4-hydroxymethylphenylboronic acid modified amphotericin B derivative as well as preparation method and application thereof

Abstract

The invention relates to a 4-hydroxymethylphenylboronic acid modified amphotericin B derivative as well as a preparation method and application thereof, and belongs to the technical field of medicine preparation. Amino groups on trehalose amine in amphotericin B molecules are modified through 4-hydroxymethyl phenylboronic acid, and the amphotericin B derivative with a novel structure is obtained. Compared with amphotericin B, the derivative has the characteristics of low hemolysis toxicity, low kidney cell toxicity and low acute toxicity. In addition, the invention also provides a preparation method of the derivative, and the prepared derivative has better antifungal activity and has a wide market prospect in the aspect of treating systemic deep fungal infection.

Description

Technical Field

[0001] The invention belongs to the technical field of drug preparation, and relates to a 4-hydroxymethylphenylboronic acid modified amphotericin B derivative, a preparation method and an application thereof. Background Art

[0002] Amphotericin B is a polyene antifungal drug with a broad antifungal spectrum and is the drug of choice for the treatment of deep fungal infections. Amphotericin B has strong hemolytic toxicity, acute toxicity and renal cell toxicity, especially the nephrotoxicity problem, which has not been effectively solved so far. A large number of studies have shown that by modifying the structure of amphotericin B molecules, its toxicity can be significantly reduced, but its antibacterial activity will be affected to a certain extent.

[0003] Some researchers have introduced methoxypolyethylene glycol, bisaminoalkyl, sarcosine and other groups into the sugar amine of amphotericin B molecules to prepare a series of derivatives. The toxicity of the obtained derivatives may be lower than that of amphotericin B, but the antibacterial activity is greatly affected or even lost. In addition, the synthesis steps of these derivatives are many, generally more than 3 steps, the preparation method is complicated, and the product yield is low, which is one of the reasons why no new amphotericin B derivatives have been developed so far.

[0004] Phenylboronic acid is the most common triggering group in reactive oxygen species (ROS) reactions. It is easily hydrolyzed under physiological conditions to release the original drug and can be used for the research and development of prodrugs. To this end, we used 4-hydroxymethylphenylboronic acid, 4-hydroxyethylphenylboronic acid, 3-hydroxymethylphenylboronic acid, (4-(2-hydroxyethyl)phenyl)boronic acid, p-carboxylphenylboronic acid and 2-fluoro-5-hydroxymethylphenylboronic acid to modify the sugar amine on the amphotericin B molecule, and finally screened out a 4-hydroxymethylphenylboronic acid modified amphotericin B derivative. Compared with amphotericin B, this derivative has the advantages of less hemolytic effect, low renal cell toxicity and acute toxicity, and has good antifungal activity. It can be degraded to generate amphotericin B under the ROS microenvironment of the fungal infection site to further exert a strong antifungal effect. In addition, the preparation method of this derivative is simple, with fewer synthesis steps and mild synthesis reaction conditions, which is very feasible for product development. Summary of the invention

[0005] In view of this, one of the objects of the present invention is to provide a 4-hydroxymethylphenylboronic acid modified amphotericin B derivative; the second object of the present invention is to provide a method for preparing a 4-hydroxymethylphenylboronic acid modified amphotericin B derivative; the third object of the present invention is to provide an application of a 4-hydroxymethylphenylboronic acid modified amphotericin B derivative in the treatment of fungal infectious diseases; the fourth object of the present invention is to provide a pharmaceutical composition comprising a 4-hydroxymethylphenylboronic acid modified amphotericin B derivative.

[0006] In order to achieve the above object, the present invention provides the following technical solutions:

[0007] 1. A 4-hydroxymethylphenylboronic acid modified amphotericin B derivative, the structural formula of the derivative is shown in the figure below:

[0008]

[0009] 2. The preparation method of the above-mentioned 4-hydroxymethylphenylboronic acid modified amphotericin B derivative, the reaction formula of the method is:

[0010]

[0011] Preferably, the method comprises the following steps:

[0012] (1) Disperse 4-hydroxymethylphenylboronic acid, di(p-nitrobenzene) carbonate and an organic base in acetonitrile, react at 25-45° C. for 1-12 hours under N2 protection, evaporate and remove the solvent to obtain a reactant, add a mixed solution of ethyl acetate: petroleum ether: glacial acetic acid in a volume ratio of 60:40:1 to dissolve the reactant for column chromatography. Use silica gel powder as the stationary phase and a mixed solution of ethyl acetate: petroleum ether: glacial acetic acid in a volume ratio of 60:40:1 as the mobile phase to separate and purify the reactant by column chromatography, collect the eluent, evaporate and remove the solvent, add n-hexane to stir and disperse, filter and collect the product, and vacuum dry to obtain a 4-hydroxymethylphenylboronic acid intermediate;

[0013] (2) Amphotericin B, 4-hydroxymethylphenylboronic acid intermediate and organic base are dissolved in N,N-dimethylformamide and reacted at 20-40°C for 1-6 hours under N2 protection. The reaction solution is poured into methyl tert-butyl ether for precipitation, the product is collected by centrifugation, acetone is added to fully disperse, and then poured into methyl tert-butyl ether for precipitation, the product is collected by centrifugation, and the "dispersion-precipitation" operation is repeated 3 times, the product is collected by filtration, and vacuum dried to obtain a yellow powder of 4-hydroxymethylphenylboronic acid modified amphotericin B derivative.

[0014] Further preferably, the molar ratio of 4-hydroxymethylbenzeneboronic acid, di(p-nitrobenzene) carbonate and organic base in step (1) is 1:2-3:1-2;

[0015] The organic base is triethylamine or N,N-diisopropylethylamine;

[0016] The mass volume ratio of the 4-hydroxymethylphenylboronic acid and acetonitrile is 10:1-3, mg / ml;

[0017] The mass volume ratio of the 4-hydroxymethylphenylboronic acid and ethyl acetate: petroleum ether: glacial acetic acid mixed solution (for dissolution) is 10:0.5-1.5, mg / ml;

[0018] The mass volume ratio of the 4-hydroxymethylphenylboronic acid and n-hexane is 10:1-2, mg / ml.

[0019] Preferably, the molar ratio of amphotericin B, 4-hydroxymethylphenylboronic acid intermediate and organic base in step (2) is 1:1.2-1.6:1.5-3.5;

[0020] The organic base is triethylamine or N,N-diisopropylethylamine;

[0021] The mass volume ratio of amphotericin B and N,N-dimethylformamide is 20:1-3, mg / ml;

[0022] The volume ratio of N,N-dimethylformamide to methyl tert-butyl ether is 1:8-10;

[0023] The mass volume ratio of amphotericin B to acetone is 5:1-2, mg / ml;

[0024] The volume ratio of the acetone to methyl tert-butyl ether is 1:2-3.

[0025] 3. Use of the above-mentioned 4-hydroxymethylphenylboronic acid modified amphotericin B derivative in the preparation of drugs for inhibiting fungal growth or treating fungal infectious diseases.

[0026] 4. A pharmaceutical composition comprising the above-mentioned 4-hydroxymethylphenylboronic acid modified amphotericin B derivative.

[0027] Preferably, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier, and the pharmaceutically acceptable carrier is any one of distearoylphosphatidylethanolamine-polyethylene glycol, soybean lecithin or egg yolk lecithin.

[0028] Preferably, the pharmaceutical composition is in the form of an injection.

[0029] The beneficial effects of the present invention are:

[0030] 1. The present invention discloses a 4-hydroxymethylphenylboronic acid modified amphotericin B derivative. The present invention uses 4-hydroxymethylphenylboronic acid to modify the glucosamine on the amphotericin B molecule. The obtained derivative has the characteristics of low hemolytic effect, low renal cell toxicity, low acute toxicity, and exhibits good antifungal activity, and has good application prospects in the treatment of fungal infectious diseases.

[0031] 2. The present invention also discloses a method for preparing a 4-hydroxymethylphenylboronic acid modified amphotericin B derivative, which is simple, has few synthetic steps, and has mild synthetic reaction conditions. First, the hydroxyl group on 4-hydroxymethylphenylboronic acid is activated by di(p-nitrobenzene) carbonate to obtain an activated ester intermediate, which is then reacted with amphotericin B after purification to obtain a derivative, and its molecular structure design is innovative to a certain extent, providing a new research idea for the design and development of ROS microenvironment-responsive and degradable amphotericin B derivatives.

[0032] Other advantages, objectives and features of the present invention will be described in the following description to some extent, and to some extent, will be obvious to those skilled in the art based on the following examination and study, or can be taught from the practice of the present invention. The objectives and other advantages of the present invention can be realized and obtained through the following description. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] In order to make the purpose, technical solutions and advantages of the present invention more clear, the present invention will be described in detail below in conjunction with the accompanying drawings, wherein:

[0034] Figure 1 is the H NMR spectrum of the 4-hydroxymethylphenylboronic acid modified amphotericin B derivative prepared in Example 1;

[0035] Figure 2 It is the hemolysis rate test result of in vitro hemolysis test;

[0036] Figure 3 This is the result of acute toxicity evaluation in mice;

[0037] Figure 4 The results of Cryptococcal fungal load experiments in the lungs and brains of infected mice;

[0038] Figure 5 The results of the Candida albicans fungal load experiment in the kidneys of infected mice;

[0039] Figure 6 It is the structural formula of the amphotericin B derivative modified with 4-hydroxymethylphenylboronic acid of the present invention. DETAILED DESCRIPTION

[0040] The following describes the embodiments of the present invention by specific examples, and those skilled in the art can easily understand other advantages and effects of the present invention from the contents disclosed in this specification. The present invention can also be implemented or applied through other different specific embodiments, and the details in this specification can also be modified or changed in various ways based on different viewpoints and applications without departing from the spirit of the present invention. It should be noted that the illustrations provided in the following embodiments only illustrate the basic concept of the present invention in a schematic manner, and the following embodiments and features in the embodiments can be combined with each other without conflict.

[0041] Example 1

[0042] A 4-hydroxymethylphenylboronic acid modified amphotericin B derivative is prepared, which specifically comprises the following steps:

[0043] (1) Weigh 100 mg of 4-hydroxymethylphenylboronic acid, di(p-nitrobenzene) carbonate (400 mg, 2 eq.) and triethylamine (100 mg, 1.5 eq.) and disperse them in 12 ml of acetonitrile. React at 40° C. for 8 h under N2 protection. Evaporate to remove the solvent. Add 6 ml of a mixed solution of ethyl acetate: petroleum ether: glacial acetic acid in a volume ratio of 60:40:1 to dissolve completely and prepare for column chromatography. Use silica gel powder as the solid phase and a mixed solution of ethyl acetate: petroleum ether: glacial acetic acid in a volume ratio of 40:60:1 as the mobile phase to separate and purify the reactant by column chromatography. Collect the eluent, evaporate to remove the solvent, add 15 ml of n-hexane to stir and disperse, collect the product by filtration, and dry in vacuo to obtain the 4-hydroxymethylphenylboronic acid intermediate (yield 76%).

[0044] (2) Weigh 50 mg of amphotericin B, 4-hydroxymethylphenylboronic acid intermediate (23 mg, 1.4 eq.) and triethylamine (16 mg, 3 eq.) and dissolve in 3 ml of N,N-dimethylformamide. React at 25°C for 4 h under N2 protection. Pour the reaction solution into 30 ml of methyl tert-butyl ether for precipitation. Collect the product by centrifugation. Add 15 ml of acetone to fully disperse it. Pour it into 40 ml of methyl tert-butyl ether for precipitation. Collect the product by centrifugation. Repeat the "dispersion-precipitation" operation 3 times. Filter and collect the product. Dry in vacuo to obtain a yellow powder of 4-hydroxymethylphenylboronic acid-modified amphotericin B derivative. 1 H NMR (400MHz) spectrum is attached Figure 1 shown.

[0045] Example 2

[0046] The performance test of the 4-hydroxymethylphenylboronic acid modified amphotericin B derivative prepared in Example 1 was performed:

[0047] 1. In vitro hemolysis test

[0048] Amphotericin B and amphotericin B derivative modified with 4-hydroxymethylphenylboronic acid were dissolved in DMSO to prepare a 5mM concentrated solution. The concentrated amphotericin B solution was diluted to a concentration of 5 to 400 μM with a 5% glucose solution. At the same time, the concentrated amphotericin B derivative modified with 4-hydroxymethylphenylboronic acid was diluted to a concentration of 50 to 4000 μM with a 5% glucose solution. A 2ml centrifuge tube was taken, and 500 μl of 2% erythrocyte suspension, 450 μl of normal saline and 50 μl of drug solution were added. The mixture was gently mixed to make the concentration of amphotericin B in the erythrocyte suspension 0.25 to 20 μM, and the concentration of amphotericin B derivative modified with 4-hydroxymethylphenylboronic acid in the erythrocyte suspension 2.5 to 200 μM. Pure water and normal saline were used as positive and negative controls, respectively. The above solution was incubated at 37±0.5℃ for 1h and 3h, the solution was taken out and centrifuged at 2000rpm for 10min, 200μl of the supernatant was taken and the absorbance was measured at 540nm. The hemolysis rate (%) was calculated as follows:

[0049] Hemolysis rate (%) = (Abs 样品 –Abs 阴 ) / (Abs 阳 –Abs 阴 )×100.

[0050] Data fitting was performed based on the calculated hemolysis rate to obtain the EH50 values ​​(the concentration of the drug corresponding to the hemolysis rate reaching 50%) corresponding to amphotericin B and the amphotericin B derivative modified with 4-hydroxymethylphenylboronic acid.

[0051] The results of hemolysis rate determination show that (Appendix Figure 2 ), amphotericin B has a strong hemolytic effect, with an EH50 value of 1.4μM at 1h and 1.1μM at 3h, while the EH50 value of the amphotericin B derivative modified with 4-hydroxymethylphenylboronic acid is greater than 200μM, and its hemolytic toxicity is significantly reduced compared with amphotericin B.

[0052] 2. In vitro pharmacodynamic evaluation

[0053] The minimum inhibitory concentration (MIC) of amphotericin B and 4-hydroxymethylphenylboronic acid modified amphotericin B derivatives against Cryptococcus (Cryptococcus neoformans H99, Cryptococcus gattii R256) and Candida (Candida parapsilosis ATCC 22019, Candida albicans SC 5314, Candida albicans ATCC 90028, Candida auris and Candida krusei ATCC 6258) was determined by microplate dilution method. The specific method is as follows:

[0054] Amphotericin B and 4-hydroxymethylphenylboronic acid modified amphotericin B derivatives were dissolved in DMSO to prepare a 1 mM concentrated solution, and the drug concentrated solution was diluted to a series of concentrations of 0.3125-40 μM with 5% glucose solution. 160 μl of bacterial solution (1.25×10 3 CFU / ml) and 40μl of different concentrations of drug solution, so that the concentration of the drug in the well plate is 0.0625-8μM. Two groups of controls are set up, one group mixes 160μl of bacterial solution with 40μl of normal saline, and the other group mixes 160μl of culture medium with 40μl of normal saline. The 96-well plate is placed in a constant temperature incubator at 30±0.5℃, and the OD value at 490nm is measured after 48h and the MIC is calculated. The MIC value is defined as the minimum concentration that completely inhibits fungal growth.

[0055] The MIC determination results are shown in Table 1. Compared with amphotericin B, although the in vitro antifungal activity of the 4-hydroxymethylphenylboronic acid modified amphotericin B derivative was reduced (the MIC value was twice that of amphotericin B), it still showed good in vitro antifungal activity.

[0056] Table 1 Evaluation of in vitro antifungal activity of amphotericin B and its derivatives

[0057]

[0058] 3. Evaluation of renal cell toxicity

[0059] The toxicity of amphotericin B and amphotericin B derivatives modified with 4-hydroxymethylphenylboronic acid to human renal cortical proximal tubule epithelial cells HK-2 was investigated. The specific method is as follows: amphotericin B was dissolved in DMSO to prepare a 5mM concentrated solution, and the amphotericin B concentrated solution was diluted to a series of concentrations of 5 to 160μM with a 5% glucose solution. Amphotericin B derivative modified with 4-hydroxymethylphenylboronic acid was dissolved in DMSO to prepare a 50mM concentrated solution, and the amphotericin B concentrated solution was diluted to a series of concentrations of 10 to 1280μM with a 5% glucose solution. HK-2 cells in the logarithmic growth phase were digested and inoculated into a sterile 96-well plate, 160μl per well, and placed in a cell culture incubator at 37±0.5℃ for 24h. 40 μl of drug solution of different concentrations was added to each well, so that the concentration of amphotericin B in the cell suspension was 1-32 μM, and the concentration of amphotericin B derivative modified with 4-hydroxymethylphenylboronic acid in the cell suspension was 2-256 μM, and the cells were cultured in a cell culture incubator at 37±0.5°C for 48 h. The culture medium was removed, and the cells were washed with PBS 2-3 times, and then 180 μl of new culture medium and 20 μl of MTT solution (5 mg / ml) were added. After incubation at 37±0.5°C for 4 h, the culture medium was discarded, 150 μl of DMSO was added, and the cells were shaken for 15 min. The cells were placed in an ELISA reader and the OD value was measured at 490 nm to calculate the cell survival rate.

[0060] The cell viability calculation formula is as follows:

[0061] Cell survival rate (%) = (OD 样品 -OD 空白 ) / (OD 对照 -OD 空白 )×100

[0062] The experimental results showed that amphotericin B has strong renal cell toxicity, killing more than 90% of HK-2 cells at a concentration of 64 μM, while the 4-hydroxymethylphenylboronic acid modified amphotericin B derivative did not show obvious renal cell toxicity at a concentration of up to 256 μM. Compared with amphotericin B, the renal cell toxicity of amphotericin B derivatives was significantly reduced.

[0063] 4. Acute toxicity test

[0064] Healthy ICR mice were randomly divided into groups, with 10 mice in each group. Egg yolk lecithin was used as the carrier material, and amphotericin B and amphotericin B derivatives modified with 4-hydroxymethylphenylboronic acid were prepared into liposome preparations, which were injected into the tail vein of mice. The symptoms of mice in each group and the death within 14 days were observed and recorded, and the mouse survival curve was drawn (see Appendix). Figure 3). When the amphotericin B liposome solution was administered at a dose of 5 mg / kg, all mice died immediately, showing symptoms of poisoning such as convulsions, breathing difficulties, and incontinence. In contrast, when the amphotericin B derivative liposome solution modified with 4-hydroxymethylphenylboronic acid was administered at a dose of 60 mg / kg, all mice behaved normally and all survived within 14 days. This shows that the acute toxicity of the amphotericin B derivative modified with 4-hydroxymethylphenylboronic acid is significantly reduced compared to amphotericin B.

[0065] In addition, healthy KM mice were randomly divided into groups, with 10 mice in each group. Using distearoylphosphatidylethanolamine-polyethylene glycol 2000 as the carrier material, amphotericin B and 4-hydroxymethylphenylboronic acid modified amphotericin B derivatives were prepared into micelle preparations, which were injected into the tail vein of mice, and the symptoms of mice in each group and the death within 14 days were observed and recorded. When amphotericin B micelle solution was administered at a dose of 5 mg / kg, all mice died within 30 minutes, accompanied by toxic reactions such as convulsions, spasms, incontinence, and rapid breathing. In contrast, when amphotericin B derivative micelle solution modified with 4-hydroxymethylphenylboronic acid was administered at a dose of 100 mg / kg, some mice showed varying degrees of mental depression within 30 minutes, but quickly returned to normal, and no mice died within two weeks.

[0066] 5. In vivo pharmacodynamic evaluation in mice

[0067] Cryptococcal fungal load experiment in mice: Cryptococcus neoformans H99 was injected into the tail vein to establish a mouse lung and brain fungal infection model. The specific method is as follows:

[0068] (1) Absorb H99 Cryptococcus culture (1×10 7 CFU / ml), and the model was established by injecting bacterial solution into the tail vein of ICR mice (dose 100μl / 20g). 24h after infection, 4-hydroxymethylphenylboronic acid modified amphotericin B derivative liposome solution and control commercial preparation AmBisome were injected into the tail vein once a day for 3 consecutive times, with a dose of 2mg / kg. 24h after the third administration, the mice were dissected on a sterile operating table, the lungs and brain tissues were removed, washed with sterilized saline, and dried with filter paper. After adding saline according to the weight of lung or brain tissue and saline mass volume ratio of 1:3 (g / ml), the homogenate was diluted, and 50μl was applied to YPD solid culture medium, placed in a 30℃ incubator for 48h, and the fungal growth in each culture plate was observed, and the colony count was performed.

[0069] From the attached Figure 4It can be seen that compared with the untreated model group, the 4-hydroxymethylphenylboronic acid-modified amphotericin B derivative liposomes have obvious antifungal activity, significantly reducing the number of fungi in the lungs and brain of mice, and there is no significant difference in the therapeutic effect with the commercially available preparation AmBisome.

[0070] In vivo Candida albicans fungal load experiment in mice: The mouse kidney fungal infection model was established by tail vein injection of Candida albicans ATCC 90028. The entire experimental method was the same as the Cryptococcal fungal load experiment. The results are shown in the attached Figure 5 Compared with the untreated model group, the 4-hydroxymethylphenylboronic acid-modified amphotericin B derivative liposomes had significant antifungal activity and significantly reduced the number of fungi in the kidneys of mice, and there was no significant difference in the therapeutic effect with the commercially available preparation AmBisome.

[0071] The above results indicate that the 4-hydroxymethylphenylboronic acid modified amphotericin B derivative prepared by the present invention has good in vivo antifungal activity.

[0072] In summary, the present invention uses 4-hydroxymethylphenylboronic acid to modify the amino group on the trehalose amine in the amphotericin B molecule to obtain a novel 4-hydroxymethylphenylboronic acid modified amphotericin B derivative (whose structural formula is shown in the attached figure). Figure 6 As shown in the figure, compared with amphotericin B, it has the characteristics of less hemolytic effect, lower nephrotoxicity and lower acute toxicity. In addition, the present invention also provides a method for preparing the derivative, and the prepared derivative has good in vitro and in vivo antifungal activity and has broad market prospects in the treatment of systemic deep fungal infections.

[0073] 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 it. 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 purpose and scope of the technical solution, which should be included in the scope of the claims of the present invention.

Claims

1. A 4-hydroxymethylphenylboronic acid modified amphotericin B derivative, characterized in that: The structural formula of the derivative is shown in the figure below:

2. The method for preparing the 4-hydroxymethylphenylboronic acid modified amphotericin B derivative according to claim 1, characterized in that: The reaction formula of the method is:

3. The preparation method according to claim 2, characterized in that: The method comprises the following steps: (1) Disperse 4-hydroxymethylphenylboronic acid, di(p-nitrobenzene) carbonate and an organic base in acetonitrile, react at 25-45° C. for 1-12 hours under N2 protection, evaporate and remove the solvent to obtain a reactant, add a mixed solution of ethyl acetate: petroleum ether: glacial acetic acid in a volume ratio of 60:40:1 to dissolve the reactant for column chromatography. Use silica gel powder as the stationary phase and a mixed solution of ethyl acetate: petroleum ether: glacial acetic acid in a volume ratio of 60:40:1 as the mobile phase to separate and purify the reactant by column chromatography, collect the eluent, evaporate and remove the solvent, add n-hexane to stir and disperse, filter and collect the product, and vacuum dry to obtain a 4-hydroxymethylphenylboronic acid intermediate; (2) Amphotericin B, 4-hydroxymethylphenylboronic acid intermediate and organic base are dissolved in N,N-dimethylformamide and reacted at 20-40°C for 1-6 hours under N2 protection. The reaction solution is poured into methyl tert-butyl ether for precipitation, the product is collected by centrifugation, acetone is added to fully disperse, and then poured into methyl tert-butyl ether for precipitation, the product is collected by centrifugation, and the "dispersion-precipitation" operation is repeated 3 times, the product is collected by filtration, and vacuum dried to obtain a yellow powder of 4-hydroxymethylphenylboronic acid modified amphotericin B derivative.

4. The preparation method according to claim 3, characterized in that: The molar ratio of 4-hydroxymethylphenylboronic acid, di(p-nitrobenzene) carbonate and organic base in step (1) is 1:2-3:1-2, the organic base is triethylamine or N,N-diisopropylethylamine, the mass volume ratio of 4-hydroxymethylphenylboronic acid and acetonitrile is 10:1-3, mg / ml, the mass volume ratio of 4-hydroxymethylphenylboronic acid and ethyl acetate: petroleum ether: glacial acetic acid mixed solution (for dissolution) is 10:0.5-1.5, mg / ml, and the mass volume ratio of 4-hydroxymethylphenylboronic acid and n-hexane is 10:1-2, mg / ml; In step (2), the molar ratio of amphotericin B, 4-hydroxymethylphenylboronic acid intermediate and organic base is 1:1.2-1.6:1.5-3.5, the organic base is triethylamine or N,N-diisopropylethylamine, the mass volume ratio of amphotericin B and N,N-dimethylformamide is 20:1-3, mg / ml, the volume ratio of N,N-dimethylformamide and methyl tert-butyl ether is 1:8-10, the mass volume ratio of amphotericin B and acetone is 5:1-2, mg / ml, and the volume ratio of acetone to methyl tert-butyl ether is 1:2-3.

5. Use of the 4-hydroxymethylphenylboronic acid modified amphotericin B derivative according to claim 1 in the preparation of a medicament for inhibiting fungal growth or treating fungal infectious diseases.

6. A pharmaceutical composition, characterized in that Amphotericin B derivative modified with 4-hydroxymethylphenylboronic acid as claimed in claim 1.

7. The composition according to claim 6, characterized in that The pharmaceutical composition further comprises a pharmaceutically acceptable carrier, and the pharmaceutically acceptable carrier is any one of distearoyl phosphatidylethanolamine-polyethylene glycol, soybean lecithin or egg yolk lecithin.

8. The composition according to claim 6, characterized in that The dosage form of the pharmaceutical composition is injection.

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