Pharmaceutical composition containing albumin-bound ertuglifazone nanoparticles and method for preparing the same

Abstract

The present application provides a pharmaceutical composition comprising stable albumin-bound olprinone nanoparticles and a corresponding preparation method. The pharmaceutical composition comprises an active ingredient, olprinone, human blood albumin, a pharmaceutically acceptable buffer salt, and optionally a lyophilized excipient. The present application also provides a lyophilized preparation of albumin-bound olprinone nanoparticles, which can improve the stability, safety and effectiveness of olprinone.

Description

Technical Field

[0001] This invention belongs to the pharmaceutical field and relates to a pharmaceutical composition containing albumin-bound utidelon nanoparticles, its formulation, and the corresponding preparation method. Background Technology

[0002] Utidelon is an epothilone derivative, a macrolide compound, and a secondary metabolite produced by the genetically modified myxobacterium *Cyclocarya pallida*. Studies have shown that epothilone antibiotics exert their antitumor effects by inhibiting the depolymerization of tubulin. The chemical name of Utidelon is (4S,7R,8S,9S,16S)-4,8-dihydroxy-5,5,7,9,13-pentamethyl-16-[(1E)-1-methyl-2-(2-methyl-thiazolyl-4-yl)-vinyl]-hexadecoxetane-13Z-en-2,6-one lactone, and its chemical structure is shown below:

[0003]

[0004] Because utidelon is readily soluble in ethanol, methanol, ethyl acetate, and chloroform, but poorly soluble in water (its saturated solubility in water is less than 1 μg / ml), the marketed utidelon injection (trade name: Utidelon) is not widely available. TM The prescription contains organic solvents such as ethanol and surfactants such as polyoxyethylene castor oil, which may trigger severe allergic reactions. Anti-allergy treatment is required before clinical use to prevent allergic reactions, which reduces the compliance of this type of drug in clinical use, increases adverse reactions in patients, and limits its clinical applicability.

[0005] Patent application CN 104434808A briefly describes a method for preparing albumin-bound nanoparticles with epothilone B as the active ingredient, but it does not elaborate on key quality parameters such as the stability of the prepared epothilone B albumin nanoparticles. Furthermore, in patent application CN104434808A, the drug loading of the epothilone B albumin nanoparticles (based on the weight ratio of epothilone B to albumin) is low, approximately 5%. Since albumin easily forms albumin polymers and other impurities during high-pressure homogenization, a low drug loading will significantly increase the total amount of albumin polymers in the final product, which is detrimental to product quality control.

[0006] Although utidelon belongs to the epothilone class of derivatives, its chemical structure differs significantly from that of epothilone B. This alteration in the drug's chemical structure significantly changes its binding affinity to albumin, resulting in substantial changes to the physical, chemical stability, and physiological characteristics of the prepared albumin-bound drug. This makes the successful development of utidelon into an albumin-bound formulation unpredictable. During their research on the preparation process of albumin-bound utidelon nanoparticles, the inventors discovered that the albumin-bound utidelon nanoparticles prepared using the method described in patent application CN 104434808A exhibited poor particle size stability after homogenization, failing to meet the requirements for clinical application and industrial production.

[0007] Therefore, there is an urgent need for stable albumin-bound utidelon nanoparticles with clinical application prospects and industrial production capability, as well as their formulations and corresponding preparation methods. Summary of the Invention

[0008] To address the aforementioned issues, the inventors unexpectedly discovered during their research that homogenized albumin-bound utidelon nanoparticles were unstable, with their particle size gradually increasing over time. However, by rapidly removing the organic solvent through simultaneous homogenization and solvent removal, the particle size stability of the nanoparticles significantly increased. Furthermore, compared to diluting albumin solutions with water for injection, nanoparticles obtained by homogenizing albumin solutions with phosphate buffer to form an aqueous phase solution exhibited superior stability compared to those prepared with water for injection. Further research revealed that nanoparticles prepared using organic acid buffers such as citrate buffer, malate buffer, and fumarate buffer showed better particle size stability than those prepared with phosphate buffer. Therefore, this invention provides an albumin-bound nanoparticle formulation with utidelon as the main active ingredient. This formulation possesses good physical and chemical stability, improves drug compliance and safety, and has promising clinical application prospects and is suitable for industrial production.

[0009] On one hand, the present invention provides a pharmaceutical composition comprising albumin-bound utedelone nanoparticles with suitable stability, said nanoparticles comprising the active ingredient utedelone, human serum albumin, and a pharmaceutically acceptable buffer salt.

[0010] In some embodiments, the albumin-bound utidelon nanoparticles comprise the active ingredient utidelon and the main carrier material human serum albumin, with the active ingredient utidelon encapsulated in or adsorbed onto the carrier material.

[0011] In some embodiments, in the albumin-bound utedelone nanoparticles, the weight ratio of the active ingredient utedelone to the carrier material human serum albumin is in the range of 0.01:1 to 1:1, for example, a weight ratio of 0.01:1, 0.05:1, 0.1:1, 0.2:1, 0.25:1, 0.28:1, 0.3:1, 1:3, 0.35:1, 0.4:1, 4:9, 0.45:1, 0.5:1, 0.6:1, 2:3, 0.7:1, 0.8:1, or 0.9:1.

[0012] In some embodiments, the albumin-bound utidelon nanoparticles further comprise a pharmaceutically acceptable buffer salt, which may be selected from inorganic acid salts and organic acid salts. In one embodiment, the pharmaceutically acceptable buffer salt is selected from inorganic acid salts, including but not limited to phosphates. In one embodiment, the pharmaceutically acceptable buffer salt is selected from organic acid salts, including but not limited to one or more of citrate, tartrate, acetate, fumarate, ethylenediaminetetraacetic acid (or edetate) salt, and malate. In one embodiment, the pharmaceutically acceptable buffer salt is selected from one or more of phosphates, citrate, tartrate, acetate, fumarate, ethylenediaminetetraacetic acid (or edetate) salt, and malate. In some specific embodiments, the pharmaceutically acceptable buffer salt is selected from phosphates. In some specific embodiments, the pharmaceutically acceptable buffer salt is selected from one or more of citrate, malate, tartrate, fumarate, and ethylenediaminetetraacetic acid. In some specific embodiments, the pharmaceutically acceptable buffer salt is selected from citrate, tartrate, and ethylenediaminetetraacetic acid. In some specific embodiments, the pharmaceutically acceptable buffer salt is selected from citrate.

[0013] The pharmaceutically acceptable buffer salts described in this invention provide better stability for albumin-bound utidelon nanoparticles. The applicant has found that the particle size and product quality of the albumin-bound utidelon nanoparticles described in this invention are independent of the pH value of the buffer salt, but are related to the type of buffer salt. Albumin-bound utidelon nanoparticles containing organic acid salts exhibit better stability than those containing inorganic acid salts.

[0014] In some embodiments, the albumin-bound utidelon nanoparticles further comprise one or more of sucrose, mannitol, glucose, and trehalose.

[0015] In some embodiments, the average particle size of the albumin-bound utidylcholine nanoparticles is in the range of 30 to 200 nm, for example, it can be 30, 40, 50, 60, 70, 80, 90, 100, 105, 108, 110, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 155, 160, 165, 170, 175, 180, 185, 190, 195 or 200 nm, or a range between any two of the above values. Methods for measuring average particle size include, but are not limited to, sedimentation, sieving, microscopic observation, and laser particle size distribution.

[0016] In some embodiments, the pharmaceutical composition is in lyophilized powder form. In some embodiments, the lyophilized powder form of the pharmaceutical composition does not include lyophilized excipients. In some embodiments, the lyophilized powder form of the pharmaceutical composition includes pharmaceutically acceptable lyophilized excipients. The applicant has found that the type of lyophilized excipient does not significantly affect the particle size and stability of the albumin-bound utedelone nanoparticles described in this invention. Therefore, it is contemplated that the type of lyophilized excipient is not particularly limited. In some embodiments, the pharmaceutically acceptable lyophilized excipient is selected from one or a mixture of two or more of the following: glucose, fructose, maltose, sorbitol, mannitol, xylitol, glycerol, sucrose, trehalose, lactose, alanine, cysteine, cystine, histidine, glutamic acid, phenylalanine, glycine, aspartic acid, lysine, leucine, arginine, serine, tryptophan, sodium chloride, zinc sulfate, EDTA, dextran, disodium hydrogen phosphate, sodium dihydrogen phosphate, sodium bicarbonate, sodium malate, sodium tartrate, sodium lactate, human serum albumin, hyaluronic acid, chitosan, alginate, hydroxyethyl starch, povidone, polyethylene glycol, polyvinyl alcohol, polylysine, polyaspartic acid, poloxamer, phospholipids, cholesterol, gelatin, collagen, protamine sulfate, etc. In some embodiments, the pharmaceutically acceptable lyophilization excipient is selected from one or more of mannitol, sucrose, trehalose, glucose, and sodium chloride, such as trehalose and mannitol.

[0017] On the other hand, the present invention provides a method for preparing the pharmaceutical composition of the present invention, the method comprising the following steps:

[0018] Step 1: Dissolve the active ingredient utidelon in an organic solvent to form an oil phase solution, and dissolve or dilute human serum albumin in an aqueous solution to form an aqueous phase solution, wherein the aqueous solution is a buffer salt solution;

[0019] Step 2: Add the oil phase solution of utidelon to the aqueous phase solution of human serum albumin to form an emulsion;

[0020] Step 3: Homogenize the emulsion from Step 2 into a nano-suspension using a high-pressure homogenizer;

[0021] Step 4: While homogenizing, rapidly remove the organic solvent from the nanosuspension obtained in Step 3 to obtain a suspension or powder containing albumin-bound utidylcholine nanoparticles; and

[0022] Optionally, step 5: freeze-dry the suspension obtained in step 4 to obtain the lyophilized powder form of the pharmaceutical composition of the present invention.

[0023] In some embodiments, in step 1 of the method, the organic solvent used to dissolve utidylcholine is selected from one or more of the following: chloroform, dichloroform, ethyl acetate, propyl acetate, butyl acetate, ethyl formate, methanol, ethanol, acetone, n-propanol, isopropanol, tert-butanol, methyl butyl ketone, and butanone. In some embodiments, in step 1 of the method, the organic solvent used to dissolve utidylcholine is a mixture of one or more of chloroform and dichloroform with one or more of methanol, ethanol, tert-butanol, and acetone. In some specific embodiments, the volume ratio of chloroform and / or dichloroform to methanol, ethanol, tert-butanol, and / or acetone is in the range of 20:1 to 1:1, for example, in the range of 15:1 to 5:1, for example, volume ratios of 9:1, 8:2, 7:3, 11:1, etc. In some embodiments, the organic solvent used to dissolve utedelon is a mixture of chloroform or dichloroform with methanol, ethanol, tert-butanol, or acetone. In some specific embodiments, the volume ratio of chloroform or dichloroform to methanol, ethanol, tert-butanol, or acetone is in the range of 20:1 to 1:1, for example, in the range of 15:1 to 5:1, such as volume ratios of 9:1, 8:2, 7:3, 11:1, etc.

[0024] In some embodiments, the organic solvent used to dissolve urothalon is a mixture of chloroform and ethanol in a ratio of 9:1 (v / v). In some embodiments, the organic solvent used to dissolve urothalon is a mixture of dichloroform and ethanol in a ratio of 9:1 (v / v). In some embodiments, the organic solvent used to dissolve urothalon is a mixture of chloroform and tert-butanol in a ratio of 9:1 (v / v). In some embodiments, the organic solvent used to dissolve urothalon is a mixture of chloroform and acetone in a ratio of 9:1 (v / v). In some embodiments, the organic solvent used to dissolve urothalon is a mixture of chloroform and methanol in a ratio of 9:1 (v / v).

[0025] In some embodiments, in step 1 of the method, the concentration of utidylcholine in the oil phase solution is in the range of 0.1 mg / ml to 5000 mg / ml, for example, in the range of 5 mg / mL to 2000 mg / ml, 20 mg / mL to 500 mg / ml, or 50 mg / mL to 300 mg / ml.

[0026] In some embodiments, in step 1 of the method, the aqueous solution used to dissolve or dilute human serum albumin is a buffer salt solution. The inventors found in experiments that the pH of the buffer salt has no significant effect on the stability and quality of the final product; however, the stability and quality of the product are related to the type of buffer solution. In some embodiments, the aqueous solution used to dissolve or dilute human serum albumin is a buffer salt solution with a pH of 4.0 to 8.0. In some embodiments, the buffer salt solution can be an inorganic acid salt, including but not limited to phosphates. In some embodiments, the buffer salt solution can be an organic acid salt, including but not limited to one or more of citrate, tartrate, acetate, fumarate, ethylenediaminetetraacetic acid (EDTA), and malate. In one embodiment, the buffer salt solution is selected from one or more of phosphate, citrate, tartrate, acetate, fumarate, EDTA, and malate solutions. In some specific embodiments, the buffer salt solution is selected from phosphate solutions. In some specific embodiments, the buffer salt solution is selected from one or more of citrate, malate, tartrate, fumarate, and EDTA solutions. In some embodiments, the buffer salt solution is selected from citrate, tartrate, and ethylenediaminetetraacetic acid solutions. In some embodiments, the buffer salt solution is selected from citrate solutions.

[0027] In some embodiments, in step 1 of the method, the buffer salt solution is a phosphate buffer with pH 4.0–8.0, such as a phosphate buffer with pH 4.5. In some embodiments, in step 1 of the method, the buffer salt solution is a citrate buffer with pH 4.0–8.0, such as a citrate buffer with pH 5.4, pH 6.8, or pH 7.2. In some embodiments, in step 1 of the method, the buffer salt solution is a malate buffer with pH 4.0–8.0, such as a malate buffer with pH 7.2. In some embodiments, in step 1 of the method, the buffer salt solution is a tartrate buffer with pH 4.0–8.0, such as a tartrate buffer with pH 7.2. In some embodiments, in step 1 of the method, the buffer salt solution is a fumarate buffer with pH 4.0–8.0, such as a fumarate buffer with pH 7.2. In some embodiments, in step 1 of the method, the buffer salt solution is an ethylenediaminetetraacetic acid buffer solution with a pH of 4.0 to 8.0, such as a sodium ethylenediaminetetraacetic acid buffer solution with a pH of 7.2.

[0028] In some embodiments, in step 1 of the method, the concentration of human serum albumin in the aqueous solution is in the range of 1 mg / ml to 1000 mg / ml, for example, in the range of 10 mg / ml to 200 mg / ml, 20 mg / ml to 150 mg / ml, or 30 mg / ml to 100 mg / ml.

[0029] In some embodiments, the weight ratio of the active ingredient utidylcholine to the carrier material human serum albumin in the method is in the range of 0.01% to 99.9%, for example, in the range of 80% to 5%, 75% to 15%, or 70% to 20%, for example, 5%, 10%, 20%, 25%, 33%, 40%, 44%, 50%, 60%, 67%, 80%, or any two of the above values.

[0030] In some embodiments, in step 2 of the method, a stirrer or shearing machine is used to mix the utidylcholine oil phase solution and the human serum albumin aqueous phase solution to form an emulsion.

[0031] In some embodiments, if necessary, the same buffer salt solution used in step 1 can be added to the emulsion in step 2 after step 2 of the method to increase the volume for homogenization.

[0032] In some embodiments, in step 3 of the method, the emulsion from step 2 is homogenized into a nanosuspension using a high-pressure homogenizer at a pressure of 5000 psi to 30000 psi, for example, 10000 psi to 25000 psi.

[0033] The inventors also studied the effect of the number of homogenization cycles on the particle size and stability of the final product, and found that different homogenization cycles had no significant effect on the particle size and stability of the final product.

[0034] In some embodiments, if necessary, an aqueous solution may be added to the nanosuspension of step 3 after step 3 of the method to dilute the nanosuspension to the desired concentration. In some embodiments, the aqueous solution is selected from one or more of water for injection, buffered saline solution, sucrose solution, mannitol solution, glucose solution, trehalose solution, etc. In some embodiments, the aqueous solution is water for injection. In some embodiments, the aqueous solution is a buffered saline solution, such as the buffered saline solution described in step 1 of the method.

[0035] In some embodiments, in step 4 of the method, the method for rapidly removing organic solvents includes removing organic solvents from the nanosuspension by means of rotary evaporation, thin-film evaporation with reduced pressure, and freeze spray drying.

[0036] In some embodiments, prior to the rapid removal of organic solvent in step 4 of the method, the method further includes feeding the nanosuspension obtained in step 3 into a vacuum chamber via a power source.

[0037] In some embodiments, the power source refers to a device capable of delivering the nano-suspension from step 3 into the vacuum chamber, such as a peristaltic pump, a syringe pump, or compressed air. In some embodiments, a peristaltic pump is used to pump the nano-suspension from step 3 into the vacuum chamber at a speed of 20 to 100 rpm while homogenizing.

[0038] In some embodiments, the vacuum chamber refers to a container or device capable of maintaining a certain degree of vacuum inside. Examples include, but are not limited to, rotary evaporators, thin-film evaporators, and freeze spray dryers. In some embodiments, the vacuum chamber is a rotary evaporator. In some embodiments, the vacuum chamber is a thin-film evaporator.

[0039] In some embodiments, depending on the type of vacuum chamber, suitable methods such as rotary evaporation, thin-film evaporation, or freeze-spray drying can be used to rapidly remove organic solvents. In some embodiments, organic solvents are rapidly removed using a reduced-pressure method in a thin-film evaporator, wherein the vacuum level of the thin-film evaporator is 1 mBar to 500 mBar, preferably 10 mBar to 200 mBar, and the scraper speed is 50 rpm to 300 rpm, preferably 100 rpm to 200 rpm.

[0040] Those skilled in the art will understand that when the organic solvent is removed from the nanoemulsion by freeze-spray drying in step 4 of the method, the subsequent freeze-drying step 5 is unnecessary.

[0041] In some embodiments, in step 5 of the method, the suspension obtained in step 4 may be dispensed into suitable containers for freeze-drying, with or without the addition of a lyophilizing excipient, such as the lyophilizing excipient described above. In some specific embodiments, in step 5 of the method, the suspension obtained in step 4 is diluted to a specified concentration with water for injection or a buffer salt solution, and then freeze-dried to obtain the lyophilized powder form of the pharmaceutical composition of the present invention.

[0042] In another aspect, the present invention provides a pharmaceutical composition prepared using the method described in the present invention.

[0043] In another aspect, the present invention provides a pharmaceutical composition for treating tumors such as advanced breast cancer, lung cancer, gastric cancer, liver cancer, and other solid tumors.

[0044] In another aspect, the present invention provides a method for treating a subject's tumor, such as advanced breast cancer, lung cancer, gastric cancer, liver cancer, or other solid tumors, the method comprising administering to the subject a therapeutically effective amount of the pharmaceutical composition described in the present invention.

[0045] In another aspect, the present invention provides the use of the pharmaceutical composition described herein in the preparation of a medicament for treating tumors such as advanced breast cancer, lung cancer, gastric cancer, liver cancer, and other solid tumors. In some embodiments, the medicament is an injection. Those skilled in the art can prepare the pharmaceutical composition described herein into an injection using conventional preparation methods in the pharmaceutical field. In other embodiments, the medicament is a sterile lyophilized powder for injection. Those skilled in the art can prepare the pharmaceutical composition described herein into a sterile lyophilized powder for injection using conventional preparation methods in the pharmaceutical field.

[0046] In another aspect, the present invention provides an injectable preparation comprising albumin-bound utidelon nanoparticles or pharmaceutical compositions as described herein.

[0047] In another aspect, the present invention provides an albumin-bound utidelon nanoparticle formulation for injection.

[0048] In another aspect, the present invention provides a sterile lyophilized powder injection comprising albumin-bound utidelon nanoparticles or pharmaceutical compositions as described in the present invention.

[0049] In another aspect, the present invention provides a sterile lyophilized powder injection of albumin-bound utederolene nanoparticles. In some embodiments, before administration, the albumin-bound utederolene nanoparticle sterile lyophilized powder injection is redissolved in a suitable liquid such as 5% glucose solution, 0.9% sodium chloride solution, or sterile water for injection, and then administered by a suitable method such as intravenous injection or intravenous drip.

[0050] The technical solution of the present invention has the following beneficial effects:

[0051] The albumin-bound utedelone nanoparticles or pharmaceutical compositions containing them of the present invention exhibit good physical and chemical stability under storage conditions of 2°C to 8°C and room temperature, thereby improving drug compliance and safety.

[0052] The preparation method of the present invention can make the drug loading of utidelon as high as 20% or even 60%, thereby making the total amount of albumin dimers, oligomers and polymers in the albumin-bound utidelon nanoparticles at a unit dose low, which is beneficial to improving the quality of the final product.

[0053] The preparation method of the present invention improves the stability of albumin-bound utedelone nanoparticle suspension at room temperature by dissolving human serum albumin in a specific buffer salt solution;

[0054] The preparation method of the present invention improves the stability of albumin-bound utedellon nanoparticles by controlling the desolventization rate and efficiency.

[0055] The albumin-bound utedelone nanoparticle suspension obtained by the preparation method of the present invention has good particle size uniformity and good filterability, making sterilization filtration in industrial production possible, and at the same time has a very high filtration yield, making it suitable for industrial production.

[0056] The albumin-bound utederolene nanoparticle lyophilized powder of the present invention can be redissolved in a suitable liquid such as 5% glucose solution or 0.9% sodium chloride solution to quickly form an albumin-bound utederolene nanoparticle suspension with relatively uniform particle size. This suspension has good stability at room temperature and can meet the needs of clinical applications. Attached Figure Description

[0057] Figure 1 Particle size distribution diagram of albumin-bound utidelon nanoparticles in Example 10. Detailed Implementation

[0058] The present invention will be further described below with reference to specific embodiments. However, the present invention is not limited to these embodiments. For those skilled in the art, equivalent substitutions, combinations, improvements or modifications can be made to the present invention without departing from the principle of the present invention, and these will all be included within the protection scope of the present invention.

[0059] In this specific embodiment, the 20% human serum albumin solution was obtained from Chengdu Rongsheng Pharmaceutical Co., Ltd., and Utidelon was obtained from Chengdu Huahao Zhongtian Pharmaceutical Co., Ltd. All other reagents were commercially available.

[0060] Comparative Example 1

[0061] Take 7.5 ml of 20% human serum albumin solution, dilute with water for injection to 30 ml, add 5 ml of 200 mg / ml utedrol solution (solvent: chloroform:ethanol = 9:1 (v / v)), and shear into a colostrum using a high-speed shear machine (IKA model T25) at 5000 rpm. Then, homogenize using a homogenizer (Shanghai Nuozhe, model Nano) at 25000 psi for 6 cycles. Take 4 aliquots, approximately 1 ml each, and sequentially transfer them to a rotary evaporator (Swiss Buzaki, model R-300). Remove the organic solvent from the solution under evaporation conditions of 40℃ water bath heating and 40 mbar reduced pressure, obtaining samples Y1, Y2, Y3, and Y4. The time interval for solvent removal from samples Y1, Y2, Y3, and Y4 was approximately 10 minutes. The nanoparticle size of samples Y1, Y2, Y3, and Y4 gradually increased over time, as shown in the table below.

[0062]

[0063] After the rotary evaporation of the four samples was completed, the remaining suspension was placed in a rotary evaporator (Swiss Buzaki, model R-300) and heated in a water bath at 40°C. The organic solvent in the solution was removed under reduced pressure evaporation conditions of 40 mbar. During the rotary evaporation process, precipitate was visible to the naked eye.

[0064] Comparative Example 2

[0065] Take 27 ml of 20% human serum albumin solution, dilute it to 60 ml with water for injection, add 9 ml of 200 mg / ml utederosol solution (solvent: chloroform: ethanol = 9:1 (v / v)), and shear it into colostrum using a high-speed shear machine (IKA model T25) at 8000 rpm. Then, homogenize it for 6 cycles using a homogenizer (Shanghai Nuozhe, model Nano) at a pressure of 20000 psi. While homogenizing, pump the solution into a thin-film evaporator (Shanghai Deda Tianyi, model SH-DEA) at 55 rpm using a peristaltic pump to remove organic solvent. The vacuum degree of the thin-film evaporator is 1500 Pa and the scraper speed is 150 rpm. After evaporation, rinse the tubing and the thin-film evaporator with about 150 ml of water for injection to obtain a suspension containing albumin-bound utederosol nanoparticles. The average diameter of albumin-bound utedelone nanoparticles in the suspension was measured to be 128.16 nm using a Nicomp Z3000 nanoparticle size and Zeta potential analyzer from PSS Corporation, USA.

[0066] Example 1

[0067] Take 9 ml of 20% human serum albumin solution, add pH 4.5 phosphate buffer to 60 ml, mix well, add 2.5 ml of 200 mg / ml utidylcholine solution (solvent: chloroform:ethanol = 9:1 (v / v)), and shear into a primary emulsion using a high-speed shearing machine (IKA, model T25) at 5000 rpm. Then homogenize using a homogenizer (Shanghai Nuozhe, model Nano) at 25000 psi for 4, 6, 8, and 10 cycles. Take a small amount (approximately 0.5 ml) of each nanoemulsion and transfer it to a rotary evaporator (Swiss Buzaki, model R-300). Heat in a 40℃ water bath and evaporate under reduced pressure at 40 mbar to remove the organic solvent, obtaining a suspension containing albumin-bound utidylcholine nanoparticles. The average diameter of the albumin-bound utidylcholine nanoparticles in the suspension was measured using a Nicomp Z3000 nanoparticle size and Zeta potential analyzer from PSS (USA). The results are shown in the table below.

[0068]

[0069] The results of this example demonstrate that, compared to Comparative Example 1, using phosphate buffer instead of water for injection to dissolve human serum albumin can form stable albumin-bound utidelon nanoparticles. The results of this example also show that different homogenization times have no significant effect on the particle size and product quality of the final nanoparticle product.

[0070] Example 2

[0071] Take 6 ml of 20% human serum albumin solution, add phosphate buffer solutions of different pH values ​​to a final volume of 30 ml, mix well, add 3 ml of 100 mg / ml utidylcholine solution (solvent: chloroform:ethanol = 9:1 (v / v)), and shear the mixture at 5000 rpm using a high-speed shearing machine (IKA, model T25) to form a primary emulsion. Then add 30 ml of the corresponding phosphate buffer solutions of different pH values ​​and homogenize four times using a homogenizer (Shanghai Nuozhe, model Nano) at 25000 psi. Transfer a small amount of nanoemulsion (approximately 0.5 ml) to a rotary evaporator (Swiss Buzaki, model R-300), heat in a 40°C water bath, and evaporate under reduced pressure at 40 mbar to remove the organic solvent, obtaining a suspension containing albumin-bound utidylcholine nanoparticles. The average diameter of the albumin-bound utidylcholine nanoparticles in the suspension was measured using a Nicomp Z3000 nanoparticle size and Zeta potential analyzer from PSS (USA). The results are shown in the table below.

[0072]

[0073] The results of this example demonstrate that, compared to Comparative Example 1, using phosphate buffer instead of water for injection to dissolve human serum albumin can form stable albumin-bound utidylcholine nanoparticles. The results also indicate that phosphate buffer solutions of different pH values ​​have no significant impact on the particle size and quality of the final nanoparticle product.

[0074] Example 3

[0075] Take 7.5 ml of 20% human serum albumin solution, dilute it to 30 ml with pH 4.5 phosphate buffer, mix well, add 3 ml of 200 mg / ml utederolide solution (solvent: chloroform: ethanol = 9:1 (v / v)), and shear it into colostrum using a high-speed shearing machine (IKA, model T25) at 5000 rpm. Add 30 ml of pH 4.5 phosphate buffer, homogenize 4 times with a homogenizer (Shanghai Nuozhe, model Nano) at 25000 psi, then add about 250 ml of pH 4.5 phosphate buffer. While homogenizing, pump the solution into a thin-film evaporator (Shanghai Deda Tianyi, model SH-DEA) at 65 rpm using a peristaltic pump to remove organic solvents. The vacuum degree of the thin-film evaporator is 1500 Pa and the scraper speed is 150 rpm. After evaporation, a suspension containing albumin-bound utederolide nanoparticles is obtained. The average diameter of albumin-bound utedelone nanoparticles in the suspension was measured using a Nicomp Z3000 nanoparticle size and Zeta potential analyzer from PSS Corporation, and was found to be 125 nm.

[0076] Example 4

[0077] Take 9 ml of 20% human serum albumin solution, dilute it to 60 ml with pH 4.5 phosphate buffer, add 6 ml of 200 mg / ml utederolide solution (solvent: chloroform: ethanol = 9:1 (v / v)), and shear it into colostrum using a high-speed shear machine (IKA model T25) at 5000 rpm. Then, homogenize it for 4 cycles using a homogenizer (Shanghai Nuozhe, model Nano) at a pressure of 25000 psi. Add about 250 ml of water for injection, and while homogenizing, pump it into a thin-film evaporator (Shanghai Deda Tianyi, model SH-DEA) at 55 rpm using a peristaltic pump to remove the organic solvent. The vacuum degree of the thin-film evaporator is 1500 Pa and the scraper speed is 150 rpm. After evaporation, a suspension containing albumin-bound utederolide nanoparticles is obtained. The average diameter of albumin-bound utedelone nanoparticles in the suspension was measured using a Nicomp Z3000 nanoparticle size and Zeta potential analyzer from PSS Corporation, USA, and was found to be 134 nm.

[0078] Example 5

[0079] Take 9 ml of 20% human serum albumin solution, dilute it to 60 ml with pH 4.5 phosphate buffer, add 4 ml of 200 mg / ml utederosol solution (solvent: dichloromethane: ethanol = 9:1 (v / v)), and shear it into colostrum using a high-speed shear machine (IKA model T25) at 5000 rpm. Then, homogenize it for 4 cycles using a homogenizer (Shanghai Nuozhe, model Nano) at a pressure of 25000 psi. Add about 200 ml of water for injection, and while homogenizing, pump it into a thin-film evaporator (Shanghai Deda Tianyi, model SH-DEA) at 55 rpm using a peristaltic pump to remove the organic solvent. The vacuum degree of the thin-film evaporator is 1500 Pa and the scraper speed is 150 rpm. After evaporation, a suspension containing albumin-bound utederosol nanoparticles is obtained. The average diameter of albumin-bound utedelone nanoparticles in the suspension was measured using a Nicomp Z3000 nanoparticle size and Zeta potential analyzer from PSS Corporation, USA, and was found to be 135 nm.

[0080] Example 6

[0081] Take 9 ml of 20% human serum albumin solution, dilute it to 60 ml with pH 4.5 phosphate buffer, add 4 ml of 200 mg / ml utederolide solution (solvent: chloroform: tert-butanol = 9:1 (v / v)), and shear it into colostrum using a high-speed shear machine (IKA model T25) at 5000 rpm. Then, homogenize it for 4 cycles using a homogenizer (Shanghai Nuozhe, model Nano) at a pressure of 25000 psi. Add about 200 ml of water for injection, and while homogenizing, pump it into a thin-film evaporator (Shanghai Deda Tianyi, model SH-DEA) at 55 rpm using a peristaltic pump to remove organic solvents. The vacuum degree of the thin-film evaporator is 1500 Pa and the scraper speed is 150 rpm. After evaporation, a suspension containing albumin-bound utederolide nanoparticles is obtained. The average diameter of albumin-bound utedelone nanoparticles in the suspension was measured using a Nicomp Z3000 nanoparticle size and Zeta potential analyzer from PSS Corporation, USA, and was found to be 155 nm.

[0082] Example 7

[0083] Take 9 ml of 20% human serum albumin solution, dilute it to 60 ml with pH 4.5 phosphate buffer, add 4 ml of 200 mg / ml utederolide solution (solvent: chloroform: acetone = 9:1 (v / v)), and shear it into colostrum using a high-speed shear machine (IKA model T25) at 5000 rpm. Then, homogenize it for 4 cycles using a homogenizer (Shanghai Nuozhe, model Nano) at a pressure of 25000 psi. Add about 200 ml of water for injection, and while homogenizing, pump it into a thin-film evaporator (Shanghai Deda Tianyi, model SH-DEA) at 55 rpm using a peristaltic pump to remove organic solvent. The vacuum degree of the thin-film evaporator is 1500 Pa and the scraper speed is 150 rpm. After evaporation, a suspension containing albumin-bound utederolide nanoparticles is obtained. The average diameter of albumin-bound utedelone nanoparticles in the suspension was measured using a Nicomp Z3000 nanoparticle size and Zeta potential analyzer from PSS Corporation, USA, and was found to be 135 nm.

[0084] Example 8

[0085] Take 9 ml of 20% human serum albumin solution, dilute it to 60 ml with pH 4.5 phosphate buffer, add 4 ml of 200 mg / ml utederolide solution (solvent: chloroform:methanol = 9:1 (v / v)), and shear it into colostrum using a high-speed shear machine (IKA model T25) at 5000 rpm. Then, homogenize it for 4 cycles using a homogenizer (Shanghai Nuozhe, model Nano) at a pressure of 25000 psi. Add about 200 ml of water for injection, and while homogenizing, pump it into a thin-film evaporator (Shanghai Deda Tianyi, model SH-DEA) at 55 rpm using a peristaltic pump to remove organic solvent. The vacuum degree of the thin-film evaporator is 1500 Pa and the scraper speed is 150 rpm. After evaporation, a suspension containing albumin-bound utederolide nanoparticles is obtained. The average diameter of albumin-bound utedelone nanoparticles in the suspension was measured using a Nicomp Z3000 nanoparticle size and Zeta potential analyzer from PSS Corporation, USA, and was found to be 127 nm.

[0086] Example 9

[0087] Take 9 ml of 20% human serum albumin solution, dilute it to 60 ml with pH 4.5 phosphate buffer, add 4.5 ml of 200 mg / ml utederolide solution (solvent: chloroform: ethanol = 9:1 (v / v)), and shear it into colostrum using a high-speed shear machine (IKA model T25) at 5000 rpm. Then, homogenize it for 4 cycles using a homogenizer (Shanghai Nuozhe, model Nano) at a pressure of 25000 psi. Add about 200 ml of water for injection, and while homogenizing, pump it into a thin-film evaporator (Shanghai Deda Tianyi, model SH-DEA) at 55 rpm using a peristaltic pump to remove organic solvent. The vacuum degree of the thin-film evaporator is 1500 Pa and the scraper speed is 150 rpm. After evaporation, a suspension containing albumin-bound utederolide nanoparticles is obtained. The average diameter of albumin-bound utedelone nanoparticles in the suspension was measured using a Nicomp Z3000 nanoparticle size and Zeta potential analyzer from PSS Corporation, USA, and was found to be 130 nm.

[0088] Example 10

[0089] Take 18 ml of 20% human serum albumin solution, dilute it to 60 ml with pH 6.8 citrate buffer, add 3.6 ml of 200 mg / ml utederolide solution (solvent: chloroform: ethanol = 9:1 (v / v)), and shear it into colostrum using a high-speed shear machine (IKA model T25) at 5000 rpm. Then, homogenize it for 5 cycles using a homogenizer (Shanghai Nuozhe, model Nano) at a pressure of 25000 psi. Add about 200 ml of pH 6.8 citrate buffer, and while homogenizing, pump it into a thin-film evaporator (Shanghai Deda Tianyi, model SH-DEA) at 55 rpm using a peristaltic pump to remove organic solvent. The vacuum degree of the thin-film evaporator is 1500 Pa and the scraper speed is 150 rpm. After evaporation, a suspension containing albumin-bound utederolide nanoparticles is obtained. The average diameter of albumin-bound utedelone nanoparticles in the suspension was measured using a Nicomp Z3000 nanoparticle size and Zeta potential analyzer from PSS Corporation, USA, and was found to be 132 nm.

[0090] Example 11

[0091] Take 9 ml of 20% human serum albumin solution, dilute it to 60 ml with pH 5.4 citrate buffer, add 2.25 ml of 200 mg / ml utederolide solution (solvent: chloroform: ethanol = 9:1 (v / v)), and shear it into colostrum using a high-speed shear machine (IKA model T25) at 5000 rpm. Then, homogenize it for 6 cycles using a homogenizer (Shanghai Nuozhe, model Nano) at a pressure of 25000 psi. Add about 200 ml of pH 5.4 citrate buffer, and while homogenizing, pump it into a thin-film evaporator (Shanghai Deda Tianyi, model SH-DEA) at 55 rpm using a peristaltic pump to remove organic solvent. The vacuum degree of the thin-film evaporator is 1500 Pa and the scraper speed is 150 rpm. After evaporation, a suspension containing albumin-bound utederolide nanoparticles is obtained. The average diameter of albumin-bound utedelone nanoparticles in the suspension was measured using a Nicomp Z3000 nanoparticle size and Zeta potential analyzer from PSS Corporation, USA, and was found to be 128 nm.

[0092] Example 12

[0093] Take 27 ml of 20% human serum albumin solution, dilute it to 60 ml with the buffer solution described in the table below, add 9 ml of 200 mg / ml utederolide solution (solvent is chloroform:ethanol = 9:1 (v / v)), and shear it into colostrum using a high-speed shear machine (IKA model T25) at 8000 rpm. Then, homogenize it for 6 cycles using a homogenizer (Shanghai Nuozhe, model Nano) at a pressure of 20000 psi. While homogenizing, pump it into a thin-film evaporator (Shanghai Deda Tianyi, model SH-DEA) at a speed of 55 rpm to remove organic solvent. The vacuum degree of the thin-film evaporator is 1500 Pa and the scraper speed is 150 rpm. After evaporation, rinse the tubing and the thin-film evaporator with about 150 ml of the corresponding buffer salt solution to obtain a suspension containing albumin-bound utederolide nanoparticles. The average diameter of albumin-bound utedelone nanoparticles in the suspension was measured using a Nicomp Z3000 nanoparticle size and Zeta potential analyzer from PSS Corporation, USA. The results are shown in the table below.

[0094]

[0095] The results of this example show that, in addition to inorganic acid salt buffers such as phosphate, stable albumin-bound utidylcholine nanoparticles can also be obtained using various organic acid salt buffers.

[0096] Example 13

[0097] Approximately 15 ml of albumin-bound utedelone nanoparticle suspensions from Comparative Examples 2, 8, and 10 were dispensed into vials and placed at 2°C to 8°C to investigate the particle size stability of the nanoparticle suspensions. The specific results are shown in the table below.

[0098]

[0099] The results showed that, under storage conditions of 2℃ to 8℃, the nanoparticle size of albumin obtained by dissolving albumin with water for injection as the aqueous phase in Comparative Example 2 gradually increased over time, while the albumin nanoparticles obtained by using buffer salt solution as the aqueous phase in this invention had good particle size stability.

[0100] Example 14

[0101] Approximately 15 ml of albumin-bound utidylcholine nanoparticle suspensions from Comparative Examples 2, 8, and 10 were dispensed into vials and placed at 25°C to investigate the particle size stability of the nanoparticle suspensions. The specific results are shown in the table below.

[0102]

[0103]

[0104] The results showed that, in Comparative Example 2, the nanoparticles quickly aggregated and precipitated at room temperature. However, when a buffer salt solution was used as the aqueous phase, the stability of the nanoparticles was better than that of water for injection. Among them, the particle size stability of organic acid salts such as citrate buffer (Example 10) was better than that of inorganic acid salts such as phosphate buffer (Example 8).

[0105] Example 15

[0106] Take an appropriate amount of albumin-bound utedelone nanoparticle suspension from Examples 8 and 10, add an appropriate amount of lyophilization excipient, dilute with water for injection to approximately 5 mg of utedelone per ml, and quantitatively dispense into vials. Place in a freeze dryer, rapidly cool the solution to -40°C within 1 hour, maintain this temperature for 3–4 hours to obtain a frozen body, evacuate to a vacuum degree less than 300 Pa, then raise the temperature to -20°C within 1–2 hours and maintain for 20–48 hours, then raise the temperature to 5°C within 1–2 hours and maintain for 2–4 hours, then raise the temperature to 25°C within 1–2 hours and maintain for 2–4 hours, stopper, and remove from the freeze dryer. After freeze-drying, resuspend in 0.9% sodium chloride solution, and measure the particle size using a Nicomp Z3000 nanoparticle size and Zeta potential analyzer from PSS Corporation (USA). The test results are shown in the table below.

[0107]

[0108] Example 16

[0109] Take an appropriate amount of the freeze-dried powder from Comparative Example 2, Example 8, Example 10 and Example 12 above, dissolve it in 5% glucose solution, let it stand at 2℃~8℃ for 30 minutes, and then transfer it to a 25℃ drug stability test chamber to investigate the particle size stability of albumin-bound utedelone nanoparticles at room temperature after freeze-drying. The specific results are shown in the table below.

[0110]

[0111]

[0112] The results showed that the nanoparticles of Comparative Example 2 quickly aggregated and precipitated at room temperature, while the nanoparticles of Examples 10 and 12 showed a particle size change of less than 10 nm within 4 hours at room temperature, indicating good stability and suggesting good feasibility for clinical application.

[0113] Example 17

[0114] In this embodiment, the filterability of the albumin-bound utidelon nanoparticle suspension described in Example 10 was evaluated by passing it through a 0.45 μm filter in series with a 0.22 μm filter. The filterability of the suspension was evaluated by the change in utidelon content before and after filtration.

[0115] Take an appropriate amount of the albumin-bound utidylcholine nanoparticle suspension from Example 10, filter it using a 0.45 μm syringe filter connected in series with a 0.22 μm syringe filter, and place 1 ml of each of the nanoparticle suspension before and after filtration into a 10 ml volumetric flask. Add an appropriate amount of acetonitrile, and sonicate for about 10 minutes to denature and precipitate the albumin. Dilute to the mark with acetonitrile, mix well, centrifuge at high speed, and use the supernatant as the test solution to measure the utidylcholine content. The utidylcholine content determination method uses HPLC, with a detection wavelength of 250 nm, a C18 column, and a mobile phase of acetonitrile-water (60:40, v / v). The injection volume is 10 μl. The test results show that the albumin-bound utidylcholine nanoparticle suspension from Example 10 has good filterability, and the change rate of utidylcholine content before and after filtration is less than 5%. This indicates that the albumin-bound utidylcholine nanoparticles prepared by this invention have good particle size uniformity and good filterability, making sterilization filtration possible in industrial production. At the same time, they have a very high filtration yield and are suitable for industrial production.

[0116] Example 18

[0117] In this embodiment, the contents of albumin dimers, oligomers, and polymers in the albumin-bound utedelone nanoparticle suspension described in Example 10 were evaluated.

[0118] Take an appropriate amount of the lyophilized albumin-bound utidylcholine nanoparticle powder from Example 10, dilute it with water for injection to approximately 10 mg of human serum albumin per ml, filter it using a 0.45 μm syringe filter followed by a 0.22 μm syringe filter, and use the filtrate as the test solution to measure the content of albumin dimers, oligomers, and polymers in the test solution. The content of albumin dimers, oligomers, and polymers in albumin-bound utidylcholine nanoparticles was determined by HPLC. The chromatographic column was a hydrophilic silica high-performance size exclusion column, the mobile phase was a pH 7.0, 0.2 mol / L phosphate buffer containing 1% isopropanol, the detection wavelength was 280 nm, the flow rate was 0.6 ml / min, and the injection volume was 20 μl. Test results show that the total amount of albumin oligomers, dimers, and polymers in a single dose of albumin-bound utedelone nanoparticles (30 mg / vial, calculated as utedelone) in Example 10 does not exceed 15 mg, which is far less than the limit in human serum albumin injection (500 mg albumin polymers / vial). This example demonstrates that the total amount of albumin dimers, oligomers, and polymers in the albumin-bound utedelone nanoparticles prepared by this invention at a unit dose is low, which is beneficial to improving the quality of the final product, thereby improving drug compliance and safety.

Claims

1. A pharmaceutical composition comprising albumin-bound utedelone nanoparticles, said nanoparticles comprising the active ingredient utedelone, human serum albumin, and a pharmaceutically acceptable buffer salt.

2. The pharmaceutical composition according to claim 1, wherein the active ingredient utidelon is encapsulated or adsorbed onto human serum albumin.

3. The pharmaceutical composition according to claim 1 or 2, wherein the pharmaceutically acceptable buffer salt is selected from one or more of phosphates, citrates, acetates, tartrates, malates, fumarates, and ethylenediaminetetraacetic acid salts.

4. The pharmaceutical composition according to claim 1 or 2, wherein the nanoparticles further comprise one or more of sucrose, mannitol, glucose, and trehalose.

5. The pharmaceutical composition according to claim 1 or 2, wherein the average particle size of the nanoparticles is in the range of 30 to 200 nm.

6. The pharmaceutical composition according to claim 1 or 2, wherein the pharmaceutical composition is in the form of a lyophilized powder.

7. The pharmaceutical composition of claim 6, wherein the pharmaceutical composition further comprises a pharmaceutically acceptable lyophilized excipient.

8. The pharmaceutical composition according to claim 7, wherein the pharmaceutically acceptable lyophilized excipient is selected from one or more of glucose, fructose, maltose, sorbitol, mannitol, xylitol, glycerol, sucrose, trehalose, lactose, alanine, cysteine, cystine, histidine, glutamic acid, phenylalanine, glycine, aspartic acid, lysine, leucine, arginine, serine, tryptophan, sodium chloride, zinc sulfate, EDTA, dextran, disodium hydrogen phosphate, sodium dihydrogen phosphate, sodium bicarbonate, sodium malate, sodium tartrate, sodium lactate, human serum albumin, hyaluronic acid, chitosan, alginate, hydroxyethyl starch, povidone, polyethylene glycol, polyvinyl alcohol, polylysine, polyaspartic acid, poloxamer, phospholipids, cholesterol, gelatin, collagen, and protamine sulfate.

9. A method for preparing the pharmaceutical composition according to any one of claims 1 to 8, comprising: The active ingredient, utidylcholine, is dissolved in an organic solvent to form an oil phase solution, and human serum albumin is dissolved or diluted in an aqueous solution to form an aqueous phase solution, wherein the aqueous solution is a buffer salt solution. The oil phase solution of utidelon was added to the aqueous phase solution of human serum albumin to form an emulsion; The emulsion was homogenized under high pressure using a homogenizer to form a nano suspension. The organic solvent is rapidly removed from the nanosuspension while homogenizing it to obtain a suspension or powder containing albumin-bound utidylcholine nanoparticles; and Optionally, the suspension is freeze-dried.

10. The method according to claim 9, wherein the organic solvent used to dissolve utedelon is selected from one or a mixture of two or more of the following solvents: chloroform, dichloromethane, ethyl acetate, propyl acetate, butyl acetate, ethyl formate, methanol, ethanol, acetone, n-propanol, isopropanol, tert-butanol, methyl butyl ketone, and butanone.

11. The method of claim 9, wherein the organic solvent used to dissolve utedelon is a mixture of chloroform or dichloroform with methanol, ethanol, tert-butanol or acetone.

12. The method according to claim 11, wherein the volume ratio of chloroform or dichloromethane to methanol, ethanol, tert-butanol or acetone is in the range of 20:1 to 1:

1.

13. The method of claim 12, wherein the volume ratio of chloroform or dichloromethane to methanol, ethanol, tert-butanol or acetone is in the range of 15:1 to 5:

1.

14. The method according to claim 9, wherein the concentration of utidelon in the oil phase solution is in the range of 0.1 mg / ml to 5000 mg / ml.

15. The method according to claim 14, wherein the concentration of utidelon in the oil phase solution is in the range of 5 mg / ml to 2000 mg / ml.

16. The method according to claim 14, wherein the concentration of utidelon in the oil phase solution is in the range of 20 mg / ml to 500 mg / ml.

17. The method according to claim 14, wherein the concentration of utidelon in the oil phase solution is in the range of 50 mg / mL to 300 mg / mL.

18. The method of claim 9, wherein the buffer salt solution is selected from one or more of phosphate buffer, citrate buffer, acetate buffer, tartrate buffer, fumarate buffer, ethylenediaminetetraacetic acid (EDTA) and malate solution.

19. The method according to claim 9, wherein the concentration of human serum albumin in the aqueous solution is in the range of 1 mg / ml to 1000 mg / ml.

20. The method according to claim 19, wherein the concentration of human serum albumin in the aqueous solution is in the range of 10 mg / ml to 200 mg / ml.

21. The method according to claim 19, wherein the concentration of human serum albumin in the aqueous solution is in the range of 20 mg / ml to 150 mg / ml.

22. The method according to claim 19, wherein the concentration of human serum albumin in the aqueous solution is in the range of 30 mg / ml to 100 mg / ml.

23. The method according to claim 9, wherein the weight ratio of the active ingredient utidelon to human serum albumin is in the range of 0.01% to 99.9%.

24. The method according to claim 23, wherein the weight ratio of the active ingredient utidelon to human serum albumin is in the range of 80% to 5%.

25. The method according to claim 23, wherein the weight ratio of the active ingredient utidelon to human serum albumin is in the range of 75% to 15%.

26. The method according to claim 23, wherein the weight ratio of the active ingredient utidelon to human serum albumin is in the range of 70% to 20%.

27. The method according to any one of claims 9 to 26, further comprising adding an aqueous solution to the nanosuspension after homogenizing it.

28. The method according to claim 27, wherein the aqueous solution is selected from one or more of water for injection, buffered salt solution, sucrose solution, mannitol solution, glucose solution and trehalose solution.

29. The method of claim 28, wherein the buffer salt solution is selected from one or more of phosphate buffer, citrate buffer, acetate buffer, tartrate buffer, fumarate buffer, ethylenediaminetetraacetic acid (EDTA) and malate solution.

30. The method of claim 9, wherein rapid removal of organic solvents is achieved by rotary evaporation, thin-film evaporation, or spray drying.

31. Use of the pharmaceutical composition of any one of claims 1 to 8 in the preparation of a medicament for treating or preventing cancer.

32. An injectable preparation comprising the pharmaceutical composition according to any one of claims 1 to 8.

33. A sterile lyophilized powder injection comprising the pharmaceutical composition according to any one of claims 1 to 8.

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