Nano preparation as well as preparation method and application thereof

Abstract

The invention relates to the technical field of nano materials, and discloses a nano preparation as well as a preparation method and application thereof. The nano preparation contains a phospholipid compound, an inhibitor A and an inhibitor B, the inhibitor A is selected from at least one of rapamycin, fluconazole, amphotericin B and caspofungin, and the inhibitor B is selected from at least one of deferasirox, deferoxamine, deferoxone, dimercaptosuccinic acid and penicillamine. The nano preparation has a good bacteriostatic and bactericidal effect on biphasic fungi, has a good treatment effect on systemic infection, and can weaken the toxic and side effects of drugs.

Description

Technical Field

[0001] The present invention relates to the technical field of nanomaterials, and in particular to a nano preparation, a preparation method and an application thereof. Background Art

[0002] Dimorphic fungal infections, particularly Candida albicans infections, remain rampant worldwide, resulting in a mortality rate exceeding 60%. These fungi are characterized by their presence in both yeast and hyphal forms. The yeast form primarily spreads through the bloodstream, while the hyphal form facilitates tissue invasion and releases numerous toxins. Current treatments for fungal infections generally target all yeast forms while neglecting to inhibit the hyphal form. Furthermore, most antibiotics have toxic side effects.

[0003] Nanocarrier drug delivery systems can be prepared from various materials (such as lipids, metals, and inorganic materials), and can achieve transport across biological membrane barriers through multiple mechanisms, optimizing the delivery of drugs to sites of infection and other lesions, while minimizing non-targeted drug distribution and increasing the selectivity of drug delivery.

[0004] Although most of the drugs loaded in nanoformulations are first-line clinical drugs, their antibacterial effects on dimorphic fungi still need to be improved. Summary of the Invention

[0005] The purpose of the present invention is to overcome the problem that the antibacterial effect of drugs on diphasic fungi in the prior art needs to be improved, and to provide a nano preparation and its preparation method and application. The nano preparation has good antibacterial and bactericidal effects on diphasic fungi, good therapeutic effects on systemic infections, and can reduce the toxic and side effects of drugs.

[0006] In order to achieve the above objectives, the first aspect of the present invention provides a nanoformulation, which contains a phospholipid compound, an inhibitor A and an inhibitor B, wherein the inhibitor A is selected from at least one of rapamycin, fluconazole, amphotericin B and caspofungin, and the inhibitor B is selected from at least one of deferasirox, deferoxamine, deferiprone, dimercaptosuccinic acid and penicillamine.

[0007] Preferably, in the nanoformulation, the mass ratio of the inhibitor A to the inhibitor B is 1:1-50.

[0008] Further preferably, in the nanoformulation, the content of the inhibitor A is 0.01-0.08 wt%, more preferably 0.01-0.04 wt%; the content of the inhibitor B is 0.1-0.5 wt%, more preferably 0.1-0.4 wt%.

[0009] More preferably, in the nanoformulation, the content of the phospholipid compound is 2.8-5.5 wt %.

[0010] Preferably, the inhibitor B is dimercaptosuccinic acid.

[0011] Preferably, the inhibitor A is rapamycin.

[0012] Preferably, the phospholipid compound is egg yolk lecithin.

[0013] Preferably, the average particle size of the nanoformulation is 35-50 nm.

[0014] A second aspect of the present invention provides a method for preparing a nanoformulation, the method comprising: mixing a phospholipid compound, an inhibitor A and an inhibitor B and then homogenizing the mixture; The inhibitor A is selected from at least one of rapamycin, fluconazole, amphotericin B and caspofungin, and the inhibitor B is selected from at least one of deferasirox, deferoxamine, deferiprone, dimercaptosuccinic acid and penicillamine.

[0015] Preferably, the mass ratio of the inhibitor A to the inhibitor B is 1:1-40.

[0016] Further preferably, the mass ratio of the inhibitor A to the inhibitor B is 1:2-20.

[0017] Further preferably, the mass ratio of the phospholipid compound to the inhibitor A is 60-550:1.

[0018] More preferably, the mass ratio of the phospholipid compound to the inhibitor A is 150-300:1.

[0019] Preferably, the inhibitor B is dimercaptosuccinic acid.

[0020] Preferably, the inhibitor A is rapamycin.

[0021] More preferably, the phospholipid compound is egg yolk lecithin.

[0022] Preferably, the homogenization process is ultrasonic homogenization or high-pressure homogenization; Preferably, the ultrasonic homogenization conditions include at least: ultrasonic power of 200-400W, ultrasonic time of 3-6min; The conditions for the high-pressure homogenization include at least: a pressure of 500-700 Bar and a number of 5-15 times.

[0023] The third aspect of the present invention provides a nanoformulation prepared by the preparation method described in the second aspect.

[0024] The fourth aspect of the present invention provides a use of the nanoformulation described in the first or third aspect in the preparation of a drug for resisting dimorphic fungal infection.

[0025] Preferably, the diphasic fungal infection is Candida albicans.

[0026] Through the above technical solution, the nanoformulation provided by the present invention, inhibitor A is selected from at least one of rapamycin, fluconazole, amphotericin B and caspofungin, and inhibitor B is selected from at least one of deferasirox, deferoxamine, deferiprone, dimercaptosuccinic acid and penicillamine. Through the interaction between the phospholipid compound, inhibitor A and inhibitor B, the growth of fungi in yeast form and hyphal form can be simultaneously inhibited, and better antibacterial and bactericidal effects are achieved on biphasic fungi, ultimately achieving better bioavailability and systemic infection treatment effects, and reducing the toxic and side effects of the drug. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 is a particle size distribution diagram of the nanoformulation prepared in Example 10; Figure 2 is a transmission electron micrograph of the nanoformulation prepared in Example 10; Figure 3 is the survival rate of yeast in the nanoformulation prepared in Example 1 and the blank control group; Figure 4 The survival rate of infected mice treated with the nanoformulation prepared in Example 1 and the survival rate of infected mice in the blank control group; Figure 5 Statistical graph of mycelial inhibition rate of the nanoformulation prepared in Example 10 and the blank control group. DETAILED DESCRIPTION

[0028] The endpoints of the ranges and any values ​​disclosed herein are not limited to the precise ranges or values, and these ranges or values ​​should be understood to include values ​​close to these ranges or values. For numerical ranges, the endpoints of each range, the endpoints of each range and individual point values, and the individual point values ​​can be combined with each other to obtain one or more new numerical ranges, which should be considered to be specifically disclosed herein.

[0029] As described above, the first aspect of the present invention provides a nanoformulation, which contains a phospholipid compound, an inhibitor A and an inhibitor B, wherein the inhibitor A is selected from at least one of rapamycin, fluconazole, amphotericin B and caspofungin, and the inhibitor B is selected from at least one of deferasirox, deferoxamine, deferiprone, dimercaptosuccinic acid and penicillamine.

[0030] During the research process, the inventors found that by limiting inhibitor A to at least one selected from rapamycin, fluconazole, amphotericin B and caspofungin, and limiting inhibitor B to at least one selected from deferasirox, deferoxamine, deferiprone, dimercaptosuccinic acid and penicillamine, the growth of fungi in yeast and hyphal forms can be simultaneously inhibited through the interaction between phospholipid compounds, inhibitor A and inhibitor B, thereby having better antibacterial and bactericidal effects on biphasic fungi, ultimately achieving better bioavailability and systemic infection treatment effects, and reducing the toxic side effects of the drug.

[0031] Preferably, in the nanoformulation, the mass ratio of the inhibitor A to the inhibitor B is 1:1-50, and can be 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:15, 1:20, 1:25, 1:30, 1:35, 1:40, 1:45, 1:50, or any value within a range formed by any two of these ratios. Studies have found that the above mass ratios of inhibitor A and inhibitor B have a better synergistic effect, thereby further inhibiting the growth of yeast and hyphal fungi, and improving the antibacterial and bactericidal effects against dimorphic fungi. In order to further enhance the antibacterial and bactericidal effects on dimorphic fungi, it is further preferred that, in the nanoformulation, the content of the inhibitor A is 0.01-0.08 wt%, and may be 0.01 wt%, 0.02 wt%, 0.03 wt%, 0.04 wt%, 0.05 wt%, 0.06 wt%, 0.07 wt%, 0.08 wt%, or any value within a range formed by any two of these values; and the content of the inhibitor B is 0.1-0.5 wt%, and may be 0.1 wt%, 0.2 wt%, 0.3 wt%, 0.4 wt%, 0.5 wt%, or any value within a range formed by any two of these values. More preferably, in the nanoformulation, the content of the inhibitor A is 0.01-0.04 wt%, and the content of the inhibitor B is 0.1-0.4 wt%.

[0032] Preferably, in the nanoformulation, the content of the phospholipid compound is 2.8-5.5 wt %. Controlling the content of the phospholipid compound within the above range can further improve the antibacterial and bactericidal effects of the nanoformulation on dimorphic fungi.

[0033] Preferably, the inhibitor B is dithiothioate. Studies have shown that dithiothioate interacts effectively with inhibitor A and phospholipid compounds, further enhancing the antibacterial and bactericidal effects of the nanoformulation against diphasic fungi. Furthermore, to further enhance the antibacterial and bactericidal effects of the nanoformulation against diphasic fungi, the antibacterial agent A is preferably rapamycin.

[0034] Preferably, the phospholipid compound is egg yolk lecithin. Egg yolk lecithin has a good interaction effect with dimercaptosuccinic acid and rapamycin, thereby further improving the antibacterial and bactericidal effects of the nanoformulation on dimorphic fungi.

[0035] In order to further improve the antibacterial and bactericidal effects of the nanoformulation on dimorphic fungi, preferably, the average particle size of the nanoformulation is 35-50 nm.

[0036] The second aspect of the present invention provides a method for preparing a nanoformulation, comprising mixing a phospholipid compound, an inhibitor A and an inhibitor B and then homogenizing the mixture; The inhibitor A is selected from at least one of rapamycin, fluconazole, amphotericin B and caspofungin, and the inhibitor B is selected from at least one of deferasirox, deferoxamine, deferiprone, dimercaptosuccinic acid and penicillamine.

[0037] During the research process, the inventors found that by limiting inhibitor A to at least one selected from rapamycin, fluconazole, amphotericin B and caspofungin, and limiting inhibitor B to at least one selected from deferasirox, deferoxamine, deferiprone, dimercaptosuccinic acid and penicillamine, and then mixing the phospholipid compound, inhibitor A and inhibitor B and performing ultrasonic treatment, the prepared nanoformulation can simultaneously inhibit the growth of fungi in yeast and hyphal forms, have better antibacterial and bactericidal effects on biphasic fungi, ultimately achieving better bioavailability and systemic infection treatment effects, and reducing the toxic side effects of the drug.

[0038] According to the present invention, phospholipid compounds, rapamycin, fluconazole, amphotericin B, caspofungin, deferasirox, deferoxamine, deferiprone, dimercaptosuccinic acid and penicillamine can be purchased commercially.

[0039] Preferably, the mass ratio of the inhibitor A to the inhibitor B is 1:1-40, and can be 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:15, 1:20, 1:25, 1:30, 1:35, 1:40, or any value within a range formed by any two of these ratios. Studies have found that controlling the mass ratio of inhibitor A to inhibitor B within the above range can further enhance the interaction between inhibitor A, inhibitor B, and the phospholipid compound, thereby further enhancing the antibacterial and bactericidal effects of the nanoformulation on diphasic fungi. In order to further enhance the antibacterial and bactericidal effects of the nanoformulation on diphasic fungi, it is further preferred that the mass ratio of the inhibitor A to the inhibitor B is 1:2-20.

[0040] Preferably, the mass ratio of the phospholipid compound to the inhibitor A is 60-550:1, and can be 60:1, 100:1, 150:1, 200:1, 250:1, 300:1, 350:1, 400:1, 450:1, 500:1, 550:1, or any value within a range formed by any two of these ratios. Studies have found that controlling the mass ratio of the phospholipid compound to the inhibitor A within the above range can further enhance the interaction between inhibitor A, inhibitor B, and the phospholipid compound, thereby further enhancing the antibacterial and bactericidal effects of the nanoformulation on diphasic fungi. In order to further enhance the antibacterial and bactericidal effects of the nanoformulation on diphasic fungi, it is further preferred that the mass ratio of the phospholipid compound to the inhibitor A is 150-300:1.

[0041] Preferably, the inhibitor B is dimercaptosuccinic acid. Studies have shown that dimercaptosuccinic acid interacts better with inhibitor A and phospholipid compounds, further enhancing the antibacterial and bactericidal effects of the nanoformulation against diphasic fungi. Furthermore, preferably, the antibacterial agent A is rapamycin, as this can further enhance the antibacterial and bactericidal effects of the nanoformulation against diphasic fungi.

[0042] Preferably, the phospholipid compound is egg yolk lecithin. Egg yolk lecithin has a good interaction effect with dimercaptosuccinic acid and rapamycin, thereby further improving the antibacterial and bactericidal effects of the nanoformulation on dimorphic fungi.

[0043] Preferably, the homogenization treatment is ultrasonic homogenization or high-pressure homogenization, and the nanoemulsion can be obtained by the above-mentioned ultrasonic homogenization or high-pressure homogenization. Preferably, the conditions of the ultrasonic homogenization include at least: the ultrasonic power is 200-400W, which can be 200W, 250W, 300W, 350W, 400W, or any value in the range formed by any two of these values; the ultrasonic time is 3-6min, which can be 3min, 4min, 5min, 6min, or any value in the range formed by any two of these values. The conditions of the high-pressure homogenization include at least: the pressure is 500-700Bar, which can be 500W, 550W, 600W, 650W, 700W, or any value in the range formed by any two of these values; the number of times is 5-15 times, which can be 5 times, 7 times, 9 times, 11 times, 13 times, 15 times, or any value in the range formed by any two of these values.

[0044] Preferably, the method further comprises dialyzing the liquid obtained by the ultrasonic treatment to remove free small molecules. The dialysis treatment can be performed using a dialysis bag or an ultrafiltration cup. Further preferably, the dialysis bag used for the dialysis is 40-60 kD and the dialysis time is 15-22 hours, or an ultrafiltration cup with a particle size of 180-220 nm is used.

[0045] Preferably, the solvent may be water and / or ethanol.

[0046] Preferably, the mixing includes: mixing a phospholipid compound and water to obtain an aqueous solution containing the phospholipid compound; mixing inhibitor A and inhibitor B with ethanol respectively to obtain an ethanol solution containing inhibitor A and an ethanol solution containing inhibitor B; and then mixing the aqueous solution containing the phospholipid compound, the ethanol solution containing inhibitor A, and the ethanol solution containing inhibitor B.

[0047] Preferably, when the phospholipid compound and water are mixed, they can be heated to improve the dissolution effect of the phospholipid compound. There is no special requirement for the heating temperature, and the conventional temperature for dissolving phospholipid compounds in the art can be used. There is no special requirement for the heating time, and it is only necessary to completely dissolve the phospholipid compound.

[0048] Preferably, in the aqueous solution containing phospholipid compounds, the concentration of phospholipid compounds is 60-100 mg / mL; in the ethanol solution containing inhibitor A, the solubility of inhibitor A is 1.6-8 mg / mL; in the ethanol solution containing inhibitor B, the solubility of inhibitor B is 6-10 mg / mL.

[0049] A third aspect of the present invention provides a nanoformulation prepared by the above-mentioned preparation method. This nanoformulation can simultaneously inhibit the growth of both yeast and hyphal fungi, exhibiting enhanced antibacterial and bactericidal effects against biphasic fungi, ultimately achieving improved bioavailability and systemic infection treatment efficacy while reducing drug toxic side effects.

[0050] The present invention further provides a method for preparing a drug for treating diphasic fungal infections. The nanoformulation provided by the present invention has better bactericidal and antibacterial effects on diphasic fungi and can be used to prepare a drug for treating diphasic fungal infections.

[0051] Preferably, the diphasic fungal infection is caused by Candida albicans. The nanoformulation provided by the present invention has a better therapeutic effect on Candida albicans.

[0052] According to a particularly preferred embodiment of the present invention, there is provided a method for preparing a nanoformulation, comprising: (1) mixing a phospholipid compound and water and heating the mixture to completely dissolve the phospholipid compound to obtain an aqueous solution containing the phospholipid compound; mixing inhibitor A and ethanol to obtain an ethanol solution containing inhibitor A; mixing inhibitor B and ethanol to obtain an ethanol solution containing inhibitor B; (2) An aqueous solution containing phospholipid compounds, an ethanol solution containing inhibitor A, and an ethanol solution containing inhibitor B are mixed and homogenized by ultrasonication at 200-400W for 3-6 minutes, and the resulting liquid is dialyzed using a 40-60KD dialysis bag for 16-22 hours; The inhibitor A is selected from at least one of rapamycin, fluconazole, amphotericin B and caspofungin, and the inhibitor B is selected from at least one of deferasirox, deferoxamine, deferiprone, dimercaptosuccinic acid and penicillamine. The mass ratio of the inhibitor A, the inhibitor B and the phospholipid compound is 1:1-40:60-550.

[0053] The nanoformulation prepared by the above preparation method can simultaneously inhibit the growth of yeast-type and hyphal-type fungi through the interaction between phospholipid compounds, inhibitor A and inhibitor B, and has better antibacterial and bactericidal effects on biphasic fungi, ultimately achieving better bioavailability and systemic infection treatment effects, and reducing the toxic side effects of the drug.

[0054] The present invention will be described in detail below through examples. In the following examples, egg yolk lecithin was purchased from Sinopharm Group.

[0055] Example 1 240 mg of egg yolk lecithin was dissolved in 3 mL of pure water with heating and stirring. 1 mL of an 8 mg / mL solution of deferasirox in ethanol and 0.5 mL of a 1.6 mg / mL solution of rapamycin in ethanol were added. The mixture was ultrasonically disrupted at 300 W for 4 minutes. Free drug was removed using a dialysis bag with a molecular weight cutoff of 50 kD, resulting in a uniform, precipitate-free nanoformulation. The encapsulated drug concentration was determined by high-performance liquid chromatography. Based on the total drug concentration, the encapsulation efficiency (%) was calculated to be 62% using the formula: (drug concentration after dialysis / drug concentration at administration) × 100%. The average particle size of the particles was 40 nm, as measured by a Malvern particle size analyzer.

[0056] Example 2 A nanoformulation was prepared according to the method of Example 1, except that deferasirox was replaced with deferoxamine. The encapsulation efficiency of the prepared nanoformulation was 64% and the average particle size was 40 nm.

[0057] Example 3 A nanoformulation was prepared according to the method of Example 1, except that deferasirox was replaced by deferiprone. The encapsulation efficiency of the prepared nanoformulation was 57% and the average particle size was 39 nm.

[0058] Example 4 A nanoformulation was prepared according to the method of Example 1, except that the 1.6 mg / mL rapamycin ethanol solution was replaced with an 8 mg / mL fluconazole ethanol solution. The encapsulation efficiency of the prepared nanoformulation was 64% and the average particle size was 40 nm.

[0059] Example 5 A nanoformulation was prepared according to the method of Example 1, except that the 1.6 mg / mL rapamycin ethanol solution was replaced with a 3 mg / mL amphotericin B ethanol solution. The encapsulation efficiency of the prepared nanoformulation was 59% and the average particle size was 38 nm.

[0060] Example 6 A nanoformulation was prepared according to the method of Example 1, except that the 1.6 mg / mL rapamycin ethanol solution was replaced with an 8 mg / mL caspofungin ethanol solution. The encapsulation efficiency of the prepared nanoformulation was 59% and the average particle size was 40 nm.

[0061] Example 7 A nanoformulation was prepared according to the method of Example 4, except that deferasirox was replaced with dimercaptosuccinic acid. The encapsulation efficiency of the prepared nanoformulation was 60% and the average particle size was 45 nm.

[0062] Example 8 A nanoformulation was prepared according to the method of Example 4, except that deferasirox was replaced by deferoxamine, and fluconazole was replaced by amphotericin B. The encapsulation efficiency of the prepared nanoformulation was 60% and the average particle size was 40 nm.

[0063] Example 9 The nanoformulation was prepared according to the method of Example 1, except that penicillamine was used instead of deferasirox. The encapsulation efficiency of the prepared nanoformulation was 55% and the average particle size was 47 nm.

[0064] Example 10 A nanoformulation was prepared according to the method of Example 7, except that fluconazole was replaced by rapamycin. The encapsulation efficiency of the prepared nanoformulation was 58% and the average particle size was 45 nm.

[0065] The particle size distribution of the nanoformulation prepared in this example is shown in FIG. Figure 1 As shown in the transmission electron microscope image Figure 2 As shown. Figure 1 It can be seen that the average particle size of the dimercaptosuccinic acid & rapamycin nanoparticles is about 45nm. Figure 2 It can be seen that the particles of the nanoparticle preparation are round. Example 11 A nanoformulation was prepared according to the method of Example 7, except that fluconazole was replaced by amphotericin B. The encapsulation efficiency of the prepared nanoformulation was 58% and the average particle size was 42 nm.

[0066] Example 12 A nanoformulation was prepared according to the method of Example 7, except that fluconazole was replaced by caspofungin. The encapsulation efficiency of the prepared nanoformulation was 60% and the average particle size was 40 nm.

[0067] Example 13 A nanoformulation was prepared according to the method of Example 5, except that the concentration of the amphotericin B ethanol solution was 4 mg / mL. The encapsulation efficiency of the prepared nanoformulation was 56% and the average particle size was 41 nm.

[0068] Example 14 A nanoformulation was prepared according to the method of Example 1, except that the concentration of the deferasirox ethanol solution was 10 mg / mL. The encapsulation efficiency of the prepared nanoformulation was 56% and the average particle size was 41 nm.

[0069] Example 15 The nanoformulation was prepared according to the method of Example 1, except that the concentration of the deferasirox ethanol solution was 4 mg / mL, the amount of egg yolk lecithin was 220 mg, and the concentration of the rapamycin ethanol solution was 0.8 mg / mL.

[0070] Example 16 The nanoformulation was prepared according to the method of Example 1, except that the amount of the deferasirox ethanol solution was 2 mL, the amount of the egg yolk lecithin was 260 mg, and the amount of the rapamycin ethanol solution was 1 mL.

[0071] Example 17 The nanoformulation was prepared according to the method of Example 16, except that the amount of the deferasirox ethanol solution used was 4 mL.

[0072] Example 18 The nanoformulation was prepared according to the method of Example 1, except that the amount of the deferasirox ethanol solution used was 8 mL.

[0073] Example 19 The nanoformulation was prepared according to the method of Example 1, except that the amount of rapamycin ethanol solution used was 2 mL.

[0074] Example 20 The nanoformulation was prepared according to the method of Example 1, except that the amount of rapamycin ethanol solution used was 4 mL.

[0075] Comparative Example 1 A nanoformulation was prepared according to the method of Example 1, except that deferasirox was replaced with amphotericin B. The encapsulation efficiency of the prepared nanoformulation was 58% and the average particle size was 40 nm.

[0076] Comparative Example 2 A nanoformulation was prepared according to the method of Example 1, except that rapamycin was replaced by deferoxamine. The encapsulation efficiency of the prepared nanoformulation was 60% and the average particle size was 40 nm.

[0077] Comparative Example 3 A nanoformulation was prepared according to the method of Example 1, except that rapamycin was replaced by micafungin. The encapsulation efficiency of the prepared nanoformulation was 61% and the average particle size was 38 nm.

[0078] Comparative Example 4 A nanoformulation was prepared according to the method of Example 1, except that deferasirox was replaced by edetate. The encapsulation efficiency of the prepared nanoformulation was 63% and the average particle size was 39 nm.

[0079] Test Example 1 The physicochemical properties of the nanoformulations are shown in Table 1.

[0080] The content of inhibitor A and inhibitor B in the formulations was determined by high-performance liquid chromatography (Shimadzu, LC40), and the particle encapsulation efficiency was calculated from this. Particle size was measured using a Malvern particle size analyzer. A standard minimum inhibitory concentration (MIC) assay was performed, measuring the yeast inhibitory effect of the formulations 12 hours after administration using a UV spectrophotometer (Shanghai Youke Instrument Co., Ltd., L3). Various particles were administered in a liquid culture environment at 37°C at 1640°C. Samples were then taken for microscopic observation and the inhibition rate of mycelial growth was quantified.

[0081] The inhibition of yeast growth by the drug was determined using the MIC standard test. The nanoformulation obtained in Example 1 was added to the yeast suspension at a dose of 64 ng / ml of rapamycin. The survival rate of the yeast was as follows: Figure 3 As shown, Figure 3 This shows that the nano preparation provided by the present invention has a good inhibitory effect on yeast.

[0082] A mouse infection model was established by injecting Candida albicans into the tail vein. Six hours after the model was established, the mice were divided into a blank control group and an experimental group. The nanoformulation prepared in Example 1 was intravenously injected into the experimental group mice, and the survival of the mice was finally observed. Figure 4 As shown, after treatment with the nanoformulation prepared in Example 1, the survival time of infected mice was greatly improved.

[0083] Under hyphae induction conditions (cultured at 37°C, 1640 culture medium), 12 hours after administration, the cell wall of Candida albicans was stained with calcofluor white, and the morphological characteristics of the fungus were observed under a fluorescence microscope, and the hyphae formation was counted, such as Figure 5 As shown. Figure 5 It can be seen that the inhibition rate of the nanoformulation in Example 10 on mycelium can reach more than 99%.

[0084] Table 1 Physicochemical properties of nanoformulations

[0085] As can be seen from Table 1, the inhibition rate of the embodiment on mycelium is much better than that of comparative example 1 and comparative example 4. The inhibition rate of the embodiment on mycelium and the inhibition rate of Candida albicans yeast are both higher than those of comparative examples 2 and 3, indicating that the nanoformulation provided by the present invention has a good inhibitory effect on both mycelium and Candida albicans yeast.

[0086] The preferred embodiments of the present invention have been described in detail above, but the present invention is not limited thereto. Within the technical concept of the present invention, various simple variations of the technical solution of the present invention may be made, including combining the various technical features in any other appropriate manner. These simple variations and combinations should also be regarded as disclosed in the present invention and fall within the scope of protection of the present invention.

Claims

1. A nano preparation, characterized in that: The nanoformulation contains a phospholipid compound, an inhibitor A and an inhibitor B, wherein the inhibitor A is selected from at least one of rapamycin, fluconazole, amphotericin B and caspofungin, and the inhibitor B is selected from at least one of deferasirox, deferoxamine, deferiprone, dimercaptosuccinic acid and penicillamine.

2. The nanoformulation according to claim 1, characterized in that In the nanoformulation, the mass ratio of the inhibitor A to the inhibitor B is 1:1-50; Preferably, in the nanoformulation, the content of the inhibitor A is 0.01-0.08 wt%, more preferably 0.01-0.04 wt%; the content of the inhibitor B is 0.1-0.5 wt%, more preferably 0.1-0.4 wt%.

3. The nanoformulation according to claim 2, characterized in that In the nanoformulation, the content of the phospholipid compound is 2.8-5.5 wt %.

4. The nanoformulation according to any one of claims 1 to 3, characterized in that The inhibitor B is dimercaptosuccinic acid; Preferably, the inhibitor A is rapamycin; Preferably, the phospholipid compound is egg yolk lecithin; Preferably, the average particle size of the nanoformulation is 35-50 nm.

5. A method for preparing a nano preparation, characterized in that: The method comprises mixing a phospholipid compound, an inhibitor A and an inhibitor B and then performing a homogenization treatment; The inhibitor A is selected from at least one of rapamycin, fluconazole, amphotericin B and caspofungin, and the inhibitor B is selected from at least one of deferasirox, deferoxamine, deferiprone, dimercaptosuccinic acid and penicillamine.

6. The preparation method according to claim 5, characterized in that The mass ratio of the inhibitor A to the inhibitor B is 1:1-40, preferably 1:2-20; Preferably, the mass ratio of the phospholipid compound to the inhibitor A is 60-550:1, more preferably 150-300:

1.

7. The preparation method according to claim 5 or 6, characterized in that: The inhibitor B is dimercaptosuccinic acid; Preferably, the inhibitor A is rapamycin; Preferably, the phospholipid compound is egg yolk lecithin.

8. The preparation method according to claim 5 or 6, characterized in that: The homogenization process is ultrasonic homogenization or high-pressure homogenization; Preferably, the ultrasonic homogenization conditions include at least: ultrasonic power of 200-400W, ultrasonic time of 3-6min; The conditions for the high-pressure homogenization include at least: a pressure of 500-700 Bar and a number of times of 5-15 times.

9. The nanoformulation prepared by the preparation method according to any one of claims 5 to 8.

10. Use of the nanoformulation according to any one of claims 1 to 4 and 9 in the preparation of a drug for treating diphasic fungal infections; Preferably, the diphasic fungal infection is Candida albicans.

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