CN105640882A - Cabazitaxel micelle drug loading system, preparation method and applications thereof - Google Patents
Cabazitaxel micelle drug loading system, preparation method and applications thereof Download PDFInfo
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Abstract
The invention provides a cabazitaxel micelle drug loading system and a preparation method thereof and also discloses an application of the system in tumor treatment. The drug loading system is composed of 85 to 99.9 wt% of amphiphilic segmented copolymer and 0.1 to 15 wt% of cabazitaxel; wherein the amphiphilic segmented copolymer comprises hydrophilic chain segments and hydrophobic chain segments; the hydrophilic chain segments are composed of polyethylene glycol with a molecular weight of 100 to 6000 or methoxyl polyethylene glycol with a molecular weight of 300 to 6000, the hydrophobic chain segments are composed of polylactide with a molecular weight of 200 to 6000; the hydrophobic chain segments are terminated by groups containing fluorenyl methyl carboxyl and/or amino acid containing a benzene ring structure; and the weight ratio of polyethylene glycol or methoxyl polyethylene glycol to polylactide is 0.1-30:1. The invention also provides an application of the drug loading system in the preparation of drugs for treating tumor.
Description
Technical field
The present invention provides a kind of Cabazitaxel micelle medicine carrying system and its preparation method, and the application of its treatment tumour aspect disease, belongs to pharmaceutical field.
Background technology
Cabazitaxel is the derivative of new taxol, the derivative of the semi-synthetic compound of the 10-deacetylation bar card fourth III extracted from the natural elite stand of yew of Europe, belong to third generation taxanes anti-tumor compounds, its mechanism of action and taxanes are seemingly, by strengthening tubulin polymerization effect and suppress microtubule depolymerization effect, cause forming stable non-functional microtubule fasolculus, thus destroy the mitotic division of tumour cell, the differentiation of inhibition tumor cell; Antiproliferative activity in Vitro Tumor model has the drug effect identical with docetaxel, the same with docetaxel effective facilitate ira vitro tube albumen polymerization and microtubule can be stablized in cold induction unzipping.
The molecular formula of Cabazitaxel: C45H57NO14, C3H6O molecular weight is 835.93 (without solvents), and structural formula is as follows:
Polymer micelle is a kind of stable colloidal dispersion system that development in recent years is got up. Micella is formed by self-assembly by parents' property block macromolecular polymkeric substance usually, and its hydrophobic segment is inside, and hydrophilic chain is outside, presents typical " core-shell structure copolymer " structure. Polymer micelle can significantly improve the solubleness of insoluble drug; Hydrophobic pharmaceutical pack is wrapped in the core of micella, it is possible to reduce the toxic side effects of medicine; In addition, micella passs release system as a kind of Nano medication, reaches the object of tumor tissues passive target by strengthening osmotic effect (EnhancedPermeabilityandRetentionEffect, EPR effect).
In order to effectively play the EPR effect of nano-carrier, it is to increase the tumor tissue targeting of medicine, just requiring that carrier can keep stable in blood, the circulation long-time in blood by nano-carrier improves the probability entering tumor tissues. But, for self-assembling polymers micella, research in recent years finds: in the very short time of micella after entering blood, medicine just discharges, and the medicine of release completes the transhipment (JournalofControlledRelease164 (2012): 108-114) of tissue and tumour essentially by the mode of random diffusion. The major cause this situation occur is that the hydrophobic segment in block macromolecular and the reactive force between medicine are more weak, and micella is through the dilution of blood after entering blood, and medicine is easy to from the core of micella to diffuse out. It thus is seen that to be improved the stability of micella, thus more effective performance passive target effect, the problem of core is the reactive force increased between hydrophobic segment and drug molecule.
For above-mentioned technical problem, contriver is by the collection of feasibility in theory demonstration and a large amount of test-results, provide a kind of Cabazitaxel micelle medicine carrying system, except the Cabazitaxel containing significant quantity, this system further comprises the chimeric multipolymer of a kind of novel amphiphilic, this multipolymer is made up of jointly hydrophilic section and hydrophobic section, hydrophilic section has selected polyoxyethylene glycol (PEG) or methoxy poly (ethylene glycol) (mPEG), these two kinds have polymkeric substance and have good biocompatibility and biodegradability, hydrophobic section then needs with polylactide, thus construct PEG-PLA or mPEG-PLA polymkeric substance, and on the terminal hydroxyl of PEG-PLA or mPEG-PLA, introduce fluorenylmethyloxycarbonyl (Fmoc) group, owing in Fmoc group, the delocalization scope of electronic cloud is very big, therefore, it is possible to and containing phenyl ring, very strong conjugation is formed between the medicine of the groups such as double bond, and Cabazitaxel is also exactly the medicine being rich in phenyl ring, therefore prepare high molecular polymer with this and can have very strong interaction with Cabazitaxel, and drug molecule is limited firmly " in core " the not easily stripping in micella, obtained a kind of inside and outside has the micelle medicine carrying system of high stability, the targeting of the tumor tissues of medicine can be improved.
Summary of the invention
According to specification sheets technical background content, technical problem solved by the invention is, it is to increase the reactive force between polymer micelle and Cabazitaxel, thus makes it in drug effect process, existence that can be highly stable, it is to increase Cabazitaxel is for the targeting of tumor tissues.
Based on above-mentioned purpose, the present invention provides a kind of Cabazitaxel micelle medicine carrying system, described drug-loading system is made up of the Cabazitaxel of massfraction to be the amphipathic nature block polymer of 85%-99.9% and massfraction be 0.1-15%, described amphipathic nature block polymer comprises hydrophilic segment and hydrophobic chain segment, hydrophilic segment is the polyoxyethylene glycol of molecular weight 100-6000 or the methoxy poly (ethylene glycol) of molecular weight 300-6000, hydrophobic chain is the polylactide of molecular weight 200-6000, hydrophobic chain carries out end-blocking with the group containing fluorenes first oxygen carboxyl and/or the amino acid containing benzene ring structure, the part by weight of polyoxyethylene glycol or methoxy poly (ethylene glycol) and polylactide is 0.1-30: 1.
In amphipathic chimeric micelle medicine carrying system, hydrophobic chain can also carry out end-blocking with other amino acid; Further preferably, hydrophobic chain can carry out end-blocking by phenylalanine.
Above-mentioned drug-loading system, wherein, the part by weight of polyoxyethylene glycol or methoxy poly (ethylene glycol) and polylactide is 0.5-10: 1.
Owing to methoxy poly (ethylene glycol) is different with the molecular weight of polylactide, cause their composition drug-loading system stability variant. So, above-mentioned drug-loading system, it may be preferred that hydrophilic chain is the methoxy poly (ethylene glycol) of molecular weight 300-4000, hydrophobic chain is the polylactide of molecular weight 300-5000; More preferably, hydrophilic chain is the methoxy poly (ethylene glycol) of molecular weight 600-3000, and hydrophobic chain is the polylactide of molecular weight 400-4000; Wherein, the part by weight of polyoxyethylene glycol or methoxy poly (ethylene glycol) and polylactide is 0.5-10: 1.
In a preferred embodiment of the invention, above-mentioned drug-loading system is made up of the Cabazitaxel of massfraction to be the amphipathic nature block polymer of 90%-97% and massfraction be 3-10%; Described hydrophilic segment is molecular weight is the methoxy poly (ethylene glycol) of 1500-3000, hydrophobic chain is the polylactide of molecular weight 1000-2000, this hydrophobic chain carries out end-blocking with the group containing fluorenes first oxygen carboxyl and/or the amino acid containing benzene ring structure, and the part by weight of methoxy poly (ethylene glycol) and polylactide is 0.5-10: 1.
Above-mentioned drug-loading system, wherein hydrophobic chain carries out end-blocking with the group containing fluorenes first oxygen carboxyl and/or the amino acid containing benzene ring structure, and the group of described fluorenes first oxygen carboxyl can be the arbitrary or combination of 6-fluorenylmethyloxycarbonyl amino or 6-fluorenylmethyloxycarbonyl amino oxygen carbonylamino caproic acid. Further preferably, hydrophobic chain carries out end-blocking with the arbitrary or combination of 6-fluorenylmethyloxycarbonyl amino-2-t-butoxycarbonyl amino caproic acid, 6-fluorenylmethyloxycarbonyl amino-2-benzyloxycarbonyl amino caproic acid, 6-fluorenylmethyloxycarbonyl amino-2-benzyloxycarbonyl amino phenylalanine, 6-fluorenylmethyloxycarbonyl amino-2-t-butoxycarbonyl amino phenylalanine.
Group containing fluorenes first oxygen carboxyl of the present invention is more preferably arbitrary or its combination in 6-fluorenylmethyloxycarbonyl amino, 6-fluorenylmethyloxycarbonyl amino-2-t-butoxycarbonyl amino caproic acid, 6-fluorenylmethyloxycarbonyl amino-2-benzyloxycarbonyl amino caproic acid.
In a preferred embodiment of the invention, above-mentioned drug-loading system is made up of the Cabazitaxel of massfraction to be the amphipathic nature block polymer of 85%-99.9% and massfraction be 0.1-15%, preferably, above-mentioned drug-loading system is made up of the Cabazitaxel of massfraction to be the amphipathic nature block polymer of 88%-98% and massfraction be 2-12%, namely, the drug loading 2%-12% of multipolymer, more preferably, it is made up of the Cabazitaxel of massfraction to be the amphipathic nature block polymer of 90%-97% and massfraction be 3-10%;Described hydrophilic segment is molecular weight is the methoxy poly (ethylene glycol) of 1500-3000, and hydrophobic chain is the polylactide of molecular weight 1000-2000, and this hydrophobic chain carries out end-blocking with the group containing fluorenes first oxygen carboxyl and/or the amino acid containing benzene ring structure.
Preferably, the described group containing fluorenes first oxygen carboxyl is 6-fluorenylmethyloxycarbonyl amino-2-benzyloxycarbonyl amino caproic acid and/or 6-fluorenylmethyloxycarbonyl amino-2-t-butoxycarbonyl amino caproic acid.
In general micelle medicine carrying system, the drug loading of multipolymer is no more than 5%, drug loading higher than 5% in the field of business comparison is difficult to accomplish, in the drug-loading system of the present invention, the drug loading of multipolymer can meet or exceed 5%, the highest more than 10%, improve the reactive force between polymer micelle and Cabazitaxel, thus make it can be highly stable in drug effect process existence, it is to increase Cabazitaxel is for the targeting of tumor tissues. This is the obvious useful effect of the present invention, is also one of innovative point of the present invention.
Above-mentioned drug-loading system can effectively solve micella hydrophobic chain and drug interaction power weak, the technical barrier of drug-loading system stability difference, this is the useful effect of the another aspect of the present invention.
In addition, now for clinical Cabazitaxel preparation, using Polysorbate 80 as solubility promoter, make thick injection concentrated solution, production technique is for adopting Polysorbate 80 and dissolve with ethanol, then freeze-drying removing ethanol, and technique is dangerous big, need before freeze-drying to ensure aseptic technique, not only operate loaded down with trivial details and cost is huge. In addition, research in recent years is it has been found that Polysorbate 80 easily causes anaphylaxis in body, and can cause the side effect such as hematotoxicity and fluid retention, and therefore before Clinical practice, patient need to carry out desensitization process and close observation infusion process.
Therefore required drug-loading system can adopt freeze-dried preparation administration just seem particularly important.
Cabazitaxel micelle medicine carrying system provided by the invention, except the Cabazitaxel containing significant quantity, further comprises the chimeric multipolymer of a kind of novel amphiphilic, this multipolymer is made up of jointly hydrophilic section and hydrophobic section, hydrophilic section has selected polyoxyethylene glycol (PEG) or methoxy poly (ethylene glycol) (mPEG), these two kinds have polymkeric substance and have good biocompatibility and biodegradability, hydrophobic section then needs with polylactide, thus construct PEG-PLA or mPEG-PLA polymkeric substance, and on the terminal hydroxyl of PEG-PLA or mPEG-PLA, introduce fluorenylmethyloxycarbonyl (Fmoc) group, owing in Fmoc group, the delocalization scope of electronic cloud is very big, therefore, it is possible to and containing phenyl ring, very strong conjugation is formed between the medicine of the groups such as double bond, and Cabazitaxel is also exactly the medicine being rich in phenyl ring, therefore prepare high molecular polymer with this and can have very strong interaction with Cabazitaxel, and drug molecule is limited firmly " in core " the not easily stripping in micella, obtained a kind of inside and outside has the micelle medicine carrying system of high stability, the targeting of the tumor tissues of medicine can be improved.
In an embodiment of the present invention, it provides the preparation method of a kind of drug-loading system, comprises
(1) polyoxyethylene glycol or methoxy poly (ethylene glycol) are joined in polymerization bottle, intensification is stirred and is vacuumized, rac-Lactide is added after cooling, stannous octoate, reactant high temperature polymerization after product methylene dichloride dissolves, and filters to obtain solid polyethylene glycol or methoxy poly (ethylene glycol) block polymer after precipitation;
(2) triethylamine is added after being dissolved by fluorenylmethyloxycarbonyl amino anhydrous tetrahydro furan, adding pivalyl chloride after cooling, have precipitation to generate, reactant continues reaction after heating up, after being filtered to remove impurity, evaporating solvent is to dry, obtains new penta acid anhydride that thick liquid is fluorenylmethyloxycarbonyl amino;
(3) polyoxyethylene glycol or methoxy poly (ethylene glycol) block polymer are dissolved in methylene dichloride, add triethylamine, 4-pyrollidinopyridine, after this solution is cooled, new penta acid anhydride of the fluorenylmethyloxycarbonyl amino of above-mentioned reaction gained is dissolved in methylene dichloride, after join in the dichloromethane solution of polyoxyethylene glycol or methoxy poly (ethylene glycol) block polymer, heat up after stirring, continue stirring reaction. Reactant precipitate after filter solid is the amino-terminated polyoxyethylene glycol of fluorenylmethyloxycarbonyl or methoxy poly (ethylene glycol)-polylactide block polymer;
(4) polymkeric substance and Cabazitaxel are made micellar solution.
In a preferred embodiment of the invention, additionally provide the preparation method of a kind of drug-loading system, comprising:
(1) synthesis of mPEG-PLA
Polyoxyethylene glycol or methoxy poly (ethylene glycol) are joined in polymerization bottle, intensification is stirred and is vacuumized, and adds rac-Lactide, stannous octoate after cooling, reactant dissolves through high temperature polymerization after product methylene dichloride, filters to obtain solid polyethylene glycol or methoxy poly (ethylene glycol) block polymer after precipitation;
(2) acid anhydrides preparation
Triethylamine is added after being dissolved by 6-fluorenylmethyloxycarbonyl amino-2-t-butoxycarbonyl amino caproic acid anhydrous tetrahydro furan, pivalyl chloride is added after cooling, precipitation is had to generate, reactant continues reaction after heating up, after being filtered to remove impurity, evaporating solvent is to dry, and obtaining thick liquid is 6-fluorenylmethyloxycarbonyl amino new penta acid anhydride of-2-t-butoxycarbonyl amino caproic acid;
(3) end capping
Polyoxyethylene glycol or methoxy poly (ethylene glycol) block polymer are dissolved in methylene dichloride, add triethylamine, 4-pyrollidinopyridine, after this solution is cooled, 6-fluorenylmethyloxycarbonyl amino new penta acid anhydride of-2-t-butoxycarbonyl amino caproic acid of above-mentioned reaction gained is dissolved in methylene dichloride, after join in the dichloromethane solution of polyoxyethylene glycol or methoxy poly (ethylene glycol) block polymer, heat up after stirring, continue stirring reaction. Reactant filter after precipitating solid is polyoxyethylene glycol or the methoxy poly (ethylene glycol)-polylactide block polymer of 6-fluorenylmethyloxycarbonyl amino-2-t-butoxycarbonyl amino caproic acid end-blocking;
(4) micelle freeze-drying synthesis
By polymkeric substance, Cabazitaxel and and suitable excipient substance make micellar solution after carry out freeze-drying.
According to above-mentioned preparation method, present invention also offers a kind of preferred method, comprise the following steps:
(1) 10g methoxy poly (ethylene glycol) 2000 joins in polymerization bottle, intensification is stirred and is vacuumized, and adds rac-Lactide 11g, stannous octoate 5.5mg after cooling, reactant high temperature polymerization after product methylene dichloride dissolves, filter after precipitation white solid both for mPEG-PLA block polymer;
(2) 11.7g6-fluorenylmethyloxycarbonyl amino-2-t-butoxycarbonyl amino caproic acid anhydrous tetrahydro furan adds 3-10mL triethylamine after dissolving, and adds pivalyl chloride 2-5mL after cooling, has precipitation to generate. Reactant continues reaction 1h after heating up, after being filtered to remove impurity, evaporating solvent is to dry, and obtaining thick liquid is 6-fluorenylmethyloxycarbonyl amino new penta acid anhydride of-2-t-butoxycarbonyl amino caproic acid;
(3) 20gmPEG-PLA is dissolved in methylene dichloride, add triethylamine 5mL, 4-pyrollidinopyridine 1.0g, after this solution is cooled, 6-fluorenylmethyloxycarbonyl amino new penta acid anhydride of-2-t-butoxycarbonyl amino caproic acid of above-mentioned reaction gained is dissolved in methylene dichloride, after join in the dichloromethane solution of mPEG-PLA, heat up after stirring reaction 1h, continue stirring reaction. Reactant filter after precipitating solid had both been the methoxy poly (ethylene glycol)-polylactide block polymer (mPEG-PLA-Lys (Fmoc)) of 6-fluorenylmethyloxycarbonyl amino-2-t-butoxycarbonyl amino caproic acid end-blocking.
Present invention also offers the preparation method that described drug-loading system is prepared into freeze-dried preparation, comprising:
By freeze-dried mixed to micellar solution obtained by above-mentioned steps (4) and freeze-drying auxiliary material, i.e. obtained freeze-dried preparation.
The step of aseptically process is also had before preferred described step of freeze drying.
Above-mentioned synthesis route is as follows:
(4) by polymkeric substance, Cabazitaxel and and suitable excipient substance make micellar solution after carry out freeze-drying.
According to method described above, wherein excipient substance is freeze-dried excipient, it is preferable that have aseptically process step before step of freeze drying. Described freeze-dried excipient can be one or more combinations of lactose, N.F,USP MANNITOL, sorbyl alcohol, Xylitol, sucrose, trehalose, fructose, glucose, sodium alginate or gelatin, it is preferable that lactose or N.F,USP MANNITOL.
According to above-mentioned preparation method, preferred 60mg Cabazitaxel and 1140mgmPEG-PLA-Lys (Fmoc) are dissolved in 20mL ethanol at 45 DEG C, rotary evaporation adds 20mL ultrapure water and dissolves medicine film after removing ethanol, after gained solution adds 400mg N.F,USP MANNITOL after 0.22um membrane filtration, freeze-drying obtains Cabazitaxel micelle freeze-drying powder.
The freeze-dried preparation of a kind of Cabazitaxel micelle medicine carrying system can have been prepared by aforesaid method.
Present invention also offers the freeze-dried preparation containing above-mentioned drug-loading system, described freeze-dried preparation is made up of drug-loading system and freeze-drying auxiliary material, Cabazitaxel and freeze-drying auxiliary material weight ratio be 1: 0.1-50, preferably, the weight ratio of Cabazitaxel and freeze-drying auxiliary material is 1: 0.5-30, more preferably, the weight ratio of Cabazitaxel and freeze-drying auxiliary material is 1: 1-10. Described freeze-drying auxiliary material is one or more combinations of lactose, N.F,USP MANNITOL, Xylitol, sorbyl alcohol, sucrose, trehalose, fructose, glucose, sodium alginate or gelatin.
In a preferred embodiment of the invention, in amphipathic nature block polymer described in above-mentioned drug-loading system, the weight ratio of Cabazitaxel and pharmaceutical excipients is 1: 0.5-30, described multipolymer drug loading is 2-12%, described hydrophilic segment is molecular weight is the methoxy poly (ethylene glycol) of 1500-3000, hydrophobic chain is the polylactide of molecular weight 1000-2000, and this hydrophobic chain carries out end-blocking with the group containing fluorenes first oxygen carboxyl; Described excipient substance is lactose or N.F,USP MANNITOL.
The present invention is loaded with the polymer micelle freeze-dried preparation of Cabazitaxel, substantially increase the solubleness of Cabazitaxel, and the micella particle diameter of preparation is little, encapsulation rate height, good stability, has the feature such as passive target and long circulating, wherein mPEG-PLA-Lys (Fmoc) block polymer has the effect overcoming multidrug resistance, the accumulation of the micella energy significantly increasing medicament being prepared in cell, can be applicable to reversing tumor many resistances field, has good application prospect.
Present invention also offers the application of the drug-loading system containing Cabazitaxel in the medicine of preparation treatment tumour.
Accompanying drawing explanation
Fig. 1 is the molecular weight and the molecular weight distribution coefficient figure that adopt GPC method to measure polymer.
Fig. 2 is the nucleus magnetic hydrogen spectrum figure of Fmoc.
Embodiment
Contriver verifies the practicality of the compounds of this invention and useful effect by following experiment.
Again reaffirm: following experiment is the exemplary experiment in numerous experiment in R&D process of the present invention, not contained with limit all experiments that invention people does for the present invention, object is only that setting forth the present invention by those data is provided with features and application.
Embodiment 1
The preparation of methoxy poly (ethylene glycol)-polylactide block copolymer
1)mPEG2000-PLA1300Synthesis
10g methoxy poly (ethylene glycol) 2000 joins in polymerization bottle, it is warming up to 130 DEG C to stir and vacuumize 1h, D is added after being cooled to room temperature, L-rac-Lactide 11g, stannous octoate 5.5mg, vacuum seal polymerization bottle, reactant is polymerized 15h after product methylene dichloride at 130 DEG C and dissolves, filter after ice ether sedimentation white solid both for mPEG2000-PLA1300Segmented copolymer, it is 3300 that core magnetic calculates polymericular weight, and gel permeation chromatography (GelPermeationChromatography, GPC) measures the molecular weight of polymer and molecular weight distribution coefficient is respectively 4596,1.06.(see Fig. 1 and data table related 1).
Table 1 broad sense relative peak
2) end capping
11.7g6-fluorenylmethyloxycarbonyl amino-2-t-butoxycarbonyl amino caproic acid 50mL anhydrous tetrahydro furan adds 3.5mL triethylamine after dissolving, and adds pivalyl chloride 3.05mL after being cooled to-10 DEG C, and adularescent precipitation generates at once. Reactant is warming up to and 0 DEG C of reaction 2h follow-up continuing is warming up to 25 DEG C of reaction 1h, and after being filtered to remove insolubles, evaporating solvent is to dry that colourless viscous liquid is 6-fluorenylmethyloxycarbonyl amino new penta acid anhydride of-2-t-butoxycarbonyl amino caproic acid.
20gmPEG2000-PLA1300It is dissolved in 100mL methylene dichloride, add triethylamine 3.5mL, 4-pyrollidinopyridine 0.37g, after this solution is cooled to 0 DEG C, join after 6-fluorenylmethyloxycarbonyl amino new penta acid anhydride of-2-t-butoxycarbonyl amino caproic acid of above-mentioned reaction gained is dissolved in 50mL methylene dichloride in the dichloromethane solution of mPEG-PLA, it is warming up to 25 DEG C after stirring reaction 1h and continues stirring reaction 24h. Filter after reactant ice ether sedimentation white solid had both been the methoxy poly (ethylene glycol)-polylactide block copolymer (mPEG of 6-fluorenylmethyloxycarbonyl amino-2-t-butoxycarbonyl amino caproic acid end-blocking2000-PLA1300-Lys (Fmoc)), nucleus magnetic hydrogen spectrum is shown in Fig. 2.
Embodiment 2
1)mPEG2000-PLA1000Synthesis
10g methoxy poly (ethylene glycol) 2000 joins in polymerization bottle, it is warming up to 130 DEG C to stir and vacuumize 1h, D is added after being cooled to room temperature, L-rac-Lactide 7g, stannous octoate 3mg, vacuum seal polymerization bottle, reactant is polymerized 15h after product methylene dichloride at 130 DEG C and dissolves, filter after ice ether sedimentation white solid both for mPEG2000-PLA1000Segmented copolymer, it is that 3000, GPC measures the molecular weight of polymer and molecular weight distribution coefficient is respectively 3943,1.05 that core magnetic calculates polymericular weight.
2) end capping
11.7g6-fluorenylmethyloxycarbonyl amino-2-t-butoxycarbonyl amino caproic acid 50mL anhydrous tetrahydro furan adds 3-10mL triethylamine after dissolving, and adds pivalyl chloride 2-5mL after being cooled to-10 DEG C, and adularescent precipitation generates at once. Reactant is warming up to and 0 DEG C of reaction 2h follow-up continuing is warming up to 25 DEG C of reaction 1h, and after being filtered to remove insolubles, evaporating solvent is to dry that colourless viscous liquid is 6-fluorenylmethyloxycarbonyl amino new penta acid anhydride of-2-t-butoxycarbonyl amino caproic acid.
20gmPEG2000-PLA1000It is dissolved in 100mL methylene dichloride, add triethylamine 3.5mL, 4-pyrollidinopyridine 0.37g, after this solution is cooled to 0 DEG C, join after 6-fluorenylmethyloxycarbonyl amino new penta acid anhydride of-2-t-butoxycarbonyl amino caproic acid of above-mentioned reaction gained is dissolved in 50mL methylene dichloride in the dichloromethane solution of mPEG-PLA, it is warming up to 25 DEG C after stirring reaction 1h and continues stirring reaction 24h. Filter after reactant ice ether sedimentation white solid had both been the methoxy poly (ethylene glycol)-polylactide block copolymer (mPEG of 6-fluorenylmethyloxycarbonyl amino-2-t-butoxycarbonyl amino caproic acid end-blocking2000-PLA1000-Lys(Fmoc))
Embodiment 3
1)mPEG2000-PLA1500Synthesis
10g methoxy poly (ethylene glycol) 2000 joins in polymerization bottle, it is warming up to 130 DEG C to stir and vacuumize 1h, D is added after being cooled to room temperature, L-rac-Lactide 13g, stannous octoate 6mg, vacuum seal polymerization bottle, reactant is polymerized 15h after product methylene dichloride at 130 DEG C and dissolves, filter after ice ether sedimentation white solid both for mPEG2000-PLA1300Segmented copolymer, it is that 3500, GPC measures the molecular weight of polymer and molecular weight distribution coefficient is respectively 4746,1.06 that core magnetic calculates polymericular weight.
2) end capping
11.7g6-fluorenylmethyloxycarbonyl amino-2-t-butoxycarbonyl amino caproic acid 50mL anhydrous tetrahydro furan adds 3.5mL triethylamine after dissolving, and adds pivalyl chloride 3.05mL after being cooled to-10 DEG C), adularescent precipitation generates at once.Reactant is warming up to and 0 DEG C of reaction 2h follow-up continuing is warming up to 25 DEG C of reaction 1h, and after being filtered to remove insolubles, evaporating solvent is to dry that colourless viscous liquid is 6-fluorenylmethyloxycarbonyl amino new penta acid anhydride of-2-t-butoxycarbonyl amino caproic acid.
20gmPEG2000-PLA1500It is dissolved in 100mL methylene dichloride, add triethylamine 3.5mL, 4-pyrollidinopyridine 0.37g, after this solution is cooled to 0 DEG C, join after 6-fluorenylmethyloxycarbonyl amino new penta acid anhydride of-2-t-butoxycarbonyl amino caproic acid of above-mentioned reaction gained is dissolved in 50mL methylene dichloride in the dichloromethane solution of mPEG-PLA, it is warming up to 25 DEG C after stirring reaction 1h and continues stirring reaction 24h. Filter after reactant ice ether sedimentation white solid had both been the methoxy poly (ethylene glycol)-polylactide block copolymer (mPEG of 6-fluorenylmethyloxycarbonyl amino-2-t-butoxycarbonyl amino caproic acid end-blocking2000-PLA1500-Lys(Fmoc))
Embodiment 4
1)mPEG2000-PLA1800Synthesis
10g methoxy poly (ethylene glycol) 2000 joins in polymerization bottle, it is warming up to 130 DEG C to stir and vacuumize 1h, D is added, L-rac-Lactide 15g, stannous octoate 8mg after being cooled to room temperature, vacuum seal polymerization bottle, reactant is polymerized 15h after product methylene dichloride at 130 DEG C and dissolves, filter after ice ether sedimentation white solid both for mPEG2000-PLA1800 segmented copolymer, it is 3800 that core magnetic calculates polymericular weight, molecular weight and the molecular weight distribution coefficient of GPC mensuration polymer are respectively 4938,1.06.
2) end capping
11.7g6-fluorenylmethyloxycarbonyl amino-2-t-butoxycarbonyl amino caproic acid 50mL anhydrous tetrahydro furan adds 3.5mL triethylamine after dissolving, and adds pivalyl chloride 3.05mL after being cooled to-10 DEG C, and adularescent precipitation generates at once. Reactant is warming up to and 0 DEG C of reaction 2h follow-up continuing is warming up to 25 DEG C of reaction 1h, and after being filtered to remove insolubles, evaporating solvent is to dry that colourless viscous liquid is 6-fluorenylmethyloxycarbonyl amino new penta acid anhydride of-2-t-butoxycarbonyl amino caproic acid.
20gmPEG2000-PLA1800It is dissolved in 100mL methylene dichloride, add triethylamine 3.5mL, 4-pyrollidinopyridine 0.37g, after this solution is cooled to 0 DEG C, join after 6-fluorenylmethyloxycarbonyl amino new penta acid anhydride of-2-t-butoxycarbonyl amino caproic acid of above-mentioned reaction gained is dissolved in 50mL methylene dichloride in the dichloromethane solution of mPEG-PLA, it is warming up to 25 DEG C after stirring reaction 1h and continues stirring reaction 24h. Filter after reactant ice ether sedimentation white solid had both been the methoxy poly (ethylene glycol)-polylactide block copolymer (mPEG of 6-fluorenylmethyloxycarbonyl amino-2-t-butoxycarbonyl amino caproic acid end-blocking2000-PLA1800-Lys(Fmoc))��
Embodiment 5
20mg Cabazitaxel and the mPEG obtained by embodiment 12000-PLA1300-Lys (Fmoc) 80mg is dissolved in 20mL ethanol at 45 DEG C, rotary evaporation adds 2mL ultrapure water and dissolves medicine film after removing ethanol, after gained solution adds 500mg N.F,USP MANNITOL after 0.22um membrane filtration, freeze-drying obtains Cabazitaxel micelle freeze-drying powder, drug loading 2%.
Embodiment 6
60mg Cabazitaxel and the obtained mPEG of embodiment 12000-PLA1300-Lys (Fmoc) 1140mg is dissolved in 20mL ethanol in 45 DEG C, rotary evaporation dissolves medicine film except adding 20mL ultrapure water after ethanol, after gained solution adds 300mg N.F,USP MANNITOL after 0.22um membrane filtration, freeze-drying obtains Cabazitaxel micelle freeze-drying powder, drug loading 5%.
Embodiment 7
80mg Cabazitaxel and the mPEG obtained by embodiment 22000-PLA1000-Lys (Fmoc) 455mg is dissolved in 20mL ethanol at 45 DEG C, rotary evaporation adds 2mL ultrapure water and dissolves medicine film after removing ethanol, after adding 10mg N.F,USP MANNITOL, the freeze-drying after 0.22um membrane filtration of gained solution obtains Cabazitaxel micelle freeze-drying powder, drug loading 15%.
Embodiment 8
30mg Cabazitaxel and the mPEG obtained by embodiment 32000-PLA1500-Lys (Fmoc) 270mg is dissolved in 5mL ethanol at 45 DEG C, rotary evaporation adds 10mL ultrapure water and dissolves medicine film after removing ethanol, after gained solution adds 300mg sucrose after 0.22um membrane filtration, freeze-drying obtains Cabazitaxel micelle freeze-drying powder, drug loading 10%.
Embodiment 9
10mg Cabazitaxel and the mPEG obtained by embodiment 42000-PLA1800-Lys (Fmoc) 990mg is dissolved in 5mL ethanol at 45 DEG C, rotary evaporation adds 5mL ultrapure water and dissolves medicine film after removing ethanol, after gained solution adds 10mg glucose after 0.22um membrane filtration, freeze-drying obtains Cabazitaxel micelle freeze-drying powder, drug loading 1%.
Embodiment 10
Prepare mPEG respectively2000-PLA1300/ Cabazitaxel micellar solution and mPEG2000-PLA1300-Lys (Fmoc)/Cabazitaxel micellar solution (drug level is 2mg/mL), in two kinds of micellar solution, add foetal calf serum (FBS) respectively and regulate serum-concentration to 50%, dynamic light scattering is adopted to measure the change of size of different time micella, the initial particle of two kinds of Cabazitaxel micellas all at about 21nm, then mPEG2000-PLA1300-Lys (Fmoc)/Cabazitaxel micella in the blood solution of 50% after 12h particle diameter still there is no considerable change; But mPEG2000-PLA1300The micella size distribution that/Cabazitaxel micella records in the serum solution of 50% obviously becomes wide, illustrates that micella there occurs gathering. It thus is seen that mPEG2000-PLA1300-Lys (Fmoc)/Cabazitaxel micella has very high stability.
Ultrapure water described in the present invention is water for injection, is known to the skilled person.
The present invention is not limited to above-mentioned enforcement mode, anyone other any products identical or close with the present invention drawn under the enlightenment of the present invention, is all not precluded within outside protection scope of the present invention.
Claims (14)
1. a Cabazitaxel micelle medicine carrying system, it is characterized in that, described drug-loading system is made up of the Cabazitaxel of massfraction to be the amphipathic nature block polymer of 85%-99.9% and massfraction be 0.1-15%, described amphipathic nature block polymer comprises hydrophilic segment and hydrophobic chain segment, hydrophilic segment is the polyoxyethylene glycol of molecular weight 100-6000 or the methoxy poly (ethylene glycol) of molecular weight 300-6000, hydrophobic chain segment is the polylactide of molecular weight 200-6000, hydrophobic chain segment carries out end-blocking with the group containing fluorenes first oxygen carboxyl and/or the amino acid containing benzene ring structure, the part by weight of polyoxyethylene glycol or methoxy poly (ethylene glycol) and polylactide is 0.1-30: 1.
2. drug-loading system according to claim 1, it is characterised in that, hydrophilic chain is the methoxy poly (ethylene glycol) of molecular weight 300-4000, and hydrophobic chain is the polylactide of molecular weight 300-5000; The part by weight of polyoxyethylene glycol or methoxy poly (ethylene glycol) and polylactide is 0.5-10: 1.
3. drug-loading system according to claim 1, it is characterised in that, hydrophilic chain is the methoxy poly (ethylene glycol) of molecular weight 600-3000, and hydrophobic chain is the polylactide of molecular weight 400-4000; The part by weight of polyoxyethylene glycol or methoxy poly (ethylene glycol) and polylactide is 0.5-10: 1.
4. according to the claim 1-3 described drug-loading system of arbitrary item, it is characterised in that, described drug-loading system is made up of the Cabazitaxel of massfraction to be the amphipathic nature block polymer of 90%-97% and massfraction be 3-10%; Described hydrophilic segment is molecular weight is the methoxy poly (ethylene glycol) of 1500-3000, hydrophobic chain segment is the polylactide of molecular weight 1000-2000, this hydrophobic chain segment carries out end-blocking with the group containing fluorenes first oxygen carboxyl and/or the amino acid containing benzene ring structure, and the part by weight of methoxy poly (ethylene glycol) and polylactide is 0.5-10: 1.
5. drug-loading system according to claim 1, it is characterized in that, the described group containing fluorenes first oxygen carboxyl is arbitrary or its combination in 6-fluorenylmethyloxycarbonyl amino, 6-fluorenylmethyloxycarbonyl amino-2-t-butoxycarbonyl amino caproic acid, 6-fluorenylmethyloxycarbonyl amino-2-benzyloxycarbonyl amino caproic acid.
6. a freeze-dried preparation containing drug-loading system described in claim 1, it is characterised in that, described freeze-dried preparation is made up of drug-loading system and freeze-drying auxiliary material, Cabazitaxel and freeze-drying auxiliary material weight ratio be 1: 0.1-50.
7. freeze-dried preparation according to claim 6, it is characterised in that, the weight ratio of described Cabazitaxel and freeze-drying auxiliary material is 1: 0.5-30.
8. freeze-dried preparation according to claim 6, it is characterised in that, the weight ratio of described Cabazitaxel and freeze-drying auxiliary material is 1: 1-10.
9. freeze-dried preparation according to claim 1, it is characterised in that, described freeze-drying auxiliary material is one or more combinations of lactose, N.F,USP MANNITOL, Xylitol, sorbyl alcohol, sucrose, trehalose, fructose, glucose, sodium alginate or gelatin.
10. the preparation method of drug-loading system described in a claim 1, it is characterised in that, comprise the following steps:
(1) polyoxyethylene glycol or methoxy poly (ethylene glycol) are joined in polymerization bottle, intensification is stirred and is vacuumized, rac-Lactide is added after cooling, stannous octoate, reactant high temperature polymerization after product methylene dichloride dissolves, and filters to obtain solid polyethylene glycol or methoxy poly (ethylene glycol) block polymer after precipitation;
(2) triethylamine is added after being dissolved by fluorenylmethyloxycarbonyl amino anhydrous tetrahydro furan, adding pivalyl chloride after cooling, have precipitation to generate, reactant continues reaction after heating up, after being filtered to remove impurity, evaporating solvent is to dry, obtains new penta acid anhydride that thick liquid is fluorenylmethyloxycarbonyl amino;
(3) polyoxyethylene glycol or methoxy poly (ethylene glycol) block polymer are dissolved in methylene dichloride, add triethylamine, 4-pyrollidinopyridine, after this solution is cooled, new penta acid anhydride of the fluorenylmethyloxycarbonyl amino of above-mentioned reaction gained is dissolved in methylene dichloride, after join in the dichloromethane solution of polyoxyethylene glycol or methoxy poly (ethylene glycol) block polymer, heat up after stirring, continue stirring reaction. Reactant precipitate after filter solid is the amino-terminated polyoxyethylene glycol of fluorenylmethyloxycarbonyl or methoxy poly (ethylene glycol)-polylactide block polymer;
(4) polymkeric substance and Cabazitaxel are made micellar solution.
11. preparation methods as claimed in claim 10, it is characterised in that, comprise the following steps:
(1) 10g methoxy poly (ethylene glycol) 2000 joins in polymerization bottle, intensification is stirred and is vacuumized, and adds rac-Lactide 11g, stannous octoate 5.5mg after cooling, reactant high temperature polymerization after product methylene dichloride dissolves, filter after ether sedimentation white solid both for mPEG-PLA block polymer;
(2) 11.7g6-fluorenylmethyloxycarbonyl amino-2-t-butoxycarbonyl amino caproic acid anhydrous tetrahydro furan adds 3-10mL triethylamine after dissolving, and adds pivalyl chloride 2-5mL after cooling, has precipitation to generate. Reactant continues reaction 1h after heating up, after being filtered to remove impurity, evaporating solvent is to dry, and obtaining thick liquid is 6-fluorenylmethyloxycarbonyl amino new penta acid anhydride of-2-t-butoxycarbonyl amino caproic acid;
(3) 20gmPEG-PLA is dissolved in methylene dichloride, add triethylamine 5mL, 4-pyrollidinopyridine 1.0g, after this solution is cooled, 6-fluorenylmethyloxycarbonyl amino new penta acid anhydride of-2-t-butoxycarbonyl amino caproic acid of above-mentioned reaction gained is dissolved in methylene dichloride, after join in the dichloromethane solution of mPEG-PLA, heat up after stirring reaction 1h, continue stirring reaction.Reactant filter after precipitating solid had both been the methoxy poly (ethylene glycol)-polylactide block polymer (mPEG-PLA-Lys (Fmoc)) of 6-fluorenylmethyloxycarbonyl amino-2-t-butoxycarbonyl amino caproic acid end-blocking;
(4) polymkeric substance and Cabazitaxel are made micellar solution.
12. preparation methods as claimed in claim 11, it is characterised in that, described drug-loading system is prepared into freeze-dried preparation, comprises the following steps:
By freeze-dried mixed to step (4) gained micellar solution and freeze-drying auxiliary material, obtained freeze-dried preparation.
13. preparation methods according to claim 10, it is characterised in that, there is the step of aseptically process before described step of freeze drying.
Drug-loading system described in 14. 1 kinds of claims 1 is in the application of the medicine of preparation treatment tumor disease.
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