CN115869314A

CN115869314A - Composition containing plinabulin and preparation method thereof

Info

Publication number: CN115869314A
Application number: CN202211187926.7A
Authority: CN
Inventors: 严新宇; 邹爱峰; 仝新勇
Original assignee: Jiangsu Hengrui Medicine Co Ltd; Shanghai Hengrui Pharmaceutical Co Ltd
Current assignee: Jiangsu Hengrui Medicine Co Ltd; Shanghai Hengrui Pharmaceutical Co Ltd
Priority date: 2021-09-29
Filing date: 2022-09-28
Publication date: 2023-03-31

Abstract

The present disclosure relates to plinabulin-containing compositions and methods of making the same. In particular, the disclosure relates to compositions comprising plinabulin and a phospholipid and methods of making the same to provide a therapeutically effective dose of plinabulin to improve leukopenia compliance.

Description

Composition containing plinabulin and preparation method thereof

Technical Field

The disclosure belongs to the field of pharmacy, and relates to a composition containing plinabulin and a preparation method thereof.

Background

Plinabulin is a synthetic analog of the diketopiperazine phenyl aspartame (benzytrimethylammonium chloride) found in marine and terrestrial aspergillus species. 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 the tubulin monomer. Previous studies showed that plinabulin at low concentrations, compared to colchicine, induced tubulin depolymerization and monolayer permeability of vascular endothelial cells, and that plinabulin induced apoptosis in Jurkat leukemia cells. The study of plinabulin as a single agent in patients with advanced malignancies (lung, prostate and colon cancers) showed good pharmacokinetic, pharmacodynamic and safety profiles.

CN105705148 discloses a plinabulin preparation, which further comprises 40% polyethylene glycol-15 hydroxystearate HS-15 and 60% propylene glycol, and the drug concentration is 4mg/mL. However, it is understood that HS-15 is not an absolutely safe adjuvant. Ever marketed product Lapidan fat emulsion

The medium HS-15 dosage is 4.4%, and the single dosage is 4.07g according to a single dose of 92.5 mL/bottle, and is close to the preparation disclosed in CN 105705148. But which>

Fat milk has been removed from the market due to severe allergic reactions that occur after it is marketed.

Disclosure of Invention

The present disclosure relates to a composition comprising plinabulin or a pharmaceutically acceptable salt thereof and at least one lipid.

In some embodiments, the lipid comprises at least one phospholipid.

The present disclosure relates to a medicament for intravenous injection, which is a plinabulin composition, comprising plinabulin and a lipid, the lipid comprising at least one phospholipid.

The lipids of the present disclosure may also comprise other neutral, cationic and/or anionic lipids.

Examples of other neutral lipids that can be used in the present disclosure include: steroid such as cholesterol and its derivatives, cephalin, sphingomyelin, and hydrogenated soybean phospholipid.

The phospholipid is selected from the group consisting of phosphatidylcholine, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol, phosphatidylserine, phosphatidic acid, lysophospholipid, sphingomyelin, lecithin and cardiolipin in any form, including salted or desalted, hydrogenated or partially hydrogenated, natural, semi-synthetic or synthetic forms. The lipids described in the present disclosure include completely neutral or negatively charged phospholipids. The term "phospholipid" refers to a hydrophobic molecule containing at least one phosphorus group, which may be natural or synthetic. For example, the phospholipid may comprise a phosphorus-containing group and a saturated or unsaturated alkyl group optionally substituted with OH, COOH, oxo, amine, or a substituted or unsubstituted aryl group. Phospholipids differ from each other in the length of their acyclic (acrylic) chains and in the degree of unsaturation. The term "phosphatidylcholine" refers to phosphatidylcholine and derivatives thereof.

In some embodiments of the present invention, the substrate is, examples of phospholipids suitable for use in the present disclosure include any form of Dipalmitoylphosphatidylcholine (DPPC), distearoylphosphatidylcholine (DSPC), dimyristoylphosphatidylcholine (DMPC), 1-palmitoyl-2-lauroyl-sn-glycero-3-phosphatidylcholine (PLPC), phosphorylcholine (DOPC), egg yolk lecithin (EPC), dilauroylphosphatidylcholine (DLPC), hydrogenated Soy Phosphatidylcholine (HSPC), l-myristoyl-2-palmitoyl phosphatidylcholine (MPPC), l-palmitoyl-2-myristoylphosphatidylcholine (PMPC), l-palmitoyl-2-stearoylphosphatidylcholine (PSPC) l-stearoyl-2-palmitoyl phosphatidylcholine (SPPC), palmitoyl Oleoyl Phosphatidylcholine (POPC), lysophosphatidylcholine, dilinoleoyl phosphatidylcholine, distearoyl phosphatidylethanolamine (DSPE), distearoyl phosphatidylethanolamine-polyethylene glycol 2000 (MPEG 2000-DSPE), dimyristoyl phosphatidylethanolamine (DMPE), dipalmitoyl phosphatidylethanolamine (DPPE), dioleoyl phosphatidylglycerol (DOPG), dimyristoyl phosphatidylglycerol (DMPG), distearoyl phosphatidylglycerol (DSPG), dipalmitoyl glycerophosphoglycerol (DPPG), dipalmitoyl phosphatidylserine (DPPS), 1, 2-dioleoyl-sn-glycero-3-phosphatidylserine (DOPS), dimyristoylphosphatidylserine (DMPS), distearoylphosphatidylserine (DSPS), dipalmitoylphosphatidic acid (DPPA), 1, 2-dioleoyl-sn-glycero-3-phosphatidic acid (DOPA), dimyristoylphosphatidic acid (DMPA), distearoylphosphatidic acid (DSPA), dipalmitoylphosphatidylglycerol (DPPI), 1, 2-dioleoyl-sn-glycero-3-phosphatidylinositol (DOPI), dimyristoylphosphatidylglycerol (DMPI), distearoylphosphatidylinositol (DSPI), (2, 3-dioleoyl-propyl) -trimethylamine (DOTAP), soybean lecithin, any of which forms includes one or more of salted or desalted, hydrogenated or partially hydrogenated, natural, semisynthetic or synthetic forms.

In some embodiments, the phospholipids in the present disclosure are selected from at least one of DOPC and egg yolk lecithin.

In certain embodiments, the lipid comprises at least one phospholipid and cholesterol.

In certain embodiments, the lipid comprises at least one phosphatidylcholine and cholesterol.

In some embodiments, the composition comprises plinabulin, cholesterol, and at least one phosphatidylcholine.

In some embodiments, the liposome composition comprises plinabulin, cholesterol, and DOPC.

In some embodiments, the composition comprises plinabulin and a phospholipid. The weight ratio of plinabulin to phospholipid is selected from 1. In some embodiments, the ratio of 1.

In some embodiments, the composition further comprises a steroid. In some embodiments, the steroid is selected from cholesterol.

In some embodiments, the weight ratio of cholesterol to phospholipid is selected from 1.

In some embodiments, the weight ratio of cholesterol to phospholipid is selected from 1 to 20.

In some embodiments, the composition as previously described, comprises a weight ratio of plinabulin to phospholipid selected from 1 to 30-100, and a weight ratio of cholesterol to phospholipid selected from 1 to 5-20.

In some embodiments, the compositions of the present disclosure further comprise water or an aqueous solution. In certain embodiments, the aqueous solution includes, but is not limited to, an isotonic solution or buffer. The isotonic solution has a solution with an osmotic pressure substantially the same as human blood, and can be prepared by an isotonic agent and water. The isotonic agent comprises sodium chloride, potassium chloride, magnesium chloride, calcium chloride, glucose, xylitol and sorbitol. The buffer refers to a buffered solution that resists changes in pH by the action of its acid-base conjugate components. In one embodiment, the pH of the buffer of the present disclosure ranges from about 4.5 to about 8.5. Examples of buffers that can control the pH in this range include acetate (e.g., sodium acetate), succinate (e.g., sodium succinate), gluconate, histidine, citrate, carbonate, PBS, HEPES, or other organic acid buffers.

In the composition disclosed by the disclosure, the particle size of 200nm polycarbonate membrane filtration is less than 500nm. In some embodiments, the particle size is less than 400nm. In some embodiments, the particle size is less than 300nm. In some embodiments, the particle size is less than 250nm. In some embodiments, the particle size is less than 200nm.

In some embodiments, the plinabulin composition is in unit dosage form, comprising plinabulin 1-60mg. In some embodiments, the plinabulin composition is in unit dosage form, comprising 10-50mg of plinabulin. In some embodiments, the plinabulin composition is in unit dosage form, comprising plinabulin 10mg, 11mg, 12mg, 13mg, 14mg, 15mg, 16mg, 17mg, 18mg, 19mg, 20mg, 21mg, 22mg, 23mg, 24mg, 25mg, 26mg, 27mg, 28mg, 29mg, 30mg, 31mg, 32mg, 33mg, 34mg, 35mg, 36mg, 37mg, 38mg, 39mg, 40mg, 41mg, 42mg, 43mg, 44mg, 45mg, 46mg, 47mg, 48mg, 49mg, 50mg. In some embodiments, the plinabulin composition is in unit dosage form, comprising 20mg, 40mg of plinabulin.

In some embodiments, the concentration of plinabulin in the composition is selected from 0.1-10mg/mL, and can be 0.1mg/mL, 0.2mg/mL, 0.3mg/mL, 0.4mg/mL, 0.5mg/mL, 0.6mg/mL, 0.7mg/mL, 0.8mg/mL, 0.9mg/mL, 1mg/mL, 1.1mg/mL, 1.2mg/mL, 1.3mg/mL, 1.4mg/mL, 1.5mg/mL, 1.6mg/mL, 1.7mg/mL, 1.8mg/mL, 1.9mg/mL, 2.0mg/mL.

In some embodiments, the concentration of plinabulin in the composition is selected from 0.1-2mg/mL.

In some embodiments, the concentration of plinabulin in the composition is selected from 0.1-1mg/mL.

The disclosure also provides the use of the plinabulin composition or the plinabulin composition prepared by the method in the preparation of a medicament for preventing or treating diseases and conditions in which an individual suffers from neutropenia.

The present disclosure provides a method of preventing or treating diseases and disorders in an individual suffering from neutropenia comprising administering to the subject a therapeutically effective amount of a plinabulin composition.

The present disclosure provides a method of preventing or treating diseases and disorders of neutropenia in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a plinabulin composition.

In certain embodiments, the plinabulin composition is administered in a dosage range selected from 1-400mg, 1-300mg, 10-200mg, 10-160mg, 10-140mg, 10-120mg, 10-100mg, 10-80mg, 10-60mg, or 10-40mg.

In some embodiments, the plinabulin composition is administered in a daily dose range selected from 1 to 100mg. In some embodiments, the plinabulin composition is administered in a daily dose range selected from the group consisting of 1-80mg, 1-60mg, 1-40mg, 10-30mg, 10-20mg.

The plinabulin composition of the present disclosure can be prepared by a thin film evaporation method, a freeze-drying method, an active drug loading method, a pH gradient method, an injection method, and/or an ultrasonic dispersion method. In some embodiments, a step of size stabilization (or homogenization, homogenization) may be further included, and the size stabilization method may be, but is not limited to, using a high pressure homogenizer, sonication, extrusion, and the like.

In some embodiments, the plinabulin composition is prepared by an infusion method, specifically comprising: dissolving phospholipid and steroid and plinabulin in organic solvent, mixing with water or water solution, removing organic solvent, and grading.

In certain embodiments, the aqueous solution includes, but is not limited to, an isotonic solution or buffer.

In some embodiments, the plinabulin composition is prepared by a thin film evaporation method, specifically including: dissolving plinabulin, phospholipid and steroid in organic solvent, removing solvent, forming film, and hydrating, wherein in some embodiments, the preparation method further comprises extruding and granulating the liposome composition obtained after hydration.

The plinabulin compositions of the present disclosure can be in the form of any form of a mixture of plinabulin with lipids, including in the form of liposomes.

The plinabulin composition of the present disclosure may be further dried or otherwise removed from the liquid medium in the system to obtain a dry solid composition. Methods of drying the composition include, but are not limited to, evaporation, freeze drying, spray drying, or the like.

The plinabulin compositions of the present disclosure can be administered in a form selected from the group consisting of intravenous injection, subcutaneous injection, and tissue injection, by parenteral injection. Administration of the liposome compositions of the present disclosure is accomplished using standard methods and devices, e.g., pens, syringe systems, needles and syringes, hypodermic delivery systems, catheters, and the like.

The term "about" means that amounts, sizes, formulations, parameters, and other quantities and characteristics are not and need not be exact, but may be approximate and/or larger or smaller as desired, reflecting tolerances, conversion factors, rounding off, measurement error and the like, and other factors known to those of skill in the art. The meaning may include a variation of. + -. 10%, preferably. + -. 5%.

Detailed Description

The present disclosure will be explained in more detail with reference to examples, which are only used for illustrating the technical solutions of the present disclosure and do not limit the spirit and scope of the present disclosure.

The particle size disclosed by the disclosure adopts a Malvern Nano particle size analyzer, model No-ZS, and the average particle size is Intensity Mean Diameter. The assay described in this disclosure uses a high performance liquid chromatograph, model number Thermo U300 (VWD + CAD), sourced from Thermo Fisher Scientific, usa. And a one-tenth-ten-thousandth electronic balance is adopted for weighing the reference substance, the model is 225D-1CN, and the source is Sidoristes. The column type of the column used for the assay was Waters Xbridge C ₁₈ 5 μm, 4.6X 150mm, from Waters corporation, USA. The diluent was Merk, a methanol source. Triethylamine used for preparing the mobile phase is CNW (CNW) which is an analytically pure source, phosphoric acid is a Chinese medicine reagent which is an analytically pure source, purified water is prepared by self, and acetonitrile is obtained from Merk.

Example 1: plinabulin composition under formula 1 DOPC system

Components	Dosage of prescription
		Plinabulin	40mg
DOPC	3.144g
		Cholesterol	172mg
Ethanol	10mL
		5% glucose injection	200mL

Prepared by ethanol injection method: the prescribed amount of plinabulin, DOPC and cholesterol were taken and placed in a beaker, stirred and dissolved in absolute ethanol (adjuvant grade, nanjing chemical reagents ltd). Slowly dripping the drug-lipid solution into 5% glucose injection (prepared by laboratory) at 40 deg.C, and stirring thoroughly. The solution was transferred to a round bottom flask and ethanol was removed by rotary evaporation. Then, the drug solution was passed through a 0.2 μm polycarbonate membrane (ATS Nano reagent machine, model: AH 100D) at 50 ℃ to form a uniform and transparent liposome solution.

Test example 1: index associated with prescription 1

Through observation, the polycarbonate membrane almost has no plinabulin API, and the sampling is detected by a nanometer particle size analyzer, so that the particle size of the liquid medicine is 56.90nm; the drug loading was 89.8%.

The detailed measurement procedure is as follows:

preparing a reference solution: taking and precisely weighing about 25.0mg of plinabulin reference substance, placing the reference substance in a 25ml volumetric flask, firstly adding a proper amount of methanol for ultrasonic dissolution, cooling to room temperature, adding methanol for dilution to a constant volume to scale, and shaking up to obtain the reference substance stock solution. Precisely transferring 1ml of the reference substance stock solution into a 25ml volumetric flask, adding methanol for dilution and fixing the volume to the scale to obtain the reference substance solution of the Plinabulin.

Preparing a content determination test solution: measuring 1ml of the solution by using a liquid transfer gun, placing the solution in a 10ml volumetric flask, adding methanol to dissolve and fix the volume to a scale, and shaking up to obtain the product.

Content determination chromatographic conditions: the mobile phase is triethylamine phosphate buffer solution-acetonitrile gradient elution, wherein the preparation method of the triethylamine phosphate buffer solution comprises the steps of measuring 7.5ml of triethylamine, adding water to dilute the triethylamine phosphate buffer solution to 1000ml, and adjusting the pH value to 3.0 +/-0.5 by using phosphoric acid. The gradient elution procedure is shown in the following table, with a detection wavelength of 240nm, a column temperature of 30 deg.C, a flow rate of 1.0ml/min, a sample size of 10. Mu.L, and a run time of 15min.

Time/min	Triethylamine phosphate buffer solution	Acetonitrile (ACN)
			0	75	25
6	60	40
			11	20	80
13	75	25
			15	75	25

Example 2: prescription 2 Plinabulin composition under yolk lecithin system

Components	Dosage of prescription
		Plinabulin	40mg
Yolk lecithin PL-100M	3.6g
		CHOL	0.54g
API	40mg
		5% glucose	100mL
Methylene dichloride	50mL

Prepared by thin film evaporation: prescription amounts of plinabulin, egg yolk lecithin, and cholesterol were placed in a round bottom flask and dissolved with stirring in 99.9% dichloromethane (HPLC grade, damas-beta). The solution was rotary evaporated to remove methylene chloride and form a film. The films were hydrated by pouring a 5% glucose solution into a round bottom flask at 50 ℃. Then, the drug solution was passed through a 0.2 μm polycarbonate membrane to form a uniform and transparent liposome solution.

Test example 2: prescription 2 related index

Through observation, a layer of plinabulin API is arranged on the polycarbonate film, and the granularity of the liquid medicine is 37.55nm and the medicine-loading rate is 16.7 percent by detecting with a nanometer particle size analyzer after sampling (the determination method is the same as that of test example 1).

Comparative example 1: investigation result of micellar system on solubilization of plinabulin

Experimental results show that the solubilization effect of the micelle system is poor.

Comparative example 2: investigation result of solubilization effect of cosolvent on plinabulin

1) Solubilizing effect of meglumine:

154mg of meglumine was dissolved in 10mL of a 5% glucose solution, 20mg of plinabulin was added, and the mixture was stirred at 50 ℃ for 30min, whereby the plinabulin was completely insoluble.

2) Solubilization effect of hydroxypropyl-beta-cyclodextrin:

1.26g of hydroxypropyl-beta-cyclodextrin is dissolved in 10mL of purified water, 40mg of plinabulin is added into the purified water at the temperature of 50 ℃, the mixture is fully stirred for 30min, the solution is slightly yellow, a large amount of plinabulin solid exists, and the solubilizing effect is poor.

Comparative example 3: investigation result of solubilization of Plinabulin by microemulsion

Detection shows that 10mg of plinabulin is not dissolved in 5mL of refined castor oil, medium-chain triglyceride, and isopropyl myristate.

Claims

1. A composition comprising plinabulin or a pharmaceutically acceptable salt thereof and at least one lipid.

2. The composition of claim 1, wherein the lipid comprises at least one phospholipid.

3. The composition of claim 2, wherein the phospholipid is selected from the group consisting of phosphatidylcholine, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol, phosphatidylserine, phosphatidic acid, lysophospholipid, sphingomyelin, lecithin, and cardiolipin.

4. The composition of matter as claimed in claim 2, the phospholipid is selected from Dipalmitoylphosphatidylcholine (DPPC), distearoylphosphatidylcholine (DSPC), dimyristoylphosphatidylcholine (DMPC), 1-palmitoyl-2-lauroyl-sn-glycero-3-phosphatidylcholine (PLPC), phosphatidylcholine (DOPC), egg yolk lecithin (EPC), dilauroylphosphatidylcholine (DLPC), hydrogenated Soybean Phosphatidylcholine (HSPC), l-myristoyl-2-palmitoyl phosphatidylcholine (MPPC), l-palmitoyl-2-myristoylphosphatidylcholine (PMPC), l-palmitoyl-2-stearoylphosphatidylcholine (PSPC), l-stearoyl-2-palmitoylphosphatidylcholine (SPPC), palmitoyl Oleoylphosphatidylcholine (POPC), lysophosphatidylcholine, dilinoleoylphosphatidylethanolamine (DSPE), distearoylphosphatidylethanolamine-polyethylene glycol 2000 (MPEG 2000-DSPE), dimyristoylphosphatidylethanolamine (DMPE), dipalmitoylphosphatidylethanolamine (DPPE), dioleoylphosphatidylglycerol (DPG), dimyristoylphosphatidylglycerol (DMPG), distearoylphosphatidylglycerol (DOPPG), distearoylphosphatidylglycerol (DPPS), dimyristoylphosphatidylcholine (DPPG), and DPPG, 1, 2-dioleoyl-sn-glycero-3-phosphatidylserine (DOPS), dimyristoylphosphatidylserine (DMPS), distearoylphosphatidylserine (DSPS), dipalmitoylphosphatidic acid (DPPA), 1, 2-dioleoyl-sn-glycero-3-phosphatidic acid (DOPA), dimyristoylphosphatidic acid (DMPA), distearoylphosphatidic acid (DSPA), dipalmitoylphosphatidylglycol (DPPI), 1, 2-dioleoyl-sn-glycero-3-phosphatidylinositol (DOPI), dimyristoylphosphatidylglycol (DMPI), distearoylphosphatidylinositol (DSPI), (2, 3-dioleoyl-propyl) -trimethylamine (DOTAP), soybean lecithin, preferably at least one of DOPC and egg yolk.

5. The composition according to claim 2, wherein the weight ratio of plinabulin to phospholipid is selected from 1.

6. The composition according to claim 2, the lipid further comprising at least one steroid, preferably cholesterol.

7. The composition of claim 6, wherein the weight ratio of cholesterol to phospholipid is selected from 1; preferably 1.

8. The composition of any of claims 1-7, comprising a weight ratio of plinabulin to phospholipid selected from 1.

9. The composition according to any of claims 1 to 8, in unit dosage form, comprising 1 to 60mg of plinabulin, preferably 10 to 50mg, more preferably 20 to 40mg.

10. The composition according to any of claims 1-9, wherein the concentration of plinabulin is selected from 0.1-10mg/mL, preferably 0.1-2mg/mL, more preferably 0.1-1mg/mL.

11. Composition according to any of claims 1 to 10, having an average particle size of less than 500nm, preferably less than 300nm.

12. A composition according to any of claims 1-11, said composition comprising plinabulin, cholesterol, and at least one phosphatidylcholine, preferably comprising plinabulin, cholesterol, and DOPC.

13. A method of preparing the composition of claims 1-12, comprising thin film evaporation, freeze drying, active drug loading, pH gradient, injection, or ultrasonic dispersion.

14. The method according to claim 13, which is an injection method, comprising: dissolving phospholipid and steroid with Plinabulin in organic solvent, mixing with water or water solution, removing organic solvent, and grading.

15. The method according to claim 13, which is a thin film evaporation method, comprising: dissolving plinabulin, phospholipid and steroid in organic solvent, removing solvent, forming film, and hydrating.

16. A composition according to any one of claims 1 to 12 or prepared by a method according to any one of claims 13 to 15 in the form of liposomes.

17. Use of the composition of any one of claims 1-12 or the composition prepared by the method of any one of claims 13-15 or the liposome of claim 16 in the manufacture of a medicament for preventing or treating diseases and conditions in which a subject suffers from neutropenia.