CN103520734B - A kind of based on albuminous nanoparticle, Preparation Method And The Use - Google Patents
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Abstract
The invention discloses a kind of based on albuminous nanoparticle, Preparation Method And The Use.Described nanoparticle comprises albumin molecule and positively charged macromole.Its preparation method utilizes positively charged macromole to carry out pretreatment to albumin, forms albumin nano particle by electrostatic interaction, and utilizes the compound containing sulfydryl to realize intermolecular cross-linking further, obtains stable albumin nano particle.Albumin nano particle preparation process of the present invention is without the need to an organic solvent, and obtained albumin nano particle footpath size is controlled, be evenly distributed, and can be used for drug loading.
Description
Technical field
The present invention relates to biological medicine new material technology field, particularly relate to a kind of based on albuminous nanoparticle, Preparation Method And The Use.
Background technology
Albumin is a class a large amount of albumen existed in blood plasma.In pharmaceutical carrier research, common albumin comprises human serum albumin, bovine serum albumin, oralbumin, can obtain respectively from human serum, Ox blood serum, Ovum Gallus domesticus album.Human serum albumin is plasma protein the abundantest in human body, and human serum albumin has multiple effect in vivo, as: the dissolubility 1) increasing long-chain fatty acid; 2) conjugated bilirubin; 3) be combined with multi-medicament, as medicines such as penicillins, sulfonamides, indoles, Benzodiazepineses.
Multi-medicament binding site is there is, to different types of medicine all energy payload in albumin molecule; It has the features such as water solublity is high, good stability, degradable simultaneously, and its biocompatibility is high, and be used for human body by U.S. food Drug Administration (FDA) approval, therefore albumin is the ideal carrier of drug conveying.Be used for drug conveying based on albuminous nanoparticle, the dissolubility of hydrophobic drug can be improved, and improve the dynamic metabolism of medicine.By controlling the particle diameter of nanoparticle, passive target effect can be improved, improving the picked-up of medicine in tumor site.Meanwhile, the amino that albumin exists, sulfydryl etc., can modify active targeting group further to improve targeting.
Prior art discloseder methods preparing albumin nano particle is divided into two parts usually, namely forms nanoparticle and stabilized nanoscale particle.The method forming albumin nano particle mainly contains desolventizing method and emulsion process.Desolventizing method (InternationalJournalofPharmaceutics, 2003,257,169 – 180) be add organic solvent in albumin aqueous solution, the dissolubility of albumin molecule is reduced, and produce and be separated and form nanoparticle, the volume fraction adding organic solvent need reach 40%.Emulsion process (InternationalJournalofPharmaceutics, 2007,340,163 – 172) uses Oleum Gossypii semen or cyclohexane give to be oil phase, has an effect with albuminous aqueous solution, produces nanoparticle.These processes forming albumin nano particle employ organic solvent, add the difficulty of post processing, and the organic solvent that simultaneously may remain can cause the adverse consequencess such as pharmaceutical properties instability, anaphylaxis.The common method stablizing albumin nano particle is cross-linking method.Glutaraldehydes (JournalofMicroencapsulation, 2001,18,825 – 829) etc. contain the cross-linking agent of aldehyde radical by the stable reaction albumin nano particle with amino.Owing to may produce toxic and side effects in vivo containing cross-link agent, such cross-linking agent is unsuitable for human body and uses.Have in report and employ glutathion or cysteine carries out pretreatment (Chinese patent application publication number CN102988996A) to albumin, stablize albumin nano particle by the effect of sulfhydryl and disulfide bond, but it forms nanoparticle also by adding organic solvent realization.
Summary of the invention
The object of the present invention is to provide a kind of based on albuminous nanoparticle, Preparation Method And The Use; Nanoparticle of the present invention has the controlled feature of even particle size distribution, size, its preparation process is not with an organic solvent, decrease the difficulty of post processing, the nanoparticle simultaneously formed also organic solvent-free remains, and avoids the adverse consequences such as pharmaceutical properties instability, anaphylaxis caused by organic solvent residual.
For reaching this object, the present invention by the following technical solutions:
First aspect, the invention provides a kind of based on albuminous nanoparticle, this nanoparticle comprises albumin molecule and positively charged macromole;
Preferably, described albumin molecule is any one or the two or more mixture in human serum albumin, bovine serum albumin, recombination human serum albumin, oralbumin;
Preferably, described positively charged macromole is any one or the two or more mixture in poly-D-lysine, spermine, poly-spermine, Protamine sulfates..
Second aspect, the invention provides the preparation method of nanoparticle described in first aspect, and described preparation method comprises the steps:
(1) mixed in aqueous with positively charged macromole by albumin molecule, the pH value regulating solution is 7.2-10.5;
(2) step (1) gained mixed liquor is reacted 1-60 minute, preferred 5-30 minute at 5-70 DEG C, preferably 10-60 DEG C, further preferred 20-40 DEG C;
(3) add the compound containing sulfydryl to step (2) gained mixed liquor, reaction more than 30 minutes at 5-70 DEG C, preferably 10-60 DEG C, further preferred 20-40 DEG C, forms stable albumin nano particle.
As preferably, in step (1), described albumin molecule concentration is in aqueous 1-200mg/ml, is preferably 10-100mg/ml, and described positively charged macromole concentration is in aqueous 1-200mg/ml, is preferably 10-100mg/ml;
Preferably, the pH value of solution is regulated by adding alkaline matter in aqueous solution; Further preferably, described alkaline matter is any one or the two or more mixture in sodium hydroxide, potassium hydroxide, sodium carbonate, potassium carbonate.
As preferably, containing any one or two or more mixture that the compound of sulfydryl is in glutathion, cysteine, dithiothreitol, DTT, mercaptoethanol described in step (3);
Preferably, the described final concentration of compound in mixed liquor containing sulfydryl is 0.001-100mM, is more preferably 0.1-10mM.
As preferably, above-mentioned preparation method also comprise step (3) gained is carried out dialysing containing the mixed liquor stablizing albumin nano particle, the step of processed;
Preferably, the temperature of described dialysis be 0 ~ 30 DEG C, preferably 4 ~ 28 DEG C further;
Preferably, described processed is high speed centrifugation, lyophilization, spraying dry or distilling under reduced pressure.
The third aspect, the invention provides a kind of based on albuminous nanoparticle, is obtained by preparation method described in second aspect present invention.
Fourth aspect, the invention provides nanoparticle described in first aspect present invention or the third aspect as the application of pharmaceutical carrier.
Preferably, described medicine is cancer therapy drug and/or optical dynamic therapy medicine;
Further preferably, described cancer therapy drug is any one or the two or more mixture in paclitaxel, Docetaxel, amycin, curcumin, cisplatin and analog thereof;
Further preferably, described optical dynamic therapy medicine is any one or the two or more mixture in temoporfin (Temoporfin), 5-ALA (5-Aminolevulinicacid), protoporphyrin IX (ProtoporphyrinIX), protoporphyrin IX disodium salt (ProtoporphyrinIXdisodium), Verteporfin (Verteporfin), single aspartyl chlorin (NPe6), tetraphenylporphyrin.
5th aspect, the invention provides a kind of preparation method of albumin nano particle of drug loading, comprises the steps:
(1) mixed in aqueous with positively charged macromole by albumin molecule, then add drug solution, the pH value regulating solution is 7.2-10.5, forms the suspension of the albumin nano particle of drug loading;
Preferably, described albumin molecule concentration is in aqueous 1-200mg/ml, is preferably 10-100mg/ml, and described positively charged macromole concentration is in aqueous 1-200mg/ml, is preferably 10-100mg/ml;
Preferably, described medicine is cancer therapy drug and/or optical dynamic therapy medicine; Further preferably, described cancer therapy drug is any one or the two or more mixture in paclitaxel, Docetaxel, amycin, curcumin, cisplatin and analog thereof; Further preferably, described optical dynamic therapy medicine is any one or the two or more mixture in temoporfin, 5-ALA, protoporphyrin IX, protoporphyrin IX disodium salt, Verteporfin, single aspartyl chlorin, tetraphenylporphyrin;
Preferably, the solvent of described drug solution is any one or the two or more mixture in water, ethanol, acetone, dimethyl sulfoxide, oxolane;
(2) step (1) gained suspension reacts 1-60 minute, preferred 5-30 minute at 5-70 DEG C, preferably 10-60 DEG C, further preferred 20-40 DEG C;
(3) add the compound containing sulfydryl to step (2) gained suspension, reaction more than 30 minutes at 5-70 DEG C, preferably 10-60 DEG C, further preferred 20-40 DEG C, obtains the sub-suspension of albumin nano granular of stable drug loading;
Preferably, the described compound containing sulfydryl is any one or the two or more mixture in glutathion, cysteine, dithiothreitol, DTT, mercaptoethanol;
Preferably, the described final concentration of compound in reactant liquor containing sulfydryl is 0.001-100mM, is more preferably 0.1-10mM;
(4) the sub-suspension of albumin nano granular stable for step (3) gained is carried out dialyse, processed, obtain the albumin nano particle of drug loading;
Preferably, the temperature of described dialysis is 0 ~ 30 DEG C, more preferably 4 ~ 28 DEG C;
Preferably, described processed is high speed centrifugation, lyophilization, spraying dry or distilling under reduced pressure.
6th aspect, the invention provides a kind of albumin nano particle of drug loading, is obtained by preparation method described in fifth aspect present invention.
Preparation method based on albuminous nanoparticle of the present invention utilizes positively charged macromole to carry out pretreatment to albumin, albumin nano particle is formed by electrostatic interaction, and utilize the compound containing sulfydryl to realize intermolecular cross-linking further, obtain stable albumin nano particle; Above-mentioned preparation method not with an organic solvent, decreases the difficulty of post processing, and the nanoparticle simultaneously formed also organic solvent-free remains, and avoids the adverse consequences such as pharmaceutical properties instability, anaphylaxis caused by organic solvent residual.Above-mentioned preparation method gained albumin nano particle footpath is evenly distributed, size is controlled, and mean diameter can regulate in 10 ~ 5000nm.
Accompanying drawing explanation
Fig. 1 is the scanning electron microscope diagram sheet of embodiment 1 gained albumin nano particle.
Fig. 2 is the scanning electron microscope diagram sheet of embodiment 2 gained albumin nano particle.
Fig. 3 is the grain size distribution that embodiment 3 gained is loaded with the albumin nano particle of cisplatin.
Fig. 4 is the laser co-focusing picture that embodiment 4 gained is loaded with the albumin nano particle of amycin.
Fig. 5 is that embodiment 6 gained is loaded with the albumin nano particle of paclitaxel and the particle diameter of blank nanoparticle and potential image; A is the particle diameter of blank nanosphere, and B is the particle diameter of the nanoparticle carrying paclitaxel, and C is the current potential of blank nanoparticle, and D is the particle diameter of the nanoparticle carrying paclitaxel.
Fig. 6 is the albumin nano particle uv-visible absorption spectra that embodiment 9 gained is loaded with protoporphyrin IX disodium salt.
Detailed description of the invention
Describe the present invention by the following examples, but the present invention is not limited in following embodiment.
Embodiment 1
Configuration contains the 1ml aqueous solution of 1mg human serum albumin and 0.6mg poly-D-lysine, sodium hydroxide is added after mixing, adjust ph to 8.5, leaves standstill the 0.1M dithiothreitol, DTT solution adding 10 μ L after 5 minutes, the albumin nano particle that left at room temperature over night must be stable.
The scanning electron microscope diagram sheet of gained nanoparticle is as accompanying drawing 1.
Embodiment 2
Configuration contains the 1ml aqueous solution of 1mg human serum albumin and 1mg poly-D-lysine, sodium hydroxide is added after mixing, adjust ph to 9.5, leave standstill the 0.1M dithiothreitol, DTT solution adding 10 μ L after 60 minutes, room temperature places albumin nano particle that 8 hours must be stable.
The scanning electron microscope diagram sheet of gained nanoparticle is as accompanying drawing 2.
Embodiment 3
Getting 10mg bovine serum albumin is dissolved in 1ml water, and add 10mg poly-D-lysine, the 1mg/ml aqueous solution of 100 μ L cisplatin is added after mixing, add potassium hydroxide, adjust ph to 8.2, the suspension obtained reacts 30 minutes under room temperature, then add wherein 10 μ L 1M cysteine and in left at room temperature 30 minutes, by suspension high speed centrifugation, obtain albumin nano particle powder.
Gained contains the grain size distribution of the nanoparticle of cisplatin as accompanying drawing 3.
Embodiment 4
Getting 100mg oralbumin is dissolved in 1ml water, and add 50mg and gather spermine, the 1mg/ml aqueous solution of 200 μ L amycin is added after mixing, add sodium carbonate, adjust ph to 10.5, the suspension obtained reacts 30 minutes at 60 DEG C, then adds the 1M cysteine of 100 μ L wherein and react 2 hours at 60 DEG C, suspension to be dialysed postlyophilization with water at 4 DEG C, obtains the albumin nano particle powder being loaded with amycin.
Amycin has fluorescence, and other compounds in preparation process all do not have fluorescence, and the fluorescent characteristic of nanoparticle can determine that amycin is enclosed in nanoparticle, and gained contains the Laser Scanning Confocal Microscope picture of the nanoparticle of amycin as accompanying drawing 4.
Embodiment 5
Getting 10mg recombination human serum albumin is dissolved in 1ml water, and add 10mg and gather spermine, the 1mg/ml aqueous solution of 100 μ L curcumins is added after mixing, add potassium carbonate, adjust ph to 9.0, the suspension obtained reacts 30 minutes at 20 DEG C, then adds the 1M mercaptoethanol of 10 μ L wherein and react 8 hours at 20 DEG C, carry out spraying dry after being dialysed with water at 30 DEG C by suspension, obtain albumin nano particle powder.
Embodiment 6
Getting 1mg bovine serum albumin is dissolved in 1ml water, and add 0.6mg poly-D-lysine, add the 10mg/ml alcoholic solution of 10 μ L paclitaxels after mixing, add potassium hydroxide after mixing, adjust ph to 9.0, the suspension obtained leaves standstill 5 minutes at 20 DEG C, add the 0.1M dithiothreitol, DTT of 10 μ L more wherein, at 20 DEG C, leave standstill 8 hours, after suspension and water being dialysed, carry out high speed centrifugation, collecting precipitation, drying, obtain the albumin nano particle powder being surrounded by paclitaxel.
Gained contained the nanoparticle of paclitaxel and do not compare containing the blank nanoparticle of paclitaxel, both particle diameters and current potential all do not have significant difference, illustrate that the process of drug loading does not affect the character of nanoparticle.Containing the particle diameter of paclitaxel and blank nanoparticle and potential image as accompanying drawing 5, wherein A is the particle diameter of blank nanosphere, and B is the particle diameter of the nanoparticle carrying paclitaxel, and C is the current potential of blank nanoparticle, and D is the particle diameter of the nanoparticle carrying paclitaxel.
Embodiment 7
Getting 200mg bovine serum albumin is dissolved in 1ml water, and add 200mg and gather spermine, the 1mg/ml alcoholic solution of 100 μ L temoporfins is added after mixing, add potassium hydroxide, adjust ph to 9.0, the suspension obtained reacts 30 minutes at 20 DEG C, then adds the 1M dithiothreitol, DTT of 100 μ L wherein and leave standstill 8 hours at 20 DEG C, by suspension high speed centrifugation, obtain albumin nano particle powder.
Embodiment 8
Getting 10mg bovine serum albumin is dissolved in 1ml water, and add 10mg poly-D-lysine, the 10mg/ml aqueous solution of the 5-ALA of 200 μ L is added after mixing, add potassium hydroxide, adjust ph to 9.0, the suspension obtained reacts 30 minutes at 20 DEG C, then adds the 1M dithiothreitol, DTT of 10 μ L wherein and react 2 hours at 20 DEG C, by suspension high speed centrifugation, obtain albumin nano particle powder.
Embodiment 9
Getting 10mg bovine serum albumin is dissolved in 1ml water, and add 10mg poly-D-lysine, the 0.5mg/ml aqueous solution of 200 μ L protoporphyrin IX disodium salts is added after mixing, add potassium hydroxide, adjust ph to 9.5, the suspension obtained reacts 30 minutes at 20 DEG C, then adds the 1M dithiothreitol, DTT of 10 μ L wherein and react 2 hours at 20 DEG C, by suspension high speed centrifugation, obtain albumin nano particle powder.
By soluble in water for the albumin nano particle obtained, test ultraviolet-visible absorption spectroscopy, occur the characteristic absorption peak of protoporphyrin IX disodium salt at 390nm place, illustrate that protoporphyrin IX disodium salt is loaded in nanoparticle, the ultraviolet-visible absorption spectra after normalization is shown in accompanying drawing 6.
Embodiment 10
Getting 10mg bovine serum albumin is dissolved in 1ml water, and add 10mg Protamine sulfates., the 5mg/ml tetrahydrofuran solution of 100 μ L Verteporfins is added after mixing, add potassium hydroxide, adjust ph to 9.5, the suspension obtained reacts 30 minutes at 20 DEG C, then adds the 1M dithiothreitol, DTT of 10 μ L wherein and react 2 hours at 20 DEG C, suspension to be dialysed postlyophilization with water at 4 DEG C, obtains albumin nano particle powder.
Embodiment 11
Getting 10mg bovine serum albumin is dissolved in 1ml water; and add 10mg and gather spermine; the 5mg/ml aqueous solution of the mono-aspartyl chlorin of 100 μ L is added after mixing; add potassium hydroxide; adjust ph to 9.0, the suspension obtained reacts 30 minutes at 20 DEG C, then adds the 1M dithiothreitol, DTT of 10 μ L wherein and react 2 hours at 20 DEG C; by suspension high speed centrifugation, obtain albumin nano particle powder.
Embodiment 12
Getting 10mg bovine serum albumin is dissolved in 1ml water, and add 10mg and gather spermine, the DMSO solution of the 5mg/ml of 10 μ L tetraphenylporphyrins is added after mixing, add potassium hydroxide, adjust ph to 9.0, the suspension obtained reacts 30 minutes at 20 DEG C, then adds the 1M glutathion of 10 μ L wherein and react 2 hours at 20 DEG C, by suspension high speed centrifugation, obtain albumin nano particle powder.
Embodiment 13
Getting 1mg bovine serum albumin is dissolved in 1ml water, and add 1mg and gather spermine, the acetone soln of the 1mg/ml of 10 μ L protoporphyrin IXs is added after mixing, add potassium hydroxide, adjust ph to 9.0, the suspension obtained reacts 30 minutes at 20 DEG C, then adds the 0.1M glutathion of 10 μ L wherein and leave standstill 8 hours at 20 DEG C, by suspension high speed centrifugation, after drying, obtain the albumin nano particle powder containing protoporphyrin IX.
Applicant states, the present invention illustrates product of the present invention and detailed preparation method by above-described embodiment, but the present invention is not limited to the said goods and detailed preparation method, namely do not mean that the present invention must rely on the said goods and detailed preparation method could be implemented.Person of ordinary skill in the field should understand, any improvement in the present invention, to equivalence replacement and the interpolation of auxiliary element, the concrete way choice etc. of each raw material of product of the present invention, all drops within protection scope of the present invention and open scope.
Claims (46)
1. based on an albuminous nanoparticle, it is characterized in that, comprise albumin molecule and positively charged macromole;
Described albumin molecule is any one or two or more mixture in human serum albumin, bovine serum albumin, recombination human serum albumin, oralbumin;
Described positively charged macromole is any one or two or more mixture in poly-D-lysine, spermine, poly-spermine, Protamine sulfates.;
Describedly to be made up of the preparation method comprising the steps based on albuminous nanoparticle:
(1) mixed in aqueous with positively charged macromole by albumin molecule, the pH value regulating solution is 7.2-10.5;
(2) step (1) gained mixed liquor is reacted 1-60 minute at 5-70 DEG C;
(3) add the compound containing sulfydryl to step (2) gained mixed liquor, react more than 30 minutes at 5-70 DEG C, form stable albumin nano particle.
2. according to claim 1ly it is characterized in that based on albuminous nanoparticle, in described preparation method, the reaction temperature of step (2) is 10-60 DEG C.
3. according to claim 1ly it is characterized in that based on albuminous nanoparticle, in described preparation method, the reaction temperature of step (2) is 20-40 DEG C.
4. according to claim 1ly it is characterized in that based on albuminous nanoparticle, in described preparation method, the response time of step (2) is 5-30 minute.
5. according to claim 1ly it is characterized in that based on albuminous nanoparticle, in described preparation method, the reaction temperature of step (3) is 10-60 DEG C.
6. according to claim 1ly it is characterized in that based on albuminous nanoparticle, in described preparation method, the reaction temperature of step (3) is 20-40 DEG C.
7. described in any one of claim 1-6 based on the preparation method of albuminous nanoparticle, it is characterized in that, comprise the steps:
(1) mixed in aqueous with positively charged macromole by albumin molecule, the pH value regulating solution is 7.2-10.5;
(2) step (1) gained mixed liquor is reacted 1-60 minute at 5-70 DEG C;
(3) add the compound containing sulfydryl to step (2) gained mixed liquor, react more than 30 minutes at 5-70 DEG C, form stable albumin nano particle.
8. preparation method according to claim 7, is characterized in that, in step (1), described albumin molecule concentration is in aqueous 1-200mg/ml, and described positively charged macromole concentration is in aqueous 1-200mg/ml.
9. preparation method according to claim 8, is characterized in that, in step (1), described albumin molecule concentration is in aqueous 10-100mg/ml.
10. preparation method according to claim 8, is characterized in that, in step (1), described positively charged macromole concentration is in aqueous 10-100mg/ml.
11. preparation methoies according to claim 8, is characterized in that, in step (1), regulate the pH value of solution by adding alkaline matter in aqueous solution.
12. preparation methoies according to claim 11, is characterized in that, described alkaline matter is any one or two or more mixture in sodium hydroxide, potassium hydroxide, sodium carbonate, potassium carbonate.
13. preparation methoies according to claim 7, is characterized in that, the reaction temperature of step (2) is 10-60 DEG C.
14. preparation methoies according to claim 7, is characterized in that, the reaction temperature of step (2) is 20-40 DEG C.
15. preparation methoies according to claim 7, is characterized in that, the response time of step (2) is 5-30 minute.
16. preparation methoies according to claim 7, is characterized in that, the reaction temperature of step (3) is 10-60 DEG C.
17. preparation methoies according to claim 7, is characterized in that, the reaction temperature of step (3) is 20-40 DEG C.
18. preparation methoies according to claim 7, is characterized in that, in step (3), the described compound containing sulfydryl is any one or two or more mixture in glutathion, cysteine, dithiothreitol, DTT, mercaptoethanol.
19. preparation methoies according to claim 18, is characterized in that, the described final concentration of compound in mixed liquor containing sulfydryl is 0.001-100mM.
20. preparation methoies according to claim 19, is characterized in that, the described final concentration of compound in mixed liquor containing sulfydryl is 0.1-10mM.
21. preparation methoies according to any one of claim 7-20, is characterized in that, also comprise step (3) gained is carried out dialysing containing the mixed liquor stablizing albumin nano particle, the step of processed.
22. preparation methoies according to claim 21, is characterized in that, the temperature of described dialysis is 0 ~ 30 DEG C.
23. preparation methoies according to claim 22, is characterized in that, the temperature of described dialysis is 4 ~ 28 DEG C.
24. preparation methoies according to claim 21, is characterized in that, described processed is high speed centrifugation, lyophilization, spraying dry or distilling under reduced pressure.
25. 1 kinds, based on albuminous nanoparticle, are obtained by preparation method described in any one of claim 2-24.
26. as described in any one of claim 1-6 or claim 25 nanoparticle as the application of pharmaceutical carrier in useful in preparing drug formulations.
27. application according to claim 26, is characterized in that, described medicine is cancer therapy drug and/or optical dynamic therapy medicine.
28. application according to claim 27, is characterized in that, described cancer therapy drug is any one or two or more mixture in paclitaxel, Docetaxel, amycin, curcumin, cisplatin and analog thereof; Described optical dynamic therapy medicine is any one or two or more mixture in temoporfin, 5-ALA, protoporphyrin IX, protoporphyrin IX disodium salt, Verteporfin, single aspartyl chlorin, tetraphenylporphyrin.
The preparation method of the albumin nano particle of 29. 1 kinds of drug loading, is characterized in that, comprise the steps:
(1) mixed in aqueous with positively charged macromole by albumin molecule, then add drug solution, the pH value regulating solution is 7.2-10.5, forms the suspension of the albumin nano particle of drug loading;
Wherein, described albumin molecule is any one or the two or more mixture in human serum albumin, bovine serum albumin, recombination human serum albumin, oralbumin; Described positively charged macromole is any one or two or more mixture in poly-D-lysine, spermine, poly-spermine, Protamine sulfates.;
(2) step (1) gained suspension reacts 1-60 minute at 5-70 DEG C;
(3) add the compound containing sulfydryl to step (2) gained suspension, react more than 30 minutes at 5-70 DEG C, obtain the sub-suspension of albumin nano granular of stable drug loading;
(4) the sub-suspension of albumin nano granular stable for step (3) gained is carried out dialyse, processed, obtain the albumin nano particle of drug loading.
30. preparation methoies according to claim 29, it is characterized in that, in step (1), described albumin molecule concentration is in aqueous 1-200mg/ml, described positively charged macromole concentration is in aqueous 1-200mg/ml, and described medicine is cancer therapy drug and/or optical dynamic therapy medicine.
31. preparation methoies according to claim 30, is characterized in that, described albumin molecule concentration is in aqueous 10-100mg/ml.
32. preparation methoies according to claim 30, is characterized in that, described positively charged macromole concentration is in aqueous 10-100mg/ml.
33. preparation methoies according to claim 30, is characterized in that, described cancer therapy drug is any one or two or more mixture in paclitaxel, Docetaxel, amycin, curcumin, cisplatin and analog thereof; Described optical dynamic therapy medicine is any one or two or more mixture in temoporfin, 5-ALA, protoporphyrin IX, protoporphyrin IX disodium salt, Verteporfin, single aspartyl chlorin, tetraphenylporphyrin.
34. preparation methoies according to claim 33, is characterized in that, the solvent of described drug solution is any one or two or more mixture in water, ethanol, acetone, dimethyl sulfoxide, oxolane.
35. preparation methoies according to claim 29, is characterized in that, the reaction temperature of step (2) is 10-60 DEG C.
36. preparation methoies according to claim 29, is characterized in that, the reaction temperature of step (2) is 20-40 DEG C.
37. preparation methoies according to claim 29, is characterized in that, the response time of step (2) is 5-30 minute.
38. preparation methoies according to claim 29, is characterized in that, the reaction temperature of step (3) is 10-60 DEG C.
39. preparation methoies according to claim 29, is characterized in that, the reaction temperature of step (3) is 20-40 DEG C.
40. preparation methoies according to claim 29, is characterized in that, in step (3), the described compound containing sulfydryl is any one or two or more mixture in glutathion, cysteine, dithiothreitol, DTT, mercaptoethanol.
41. preparation methoies according to claim 40, is characterized in that, the described final concentration of compound in reactant liquor containing sulfydryl is 0.001-100mM.
42. preparation methoies according to claim 41, is characterized in that, the described final concentration of compound in reactant liquor containing sulfydryl is 0.1-10mM.
43. preparation methoies according to claim 29, is characterized in that, in step (4), the temperature of described dialysis is 0 ~ 30 DEG C.
44. preparation methoies according to claim 43, is characterized in that, in step (4), the temperature of described dialysis is 4 ~ 28 DEG C.
45. preparation methoies according to claim 29, is characterized in that, in step (4), described processed is high speed centrifugation, lyophilization, spraying dry or distilling under reduced pressure.
The albumin nano particle of 46. 1 kinds of drug loading, is obtained by preparation method described in any one of claim 29-45.
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| CN117442702A (en) * | 2022-01-26 | 2024-01-26 | 陕西师范大学 | Antibacterial liquid with controllable-size protein-based nanoparticles |
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| CN101138636A (en) * | 2006-09-05 | 2008-03-12 | 中国科学院上海药物研究所 | Gene medicine conveying system and method of preparing the same |
| CN102988996A (en) * | 2012-12-19 | 2013-03-27 | 清华大学 | Method for preparing stable albumin nanoparticle |
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| CN101138636A (en) * | 2006-09-05 | 2008-03-12 | 中国科学院上海药物研究所 | Gene medicine conveying system and method of preparing the same |
| CN102988996A (en) * | 2012-12-19 | 2013-03-27 | 清华大学 | Method for preparing stable albumin nanoparticle |
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