CN103142482B - Preparation method for nanoparticle pharmaceutical composition, and nanoparticle pharmaceutical composition - Google Patents
Preparation method for nanoparticle pharmaceutical composition, and nanoparticle pharmaceutical composition Download PDFInfo
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Abstract
The invention provides a preparation method for a nanoparticle pharmaceutical composition. The method comprises the following steps of (1) mixing and emulsifying a first solution with an aqueous solution of hydrophilic small molecule drugs to obtain a first emulsion, wherein the first solution comprises an amphiphilic polymer and a first organic solvent; (2) mixing the first emulsion with an aqueous solution of a first surfactant, mixing the mixed materials with a second solution and an aqueous solution of a cationic polymer and emulsifying to obtain a second emulsion, wherein the second solution contains hydrophobic drugs and a second organic solvent; (3) mixing the second emulsion with an aqueous solution of a second surfactant, carrying out centrifugal separation to obtain a precipitate; and (4) dissolving the precipitate in water and mixing the dissolved precipitate with nucleic acid drugs, and separating to obtain the nanoparticle pharmaceutical composition. The invention also provides the nanoparticle pharmaceutical composition prepared by the above method.
Description
Technical field
The present invention relates to nanoparticle drug world, particularly, the nanoparticle pharmaceutical composition that relates to a kind of preparation method of nanoparticle pharmaceutical composition and prepared by the method.
Background technology
Nanometer medicine-carried system refers to that the particle diameter of medicine and nano-carrier formation, between the delivery system of 1-1000nm, comprises nanosphere, nanocapsule, nanoparticle and nanometer liposome etc.Nanometer medicine-carried system, compared with other medicines carrier, has significant advantage: (1) ultra micro small size, can pass through the blood capillary of human body minimum, and be difficult for being removed rapidly by phagocyte, extend the retention time in blood circulation; (2) arrive the reticuloendothelial system target sites such as concentrated liver, spleen, lung, bone marrow, lymph that distribute; (3) can penetrate tissue gap and by Cell uptake, be conducive to drug effect performance in Transdermal absorption and cell; (4) medicine can embedding or is bonded in inside nanoparticles, and also adsorbable or coupling is on its surface; (5) utilize the biodegradability of nano material itself, pH or temperature sensitivity etc., reach the effect that medicine control discharges; (6) improve the bioavailability of medicine and reduce toxic and side effects etc.
Nanoparticle is that one conventional in nanometer medicine-carried system is selected, and is typically used as the Polymer-supported medicine into pharmaceutical carrier, obtains nanoparticle pharmaceutical composition.But the nanoparticle pharmaceutical composition that the preparation method of current disclosed nanoparticle pharmaceutical composition obtains mostly is the nanoparticle pharmaceutical composition of a kind of medical compounds of load.As for hydrophilic small molecules medical compounds, mode by hydrophilic emulsifying loads on medical compounds on polymer, concrete grammar is: amphipathic nature polyalcohol is dissolved in organic solvent, and with the aqueous solution of hydrophilic small molecules medical compounds after under Ultrasonic Conditions, carry out emulsifying for the first time, material after emulsifying adds surfactant to carry out emulsifying for the second time for the first time, by the emulsion dispersion obtaining for the second time in aqueous surfactant solution and rotary evaporation remove organic solvent, after centrifugalize, obtain the nanoparticle compositions of load hydrophilic small molecules medical compounds.As for hydrophobic pharmaceutical compounds, mode by hydrophobicity emulsifying loads on medical compounds on amphipathic nature polyalcohol, concrete grammar is: the organic solution that is dissolved with medicine and amphipathic nature polyalcohol is slowly splashed into and contained in aqueous surfactant solution, after stirring certain hour, ultrasonic emulsification obtains emulsion, the emulsion rotary evaporation obtaining is removed to organic solvent, after centrifugalize, obtain the nanoparticle compositions of load hydrophobic pharmaceutical compounds.In the time of load multi-medicament compound, if respectively by the mode of hydrophilic emulsifying or the mode of hydrophobicity emulsifying by compound loaded multi-medicament on polymer, in the nanoparticle pharmaceutical composition obtaining, content ratio between each medical compounds is difficult to control, and has limited the raising of the result of use of nanoparticle pharmaceutical composition.
Therefore there is the unmanageable defect of content ratio between each medical compounds in existing nanoparticle pharmaceutical composition.
Summary of the invention
Have the unmanageable defect of content ratio between each medical compounds in order to overcome existing nanoparticle pharmaceutical composition, the invention provides a kind of preparation can be by the preparation method of the nanoparticle pharmaceutical composition of the content ratio between the each medical compounds of demand control and nanoparticle pharmaceutical composition prepared by the method.
The present inventor finds, different medical compoundss is due to hydrophilic and/or hydrophobicity difference, respectively by the mode of hydrophilic emulsifying or the mode of hydrophobicity emulsifying by compound loaded multi-medicament on amphipathic nature polyalcohol time, in the nanoparticle pharmaceutical composition obtaining, content ratio between each medical compounds can only be compared with close limit, and cannot adjust according to medication demand; But, if hydrophilic and/or the different medical compounds of hydrophobicity are loaded on described amphipathic nature polyalcohol by the multi-emulsion method with twice emulsifying step, just can adjust the content ratio between each medical compounds in the nanoparticle pharmaceutical composition obtaining according to medication demand easily, and due to mixing of cationic polymer, can realize the load of nucleic acid drug, obtain thus the present invention.
The invention provides a kind of preparation method of nanoparticle pharmaceutical composition, described nanoparticle pharmaceutical composition contains amphipathic nature polyalcohol, cationic polymer, hydrophobic drug, hydrophilic small molecules medicine and nucleic acid drug, and the method comprises the following steps:
(1) under Ultrasonic Radiation condition, make aqueous solution the emulsifying of the first solution and hydrophilic small molecules medicine, obtain the first emulsion;
Described the first solution contains described amphipathic nature polyalcohol and the first organic solvent; Described the first organic solvent is for can dissolve described amphipathic nature polyalcohol, but water insoluble, and under emulsification condition not with the organic solvent of described amphipathic nature polyalcohol, described hydrophobic drug, described hydrophilic small molecules medicine and described nucleic acid drug generation chemical reaction;
(2) by described the first emulsion and first surface activating agent aqueous solution, and by aqueous solution the emulsifying of mixed gained material and the second solution and cationic polymer, obtain the second emulsion;
Described the second solution contains described hydrophobic drug and the second organic solvent; Described the second organic solvent is for can dissolve described hydrophobic drug, but water insoluble, and under emulsification condition not with the organic solvent of described amphipathic nature polyalcohol, described hydrophobic drug, described hydrophilic small molecules medicine and described nucleic acid drug generation chemical reaction;
(3) by described the second emulsion and second surface activating agent aqueous solution, and remove described the first organic solvent and described the second organic solvent, then centrifugal and separation is precipitated thing.
(4) described precipitate is mixed with nucleic acid drug with aqueous solution dissolving is rear, separate and obtain nanoparticle pharmaceutical composition.
Method provided by the invention, can adjust and make in nanoparticle compositions provided by the invention according to medication demand easily, between the different medical compounds of hydrophilic and/or hydrophobicity and with the content ratio of nucleic acid drug.By above-mentioned adjustment, can utilize efficiently the peculiar advantage of nanometer medicine-carried system on administering mode, reach best compatibility of drugs result of use, especially can make being subject in the regional area of medicine target site, for example tumor occurs in site, and hydrophilic small molecules medicine, hydrophobic drug and nucleic acid drug compound reach best compatibility of drugs result of use.
The present invention also provides a kind of nanoparticle pharmaceutical composition, this nanoparticle pharmaceutical composition is prepared by said method, wherein, described hydrophobic drug is paclitaxel, described hydrophilic small molecules medicine is amycin, described nucleic acid drug is siRNA, described amphipathic nature polyalcohol is MPEG-PLA-ethanol copolymer, described cationic polymer is ε-poly-D-lysine, in described nanoparticle pharmaceutical composition, the weight ratio of described hydrophobic drug and described hydrophilic small molecules medicine is 1: 10000-10000: 1.
Experimental results show that, above-mentioned nanoparticle pharmaceutical composition provided by the invention, with be 1 by paclitaxel with the weight ratio of amycin by the nanoparticle pharmaceutical composition of only paclitaxel loaded nanoparticle pharmaceutical composition and a load amycin: after 0.9-1.1 mixes, compared with the pharmaceutical composition that obtains, there is better therapeutic effect for tumors such as melanoma, pulmonary carcinoma and hepatocarcinoma.
Other features and advantages of the present invention are described in detail the detailed description of the invention part subsequently.
Brief description of the drawings
Fig. 1 is the image of observing the nanoparticle pharmaceutical composition that embodiment 1 obtains under transmission electron microscope.
Fig. 2 is the grain-size graph that laser particle analyzer records the nanoparticle pharmaceutical composition that embodiment 1 obtains.
Detailed description of the invention
Below in conjunction with accompanying drawing, the specific embodiment of the present invention is elaborated.Should be understood that, detailed description of the invention described herein only, for description and interpretation the present invention, is not limited to the present invention.
The invention provides a kind of preparation method of nanoparticle pharmaceutical composition, described nanoparticle pharmaceutical composition contains amphipathic nature polyalcohol, cationic polymer, hydrophobic drug, hydrophilic small molecules medicine and nucleic acid drug, and the method comprises the following steps:
(1) under Ultrasonic Radiation condition, make aqueous solution the emulsifying of the first solution and hydrophilic small molecules medicine, obtain the first emulsion;
Described the first solution contains described amphipathic nature polyalcohol and the first organic solvent; Described the first organic solvent is for can dissolve described amphipathic nature polyalcohol, but water insoluble, and under emulsification condition not with the organic solvent of described amphipathic nature polyalcohol, described hydrophobic drug, described hydrophilic small molecules medicine and described nucleic acid drug generation chemical reaction;
(2) by described the first emulsion and first surface activating agent aqueous solution, and by aqueous solution the emulsifying of mixed gained material and the second solution and cationic polymer, obtain the second emulsion;
Described the second solution contains described hydrophobic drug and the second organic solvent; Described the second organic solvent is for can dissolve described hydrophobic drug, but water insoluble, and under emulsification condition not with the organic solvent of described amphipathic nature polyalcohol, described hydrophobic drug, described hydrophilic small molecules medicine and described nucleic acid drug generation chemical reaction;
(3) by described the second emulsion and second surface activating agent aqueous solution, and remove described the first organic solvent and described the second organic solvent, then centrifugal and separation is precipitated thing.
(4) described precipitate is mixed with nucleic acid drug with aqueous solution dissolving is rear, separate and obtain nanoparticle pharmaceutical composition.
Wherein, in step (1), the volume ratio of the aqueous solution of described the first solution and described hydrophilic small molecules medicine does not have special requirement, in order further to improve the result of use of described nanoparticle pharmaceutical composition, under preferable case, the volume ratio of the aqueous solution of described the first solution and described hydrophilic small molecules medicine is 1: 1-1: 10; More preferably 1: 2-1: 6, most preferably be 1: 4.
According to the present invention, in described the first solution, with respect to first organic solvent of every milliliter, the content of described amphipathic nature polyalcohol does not have special requirement, in order further to improve the result of use of described nanoparticle pharmaceutical composition, under preferable case, in described the first solution, with respect to first organic solvent of every milliliter, the content of described amphipathic nature polyalcohol is 5-100 milligram; More preferably 20-40 milligram.
According to the present invention, in the aqueous solution of described hydrophilic small molecules medicine, with respect to the water of every milliliter, the content of described hydrophilic small molecules medicine does not have special requirement, in order further to improve the result of use of described nanoparticle pharmaceutical composition, under preferable case, in the aqueous solution of described hydrophilic small molecules medicine, with respect to the water of every milliliter, the content of described hydrophilic small molecules medicine is 1-100 milligram; More preferably 10-50 milligram.
According to the present invention, in step (1), the selection of described amphipathic nature polyalcohol does not have special requirement, can be the selection of nanoparticle pharmaceutical composition preparation field routine, for example, described amphipathic nature polyalcohol is the polymer that simultaneously contains hydrophilic radical and hydrophobic group; Described hydrophilic radical can be one or more in polyethylene group, polyoxyethylene group and polyisobutylene acid groups etc., and described hydrophobic group can be medium one or more of polyoxyethylene group, polylactic acid-glycollic acid group, polystyrene group and polycaprolactone group.It should be noted that, the arrangement mode of the repetitive of described amphipathic nature polyalcohol is requirement especially not, as long as being used to form nanoparticle pharmaceutical composition, can be for example one or more alternately, in block, random or grafting, also can be obtained by the monomer homopolymerization that contains hydrophilic radical and hydrophobic group; The molecular weight of described amphipathic nature polyalcohol does not have special requirement, and as long as being used to form nanoparticle pharmaceutical composition, the weight average molecular weight that the method that for example described amphipathic nature polyalcohol specifies according to SHT 1759-2007 records can be 10
4-10
5.
According to the present invention, in order to make described nanoparticle pharmaceutical composition can also have more special Targeting Performance and/or imaging performance, under preferable case, in step (1), described amphipathic nature polyalcohol also contains targeting group and/or imaging group.
Wherein, the selection of described targeting group and/or imaging group does not have special requirement, it can be the selection of nanoparticle pharmaceutical composition preparation field routine, in order further to improve the result of use of described nanoparticle pharmaceutical composition, further, under preferable case, described targeting group is one or more in folic acid group, essence-Gan-aspartic acid motif cyclic peptide (RGD) group and transferrins group etc.; Described imaging group is quantum dot group and/or fluorescent dye group.For example, described quantum dot group can be one or more in the nano materials such as golden quantum dot group, cadmium quantum dot group and palladium quantum dot group; Described fluorescent dye group can be one or more in Fluorescein isothiocyanate group, eosin disodium group and acid red 87 group.
Wherein, the not requirement especially of the content of described targeting group and/or imaging group can be the selection of nanoparticle pharmaceutical composition preparation field routine, for example, with respect to the hydrophilic radical of every mole, the content of described targeting group and/or imaging group can be 0.2-1 mole.
A preferred embodiment of the invention, in step (1), described amphipathic nature polyalcohol is one or more in MPEG-PLA-glycolic, poly(ethylene oxide)-poly(propylene oxide), Polyethylene Glycol and poly (l-lactic acid).
Wherein, the hydrophilic group of described MPEG-PLA-glycolic is poly glycol monomethyl ether group, hydrophobic group is polylactic acid-glycollic acid group, the arrangement mode of the repetitive of described MPEG-PLA-ethanol copolymer is block, wherein, mol ratio between ethylene glycol unit and lactic acid-ethanol unit can be 1: 1-1: 8, and the weight average molecular weight that the method that described MPEG-PLA-ethanol copolymer specifies according to SHT 1759-2007 records can be 10
4-10
5.MPEG-PLA-the ethanol copolymer that meets above-mentioned requirements can be by commercially available.
According to the present invention, the selection of described hydrophilic small molecules medicine does not have special requirement, can select by the dissolubility in water according to desired drug effect and medicine, the molecular weight of described hydrophilic small molecules medicine can be below 1000Da, for example, can be chosen in medicine that the dissolubility in water is greater than 1g/100g as hydrophilic small molecules medicine, under preferable case, described hydrophilic small molecules medicine is one or more in amycin, mitoxantrone, daunorubicin and epirubicin.
According to the present invention, the selection of described the first organic solvent does not have special requirement, it can be the selection of nanoparticle pharmaceutical composition preparation field routine, in order further to improve the result of use of described nanoparticle pharmaceutical composition and to reduce costs, under preferable case, described the first organic solvent is one or more in dichloromethane, dimethyl sulfoxide, oxolane, acetone, espeleton, dichloro-benzenes and methyl isopropyl ketone.
According to the present invention, in step (1), the condition of described emulsifying does not have special requirement, it can be the selection of nanoparticle pharmaceutical composition preparation field routine, in order further to improve the result of use of described nanoparticle pharmaceutical composition, under preferable case, in step (1), the condition of described emulsifying comprises: the frequency of Ultrasonic Radiation is 20-25kHz, more preferably 22-23kHz; With respect to every milliliter of material of accepting Ultrasonic Radiation, power is 19-190W, more preferably 28.5-57W; The temperature of emulsifying is 1-99 DEG C, more preferably 20-30 DEG C; The time of emulsifying is 1-30 minute, more preferably 3-10 minute.
Wherein, in step (1), the mode of described mixing does not have special requirement, it can be the selection of nanoparticle pharmaceutical composition preparation field routine, in order further to improve the result of use of described nanoparticle pharmaceutical composition, under preferable case, in step (1), the mode of described mixing comprises: the aqueous solution of described hydrophilic small molecules medicine is joined in the first solution and mixed.
According to the present invention, in step (2), the consumption of the aqueous solution of the consumption of described first surface activating agent aqueous solution, described the second solution and cationic polymer does not have special requirement, in order further to improve the result of use of described nanoparticle pharmaceutical composition, under preferable case, in step (2), with respect to the first emulsion of 1 volume, the consumption of described first surface activating agent aqueous solution is 0.5-2 volume, more preferably 0.8-1 volume; The consumption of described the second solution is 0.1-1 volume, more preferably 0.4-0.6 volume; The consumption of the aqueous solution of described cationic polymer is 0.1-1 volume, more preferably 0.4-0.6 volume.
Wherein, in step (2), in described first surface activating agent aqueous solution, with respect to the water of every milliliter, the content of described first surface activating agent does not have special requirement, it can be the selection of nanoparticle pharmaceutical composition preparation field routine, in order further to improve the result of use of described nanoparticle pharmaceutical composition, under preferable case, in step (2), in described first surface activating agent aqueous solution, the content of described first surface activating agent is 0.6-6 % by weight, more preferably 2-4 % by weight.
Wherein, in step (2), in described the second solution, with respect to second organic solvent of every milliliter, the content of described hydrophobic drug does not have special requirement, in order further to improve the result of use of described nanoparticle pharmaceutical composition, under preferable case, in step (2), in described the second solution, with respect to second organic solvent of every milliliter, the content of described hydrophobic drug can be 0.1 milligram to reach capacity in described the second organic solvent content of concentration of described hydrophobic drug, more preferably 1-10 milligram.
Wherein, in step (2), in the aqueous solution of described cationic polymer, the content of described cationic polymer does not have special requirement, in order further to improve the result of use of described nanoparticle pharmaceutical composition, under preferable case, in step (2), in described the 4th solution, the content of described cationic polymer can be 1 milligram to reach capacity in the institute's aqueous solution content of concentration of described cationic polymer, more preferably 20-40 milligram.
According to the present invention, in step (2), the effect of described first surface activating agent is the stability that improves the second emulsion, thereby can improve the stability of nanoparticle pharmaceutical composition, its selection does not have special requirement, it can be the selection of nanoparticle pharmaceutical composition preparation field routine, in order further to improve the result of use of described nanoparticle pharmaceutical composition, under preferable case, in step (2), described first surface activating agent is one or more in polyvinyl alcohol, POLYSORBATE 80, span and dodecyl sodium sulfate.
Wherein, the selection of described polyvinyl alcohol does not have special requirement, can be the selection of nanoparticle pharmaceutical composition preparation field routine, and for example, described polyvinyl alcohol can be pharmaceutical grade polyvinyl alcohol; The saponification degree of described polyvinyl alcohol can be 70-95mol%; The viscosity of described polyvinyl alcohol at 25 DEG C can be 500-900mPas.The polyvinyl alcohol that meets above-mentioned requirements can be by commercially available, the product that for example can to buy from the trade mark of Chemical Reagent Co., Ltd., Sinopharm Group be PVA-124.
Wherein, the selection of described hydrophobic drug does not have special requirement, can select by the dissolubility in water according to desired drug effect and medicine, for example, can be chosen in medicine that the dissolubility in water is less than 1g/100g as hydrophobic drug, under preferable case, described hydrophobic drug is one or more in paclitaxel, camptothecine, harringtonine and vinorelbine.
Wherein, the selection of described cationic polymer does not have special requirement, can select according to desired nanoparticle surface electromotive force, for example, can be chosen in the cationic polymer for positive potential in water, under preferable case, described cationic polymer is one or more in poly-D-lysine, chitosan and Polyetherimide-amine type dendrimer.Described poly-D-lysine is preferably ε-poly-D-lysine.
Wherein, in step (2), the condition of described emulsifying does not have special requirement, can select by the dissolubility in water according to desired drug effect and medicine, in order further to improve the result of use of described nanoparticle pharmaceutical composition, under preferable case, in step (2), the condition of described emulsifying comprises: the frequency of Ultrasonic Radiation is 20-25kHz, more preferably 22-23kHz; With respect to every milliliter of material of accepting Ultrasonic Radiation, power is 19-190W, more preferably 28.5-57W; The temperature of emulsifying is 1-99 DEG C, more preferably 20-30 DEG C; The time of emulsifying is 1-30 minute, more preferably 3-10 minute.
Wherein, in step (2), by in the process of described the first emulsion and described first surface activating agent aqueous solution, the mode of described mixing does not have special requirement, it can be the selection of nanoparticle pharmaceutical composition preparation field routine, in order further to improve the result of use of described nanoparticle pharmaceutical composition, under preferable case, in step (2), by in the process of described the first emulsion and described first surface activating agent aqueous solution, the mode of described mixing comprises: under the stirring condition of turn at 100-1200/min, described the first emulsion is joined in described first surface activating agent aqueous solution and mixed, and the speed that described the first emulsion adds is 0.1-3ml/min.
Wherein, in step (2), to as above mix in the aqueous solution emulsification of the mixed material of gained and described the second solution and described cationic polymer, the mode of described mixing does not have special requirement, it can be the selection of nanoparticle pharmaceutical composition preparation field routine, in order further to improve the result of use of described nanoparticle pharmaceutical composition, under preferable case, in step (2), by in the aqueous solution emulsification of mixed gained material and described the second solution and described cationic polymer, the mode of described mixing comprises: under the stirring condition of turn at 1000-1200/min, the aqueous solution of described the second solution and described cationic polymer is joined and as above mixed in the mixed material of gained, and the speed that described the second solution adds is 0.1-1ml/min.
According to the present invention, in step (3), with respect to described second emulsion of 1 volume, the consumption of described second surface activating agent aqueous solution does not have special requirement, it can be the selection of nanoparticle pharmaceutical composition preparation field routine, in order further to improve the result of use of described nanoparticle pharmaceutical composition, under preferable case, in step (3), with respect to described second emulsion of 1 volume, the consumption of described second surface activating agent aqueous solution is 2-20 volume, more preferably 10-15 volume.
Wherein, in step (3), in described second surface activating agent aqueous solution, with respect to the water of every milliliter, the content of described second surface activating agent does not have special requirement, it can be the selection of nanoparticle pharmaceutical composition preparation field routine, in order further to improve the result of use of described nanoparticle pharmaceutical composition, under preferable case, in step (3), in described second surface activating agent aqueous solution, the content of described second surface activating agent is 0.3-3 % by weight, more preferably 0.6-1 % by weight.
Wherein, in step (3), the effect of described second surface activating agent is to improve described sedimentary stability, thereby can improve the stability of nanoparticle pharmaceutical composition, its selection does not have special requirement, it can be the selection of nanoparticle pharmaceutical composition preparation field routine, in order further to improve the result of use of described nanoparticle pharmaceutical composition, under preferable case, in step (3), described second surface activating agent is one or more in polyvinyl alcohol, POLYSORBATE 80, span and dodecyl sodium sulfate.
Wherein, the selection of described polyvinyl alcohol does not have special requirement, can be the selection of nanoparticle pharmaceutical composition preparation field routine, and for example, described polyvinyl alcohol can be pharmaceutical grade polyvinyl alcohol; The saponification degree of described polyvinyl alcohol can be 70-95mol%; The viscosity of described polyvinyl alcohol at 25 DEG C can be 500-900mPas.The polyvinyl alcohol that meets above-mentioned requirements can be by commercially available, the product that for example can to buy from the trade mark of Chemical Reagent Co., Ltd., Sinopharm Group be PVA-124.
Wherein, in step (3), by in the process of described the second emulsion and second surface activating agent aqueous solution, the mode of described mixing does not have special requirement, it can be the selection of nanoparticle pharmaceutical composition preparation field routine, in order further to improve the result of use of described nanoparticle pharmaceutical composition, under preferable case, in step (3), by in the process of described the second emulsion and second surface activating agent aqueous solution, the mode of described mixing comprises: under the stirring condition of turn at 100-1200/min, described the second emulsion is joined in described second surface activating agent aqueous solution, and the speed that described the second emulsion adds is 0.1-3ml/min, add after described the second emulsion, can under the stirring condition of turn at 100-1200/min, maintain 5-100min.
Wherein, in step (3), remove in the process of described the first organic solvent and the second organic solvent, described method of removing described the first organic solvent and the second organic solvent does not have special requirement, for example, can remove described the first organic solvent and the second organic solvent by the mode of rotary evaporation.
Wherein, in step (3), it can be 5000-15000g that centrifugal and separation is precipitated speed centrifugal in the process of thing.
According to the present invention, wherein, in step (4), with respect to the described precipitate of 1 weight portion, the consumption of water can be 1-1000 weight portion, is preferably 10-100 weight portion.
Wherein, in step (4), with respect to the described precipitate of 1 weight portion, the consumption of described nucleic acid drug can be 0.001-1 weight portion, is preferably 0.1-0.2 weight portion.
Wherein, described nucleic acid drug comprises the various medicines taking nucleic acid as effective ingredient, and for example DNA medicine and RNA medicine, be preferably siRNA medicine.
Wherein, in step (4), in the process that described precipitate is dissolved with aqueous solution and the process of mixing with nucleic acid drug subsequently, the mode of dissolving and the mode of mixing do not have special requirement, in order further to improve the result of use of described nanoparticle pharmaceutical composition, under preferable case, in step (4), the mode that just described precipitate mixes with nucleic acid drug after dissolving with aqueous solution comprises: under the stirring condition of turn at 100-1200/min, described precipitate and described nucleic acid drug are added to the water, then, can under the stirring condition of turn at 100-1200/min, maintain 5-100min.
Wherein, in step (4), separation obtains in the process of described nanoparticle pharmaceutical composition, the method of described separation does not have special requirement, for example, can separate by the method for centrifugal and collecting precipitation (obtain be precipitated as described nanoparticle pharmaceutical composition), centrifugal speed can be 5000-15000g.
The present invention also provides a kind of nanoparticle pharmaceutical composition, this nanoparticle pharmaceutical composition is prepared by said method, wherein, described hydrophobic drug is paclitaxel, described hydrophilic small molecules medicine is amycin, described nucleic acid drug is siRNA, described amphipathic nature polyalcohol is MPEG-PLA-ethanol copolymer, described cationic polymer is ε-poly-D-lysine, in described nanoparticle pharmaceutical composition, the weight ratio of described hydrophobic drug and described hydrophilic small molecules medicine is 1: 10000-10000: 1.
Wherein, under preferable case, the weight ratio of described hydrophobic drug and described hydrophilic small molecules medicine is 1: 1000-1000: 1.
Wherein, described nucleic acid drug can be in required ratio and nanoparticle mixed phase the described precipitate for 1 weight portion, the consumption of described nucleic acid drug can be 0.001-1 weight portion, is preferably 0.01-0.2 weight portion.
While experiment showed, described nanoparticle pharmaceutical composition for mice, calculating with the amount of amycin, is 0.1-10mg/kg body weight through the effective dose of intravenously administrable.
In the present invention, the volume of gas and liquid is at 20 DEG C, the numerical value that standard atmosphere is depressed.
Below, further describe the present invention by embodiment, but scope of the present invention is not limited in following examples.
Embodiment 1
The present embodiment is prepared nanoparticle pharmaceutical composition according to following steps.
(1) by amphipathic nature polyalcohol (MPEG-PLA-glycolic of 20mg, PLGA-PEG, purchased from Dai Gang bio tech ltd, Jinan, the hydrophilic group of this MPEG-PLA-ethanol copolymer is poly glycol monomethyl ether group, hydrophobic group is polylactic acid-glycollic acid group, the arrangement mode of the repetitive of described MPEG-PLA-ethanol copolymer is block, wherein, mol ratio between ethylene glycol unit and lactic acid-ethanol unit is 1: 1-1: 8, the weight average molecular weight that the method specifying according to SHT 1759-2007 records is 10
4-1.5 × 10
5) be dissolved in 1ml the first organic solvent (dichloromethane), obtain the first solution.The hydrophilic small molecules medicine of 0.6mg (amycin, purchased from Beijing Hua Feng Science and Technology Ltd., the trade mark is HF090516) is dissolved in 1ml water, obtains the aqueous solution of hydrophilic small molecules medicine.The aqueous solution of the hydrophilic small molecules medicine of 250 μ l is added in the first solution of 1ml, obtain mixed material.At 25 DEG C, be 28.5W and the frequency Ultrasonic Radiation that is 23kHz after 3 minutes by the obtained above mixed material of 1ml with power, obtain the first emulsion.
(2) by first surface activating agent (polyvinyl alcohol, purchased from Chemical Reagent Co., Ltd., Sinopharm Group, the trade mark is PVA-124, saponification degree is 85mol%; Viscosity at 25 DEG C is 700mPas) be formulated as the aqueous solution of concentration 2 % by weight, obtain first surface activating agent aqueous solution.Under the stirring condition of turn 1000/min, the first emulsion that 1ml step (1) is obtained joins in 1ml first surface activating agent aqueous solution with the speed of 1ml/min, obtains mixed material.0.125mg hydrophobic drug (paclitaxel, purchased from Beijing Nuorui Medical Tech. Co., Ltd., the trade mark is 090328) is dissolved in 1ml the second organic solvent (dichloromethane), obtains the second solution.The cationic polymer of 20mg (ε-poly-D-lysine, purchased from Yinxiang Biological Engineering Co., Ltd., Zhejiang Prov) is dissolved in 0.1ml aqueous solution, obtains the aqueous solution of cationic polymer.Under the stirring condition of turn 1000/min, the aqueous solution of the cationic polymer obtained above of the second solution obtained above of 0.25ml and 0.1ml is added in mixed material obtained above with the speed of 1ml/min respectively, obtain for ultrasonic material.At 25 DEG C, by obtained above 1ml be 57W and the frequency Ultrasonic Radiation that is 23kHz after 5 minutes for ultrasonic material with power, obtain the second emulsion.
(3) by second surface activating agent (polyvinyl alcohol, purchased from Chemical Reagent Co., Ltd., Sinopharm Group, the trade mark is PVA-124, saponification degree is 85mol%; Viscosity at 25 DEG C is 700mPas) be formulated as the aqueous solution of concentration 0.6%, obtain second surface activating agent aqueous solution.Under the stirring condition of turn 800/min, the second emulsion that 1ml step (2) is obtained joins in the above-mentioned second surface activating agent aqueous solution of 10ml with the speed of 2ml/min, under the stirring condition of then turn at 600-800/min, maintains 10min.At room temperature DEG C, rotary evaporation is removed the first organic solvent (dichloromethane) and the second organic solvent (dichloromethane), obtains revolving the product after steaming.The product revolving after steaming after centrifugal 10 minutes, is collected and is precipitated thing under the centrifugal speed of 13000g.
(4) precipitate obtaining (10mg left and right) is used after the water dissolution of 2ml, add the nucleic acid drug (siRNA of 0.05mg, positive-sense strand sequence is 5 '-GAAUUAACCCUUGGUGAAUTT-3 ' (SEQ ID NO:1), antisense strand sequence is 5 '-AUUCACCAAGGGUUAAUUCTT-3 ' (SEQ ID NO:2), purchased from the English Weihe River prompt base (Shanghai) trade Co., Ltd, this siRNA can reticent anti-apoptotic proteins survivin gene expression, thereby the growth of inhibition tumor cell), vibration mixes rear room temperature and places 30 minutes, under the centrifugal speed of 13000g after centrifugal 10 minutes, collecting precipitation, obtain nanoparticle pharmaceutical composition.
According to document (Ashlynn L.Z.Lee, Deng. biomaterial (Biomaterials) 2009,30, method 919-927), under transmission electron microscope, observe the nanoparticle pharmaceutical composition that the present embodiment obtains, result as shown in Figure 1, illustrates that the size of the nanoparticle pharmaceutical composition obtaining is about 200nm.
According to document (Ashlynn L.Z.Lee, Deng. biomaterial (Biomaterials) 2009,30, method 919-927), the mean diameter of utilizing laser particle analyzer to record the nanoparticle pharmaceutical composition that the present embodiment obtains is 200.63 ± 12.36nm (Fig. 2), dispersion is that 0.131, zeta current potential is 29.6 ± 0.14, shows that the stability of this nanoparticle pharmaceutical composition is better.
According to document (Ashlynn L.Z.Lee, Deng. biomaterial (Biomaterials) 2009,30, method 919-927), after the nanoparticle pharmaceutical composition lyophilization that the present embodiment is obtained, dissolve with acetonitrile, (standard substance are purchased from Agilent Technologies to utilize high performance liquid chromatography detection, trade mark Agilent 1200 series), can find out that nanoparticle pharmaceutical composition that the present embodiment obtains has the characteristic peak of amycin and paclitaxel simultaneously.And record in the nanoparticle pharmaceutical composition that the present embodiment obtains and contain siRNA by ultraviolet spectrophotometer method, illustrate in the nanoparticle pharmaceutical composition that amycin and paclitaxel obtain at the present embodiment and exist simultaneously, and in the nanoparticle pharmaceutical composition that the present embodiment of every gram obtains, the content of amycin is 0.15mg, the content of paclitaxel is 0.075mg, and the content of siRNA is 0.045mg.
Comparative example 1
This comparative example is prepared nanoparticle pharmaceutical composition according to the method for embodiment 1, difference is: by amphipathic nature polyalcohol (MPEG-PLA-ethanol copolymer of 20mg, PLGA-PEG, purchased from Dai Gang bio tech ltd, Jinan) be dissolved in 1ml water, the solution obtaining is as the first solution, and water is replaced the first organic solvent (dichloromethane).
According to the measuring method identical with embodiment 1, the particle diameter that utilizes laser particle analyzer to record the nanoparticle pharmaceutical composition that this comparative example obtains is 300nm; The content that records amycin in the nanoparticle pharmaceutical composition that this comparative example of every gram obtains is 0.05g, and the content of paclitaxel is 0g, and the content of siRNA is 0g.
Comparative example 2
Prepare nanoparticle pharmaceutical composition according to following steps.
(1) by amphipathic nature polyalcohol MPEG-PLA-ethanol copolymer of 20mg, PLGA-PEG, purchased from Dai Gang bio tech ltd, Jinan) be dissolved in 1ml the first organic solvent (dichloromethane), obtain the first solution.The hydrophilic small molecules medicine of 0.6mg (amycin, purchased from Beijing Hua Feng Science and Technology Ltd., the trade mark is HF090516) is dissolved in 1ml water, and the solution obtaining is as the aqueous solution of hydrophilic small molecules medicine.The aqueous solution that first surface activating agent (polyvinyl alcohol, purchased from Chemical Reagent Co., Ltd., Sinopharm Group, the trade mark is PVA-124) is formulated as to concentration 2 % by weight, obtains first surface activating agent aqueous solution.0.125mg hydrophobic drug (paclitaxel, purchased from Beijing Nuorui Medical Tech. Co., Ltd., the trade mark is 090328) is dissolved in 1ml the second organic solvent (dichloromethane), obtains the second solution.
Under the stirring condition of turn 1000/min, by the aqueous solution of the first solution obtained above, the second solution obtained above, hydrophilic small molecules medicine obtained above, first surface activating agent aqueous solution obtained above, obtain for ultrasonic material.At 25 DEG C, by the Ultrasonic Radiation that is first 23kHz by the frequency of 28.5W for ultrasonic material obtained above 1ml after 3 minutes, then the Ultrasonic Radiation that is 23kHz by the frequency of 57W 3 minutes, obtain the second emulsion.
(2) second surface activating agent (polyvinyl alcohol, purchased from Chemical Reagent Co., Ltd., Sinopharm Group, the trade mark is PVA-124) is formulated as to the aqueous solution of concentration 0.6 % by weight, obtains second surface activating agent aqueous solution.Under the stirring condition of turn 1000/min, the second emulsion that 1ml step (1) is obtained joins in the above-mentioned second surface activating agent aqueous solution of 10ml with the speed of 1ml/min, under the stirring condition of then turn at 600-800/min, maintains 10min.At room temperature, rotary evaporation 10min, removes the first organic solvent (dichloromethane) and the second organic solvent (dichloromethane), obtains revolving the product after steaming.The product revolving after steaming after centrifugal 10 minutes, is collected and is precipitated thing under the centrifugal speed of 13000g.
(4) precipitate obtaining (10mg left and right) is used after the water dissolution of 2ml, add the nucleic acid drug (siRNA of 0.05mg, in the same manner as in Example 1), vibration mixes rear room temperature and places 30 minutes, under the centrifugal speed of 13000g after centrifugal 10 minutes, collecting precipitation, obtains nanoparticle pharmaceutical composition.
According to the measuring method identical with embodiment 1, the particle diameter that utilizes laser particle analyzer to record the nanoparticle pharmaceutical composition that this comparative example obtains is 300nm; The content that records amycin in the nanoparticle pharmaceutical composition that this comparative example of every gram obtains is 0g, and the content of paclitaxel is 0.02g, and the content of siRNA is 0g.
Embodiment 2
The present embodiment is prepared nanoparticle pharmaceutical composition according to the method for embodiment 1, difference is, in the aqueous solution of described hydrophilic small molecules medicine, the concentration of amycin is 0.4mg/ml, and in described the second solution, the concentration of paclitaxel is 10mg/ml, and the consumption of described nucleic acid drug is 0.05mg.
According to the measuring method identical with embodiment 1, the particle diameter that utilizes laser particle analyzer to record the nanoparticle pharmaceutical composition that the present embodiment obtains is about 200nm; The content that records amycin in the nanoparticle pharmaceutical composition that the present embodiment of every gram obtains is 0.0005g, and the content of paclitaxel is 0.5g, and the content of siRNA is 0.000045g.
Comparative example 3
This comparative example is prepared nanoparticle pharmaceutical composition according to the method for comparative example 2, difference is, in the aqueous solution of described hydrophilic small molecules medicine, the concentration of amycin is 0.4mg/ml, and in described the second solution, the concentration of paclitaxel is 10mg/ml, and the consumption of described nucleic acid drug is 0.05mg.
According to the measuring method identical with embodiment 1, the particle diameter that utilizes laser particle analyzer to record the nanoparticle pharmaceutical composition that this comparative example obtains is 300nm; The content that records amycin in the nanoparticle pharmaceutical composition that this comparative example of every gram obtains is 0g, and the content of paclitaxel is 0.1g, and the content of siRNA is 0g.
Embodiment 3
The present embodiment is prepared nanoparticle pharmaceutical composition according to the method for embodiment 1, difference is, in the aqueous solution of described hydrophilic small molecules medicine, the concentration of amycin is 10mg/ml, and in described the second solution, the concentration of paclitaxel is 0.4mg/ml, and the consumption of described nucleic acid drug is 0.05mg.
According to the measuring method identical with embodiment 1, the particle diameter that utilizes laser particle analyzer to record the nanoparticle pharmaceutical composition that the present embodiment obtains is 260nm; The content that records amycin in the nanoparticle pharmaceutical composition that the present embodiment of every gram obtains is 0.47g, and the content of paclitaxel is 0.00047g, and the content of siRNA is 0.000045g.
Comparative example 4
This comparative example is prepared nanoparticle pharmaceutical composition according to the method for comparative example 2, difference is, in the aqueous solution of described hydrophilic small molecules medicine, the concentration of amycin is 10mg/ml, and in described the second solution, the concentration of paclitaxel is 0.4mg/ml, and the consumption of described nucleic acid drug is 0.05mg.
According to the measuring method identical with embodiment 1, the particle diameter that utilizes laser particle analyzer to record the nanoparticle pharmaceutical composition that this comparative example obtains is 300nm; The content that records amycin in the nanoparticle pharmaceutical composition that this comparative example of every gram obtains is 0g, and the content of paclitaxel is 0.1g, and the content of siRNA is 0g.
Comparative example 5
This comparative example is prepared nanoparticle pharmaceutical composition according to the method identical with comparative example 2, difference is, do not use the second solution, and in the aqueous solution of hydrophilic small molecules medicine, the use amount of amycin is 3.5mg, the use amount of nucleic acid drug (siRNA, in the same manner as in Example 1) is 0.1mg.
According to the measuring method identical with embodiment 1, the particle diameter that utilizes laser particle analyzer to record the nanoparticle pharmaceutical composition that this comparative example obtains is 300nm; The content that records amycin in the nanoparticle pharmaceutical composition that this comparative example of every gram obtains is 0.3g, and the content of paclitaxel is 0g, and the content of siRNA is 0.09mg.
Comparative example 6
This comparative example is prepared nanoparticle pharmaceutical composition according to the method identical with comparative example 2, and difference is do not use the aqueous solution of hydrophilic small molecules medicine, and in the second solution, the use amount of paclitaxel to be 2mg.
According to the measuring method identical with embodiment 1, the particle diameter that utilizes laser particle analyzer to record the nanoparticle pharmaceutical composition that this comparative example obtains is 300nm; The content that records amycin in the nanoparticle pharmaceutical composition that this comparative example of every gram obtains is 0g, and the content of paclitaxel is 0.15g.
Test implementation example 1
In this test implementation example, embodiment 1 is made to the nanoparticle compositions that is loaded with multi-medicament as Experimental agents; The nanoparticle medicine mixed in equal amounts that the nanoparticle pharmaceutical composition that comparative example 5 is obtained and comparative example 6 obtain, the content that obtains amycin is 15 % by weight, the content of paclitaxel is the nanoparticle pharmaceutical composition mixture of 7.5 % by weight.By this nanoparticle pharmaceutical composition mixture medicine in contrast.
Culture dish 10 by melanoma cell (B16 cell line, purchased from ATCC, is numbered B16-F10) with every 35mm internal diameter
3individual density is inoculated in respectively in two culture dishs of the DMEM culture medium that contains 2ml (purchased from Gibco, trade mark sh30022.01B, and containing the hyclone of 10 volume %), at the CO of 37 DEG C and 5 volume %
2under concentration, cultivate after 24 hours, sucking-off culture medium, the culture medium that 2ml is dissolved with to Experimental agents is (in amycin, drug level is 0.17 μ mol/ml) add wherein in a culture dish, the culture medium that 2ml is dissolved with to control drug is (in amycin, drug level is 0.17 μ mol/ml) add in another culture dish, by above-mentioned 2 culture dishs at 37 DEG C and 5% CO
2under concentration, hatch after 24 hours, detect different time sections Level of Apoptosis according to the CCK-8 method described in document (Qinghua Miao, etc., biomaterial (Biomaterials), 2010,31 (28): 7364-75).Can find, use in the culture dish of Experimental agents, cytoactive is 90.2 ± 1.2 (4h), 31.88 ± 2% (8h), 16.3 ± 3.2% (12h), 6.1 ± 0.05% (24h); Use in the culture dish of control drug, cytoactive is 96.2 ± 2.1% (4h), 53.63 ± 1.3% (8h), 42.5 ± 3.4% (12h), 34.43 ± 1.5 (24h).The above results illustrative experiment medicine has stronger short apoptosis effect to melanoma cell.
Use melanoma cell (B16 cell line, purchased from ATCC, numbering B16-F10) and C57 mice (C57 system, purchased from Beijing company of dimension tonneau China), according to document, (Fariyal Ahmed, learns (Molecular Pharmaceutics) etc. molecular medicine, 3, method 340-350) is prepared tumor bearing nude mice, treats that tumor grows to 0.5cm
2when size, tumor bearing nude mice is divided into experimental group tumor bearing nude mice and matched group tumor bearing nude mice at random, according to the method in document (the same), through tail vein, the phosphate buffer (in amycin, dosage is 3mg/kg body weight) that is dissolved with Experimental agents is administered to experimental group tumor bearing nude mice; The phosphate buffer (in amycin, dosage is 3mg/kg body weight) that is dissolved with control drug is administered to matched group tumor bearing nude mice.After one week, according to document, (Fariyal Ahmed, learns (Molecular Pharmaceutics) etc. molecular medicine, 3, method 340-350) is measured the size of tumor, wherein, and the tumor size average out to 452mm of experimental group tumor bearing nude mice
3, the tumor size average out to 8462mm of matched group tumor bearing nude mice
3.The above results explanation illustrative experiment medicine has stronger inhibition to melanoma.
Below describe by reference to the accompanying drawings the preferred embodiment of the present invention in detail; but; the present invention is not limited to the detail in above-mentioned embodiment; within the scope of technical conceive of the present invention; can carry out multiple simple variant to technical scheme of the present invention, these simple variant all belong to protection scope of the present invention.
Claims (9)
1. a preparation method for nanoparticle pharmaceutical composition, described nanoparticle pharmaceutical composition contains amphipathic nature polyalcohol, cationic polymer, hydrophobic drug, hydrophilic small molecules medicine and nucleic acid drug, and the method comprises the following steps:
(1) under Ultrasonic Radiation condition, make aqueous solution the emulsifying of the first solution and hydrophilic small molecules medicine, obtain the first emulsion;
Described the first solution contains described amphipathic nature polyalcohol and the first organic solvent; Described the first organic solvent is for can dissolve described amphipathic nature polyalcohol, but water-fast organic solvent;
(2) by described the first emulsion and first surface activating agent aqueous solution, and by aqueous solution the emulsifying of mixed gained material and the second solution and cationic polymer, obtain the second emulsion;
Described the second solution contains described hydrophobic drug and the second organic solvent; Described the second organic solvent is for can dissolve described hydrophobic drug, but water-fast organic solvent;
(3) by described the second emulsion and second surface activating agent aqueous solution, and remove described the first organic solvent and described the second organic solvent, then centrifugal and separation is precipitated thing;
(4) described precipitate is mixed with nucleic acid drug with aqueous solution dissolving is rear, centrifugalize obtains nanoparticle pharmaceutical composition;
Wherein, in step (1), described amphipathic nature polyalcohol is one or more in MPEG-PLA-glycolic, poly(ethylene oxide)-poly(propylene oxide) and Polyethylene Glycol and poly (l-lactic acid); Described hydrophilic small molecules medicine is one or more in amycin, mitoxantrone, daunorubicin and epirubicin;
In step (2), described first surface activating agent is one or more in polyvinyl alcohol, POLYSORBATE 80, span and dodecyl sodium sulfate; Described hydrophobic drug is one or more in paclitaxel, camptothecine, harringtonine and vinorelbine; Described cationic polymer be in poly-D-lysine, chitosan, Polyetherimide and polyamide-amide type dendrimer one or more;
In step (3), described second surface activating agent is one or more in polyvinyl alcohol, POLYSORBATE 80, span and dodecyl sodium sulfate;
In step (4), described nucleic acid drug is selected from DNA medicine and RNA medicine.
2. method according to claim 1, wherein, in step (1), the volume ratio of the aqueous solution of described the first solution and described hydrophilic small molecules medicine is 1:1-1:10; In described the first solution, with respect to first organic solvent of every milliliter, the content of described amphipathic nature polyalcohol is 5-100 milligram; In the aqueous solution of described hydrophilic small molecules medicine, with respect to the water of every milliliter, the content of described hydrophilic small molecules medicine is 1-100 milligram.
3. method according to claim 1 and 2, wherein, in step (1), described the first organic solvent is one or more in dichloromethane, dimethyl sulfoxide, oxolane, acetone, espeleton, dichloro-benzenes and methyl isopropyl ketone.
4. method according to claim 1, wherein, in step (1), the condition of described emulsifying comprises: the frequency of Ultrasonic Radiation is 20-25kHz, with respect to every milliliter of material of accepting Ultrasonic Radiation, power is 10-190W; The temperature of emulsifying is 1-99 DEG C, and the time of emulsifying is 1-30 minute.
5. method according to claim 1, wherein, in step (2), with respect to the first emulsion of 1 volume, the consumption of described first surface activating agent aqueous solution is 0.5-2 volume, the consumption of described the second solution is 0.1-1 volume, and the consumption of the aqueous solution of described cationic polymer is 0.1-1 volume; In described first surface activating agent aqueous solution, the content of described first surface activating agent is 0.6-6 % by weight; In described the second solution, with respect to second organic solvent of every milliliter, the content of described hydrophobic drug be 0.1 milligram to reach capacity in described the second organic solvent content of concentration of described hydrophobic drug; In the aqueous solution of described cationic polymer, with respect to the water of every milliliter, the content of described cationic polymer is for 1 milligram to reach capacity in the institute's aqueous solution content of concentration of described cationic polymer.
6. method according to claim 1 or 5, wherein, in step (2), the condition of described emulsifying comprises: the frequency of Ultrasonic Radiation is 20-25kHz, with respect to every milliliter of material of accepting Ultrasonic Radiation, power is 10-190W; The temperature of emulsifying is 1-99 DEG C, and the time of emulsifying is 1-30 minute.
7. method according to claim 1, wherein, in step (3), with respect to described second emulsion of 1 volume, the consumption of described second surface activating agent aqueous solution is 2-20 volume; In described second surface activating agent aqueous solution, the content of described second surface activating agent is 0.3-3 % by weight.
8. method according to claim 1, wherein, in step (4), with respect to the described precipitate of 1 weight portion, the consumption of water is 1-1000 weight portion, the consumption of described nucleic acid drug is 0.001-1 weight portion.
9. a nanoparticle pharmaceutical composition, this nanoparticle pharmaceutical composition is prepared by the method described in any one in claim 1-8, wherein, described hydrophobic drug is paclitaxel, described hydrophilic small molecules medicine is amycin, described nucleic acid drug is siRNA, described amphipathic nature polyalcohol is MPEG-PLA-ethanol copolymer, described cationic polymer is ε-poly-D-lysine, in described nanoparticle pharmaceutical composition, the weight ratio of described hydrophobic drug and described hydrophilic small molecules medicine is 1:10000-10000:1.
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