High-performance voriconazole liposome as well as preparation method and application thereof

Abstract

The invention discloses a high-performance voriconazole liposome as well as a preparation method and application thereof, and the high-performance voriconazole liposome is prepared from the following components in parts by mass: 30 to 100 parts of phospholipid, 0 to 30 parts of cholesterol and 2.5 to 10 parts of voriconazole. According to the voriconazole liposome preparation disclosed by the invention, voriconazole is wrapped in phospholipid double layers, so that the solubility of voriconazole is increased, and meanwhile, a medicine is protected inside the liposome, so that the stability of the medicine is improved. And moreover, the encapsulation efficiency and the release rate of the liposome are improved by changing the types of phospholipids and the ratio of the phospholipids to the cholesterol. The preparation method of the voriconazole liposome is simple, and the preparation temperature is high, so that the residual organic reagent in the rotary evaporation process is effectively removed.

Description

Technical Field

[0001] The invention belongs to the technical field of poorly soluble drug preparations, and particularly relates to a voriconazole liposome preparation and a preparation method and application thereof. Background Art

[0002] Fungal infections are diseases caused by fungi that can affect the skin, mucous membranes, internal organs, and even the entire body. Depending on the site and severity of infection, fungal infections can be divided into superficial infections and deep (invasive) infections. Superficial fungal infections mainly invade the skin, hair, and nails, while deep (invasive) fungal infections mainly invade deep tissues or internal organs and are common in people with weakened immune systems.

[0003] Voriconazole is a broad-spectrum triazole antifungal drug that inhibits lanosterol 14α-demethylase (CYP51) in the fungal cell membrane and prevents the synthesis of ergosterol, thereby destroying the integrity of the fungal cell membrane and causing fungal cell death. It is mainly used to treat invasive fungal infections such as aspergillosis, candidiasis, and other infections caused by rare fungi. However, voriconazole has extremely low solubility in water and its chemical properties are unstable in solution.

[0004] Liposome is a microcapsule structure composed of a phospholipid bilayer membrane. It is composed of amphiphilic phospholipid molecules (such as lecithin and soybean lecithin), with the hydrophilic head facing the water phase and the hydrophobic tail facing the inside, forming a bilayer structure with a hydrophilic core and a hydrophobic bilayer membrane, which can encapsulate water-soluble and fat-soluble drugs. According to the structure and size of liposomes, they are mainly divided into unilamellar liposomes, multilamellar liposomes and multivesicular liposomes. The advantages of liposomes are mainly that they can improve drug stability, increase solubility, targeted delivery, reduce adverse reactions and achieve sustained release effects. Summary of the invention

[0005] The purpose of the present invention is to provide a high-performance voriconazole liposome and a preparation method and application thereof, which has high encapsulation rate and release rate, increases the solubility of the drug and improves the stability of the drug.

[0006] To achieve the above object, the present invention adopts the following technical solutions:

[0007] In a first aspect, the present invention provides a high-performance voriconazole liposome.

[0008] A high-performance voriconazole liposome consists of the following components in parts by weight: 30 to 100 parts of phospholipids, 0 to 30 parts of cholesterol, and 2.5 to 10 parts of voriconazole.

[0009] The cholesterol content may be 0 or not.

[0010] In some embodiments, the high-performance voriconazole liposomes are composed of 30 to 100 parts by mass of phospholipids and 2.5 to 10 parts by mass of voriconazole, and do not contain cholesterol.

[0011] In some embodiments, the high-performance voriconazole liposomes are composed of 30 to 100 parts by mass of phospholipids, 0 to 30 parts by mass of cholesterol, and 2.5 to 10 parts by mass of voriconazole, and the cholesterol content is not zero.

[0012] In some embodiments, the voriconazole liposomes can be unilamellar liposomes, multilamellar liposomes or multivesicular liposomes.

[0013] In some embodiments, the voriconazole liposome is a structure having a hydrophilic inner core and a hydrophobic bilayer membrane, formed by voriconazole being encapsulated by a phospholipid bilayer membrane.

[0014] Preferably, the phospholipid is one or more of soybean lecithin, DSPC, DPPC, and DMPC.

[0015] Preferably, the voriconazole liposomes are composed of the following components: 30-100 mg phospholipid content, 0-30 mg cholesterol content, and 2.5-10 mg voriconazole.

[0016] Preferably, the encapsulation rate of the voriconazole liposome is ≥40%, and the release rate is ≥70%.

[0017] In a preferred embodiment of the present invention, the composition of the voriconazole liposome is: 60 mg DMPC, 0 mg cholesterol and 10 mg voriconazole. The encapsulation efficiency of the voriconazole liposome of this composition is 91.32%.

[0018] In a preferred embodiment of the present invention, the composition of the voriconazole liposome is: 50 mg DPPC, 10 mg cholesterol and 5 mg voriconazole. The release rate of the voriconazole liposome of this composition is 99.04%.

[0019] In a second aspect, the present invention provides a method for preparing high-performance voriconazole liposomes.

[0020] A method for preparing high-performance voriconazole liposomes comprises the following steps:

[0021] (1) dissolving voriconazole, phospholipid and cholesterol in an organic solvent to obtain solution A, or dissolving voriconazole and phospholipid in an organic solvent to obtain solution A;

[0022] (2) placing solution A in a rotary evaporator to remove the organic solvent until a lipid film is formed;

[0023] (3) Add phosphate buffer to the lipid film obtained in step (2) and hydrate to obtain liposome solution B.

[0024] Preferably, in step (1), the organic solvent is chloroform.

[0025] Preferably, in step (2), the rotary evaporation temperature is 60°C.

[0026] Preferably, in step (2), the rotation speed of the rotary evaporator is 100 rpm / min.

[0027] Preferably, in step (3), the pH of the phosphate buffer is 7.4.

[0028] Preferably, in step (3), the hydration temperature is 60°C.

[0029] Preferably, in step (3), the hydration time is 1 h.

[0030] Preferably, in step (3), hydration is carried out by rotary evaporation.

[0031] In a third aspect, the present invention provides the use of the voriconazole liposomes.

[0032] The application of the high-performance voriconazole liposome in the preparation of antifungal drugs.

[0033] Preferably, the antifungal drug is used to treat invasive fungal infections.

[0034] Preferably, the invasive fungal infection is aspergillosis, candidiasis and other infections caused by rare fungi.

[0035] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0036] Unless otherwise indicated, the term "DSPC" refers to distearoylphosphatidylcholine, the term "DPPC" refers to dipalmitoylphosphatidylcholine, and the term "DMPC" refers to dimyristoylphosphatidylcholine.

[0037] Beneficial effects: Compared with the prior art, the present invention has the following advantages: the voriconazole liposome preparation of the present invention encapsulates voriconazole in a phospholipid bilayer, thereby increasing the solubility of voriconazole and protecting the drug inside the liposome, thereby improving the stability of the drug. Furthermore, by changing the type of phospholipids and the ratio of phospholipids to cholesterol, the encapsulation rate and release rate of the liposome are increased. The preparation method of the voriconazole liposome is simple, and the preparation temperature is relatively high, so that the residual organic reagents in the rotary evaporation process are effectively removed. BRIEF DESCRIPTION OF THE DRAWINGS

[0038] Figure 1 is the encapsulation efficiency of liposomes with DSPC as phospholipid under different raw material ratios;

[0039] Figure 2 is the encapsulation efficiency of liposomes with DPPC as phospholipid under different raw material ratios;

[0040] Figure 3 is the encapsulation efficiency of liposomes with DMPC as phospholipid under different raw material ratios;

[0041] Figure 4 is the encapsulation efficiency of liposomes with lecithin as the phospholipid at different raw material ratios;

[0042] Figure 5 The release rate of liposomes with DSPC as phospholipid under different raw material ratios;

[0043] Figure 6 is the release rate of liposomes with DPPC as phospholipid under different raw material ratios;

[0044] Figure 7 The release rate of liposomes with DMPC as phospholipid under different raw material ratios;

[0045] Figure 8 The release rate of liposomes whose phospholipid is lecithin under different raw material ratios. DETAILED DESCRIPTION

[0046] The technical solution of the present disclosure is further described below in conjunction with specific embodiments. However, the following embodiments are merely examples of the present disclosure and do not represent or limit the scope of protection of the present disclosure. In the following embodiments, unless otherwise specified, the reagents and consumables used are purchased from ordinary suppliers in the field, and the experimental methods and technical means used are conventional methods and means in the field.

[0047] Example 1

[0048] The voriconazole liposome preparation and preparation method thereof of the present embodiment are as follows:

[0049] Liposome composition:

[0050] Voriconazole 10 mg

[0051] DSPC 30mg

[0052] Cholesterol 30mg

[0053] The preparation method comprises: dissolving voriconazole, DSPC and cholesterol in 10 mL of chloroform, and stirring evenly to obtain solution A. The solution A is placed in a rotary evaporator at a speed of 100 rpm / min, and the organic solvent is removed by rotary evaporation at 60° C. until a lipid film is formed. Then, 10 mL of phosphate buffer is added to the lipid film, and the rotary evaporation is continued at 60° C. for 1 hour, so that the lipid film is gradually hydrated into liposome solution B.

[0054] Example 2

[0055] The voriconazole liposome preparation and preparation method thereof of the present embodiment are as follows:

[0056] Liposome composition:

[0057] Voriconazole 10 mg

[0058] DSPC 60mg

[0059] Cholesterol 0mg

[0060] The preparation method comprises: dissolving voriconazole and DSPC in 10 mL of chloroform, and stirring evenly to obtain solution A. The solution A is placed in a rotary evaporator at a speed of 100 rpm / min, and the organic solvent is removed by rotary evaporation at 60° C. until a lipid film is formed. Then, 10 mL of phosphate buffer is added to the lipid film, and the rotary evaporation is continued at 60° C. for 1 hour, so that the lipid film is gradually hydrated into liposome solution B.

[0061] Example 3

[0062] The voriconazole liposome preparation and preparation method thereof of the present embodiment are as follows:

[0063] Liposome composition:

[0064] Voriconazole 10 mg

[0065] DSPC 100mg

[0066] Cholesterol 0mg

[0067] The preparation method comprises: dissolving voriconazole and DSPC in 10 mL of chloroform, and stirring evenly to obtain solution A. The solution A is placed in a rotary evaporator at a speed of 100 rpm / min, and the organic solvent is removed by rotary evaporation at 60° C. until a lipid film is formed. Then, 10 mL of phosphate buffer is added to the lipid film, and the rotary evaporation is continued at 60° C. for 1 hour, so that the lipid film is gradually hydrated into liposome solution B.

[0068] Example 4

[0069] The voriconazole liposome preparation and preparation method thereof of the present embodiment are as follows:

[0070] Liposome composition:

[0071] Voriconazole 10 mg

[0072] DPPC 30mg

[0073] Cholesterol 30mg

[0074] The preparation method comprises: dissolving voriconazole, DPPC and cholesterol in 10 mL of chloroform, and stirring evenly to obtain solution A. The solution A is placed in a rotary evaporator at a speed of 100 rpm / min, and the organic solvent is removed by rotary evaporation at 60° C. until a lipid film is formed. Then, 10 mL of phosphate buffer is added to the lipid film, and the rotary evaporation is continued at 60° C. for 1 hour, so that the lipid film is gradually hydrated into liposome solution B.

[0075] Example 5

[0076] The voriconazole liposome preparation and preparation method thereof of the present embodiment are as follows:

[0077] Liposome composition:

[0078] Voriconazole 10 mg

[0079] DPPC 60mg

[0080] Cholesterol 0mg

[0081] The preparation method comprises: dissolving voriconazole and DPPC in 10 mL of chloroform, and stirring evenly to obtain solution A. The solution A is placed in a rotary evaporator at a speed of 100 rpm / min, and the organic solvent is removed by rotary evaporation at 60° C. until a lipid film is formed. Then, 10 mL of phosphate buffer is added to the lipid film, and the rotary evaporation is continued at 60° C. for 1 hour, so that the lipid film is gradually hydrated into liposome solution B.

[0082] Example 6

[0083] The voriconazole liposome preparation and preparation method thereof of the present embodiment are as follows:

[0084] Liposome composition:

[0085] Voriconazole 5 mg

[0086] DPPC 50mg

[0087] Cholesterol 10mg

[0088] The preparation method comprises: dissolving voriconazole, DPPC and cholesterol in 10 mL of chloroform, and stirring evenly to obtain solution A. The solution A is placed in a rotary evaporator at a speed of 100 rpm / min, and the organic solvent is removed by rotary evaporation at 60° C. until a lipid film is formed. Then, 10 mL of phosphate buffer is added to the lipid film, and the rotary evaporation is continued at 60° C. for 1 hour, so that the lipid film is gradually hydrated into liposome solution B.

[0089] Example 7

[0090] The voriconazole liposome preparation and preparation method thereof of the present embodiment are as follows:

[0091] Liposome composition:

[0092] Voriconazole 10 mg

[0093] DMPC 30mg

[0094] Cholesterol 30mg

[0095] The preparation method comprises: dissolving voriconazole, DMPC and cholesterol in 10 mL of chloroform, and stirring evenly to obtain solution A. The solution A is placed in a rotary evaporator at a speed of 100 rpm / min, and the organic solvent is removed by rotary evaporation at 60° C. until a lipid film is formed. Then, 10 mL of phosphate buffer is added to the lipid film, and the rotary evaporation is continued at 60° C. for 1 hour, so that the lipid film is gradually hydrated into a liposome solution B.

[0096] Example 8

[0097] The voriconazole liposome preparation and preparation method thereof of the present embodiment are as follows:

[0098] Liposome composition:

[0099] Voriconazole 10 mg

[0100] DMPC 50mg

[0101] Cholesterol 10mg

[0102] The preparation method comprises: dissolving voriconazole, DMPC and cholesterol in 10 mL of chloroform, and stirring evenly to obtain solution A. The solution A is placed in a rotary evaporator at a speed of 100 rpm / min, and the organic solvent is removed by rotary evaporation at 60° C. until a lipid film is formed. Then, 10 mL of phosphate buffer is added to the lipid film, and the rotary evaporation is continued at 60° C. for 1 hour, so that the lipid film is gradually hydrated into a liposome solution B.

[0103] Example 9

[0104] The voriconazole liposome preparation and preparation method thereof of the present embodiment are as follows:

[0105] Liposome composition:

[0106] Voriconazole 10 mg

[0107] DMPC 60mg

[0108] Cholesterol 0mg

[0109] The preparation method comprises: dissolving voriconazole and DMPC in 10 mL of chloroform, and stirring evenly to obtain solution A. The solution A is placed in a rotary evaporator at a speed of 100 rpm / min, and the organic solvent is removed by rotary evaporation at 60° C. until a lipid film is formed. Then, 10 mL of phosphate buffer is added to the lipid film, and the rotary evaporation is continued at 60° C. for 1 hour, so that the lipid film is gradually hydrated into liposome solution B.

[0110] Example 10

[0111] The voriconazole liposome preparation and preparation method thereof of the present embodiment are as follows:

[0112] Liposome composition:

[0113] Voriconazole 10 mg

[0114] DMPC 100mg

[0115] Cholesterol 10mg

[0116] The preparation method comprises: dissolving voriconazole, DMPC and cholesterol in 10 mL of chloroform, and stirring evenly to obtain solution A. The solution A is placed in a rotary evaporator at a speed of 100 rpm / min, and the organic solvent is removed by rotary evaporation at 60° C. until a lipid film is formed. Then, 10 mL of phosphate buffer is added to the lipid film, and the rotary evaporation is continued at 60° C. for 1 hour, so that the lipid film is gradually hydrated into a liposome solution B.

[0117] Embodiment 11

[0118] The voriconazole liposome preparation and preparation method thereof of the present embodiment are as follows:

[0119] Liposome composition:

[0120] Voriconazole 10 mg

[0121] Soy lecithin 45mg

[0122] Cholesterol 15mg

[0123] The preparation method comprises: dissolving voriconazole, soybean lecithin and cholesterol in 10 mL of chloroform, and stirring evenly to obtain solution A. The solution A is placed in a rotary evaporator at a speed of 100 rpm / min, and the organic solvent is removed by rotary evaporation at 60° C. until a lipid film is formed. Then, 10 mL of phosphate buffer is added to the lipid film, and the rotary evaporation is continued at 60° C. for 1 hour, so that the lipid film is gradually hydrated into a liposome solution B.

[0124] Example 12

[0125] The voriconazole liposome preparation and preparation method thereof of the present embodiment are as follows:

[0126] Liposome composition:

[0127] Voriconazole 5 mg

[0128] Soy lecithin 50mg

[0129] Cholesterol 10mg

[0130] The preparation method comprises: dissolving voriconazole, soybean lecithin and cholesterol in 10 mL of chloroform, and stirring evenly to obtain solution A. The solution A is placed in a rotary evaporator at a speed of 100 rpm / min, and the organic solvent is removed by rotary evaporation at 60° C. until a lipid film is formed. Then, 10 mL of phosphate buffer is added to the lipid film, and the rotary evaporation is continued at 60° C. for 1 hour, so that the lipid film is gradually hydrated into a liposome solution B.

[0131] Example 13

[0132] The voriconazole liposome preparation and preparation method thereof of the present embodiment are as follows:

[0133] Liposome composition:

[0134] Voriconazole 2.5 mg

[0135] Soy lecithin 50mg

[0136] Cholesterol 10mg

[0137] The preparation method comprises: dissolving voriconazole, soybean lecithin and cholesterol in 10 mL of chloroform, and stirring evenly to obtain solution A. The solution A is placed in a rotary evaporator at a speed of 100 rpm / min, and the organic solvent is removed by rotary evaporation at 60° C. until a lipid film is formed. Then, 10 mL of phosphate buffer is added to the lipid film, and the rotary evaporation is continued at 60° C. for 1 hour, so that the lipid film is gradually hydrated into a liposome solution B.

[0138] Test Example 1

[0139] The encapsulation efficiency of liposomes was determined by ultrafiltration centrifugation.

[0140] A certain amount of liposome solution B in Examples 1 to 13 was placed in an ultrafiltration centrifuge tube and centrifuged at 10000 rpm / min for 10 min to obtain a filtrate. The content of free voriconazole in the filtrate was determined by an ultraviolet spectrophotometer, and then the liposome encapsulation efficiency was determined.

[0141] Figures 1 to 4 It can be seen that the encapsulation efficiency of the liposome is above 40%, with the highest being 91.32%, and the corresponding raw material composition is: 60 mg DMPC, 0 mg cholesterol and 10 mg voriconazole.

[0142] Test Example 2

[0143] The cumulative release rate of liposomes was determined by dialysis method.

[0144] A certain amount of liposome solution B in Examples 1 to 13 was placed in a dialysis bag, the dialysis medium was a pH 7.4 phosphate buffer, and then placed in a constant temperature water bath at 37°C, with a stirring speed of 400 rpm / min, and samples were taken at intervals, and the same volume of fresh medium was added for fluid replenishment. The voriconazole content in the dialysis medium was then determined by an ultraviolet spectrophotometer, and the release rate of the liposomes was then determined.

[0145] Figures 5 to 8 It can be seen that the liposomes are basically released completely in about 120 minutes, and the release rate is above 70%, with the highest being 99.04%. The corresponding raw material composition is 50 mg DPPC, 10 mg cholesterol and 5 mg voriconazole.

[0146] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A high-performance voriconazole liposome, characterized in that: The invention is composed of the following components in parts by mass: 30 to 100 parts of phospholipids, 0 to 30 parts of cholesterol, and 2.5 to 10 parts of voriconazole.

2. A high-performance voriconazole liposome according to claim 1, characterized in that: The voriconazole liposome is a structure with a hydrophilic core and a hydrophobic double-layer membrane formed by wrapping voriconazole with a phospholipid double-layer membrane.

3. The high-performance voriconazole liposome according to claim 1, characterized in that: The phospholipids are one or more of soybean lecithin, DSPC, DPPC and DMPC.

4. The high-performance voriconazole liposome according to claim 1, characterized in that: The voriconazole liposomes are composed of the following components: 30-100 mg phospholipid content, 0-30 mg cholesterol content, and 2.5-10 mg voriconazole.

5. The high-performance voriconazole liposome according to claim 1, characterized in that: The voriconazole liposome has an encapsulation rate of ≥40% and a release rate of ≥70%.

6. A method for preparing the high-performance voriconazole liposomes according to claim 1, characterized in that: The following steps are involved: (1) dissolving voriconazole, phospholipid and cholesterol in an organic solvent to obtain solution A, or dissolving voriconazole and phospholipid in an organic solvent to obtain solution A; (2) placing solution A in a rotary evaporator to remove the organic solvent until a lipid film is formed; (3) Add phosphate buffer to the lipid film obtained in step (2) and hydrate to obtain liposome solution B.

7. The method for preparing high-performance voriconazole liposomes according to claim 6, characterized in that: In the step (1), the organic solvent is chloroform.

8. The method for preparing high-performance voriconazole liposomes according to claim 6, characterized in that: In the step (2), the temperature of the rotary evaporation is 60° C. and the speed of the rotary evaporator is 100 rpm / min.

9. The method for preparing high-performance voriconazole liposomes according to claim 6, characterized in that: In the step (3), the pH of the phosphate buffer is 7.4, the hydration temperature is 60° C., and the hydration time is 1 hour.

10. Use of the high-performance voriconazole liposome according to claim 1 in the preparation of antifungal drugs.

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