CN106852911B

CN106852911B - Stable cabazitaxel albumin composition and preparation method thereof

Info

Publication number: CN106852911B
Application number: CN201611107855.XA
Authority: CN
Inventors: 苏正兴; 杨立开; 赵栋; 周晶; 王利春; 王晶翼
Original assignee: Sichuan Kelun Pharmaceutical Research Institute Co Ltd
Current assignee: Sichuan Kelun Pharmaceutical Research Institute Co Ltd
Priority date: 2015-12-07
Filing date: 2016-12-06
Publication date: 2020-03-17
Anticipated expiration: 2036-12-06
Also published as: CN106852911A

Abstract

The invention provides a stable cabazitaxel sterile freeze-drying composition and a preparation method thereof. Through screening of a preparation formula and a preparation process, the cabazitaxel albumin composition prepared by using the low-concentration albumin solution has the characteristics of excellent stability and safety, a water-insoluble chloroform/dichloromethane and other high-toxicity organic solvents are not required to be used in the preparation process, a freeze-drying protective agent and a protein stabilizing agent are not required to be added in the freeze-drying step, the production cost of the medicine is reduced, and the safety of clinical medication is ensured.

Description

Stable cabazitaxel albumin composition and preparation method thereof

Technical Field

The present invention relates to the field of pharmaceutical formulations. More particularly, the invention relates to a stable cabazitaxel albumin sterile freeze-dried composition and a preparation method thereof.

Background

Prostate cancer is the most common malignant tumor of the male reproductive system, the incidence rate of the prostate cancer increases with age, the incidence rate of the prostate cancer has obvious regional difference, the incidence rate is higher in European and American areas, and the incidence rate is lower in Asian areas. The incidence and mortality of prostate cancer is second only to lung cancer, and is the second leading cause of cancer death. In most cases, prostate cancer develops slowly in older men, and in asia, its incidence is lower than in western countries, but in recent years there is a rapidly rising trend.

The american cancer society has counted that prostate cancer accounts for one-fourth of the newly diagnosed cases of cancer annually in the united states, about 22 million patients with prostate cancer diagnosed in 2010, and over 32,000 patients die of the disease. In europe, the annual number of deaths is 27,000, but worldwide the number of prostate cancer deaths far exceeds this number. Since the onset of prostate cancer is often late, the patient population has a tendency to age. The prostate cancer medicament has wide market prospect. The market research company Decision Resources projected that the market value of prostate cancer drugs would increase from nearly $ 40 million in 2009 to $ 57 million in 2014; the year 2019 will further increase to $ 87 billion.

Currently, there are several common medications for prostate cancer:

(1) anti-androgen therapy: antiandrogen preparations act primarily on the blockade of the receptors of prostate cells by androgens, so that cancer cells cannot receive androgen messages and do not continue to grow, and antiandrogen drugs include: flutamide tablets, compacter (bicalutamide tablets), and the like.

(2) Chemotherapy: the traditional Chinese medicine composition can be used as adjuvant therapy and often has the effect that endocrine therapy and radiotherapy cannot replace the effect. The most sensitive chemotherapy medicine for prostate cancer is compound cyclophosphamide tablet, and other chemotherapy medicines include cisplatin (PDD), 5-fluorouracil (5-FU), Adriamycin (ADM), mitomycin (MMC), estramustine phosphate (EMP), taxanes, etc.

(3) Treatment with traditional Chinese medicines: has certain treatment effect according to different pulse phases for syndrome differentiation treatment.

(4) The immune therapy is that the relation between immunity and tumor is very interesting in recent years, the immune drug inhibits the growth of tumor cells by improving the immunity of the organism and disease resistance, and simultaneously, the immune drug can also protect the cell function and reduce the damage of endocrine drugs and anticancer drugs to the organism, and Keyin probiotics (recombinant human interferon α 2b injection), Xingill (recombinant human interleukin-2 (125Ala) for injection) and the like are clinically applied at present.

(5) Bisphosphonate drug therapy: the method is mainly applied to prostate cancer bone metastasis patients for targeted therapy and the like.

Cabazitaxel (cabazitaxel) is an antitumor drug, belongs to taxanes, and is prepared by semi-synthesizing precursors extracted from taxus chinensis.

Cabazitaxel has a molecular formula of C₄₅H₅₇NO₁₄White crystals, insoluble in water and soluble in ethanol, with the chemical name (2R,3S) -3-tert-butoxycarbonylamino-2-hydroxy-3-phenylpropionic acid-4 α -acetyl-2 α -benzoyl-5 β, 20-epoxy-1 β -hydroxy-7 β,10 β -dimethoxy-9-oxo-11-taxen-13 α -ester, the structural formula of which is shown below:

on 17 days 6 months 2010, cabazitaxel was approved by the U.S. Food and Drug Administration (FDA) as a second line therapy for the treatment of prostate cancer. Cabazitaxel belongs to an anti-microtubule drug on an anti-tumor mechanism, promotes tubulin to polymerize into microtubules mainly by combining with free tubulin, simultaneously prevents assembled microtubules from disintegrating to form stable non-functional microtubules, and further inhibits the mitosis and the tumor cell function at the interphase of cells. Cabazitaxel has a very good curative effect on prostate cancer, and can be used together with prednisone to treat prostate cancer patients who are refractory to metastatic hormone in a previous docetaxel-containing treatment scheme.

In terms of physicochemical properties, cabazitaxel has very poor water solubility, about 6 μ g/ml, as other taxanes. When the cabazitaxel injection is clinically used, the cabazitaxel injection is dissolved in tween-80 and diluted by ethanol, glucose or normal saline for injection, which not only brings inconvenience to clinical application, but also causes a series of safety problems due to the introduction of auxiliary materials such as tween-80 and the like.

Disclosure of Invention

In one aspect, the invention provides a safe and stable albumin drug sterile lyophilized composition of cabazitaxel.

The composition of the present invention is a nanoparticle system consisting of cabazitaxel or derivatives thereof, such as solvates, and albumin.

In the pharmaceutical composition, the mass ratio of cabazitaxel or derivatives thereof to albumin is 1: 2-30, the pH is 6.0-7.5, the average particle size is not more than 200nm, and the zeta potential of the freeze-dried composition is-2 mv to-9 mv, wherein a freeze-drying protective agent and a protein stabilizing agent are not required to be added in the freeze-drying process.

Preferably, the mass ratio of the cabazitaxel or the derivatives thereof to the albumin is 1: 10-20, and more preferably 1: 12-17;

preferably, in the sterile lyophilized pharmaceutical composition consisting of cabazitaxel or derivatives thereof and albumin, the pH is 6.5-7.0;

in the pharmaceutical composition of cabazitaxel or derivatives thereof and albumin, the zeta potential of the composition after freeze-drying is between-5 mv and-8 mv, wherein the re-dissolving solvent comprises physiological saline, for example.

Preferably, the albumin is a protein with a sulfhydryl group or a disulfide bond, preferably human serum albumin.

In another aspect, the present invention provides a method for preparing the sterile lyophilized composition of cabazitaxel and albumin, comprising the following steps:

1) dissolving cabazitaxel or derivatives thereof, such as solvates, in an organic solvent to form an organic phase; the organic solvent is selected from alcohol solvents, and the concentration of cabazitaxel or derivatives thereof in the organic phase is 10-45 mg/ml;

2) dispersing the albumin in an aqueous medium to form an aqueous phase; in the water phase, the concentration of albumin is 3-20 mg/ml;

3) mixing the organic phase obtained in the step 1) and the water phase obtained in the step 2), shearing at a high speed, and homogenizing to obtain a suspension with the average particle size of not more than 200 nm; wherein, the homogenization can adopt a mode of transferring the mixture into a high-pressure homogenizer, for example;

4) carrying out ultrafiltration concentration on the suspension obtained in the step 3) to obtain an albumin composition of cabazitaxel;

5) filtering and sterilizing the composition obtained in the step 4), and then directly freezing and drying to obtain the sterile freeze-dried composition of cabazitaxel and albumin.

In the preparation method, the freezing process in the step 5) does not need to add a lyoprotectant and a protein stabilizer, wherein the lyoprotectant comprises, but is not limited to, trehalose, sucrose, maltose, lactose and mannitol; wherein the protein stabilizer includes but is not limited to one or more of trehalose, sucrose, sodium caprylate, amino acids (e.g., acetyl tryptophan, glycine, etc.).

Preferably, the alcohol solvent in step 1) is selected from methanol, ethanol, ethylene glycol, isopropanol, and more preferably ethanol;

preferably, the concentration of albumin in the step 2) is 3-15 mg/ml;

preferably, ultrafiltration concentration in the step 4) is 2-20 times;

preferably, the concentration of the cabazitaxel or the derivatives thereof in the steps 3) and 4) is 0.1-6 mg/ml; preferably 0.4-4 mg/ml;

preferably, the shearing is carried out at the rotating speed of 7000-15000 rpm in the step 3);

preferably, the high-pressure homogenization is carried out under the pressure of 30000-40000 psi in the step 3);

preferably, the homogenization step in the step 3) is repeated for 5 to 6 cycles.

On the other hand, the invention also provides the sterile freeze-drying composition of the cabazitaxel and the albumin, which is prepared by the preparation method.

The cabazitaxel albumin freeze-dried composition prepared by the preparation method provided by the invention has good stability, for example, the stability of the composition is evaluated by measuring the content change or the particle size change or the zeta potential change of the cabazitaxel in a certain time period, and the stability of the freeze-dried composition prepared by the preparation method provided by the invention is proved to be more than 72 hours in the aspects.

In another aspect, the present invention further provides a formulation comprising the above-mentioned sterile lyophilized composition of cabazitaxel and albumin, and a pharmaceutically acceptable carrier and/or adjuvant.

As a preferred aspect of the present invention, the pharmaceutically acceptable carriers and/or excipients used in the present invention include sterile water, saline, dextrose, oils (e.g., corn oil; peanut oil, sesame oil, and the like), acids, lower alkanols (glycols; polyalkylene glycols).

As a preferable aspect of the present invention, the pharmaceutically acceptable carrier and/or adjuvant used in the present invention may further include preservatives, wetting agents, emulsifiers, penetration enhancers, and the like.

Preferably, the formulations of the present invention may further comprise an antimicrobial agent (e.g., a chelating agent, including but not limited to EDTA, edetate, citrate, pentetate, tromethamine, sorbate, ascorbate, derivatives thereof or mixtures thereof; e.g., a non-chelating agent, including but not limited to any one or more of sulfite, benzoic acid, benzyl alcohol, chlorobutanol and paraben), a sugar (e.g., sucrose, etc.), a reconstitution accelerator, a negatively charged component (e.g., bile salts, bile acids, glycocholic acid, cholic acid, chenodeoxycholic acid, taurocholic acid, glycochenodeoxycholic acid, taurochenodeoxycholic acid, lithocholic acid, ursodeoxycholic acid, dehydrocholic acid, phospholipids).

Preferably, the preparation of the present invention can be prepared by aseptic preparation, terminal filtration or radiation sterilization, and the obtained preparation is preserved in solution or prepared into lyophilized preparation, and further preferably, the preparation of the present invention is intravenous injection preparation.

In yet another aspect, the sterile lyophilized compositions or formulations of cabazitaxel and albumin of the invention may be used to treat cancer, including but not limited to one or more of prostate cancer, colon cancer, head and neck squamous cell carcinoma, breast cancer, head and neck cancer, pancreatic cancer, bladder cancer, liposarcoma, lung cancer, gastric adenocarcinoma, esophageal cancer, and ovarian cancer.

The dosage of administration of the lyophilized composition or formulation of the present invention may vary depending on the particular method of administration and the particular type of cancer being treated. The dosage is sufficient to produce the desired beneficial effect. The composition may be formulated for single dose administration or multiple dose administration.

The lyophilized compositions or formulations of the present invention may be administered by any route known to those skilled in the art, including intramuscular, intravenous, intradermal, intralesional, intraperitoneal injection, or any other suitable route. Administration may be local, topical or systemic, depending on the site of treatment. The most suitable route in any given case will depend on a number of factors, such as the nature of the disease, the course of the disease, the severity of the disease, etc. Preferably, the pharmaceutical composition or pharmaceutical formulation of the invention is administered by intravenous injection.

The lyophilized composition or formulation of the present invention can also be administered with a second active compound/second therapeutic agent. The lyophilized composition or pharmaceutical formulation of the present invention may be administered simultaneously, sequentially or concurrently with the second active compound/second therapeutic agent. When administered separately, the lyophilized composition or formulation of the present invention and the second active compound or second therapeutic agent can be administered at different dosing frequencies or intervals.

The second active compound/second therapeutic agent useful in the present invention may include, but is not limited to, anthracyclines (e.g., epirubicin), nucleosides (e.g., gemcitabine), and the like.

In order to solve the solubility defect of cabazitaxel, the prior art mostly adopts a mode of a human serum albumin nano preparation. The preparation method of the albumin nano preparation is usually a disulfide bond formation-based technology, namely, firstly, a medicament is dissolved in an organic solvent immiscible with water to obtain an oil phase, albumin is dissolved in an aqueous medium to obtain a water phase, and then the oil phase and the water phase are mixed and homogenized to prepare the nano particles. However, in the method, water-insoluble organic solvents such as chloroform and dichloromethane are required to be used to ensure that a uniform suspension is obtained in the preparation process, so that fine nanoparticles can be prepared, but the organic solvents have high toxicity and are not suitable for being applied to a human body in a large amount for a long time, albumin denaturation can be caused, and the stability of the protein nanoparticle preparation is damaged.

Patent CN102458112A and patent CN103393632A disclose taxane drug albumin nanoparticle compositions, which use a large amount of highly toxic organic solvents such as chloroform/dichloromethane during the preparation. Patent CN103393632A specifically states in the specification that the content of dichloromethane/chloroform needs to be controlled above 90% (v/v) to ensure the preparation of sterile filterable nanoparticles, and states that in order to obtain sterile filterable particles with a diameter of less than 200nm, it is necessary to have an organic solvent immiscible with water in the organic phase, even if dichloromethane is used directly as the organic phase in the examples; the patent CN1515244A explicitly states that only particles prepared with a very low ethanol fraction (less than 3%, V/V) can be sterile filtered.

In addition, the prior art uses albumin at high concentrations in order to obtain stable carbaalbumin nanoformulations. Patent CN103393632A teaches that the concentration of the aqueous albumin solution is 5-25% (w/v), preferably in the range of 10% -20% (w/v), i.e. in the concentration range of 50-250mg/ml, and patent CN103393632A also mentions that the protein acts as a dispersing, stabilizing microparticle in the nanoparticles, and the use of the protein in a small amount does not produce stable albumin nanoparticles. Because high concentration human serum albumin can lead to adverse reactions such as allergy, body fluid retention and the like, and during albumin collection, the processes of blood collection, storage and the like can be polluted, and potential safety hazards of blood products can also exist. Therefore, the use of albumin in large quantities is not only disadvantageous to clinical safety, but also increases production costs. The sterile freeze-drying composition of cabazitaxel and albumin and the preparation method thereof have the following beneficial effects:

1. the composition of cabazitaxel and albumin does not need to use surfactants such as Tween-80 and the like. The pharmaceutical composition of the invention avoids adding surfactants such as Tween-80 and the like by adjusting the prescription and the process parameters, can also prepare a stable composition, and overcomes the serious allergic defect caused by the surface activity.

2. The composition of cabazitaxel and albumin only uses alcohol organic solvent in the preparation process, avoids using organic solvents such as dichloromethane/chloroform and the like, and overcomes the defect that solvents with high toxicity and water immiscibility need to be added in the prior art.

3. The composition of cabazitaxel and albumin reduces the using concentration of albumin, ensures the safety of clinical medication and reduces the production cost.

4. The composition of cabazitaxel and albumin of the invention has an average particle size of less than 200nm and excellent stability.

5. When the composition of cabazitaxel and albumin is used for preparing a freeze-dried preparation, a freeze-drying protective agent and a protein stabilizing agent do not need to be added, and a qualified freeze-dried preparation can be prepared.

Drawings

Fig. 1 shows a particle size distribution diagram of cabazitaxel albumin nanoparticles of example 1.

Fig. 2 shows a particle size distribution diagram of cabazitaxel albumin nanoparticles of example 2.

Fig. 3 shows the particle size distribution profile of cabazitaxel albumin nanoparticles of example 3.

Fig. 4 shows the particle size distribution profile of cabazitaxel albumin nanoparticles of example 4.

Fig. 5 shows a particle size distribution diagram of cabazitaxel albumin nanoparticles of example 5.

Fig. 6 shows the particle size distribution profile of cabazitaxel albumin nanoparticles of example 6.

Fig. 7 shows the particle size distribution profile of cabazitaxel albumin nanoparticles of example 7.

Fig. 8 shows zeta potential maps of cabazitaxel albumin nanoparticles of example 1.

Detailed Description

The present invention will be further illustrated with reference to the following examples, which are not intended to limit the scope of the present invention in any way.

The rpm in the present invention is a unit of rotation speed, i.e., rpm; psi pressure, i.e., pounds per square inch.

The cabazitaxel derivative provided by the invention includes, but is not limited to, compounds with structures similar to cabazitaxel, or compounds in the same general chemical class with cabazitaxel, cabazitaxel analogues, or pharmaceutically acceptable salts of cabazitaxel or analogues thereof, and may also include pharmaceutically acceptable salts of the compounds.

Measurement of average particle diameter of the present invention: adding water for injection or 0.9% sodium chloride solution into sample to be measured, dispersing uniformly to obtain sample stock solution, taking appropriate amount of sample stock solution, diluting with water to semitransparent (or diluting with water by 5-30 times), and measuring average particle diameter with nanometer particle size analyzer (Zetasizer Nano ZS, Malvern instruments Co., Ltd.).

Measurement of Zeta potential of the present invention: the sample to be measured was added with water for injection or 0.9% sodium chloride solution and dispersed uniformly, and the Zeta potential was measured using a nanometer particle sizer (Zetasizer Nano ZS, malvern instruments ltd).

Example 1

Dissolving 300mg of cabazitaxel in 30ml of ethanol to form an organic phase, taking 400ml of human serum albumin aqueous solution (15mg/ml) as a water phase, mixing the water phase with the organic phase, shearing the mixture in a high-speed shearing machine at the rotating speed of 10000rpm, transferring the mixture into a high-pressure homogenizer, homogenizing the mixture at the pressure of 35000psi for 5 times of circulation to prepare suspension, wherein the average particle size of the suspension is 119.8nm and the pH value is 6.7, filtering and sterilizing the suspension through a sterile filter head of 0.22 mu m, directly freezing and drying the suspension for 60 hours to obtain a freeze-dried composition, measuring the zeta potential of the freeze-dried composition by taking physiological saline as a redissolving medium to obtain-5.7 mv, carrying out particle size analysis by using a Malvern particle size tester to obtain the average particle size of 120.3nm, standing the mixture at room temperature, and detecting the cabazitaxel content by High Performance Liquid Chromatography (HPLC) to find that the cabazitaxel content is not changed obviously, the stability is greater than 72 hours.

Example 2

Dissolving 300mg of cabazitaxel in 15ml of ethanol to form an organic phase, taking 500ml of human serum albumin aqueous solution (6mg/ml) as an aqueous phase, mixing the aqueous phase and the organic phase, placing the mixture into a high-speed shearing machine to shear at the rotating speed of 7000rpm, transferring the mixture into a high-pressure homogenizer, homogenizing at 30000psi for 5 cycles to obtain a suspension, the average particle diameter of the particles is 134.4nm, pH is 6.5, filtering the suspension through a sterile filter head with the diameter of 0.22 μm for sterilization, directly freeze-drying for 60h to obtain a freeze-dried composition, freeze-drying the composition after completion of freeze-drying, using normal saline as redissolving medium, measuring zeta potential to-3.4 mv, using Malvern particle size analyzer to make particle size analysis, measuring average particle size to be 135.4nm, standing at room temperature, HPLC detection shows that the content of cabazitaxel is not obviously changed, and the stability is more than 72 hours.

Example 3

Dissolving 300mg of cabazitaxel in 10ml of ethanol to form an organic phase, taking 180ml of human serum albumin aqueous solution (20mg/ml) as a water phase, mixing the water phase and the organic phase, placing the mixture into a high-speed shearing machine, shearing the mixture at the rotating speed of 15000rpm, transferring the mixture into a high-pressure homogenizer, homogenizing under 40000psi pressure for 6 cycles to obtain suspension, the average particle diameter of the particles is 164.4nm, pH is 7.0, filtering the suspension with sterile filter head of 0.22 μm for sterilization, directly freeze-drying for 60 hr to obtain lyophilized composition, freeze-drying the lyophilized composition, using normal saline as redissolving medium, measuring zeta potential to-7.5 mv, using Malvern particle size analyzer to analyze particle size, measuring average particle size to be 165.7nm, standing at room temperature, HPLC detection shows that the content of cabazitaxel is not obviously changed, and the stability is more than 72 hours.

Example 4

Dissolving 300mg of cabazitaxel in 6.7ml of ethanol to form an organic phase, taking 1700ml of human serum albumin aqueous solution (3mg/ml) as a water phase, mixing the water phase and the organic phase, placing the mixture into a high-speed shearing machine to shear at the rotating speed of 12000rpm, transferring the mixture into a high-pressure homogenizer, homogenizing at 30000psi for 6 cycles to obtain a suspension, the average particle diameter of the particles is 184.1nm, pH is 7.5, filtering the suspension through a sterile filter head with the diameter of 0.22 μm for sterilization, directly freeze-drying for 60h to obtain a freeze-dried composition, freeze-drying the composition after completion of freeze-drying, using normal saline as redissolving medium, measuring zeta potential to-4.6 mv, using Malvern particle size analyzer to analyze particle size, measuring average particle size to be 185.1nm, standing at room temperature, HPLC detection shows that the content of cabazitaxel is not obviously changed, and the stability is more than 72 hours.

Example 5

Dissolving 300mg of cabazitaxel in 15ml of ethanol to form an organic phase, taking 1125ml of human serum albumin aqueous solution (4mg/ml) as an aqueous phase, mixing the aqueous phase and the organic phase, placing the mixture into a high-speed shearing machine to shear at the rotating speed of 7000rpm, transferring the mixture into a high-pressure homogenizer, homogenizing under 40000psi pressure for 6 cycles to obtain suspension, the average particle diameter of the particles is 162.1nm, pH is 6.2, filtering the suspension with sterile filter head of 0.22 μm for sterilization, directly freeze-drying for 60 hr to obtain lyophilized composition, freeze-drying the lyophilized composition, using normal saline as redissolving medium, measuring zeta potential to-2.8 mv, using Malvern particle size analyzer to analyze particle size, measuring average particle size to be 163.6nm, standing at room temperature, HPLC detection shows that the content of cabazitaxel is not obviously changed, and the stability is more than 72 hours.

Example 6

Dissolving 300mg of cabazitaxel in 15ml of ethylene glycol to form an organic phase, taking 150ml of human serum albumin aqueous solution (3mg/ml) as an aqueous phase, mixing the aqueous phase and the organic phase, placing the mixture into a high-speed shearing machine to shear at the rotating speed of 7000rpm, transferring the mixture into a high-pressure homogenizer, homogenizing under 40000psi pressure for 5 cycles to obtain suspension, the average particle diameter of the particles is 143.5nm, pH is 6.1, filtering the suspension with sterile filter head of 0.22 μm, directly lyophilizing for 60h to obtain lyophilized composition, lyophilizing, using normal saline as redissolving medium, measuring zeta potential to-8.3 mv, using Malvern particle size analyzer to analyze particle size, measuring average particle size to 143.6nm, standing at room temperature, HPLC detection shows that the content of cabazitaxel is not obviously changed, and the stability is more than 72 hours.

Example 7

Dissolving 300mg of cabazitaxel in 10ml of methanol to form an organic phase, taking 75ml of human serum albumin aqueous solution (10mg/ml) as a water phase, mixing the water phase and the organic phase, placing the mixture into a high-speed shearing machine to shear at the rotating speed of 10000rpm, transferring the mixture into a high-pressure homogenizer, homogenizing under 40000psi pressure for 5 cycles to obtain suspension, the average particle diameter of the particles is 168.7nm, pH is 6.8, filtering the suspension with 0.22 μm sterile filter head, directly lyophilizing for 60h to obtain lyophilized composition, lyophilizing the lyophilized composition, using normal saline as redissolving medium, measuring zeta potential to-6.1 mv, using Malvern particle size analyzer to analyze particle size, measuring average particle size to be 169.0nm, standing at room temperature, HPLC detection shows that the content of cabazitaxel is not obviously changed, and the stability is more than 72 hours.

Test example 1

The lyophilized compositions of examples 1 to 7 were reconstituted with physiological saline and the average particle size and Zeta potential were measured at different time points, respectively, and the results are shown in the following table

The data show that the cabazitaxel preparation has excellent stability, and the particle size and the Zeta potential measured in 72 hours after the cabazitaxel preparation is redissolved by normal saline do not change obviously.

Test example 2

According to the preparation method of example 3, cabazitaxel albumin freeze-dried compositions are prepared by adopting different organic solvents, after the compositions are re-dissolved by physiological saline, the particle sizes of the compositions are respectively measured at different time points, and the influence of the different organic solvents on the average particle size and the stability of cabazitaxel albumin nano-formulations is examined, and the results are shown in the following table:

from the above data, the particle size stability of the cabazitaxel albumin lyophilized composition prepared by using organic solvents such as "ethyl acetate", "dichloromethane", "acetone", "ethanol/chloroform", "ethanol/dichloromethane" was less than 72 hours.

Test example 3

According to the preparation method of example 1, cabazitaxel albumin freeze-dried compositions are prepared by adopting different albumin concentrations, after the compositions are re-dissolved by normal saline, the particle sizes of the compositions are respectively measured at different time points, and the influence of the different albumin concentrations on the average particle size and the stability of cabazitaxel albumin nano-preparations is examined, and the results are shown in the following table:

from the data, when the albumin concentration is too low, the nano-particles cannot be formed, and when the albumin concentration is too high, the particle size stability of the prepared freeze-dried composition is greatly influenced.

Test example 4

The lyophilized compositions prepared in examples 1 to 7 were subjected to HPLC to examine the content of cabazitaxel as a main drug and the change of impurities after standing for various periods

The data show that the cabazitaxel preparation has excellent stability, and the content of the main drug and the content of impurities do not change obviously within 72 hours.

Test example 5

Examination of the Effect of adding/not adding lyoprotectants and protein stabilizers on the Properties of the compositions of the present invention

Homogenizing according to the preparation method of example 1 to obtain a dispersion suspension, filtering and sterilizing the dispersion suspension, adding 5% by mass volume percent of mannitol (W: V) and directly freeze-drying for 60 hours, and after redissolving by normal saline, measuring that the average diameter of cabazitaxel particles is 121.5nm, the Zeta potential is-5.7 mv, the content of main drugs is 99.5% and the content of impurities is 0.08%;

the freeze-dried composition obtained by directly freeze-drying for 60 hours according to the preparation method of example 1 takes physiological saline as a redissolving medium, and the average particle size of cabazitaxel particles is measured to be 120.3nm, the zeta potential is measured to be-5.7 mv, the content of the main drug is 99.9 percent, and the content of impurities is measured to be 0.07 percent.

From the data of this test example, it can be seen that: in the freeze-drying step of the composition, compared with the freeze-drying protective agent and the protein stabilizing agent, the property of the composition without the freeze-drying protective agent and the protein stabilizing agent is basically unchanged. Therefore, the composition does not need to add a freeze-drying protective agent and a protein stabilizing agent in the freeze-drying step, reduces the production cost of the medicament, and ensures the safety of clinical medication.

Test example 6

Examination of the Effect of compositions other than those of the present invention on Properties without addition of lyoprotectants/stabilizers

300mg of cabazitaxel is dissolved in 1.35ml of trichloromethane and 0.15ml of absolute ethanol, the organic phase is added into 28.5ml (9.47%, w/v) of human serum albumin aqueous solution, the mixture is sheared and homogenized for 3 minutes under the condition of low rotation speed to form coarse emulsion, the coarse emulsion is transferred into a high-pressure homogenizer, and the coarse emulsion is homogenized for 7 cycles under the condition of 3,000-30,000 psi pressure. The resulting solution was transferred to a rotary evaporator and evaporated at 40 ℃ under reduced pressure (10-15KPa) for 30 minutes to remove the organic solvent. The resulting dispersion suspension was translucent and passed through a 0.22 μm microfiltration membrane filter with cabazitaxel particles having an average diameter of 153 nm.

Freeze-drying:

A. directly freeze-drying the dispersed suspension for 48 hours, and re-dissolving the dispersed suspension in normal saline to obtain cabazitaxel particles with the average diameter of 155nm, the zeta potential of-21.4 mv, the content of main drugs of 90.4 percent and the content of impurities of 8.4 percent;

B. directly freeze-drying the dispersion suspension for 60 hours, and after redissolving by normal saline, measuring that the average diameter of cabazitaxel particles is 152nm, the zeta potential is-18.7 mv, the content of main drugs is 88.7 percent, and the content of impurities is 11.2 percent;

according to the data of the test example, the cabazitaxel albumin composition is directly freeze-dried to obtain a product without using a freeze-drying protective agent or a stabilizing agent, and the content of the main drug and the content of impurities are obviously inferior to those of the cabazitaxel albumin composition.

All documents mentioned in this application are incorporated by reference into this application and each is incorporated by reference herein in its entirety. Furthermore, it should be understood that various changes and modifications can be made by those skilled in the art after reading the above disclosure, and equivalents may fall within the scope of the invention as defined by the claims.

Claims

1. A sterile lyophilized composition of cabazitaxel and albumin, characterized by: the composition is a freeze-drying nanoparticle system consisting of cabazitaxel and albumin, the mass ratio of the cabazitaxel to the albumin is 1: 10-30, the pH is 6.0-7.5, the average particle size is not more than 200nm, and the zeta potential is-2-9 mv, wherein a freeze-drying protective agent and a protein stabilizing agent are not required to be added in the freeze-drying process;

the preparation method of the sterile freeze-drying composition of cabazitaxel and albumin comprises the following steps:

1) dissolving cabazitaxel in an organic solvent to form an organic phase; the organic solvent is selected from alcohol solvents, and the concentration of cabazitaxel in the organic phase is 10-45 mg/ml; the alcohol solvent is selected from methanol, ethanol, glycol and isopropanol;

3) mixing the organic phase obtained in the step 1) and the water phase obtained in the step 2), shearing at a high speed, and homogenizing to obtain a suspension with the average particle size of not more than 200 nm;

2. The sterile lyophilized composition of cabazitaxel and albumin according to claim 1, wherein: the albumin has a sulfhydryl group or a disulfide bond.

3. The sterile lyophilized composition of cabazitaxel and albumin according to claim 1, wherein: the albumin is human serum albumin.

4. The sterile lyophilized composition of cabazitaxel and albumin according to claim 1, wherein: the mass ratio of the cabazitaxel to the albumin is 1: 10-20; the pH value is 6.5-7.0; the zeta potential is-5 to-8 mv.

5. The sterile lyophilized composition of cabazitaxel and albumin according to claim 4, wherein: the mass ratio of the cabazitaxel to the albumin is 1: 12-17.

6. A method of preparing a sterile lyophilized composition of cabazitaxel and albumin according to claim 1, comprising the steps of:

1) dissolving cabazitaxel in an organic solvent to form an organic phase; the organic solvent is selected from alcohol solvents, and the concentration of cabazitaxel in the organic phase is 10-45 mg/ml;

7. The method of preparing a sterile lyophilized composition of cabazitaxel and albumin according to claim 6, wherein: the alcohol solvent in the step 1) is ethanol.

8. Method for the preparation of sterile lyophilized compositions of cabazitaxel and albumin according to claim 6 or 7, characterized in that: the concentration of albumin in the step 2) is 3-15 mg/ml; the concentration of cabazitaxel in the step 3) and the step 4) is 0.1-6 mg/ml; and (4) carrying out ultrafiltration concentration to 2-20 times.

9. The method of preparing a sterile lyophilized composition of cabazitaxel and albumin according to claim 8, wherein: the concentration of cabazitaxel in the step 3) and the step 4) is 0.4-4 mg/ml.

10. Method for the preparation of sterile lyophilized compositions of cabazitaxel and albumin according to claim 6 or 7, characterized in that: the high-speed shearing speed is 7000-15000 rpm; the homogenizing pressure is 30000-40000 psi; and repeating the homogenization step for 5-6 cycles.

11. The method of preparing a sterile lyophilized composition of cabazitaxel and albumin according to claim 8, wherein: the high-speed shearing speed is 7000-15000 rpm; the homogenizing pressure is 30000-40000 psi; and repeating the homogenization step for 5-6 cycles.

12. A formulation comprising a sterile lyophilized composition of cabazitaxel and albumin according to any one of claims 1 to 5, and a pharmaceutically acceptable carrier and/or adjuvant.

13. A formulation comprising a sterile lyophilized composition of cabazitaxel and albumin prepared according to the process of any one of claims 6 to 11, and a pharmaceutically acceptable carrier and/or adjuvant.

14. The formulation according to claim 12 or 13, wherein the formulation is prepared by aseptic preparation, terminal filtration or radiation sterilization, and the resulting formulation is preserved in solution or prepared as a lyophilized formulation.

15. The formulation of claim 14, wherein the formulation is an intravenous formulation.

16. Use of a sterile lyophilized composition of cabazitaxel and albumin according to any one of claims 1 to 5 or a formulation according to any one of claims 12 to 15 in the manufacture of a medicament for the treatment of a cancer selected from prostate cancer, colon cancer, breast cancer, head and neck cancer, pancreatic cancer, bladder cancer, liposarcoma, lung cancer, gastric adenocarcinoma, esophageal cancer and ovarian cancer.

17. The use according to claim 16, wherein the cancer of the head and neck is a squamous cell carcinoma of the head and neck.