] (4) homogenizing the C3 phase by a high-pressure homogenizer at a homogenization pressure of 2500 bar to obtain the D3 phase;

[0068] (5) The temperature of phase D3 was raised to 55°C, 0.5 g of stearic acid was evenly dispersed therein, and the stirring speed was maintained at 50 rpm / min. The temperature was slowly lowered to 15°C for 2 h to obtain the insect repellent, antibacterial and mite removal liposome preparation, hereinafter referred to as "Example 3".

[0069] Example 4

[0070] An embodiment of the anthelmintic, antibacterial and mite-removing liposome preparation of the present invention is different from Example 1 only in that the amount of glycyrrhizic acid is reduced to 0.2 g, and the amount of deionized water is correspondingly increased to keep the mass percentages of other components unchanged, that is, the efflux pump inhibitor is 0.2% by mass, to obtain an anthelmintic, antibacterial and mite-removing liposome preparation, hereinafter referred to as "Example 4".

[0071] Example 5

[0072] An embodiment of the insect repellent, antibacterial and mite removal liposome preparation of the present invention is different from Example 1 only in that the amount of soybean lecithin is reduced to 0.5g, the amount of palmitic acid is increased to 0.6g, and the amount of deionized water is adjusted accordingly to keep the mass percentages of other components unchanged, that is, the mass percentages are 0.5% of liposome wall material and 0.6% of liposome curing agent, to obtain an insect repellent, antibacterial and mite removal liposome preparation, hereinafter referred to as "Example 5".

[0073] Example 6

[0074] An embodiment of the insect repellent, antibacterial and mite removal liposome preparation of the present invention is different from Example 1 only in that the amount of palmitic acid is increased to 0.5 g, the amount of diclosan is reduced to 0.5 g, and the amount of deionized water is adjusted accordingly to keep the mass percentages of other components unchanged, that is, the mass percentages are 0.5% of liposome curing agent and 0.6% of bactericide, to obtain an insect repellent, antibacterial and mite removal liposome preparation, hereinafter referred to as "Example 6".

[0075] Comparative Example 1

[0076] Comparative Example 1 is based on Example 1, but without preparing into a liposome dosage form and removing the solidification step. The preparation process includes the following steps:

[0077] (1) Weigh 1 g of diclosan, 1 g of natural menthyl glycol, and 1 g of glycyrrhizic acid, dissolve or disperse them in 74.8 g of deionized water, heat at 45° C., and maintain a stirring speed of 500 rpm / min to obtain phase A4;

[0078] (2) Weigh 2 g of soybean lecithin and uniformly dissolve it in 20 g of propylene glycol, and heat it at 45° C. to obtain phase B4;

[0079] (3) Under heating at 45°C and maintaining a stirring speed of 500 rpm / min, phase B4 was added dropwise into phase A4 to obtain phase C4, hereinafter referred to as "Comparative Example 1".

[0080] Comparative Example 2

[0081] Comparative Example 2 is based on Example 1, but the curing step is removed, and the preparation process includes the following steps:

[0082] (1) Weigh 1 g of diclosan, 1 g of natural menthyl glycol, and 1 g of glycyrrhizic acid, dissolve or disperse them in 74.8 g of deionized water, heat at 45° C., and maintain a stirring speed of 500 rpm / min to obtain phase A5;

[0083] (2) Weigh 2 g of soybean lecithin and uniformly dissolve it in 20 g of propylene glycol, and heat it at 45° C. to obtain phase B5;

[0084] (3) heating at 45°C and stirring at 500 rpm / min, adding phase B5 dropwise into phase A5 to obtain phase C5;

[0085] (4) Phase C5 was homogenized by a high-pressure homogenizer at a homogenization pressure of 1000 bar to obtain phase D5, hereinafter referred to as "Comparative Example 2".

[0086] Comparative Example 3

[0087] Comparative Example 3 is based on Example 1, but liposomes are not formed. The preparation process includes the following steps:

[0088] (1) Weigh 1 g of diclosan, 1 g of natural menthyl glycol, and 1 g of glycyrrhizic acid and disperse them in 74.8 g of deionized water, heat at 45° C., and maintain a stirring speed of 500 rpm / min to obtain phase A6;

[0089] (2) Weigh 2 g of soybean lecithin and uniformly dissolve it in 20 g of propylene glycol, and heat it at 45° C. to obtain phase B6;

[0090] (3) heating at 45°C and stirring at 500 rpm / min, adding phase B6 dropwise into phase A6 to obtain phase C6;

[0091] (4) The temperature of phase C6 was raised to 55°C, 0.2 g of palmitic acid was evenly dispersed therein, and the stirring speed was maintained at 50 rpm / min. The temperature was slowly lowered to 30°C for 0.5 h to obtain phase D6, hereinafter referred to as "Comparative Example 3".

[0092] Comparative Example 4

[0093] The difference between Comparative Example 4 and Example 1 is that the amount of glycyrrhizic acid is reduced to 0.01 g, and the amount of deionized water is adjusted accordingly to keep the mass percentages of other components unchanged, that is, the mass percentage of the efflux pump inhibitor is 0.01%, and a sample is prepared, hereinafter referred to as "Comparative Example 4 sample".

[0094] Comparative Example 5

[0095] The difference between Comparative Example 5 and Example 1 is that the amount of soybean lecithin is reduced by 0.01 g, the amount of palmitic acid is increased by 1.5 g, and the amount of deionized water is adjusted accordingly to keep the mass percentages of other components unchanged, that is, the mass percentages are 0.01% liposome wall material and 1.5% liposome curing agent, to prepare a sample, hereinafter referred to as "Comparative Example 5 sample".

[0096] Effect Example 1

[0097] In order to explore the antibacterial efficacy of the anthelmintic, antibacterial and mite-removing liposome preparation provided by the present invention, the anthelmintic, antibacterial and mite-removing liposome preparations of the examples and the samples of the comparative examples were subjected to the following minimum inhibitory concentration MIC (%) test:

[0098] The test was carried out in accordance with the 2002 edition of the Ministry of Health's "Technical Specifications for Disinfection" - 2.1.8.3 Minimum Inhibitory Concentration Test (Agar Dilution Method). The test strains were Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, Candida albicans, and Aspergillus niger.

[0099] Preparation of culture medium: Use MH agar, prepared according to the instructions of the product, pH 7.2-7.4. For Neisseria gonorrhoeae, use GC agar base plus 1% additive; for other streptococci, use MH agar containing 5% (V / V) sheep blood.

[0100] Preparation of drug-containing agar plates: According to the experimental design, add the antimicrobial drugs of different concentrations that have been diluted in multiples to the MH agar that has been heated and dissolved and balanced in a 45-50℃ water bath, mix thoroughly and pour into sterilized plates, the agar thickness is 3-4mm. Usually, drug agar plates are prepared at a ratio of 1:9, and the drug concentration range is selected according to needs. The prepared drug-containing agar plates should be placed in a sealed plastic bag and stored in a refrigerator at 2-8℃ for 5 days.

[0101] Inoculum preparation and inoculation: Prepare a bacterial suspension with a concentration equivalent to 0.5 McFarland standard turbidimetric tube, dilute it 1:10, and use a multi-point inoculator to draw the prepared bacterial suspension (about 1-2μl) and inoculate it on the surface of the agar plate. The number of bacteria per point is about 104CFU, forming a plaque with a diameter of 5-8mm. After inoculation, incubate at 35℃ for 16-20h and observe the results.

[0102] Result judgment: Place the plate on a dark, non-reflective surface to judge the test endpoint, and the lowest drug concentration that inhibits bacterial growth is MIC. Slight bacterial growth can be seen on agar plates containing trimethoprim or sulfa, and the lowest drug concentration that inhibits more than 80% bacterial growth compared with the growth control is taken as the endpoint concentration. If more than two colonies grow on agar plates containing drug concentrations higher than the endpoint level, or if they do not grow on low-concentration drug agar plates but grow on high-concentration drug agar plates, the culture purity should be checked or the test should be repeated.

[0103] The test results of MIC are shown in Table 1 and Figure 3 As shown, the lower the MIC concentration, the better the antibacterial effect.

[0104] From Table 1, Figure 3 It can be seen that:

[0105] (1) The anthelmintic, antibacterial and mite-removing liposome preparations provided in the embodiments of the present invention all have good antibacterial effects, and the minimum inhibitory concentration MIC (%) is significantly lower than that of the samples in the comparative examples. Among them, the anthelmintic, antibacterial and mite-removing liposome preparation provided in Example 3 has the lowest antibacterial MIC and the best effect.

[0106] (2) Comparison of Example 1 and Comparative Examples 1-3: It can be seen that the MIC of Example 1 is significantly lower than that of Comparative Examples 1-3. In Comparative Example 2, no liposome curing agent is added and no liposome curing step is performed, resulting in a significant increase in MIC and a decrease in antibacterial effect. In Comparative Example 3, the components are the same but no liposomes are formed, the effect on bacterial cells is weakened, and the antibacterial effect is also significantly reduced. In Comparative Example 1, no liposomes are formed and no curing agent is added, the MIC is further increased, and the antibacterial effect is poor. This verifies that forming a liposome dosage form and further curing the liposomes can significantly improve the antibacterial effect and reduce the MIC.

[0107] (3) From the comparison between Example 1 and Comparative Example 5, it can be seen that the usage ratio of liposome wall material and liposome curing agent also affects the antibacterial effect and MIC. When too little liposome wall material and too much liposome curing agent are used in Comparative Example 5, the MIC is increased by about 50%, and the antibacterial effect is significantly deteriorated.

[0108] Table 1 MIC test results of examples and comparative examples

[0109] MIC(%) Staphylococcus aureus Pseudomonas aeruginosa Escherichia coli Candida albicans Aspergillus niger Example 1 2 0.8 0.8 1.2 1.2 Example 2 1 0.4 0.4 0.6 0.6 Example 3 0.4 0.16 0.16 0.24 0.24 Example 4 2.1 0.9 0.9 1.3 1.4 Example 5 2.2 1 1 1.4 1.3 Example 6 1.8 0.7 0.7 1.1 1.1 Comparative Example 1 2.8 1.1 1.15 1.6 1.7 Comparative Example 2 2.6 1 1.1 1.5 1.6 Comparative Example 3 2.7 1 1.1 1.6 1.7 Comparative Example 4 4 1.6 1.7 2.4 2.4 Comparative Example 5 3 1.1 1.2 1.8 1.8

[0110] Effect Example 2

[0111] In order to explore the mite removal efficacy of the insect repellent, antibacterial and mite removal liposome preparation provided by the present invention, the insect repellent, antibacterial and mite removal liposome preparations of the embodiment and the samples of the comparative example were prepared with deionized water as solvent, and the concentration gradients of 0.1%, 0.5%, 1% and 2% were prepared to test the mite removal rate. The mite removal test method is as follows:

[0112] Take 7 culture dishes, place one in the center of the adhesive board as the central culture dish, and place the remaining 6 culture dishes evenly around the central culture dish in a petal shape, and use transparent tape of the same width to stick the edges between each culture dish (to act as a bridge). Then fix the 7 culture dishes on the adhesive board in the following order: Figure 4 shown.

[0113] Place the samples and control samples in the 6 outer petri dishes at intervals. Add the corresponding amount of sample to the sample, and do not add any substance to the control sample. Spread the sample evenly, flatly and tightly on the bottom of the petri dish, and put 0.05g of mite feed in the center of the sample and control sample. Then put (2000±200) living mites (adult mites and / or nymphs) on the center petri dish.

[0114] Place the sticky plate assembly containing the test mites and feed in a constant temperature and humidity incubator at (25±2)°C and relative humidity (75±5)%. After 24 hours of incubation, observe with a dissecting microscope or stereo microscope and count the number of surviving mites in the sample culture dish and the control sample culture dish using appropriate methods.

[0115] The calculation formula for the mite removal rate is:

[0116] Q=(BT) / B×100%

[0117] Wherein, Q is the mite removal rate (%), B is the average number of surviving mites in three control samples, and T is the average number of surviving mites in three test samples.

[0118] The calculated value of the mite removal rate is taken as the result. When the calculated value is negative, it is expressed as "0%"; when the calculated value is greater than 99%, it is expressed as ">99%". The mite removal rate test results of different samples at different concentrations are shown in Table 2 and Figure 5 shown.

[0119] From Table 2 and Figure 5 It can be seen that:

[0120] (1) The insect repellent, antibacterial and mite removal liposome preparations provided in the embodiments of the present invention all have good mite removal effects. When the mass concentration is 0.5%, a mite removal effect of more than 85% can be achieved after 24 hours of treatment. Among them, the insect repellent, antibacterial and mite removal liposome preparation provided in Example 3 has the best mite removal effect, and a mite removal rate of 99% can be achieved at a concentration of 0.1%.

[0121] (2) By comparing Example 1 with Comparative Examples 1-3, it can be seen that the mite removal rates of the liposome preparation in Example 1 at different concentrations are higher than those of Comparative Examples 1-3, and the mite removal rate advantage is more obvious at lower concentrations. This verifies that forming a liposome formulation and further solidifying the liposomes can significantly improve the mite removal effect and achieve a higher mite removal rate at low concentrations.

[0122] (3) From the comparison between Example 1 and Comparative Examples 4 and 5, it can be seen that the dosage ratio of the efflux pump inhibitor, liposome wall material and liposome curing agent in the present invention also has a significant effect on the comprehensive mite removal effect of all components. When the dosage ratio in Comparative Examples 4 and 5 is outside the scope of the present invention, the mite removal effect of the insect repellent, antibacterial and mite removal liposome preparations is significantly deteriorated.

[0123] Table 2 Test results of mite removal rate of examples and comparative examples

[0124]

[0125]

[0126] Effect Example 3

[0127] In order to explore the anthelmintic efficacy of the anthelmintic, antibacterial and mite-removing liposome preparation provided by the present invention, the anthelmintic, antibacterial and mite-removing liposome preparations of the embodiments and the samples of the comparative examples were prepared with deionized water as solvent, and samples with a mass concentration of 1%, 3%, 5% and 10% were respectively prepared to test the anthelmintic effect:

[0128] Refer to GB / T 13917.9-2009 Indoor efficacy test evaluation of public health insecticides for pesticide registration Part 9: Repellents for implementation.

[0129] The specific test methods are as follows:

[0130] First, do the attack power test: put 300 test insects in the mosquito cage, expose 40mm×40mm skin on the back of the tester's hand, and cover the rest of the body. Put your hand into the mosquito cage and stay there for 2 minutes, and observe closely. If you find a mosquito landing, shake your arm to drive it away before its mouthparts pierce the skin, and it is recorded as one test insect landing. If more than 30 test insects land on the tester and the test insects, the attack power is qualified, and this person and the mosquitoes in this cage can be used for the repellent test.

[0131] After the attack power test is qualified, the avoidance test is conducted: 4 or more testers with qualified attack power are selected (half male and half female, and no alcohol, tea or coffee should be consumed before and during the test, and no products containing fragrances should be used). Draw a 50mm×50mm skin area on the back of each hand. One hand is evenly smeared with the repellent to be tested at a dose of 1.5pL / cm (liquid repellent sample), exposing 40mm×40mm of the skin, and the rest is tightly covered. The other hand is a blank control. Apply the repellent for 2 hours, put the hand into a mosquito cage with qualified attack power for 2 minutes, and observe whether there are mosquitoes coming to land and suck blood. After that, the test is conducted every 1 hour. As long as one mosquito comes to suck blood, the repellent sample is judged to be invalid. Record the effective protection time (h) of the repellent sample. Each time, the control hand is first tested for control, and the test insects with qualified attack power can continue the test. If the test insect's attack power is unqualified, it is necessary to replace the qualified test insect for the test.

[0132] Evaluation of test results:

[0133] The quality of mosquito repellent products is evaluated based on the effective protection time (h) of the repellent on the test personnel. The efficacy results are divided into two levels, A and B. Products that do not meet the B standard are unqualified. Grade A: effective protection time ≥ 6.0h, Grade B: effective protection time ≥ 4.0h. The insect repellent test results are shown in Table 3.

[0134] From Table 3, we can see that:

[0135] (1) The anthelmintic, antibacterial and mite-removing liposome preparations provided in the embodiments of the present invention all have good anthelmintic effects, and can achieve an anthelmintic protection time of at least 4 hours at a concentration of 1%; among them, the anthelmintic, antibacterial and mite-removing liposome preparations provided in Examples 1-3 and 6 have the best effects, and can achieve an anthelmintic protection time of at least 6 hours.

[0136] (2) By comparing the insect repellent effects of Example 1 and Comparative Examples 1-3, it can be seen that the liposome dosage form and liposome solidification treatment of the insect repellent, antibacterial and mite removal liposome preparation provided by the present invention can also significantly improve its insect repellent effect. In Comparative Example 1, no liposome formation and no solidification, in Comparative Example 2, no liposome solidification, and in Comparative Example 3, no liposome formation will all lead to a significant deterioration of the insect repellent effect. This may be related to the fact that liposomes can better act on the skin and enable the composite insect repellent to form a repellent barrier.

[0137] (3) From the comparison between Example 1 and Comparative Examples 4 and 5, it can be seen that when the usage ratio of the efflux pump inhibitor in Comparative Example 4, the liposome wall material and the liposome curing agent in Comparative Example 5 is not appropriate or exceeds the specified range of the present invention, the anthelmintic test will fail and an effective anthelmintic effect cannot be achieved for a long time.

[0138] Table 3 Insect repellent test results of the embodiments and comparative examples

[0139]

[0140]

[0141] Effect Example 4

[0142] In order to explore the sustained-release effect of the anthelmintic, antibacterial and mite-removing liposome preparation provided by the present invention, the anthelmintic, antibacterial and mite-removing liposome preparations of the examples and the samples of the comparative examples were subjected to the following release rate test:

[0143] Weigh 10 g of Example 1, Comparative Example 1 and Comparative Example 2 respectively, add them into 100 g of normal saline (0.9% NaCl sterile aqueous solution), and detect the content of marker-glycyrrhizic acid in the normal saline at 2h, 4h, 8h, 16h, 24h, 32h and 48h after addition. The ratio of the measured content to the theoretical maximum content is the release rate of the active ingredient of Example 1, Comparative Example 1 and Comparative Example 2.

[0144] The release rate results of the active ingredients in Example 1, Comparative Example 1 and Comparative Example 2 are shown in Tables 4 and Figure 6 shown.

[0145] From Table 4 and Figure 6 It can be seen that the sample of comparative example 1 that did not form a liposome dosage form was completely released in about 8 hours; the sample of comparative example 2 that did not undergo the liposome solidification step released slowly, but also reached a high release rate of 89% after 48 hours; and the insect repellent, antibacterial and mite removal liposome preparation of Example 1, the release rate of the active ingredient was less than 50% after 48 hours of treatment with physiological saline. It can be seen from the curve that it can achieve stable and slow release, prevent the explosive release of the active ingredient, and provide safer and longer-lasting application performance; at the same time, this result also indirectly confirms the formation of the liposome dosage form in Example 1 and the successful solidification and stabilization of the liposome morphology.

[0146] Table 4 Release test results of Example 1 and Comparative Examples 1 and 2

[0147] Time / release rate (%) 2h 4h 8h 16h 24h 32h 48h Example 1 10 15 19 25 31 37 45 Comparative Example 1 75 95 99 99 99 99 99 Comparative Example 2 40 48 57 76 82 85 89

[0148] Effect Example 5

[0149] In order to explore the safety of the anthelmintic, antibacterial and mite-removing liposome preparation provided by the present invention, the anthelmintic, antibacterial and mite-removing liposome preparation prepared in the embodiment was subjected to multiple skin irritation tests, and the test concentration was 5% mass concentration:

[0150] This project is carried out in accordance with the "Technical Specifications for Cosmetic Safety" and the testing agency is the Guangzhou Quality Supervision and Inspection Institute.

[0151] The results of multiple skin irritation tests of the example samples at 5% concentration are shown in Table 5. As can be seen from Table 5, at 5% mass concentration, the results of multiple skin irritation tests are all non-irritating, indicating that the insect repellent, antibacterial, and mite removal liposome preparation provided by the present invention is mild to the skin and has high safety.

[0152] Based on the above results, it can be seen that the insect repellent, antibacterial and mite removal liposome preparation provided by the present invention can achieve good antibacterial, mite removal and insect repellent effects at a lower addition concentration, and has a sustained release property, and can provide safer and longer-lasting application performance. At the same time, its ingredients are mild and non-irritating to human skin at higher concentrations, and it has high application value in the field of daily chemicals.

[0153] Table 5 Skin irritation test results of the insect repellent, antibacterial and mite removal liposome preparations of the present invention

[0154] 5%wt sample Example 1 Example 2 Example 3 Example 4 Example 5 Example 6 Irritant Non-irritating Non-irritating Non-irritating Non-irritating Non-irritating Non-irritating

[0155] Application Examples

[0156] The insect repellent, antibacterial and mite removal liposome preparation provided by the present invention can be further used as an additive in laundry detergent and pet shampoo. Taking Example 1 and Example 2 as examples, the specific formulas can be referred to as shown in Tables 6 and 7.

[0157] The insect repellent, antibacterial and mite removal liposome preparation of Example 1 can give the laundry detergent the functions of insect repellent, antibacterial and mite removal, and can be used for daily washing and maintenance of fabrics such as clothing, bed sheets, curtains, etc.;

[0158] The insect repellent, antibacterial and mite removal liposome preparation of Example 2 can give pet shampoo the functions of insect repellent, antibacterial and mite removal, and can be used for daily cleaning and maintenance of hairy pets such as cats, dogs, hamsters, etc.

[0159] Table 6 Application formula of Example 1 in laundry detergent

[0160]

[0161]

[0162] Table 7 Application formula of Example 2 in pet shampoo

[0163]

[0164]

[0165] Finally, it should be noted that the above embodiments are only used to illustrate the technical solution of the present invention rather than to limit the scope of protection of the present invention. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solution of the present invention can be modified or replaced by equivalents without departing from the essence and scope of the technical solution of the present invention.

Claims

1. An insect repellent, antibacterial and mite removing liposome preparation, characterized in that: The invention comprises liposome contents, liposome wall materials, liposome curing agent and water, wherein the liposome contents comprise bactericide, insecticide, antibacterial agent and efflux pump inhibitor; the antibacterial agent is a polyol, and the liposome curing agent is a C16-C22 fatty acid; The insect repellent, antibacterial and mite removing liposome preparation comprises the following components by mass percentage: 0.1-10% bactericide, 0.1-10% insecticide, 1-40% antibacterial agent, 0.1-10% efflux pump inhibitor, 0.1-10% liposome wall material, 0.01-1% liposome curing agent and the balance water.

2. The insect repellent, antibacterial and mite removing liposome preparation according to claim 1, characterized in that: The invention comprises the following components by mass percentage: 0.5-10% of bactericide, 1-8% of insecticide, 20-35% of antibacterial agent, 1-5% of efflux pump inhibitor, 2-3% of liposome wall material, 0.2-0.5% of liposome curing agent and the balance of water.

3. The insect repellent, antibacterial and mite removing liposome preparation according to claim 1, characterized in that: The bactericide is at least one of diclosan, phenoxyethanol, polyhexamethylene hydrochloride, sodium benzoate, potassium sorbate, methylparaben, propylparaben, methylisothiazolinone, methylchloroisothiazolinone, DMDM ​​hydantoin and chlorphenesin.

4. The insect repellent, antibacterial and mite removing liposome preparation according to claim 1, characterized in that: The insecticide is at least one of natural menthanediol, pyrethrin, pepper extract, onion extract, orange peel extract, garlic extract, honeysuckle extract, sophora flavescens extract, ginkgo extract, derris extract, wormwood extract, castor seed extract, and datura extract.

5. The insect repellent, antibacterial and mite removing liposome preparation according to claim 1, characterized in that: The antibacterial agent is at least one of glycerol, ethanol, propylene glycol, butylene glycol, pentanediol and hexylene glycol.

6. The insect repellent, antibacterial and mite removing liposome preparation according to claim 1, characterized in that: The efflux pump inhibitor is at least one of glycyrrhizic acid, methyl pyrrolidone, artesunate, antisense nucleic acid, oligonucleotide and amikacin.

7. The insect repellent, antibacterial and mite removing liposome preparation according to claim 1, characterized in that: The liposome wall material is at least one of lecithin, hydrogenated lecithin, hydroxylated lecithin, hydrogenated lysolecithin, lysolecithin and soybean lecithin.

8. The insect repellent, antibacterial and mite removing liposome preparation according to claim 1, characterized in that: The liposome solidifying agent is at least one of palmitic acid, linoleic acid, linolenic acid, oleic acid, stearic acid, arachidic acid, and behenic acid.

9. The method for preparing the insect repellent, antibacterial and mite removing liposome preparation according to any one of claims 1 to 8, characterized in that: The following steps are involved: (1) dispersing a fungicide, an insecticide and an efflux pump inhibitor in deionized water, heating and keeping the water warm, and mixing the water to obtain a phase A; (2) dissolving the liposome wall material in the antibacterial agent, heating and keeping warm, and mixing to obtain phase B; (3) adding the phase B dropwise into the phase A under heating, insulation and stirring to obtain phase C; The heating and heat preservation temperature is 35-65°C; (4) homogenizing the phase C to obtain phase D, then heating the phase to 45-75° C., dispersing the liposome solidifying agent in the phase D, and cooling the phase to 15-30° C. to obtain the insect repellent, antibacterial, and mite removal liposome preparation.

10. Use of the insect repellent, antibacterial and mite removing liposome preparation according to any one of claims 1 to 8 in the field of daily chemicals.

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