A micelle containing a taxane drug, a preparation method and application thereof

Abstract

The application discloses a micelle containing a taxane drug, a preparation method and application thereof. The application provides a micelle containing substance A, which comprises the following components in parts by weight: 1 part of substance A and 5-25 parts of polyethylene glycol derivatized phospholipid; the substance A is docetaxel and / or cabazitaxel; and the polyethylene glycol derivatized phospholipid comprises a polyethylene glycol part and a phospholipid part, wherein the phospholipid part is di(C 12 ‑C 24 phosphatidylethanolamine. The micelle containing the taxane drug can contain a therapeutically effective amount of the taxane drug, the drug loading capacity is as high as 6.25%-9.09%, the stability is good, and the problems in clinical application can be effectively solved.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of medicine, and particularly relates to a micelle containing a taxane drug, a preparation method and application thereof. BACKGROUND

[0002] Taxane drugs are a large class of natural drugs obtained from plants and belong to diterpenes in structural classification. Cabazitaxel and docetaxel both belong to taxane drugs and are liposoluble compounds. The poor water solubility, short half-life and high toxicity of the drugs limit their clinical application. At present, the marketed docetaxel injection and cabazitaxel injection are both prepared by dissolving the drug in Tween-80. In clinical application, the drug needs to be diluted with a special injection solvent, and the operation requires strict requirements. Special infusion devices are also required for infusion. The use method is complicated, and the preparation contains a large amount of Tween-80, which is very easy to cause hemolysis and allergic reactions. Therefore, it is necessary to take dexamethasone and other drugs in advance for prevention. The clinical medication is not convenient, and the drug safety is low (Annals of Allergy, Asthma and Immunology, 2005, 95(6): 593-599; Journal of Investigational Allergology and Clinical Immunology, 2016, 26, 394-396). So far, this problem has not been well solved.

[0003] There are liposomes containing taxane drugs in the prior art, but they also have many shortcomings. For example, the preparation process of the liposomes is complex, requires the compounding of multiple lipid components (at least two lipid components), and the particle size control requires special equipment and devices; the liposomes are prone to flocculation during storage, etc.

[0004] The encapsulation efficiency of cabazitaxel liposomes prepared by Jiangsu University Zhang Li et al. is only about 86%, and the particle size changes greatly before and after freeze-drying, from 68 nm to 108 nm, which increases the instability of the liposomes after reconstitution. (Zhang Li. Preparation and pharmacokinetic study of cabazitaxel lyophilized liposomes [D]. Jiangsu: Jiangsu University, 2017.)

[0005] The addition of a certain proportion of polyethylene glycol derivatized phospholipid in the general liposome prescription can prepare into long-circulating liposomes. The long chain of PEG forms a steric hindrance and hydrophilic protective layer on the surface of the liposome, which can prevent the recognition and uptake of the liposome by phagocytes, increase its circulation time in the blood, and endow this type of liposome with long-circulating properties. However, when the mass ratio of polyethylene glycol derivatized phospholipid and phospholipid exceeds a certain value, the drug encapsulation efficiency and stability will decrease significantly, because when the content of polyethylene glycol derivatized phospholipid is high, it is difficult to insert into the phospholipid bilayer of the liposome, and it is easy to form mixed micelles with phospholipid, which affects the stability of the liposome. (Colloids and Surfaces B: Biointerfaces 155 (2017): 266-275.)

[0006] Polymer nanomicelles are a drug delivery system developed in recent years for poorly soluble drugs, which have a core-shell structure, with the core being the hydrophobic part and the shell being the hydrophilic part. Polymer nanomicelles can load poorly soluble drugs at the core-shell interface to achieve solubilization of poorly soluble drugs. Compared with commonly used solubilizers such as Tween-80 and polyoxyethylated castor oil, polymer micelles are often made of biodegradable materials, which are safe, and therefore have good application prospects as carriers for poorly soluble drugs (Journal of Controlled Release, 2001, 73, 137-172; Pharmaceutical Research, 2007, 24(1), 1-16). At present, there are many nanofactories of taxane drugs under research (CN103990135A; CN107625730A; CN104758256A), which have solved the problems of poor water solubility, adverse reactions, and low bioavailability of taxane drugs to some extent. However, the current nanofactories of taxane drugs all have the problems of low drug loading, low encapsulation efficiency, and poor stability (CN103990135A; CN107625730A; CN104758256A; CN102293736A; CN101732234A; CN101804021A; CN101829046A). The drug loading of existing nanofactories is generally less than 5%, and the encapsulation efficiency is less than 90%, which leads to the introduction of a large amount of carrier materials in the preparation. Since many nanomaterials have not been safety verified in clinical practice, and even have the risk of generating toxic substances during degradation in the human body, which will limit their use in clinical practice (Journal of Controlled Release, 2009, 133, 11-17; International Journal of Pharmaceutics, 2014, 464(1-2):178-184.); On the other hand, most nanofactories of poorly soluble drugs have the problem of poor stability, and the encapsulation efficiency will decrease in a very short time, leading to drug leakage. Especially for taxane drugs, when the encapsulation efficiency decreases to less than 95%, insoluble particles will appear in the drug solution, affecting the quality of the preparation and making it impossible to be truly applied in clinical practice (CN103990135A, CN107625730A, CN104758256A); At the same time, many nanofactories have a very complex preparation process before use, and some even need to be heated and stirred, which brings many risks to clinical application (CN101972480A; Drug Delivery and Translational Research, 2018, 5, 1365-1379).

[0007] PEG-DSPE, as a micellar drug delivery system, has a series of unique advantages, and the PEG-DSPE polymer molecule is a FDA-approved pharmaceutical excipient, which is safe (Journal of Controlled Release, 2013, 171, 133-142; Pharmaceutical Research, 2012, 29, 1977-1989; Journal of the National Cancer Institute, 2007, 99, 1004-1015). Our previous studies have shown that some water-soluble small molecule drugs can be successfully loaded into the PEG-DSPE nanomicelles formed by a simple one-step self-assembly in aqueous solution (Pharmaceutical Research, 2010, 27(2), 361-370), and these drug-loaded micelles can unload their effective drug payloads onto the cell membrane by disassembling and inserting the cell membrane, thereby increasing the membrane fluidity and accelerating the intracellular transport of the drug (Journal of Controlled Release, 2012, 160, 637-651). At the same time, we recently found that PEG-DSPE nanomicelles have the function of natural molecular chaperones, and the hydrophilic nanocage formed by the outer shell can accommodate the A chain and B chain of insulin and promote the correct folding of the two chains and prevent the aggregation of insulin. According to the spatial structure of the nanocage formed, PEG-DSPE nanomicelles can accommodate protein molecules with a molecular weight of less than 20KD (Biomaterials, 2016, 77, 139-148). Although PEG-DSPE has excellent drug loading capacity, it has a certain structure selectivity for the loaded drugs, such as doxorubicin hydrochloride, vinorelbine tartrate, and vincristine sulfate, which can be successfully loaded into PEG-DSPE micelles with a high encapsulation efficiency of 99.9% (CN1840193A; CN101322681A; CN1739525A; CN101138545A). However, PEG-DSPE micelles do not have the ability to load all small molecule drugs, such as gemcitabine hydrochloride and paclitaxel, which cannot be successfully and stably loaded into PEG-DSPE micelles, and cannot achieve the purpose of clinical drug use (Pharmaceutical Research, 2010, 27(2), 361-370; Philosophical Transactions of The Royal Society A, 20120309, 371; Molecular Cancer Therapeutics, 2014, 13(12), 2864-2875). SUMMARY

[0008] The technical problem to be solved by the present application is to overcome the problem that the prior art cannot effectively prepare a taxane nanomicelle preparation with high drug loading, high encapsulation efficiency and good stability; and a micelle containing a taxane drug, a preparation method and application thereof are provided. Low drug loading, low encapsulation efficiency and poor stability are the main factors affecting the clinical application of nanomedicine preparations. The taxane nanomicelle provided by the present application contains a therapeutically effective amount of taxane; the drug loading is as high as 6.25% to 9.09%, the encapsulation efficiency is 98% to 100%, and the stability (and encapsulation efficiency of 98% to 100%) of the nanomicelle after reconstitution is greater than 24 hours; which can effectively solve the problems in clinical application. At the same time, the taxane nanomicelle provided by the present application has high drug loading, high encapsulation efficiency and good stability, is a kinetic stable system, is simple to prepare clinically, does not need to use special infusion devices, and has a targeting effect in vivo, can increase the distribution of drugs in tumor tissues, improve the efficacy and reduce toxicity.

[0009] The present application solves the above technical problems by the following technical solutions.

[0010] The present application provides a micelle containing substance A, which comprises the following components in parts by weight: 1 part of substance A and 5 to 25 parts of polyethylene glycol derivatized phospholipid; the substance A is docetaxel and / or cabazitaxel; the polyethylene glycol derivatized phospholipid comprises a polyethylene glycol moiety and a phospholipid moiety, and the phospholipid moiety is one or more of di(C 12 -C 24 fatty acyl)phosphatidylethanolamine.

[0011] In an embodiment of the present application, the phospholipid moiety of the polyethylene glycol derivatized phospholipid can be one or more of di(C 18 , abbreviated as DSPE), dipalmitoylphosphatidylethanolamine (C 16 , abbreviated as DPPE), dimyristoylphosphatidylethanolamine (C 14 , abbreviated as DMPE), and dioleoylphosphatidylethanolamine (C 18 , abbreviated as DOPE).

[0012] The polyethylene glycol derivatized phospholipid has a polyethylene glycol molecular weight in the range of 1000 to 5000, for example, a polyethylene glycol molecular weight of 2000.

[0013] In some embodiments of the present application, the polyethylene glycol derivatized phospholipid can be polyethylene glycol 1000 distearoylphosphatidylethanolamine (PEG1000-DSPE), polyethylene glycol 2000 distearoylphosphatidylethanolamine (PEG2000-DSPE), polyethylene glycol 5000 distearoylphosphatidylethanolamine (PEG5000-DSPE), polyethylene glycol 2000 dioleoylphosphatidylethanolamine (PEG2000-DOPE), polyethylene glycol 2000 dipalmitoylphosphatidyl ethanolamine (PEG2000-DPPE), or polyethylene glycol 2000 dimyristoylphosphatidylethanolamine (PEG2000-DMPE); since the PEG-DSPE polymer molecule is a FDA-approved pharmaceutical excipient, it is safe, and thus, PEG2000-DSPE is preferred.

[0014] In some embodiments of the present application, by weight, it comprises the following components: 1 part of substance A and 10-15 parts of polyethylene glycol derivatized phospholipid.

[0015] In some embodiments of the present application, the micelles of the substance A are composed of the substance A and the polyethylene glycol derivatized phospholipid.

[0016] In some embodiments of the present application, by weight, it comprises the following components: 1 part of docetaxel and 5 or 15 parts of polyethylene glycol derivatized phospholipid (e.g., PEG2000-DSPE). For example, it is composed of 1 part of docetaxel and 5 or 15 parts of the polyethylene glycol derivatized phospholipid (e.g., PEG2000-DSPE). The drug loading capacity can be as high as 6.25%.

[0017] In some embodiments of the present application, by weight, it comprises the following components: 1 part of cabazitaxel and 5, 10 or 15 parts of polyethylene glycol derivatized phospholipid (e.g., PEG1000-DSPE, PEG2000-DSPE, PEG5000-DSPE, PEG2000-DOPE, PEG2000-DPPE, PEG2000-DMPE). For example, it is composed of 1 part of cabazitaxel and 5, 10 or 15 parts of polyethylene glycol 2000 distearoylphosphatidylethanolamine, or 1 part of cabazitaxel and 10 parts of polyethylene glycol 1000 distearoylphosphatidylethanolamine, polyethylene glycol 5000 distearoylphosphatidylethanolamine, polyethylene glycol 2000 dioleoylphosphatidylethanolamine, polyethylene glycol 2000 dipalmitoylphosphatidyl ethanolamine, or polyethylene glycol 2000 dimyristoylphosphatidylethanolamine). The drug loading capacity can be as high as 9.09%.

[0018] In the present application, the micelles containing substance A are spherical with core-shell structure; wherein the inner core is a phospholipid layer, the outer shell is a polyethylene glycol (PEG) layer, and the substance A is distributed in the polyethylene glycol derivatized phospholipid micelles (i.e. wrapped in the micelles inside the core-shell boundary layer).

[0019] In some embodiments of the present application, the particle size of the micelles can range from 5 to 50 nm, preferably from 10 to 20 nm.

[0020] In some embodiments of the present application, the polydispersity (PDI) of the micelles is less than 0.3, and the particle size distribution is relatively uniform; preferably less than 0.2, for example when the substance A is cabazitaxel, the polydispersity (PDI) is 0.100-0.140. When the substance A is docetaxel, the polydispersity is 0.100-0.180.

[0021] In some embodiments of the present application, when the polyethylene glycol derivatized phospholipid is PEG2000-DSPE, the average diameter of the micelles is 10-15 nm (e.g. 12 nm).

[0022] In some embodiments of the present application, when the polyethylene glycol derivatized phospholipid is PEG2000-DSPE, the aggregation number of the micelles is about 90.

[0023] Another object of the present application is to provide a preparation method of a substance containing a taxane drug, which comprises the following steps:

[0024] Step 1: removing the organic solvent in the solution to obtain a polymeric lipid film containing a taxane drug; wherein the solution is composed of 1 part of a taxane drug, 5-25 parts of a polyethylene glycol derivatized phospholipid and the organic solvent; the taxane drug is docetaxel and / or cabazitaxel; the polyethylene glycol derivatized phospholipid comprises a polyethylene glycol moiety and a phospholipid moiety, and the phospholipid moiety is one or more of di(C 12 -C 24 fatty acyl)phosphatidylethanolamine;

[0025] Step 2: hydrating the polymeric lipid film obtained in step 1 in an aqueous solvent to obtain a substance containing a taxane drug.

[0026] In some embodiments of the present application, the phospholipid moiety in the polyethylene glycol derivatized phospholipid can be one or more of di(C

[0027] The polyethylene glycol derivatized phospholipid has a polyethylene glycol molecular weight ranging from 1000 to 5000, for example, a polyethylene glycol molecular weight of 2000.

[0028] In some embodiments of the present application, the polyethylene glycol derivatized phospholipid can be polyethylene glycol 1000 distearoyl phosphatidyl ethanolamine (PEG1000-DSPE), polyethylene glycol 2000 distearoyl phosphatidyl ethanolamine (PEG2000-DSPE), polyethylene glycol 5000 distearoyl phosphatidyl ethanolamine (PEG5000-DSPE), polyethylene glycol 2000 dioleoyl phosphatidyl ethanolamine (PEG2000-DOPE), polyethylene glycol 2000 dipalmitoyl phosphatidyl ethanolamine (PEG2000-DPPE), or polyethylene glycol 2000 dimyristoyl phosphatidyl ethanolamine (PEG2000-DMPE). Since the PEG-DSPE polymer molecule is a FDA-approved pharmaceutical excipient, it is safe, and thus, PEG2000-DSPE is preferred.

[0029] In some embodiments of the present application, the solution comprises, by weight fraction, 1 part of a taxane drug and 10 to 15 parts of a polyethylene glycol derivatized phospholipid.

[0030] In some embodiments of the present application, the solution comprises, by weight fraction, 1 part of docetaxel and 5 or 15 parts of a polyethylene glycol derivatized phospholipid (for example, PEG2000-DSPE).

[0031] In some embodiments of the present application, the solution comprises, by weight fraction, 1 part of cabazitaxel and 5, 10 or 15 parts of a polyethylene glycol derivatized phospholipid (for example, consisting of 1 part of cabazitaxel and 5, 10 or 15 parts of polyethylene glycol 2000 distearoyl phosphatidyl ethanolamine, or consisting of 1 part of cabazitaxel and 10 parts of polyethylene glycol 1000 distearoyl phosphatidyl ethanolamine, polyethylene glycol 5000 distearoyl phosphatidyl ethanolamine, polyethylene glycol 2000 dioleoyl phosphatidyl ethanolamine, polyethylene glycol 2000 dipalmitoyl phosphatidyl ethanolamine, or polyethylene glycol 2000 dimyristoyl phosphatidyl ethanolamine).

[0032] In step 1, the organic solvent can be a conventional organic solvent in the art for such preparation, for example, one or a mixture of at least two of an alcohol solvent (for example, methanol and / or ethanol), a halogenated hydrocarbon solvent (for example, chloroform), and a nitrile solvent (for example, acetonitrile), preferably ethanol and / or acetonitrile, and most preferably ethanol.

[0033] The skilled in the art can understand that the solution of the taxane drug, the polyethylene glycol derivatized phospholipid and the organic solvent in step 1) is to dissolve the drug and the polyethylene glycol derivatized phospholipid in the organic solvent so that they can contact at the molecular level. Therefore, the amount of the organic solvent can not be particularly limited and can be controlled to dissolve the drug and the polyethylene glycol derivatized phospholipid.

[0034] In the present application, the operation of removing the organic solvent can be a conventional operation in the art, for example, removing the organic solvent by a rotary evaporation method under reduced pressure and / or removing the organic solvent under vacuum. In the present application, the rotary evaporation temperature is 30-60°C, preferably 40°C±5°C, and the rotary speed is 10-500 r / min, preferably 60-150 r / min.

[0035] In the present application, the aqueous solvent is selected from one or a mixture of at least two of deionized water, a PBS buffer solution (for example, a 0.01M-0.2M phosphate buffered saline solution), normal saline (a 0.9% sodium chloride aqueous solution), a glucose injection (for example, 5-50 g / mL), and an amino acid injection (for example, 5-10 g / mL), and is preferably deionized water.

[0036] In the present application, the mass / volume ratio of the taxane drug in the aqueous solvent can be 0.5-15 mg / mL, preferably 5-10 mg / mL (for example, 5-6 mg / mL when the taxane drug is docetaxel, and 5-8 mg / mL when the taxane drug is cabazitaxel).

[0037] The hydration operation can be a conventional operation in the art, and in the present application, can be vortex shaking or ultrasonic hydration. The vortex shaking hydration can be performed for 1-3 hours. The ultrasonic hydration time can be 1-60 minutes, preferably 10-30 minutes (for example, 20 minutes), and the ultrasonic intensity can be 0.1-15 w / cm 2 (for example, 10 w / cm 2 ), and the frequency can be 0.1-100 kHz (for example, 40 kHz).

[0038] The hydration temperature can be a conventional temperature in the art, for example, 0-60°C. In the present application, the temperature is preferably 20-40°C (for example, a water bath at 20-40°C), for example, 30-40°C.

[0039] In some embodiments of the present application, the hydration can be performed in the absence or presence of pharmaceutical excipients. Preferably, the hydration is performed in the absence of pharmaceutical excipients.

[0040] In some embodiments of the present application, the step 2 can further comprise a filtration step, which can be a conventional filtration step in the art. For example, the filtration step can comprise the following steps: after hydration (e.g., when the aqueous solvent is deionized water), the mixture is filtered through a nanometer filter (to remove impurities), and the resulting product containing the taxane drug is obtained; or, after hydration (e.g., when the aqueous solvent is deionized water), the mixture of the taxane drug and the pharmaceutical excipient is filtered through a nanometer filter (to remove impurities), and the resulting product containing the taxane drug is obtained; preferably, after hydration in deionized water, the mixture of the taxane drug and the pharmaceutical excipient is filtered through a nanometer filter (to remove impurities), and the resulting product containing the taxane drug is obtained.

[0041] In some embodiments of the present application, the pharmaceutical excipient used in step 2 can be a conventional pharmaceutical excipient used in micelles in the art, such as a freeze-drying excipient (also known as a freeze-drying protective agent). The freeze-drying excipient can be one or more of glucose, mannitol, lactose, trehalose, sucrose, dextran, glycine, fructose, and sorbitol, preferably glucose. The mass ratio of the freeze-drying excipient to the taxane drug can be 0 to 25:1 (e.g., 15.6:1, 20.8:1). The freeze-drying excipient has another function, which can help the freeze-dried powder to be quickly reconstituted, reducing the time required for reconstitution of the freeze-dried powder to be clear.

[0042] In some embodiments of the present application, the nanometer filter can have a pore size of 10-300 nm, preferably a pore size of 220 nm, and the filter material can be a mixed cellulose ester microporous filter, a nylon filter, a polytetrafluoroethylene filter, a polyvinylidene fluoride membrane, a polyether sulfone filter, or a polypropylene filter, preferably a polyvinylidene fluoride membrane or a polyether sulfone filter, and most preferably a polyether sulfone filter.

[0043] In some embodiments of the present application, the following steps can be included: dissolving the taxane drug and the polyethylene glycol-derived phospholipid in the organic solvent (e.g., ethanol), rotary evaporation (e.g., at 30-60°C), removing the organic solvent, forming a uniform polymer lipid film, adding an aqueous solvent (e.g., deionized water), sonicating in a water bath at 20-40°C (e.g., 30-40°C), filtering through a nanometer filter (e.g., 220 nm) to remove impurities, and obtaining the product containing the taxane drug; or, after hydration, mixing with a pharmaceutical excipient, filtering through a nanometer filter (e.g., 220 nm) to remove impurities, and obtaining the product containing the taxane drug.

[0044] In some embodiments of the present application, the paclitaxel-containing substance is composed of the paclitaxel, the polyethylene glycol derivatized phospholipid, and the aqueous solvent; or, when the pharmaceutical excipient is present, the paclitaxel-containing substance is composed of the paclitaxel, the polyethylene glycol derivatized phospholipid, the pharmaceutical excipient, and the aqueous solvent.

[0045] In some embodiments of the present application, the preparation method further comprises a freeze-drying step, which can be performed according to conventional methods.

[0046] In some embodiments of the present application, the paclitaxel-containing substance is a micelle containing the paclitaxel, such as the micelle containing Substance A as described above, which is formed by the following principle: the polyethylene glycol derivatized phospholipid in the paclitaxel-containing polymeric lipid membrane self-assembles into a complex framework in the aqueous solvent as a carrier material through interaction forces after the hydration, and the paclitaxel is wrapped in the micelle formed by the polyethylene glycol derivatized phospholipid.

[0047] In the present application, when the paclitaxel-containing substance is a micelle containing the paclitaxel, the particle size and particle size distribution of the micelle can be reduced by high-pressure homogenization, high-speed homogenization, or ultrasonic technology. The particle size of the micelle can range from 5 nm to 50 nm, preferably from 10 nm to 20 nm. The polydispersity (PDI) of the micelle is less than 0.3, and the particle size distribution is relatively uniform; preferably less than 0.2. For example, when the Substance A is cabazitaxel, the polydispersity (PDI) is 0.100-0.140. When the Substance A is docetaxel, the polydispersity is 0.100-0.180. The average diameter of the micelle can be 10-15 nm. When the polyethylene glycol derivatized phospholipid is PEG2000-DSPE, the aggregation number of the micelle can be 90. The encapsulation efficiency of the paclitaxel can be greater than 98% (e.g., 100%). The room temperature stability of the docetaxel nanomicelle is not less than 4 hours. The room temperature stability of the cabazitaxel nanomicelle is not less than 8 hours.

[0048] The present application also provides a paclitaxel-containing substance prepared according to the preparation method as described above.

[0049] As understood by those skilled in the art, the "substantially free" of organic solvent in the substance containing the taxane drug; "substantially free" as used herein means no or insignificant amount (i.e. an amount that has no measurable effect on the physical properties of the substance containing the taxane drug). For example, the organic solvent can be excluded or also included as described above; the organic solvent is the organic solvent that is conventionally used in the preparation process of the present application (e.g. methanol, ethanol, chloroform, acetonitrile or a mixture thereof, for example ethanol and / or acetonitrile, for example ethanol). The organic solvent can be within 2% (weight percentage of organic solvent / drug).

[0050] The present application provides a pharmaceutical composition comprising the micelle containing substance A as described above or the substance containing the taxane drug as described above, and a pharmaceutical excipient.

[0051] The pharmaceutical composition can be a preparation in lyophilized form, injection, oral preparation, etc., which is prepared by a conventional preparation process in the art, such as freeze-drying, spray-drying, evaporation under reduced pressure, etc.

[0052] In some embodiments of the present application, the pharmaceutical composition can be in solution form or in lyophilized form as needed, preferably in lyophilized form.

[0053] The pharmaceutical excipient can be a conventional pharmaceutical excipient in the pharmaceutical composition in the art; for example, an aqueous solvent and / or a lyophilized excipient; the aqueous solvent can be one or a mixture of at least two of deionized water, PBS buffer solution (e.g. 0.01M-0.2M phosphate buffered saline solution), physiological saline (0.9% sodium chloride aqueous solution), glucose injection (e.g. 5-50g / mL) and amino acid injection (e.g. 5-10g / mL), preferably deionized water; the lyophilized excipient can be one or more of glucose, mannitol, lactose, trehalose, sucrose, dextran, glycine, fructose and sorbitol, preferably glucose.

[0054] The present application also provides a use of the micelle containing substance A as described above or the substance containing the taxane drug as described above in the preparation of a medicament for treating cancer. The substance A or the taxane drug can be in a therapeutically effective amount. The cancer can be selected from breast cancer, ovarian cancer, non-small cell lung cancer, head and neck cancer, pancreatic cancer, small cell lung cancer, gastric cancer, melanoma, soft tissue sarcoma.

[0055] In order to better understand the content of the present application, some professional terms are explained as follows.

[0056] The term "pharmaceutically acceptable" is used in the sense of those compounds, materials, compositions, and / or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0057] In the present application, "pharmaceutical composition" refers to a preparation of a compound of the present application with a medium conventionally accepted in the art for the delivery of biologically active compounds to mammals (e.g., humans). The medium includes pharmaceutical excipients. The purpose of a pharmaceutical composition is to facilitate administration of the organism, to facilitate absorption of the active ingredient, and to thereby effect biological activity.

[0058] The pharmaceutical excipients can be those widely used in the pharmaceutical production field. The excipients are mainly used to provide a safe, stable and functional pharmaceutical composition, and can also provide a method for the active ingredient to be eluted at a desired rate after the subject is administered, or to facilitate the active ingredient to be effectively absorbed after the subject is administered with the composition. The pharmaceutical excipients can be inert fillers, or provide certain functions, such as stabilizing the overall pH of the composition or preventing degradation of the active ingredient of the composition. The pharmaceutical excipients can include one or more of the following excipients: binders, suspending agents, emulsifiers, diluents, fillers, granulating agents, adhesives, disintegrants, lubricants, anti-adherents, glidants, wetting agents, gelling agents, absorption delaying agents, dissolution inhibitors, enhancers, adsorbents, buffers, chelating agents, preservatives, colorants, flavorants, and sweeteners.

[0059] The pharmaceutical composition of the present application can be prepared according to the disclosure using any method known to those skilled in the art. For example, conventional mixing, dissolving, granulating, emulsifying, dragee-making, encapsulating, entrapping or lyophilizing processes.

[0060] The pharmaceutical composition of the present application can be administered in any form, including injection (intravenous), mucosal, oral (solid and liquid preparations), inhalation, ocular, rectal, topical or parenteral (infusion, injection, implantation, subcutaneous, intravenous, intra-arterial, intramuscular) administration.

[0061] The term "micelle" refers to the ability of amphiphilic molecules to spontaneously aggregate to form micelles when the concentration of the molecules in an aqueous solution exceeds the critical micelle concentration (CMC). The structure of micelles is different from that of liposomes, which do not have the structural characteristics of lipid bilayers. Generally, the structure of micelles is that the hydrophobic part is inward, forming a hydrophobic core, and the hydrophilic part is outward, forming a hydrophilic surface. The particle size of micelles is small, with an average particle size of about 10-50 nm. Therefore, it is not only a thermodynamically stable system, but also a kinetically stable system. In addition, micelle particles are not easy to aggregate and delaminate, and have high loading capacity, i.e., they can load a relatively high amount of drugs at low concentrations.

[0062] The term "micellar aggregation number" is a measure of micelle size, i.e. the number of surfactant molecules or ionic monomers associated into a micelle.

[0063] The term "therapeutically effective amount" means the amount of the substance A or the taxane drug that produces a therapeutic effect. According to the present application, the unit dose (the amount of the drug used per square meter of body surface area) of the substance A or the taxane drug is 5-100 mg / m 2 , preferably 10-20 mg / m 2 , and most preferably 20 mg / m 2 . The dose will be adjusted according to the needs of each particular individual.

[0064] In the present application, the polyethylene glycol derivatized phospholipid is formed by the combination of polyethylene glycol molecules and nitrogen-containing base on the phospholipid molecules through covalent bond. In the structure, the number of carbon atoms in the fatty acid of the phospholipid moiety is 10-24, preferably 12, 14, 16, 18, 20, 22, 24 (e.g. 14, 16, 18), and more preferably 18; the fatty acid chain can be saturated or partially saturated, preferably the fatty acid is one or more of lauric acid, myristic acid, palmitic acid, stearic acid or oleic acid, linoleic acid, eicosanoic acid, behenic acid and lignocerate (tetracosanoic acid).

[0065] The micellar preparation of the present application uses polyethylene glycol derivatized phospholipid as the main matrix, and the polyethylene glycol molecules form a hydrophilic protective layer outside the hydrophobic core containing the drug, which can avoid the contact of the drug with the protein molecules such as enzymes in the blood and protect the nanomicelles from being phagocytosed by the reticuloendothelial system in the body, thus prolonging the retention time of the nanomicelles in the blood circulation; the drug is contained in the hydrophobic core of the micelles, which can protect the drug from the damage of external factors (water, oxygen, light) and greatly improve the stability of the drug during storage; in addition, the drug distribution kinetics in the body is changed, thus enhancing the therapeutic effect and reducing the toxicity.

[0066] On the basis of the common knowledge in the art, the above-mentioned preferred conditions can be combined arbitrarily, i.e. to obtain the preferred examples of the present application.

[0067] The reagents and raw materials used in the present application are commercially available.

[0068] The positive progress effect of the present application is that: 1) the provided taxane micellar preparation selects a safe and simple polymer carrier material, improves the solubility of the taxane and the compliance of the preparation, realizes the drug loading amount with sufficient clinical effect, the drug loading amount can be as high as 6.25% to 9.09%, and the drug encapsulation rate is as high as 98% to 100%, and the drug loading of the taxane can be realized with a lower amount of auxiliary materials.

[0069] 2) The taxane nanopowder preparation of the present application improves the stability, compliance and safety of the drug. The powder preparation can be hydrated into a nanomicelle preparation after dissolution, has good stability, and the room temperature storage stability (encapsulation rate 98% to 100%) is greater than 24 hours, and can be directly used for intravenous injection without special infusion device. Under in vitro conditions, due to the absence of albumin, immunoglobulin, hemoglobin and other related proteins in the body, the stability is higher; when the drug-loaded micelles enter the blood, due to the blood flow rate, blood pressure, and protein interaction, the micelles will gradually dissociate and release drug molecules.

[0070] 3) The average particle size of the taxane micelles is 10-20 nm, which can realize passive targeting of the nanometer preparation to the tumor by using the penetration enhancement and retention effect (EPR effect) of the tumor tissue, promote the selective distribution of the drug in the tumor tissue, and can increase the drug efficacy and reduce the systemic toxic side effects.

[0071] 4) The preparation process of the present application is simple, reproducible, and suitable for industrial production. BRIEF DESCRIPTION OF DRAWINGS

[0072] Figure 1 The effect of different cabazitaxel formulations on the body weight of mice in Example 9.

[0073] Figure 2 The cytotoxic effect of cabazitaxel micelles on MC-38 cells in Example 10. DETAILED DESCRIPTION

[0074] The present application will be further described by way of examples, but the present application is not limited in the scope of the examples. The experimental methods in the following examples are not specified, and are selected according to conventional methods and conditions, or according to the instructions of the commodity.

[0075] Drug-loaded micelle particle size analysis

[0076] The particle size distribution of the drug-loaded micelles was determined by dynamic light scattering (DLS). The sample was diluted to 1 mg / mL with normal saline, 1 mL of the sample was taken, and the particle size distribution and polydispersity PDI of the drug-loaded micelles were measured by a Malvern Zetasizer Nano-ZS light scattering particle size analyzer. The wavelength of the laser beam of the instrument was set to 633 nm, and the angle between the incident light and the scattered light beam was 90°. Each sample was measured for 20 cycles, and the measurement temperature was set to 25°C. The measurement results of each preparation were the average of the measurement results of 3 batches of preparations. The polydispersity (PDI) represents the uniformity of the particle size, and the smaller the polydispersity, the more uniform the particles.

[0077] Drug encapsulation efficiency and drug loading

[0078] 1 mL of the drug-loaded micelle solution was filtered through a 220 nm microporous filter, and the concentration of the drug-loaded micelles in the filtrate was determined by high performance liquid chromatography. The encapsulation efficiency and drug loading were calculated using the following formula.

[0079] Encapsulation efficiency % = C2 / C1 x 100%

[0080] In the formula, C1 is the drug concentration in the drug-loaded micelle solution; C2 is the drug concentration in the drug-loaded micelle solution after filtration through a 220 nm filter (the drug not encapsulated by the micelles is removed in solid form). The drug concentration released from the drug-loaded micelles in the solution was determined by high performance liquid chromatography.

[0081] Drug loading % = (C2 x V) / (C1 x V + M) x 100%

[0082] In the formula, V is the volume of the drug-loaded micelles; M is the amount of carrier input in the drug-loaded micelles

[0083] The determination of the aggregation number was based on the static light scattering method

[0084] A static light scattering (SLS) instrument combined with a differential detector was used to measure the molar mass Mw of the micelles. After toluene calibration, 5 samples of different concentrations were injected into the detector in order from low to high, with each injection amount being 2 ml, the flow rate being 0.5 ml / min, and the detection temperature being 25°C. After data collection, the data were analyzed by ASTRA software, and then the aggregation number was calculated according to the following formula.

[0085] Aggregation number = Mw / 2805

[0086] Residual solvent detection refers to the pharmacopoeia method

[0087] Example 1 Preparation and stability investigation of docetaxel nanomicelles

[0088] The prescription is shown in Table 1:

[0089] Table 1 Prescription of the docetaxel nanomicelle preparation of Example 1

[0090] Drug Lipid / drug (mass ratio) Drug concentration (mg / mL) Hydrating solvent Docetaxel 15:1 6 Deionized water Docetaxel 5:1 6 Deionized water

[0091] Preparation process: according to the above prescription, docetaxel and polyethylene glycol 2000 distearoyl phosphatidyl ethanolamine (PEG2000-DSPE, purchased from Avanti Company) were weighed in a rotary flask, and an appropriate amount of ethanol was added to gently shake the medicine and lipids to completely dissolve, to obtain a clear and transparent solution. The organic solvent in the rotary flask was dried on a vacuum rotary evaporator, the rotation speed was 90 r / min, and the water bath temperature was 40°C. After the organic solvent was dried, the lipids and drugs were uniformly distributed on the inner wall of the rotary flask. Then, after the rotary flask was cooled to room temperature, deionized water was added, and ultrasonic was performed in a 30°C water bath for 5 min (40 KHz, 50 w; same below), so that the lipid film was completely dissolved (drug concentration 6 mg / mL). An appropriate amount of freeze-drying excipient glucose was added to make the final concentration of glucose 125 mg / mL. The glucose was completely dissolved by gentle shaking, and then filtered through a 220 nm microporous filter, and then freeze-dried to obtain a docetaxel nanomicelle preparation freeze-dried powder with high drug loading, high encapsulation efficiency and high stability.

[0092] The molar mass of the polyethylene glycol 2000 distearoyl phosphatidyl ethanolamine empty micelle is about 2.52 x 10 5 gmol -1 , and the aggregation number is about 90. After loading the drug, the molar mass of the micelle increases with the increase of the drug loading, but the aggregation number does not change significantly, still about 90.

[0093] The obtained sample is a clear and transparent solution after reconstitution, the particle size distribution is between 5-20 nm, the average particle size is 12 nm, and the PDI is 0.107; the drug loading is 6.25% and 16.67% (mass percentage), respectively, the encapsulation efficiency is 98%-100%, and the storage stability (encapsulation efficiency 98%-100%) at 25°C is greater than 4 hours (of which 15:1, greater than 24 hours). (The aggregation number does not change significantly, still about 90).

[0094] Example 2: Preparation and stability investigation of cabazitaxel nanomicelle preparation

[0095] The prescription is shown in Table 2:

[0096] Table 2 Prescription of the cabazitaxel nanomicelle preparation of Example 2

[0097] Drug Lipid / drug (mass ratio) Drug concentration (mg / mL) Hydrating solvent Carboplatin 10:1 8 Deionized water

[0098] Preparation process: according to the above prescription proportion, cabazitaxel and polyethylene glycol 2000 distearoyl phosphatidyl ethanolamine were placed in a rotary flask, and an appropriate amount of ethanol was added to gently shake the drug and lipids to dissolve, to obtain a clear transparent solution, the organic solvent in the rotary flask was dried on a vacuum rotary evaporator, the rotation speed was 90 r / min, the water bath temperature was 40℃, after the organic solvent was dried, the lipids and drugs were evenly distributed on the inner wall of the rotary flask, then after the rotary flask was cooled to room temperature, deionized water was added, ultrasonic was performed at 30℃ water bath for 20 min, the lipid film was completely dissolved, an appropriate amount of freeze-drying excipient glucose was added, the final concentration of glucose was 125 mg / mL, gently shake to make the glucose completely dissolved, then filtered by 220 nm microporous filter membrane, freeze-dried, and the obtained sample was a clear transparent solution after reconstitution, the particle size distribution was between 5-20 nm, the average particle size was 12 nm, PDI was 0.114; the drug loading was 9.1%, the encapsulation efficiency was 98%-100%, and the storage stability (encapsulation efficiency 98%-100%) at 25℃ was more than 24 hours. (The aggregation number did not change obviously, still about 90)

[0099] Example 3: paclitaxel nanomicelles have high encapsulation efficiency, but poor stability and cannot meet the needs of clinical drug use

[0100] The prescription is shown in Table 3:

[0101] Table 3: Paclitaxel nanomicelle preparation prescription

[0102] Drug Lipid / drug (mass ratio) Drug concentration (mg / mL) Hydrating solvent Paclitaxel 15:1 5 Deionized water

[0103] Preparation process: according to the above prescription proportion, paclitaxel and polyethylene glycol 2000 distearoyl phosphatidyl ethanolamine were placed in a rotary flask, and an appropriate amount of ethanol was added to gently shake the drug and lipids to dissolve, to obtain a clear transparent solution, the organic solvent in the rotary flask was dried on a vacuum rotary evaporator, the rotation speed was 90 r / min, the water bath temperature was 40℃, after the organic solvent was dried, the lipids and drugs were evenly distributed on the inner wall of the rotary flask, then after the rotary flask was cooled to room temperature, deionized water was added, ultrasonic was performed at 30℃ water bath for 20 min, the lipid film was completely dissolved, then filtered by 220 nm microporous filter membrane (or, an appropriate amount of freeze-drying excipient glucose was added, the final concentration of glucose was 125 mg / ml, gently shake to make the glucose completely dissolved, and then filtered by 220 nm microporous filter membrane), the encapsulation efficiency was 99%, and then it was placed at room temperature to investigate its stability. It was found that paclitaxel nanomicelles had a tendency to become turbid after 10 min, the turbidity increased after 20 min, and a precipitate was precipitated, indicating that paclitaxel micelles were very unstable at room temperature and could not meet the needs of clinical drug use.

[0104] Example 4: Effect of lyophilization excipient on the reconstitution time and room temperature stability of Cabazitaxel nanomicelle lyophilized powder

[0105] The prescription is shown in Table 4:

[0106] Table 4 Cabazitaxel nanomicelle formulation prescription of Example 4

[0107] Drug Lipid / drug (mass ratio) Drug concentration (mg / mL) Hydrating solvent Carboplatin 10:1 8 Deionized water

[0108] Preparation process: according to the above prescription, the proportion of Cabazitaxel and polyethylene glycol 2000 distearoyl phosphatidyl ethanolamine was weighed and placed in a rotary evaporation flask, and an appropriate amount of ethanol was added to gently shake the drug and lipid to dissolve, to obtain a clear and transparent solution. The organic solvent in the rotary evaporation flask was dried on a vacuum rotary evaporator, the rotation speed was 90 r / min, and the water bath temperature was 40℃. After the organic solvent was dried, the lipid and drug were evenly distributed on the inner wall of the rotary evaporation flask. Then, after the rotary evaporation flask was cooled to room temperature, deionized water was added, and ultrasonic was performed at 30℃ water bath for 20 min to completely dissolve the lipid film. An appropriate amount of lyophilization excipient (glucose, mannitol, lactose or trehalose) was added to make the final concentration of lyophilization excipient 125 mg / mL. Gently shake to completely dissolve the lyophilization excipient, then filter with a 220 nm microporous filter, and freeze-dry. Take one bottle of nanomicelle lyophilized powder containing different lyophilization excipients, reconstitute with water for injection, and record the time required for Cabazitaxel micelle preparation to reconstitute to clear, as well as the clarity and encapsulation efficiency after being placed at room temperature (25℃) for 24 hours. The results are shown in Table 5. The test results show that the addition of lyophilization excipient does not affect the stability of the preparation, but the appropriate lyophilization excipient can significantly reduce the time required for the reconstitution of the lyophilized powder to completely dissolve. Considering the convenience of clinical use, we choose glucose as the lyophilization excipient of taxane micelle preparation!

[0109] Table 5 Reconstitution time and stability of Cabazitaxel nanomicelle preparation containing different lyophilization excipients

[0110]

[0111]

[0112] Stability greater than 100% is a normal error range due to measurement error.

[0113] Example 5 Comparison of 37℃ stability of Cabazitaxel nanomicelle preparation and Cabazitaxel injection (marketed)

[0114] The cabazitaxel nanomicelle preparation in Example 2 and the cabazitaxel injection were reconstituted or diluted with water for injection to a final drug concentration of 5 mg / mL, and then the two preparations were placed in a 37°C thermostat. The clarity of the preparations was observed at different time points. After 4 hours, drug crystallization was observed in the cabazitaxel injection (commercially available) solution, and by 8 hours, the drug crystallization increased and a large amount of precipitate appeared at the bottom, while the cabazitaxel nanomicelle preparation remained a clear and transparent liquid. At this time, the encapsulation efficiency was 99%, indicating that the stability of the cabazitaxel nanomicelle preparation was much better than that of the cabazitaxel injection (commercially available).

[0115] Example 6 Effect of the ratio (weight ratio) of lipids to drug on the stability of the taxane nanomicelle preparation

[0116] The prescription is shown in Table 6:

[0117] Table 6 Prescription of the cabazitaxel nanomicelle preparation of Example 6

[0118] Drug Lipid / drug (mass ratio) Drug concentration (mg / mL) Hydrating solvent Carboplatin 5:1 5 Deionized water Carboplatin 10:1 5 Deionized water Carboplatin 15:1 5 Deionized water

[0119] Preparation process: The cabazitaxel and the distearoylphosphatidyl ethanolamine of polyethylene glycol 2000 were weighed according to the above prescription ratio, placed in a rotary evaporation flask, and a suitable amount of ethanol was added to gently shake to dissolve the drug and lipids, obtaining a clear and transparent solution. The organic solvent in the rotary evaporation flask was dried on a vacuum rotary evaporator at a rotation speed of 90 r / min and a water bath temperature of 40°C. After the organic solvent was dried, the lipids and the drug were uniformly distributed on the inner wall of the rotary evaporation flask. Then, after the rotary evaporation flask was cooled to room temperature, deionized water was added, and ultrasonic treatment was performed in a 30°C water bath for 20 min to completely dissolve the lipid film. Then, a 220 nm microporous filter was used for filtration. Then, the cabazitaxel nanomicelle preparations with different drug-lipid ratios were placed at room temperature (25°C) to observe the change in clarity of the preparations and the stability during the observation period. The results are shown in Table 7.

[0120] Table 7 Effect of the ratio of lipids to drug on the stability of the cabazitaxel nanomicelle preparation

[0121]

[0122] When the same dose of drug is administered to a patient, the lower the drug loading of the nanomicelle preparation, the more carriers in the patient's body, and the greater the potential toxic side effects.

[0123] When 5:1, which is in a lyophilized form, is reconstituted with water for injection, and placed at 25°C for 2 hours, the clarity is a clear and transparent solution, the encapsulation efficiency (%) is 99.1%, and the particle size (nm) is 12.1, and the PDI is 0.107. Therefore, it can be configured into the required solution form 2 hours before use.

[0124] Example 7 Effect of different hydration temperatures on the room temperature stability of the cabazitaxel nanomicelle preparation

[0125] The prescription is shown in Table 8:

[0126] Table 8 Prescription of Cabazitaxel Nanomicelle Formulation in Example 7

[0127]

[0128] Preparation process: Cabazitaxel and polyethylene glycol 2000 distearoyl phosphatidyl ethanolamine were weighed according to the above prescription ratio and placed in a rotary evaporation flask. Appropriate amount of ethanol was added and gently shaken to dissolve the drug and lipids to obtain a clear and transparent solution. The organic solvent in the rotary evaporation flask was dried on a vacuum rotary evaporator at a speed of 90 r / min and a water bath temperature of 40℃. After the organic solvent was dried, the lipids and drugs were uniformly distributed on the inner wall of the rotary evaporation flask. Then, after the rotary evaporation flask was cooled to room temperature, deionized water was added and placed in a water bath at different temperatures for ultrasonic treatment for 20 min to completely dissolve the lipid film. After filtration with a 220 nm microporous filter, the cabazitaxel nanomicelle formulations prepared at different hydration temperatures were placed at room temperature (25℃) to observe the change in clarity of the formulation and the stability during the observation period. The results are shown in Table 9.

[0129] Table 9 Effect of Hydration Temperature on the Stability of Cabazitaxel Nanomicelle Formulation at Room Temperature

[0130]

[0131]

[0132] NA means not measured

[0133] Example 8: Other types of polymeric materials and taxane drugs form polymeric micelles with poor stability, which cannot meet the needs of clinical drug use

[0134] The prescription is shown in Table 10:

[0135] Table 10 Prescription of Other Types of Polymeric Nanomicelle Formulation in Example 8

[0136]

[0137] Process flow: according to the above prescription, cabazitaxel (CTX) and polymer material were weighed and placed in a rotary flask, and then an appropriate amount of chloroform was added and gently shaken to dissolve the CTX and polymer material, to obtain a clear and transparent solution. The organic solvent in the rotary flask was dried on a vacuum rotary evaporator, at a speed of 90 r / min and a water bath temperature of 40°C. After the organic solvent was dried, the polymer material and CTX were evenly distributed on the inner wall of the rotary flask. Then, after the rotary flask was cooled to room temperature, 10 mL of deionized water was added, and then ultrasonic was performed at 30°C to completely dissolve the lipid film. Then it was placed at room temperature (25°C) to investigate its stability, and the results are shown in Table 11. The test results suggest that although other types of polymer materials can also load taxane drugs, they generally have poor stability and cannot meet the requirements of clinical use, so they also have no application value.

[0138] Table 11 Stability of polymer micelles formed by other types of polymer materials and taxane drugs

[0139]

[0140]

[0141] PLGA is poly(lactic-co-glycolic acid), PDLLA is racemic polylactic acid, PCL is polycaprolactone, and F127 is poloxamer.

[0142] Example 9

[0143] Effect of cabazitaxel micelles on animal body weight

[0144] BALB / c mice (SPF level) were taken, 6-8 weeks old, weighing 19-21 g, and randomly divided into seven groups according to body weight, 10 in each group. The seven groups of mice were treated with cabazitaxel micelles (M-CTX) (Example 2) or cabazitaxel injection (CTX-Tween80) at the following concentrations: 12.5, 25 and 50 mg / kg of cabazitaxel micelles or cabazitaxel injection, tail vein administration once, and the effect of the drug on the body weight of the mice was recorded. The results are shown in Figure 1 According to the body weight results of the mice after administration, the toxicity of the cabazitaxel micelle preparation at the same dose was significantly lower than that of the cabazitaxel injection.

[0145] Table 12 Effect of different cabazitaxel preparations on mouse body weight

[0146]

[0147] Example 10

[0148] In vitro cell test

[0149] MTT (dimethylthiazole blue) staining method was used to determine the cytotoxicity of cabazitaxel micelles (Example 2) and cabazitaxel injection (self-made) (CTX-Tween80) on in vitro cultured MC-38. The in vitro cultured MC-38 cells (RPMI 1640 medium containing 10% fetal bovine serum, penicillin and streptomycin double antibiotic, culture environment 37°C, saturated humidity, carbon dioxide concentration 5%) were washed twice with PBS, trypsinized, counted with a cell counting plate, diluted to 2x10 4 cells per milliliter, and inoculated into a 96-well cell culture plate with 100 μL (2x10 3 cells) per well; incubated overnight in normal medium; 100 μL of complete medium containing different concentrations of cabazitaxel micelles and cabazitaxel injection was added to each well, with six parallel samples for each concentration, and incubated for 72 hours; the drug-containing medium was removed by PBS washing, 100 μL of MTT solution was added to each well, and incubated at 37°C for 4 hours; the MTT was discarded, 100 μL of dimethyl sulfoxide (DMSO) was added to each well, and the purple crystals were completely dissolved by gently shaking at room temperature for 10 minutes; the solution absorbance value was detected at 570 nm wavelength by a full-automatic enzyme label instrument (Thermo), and the cell growth curve under different drug concentrations was drawn. Figure 2

[0150] Table 13 Cytotoxic effect of cabazitaxel micelles on MC-38 cells

[0151]

[0152] The experimental results show that cabazitaxel micelles (M-CTX) have stronger growth inhibition effect on MC-38 cells than cabazitaxel injection (CTX-Tween80) at the same concentration, indicating that cabazitaxel micelles (M-CTX) have stronger killing effect on tumor cells than cabazitaxel injection (CTX-Tween80).

[0153] Example 11: Effect of lyophilization excipient on reconstitution time and room temperature storage stability of docetaxel nanomicelles lyophilized powder

[0154] The prescription is shown in the table:

[0155] Docetaxel nanomicelles formulation prescription

[0156]

[0157]

[0158] ​Preparation process: according to the above prescription proportion, docetaxel and polyethylene glycol 2000 distearoyl phosphatidyl ethanolamine are weighed in a rotary flask, and a proper amount of ethanol is added to gently shake to dissolve the drug and lipids to obtain a clear and transparent solution. The organic solvent in the rotary flask is dried on a vacuum rotary evaporator, the rotation speed is 90 r / min, and the water bath temperature is 40°C. After the organic solvent is dried, the lipids and drugs are uniformly distributed on the inner wall of the rotary flask. Then, after the rotary flask is cooled to room temperature, deionized water is added, and ultrasonic is performed at 30°C water bath for 20 min to completely dissolve the lipid film. A proper amount of freeze-drying excipient (glucose, mannitol, lactose or trehalose) is added to make the final concentration of freeze-drying excipient 125 mg / mL. Gently shake to completely dissolve the freeze-drying excipient, then filter with a 220 nm microporous filter, and freeze-dry. Take one bottle of each of the nanomicelle preparation freeze-dried powder containing different freeze-drying excipients, reconstitute with water for injection, record the time required for the docetaxel micelle preparation to reconstitute to be clear, and the clarity and encapsulation efficiency after being placed at room temperature (25°C) for 6 hours. The results are shown in the table. The test results show that the addition of freeze-drying excipients does not affect the stability of the preparation, but the appropriate freeze-drying excipient can significantly reduce the time required for the freeze-dried powder to reconstitute to completely dissolve.

[0159] Reconstitution time and stability of docetaxel nanomicelle preparation containing different freeze-drying excipients

[0160]

[0161] The stability of more than 100% is caused by measurement error, which is within the normal error range.

[0162] Example 12: Pharmacodynamic study of docetaxel nanomicelles on melanoma tumor model

[0163] Pharmacodynamic test of prepared docetaxel nanomicelles (Example 1) on human melanoma B16F10 C57 mouse subcutaneous transplanted tumor model: 20 qualified B16F10 C57 tumor animals were selected and randomly divided into 2 groups, 10 in each group, and given normal docetaxel injection (10 mg / kg) and docetaxel nanomicelles (10 mg / kg) respectively, tail vein injection, once every three days, a total of three times. Observe the general clinical symptoms of animals twice a day, and measure the tumor volume every 3 days (when the tumor volume exceeds 5000 mm 3 .

[0164] Time (days) / tumor volume (mm 3 )]]> DCX-TW80 M-DCX 1 268.8 239.1 4 745.2 368.7 7 1645.4 627.6 10 3132.8 1115.1 13 - 1494.7 16 - 2623.8 19 - 4603.3

[0165] "-" indicates that the tumor volume exceeds 5000 mm 3 .

[0166] The results of the drug efficacy are shown in the table. According to the results, the docetaxel nanomicelles have a significantly better effect on inhibiting tumor growth than the docetaxel injection.

[0167] Example 13: Cabazitaxel nanomicelles prepared by different types of PEG-PE

[0168] The prescription is shown in the table:

[0169] Prescription of cabazitaxel nanomicelles prepared by different types of PEG-PE

[0170]

[0171]

[0172] Preparation process: according to the above prescription, cabazitaxel and different types of PEG-PE were weighed in a rotary flask, and then an appropriate amount of ethanol was added to gently shake the drug and lipid to dissolve, to obtain a clear and transparent solution. The organic solvent in the rotary flask was dried on a vacuum rotary evaporator, the rotation speed was 90 r / min, and the water bath temperature was 40℃. After the organic solvent was dried, the lipid and drug were uniformly distributed on the inner wall of the rotary flask. Then, after the rotary flask was cooled to room temperature, deionized water was added, and ultrasonic was performed in a 25℃ water bath for 20 min to completely dissolve the lipid film. Then, a 220 nm microporous filter was used for filtration. Then, the cabazitaxel nanomicelles prepared by different drug-lipid ratios were placed at room temperature (25℃), and the change in clarity of the preparation was observed to investigate the stability, and the results are shown in the table.

[0173] Encapsulation efficiency and stability of cabazitaxel nanomicelles prepared by different types of PEG-PE

[0174]

Claims

1. A micelle comprising a taxane drug, characterized in that, 1 part of a taxane drug, 10-15 parts of polyethylene glycol derivatized phospholipid; the taxane drug is cabazitaxel; The polyethylene glycol derivatized phospholipid is polyethylene glycol 1000 distearoyl phosphatidyl ethanolamine, polyethylene glycol 2000 distearoyl phosphatidyl ethanolamine, polyethylene glycol 5000 distearoyl phosphatidyl ethanolamine, polyethylene glycol 2000 dioleoyl phosphatidyl ethanolamine, polyethylene glycol 2000 dipalmitoyl phosphatidyl ethanolamine or polyethylene glycol 2000 dimyristoyl phosphatidyl ethanolamine.

2. The micelle of claim 1, wherein, The micelle is a spherical micelle with a core-shell structure, wherein the inner core is a phospholipid layer and the outer shell is a polyethylene glycol layer, and the cabazitaxel is distributed in the polyethylene glycol derivatized phospholipid micelle; Or, the particle size of the micelle is 5-50 nm; Or, the polydispersity of the micelle is less than 0.

3.

3. The micelle of claim 2, wherein, 1 part of cabazitaxel and 10 or 15 parts of polyethylene glycol derivatized phospholipid; Or, the particle size of the micelle is 10-20 nm; Or, the polydispersity of the micelle is less than 0.

2.

4. The micelle of claim 1, wherein, The polyethylene glycol derivatized phospholipid is polyethylene glycol 2000 distearoyl phosphatidyl ethanolamine, and the average diameter of the micelle is 10-15 nm; Or, the polyethylene glycol derivatized phospholipid is polyethylene glycol 2000 distearoyl phosphatidyl ethanolamine, and the aggregation number of the micelle is 90.

5. The micelle of claim 1, wherein, The polyethylene glycol derivatized phospholipid is polyethylene glycol 2000 distearoyl phosphatidyl ethanolamine.

6. The micelle of claim 1, wherein, 1 part of cabazitaxel and 10 or 15 parts of polyethylene glycol 2000 distearoyl phosphatidyl ethanolamine.

7. The micelle of claim 1, wherein 1 part of cabazitaxel and 10 parts of polyethylene glycol 1000 distearoyl phosphatidyl ethanolamine, polyethylene glycol 5000 distearoyl phosphatidyl ethanolamine, polyethylene glycol 2000 dioleoyl phosphatidyl ethanolamine, polyethylene glycol 2000 dipalmitoyl phosphatidyl ethanolamine or polyethylene glycol 2000 dimyristoyl phosphatidyl ethanolamine.

8. A method of preparing the micelle of claim 1, wherein, It comprises the following steps: Step 1: removing the organic solvent in the solution to obtain a polymeric lipid film containing a taxane drug; wherein the solution is composed of 1 part of a taxane drug, 10-15 parts of polyethylene glycol derivatized phospholipid and the organic solvent; the taxane drug is cabazitaxel; the polyethylene glycol derivatized phospholipid is polyethylene glycol 1000 distearoyl phosphatidyl ethanolamine, polyethylene glycol 2000 distearoyl phosphatidyl ethanolamine, polyethylene glycol 5000 distearoyl phosphatidyl ethanolamine, polyethylene glycol 2000 dioleoyl phosphatidyl ethanolamine, polyethylene glycol 2000 dipalmitoyl phosphatidyl ethanolamine or polyethylene glycol 2000 dimyristoyl phosphatidyl ethanolamine; Step 2: hydrating the polymer lipid film obtained in step 1 in an aqueous solvent to obtain the micelles containing the taxane drug.

9. The preparation method of claim 8, wherein, the organic solvent is one or a mixture of at least two of an alcohol solvent, a halogenated hydrocarbon solvent and a nitrile solvent; or, the operation of removing the organic solvent is removing the organic solvent by a rotary evaporation method under reduced pressure; or, the aqueous solvent is one or a mixture of at least two of deionized water, a PBS buffer solution, normal saline, a glucose injection and an amino acid injection; or, the mass / volume ratio of the taxane drug in the aqueous solvent is 0.5 mg / mL-15 mg / mL; or, the operation of hydrating is vortex shaking, shaking or ultrasonic hydration; or, step 2 further comprises a filtration step: after hydration, the micelles containing the taxane drug are obtained by extrusion filtration through a nanoscale filter membrane; or, after hydration, the micelles containing the taxane drug are obtained by extrusion filtration through a nanoscale filter membrane after adding a pharmaceutical excipient.

10. The preparation method of claim 9, wherein, the organic solvent is an alcohol solvent, and the alcohol solvent is methanol and / or ethanol; or, the organic solvent is a halogenated hydrocarbon solvent, and the halogenated hydrocarbon solvent is chloroform; or, the organic solvent is a nitrile solvent, and the nitrile solvent is acetonitrile; or, the operation of removing the organic solvent is removing the organic solvent by a rotary evaporation method under reduced pressure, and the rotary evaporation temperature is 30°C-60°C; or, the operation of removing the organic solvent is removing the organic solvent by a rotary evaporation method under reduced pressure, and the rotary evaporation speed is 10-500 r / min; or, the aqueous solvent is deionized water; or, the mass / volume ratio of the taxane drug in the aqueous solvent is 5 mg / mL-10 mg / mL; or, the operation of hydrating is vortex shaking, and the time is 1-3 hours; or, the operation of hydrating is ultrasonic hydration, and the time is 1-60 minutes; Or, the hydration operation is ultrasonic hydration, and the ultrasonic intensity is 0.1-15 w / cm 2 ; or, the operation of hydrating is ultrasonic hydration, and the ultrasonic frequency is 0.1-100 kHz; or, the hydration temperature is 20°C-40°C; or, the pharmaceutical excipient is a freeze-drying excipient; or, the nanoscale filter membrane is selected from filter membranes with a pore size of 10-300 nm; or, the nanoscale filter membrane is made of a mixed cellulose ester microporous filter membrane, a nylon filter membrane, a polytetrafluoroethylene filter membrane, a polyvinylidene fluoride membrane, a polyether sulfone filter membrane or a polypropylene filter membrane.

11. The production method according to claim 9, wherein The preparation method comprises the following steps: dissolving the taxane drug, the polyethylene glycol derivatized phospholipid and the organic solvent in an organic solvent, rotary evaporation, removing the organic solvent, forming a uniform polymer lipid film, adding an aqueous solvent, ultrasonic hydration in a 20°C-40°C water bath, nanoscale filter membrane filtration, and obtaining the micelles containing the taxane drug; or, after hydration, mixing with a pharmaceutical excipient, nanoscale filter membrane filtration, and obtaining the micelles containing the taxane drug.

12. The production method according to claim 9, wherein The preparation method further comprises a freeze-drying step.

13. The preparation method of claim 10, wherein, the organic solvent is ethanol; or, the rotary evaporation temperature is 40℃±5℃; or, the rotary speed of the rotary evaporation is 60-150r / min; or, the mass-volume ratio of the taxane drug in the aqueous solvent is 5-8mg / mL; or, the hydration operation is ultrasonic hydration for 10-30 minutes; or, the hydration temperature is 30-40℃; or, the freeze-drying excipient is one or more of glucose, mannitol, lactose, trehalose, sucrose, dextran, glycine, fructose and sorbitol; or, the mass ratio of the freeze-drying excipient to the taxane drug is 15.6 to 25:1; or, the nanometer filter is selected from a filter with a pore size of 220nm; or, the material of the nanometer filter is polyvinylidene fluoride membrane or polyether sulfone filter membrane.

14. The preparation method of claim 13, wherein, the freeze-drying excipient is glucose; or, the mass ratio of the freeze-drying excipient to the taxane drug is 15.6:1 or 20.8:1; or, the material of the nanometer filter is polyether sulfone filter membrane.

15. A pharmaceutical composition comprising the taxane-containing micelles of any one of claims 1-7.

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