Plinabulin micelle composition and preparation method thereof

Abstract

The invention relates to a plinabulin micelle composition and a preparation method thereof, specifically, the plinabulin micelle composition is basically composed of plinabulin, 15-hydroxystearic acid polyethylene glycol ester and propylene glycol, and at least a part of plinabulin is encapsulated in a micelle. The Plinabulin micelle can be mixed with a glucose injection to obtain a Plinabulin micelle preparation for injection, and the Plinabulin micelle preparation can be used for clinically treating tumor diseases. The plinabulin micelle composition and the plinabulin micelle preparation have excellent electrochemical stability, dilution stability and storage stability.

Description

A Plinabulin micelle composition and preparation method thereof Technical Field

[0001] The present invention relates to the field of pharmaceutical preparations, and in particular to a Plinabulin micelle composition and a preparation method thereof. Background Art

[0002] Plinabulin ((3Z,6Z)-3-[(5-tert-butyl-1H-imidazol-4-yl)methylene]-6-(benzylidene)-2,5-piperazinedione) is a synthetic analog of the diketopiperazine phenylahistin (phenyltrimethylammonium chloride) found in marine and terrestrial Aspergillus species. Its structure is shown below:

[0003] Plinabulin is structurally different from colchicine and its combretastatin-like analogs (e.g., combretastatin phosphate) and binds at or near the colchicine binding site on tubulin monomers. Previous studies have shown that Plinabulin at low concentrations induces vascular endothelial cell tubulin depolymerization and monolayer permeability compared to colchicine, and that Plinabulin induces apoptosis in Jurkat leukemia cells. Studies of Plinabulin as a single agent in patients with advanced malignancies (lung cancer, prostate cancer, and colon cancer) have shown good pharmacokinetics, pharmacodynamics, and safety characteristics. However, there is still a need to provide an injectable formulation of Plinabulin with appropriate physicochemical properties.

[0004] Summary of the Invention

[0005] The present invention aims to provide a novel Plinabulin micelle and a preparation method thereof. The Plinabulin micelle has good stability. The obtained micelle can not only improve the water solubility of Plinabulin, but also delay drug release and prolong the biological half-life, and has good clinical application value.

[0006] The first aspect of the present invention provides a method for preparing a Plinabulin micellar composition, wherein the method comprises the steps of:

[0007] s1) mixing polyethylene glycol 15-hydroxystearate and propylene glycol at 35° C.-65° C. to prepare a clear mixture;

[0008] s2) mixing the clarified mixed solution in step 1) with Plinabulin at 35° C.-65° C. and stirring to prepare a Plinabulin micelle composition.

[0009] In another preferred embodiment, the weight ratio of 15-hydroxystearate polyethylene glycol to propylene glycol is 1:5-5:1, preferably, 1:3-3:1, and more preferably 2:3.

[0010] In another preferred embodiment, the weight ratio of polyethylene glycol 15-hydroxystearate to propylene glycol is 1:9, 2:8, 3:7, 4:6, 5:5, 6:4, 7:3, 8:2, or 9:1.

[0011] In another preferred embodiment, the micelle composition is essentially composed of Plinabulin, 15-hydroxystearate polyethylene glycol and propylene glycol.

[0012] In another preferred embodiment, in step s1), the temperature is 40°C-60°C.

[0013] In another preferred embodiment, in step s2), the temperature is 40°C-60°C.

[0014] In another preferred embodiment, step s1) is to melt 15-hydroxystearate polyethylene glycol at 50°C±5°C, then add propylene glycol, maintain the temperature at 40°C±5°C, and stir (for example, 5 min-2 h, preferably 20-40 min, more preferably 30 min) to prepare a clear mixed solution.

[0015] In another preferred embodiment, step s1) is to melt 15-hydroxystearate polyethylene glycol at 35°C-65°C, then add propylene glycol, maintain the temperature at 35°C-65°C, and stir (for example, 5 min-2 h, preferably 10-40 min, more preferably 20-30 min) to prepare a clear mixed solution.

[0016] In another preferred embodiment, step s1) is to melt 15-hydroxystearate polyethylene glycol at 35°C-65°C, add it to propylene glycol, maintain the temperature at 35°C-65°C, and stir (for example, 5 min-2 h, preferably 10-40 min, more preferably 20-30 min) to prepare a clear mixed solution.

[0017] In another preferred embodiment, the clarified mixed solution in step s2) is added in batches, preferably in 2-5 batches.

[0018] In another preferred embodiment, the concentration of Plinabulin is 0.02 mg / ml-4 mg / ml, based on the total mass of the Plinabulin micelle composition.

[0019] In another preferred embodiment, Plinabulin is Plinabulin anhydrate or Plinabulin monohydrate.

[0020] In another preferred embodiment, Plinabulin is Plinabulin anhydrate (crystalline form III) or Plinabulin monohydrate (crystalline form I).

[0021] In another preferred embodiment, in step (s2), the Plinabulin used is Plinabulin monohydrate.

[0022] In another preferred embodiment, Plinabulin is Plinabulin monohydrate (crystalline form I).

[0023] In another preferred embodiment, the Plinabulin anhydrate is the anhydrous form of Plinabulin in CN113735834B.

[0024] In another preferred embodiment, the Plinabulin monohydrate is the Plinabulin monohydrate in CN113735834B.

[0025] In another preferred embodiment, the particle size D50 of the micelles in the obtained Plinabulin micelle composition is 5-100 nm.

[0026] In a second aspect of the present invention, a Plinabulin micelle composition is provided, wherein the micelle composition comprises Plinabulin and a clear mixture of 15-hydroxystearate polyethylene glycol and propylene glycol, wherein the Plinabulin micelle composition is a yellow clear and transparent solution, and the micelle particle size ranges from 10 to 100 nm.

[0027] In another preferred embodiment, the micelle particle size of the Plinabulin micelle composition is in the range of 80-110 nm, preferably 90-110 nm, such as 100-110 nm.

[0028] In another preferred embodiment, the micelle D50 of the Plinabulin micelle composition is 80-110 nm, preferably 90-110 nm, such as 100-105 nm.

[0029] In another preferred embodiment, the micelle D90 of the Plinabulin micelle composition is 100-170 nm, preferably 110-160 nm, such as 120-150 nm.

[0030] In another preferred embodiment, in the Plinabulin micelle composition, the concentration of Plinabulin is 1 mg / ml-10 mg / ml. Based on the total mass of the Plinabulin micelle composition, the concentration of Plinabulin is preferably 2 mg / ml-5 mg / ml, and more preferably 3 mg / ml-4 mg / ml.

[0031] In another preferred embodiment, in the Plinabulin micelle composition, the Plinabulin is Plinabulin monohydrate.

[0032] In another preferred example, in the Plinabulin micelle composition, the weight ratio of 15-hydroxystearate polyethylene glycol to propylene glycol is 1:5-5:1, preferably, 1:3-3:1, and more preferably 2:3.

[0033] In another preferred embodiment, the Plinabulin micelle composition is prepared by the method described in the first aspect.

[0034] In another preferred embodiment, the Plinabulin micelle composition is sterile.

[0035] The third aspect of the present invention provides a liquid Plinabulin composition for injection, comprising:

[0036] Plinabulin, propylene glycol, and polyethylene glycol 15-hydroxystearate D5W solution (5% glucose injection);

[0037] The volume ratio of propylene glycol to D5W in the composition is about 6:50 to about 6:500.

[0038] In another preferred embodiment, in the liquid Plinabulin composition for injection, the Plinabulin is Plinabulin monohydrate.

[0039] In another preferred embodiment, Plinabulin is encapsulated in micelles, preferably, at least 90% of Plinabulin is encapsulated in micelles.

[0040] In another preferred embodiment, at least 95% of Plinabulin is encapsulated in micelles, preferably at least 97%, and more preferably at least 99%.

[0041] In another preferred example, the volume ratio of propylene glycol to D5W in the composition is about 6:100 to about 6:400, preferably about 6:150 to about 6:250, and more preferably about 6:200.

[0042] In another preferred embodiment, the particle size D50 of the micelles in the obtained Plinabulin composition for injection is 5-50 nm, preferably 10-30 nm.

[0043] In another preferred embodiment, the concentration of Plinabulin is about 0.08 mg / ml to about 0.4 mg / ml.

[0044] In another preferred embodiment, the volume ratio of polyethylene glycol 15-hydroxystearate and D5W in the liquid Plinabulin injection formulation is about 4:50 to about 4:500, preferably about 4:100 to about 4:500, more preferably about 4:100 to about 4:400, for example, about 4:100 to about 4:300, or about 4:150 to about 4:250.

[0045] In another preferred embodiment, in the liquid Plinabulin composition for injection, the weight ratio of 15-hydroxystearate polyethylene glycol to propylene glycol is 1:9, 2:8, 3:7, 4:6, 5:5, 6:4, 7:3, 8:2, or 9:1. In another preferred embodiment, the ratio of propylene glycol to 15-hydroxystearate polyethylene glycol is about 60:40 (wt:wt).

[0046] In another preferred embodiment, the total amount of impurities contained in the liquid Plinabulin composition for injection is less than 0.5%. In another preferred embodiment, the total amount of impurities contained in the liquid Plinabulin composition for injection is less than 0.1%.

[0047] In another preferred embodiment, the liquid Plinabulin composition for injection can be stably stored for about 8 hours to about 12 hours.

[0048] In another preferred embodiment, the liquid Plinabulin composition for injection can be stably stored at a temperature of 10° C. to about 37° C., preferably room temperature.

[0049] In another preferred embodiment, at least a portion of Plinabulin is encapsulated in micelles, preferably, at least 90% of Plinabulin is encapsulated in micelles.

[0050] In another preferred embodiment, greater than about 90% of the Plinabulin in the liquid injectable Plinabulin composition is encapsulated in micelles.

[0051] In another preferred embodiment, greater than about 93% of Plinabulin in the liquid Plinabulin composition for injection is encapsulated in micelles, preferably, greater than about 94% of Plinabulin is encapsulated in micelles, preferably, greater than about 95% of Plinabulin is encapsulated in micelles, preferably, greater than about 96% of Plinabulin is encapsulated in micelles, preferably, greater than about 97% of Plinabulin is encapsulated in micelles, preferably, greater than about 99% of Plinabulin is encapsulated in micelles.

[0052] In another preferred embodiment, the liquid Plinabulin composition for injection is prepared by the method described in the fourth aspect of the present invention.

[0053] A fourth aspect of the present invention provides a method for preparing a liquid injection composition of Plinabulin, wherein the method comprises:

[0054] Providing an initial liquid formulation comprising plinabulin, propylene glycol and polyethylene glycol 15-hydroxystearate; and

[0055] The initial liquid formulation was diluted in D5W at a dilution ratio of about 1:5 to about 1:50.

[0056] In another preferred embodiment, the initial liquid preparation is the Plinabulin micellar composition described in the second aspect of the present invention.

[0057] In another preferred example, the dilution ratio is about 1:10 to about 1:50, preferably, the dilution ratio is about 1:13 to about 1:30, and more preferably, the dilution ratio is about 1:20.

[0058] In another preferred embodiment, the initial liquid formulation comprises Plinabulin at a concentration of about 1 mg / ml to about 6 mg / ml, preferably comprises Plinabulin at a concentration of about 3 mg / ml to about 5 mg / ml, and more preferably comprises Plinabulin at a concentration of about 4 mg / ml.

[0059] In another preferred embodiment, the composition is stirred after diluting the initial liquid formulation in D5W, preferably for at least 5 minutes.

[0060] In another preferred embodiment, the concentration of Plinabulin in the liquid Plinabulin composition for injection is about 0.08 mg / ml to about 0.4 mg / ml.

[0061] In another preferred embodiment, the ratio of propylene glycol to 15-hydroxystearate polyethylene glycol in the liquid Plinabulin composition for injection is about 60:40 (wt:wt).

[0062] In another preferred embodiment, the initial liquid preparation is the Plinabulin micellar composition as described above.

[0063] In another preferred embodiment, the particle size of the micelles in the liquid Plinabulin composition for injection ranges from 90 to 200 nm, preferably from 100 to 150 nm, and more preferably from 100 to 110 nm.

[0064] The fifth aspect of the present invention provides a use of the Plinabulin micelle composition as described in the second aspect or the Plinabulin composition for liquid injection as described in the third aspect in the preparation of a drug for preventing and / or treating anti-tumor.

[0065] In another preferred embodiment, the tumor is selected from: lung cancer (such as small cell lung cancer, non-small cell lung cancer), prostate cancer, colon cancer, brain tumor (such as glioblastoma, glioblastoma multiforme, giant cell glioblastoma, metastatic brain tumor), head and neck cancer, gastric cancer, pancreatic cancer, breast cancer, kidney cancer, bladder cancer, ovarian cancer, cervical cancer, melanoma, myeloma, lymphoma or leukemia.

[0066] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features described in detail below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be listed here one by one. BRIEF DESCRIPTION OF THE DRAWINGS

[0067] FIG1 shows a Tyndall effect diagram of Plinabulin micellar composition.

[0068] FIG2 shows a diagram of the Tyndall effect of the liquid Plinabulin composition for injection. DETAILED DESCRIPTION

[0069] After extensive and in-depth research, the present inventors unexpectedly obtained a novel Plinabulin micelle composition with excellent stability. The Plinabulin micelle composition can improve the water solubility of Plinabulin and has excellent electrochemical stability, dilution stability and storage stability.

[0070] Specifically, the present invention unexpectedly prepares a Plinabulin micelle composition using Plinabulin monohydrate and a specific ratio of propylene glycol and 15-hydroxystearate polyethylene glycol, in which Plinabulin is encapsulated in micelles. The micelle composition has excellent stability, electrochemical stability, dilution stability and storage stability.

[0071] On the basis of obtaining the Plinabulin micelle composition, a D5W solution was added for dilution to obtain a Plinabulin composition for liquid injection. After dilution, the Plinabulin micelle composition still existed, and Plinabulin was encapsulated therein. The diluted injection had excellent instillation stability and storage stability, and the encapsulation rate of Plinabulin was high and the impurity content was low, and it can be used as an injection preparation of Plinabulin.

[0072] On this basis, the present invention was completed.

[0073] Disclosed herein are Plinabulin micellar encapsulated compositions and methods for preparing and using the Plinabulin compositions. Plinabulin ((3Z,6Z)-3-[(5-tert-butyl-1H-imidazol-4-yl)methylene]-6-(benzylidene)-2,5-piperazinedione) is a synthetic analog of the natural compound diketopiperazinephenylahistine. Plinabulin can be readily prepared according to the methods and procedures detailed in U.S. Patent Nos. 7,064,201 and 7,919,497, both of which are incorporated herein by reference in their entirety.

[0074] 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 the invention belongs. Although any methods and materials similar or equivalent to those described herein can also be used in the practice or testing of the present invention, the preferred methods and materials are described herein.

[0075] the term

[0076] 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 disclosure belongs. All patents, applications, published applications, and other publications are incorporated by reference in their entirety. If a term has multiple definitions herein, the definition in this section will prevail unless otherwise indicated.

[0077] Plinabulin as used herein includes crystalline forms or amorphous forms, such as Plinabulin monohydrate, Plinabulin anhydrate, and Plinabulin solvate, preferably monohydrate.

[0078] In the present invention, “injectable micellar formulation of Plinabulin”, “liquid injectable Plinabulin composition”, “liquid injectable formulation”, “liquid injectable Plinabulin formulation” and “liquid injectable Plinabulin composition” have the same meaning and can be used interchangeably.

[0079] The term "agent" is used herein to refer to a compound, a mixture of compounds, a biomacromolecule, or an extract made from biological material.

[0080] The terms "cancer," "neoplasm," and "carcinoma," used interchangeably herein, refer to cells that exhibit relatively autonomous growth, and thus exhibit an abnormal growth phenotype characterized by a significant loss of control over cell proliferation. Generally, cells of interest for detection or treatment herein include precancerous (e.g., benign), malignant, premetastatic, metastatic, and non-metastatic cells. Detection of cancer cells is particularly interesting.

[0081] As used herein, the term "subject" refers to a human or non-human mammal, such as a dog, cat, mouse, cow, sheep, pig, goat, non-human primate, or bird, such as a chicken, as well as any other vertebrate or invertebrate.

[0082] The term "mammal" is used in its ordinary biological sense. Thus, it specifically includes, but is not limited to, primates, including great apes (chimpanzees, apes, monkeys) and humans, cows, horses, sheep, goats, pigs, rabbits, dogs, cats, rodents, rats, mice, guinea pigs, or the like.

[0083] As used herein, the term "effective amount" or "therapeutically effective amount" refers to an amount of a therapeutic agent effective to alleviate to some extent one or more symptoms of a disease or condition, or to reduce the likelihood of its occurrence, and may include curing the disease or condition.

[0084] As used herein, the term "treat" refers to administering a compound or pharmaceutical composition to a subject for preventive and / or therapeutic purposes. The term "prophylactic treatment" refers to treating a subject who does not yet exhibit symptoms of a disease or condition, but who is susceptible to or otherwise at risk for a particular disease or condition, thereby reducing the likelihood that the patient will develop the disease or condition. The term "therapeutic treatment" refers to treating a subject who already has a disease or condition.

[0085] Formulations

[0086] In some embodiments, Plinabulin is provided in the form of a concentrated liquid formulation, which can then be diluted to prepare a diluted liquid injection formulation. In some embodiments, the concentrated liquid formulation includes one or more solvents, which may include 15-hydroxystearate polyethylene glycol ester and / or propylene glycol. In some embodiments, the one or more solvents include at least 30% (by weight) of 15-hydroxystearate polyethylene glycol ester, at least 35% (by weight) of 15-hydroxystearate polyethylene glycol ester, at least 40% (by weight) of 15-hydroxystearate polyethylene glycol ester, at least 45% (by weight) of 15-hydroxystearate polyethylene glycol ester, or a range including and / or spanning the above values. In some embodiments, the one or more solvents comprise at least 50% (by weight) propylene glycol, at least 55% (by weight) propylene glycol, at least 50% (by weight) propylene glycol, at least 55% (by weight) propylene glycol, at least 60% (by weight) propylene glycol, or a range including and / or spanning the foregoing values. In some embodiments, the one or more solvents comprise 40% (by weight) polyethylene glycol 15-hydroxystearate and 60% (by weight) propylene glycol. In some embodiments, the ratio of propylene glycol to polyethylene glycol 15-hydroxystearate in the liquid injectable formulation is approximately 60:40 (by weight).

[0087] In some embodiments, the concentrated liquid formulation is a micellar composition. Plinabulin is encapsulated in the micelles. In some embodiments, the ingredients of the micellar composition include polyethylene glycol 15-hydroxystearate, propylene glycol, and Plinabulin, wherein the Plinabulin is encapsulated in the micelles. In some embodiments, at least 60%, at least 62%, at least 64%, at least 66%, at least 68%, at least 70%, at least 72%, at least 74%, at least 76%, at least 78%, at least 80%, at least 82%, at least 84%, at least 86%, at least 88%, at least 90%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or a range including and / or spanning the foregoing values, of the Plinabulin encapsulated in the micelles.

[0088] In some embodiments, the Plinabulin in the micellar composition is Plinabulin monohydrate. When the anhydrous form of Plinabulin is used, the resulting micellar composition has poor solubility.

[0089] The concentration of Plinabulin in the concentrated liquid formulation may be between 1 mg / ml and 10 mg / ml, between 2 mg / ml and 6 mg / ml, or about 4 mg / ml.

[0090] In some embodiments, the concentrated liquid formulation can be diluted with a diluent to form a diluted liquid injection formulation. In some embodiments, the diluent is water. In some embodiments, the diluent is saline. In some embodiments, the diluent is a glucose aqueous solution. The concentration of glucose can be 1% to 20%, 2% to 10%, or about 5% (i.e., D5W).

[0091] In some embodiments, the diluted liquid injection formulation comprises polyethylene glycol 15-hydroxystearate, propylene glycol and D5W (i.e., glucose and water). In some embodiments, the volume ratio of propylene glycol to D5W in the liquid injection formulation of prinab is in the range of about 6:50 to about 6:500, about 6:50 to about 6:450, about 6:50 to about 6:400, about 6:50 to about 6:350, about 6:50 to about 6:300, about 6:50 to about 6:250, about 6:50 to about 6:200, about 6:50 to about 6:150, about 6:50 to about 6:100, about 6:100 to about 6:50, about 6:100 to 6:400, about 6:150 to about 6:250, or about 6:200. In some embodiments, the volume ratio of propylene glycol to D5W in the liquid Plinabulin injection formulation is about 6:70. In some embodiments, the volume ratio of propylene glycol to D5W in the liquid Plinabulin injection formulation is about 6:140. In some embodiments, the volume ratio of propylene glycol to D5W in the liquid Plinabulin injection formulation is about 6:400.

[0092] In some embodiments, the volume ratio of polyethylene glycol 15-hydroxystearate and D5W in the liquid Plinabulin injection formulation is in the range of about 4:50 to about 4:500, about 4:100 to about 4:500, about 4:100 to about 4:400, about 4:100 to about 4:300, about 4:150 to about 4:250, or about 4:200.

[0093] In some embodiments, the Plinabulin in the liquid Plinabulin injection formulation (composition) is Plinabulin monohydrate.

[0094] In some embodiments, the concentration of Plinabulin in the liquid injectable Plinabulin formulation (composition) is about 0.02 mg / ml, 0.03 mg / ml, 0.04 mg / ml, 0.05 mg / ml, 0.06 mg / ml, 0.07 mg / ml, 0.08 mg / ml, 0.09 mg / ml, 0.1 mg / ml, 0.11 mg / ml, 0.12 mg / ml, 0.13 mg / ml, 0.14 mg / ml, 0.15 mg / ml, 0.16 mg / ml, 0.17 mg / ml, 0.18 mg / ml, 0.19 mg / ml, 0.2 mg / ml, 0.3 mg / ml, 0.4 mg / ml, 0.5 mg / ml, 1 mg / ml, 2 mg / ml, 3 mg / ml, 4 mg / ml, or a range including and / or spanning the above values. In some embodiments, the concentration of Plinabulin in the liquid composition for injection is about 0.08 mg / ml to about 0.4 mg / ml. In some embodiments, the concentration of Plinabulin in the liquid composition for injection is about 0.1 mg / ml to about 0.3 mg / ml. In some embodiments, the concentration of Plinabulin in the liquid composition for injection is about 0.2 mg / ml.

[0095] In some embodiments, the liquid Plinabulin formulation (composition) for injection contains less than 5%, less than 4%, less than 3%, less than 2%, less than 1%, less than 0.5% impurities, or a range including and / or spanning the above values. In some embodiments, the liquid Plinabulin formulation (composition) for injection contains less than 0.5% ether impurities. In some embodiments, the liquid Plinabulin formulation (composition) for injection contains less than 0.5% alcohol impurities. In some embodiments, the liquid Plinabulin formulation (composition) for injection contains less than 0.5% water.

[0096] In some embodiments, the liquid injectable Plinabulin formulation (composition) can be stored stably for about 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours, 24 hours, 36 hours, 48 ​​hours, or a range including and / or spanning the above values.

[0097] In some embodiments, the liquid Plinabulin injection formulation (composition) can be stably stored at 10°C, 12°C, 14°C, 16°C, 18°C, 20°C, 22°C, 24°C, 26°C, 28°C, 30°C, 32°C, 34°C, 36°C, 38°C, 40°C, or a range including and / or spanning the above values.

[0098] In some embodiments, when the concentrated liquid formulation is diluted with a diluent such as D5W, a portion of the Plinabulin is encapsulated within micelles. In some embodiments, the micelles are formed by the interaction between polyethylene glycol 15-hydroxystearate and D5W. In some embodiments, at least 60%, at least 62%, at least 64%, at least 66%, at least 68%, at least 70%, at least 72%, at least 74%, at least 76%, at least 78%, at least 80%, at least 82%, at least 84%, at least 86%, at least 88%, at least 90%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or a range including and / or spanning the foregoing values, of the Plinabulin in the injectable liquid formulation (composition) is encapsulated within micelles.

[0099] In some embodiments, the average encapsulation efficiency of the liquid formulation is at least 90%, at least 92%, at least 94%, at least 96%, at least 98%, or a range including and / or spanning the aforementioned values.

[0100] Preparation method

[0101] In certain aspects, methods of preparing the liquid injectable formulations (compositions) described herein are disclosed. In some embodiments, the method of preparing a liquid injectable Plinabulin composition comprises providing an initial concentrated liquid formulation comprising Plinabulin, propylene glycol, and polyethylene glycol 15-hydroxystearates; and diluting the initial liquid formulation in D5W. In various embodiments, the dilution ratio between the initial concentrated liquid formulation and D5W is from about 1:5 to about 1:50, from about 1:10 to about 1:50, from about 1:10 to about 1:40, from about 1:15 to about 1:30, from about 1:15 to about 1:25, or from about 1:20. In some embodiments, the dilution ratio is from about 1:10 to about 1:400. In some embodiments, the dilution ratio is from about 1:10 to about 1:300. In some embodiments, the dilution ratio is from about 1:10 to about 1:200. In some embodiments, the dilution ratio is from about 1:10 to about 1:100. In some embodiments, the dilution ratio is from about 1:10 to about 1:80. In some embodiments, the dilution ratio is from about 1:10 to about 1:60. In some embodiments, the dilution ratio is from about 1:10 to about 1:50. In some embodiments, the dilution ratio is from about 1:10 to about 1:40. In some embodiments, the dilution ratio is from about 1:10 to about 1:30. In some embodiments, the dilution ratio is from about 1:10 to about 1:20. In some embodiments, the dilution ratio is from about 1:20 to about 1:400. In some embodiments, the dilution ratio is from about 1:20 to about 1:300. In some embodiments, the dilution ratio is from about 1:20 to about 1:200. In some embodiments, the dilution ratio is from about 1:20 to about 1:100. In some embodiments, the dilution ratio is from about 1:20 to about 1:80. In some embodiments, the dilution ratio is from about 1:20 to about 1:60. In some embodiments, the dilution ratio is from about 1:20 to about 1:50. In some embodiments, the dilution ratio is about 1:20 to about 1:40. In some embodiments, the dilution ratio is about 1:20 to about 1:30.

[0102] In some embodiments, the dilution method includes introducing the initial concentrated liquid formulation into (e.g., by injection) a container containing a certain volume of D5W. In some embodiments, the container is an intravenous (IV) bag. In some embodiments, after the initial concentrated liquid formulation is introduced into the container, the mixture is stirred. Various stirring methods include hand shaking, hand stirring or vortexing. In various embodiments, the mixture is stirred for at least 1 minute, at least 2 minutes, at least 3 minutes, at least 5 minutes, at least 10 minutes, at least 20 minutes, at least 30 minutes, at least 45 minutes, at least 60 minutes, at least 90 minutes, or include and / or span the range of the above values.

[0103] In some embodiments of preparing the initial concentrated liquid formulation, propylene glycol is added first, followed by polyethylene glycol 15-hydroxystearate, and then Plinabulin. In some embodiments, polyethylene glycol 15-hydroxystearate is added first, followed by propylene glycol, and then Plinabulin. In some embodiments, the initial concentrated liquid formulation is mixed at room temperature.

[0104] In some embodiments of preparing the initial concentrated liquid formulation, propylene glycol is heated to about 25° C. and maintained at about 25° C. during the addition of polyethylene glycol 15-hydroxystearate. In some embodiments, plinabulin is added to polyethylene glycol 15-hydroxystearate and propylene glycol and mixed at about 25° C.

[0105] In some embodiments of preparing the initial concentrated liquid formulation, propylene glycol is heated to about 40° C. and maintained at about 40° C. during the addition of polyethylene glycol 15-hydroxystearate. In some embodiments, Plinabulin is added to polyethylene glycol 15-hydroxystearate and propylene glycol and mixed at about 40° C.

[0106] In some embodiments of preparing the initial concentrated liquid formulation, polyethylene glycol 15-hydroxystearate is heated to about 25° C. and maintained at about 25° C. while propylene glycol is added. In some embodiments, plinabulin is added to polyethylene glycol 15-hydroxystearate and propylene glycol and mixed at about 25° C.

[0107] In some embodiments of preparing the initial concentrated liquid formulation, polyethylene glycol 15-hydroxystearate is heated to about 40° C. and maintained at about 40° C. while propylene glycol is added. In some embodiments, plinabulin is added to polyethylene glycol 15-hydroxystearate and propylene glycol and mixed at about 40° C.

[0108] In some embodiments of preparing the initial concentrated liquid formulation, polyethylene glycol 15-hydroxystearate is heated to about 60° C. and maintained at about 60° C. while propylene glycol is added. In some embodiments, plinabulin is added to polyethylene glycol 15-hydroxystearate and propylene glycol and mixed at about 60° C.

[0109] Uses and methods

[0110] Some embodiments relate to a method of preventing or reversing cancer progression in a subject. In some embodiments, the method comprises administering to the subject an injectable liquid formulation described herein. Some embodiments relate to a method of inhibiting cancer progression. Further uses of the injectable liquid formulations described herein include U.S. Patent Nos. 7,919,497; 10,238,650; 10,155,748; 10,076,518; and 10,596,169; and PCT Publication Nos. WO 2016 / 130839; WO 2017 / 214052; WO 2018 / 144764; WO 2018 / 169887; WO 2019 / 147615; WO 2019 / 152530; WO 2020 / 037285; WO 2021 / 076485; and WO 2021 / 225908; all of which are incorporated herein by reference in their entirety.

[0111] In some embodiments, the treatment regimen includes administering the injectable liquid formulation described herein once every 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks or 8 weeks. In some embodiments, the treatment regimen includes administering the injectable liquid formulation described herein 2 times every 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks or 8 weeks. In some embodiments, the treatment regimen includes administering the injectable liquid formulation described herein once every 1 week within a treatment cycle of 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks or 8 weeks. In some embodiments, the treatment regimen includes administering the injectable liquid formulation described herein twice every 1 week within a treatment cycle of 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks or 8 weeks. In some embodiments, the treatment regimen includes administering the injectable liquid formulation described herein twice every 1 week within a treatment cycle of 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks or 8 weeks. In some embodiments, the treatment regimen includes administering the injectable liquid formulation described herein on the 1st day, the 8th day and the 15th day in a 21-day treatment cycle.

[0112] As long as the regimen is clinically tolerable, the treatment cycle can be repeated. In some embodiments, the treatment cycle of the injectable liquid formulation described herein is repeated n times, wherein n is an integer in the range of 2 to 30. In some embodiments, n is 2, 3, 4, 5, 6, 7, 8, 9 or 10. In some embodiments, a new treatment cycle can occur immediately after the previous treatment cycle is completed. In some embodiments, a new treatment cycle can occur within a period of time after the previous treatment cycle is completed. In some embodiments, a new treatment cycle can occur 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks or 7 weeks after the previous treatment cycle is completed.

[0113] In some embodiments, the method comprises administering at about 5 mg / m 2 Up to 150 mg / m 2 In some embodiments, the dosage of Plinabulin is greater than 20 mg / m 2 In some embodiments, the dosage of Plinabulin is greater than 30 mg / m 2 In some embodiments, the dosage of Plinabulin is greater than 40 mg / m 2 .

[0114] In some embodiments, the injectable liquid formulations described herein are administered on day 1 of a 14-day dosing cycle. In some embodiments, the injectable liquid formulations described herein are administered on day 1 of a 21-day dosing cycle.

[0115] In some embodiments, the injectable liquid formulations described herein are co-administered with one or more G-CSF drugs.

[0116] Some embodiments include kits comprising one or more containers.

[0117] In some embodiments, the container comprises plastic or glass, or a combination thereof, including but not limited to any one or more plastics or glasses used by one of skill in the art in light of the teachings herein.

[0118] In some embodiments, the container is a vial. In some embodiments, the vial contains about 1 mg, 2 mg, 5 mg, 10 mg, 20 mg, 30 mg, 40 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, 190 mg, 200 mg of Plinabulin, or an amount including and / or spanning a range of the above values. In some embodiments, the vial contains Plinabulin, propylene glycol, and PEG15-hydroxystearates. In some embodiments, the vial contains about 4 mg / ml of Plinabulin, propylene glycol, and PEG15-hydroxystearates in a ratio of about 60:40 (wt:wt). In some embodiments, the volume of the liquid formulation in the vial is about 10 ml.

[0119] In some embodiments, the container is an IV bag. In some embodiments, the IV bag comprises D5W. In some embodiments, the volume of D5W in the IV bag is about 50 ml to about 500 ml, about 100 ml to about 500 ml, about 100 ml to about 400 ml, about 100 ml to about 300 ml, about 150 ml to about 250 ml, or about 200 ml.

[0120] In some embodiments, the kit comprises a vial as described above and an intravenous syringe as described above.

[0121] Preparation of Plinabulin micellar composition

[0122] After melting 15-hydroxystearate polyethylene glycol ester at 50℃±5℃, weigh the prescribed amount of melted 15-hydroxystearate polyethylene glycol ester and add it to a nitrogen-filled mixing tank, then add the prescribed amount of propylene glycol, maintain the temperature at 40℃±5℃, and stir for 30 minutes.

[0123] An appropriate amount of a mixture of 15-hydroxystearic acid polyethylene glycol ester and propylene glycol was transferred from the DGJ-14 batching tank to a beaker and a Duran bottle for later use. The prescribed amount of Plinabulin was weighed into an isolator in the cytotoxic drug weighing room. The 15-hydroxystearic acid polyethylene glycol ester and propylene glycol mixture in the beaker was added to the mixture, stirred for 5 minutes, and then transferred to the batching tank. The beaker was thoroughly rinsed with the 15-hydroxystearic acid polyethylene glycol ester and propylene glycol mixture from the Duran bottle three times before being transferred to the batching tank. After stirring for 1 hour using a magnetic stirrer (40 Hz), a sample was taken for intermediate product testing (the sample should be protected from light). The temperature of the drug solution was maintained at 40°C ± 5°C throughout the entire process. The intermediate product was sampled and tested according to the sampling plan for properties, moisture content, density, related substances, microbial limits, bacterial endotoxins, and content. The time from preparation to sterilization and filtration should not exceed 6 hours. A Tyndall effect diagram of the Plinabulin micellar composition is shown in Figure 1.

[0124] Plinabulin composition for injection

[0125] The Plinabulin micelle composition described above is diluted in D5W at a dilution ratio of about 1:5 to about 1:50 (wt) to obtain a Plinabulin composition for injection, wherein the Tyndall effect diagram for a dilution ratio of 1:20 is shown in Figure 2.

[0126] Compared with the existing technology, this application has the following main advantages

[0127] 1. The Plinabulin micellar composition of the present application has excellent dilution stability.

[0128] 2. The Plinabulin micelle composition of the present application, and in particular the prepared Plinabulin composition for injection, have excellent storage stability.

[0129] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are intended to illustrate the present invention only and are not intended to limit the scope of the invention. The experimental methods in the following examples, for which specific conditions are not specified, are generally based on conventional conditions or the conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are calculated by weight.

[0130] Example 1 Effects of different orders of adding the formula on the dissolution rate of Plinabulin and the particle size distribution of the final product obtained.

[0131] 1. Different order of adding formula (propylene glycol / 15-hydroxystearate polyethylene glycol ratio: 6 / 4)

[0132] Table 1-1 Composition

[0133] 1.1 Add propylene glycol first, then add 15-hydroxystearate polyethylene glycol, and finally add the medicine

[0134] Plinabulin micellar compositions A, B, and C were prepared by adding 15-hydroxystearate polyethylene glycol to ethylene glycol at 25°C (solid state), 40°C (molten state), and 60°C, followed by the addition of Plinabulin. These compositions were then diluted 1:20 with 5% glucose injection to produce liquid injectable Plinabulin compositions (preparations). Particle size analysis revealed that under the preparation conditions where propylene glycol was added first, followed by 15-hydroxystearate polyethylene glycol, and finally the drug, the dissolution rate of the API accelerated with increasing temperature. At 25°C, the drug required stirring for nearly 2 hours to dissolve, while at 40°C and 60°C, the API completely dissolved in approximately 10 minutes. The average particle size of Plinabulin micellar compositions A and B was approximately 100 nm. Particle size analysis of Plinabulin micellar composition C was not possible. The particle size distributions of the different Plinabulin micellar compositions after a 1:20 dilution were similar, indicating that the preparation temperature had little effect on the particle size distribution after dilution.

[0135] 1.2 Add 15-hydroxystearate polyethylene glycol first, then add propylene glycol and finally add the medicine

[0136] Plinabulin micellar compositions D, E, and F were prepared by adding ethylene glycol to 15-hydroxystearate polyethylene glycol at 25°C (solid state), 40°C (molten state), and 60°C, followed by plinabulin. These compositions were then diluted with 5% glucose injection at a ratio of 1:20 to produce liquid injectable plinabulin compositions (preparations). Particle size analysis revealed that when 15-hydroxystearate polyethylene glycol was added first, followed by propylene glycol, the drug dissolution rate increased with increasing temperature. At 25°C, the drug remained undissolved after 5 hours of stirring. However, at 40°C and 60°C, the drug completely dissolved within 15 minutes. Particle size distribution analysis revealed that the preparation temperature had little effect on the particle size distribution of the micellar compositions, and that the order of adding 15-hydroxystearate polyethylene glycol and propylene glycol did not affect the particle size of the preparations.

[0137] 1.3 Add the medicine first, then add propylene glycol and finally add 15-hydroxystearate polyethylene glycol

[0138] The API was first added, followed by propylene glycol, and then 15-hydroxystearate polyethylene glycol ester at 25°C (solid), 40°C (molten), and 60°C, respectively, to prepare Plinabulin micellar compositions G, H, and I. These compositions were then diluted with 5% glucose injection at a ratio of 1:20 to obtain liquid Plinabulin compositions (preparations) for injection. Particle size measurements showed that at temperatures above 40°C, the API dissolved in the propylene glycol solvent relatively quickly, and the solution remained clear after the addition of 15-hydroxystearate polyethylene glycol ester. However, when the propylene glycol temperature was 25°C, the API was difficult to dissolve. Particle size distribution measurements showed that dissolving the drug in propylene glycol first had no effect on the particle size distribution.

[0139] 1.4 Add the medicine first and then add the mixture of propylene glycol and 15-hydroxystearate polyethylene glycol

[0140] The API was first added, followed by a pre-mixed solution of propylene glycol and 15-hydroxystearate polyethylene glycol, each at 40°C, to prepare Plinabulin micellar composition J. This was then diluted with 5% glucose injection at a ratio of 1:20 to obtain a liquid Plinabulin composition (preparation) for injection. Particle size measurement revealed that the average particle size of the preparation containing the API first and then the propylene glycol and 15-hydroxystearate mixture increased from 104.61 nm to 112.52 nm, and D10 significantly increased, compared to the micellar composition of 1.1. However, there was no significant difference in the particle size of the diluted sample.

[0141] 1.5 Add the medicine first, then add 15-hydroxystearate polyethylene glycol, and finally add propylene glycol

[0142] The API was first added, followed by 15-hydroxystearic acid polyethylene glycol ester melted at 40°C, the temperature was maintained at 40°C, stirring was continued, and propylene glycol was added to prepare Plinabulin micellar composition H, which was then diluted with 5% glucose injection at a ratio of 1:20 to obtain a Plinabulin composition (preparation) for injection. Particle size measurement showed that the API was insoluble in 15-hydroxystearic acid polyethylene glycol ester. After adding propylene glycol, the dissolution time of the API was increased compared to 1.1. The particle size results showed that compared with the particle size results in 1.1, the particle size of the stock solution was significantly increased when the API was first dispersed in 15-hydroxystearic acid polyethylene glycol ester and then propylene glycol was added. The average particle size of the micelle composition increased from 104.61 nm to 127.06 nm, with no significant change in D10, D50 from 100.08 nm to 118.03 nm, and D90 from 145.88 nm to 192.41 nm. However, there was no significant difference in the particle size of the 1:20 diluted sample.

[0143] In summary, the preparation time of the micellar composition decreases with increasing preparation temperature. Regarding the process flow, when the API is added last, the order of adding 15-hydroxystearate polyethylene glycol and propylene glycol has no significant effect on the preparation process or the particle size of the micellar composition. When the raw material is added first, dispersing with propylene glycol first and then adding 15-hydroxystearate polyethylene glycol shortens the preparation time, and the particle size results are not significantly different from the production conditions; when a mixture of 15-hydroxystearate polyethylene glycol and propylene glycol is added, the preparation time and phenomena are not significantly different, but the particle size distribution results of the micelle composition show that the average particle size, D10, D50, and D90 all increase; dispersing with 15-hydroxystearate polyethylene glycol first and then adding propylene glycol increases the preparation time slightly, and the particle size distribution results of the micelle composition show that the average particle size, D10, D50, and D90 all increase, and are larger than the sample with the mixture of 15-hydroxystearate polyethylene glycol and propylene glycol added.

[0144] 2 Different prescription ratios and temperatures

[0145] 2.1 Propylene glycol: 15-hydroxystearate polyethylene glycol (7:3)

[0146] Table 1-2 Composition

[0147] Plinabulin micellar compositions K and L were prepared by adding 40°C (molten) and 60°C 15-hydroxystearate polyethylene glycol to ethylene glycol, respectively, followed by the addition of Plinabulin. These compositions were then diluted with 5% glucose injection at a ratio of 1:20 to produce liquid injectable Plinabulin compositions (preparations). Particle size analysis revealed that at a 7:3 ratio of propylene glycol to 15-hydroxystearate polyethylene glycol, the solution was turbid. Although the API dissolved after addition, the solution remained turbid. The API dissolution rate was positively correlated with temperature, and the clarity of the sample solution increased with increasing temperature. Due to the turbidity and non-homogeneous nature of the sample, particle size analysis was difficult. Particle size analysis of the diluted sample of the preparation prepared at 60°C alone yielded similar particle size and distribution to those of the preparation prepared in step 1.1.

[0148] 2.2 Propylene glycol: 15-hydroxystearate polyethylene glycol (3:7)

[0149] Table 1-3 Composition

[0150] Plinabulin micellar compositions M and N were prepared by adding 15-hydroxystearate polyethylene glycol at 40°C (molten) and 60°C (molten) to ethylene glycol, followed by Plinabulin. These compositions were then diluted with 5% glucose injection at a ratio of 1:20 to produce liquid injectable Plinabulin compositions (preparations). Particle size analysis revealed that the dissolution rate of the API increased with increasing temperature. Particle size distribution results showed that changing the ratio of propylene glycol to 15-hydroxystearate polyethylene glycol to 3:7 significantly reduced the particle size of the micellar compositions.

[0151] In summary, changing the ratio of propylene glycol to polyethylene glycol 15-hydroxystearate (PEG-15HST) resulted in changes in the properties of the mixed solution. When the ratio of propylene glycol to polyethylene glycol 15-hydroxystearate was 7:3, the solution became turbid and significantly affected the drug dissolution rate. When the ratio of propylene glycol to polyethylene glycol 15-hydroxystearate was 3:7, the preparation process remained unchanged, but the resulting particle size was significantly reduced. It is possible that within a certain range, the micelle size gradually decreases with increasing surfactant concentration.

[0152] Example 2

[0153] This example studies the micelle properties of the prepared injectable Plinabulin solution. The encapsulation efficiency of the diluted Plinabulin was determined by separating the drug encapsulated from the free drug in the micelles.

[0154] Because the target compound to be determined was Plinabulin in the formulation, a liquid chromatography-mass spectrometry method was selected for this product, and the reproducibility and specificity of the method were confirmed. The liquid chromatography-mass spectrometry conditions and experimental procedures are shown in Table 2-1.

[0155] Table 2-1 Liquid phase determination method - specific parameters

[0156] Flow rate: 1.2 ml / min; column temperature: 40°C; syringe temperature: 20°C; detection wavelength: 330 nm; injection volume: 10 μl

[0157] Samples of Plinabulin solution were prepared according to the assay method in Table 2-1. The reproducibility of the method was determined.

[0158] Table 2-2 Solution preparation process

[0159] Inject 10 μL of the Plinabulin solution, reference solution, and test solution into the liquid chromatograph. Calculate the peak areas and determine the Plinabulin content in the product according to the method standard. The measurement results are shown in Tables 2-3 and 2-4.

[0160] Table 2-3 Detection Method Validation - Reproducibility - System Suitability

[0161] Table 2-4 Validation of the assay method - Reproducibility - Content determination

[0162] It can be seen from the test data and chromatogram that the determination method of this product has good sensitivity and reproducibility, and can meet the needs of determining the encapsulation rate of the diluted preparation of this product.

[0163] The equilibrium solubility of the drug in the solvent system of the injection preparation without a solubilizer (15-hydroxystearate polyethylene glycol) was determined to provide a reference for the determination of free drug. 40 mg of Plinabulin API was added to 6.0 g of propylene glycol and then added to 200 ml of D5W. The resulting solution was continuously shaken at 25°C and 100 rpm for 48 hours. The test solution was spun down by centrifugation and filtered through a 0.45 μm nylon filter. The supernatant and subsequent filtrate were injected into the HPLC system and the peak areas were recorded. The results are shown in Table 2-5 below.

[0164] Table 2-5 Solution Treatment Studies - Centrifugation and Nylon Filtration

[0165] The experimental data demonstrates that Plinabulin is significantly adsorbed by the 0.45 μm nylon membrane filter. Diluting the centrifuged supernatant with acetonitrile significantly increases the peak area for Plinabulin, indicating that undissolved Plinabulin remains in the supernatant. Therefore, direct centrifugation should not be used to prepare the test solution for Plinabulin; instead, a suitable membrane filter should be selected for filtration.

[0166] After 52 hours of shaking, the test solution was collected, filtered through a 0.45 μm glass fiber membrane, and diluted. The subsequent filtrate was then analyzed with acetonitrile to confirm the dissolution of Plinabulin. The corresponding continuous filtrate was injected into a high-performance liquid chromatography system, and the peak areas were recorded. The results are shown in Table 2-6.

[0167] Table 2-6 Solution Treatment Research - Glass Fiber Filtration

[0168] From the test results, it can be seen that the raw material solution of plibulin was directly filtered through a 0.45 μm glass fiber membrane, and the peak area after filtration was multiplied by acetonitrile for dilution.

[0169] Equilibrium solubility

[0170] The corresponding propylene glycol-D5W injection was prepared. The preparation process is as follows:

[0171] Diluent 1 (corresponding to a dilution ratio of 1:20). Weigh approximately 6 g of propylene glycol into a beaker, add 200 ml of D5W injection, and stir evenly.

[0172] Diluent 2 (corresponding to a dilution ratio of 1:30): Weigh approximately 4 g of propylene glycol, place it in a beaker, add 200 ml of D5W injection, and mix well.

[0173] Diluent 3 (corresponding to a dilution ratio of 1:50): Weigh approximately 2.4 g of propylene glycol, place it in a beaker, add 200 ml of D5W injection, and mix well.

[0174] The scheme for equilibrium solubility study is shown in Table 2-7.

[0175] Table 2-7 Equilibrium solubility study plan

[0176] Acceptance criteria: The concentration of plinabulin in each solution was determined until there was no significant difference between two consecutive determinations, i.e., equilibrium solubility.

[0177] In the content determination method of this product, the reference solution is prepared with absolute ethanol. Since this product is used to determine plasma proteins in D5W solution, the effects of different diluents on the determination were investigated. The experimental design is as follows, see Table 2-8.

[0178] Table 2-8 Confirmation of control dilution solvent

[0179] 10 μl of each reference solution prepared with different diluents and 48-hour test solution were injected into the HPLC system, and the solubility of Plinabulin in the solution was calculated using the external standard method. The results are shown in Table 2-9.

[0180] Table 2-9 Different control dilution solvents-solubility results

[0181] The results showed no significant differences in the results of the Plinabulin reference solution prepared with anhydrous ethanol and the diluent (propylene glycol / D5W injection) in each diluent. Therefore, in the subsequent encapsulation efficiency determination method, the Plinabulin reference solution was prepared with absolute ethanol.

[0182] At different time points, the test sample solutions were filtered through a 0.45 μm glass fiber membrane and injected into a high-performance liquid chromatography system. The equilibrium solubility of Plinabulin in the solution at different ratios at each time point was calculated using the external standard method. The results are shown in Table 2-10.

[0183] Table 2-10 Equilibrium solubility of Plinabulin at different dilution levels

[0184] Encapsulation rate of diluted Plinabulin for injection

[0185] Based on the above research, the final method for determining packaging efficiency is as follows:

[0186] (1:10) and (1:20) dilution test solution: Take prinabulin concentrate (4 mg / ml prinabulin in propylene glycol / polyoxyethylene 15-hydroxystearate, 60:40 (weight ratio)), place it in a 250ml measuring bottle, add 100ml (corresponding to a 1:10 dilution) or 200ml (corresponding to a 1:20 dilution) of D5W injection, and shake up and down 30 times for 1 minute.

[0187] (1:30) and (1:50) dilution test solutions: Take prinabulin concentrate for injection (4 mg / ml prinabulin in propylene glycol / polyoxyethylene 15-hydroxystearate, 60:40 (weight ratio)), place it in a 500ml volumetric flask, add 300ml (corresponding to a 1:30 dilution) or 500ml (corresponding to a 1:50 dilution) of D5W injection, and shake up and down 30 times for 1 minute.

[0188] Free test solution. Take approximately 4 mL of each dilution of the test solution and place it in an ultrafiltration centrifuge tube containing 30 kD regenerated cellulose. Centrifuge at 4500 g, discarding the ultrafiltration tube every 10 minutes. Add approximately 4 mL of the test solution to all solutions and continue centrifugation. Repeat this process six times (for a total of 60 minutes). Use the filtrate after 60 minutes as the free test solution.

[0189] Prepare the total amount of test solution: Take an appropriate amount of each diluted test solution and dilute it with absolute ethanol to prepare a solution containing approximately 2.5 μg of plinabulin per 1 ml as the total amount of test solution.

[0190] Stock solution of reference substance: Take about 25 mg of Plinabulin reference substance, accurately weigh it, put it into a 100 ml volumetric flask, add absolute ethanol to dissolve it and dilute it to the mark, and shake well.

[0191] Preparation of reference solution: Accurately weigh 1 ml of the above reference solution stock solution and place it in a 100 ml volumetric flask. Dilute to the mark with absolute ethanol and shake well to prepare the reference solution.

[0192] Sensitivity Solution: Accurately measure an appropriate amount of the above reference solution and dilute it with absolute ethanol to prepare a solution containing approximately 0.05 μg of Plinabulin per 1 ml.

[0193] Chromatographic conditions: Octadecylsilane bonded silica gel as the filler (ACE C18 4.6 mm × 150 mm, 5.0 μm or equivalent chromatographic column); 0.01 mol / L phosphate buffer (0.82 g ± 0.01 g of sodium dihydrogen phosphate monohydrate and 1.20 g ± 0.01 g of sodium phosphate, dissolved in water to dilute to 1000 ml, and shaken well) as mobile phase A, acetonitrile as mobile phase B, gradient elution according to the table below; flow rate 1.2 ml per minute; column temperature 40°C; injector temperature 30°C; detection wavelength 330 nm; injection volume 20 μl (see Table 2-11).

[0194] Table 2-11

[0195] System suitability requirements: The relative deviation of the Plinabulin peak area should not exceed 2.0% when the reference solution is injected five times continuously; the signal-to-noise ratio of the Plinabulin peak height in the chromatogram of the sensitivity solution should be greater than 10.

[0196] Determination method: Accurately determine the total amount of reference solution, test solution and free test solution, inject into the liquid chromatography system, record the chromatogram, and calculate the peak area according to the external standard method. The calculation method is as follows:

[0197] C 对 : Reference substance concentration (μg / ml).

[0198] A 对 : Peak area of ​​reference substance.

[0199] C 游离 : Free concentration of test sample (μg / ml).

[0200] A 游离 : Free peak area of ​​test sample.

[0201] C 总量 : Total concentration of test sample (μg / ml).

[0202] A 总量 : Peak area of ​​the total amount of test sample.

[0203] f: dilution multiple of the total amount of the test sample.

[0204] Using the above method, the encapsulation rate of each dilution level of 6 batches of samples was measured, and the measurement results are as follows:

[0205] Table 2-12 Encapsulation rate results of each dilution level of each batch of samples

[0206] The results showed that the encapsulation efficiency of Plinabulin micelles decreased with the increase of dilution ratio.

[0207] Effect of stirring method on encapsulation rate

[0208] The effects of different shaking times and methods on micelle properties were investigated. Using samples from batch 004A, concentrated Plinabulin (4 mg / mL in propylene glycol / polyethylene glycol 15-hydroxystearate) was diluted with D5W to dilution ratios of 1:10, 1:20, 1:30, and 1:50. Mixtures were prepared using either manual shaking at varying frequencies or vortexing for varying durations, and encapsulation efficiency was determined. The stirring method was as follows:

[0209] Table 2-13 Shaking method research - specific parameters

[0210] The encapsulation rate results are as follows:

[0211] Table 2-14 Study on the shaking method - Encapsulation rate results of solutions at different dilution levels

[0212] The results showed that the encapsulation efficiency of each sample ranged from 95% to 99% for samples prepared using four methods: manual shaking 30 times per minute, manual shaking 90 times per minute, stirring for 1 minute, and stirring for 3 minutes. This range indicates that diluting the sample using different shaking methods has little effect on the encapsulation efficiency. Therefore, it can be inferred that the encapsulation efficiency of sample solutions diluted by different personnel is relatively stable during clinical use.

[0213] Study on the dilution resistance of diluted preparations in PBS solution

[0214] Batch samples were diluted (at 1:10, 1:20, 1:30, and 1:50) to simulate a clinical infusion process. The diluted test solution was infused into 5 L of pH 7.4 phosphate buffer over approximately 30 minutes. Based on human blood volume (approximately 7% to 8% of body weight, or approximately 4.2 to 4.8 L for a 60 kg body weight), 5 L of pH 7.4 phosphate buffer was used to simulate human blood volume. Sampling points were at 10 minutes, 20 minutes, and completion of the infusion (except for the case where the test solution was diluted at a 1:50 ratio, in which the infusion was completed over approximately 60 minutes, in which case sampling points were at 20 minutes, 40 minutes, and completion of the infusion).

[0215] Since the drug concentration in the dilution resistance test was low (total concentration was in the range of 7 μg / ml to 8 μg / ml) and it was a phosphate buffer system, it was determined that the regenerated cellulose filter membrane with a cutoff molecular weight of 30 kd had filtration and adsorption effects at this concentration level.

[0216] (1:20) dilution level solution: Accurately weigh approximately 10.36 g of plasma concentrate into a 250 ml volumetric flask, add 200 ml of D5W injection, and shake up and down 30 times within 1 minute to obtain the solution.

[0217] Free test solution: Measure 2 ml of the diluted solution above and place it in 50 ml of pH 7.4 phosphate buffer (37°C). Stir for 1 minute. Take approximately 4 ml of the solution (reconstitute the solution and replace the solution in the centrifuge tube every 10 minutes) and place it in an ultrafiltration centrifuge tube (regenerated cellulose molecular weight 30 kd). Centrifuge (4500g) for 20 min, 40 min, 50 min, 60 min, 70 min, and 80 min. The filtrate is used as the free test solution.

[0218] Prepare the total amount of test solution: accurately measure 2 ml of the above solution, place it in a 50 ml volumetric flask, dilute to the mark with absolute ethanol, shake well, filter, and use as the test solution.

[0219] The free sample solution and the total sample solution were collected and injected into the liquid chromatography system. The concentration of Plinabulin in the filtrate at different centrifugation times was calculated using the external standard method and the adsorption of the filter membrane was determined. The results are as follows:

[0220] Table 2-15 Filter membrane adsorption study - regenerated cellulose filter membrane

[0221] Through the adsorption study of dilution-resistant filter membrane, it can be seen that the regenerated cellulose filter membrane with a molecular weight cutoff of 30kd can reach adsorption saturation after 40 minutes.

[0222] To simulate clinical use, different dilution levels (1:10, 1:20, 1:30, and 1:50) were prepared and the diluted solutions were infused into 5L of pH 7.4 phosphate buffer over approximately 30 minutes (Note: The infusion time for the 1:50 dilution level solution is approximately 1 hour). The changes in encapsulation rate during the infusion process were studied. The results are as follows:

[0223] Table 2-16 Encapsulation rate results of solutions with different dilution levels

[0224] Based on the concentration of the total amount of the test solution during the infusion process, the actual infusion volume of the test sample and the concentration of 15-hydroxystearate polyethylene glycol ester at the corresponding time point were calculated. The results are as follows:

[0225] Table 2-17 Analysis of solution infusion results at different dilution levels

[0226] The results show that the encapsulation efficiency is low and varies significantly at the 1 / 3 pre-infusion volume. The critical micelle concentration (CMC) of 15-hydroxystearic acid polyethylene glycol ester (PEG) contained in the PEG excipient ranges from 0.005% to 0.02%. For reference, the CMC of PEG-15-hydroxystearic acid polyethylene glycol ester, measured by steady-state fluorescence, is 0.0035%. Combining the above infusion process and data, it can be seen that after 10 minutes of infusion of the (1:10) dilution level solution, only 17.6% of the actual infusion volume has been achieved. The PEG-15-hydroxystearic acid polyethylene glycol ester concentration in the PBS solution is at the CMC level, resulting in a low encapsulation efficiency. When the other dilution levels are 1 / 3 of the pre-infusion volume, the PEG-15-hydroxystearic acid polyethylene glycol ester concentration in the PBS solution is slightly higher than the CMC and the (1:10) dilution level, resulting in an encapsulation efficiency slightly above 50%-60%.

[0227] As the infusion time increased, the concentration of 15-hydroxystearate polyethylene glycol gradually increased, and the encapsulation efficiency also gradually improved. In 5L of pH 7.4 phosphate buffer, the final encapsulation efficiency of the test solution at different dilution levels was above 80%.

[0228] The encapsulation efficiency results show that when the solution at each dilution level is 1 / 3 of the pre-infusion concentration, the amount of 15-hydroxystearate polyethylene glycol in the PBS solution reaches or slightly exceeds the critical micelle concentration. The encapsulation efficiency is low. As the infusion time increases, the diluted solution gradually forms micelles in the buffer, and the encapsulation efficiency gradually increases to 80%.

[0229] Micellar stability study of diluted formulations

[0230] Batch samples of Plinabulin were diluted to dilution ratios of 1:10, 1:20, 1:30, and 1:50. The diluted solutions were stored at room temperature in the dark. Samples were taken at 0, 2, 4, 6, 8, 12, and 24 hours to study the stability of the micelles. The results are as follows:

[0231] Table 2-18 Encapsulation rate results at different dilution levels

[0232] The results showed that when the prepared test solutions of various dilution levels were stored in the dark at room temperature for 24 hours, the concentration of the total amount of the test solution within 0 hour was calculated to be 100.0%, and the concentration of the total amount of the test solution at each time point was in the range of 96.0% to 103.0%; the encapsulation rate was above 95%, and the micelle performance of the diluted solution was stable.

[0233] This product was diluted with D5W injection at different ratios (1:10, 1:20, 1:30, and 1:50), and the total amount of the test solution was stored at room temperature in the dark for 24 hours. There was no significant change in concentration and encapsulation rate, indicating that the micellar solution was stable at room temperature for 24 hours.

[0234] Determination of critical micelle concentration

[0235] The critical micelle concentration (CMC) is used to evaluate the micelle properties of a substance. Common measurement methods include surface tension method, conductivity method, fluorescent probe method, dye method, etc. Among them, the fluorescent probe method has been widely used in determining the critical micelle concentration of surfactants due to its advantages such as simple operation and little interference with the research system. Therefore, it was decided to use the fluorescent probe method to determine the concentration of 15-hydroxystearic acid polyethylene glycol ester in the formulation system. The critical microsphere concentration in 5% glucose injection and pH 7.4 PBS solution. The statistics of the equipment and reagents used in the study are as follows:

[0236] Pyrene mother liquor

[0237] Weigh 20 mg of pyrene as a fluorescent reagent into a 10 ml volumetric flask, dissolve it in acetone, dilute to the mark, and shake well. The concentration of pyrene in the solution is 2.0 mg / ml (1.0 × 10-5 mol / ml).

[0238] Propylene glycol-15-hydroxystearate polyethylene glycol mother liquor

[0239] Propylene glycol: 15-hydroxystearate polyethylene glycol = 60:40 (wt:wt). Weigh about 7.5g of propylene glycol and about 5g of 15-hydroxystearate polyethylene glycol, place them in a 50ml volumetric flask, add acetone to dissolve and dilute to the mark, shake well, and the concentration is 100mg / ml.

[0240] Sample preparation

[0241] Take 0.1 ml of pyrene mother liquor and place it in ten 10 ml volumetric flasks. After the acetone evaporates to dryness, add different amounts of propylene glycol-15-hydroxystearate polyethylene glycol mother liquor. After the acetone evaporates to dryness, dilute to the marked value with D5W injection solution and shake well. The concentration of 15-hydroxystearate polyethylene glycol micelles in the solution is 1×10 -7 g / ml, 5×10 -7 g / ml, 1×10 -6 g / ml, 5×10 -6 g / ml, 1×10 -5 g / ml, 5×10 -5 g / ml, 1×10 -4 g / ml, 5×10 -4 g / ml, 1×10 -3g / ml, 1×10 -2 g / ml. Measured after 24 hours at room temperature.

[0242] A series of solutions with the same concentration were prepared in parallel, diluted to the mark with PBS solution at pH 7.4, and placed at room temperature for 24 hours before measurement.

[0243] The specific measurement parameters of the fluorescence photometer are as follows:

[0244] Table 2-19 Fluorometer Setting Parameters

[0245] The fluorescence spectra of the two groups of solutions were measured according to the above method after being placed at room temperature for 24 hours.

[0246] The CMC values ​​of 15-hydroxystearate polyethylene glycol ester in different solution systems were obtained by curve fitting using the Boltzmann equation.

[0247] Table 2-20

[0248] The excipient specifications include a CMC range of 0.005% to 0.02%. The CMC of 15-hydroxystearate polyethylene glycol in water was determined to be 0.0035% as a control. There was no significant difference in the CMC of 15-hydroxystearate polyethylene glycol in different solution systems.

[0249] Example 3

[0250] This example describes the results of a dilution study conducted to determine the appearance, assay, impurity, and microbiological evaluation of Plinabulin (4 mg / mL) (propylene glycol / polyethylene glycol 15-hydroxystearate solution) diluted in 5% dextrose (D5W) in a non-PVC intravenous bag (500 mL) at various times after dilution. In this example, six (6) clear vials of 4 mg / mL Plinabulin samples and six (6) cloudy vials of 4 mg / mL Plinabulin samples were diluted with D5W to produce two dilution levels: approximately 1:20 and approximately 1:200.

[0251] A 4 mg / mL vial of Plinabulin was refrigerated and then diluted by heating at 37°C for 1 hour to reverse the turbidity. The diluted sample was initially tested at room temperature (0 hours) and then again after 4, 6, 8, 12, 24, and 48 hours of storage in the dark. The post-dilution stability and potential microbiological risk of Plinabulin (4 mg / mL) were determined after dilution with D5W in a non-PVC intravenous bag (500 ml) at 6, 8, 12, 24, and 48 hours.

[0252] Two lots of Plinabulin (4 mg / mL) were used in this study. The first lot was used for chemical and appearance analysis, and the second lot was used for microbial enumeration testing.

[0253] Filter interference studies were performed using the first batch of product at dilutions of 1:20 and 1:200. The 1:200 dilution did not meet the acceptance criteria for the filter interference studies, so unfiltered samples were used throughout the study.

[0254] For both the dilution studies and the microbial count studies, the results were identical for the clear vials and the vials with turbidity due to refrigeration. All samples met the acceptance criteria for microbial counts for both dilutions and all time points.

[0255] The 1:20 dilution met the assay stability criteria at 0, 4, 6, 8, and 12 hours. The 1:20 dilution failed to meet acceptance criteria at 24 and 48 hours post-dilution due to the presence of precipitate and a potency <90% of the label claim. The 1:200 dilution met the assay stability criteria at 0, 4, 6, and 8 hours post-dilution. The 1:200 dilution failed to meet acceptance criteria at 12, 24, and 48 hours post-dilution due to the presence of precipitate and a potency <90% of the label claim.

[0256] No impurities were detected at ≥ 0.10% in the clear and turbid vials at the 1:20 dilution level. For the 1:200 dilution level, the impurities present were below detection levels and were not quantified.

[0257] Filter Interference Studies

[0258] Possible interference from the 0.2 micron PBS filter was evaluated by filtering and analyzing a portion of the sample. A portion of the filtered sample was drawn from the port with the infusion tubing and compared to a portion of the unfiltered sample drawn directly from the outlet port. Three (3) samples were prepared and tested at each dilution level (1:20 and 1:200). A placebo control sample was prepared at the 1:20 dilution level to determine the placebo peak. For the 1:20 dilution level, the assay and impurity values ​​obtained from the filtered portion were compared to the values ​​from the unfiltered portion. For the 1:200 dilution, the assay values ​​obtained from the filtered portion were compared to the values ​​from the unfiltered portion. The results of the filtration interference study are listed in Table 3-1.

[0259] Dilution study method

[0260] In this study, 4 mg / mL Plinabulin Injection was diluted with D5W in a non-PVC IV bag to generate a diluted IV solution.

[0261] The volume of D5W solution contained in a single non-PVC IV bag (500 mL) was measured by pouring the bag into a 500 mL graduated cylinder or volumetric flask to the top, using an appropriately sized graduated cylinder with an accuracy of 1 mL or better. The average volume for sample calculations (n=4) was determined.

[0262] All D5W non-PVC N bags used for determination of mean volume were from the same manufacturer / supplier and from the same manufacturer's batch as all bags used for dilution studies.

[0263] The required volume of Plinabulin (4 mg / mL) is determined according to Formula 2. A nominal concentration of 0.2 mg / mL is obtained with a dilution factor of 1:20. For example, aseptically withdraw 12.5 mL (± 0.2 mL) from each of two (2) vials of Plinabulin (4 mg / mL) using a graduated syringe and inject directly into a 500 mL non-PVC IV bag covered with an amber sleeve. Record the weight of Plinabulin (4 mg / mL) injected into the bag. Determine the required volume of Plinabulin (4 mg / mL). A nominal concentration of 0.02 mg / mL is obtained with a dilution factor of 1:200. For example, aseptically withdraw 2.5 mL (± 0.05 mL) from a vial of Plinabulin (4 mg / mL) using a graduated syringe and inject directly into a 500 mL non-PVC IV bag covered with an amber sleeve. Record the weight of Plinabulin 4 mg / mL injected into the bag. Note: Impurities present were below detection levels and were not quantified at a 1:200 dilution.

[0264] Gently swirl the infusion bag to thoroughly mix the infusion. Inspect and record the appearance of the diluted solution in the non-PVC infusion bag. Store the diluted solution in the non-PVC infusion bag at controlled room temperature (25°C ± 3°C) in the dark for a total of 48 hours. Calculate the actual bag volume after injection according to Formula 3 and the label requirements after each dilution according to Formula 4. Inspect and record the appearance of the diluted solution at regular intervals. Samples are taken directly from the IV bag, not from the IV tubing. Chemical testing for assays and impurities is performed as described herein. Using the same IV tubing, samples are drawn directly through the tubing (without priming) and recorded at regular intervals. The first batch is used for chemical testing. Microbiological testing uses USP <61> The samples were drawn through the outlet port and recorded at each interval. The results of the dilution study are listed in Table 25. The second batch of product was used for microbial count studies.

[0265] Formula 2

[0266] Required volume of Plinabulin injection (4 mg / ml) = (C 静脉注射袋 ,mg / ml*V 静脉注射袋, ml) / (4mg / ml). Among them, C 静脉注射袋 is the desired final concentration of Plinabulin in the diluted IV bag, expressed in mg / ml (i.e., 0.2 mg / ml or 0.02 mg / ml), V 静脉注射袋 is the expected starting volume of D5W in the IV bag in ml, from the manufacturer's product instructions.

[0267] Formula 3

[0268] Actual volume 静脉注射袋 ,ml=V 平均 ,ml+((weight of Plinabulin injection (4mg / ml),g) / (d 普那布林注射液 (4mg / ml) ,g / ml)). Among them, V 平均 is the average volume measured from four IV bags, in ml. Wt. Plinabulin Injection (4 mg / ml) is the actual weight of Plinabulin Injection (4 mg / ml) added to the IV bag, in g. d Plinabulin Injection (4 mg / ml) is the density of Plinabulin Injection (4 mg / ml) obtained from the executed batch record.

[0269] Formula 4

[0270] Labeling requirements 静脉注射液 , mg / ml=(4mg / ml*Plinabulin injection (4mg / ml), g) / ((d intravenous injection (4mg / ml), g / ml*V 静脉注射袋 , ml). Where Wt Plinabulin Injection (4 mg / ml) refers to the actual weight added to the intravenous bag in g, d Plinabulin Injection (4 mg / ml) refers to the density of Plinabulin Injection (4 mg / ml) obtained from the batch execution record, and V 静脉注射袋 is the actual volume in the IV bag and is determined by Equation 2.

[0271] result

[0272] Filtering Interference Research

[0273] Results from the filter interference study at a 1:20 dilution level met the acceptance criteria of less than 3.0%. Results from the 1:200 dilution level did not meet the acceptance criteria of less than 3.0%. Because filter interference was observed at the 1:200 dilution level, unfiltered samples were used in the study. No impurities (~0.10%) were detected in either the filtered or unfiltered samples. The first batch of samples was used for testing.

[0274] Table 3-1 Filtering interference research

[0275] Dilution studies

[0276] The results of the dilution studies are presented in Tables 3-2, 3-3, and 3-4. Testing was performed using Batch 2. Microbiological testing was performed using Batch 1. Samples were drawn from the IV bag, tubing, or outlet port (as applicable).

[0277] Table 3-2 Dilution study

[0278] Table 3-3 Dilution study continued

[0279] Table 3-4 Dilution study continued

[0280] Table 3-5 Examples of microbial research

[0281] For the filter interference studies, a dilution of 1:200 did not meet the acceptance criteria, so unfiltered samples were used throughout the study.

[0282] For both the dilution studies and the microbial count studies, the results for the clear vials and the vials that became cloudy due to refrigeration were identical. All samples met the acceptance criteria for microbial counts for both dilutions and all time points.

[0283] The 1:20 dilution sample met the assay stability criteria at 0, 4, 6, 8, and 12 hours. The 1:20 dilution sample did not meet acceptance criteria at 24 and 48 hours after dilution due to the presence of precipitate and a potency < 90% of the label claim. The 1:200 dilution sample met the assay stability criteria at 0, 4, 6, and 8 hours after dilution. The 1:200 dilution sample did not meet acceptance criteria at 12, 24, and 48 hours after dilution due to the presence of precipitate and a potency < 90% of the label claim.

[0284] No impurities were detected at ≥0.10% in the clear and turbid vials at the 1:20 dilution level. For the 1:200 dilution level, the impurities present were below detection levels and were not quantified. Based on the results obtained from both the clear and turbid vials, it can be concluded that Plinabulin (4 mg / mL) diluted in D5W in a non-PVC IV bag (500 mL) is stable for 12 hours at a 1:20 dilution level and for 8 hours at a 1:200 dilution level when stored in the dark at room temperature.

[0285] All documents mentioned in this application are incorporated herein by reference, just as if each document were incorporated herein by reference individually. It should also be understood that after reading the above teachings of the present invention, those skilled in the art may make various changes or modifications to the present invention, and that such equivalents also fall within the scope of the claims appended hereto.

Claims

1. A method for preparing a Plinabulin micellar composition, characterized in that: The method comprises the steps of: s1) mixing polyethylene glycol 15-hydroxystearate and propylene glycol at 35° C.-65° C. to prepare a clear mixed solution; s2) mixing the clarified mixed solution in step 1) with Plinabulin at 35° C.-65° C. and stirring to prepare a Plinabulin micelle composition.

2. The preparation method according to claim 1, wherein The weight ratio of 15-hydroxystearate polyethylene glycol to propylene glycol is 1:5-5:1, preferably 1:3-3:1, more preferably 2:

3.

3. The preparation method according to claim 1, wherein In step (s2), Plinabulin is Plinabulin monohydrate.

4. A Plinabulin micellar composition, characterized in that The micelle composition comprises Plinabulin and a clear mixed solution of 15-hydroxystearate polyethylene glycol and propylene glycol, wherein the Plinabulin micelle composition is a yellow clear transparent solution, and the micelle particle size ranges from 10 to 100 nm.

5. The Plinabulin micellar composition according to claim 4, wherein The Plinabulin micelle composition is prepared by the method according to claim 1.

6. The Plinabulin micellar composition according to claim 4, wherein The Plinabulin micellar composition comprises the following ingredients: Propylene glycol 30-90wt%, preferably 40-70wt%, for example 50wt%, 55wt%, 60wt%, 65wt%, 70wt%; 20-60 wt % of polyethylene glycol 15-hydroxystearate, preferably 20-50 wt %, for example 25 wt %, 30 wt %, 35 wt %, 40 wt %, 45 wt %; and Plinabulin 1-10wt%, preferably 2-8wt%, such as 3wt%, 4wt%, 5wt%, 6wt%, 7wt%.

7. The Plinabulin micellar composition according to claim 4 or 6, wherein The weight ratio of 15-hydroxystearate polyethylene glycol to propylene glycol is 1:5-5:1, preferably 1:3-3:1, more preferably 2:

3.

8. A liquid Plinabulin composition for injection, comprising: Plinabulin, propylene glycol, and polyethylene glycol 15-hydroxystearate D5W solution (5% glucose injection); The volume ratio of propylene glycol to D5W in the composition is about 6:50 to about 6:

500.

9. The liquid Plinabulin composition for injection according to claim 8, characterized in that: The liquid Plinabulin composition for injection is prepared by the following method, which comprises the following steps: Providing an initial liquid formulation comprising plinabulin, propylene glycol and polyethylene glycol 15-hydroxystearate; and diluting the initial liquid formulation in D5W at a dilution ratio of about 1:5 to about 1:50; Wherein, the initial liquid preparation is the Plinabulin micelle composition as claimed in claim 4.

10. The liquid Plinabulin composition for injection according to claim 8, characterized in that: The concentration of Plinabulin is about 0.08 mg / ml to about 0.4 mg / ml.

11. The liquid Plinabulin composition for injection according to any one of claims 8 to 10, characterized in that The volume ratio of 15-hydroxystearate polyethylene glycol ester and D5W in the liquid injection Plinabulin preparation is about 4:50 to about 4:500, preferably about 4:100 to about 4:500, more preferably about 4:100 to about 4:400, 12. The liquid Plinabulin composition for injection according to claim 8, characterized in that: The total amount of impurities contained in the liquid Plinabulin composition for injection is less than 0.5%.

13. The liquid Plinabulin composition for injection according to claim 8, wherein Greater than about 90% of the Plinabulin in the composition is encapsulated in the micelles.

14. A method for preparing a liquid Plinabulin composition for injection, characterized in that: The method comprises: Providing an initial liquid formulation comprising plinabulin, propylene glycol and polyethylene glycol 15-hydroxystearate; and The initial liquid formulation was diluted in D5W at a dilution ratio of about 1:5 to about 1:

50.

15. The method for preparing a liquid Plinabulin composition for injection according to claim 14, wherein: The initial liquid preparation is the Plinabulin micellar composition according to claim 4.

16. Use of the Plinabulin micelle composition according to claim 4 or the Plinabulin composition for liquid injection according to claim 8 in the preparation of an anti-tumor drug.

17. The use according to claim 16, characterized in that The tumor is selected from the group consisting of lung cancer (such as small cell lung cancer, non-small cell lung cancer), prostate cancer, colon cancer, brain tumor (such as glioblastoma, glioblastoma multiforme, giant cell glioblastoma, metastatic brain tumor), head and neck cancer, gastric cancer, pancreatic cancer, breast cancer, kidney cancer, bladder cancer, ovarian cancer, cervical cancer, melanoma, myeloma, lymphoma or leukemia.

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