CN104644614A - Supported hemoglobin composition and preparation method thereof - Google Patents
Supported hemoglobin composition and preparation method thereof Download PDFInfo
- Publication number
- CN104644614A CN104644614A CN201510069797.5A CN201510069797A CN104644614A CN 104644614 A CN104644614 A CN 104644614A CN 201510069797 A CN201510069797 A CN 201510069797A CN 104644614 A CN104644614 A CN 104644614A
- Authority
- CN
- China
- Prior art keywords
- hemoglobin
- aqueous solution
- peg
- preparation
- ultrasonic
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 108010054147 Hemoglobins Proteins 0.000 title claims abstract description 128
- 102000001554 Hemoglobins Human genes 0.000 title claims abstract description 128
- 239000000203 mixture Substances 0.000 title claims abstract description 60
- 238000002360 preparation method Methods 0.000 title claims abstract description 32
- 238000000034 method Methods 0.000 claims abstract description 27
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 18
- 239000000839 emulsion Substances 0.000 claims abstract description 14
- 239000003960 organic solvent Substances 0.000 claims abstract description 14
- 229920001606 poly(lactic acid-co-glycolic acid) Polymers 0.000 claims abstract description 14
- 239000004094 surface-active agent Substances 0.000 claims abstract description 13
- 238000002156 mixing Methods 0.000 claims abstract description 6
- 229920001400 block copolymer Polymers 0.000 claims description 60
- DNIAPMSPPWPWGF-UHFFFAOYSA-N Propylene glycol Chemical compound CC(O)CO DNIAPMSPPWPWGF-UHFFFAOYSA-N 0.000 claims description 48
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 45
- 239000007864 aqueous solution Substances 0.000 claims description 39
- 239000000243 solution Substances 0.000 claims description 39
- 239000004372 Polyvinyl alcohol Substances 0.000 claims description 24
- 229920002451 polyvinyl alcohol Polymers 0.000 claims description 24
- 239000005457 ice water Substances 0.000 claims description 20
- 239000004721 Polyphenylene oxide Substances 0.000 claims description 16
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 claims description 16
- 229920000570 polyether Polymers 0.000 claims description 16
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 15
- 229960004275 glycolic acid Drugs 0.000 claims description 13
- AEMRFAOFKBGASW-UHFFFAOYSA-N Glycolic acid Chemical compound OCC(O)=O AEMRFAOFKBGASW-UHFFFAOYSA-N 0.000 claims description 12
- 210000003022 colostrum Anatomy 0.000 claims description 10
- 235000021277 colostrum Nutrition 0.000 claims description 10
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 8
- 238000012661 block copolymerization Methods 0.000 claims description 8
- 239000004310 lactic acid Substances 0.000 claims description 8
- 235000014655 lactic acid Nutrition 0.000 claims description 8
- 229920001223 polyethylene glycol Polymers 0.000 claims description 7
- 239000000178 monomer Substances 0.000 claims description 6
- 239000002202 Polyethylene glycol Substances 0.000 claims description 5
- 238000006116 polymerization reaction Methods 0.000 claims description 3
- 238000005119 centrifugation Methods 0.000 claims description 2
- 239000000758 substrate Substances 0.000 claims description 2
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 abstract description 17
- 210000004369 blood Anatomy 0.000 abstract description 17
- 239000008280 blood Substances 0.000 abstract description 17
- 229910052760 oxygen Inorganic materials 0.000 abstract description 12
- 239000001301 oxygen Substances 0.000 abstract description 12
- 230000002776 aggregation Effects 0.000 abstract description 6
- 238000004220 aggregation Methods 0.000 abstract description 6
- 210000003743 erythrocyte Anatomy 0.000 abstract description 6
- 230000008901 benefit Effects 0.000 abstract description 5
- 239000004698 Polyethylene Substances 0.000 abstract description 4
- -1 polyethylene Polymers 0.000 abstract description 4
- 229920000573 polyethylene Polymers 0.000 abstract description 4
- 239000002994 raw material Substances 0.000 abstract description 3
- 239000012876 carrier material Substances 0.000 abstract description 2
- 238000009210 therapy by ultrasound Methods 0.000 abstract 2
- 150000001875 compounds Chemical class 0.000 abstract 1
- 238000004945 emulsification Methods 0.000 abstract 1
- 231100000956 nontoxicity Toxicity 0.000 abstract 1
- 239000002105 nanoparticle Substances 0.000 description 56
- 210000004027 cell Anatomy 0.000 description 18
- 239000002245 particle Substances 0.000 description 18
- 230000009514 concussion Effects 0.000 description 12
- 239000003633 blood substitute Substances 0.000 description 8
- 239000000463 material Substances 0.000 description 8
- 238000009826 distribution Methods 0.000 description 7
- 230000000694 effects Effects 0.000 description 6
- 239000012530 fluid Substances 0.000 description 6
- 239000006228 supernatant Substances 0.000 description 6
- 238000012360 testing method Methods 0.000 description 6
- 239000002502 liposome Substances 0.000 description 5
- 230000008859 change Effects 0.000 description 4
- 238000006243 chemical reaction Methods 0.000 description 4
- 238000001727 in vivo Methods 0.000 description 4
- 210000002540 macrophage Anatomy 0.000 description 4
- 230000000242 pagocytic effect Effects 0.000 description 4
- 230000008569 process Effects 0.000 description 4
- 206010029155 Nephropathy toxic Diseases 0.000 description 3
- 206010047139 Vasoconstriction Diseases 0.000 description 3
- 230000005540 biological transmission Effects 0.000 description 3
- 238000007385 chemical modification Methods 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 239000003814 drug Substances 0.000 description 3
- 239000003094 microcapsule Substances 0.000 description 3
- 210000000865 mononuclear phagocyte system Anatomy 0.000 description 3
- 230000007694 nephrotoxicity Effects 0.000 description 3
- 231100000417 nephrotoxicity Toxicity 0.000 description 3
- 210000001519 tissue Anatomy 0.000 description 3
- 230000025033 vasoconstriction Effects 0.000 description 3
- RKDVKSZUMVYZHH-UHFFFAOYSA-N 1,4-dioxane-2,5-dione Chemical compound O=C1COC(=O)CO1 RKDVKSZUMVYZHH-UHFFFAOYSA-N 0.000 description 2
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 2
- 206010018910 Haemolysis Diseases 0.000 description 2
- 239000012736 aqueous medium Substances 0.000 description 2
- 230000023555 blood coagulation Effects 0.000 description 2
- 238000002983 circular dichroism Methods 0.000 description 2
- 239000006185 dispersion Substances 0.000 description 2
- 230000008588 hemolysis Effects 0.000 description 2
- 230000002209 hydrophobic effect Effects 0.000 description 2
- JJTUDXZGHPGLLC-UHFFFAOYSA-N lactide Chemical compound CC1OC(=O)C(C)OC1=O JJTUDXZGHPGLLC-UHFFFAOYSA-N 0.000 description 2
- 210000002381 plasma Anatomy 0.000 description 2
- 229920000747 poly(lactic acid) Polymers 0.000 description 2
- 239000004626 polylactic acid Substances 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 238000001338 self-assembly Methods 0.000 description 2
- 241001436679 Adama Species 0.000 description 1
- 101710088194 Dehydrogenase Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 241000662429 Fenerbahce Species 0.000 description 1
- 206010020772 Hypertension Diseases 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 108010061951 Methemoglobin Proteins 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 244000309464 bull Species 0.000 description 1
- 230000009172 bursting Effects 0.000 description 1
- 239000000969 carrier Substances 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 238000001142 circular dichroism spectrum Methods 0.000 description 1
- 238000013270 controlled release Methods 0.000 description 1
- 229920001577 copolymer Polymers 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003937 drug carrier Substances 0.000 description 1
- 230000000857 drug effect Effects 0.000 description 1
- 230000037149 energy metabolism Effects 0.000 description 1
- 210000003617 erythrocyte membrane Anatomy 0.000 description 1
- 230000003203 everyday effect Effects 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 230000001631 hypertensive effect Effects 0.000 description 1
- 238000002513 implantation Methods 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 238000001802 infusion Methods 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 125000000956 methoxy group Chemical group [H]C([H])([H])O* 0.000 description 1
- 230000004899 motility Effects 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 238000011275 oncology therapy Methods 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 230000003204 osmotic effect Effects 0.000 description 1
- 239000005022 packaging material Substances 0.000 description 1
- 238000005502 peroxidation Methods 0.000 description 1
- 239000000546 pharmaceutical excipient Substances 0.000 description 1
- 229920005643 polyisobutyl cyanoacrylate Polymers 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 238000013268 sustained release Methods 0.000 description 1
- 239000012730 sustained-release form Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 230000008685 targeting Effects 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000004102 tricarboxylic acid cycle Effects 0.000 description 1
Landscapes
- Medicinal Preparation (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The invention relates to a supported hemoglobin composition and a preparation method thereof. The preparation method comprises the following steps: (1) dissolving a carrier polyethylene glycol-poly(lactide-co-glycolide) into an organic solvent, mixing with a water solution of hemoglobin, and carrying out ultrasonic treatment to form primary emulsion; (2) adding a water solution of a surfactant to the primary emulsion obtained in the step (1), mixing and carrying out ultrasonic treatment to form a compound emulsion; and (3) removing the organic solvent, and centrifuging to obtain the supported hemoglobin composition. The supported hemoglobin composition is prepared by a double emulsion method; the method is simple and feasible; the prepared supported hemoglobin composition has oxygen carrying and releasing functions, has rheological characteristics similar to whole blood, is high in stability and good in blood compatibility, and does not cause erythrocyte aggregation; and the carrier material provided by the invention is prepared from abundant raw material, has the advantages of low cost, biocompatibility, biological affinity and no toxicity and is prone to be chemically modified.
Description
Technical field
The invention belongs to field of medicinal compositions, particularly relate to a kind of support type hemoglobin compositions and preparation method thereof.
Background technology
Blood transfusion has very important effect in surgical medicine and treatment of war wound, and along with the variation of human lives and complicated, the supply source of blood and safety issue more and more become the important content of economic society livelihood issues.Because blood substitute has the advantage of " versatility is good, safety is high, long shelf-life ", make the research and development of blood substitute significant all the time and receive much concern.Wherein, based on the carrier of oxygen (the hemoglobin-based oxygen carriers of hemoglobin, HBOCs) be the study hotspot of blood substitute field, be divided into conjugate and class cellularity load hemoglobin three class of the hemoglobin of chemical modification, hemoglobin and various enzyme body.Wherein class cellularity load hemoglobin can avoid hemoglobin directly with surrounding tissue and other composition direct effects of blood, prevent vasoconstriction, avoid nephrotoxicity, raising circulation time in vivo.This kind can be divided into loading liposomes hemoglobin and macromolecular material load hemoglobin according to material difference.Liposome peroxidation can occur in loading liposomes hemoglobin process, reductase system can not pass liposome membrane, makes hemoglobin oxidizable.Compared with liposome, biodegradable macromolecular material has following several advantage in load hemoglobin: mechanical strength is high, preparation process is easy to control particle size and grain thickness, circulation time in vivo and stability are stronger, and the motility of its design is more easily connected longer PEG chain with intellectuality.
Biodegradable Poly(D,L-lactide-co-glycolide [poly (D, L-lactide-co-glycolid), PLGA] be high-molecular copolymer by lactide (LA) and Acetic acid, hydroxy-, bimol. cyclic ester (GA) two kinds of monomer polymerizations, it is the synthesis high-molecular biologic degradable material being widely used in medicine carrying controlled release system at present, its degradation mechanism is be hydrolyzed defat in vivo to generate lactic acid monomer, and acetone acid is generated under lactic acid dehydrogenase effect, participate in tricarboxylic acid cycle in body as one of energy metabolism material.PLGA has good biocompatibility, does not have obvious inflammatory reaction, immunoreation and cell-cytotoxic reaction.PLGA by U.S. FDA approval formally entered American Pharmacopeia as pharmaceutic adjuvant, in recent years also for sustained-release drug carrier and other human bodies implantation device materials.Be that the nanoparticle that carrier is made has slow-releasing with PLGA, administration number of times and dose can be reduced, strengthen organ targeting and drug effect, be therefore widely used.Polyethylene Glycol (PEG) good hydrophilic property, minimizing protein attachment, cover nanoparticle surface, macrophage phagocytic in mononuclear phagocyte system (mononuclear phagocyte system, MPS) can be reduced, the circulation time in extension body.
Chang etc. once used PLA, PLGA and Polyisobutyl cyanoacrylate (polyisobutycyanoacrylate) mixture as a kind of packaging material parcel hemoglobin in 1997, particle size distribution is from 70nm to 1110nm, but its safety and effectiveness have no report.
CN1419936A discloses a kind of blood substitute of hemoglobin microcapsule and preparation method thereof, this invention contains one or more polylactic acid mono methoxy polyethylene glycol copolymer and hemoglobin by microcapsule prepared by double emulsion, preparation method technique is simple, easy amplification, and consuming time short, but this particle diameter inventing the hemoglobin microcapsule prepared is several micron, particle diameter is larger, easily by macrophage phagocytic in mononuclear phagocyte system, circulation time is in vivo shorter.
CN102861322A discloses a kind of hemoglobin O_2 carrier and preparation method thereof, this hemoglobin O_2 carrier comprises hemoglobin, hoptoglobin and polyethylene glycol-ester nanoparticles, hemoglobin is connected to polyethylene glycol-ester nanoparticles surface by hoptoglobin, hemoglobin O_2 carrier of the present invention to cell without overt toxicity, after infusion in blood plasma without obvious free hemoglobin, without hyper tensive reactions, the defect that the side effect of traditional hemoglobin O_2 carrier is strong can be overcome, have a good application prospect and economic benefit.But this invention is when preparing this hemoglobin O_2 carrier, needs to carry out chemical modification to polymeric material, and by hoptoglobin, hemoglobin is bonded to polyethylene glycol-ester nanoparticles surface, technics comparing is complicated.
Therefore, this area is expected to utilize better simply method to develop a kind ofly to have compared with small particle diameter and the hemoglobin-based oxygen carrier safely and effectively that hemoglobin can be protected not destroyed.
Summary of the invention
For the deficiencies in the prior art, the object of the present invention is to provide a kind of support type hemoglobin compositions and preparation method thereof.
For reaching this goal of the invention, the present invention by the following technical solutions:
On the one hand, the invention provides a kind of preparation method of support type hemoglobin compositions, said method comprising the steps of:
(1) carrier PEG-PLGA block copolymer is dissolved in organic solvent, then by the aqueous solution of itself and hemoglobin, ultrasonic, forms colostrum;
(2) just the aqueous solution of surfactant is added in Ruzhong to step (1) gained, mixing, ultrasonic, form emulsion;
(3) organic solvent is removed, centrifugal, obtain support type hemoglobin compositions.
In the preparation method of support type hemoglobin compositions of the present invention, the structural formula of step (1) described carrier PEG-PLGA block copolymer (PEG-PLGA) is as follows:
Preferably, in described PEG-PLGA block copolymer, the weight average molecular weight of Polyethylene Glycol segment is 1000-5000Da, such as 1000Da, 2000Da, 3000Da, 4000Da or 5000Da, the weight average molecular weight of polylactic-co-glycolic acid segment is 4500-45000Da, such as 4500Da, 5000Da, 7500Da, 9000Da, 10000Da, 20000Da, 30000Da, 40000Da or 45000Da; Preferably, described polylactic-co-glycolic acid segment is formed by lactic acid and hydroxyacetic acid two kinds of monomer polymerizations, the mol ratio of wherein said lactic acid and hydroxyacetic acid two kinds of monomers is 75:25 or 50:50, and namely described polylactic-co-glycolic acid segment is made up of 75% lactic acid and 25% hydroxyacetic acid or is made up of (being PLGA75/25 or PLGA50/50) 50% lactic acid and 50% hydroxyacetic acid.
In the preparation method of support type hemoglobin compositions of the present invention, step (1) described organic solvent is the mixture of any one or at least two kinds in ethyl acetate, dichloromethane or chloroform.
In the preparation method of support type hemoglobin compositions of the present invention, the aqueous solution of step (2) described surfactant is the mixture of polyvinyl alcohol water solution or polyvinyl alcohol water solution and propylene glycol block polyether aqueous solution, and wherein said propylene glycol block polyether can be any one in L31, L35, F38, L42, L43, L44, L45, L61, L62, L63, L64, P65 or F68.
Preferably, the aqueous solution of step of the present invention (2) described surfactant is the mixture of polyvinyl alcohol water solution and propylene glycol block polyether aqueous solution, in described mixture, the volume ratio of polyvinyl alcohol water solution and propylene glycol block polyether aqueous solution is 1:1-9:1, such as 1:1,2:1,3:1,3:2,4:1,4:2,4:3,5:1,5:2,5:3,5:4,6:1,6:5,7:1,7:2,7:3,7:4,7:5,7:6,8:1,8:3,8:5,8:7 or 9:1, be preferably 7:3; Preferably, the concentration of described polyvinyl alcohol water solution and propylene glycol block polyether aqueous solution is 2wt%.
In the preparation method of support type hemoglobin compositions of the present invention, the mass ratio of step (1) described carrier PEG-PLGA block copolymer and hemoglobin is 5:4-5:12, such as 5:4,1:1,5:6,5:7,5:8,5:9,1:2,5:11 or 5:12, be preferably 5:6.
In the preparation method of support type hemoglobin compositions of the present invention, the concentration of step (1) described haemoglobin aqueous solution is 60-240mg/mL, such as 60mg/mL, 70mg/mL, 80mg/mL, 90mg/mL, 100mg/mL, 110mg/mL, 120mg/mL, 140mg/mL, 160mg/mL, 180mg/mL, 190mg/mL, 200mg/mL, 210mg/mL, 220mg/mL, 230mg/mL or 240mg/mL, be preferably 120mg/mL; Preferably, step (1) described PEG-PLGA block copolymerization substrate concentration is 10-30mg/mL, such as 10mg/mL, 11mg/mL, 12mg/mL, 15mg/mL, 17mg/mL, 19mg/mL, 20mg/mL, 22mg/mL, 24mg/mL, 26mg/mL, 27mg/mL, 28mg/mL, 29mg/mL or 30mg/mL, be preferably 20mg/mL; Preferably, relative to PEG-PLGA di-block copolymer solution described in 2mL, the amount of aqueous solution used of described surfactant is 10mL.
In the preparation method of support type hemoglobin compositions of the present invention, the described ultrasonic ultrasonic cell disintegration instrument that utilizes of step (1) carries out under 80w in ice-water bath, and ultrasonic time is 3-5min; Preferably, the described ultrasonic ultrasonic cell disintegration instrument that utilizes of step (2) carries out under 250w in ice-water bath, and ultrasonic time is 3-5min; Preferably, step (3) described centrifugal rotational speed is 6,000-13,000rpm, and centrifugation time is 8-15min.
As optimal technical scheme, the preparation method of support type hemoglobin compositions of the present invention, specifically comprises the following steps:
(1) PEG-PLGA block copolymer is dissolved in ethyl acetate, its concentration is made to be 20mg/mL, be the aqueous solution of the hemoglobin of 120mg/mL again by 2mL PEG-PLGA block copolymer solution and 0.4mL concentration, utilize ultrasonic cell disintegration instrument under 80w in ice-water bath ultrasonic 3min, formed colostrum;
(2) just add the aqueous solution of 10mL surfactant in Ruzhong to step (1) gained, mixing, to utilize under ultrasonic cell disintegration instrument 250w ultrasonic 5min in ice-water bath, form emulsion;
(3) remove ethyl acetate, with the centrifugal 10min of 13,000rpm rotating speed, obtain support type hemoglobin compositions;
The aqueous solution of wherein said surfactant is the mixture of polyvinyl alcohol water solution and propylene glycol block polyether aqueous solution, the volume ratio of described polyvinyl alcohol water solution and propylene glycol block polyether aqueous solution is 7:3, and the concentration of described polyvinyl alcohol water solution and propylene glycol block polyether aqueous solution is 2wt%.
On the other hand, the invention provides the support type hemoglobin compositions prepared by preparation method of the present invention.Described in the support type hemoglobin compositions that the present invention prepares, carrier PEG-PLGA block copolymer is 43.75-75.00% to the envelop rate of hemoglobin, such as 43.75%, 44.60%, 50.89%, 53.00%, 54.00%, 57.92%, 62.00%, 64.17%, 70.00%, 72.00% or 75.00%, be preferably 55%.
The mean diameter of the nanoparticle that the support type hemoglobin compositions that the present invention prepares is formed is 143.1-267.4nm, such as 143.1nm, 150.6nm, 155.9nm, 159.1nm, 168.4nm, 170.2nm, 175.2nm, 180.5nm, 185.3nm, 190.6nm, 196.3nm, 200.6nm, 205.8nm, 216.8nm, 230.9nm, 251.8nm or 267.4nm.
Support type hemoglobin compositions prepared by the present invention can be used as the carrier of oxygen of hemoglobin, as blood substitute, because it has less particle diameter, the longer time can be stablized in blood plasma, avoid hemoglobin in transfusion procedure directly with surrounding tissue and other composition direct effects of blood and bring vasoconstriction, nephrotoxicity problem, also can avoid hemoglobin oxidized caused by serious untoward reaction.
The present invention has following beneficial effect:
(1) the present invention adopts two newborn legal system for support type hemoglobin compositions, method therefor is simple, and selected carrier material not only abundant raw material, and there is low cost, biodegradable, good biocompatibility and bioaffinity, nontoxic, be easy to the advantages such as chemical modification;
(2) support type hemoglobin compositions self assembly in aqueous medium that prepared by the present invention forms nanoparticle, there is the mean diameter of 143.1-267.4nm, particle size distribution is narrower, and there is nucleocapsid structure, in monodispersed regular spheroid, soilless sticking phenomenon, system is more stable, nanoparticle macrophage phagocytic can be prevented, the circulation time in extension body;
(3) the present invention passes through by hemoglobin load in the carrier, both solved hemoglobin and surrounding tissue and other composition direct effects of blood and bring vasoconstriction, nephrotoxicity problem, also can avoid the oxidized caused serious untoward reaction of hemoglobin;
(4) the support type hemoglobin compositions that prepared by the present invention possesses to take puts oxygen function, the not chromotoxic secondary structure of loading process, approximate rheological behavior is had with whole blood, stability is high, the seepage that hemoglobin is a large amount of can not be caused, and possess good blood compatibility, do not cause the change of whole blood coagulation function, erythrocyte aggregation can not be caused, do not cause haemolysis, can be used as blood substitute.
Accompanying drawing explanation
Fig. 1 is the transmission electron microscope picture of the PEG-PLGA block copolymer nano particle of load hemoglobin prepared by the embodiment of the present invention 1;
Fig. 2 is the grain size distribution of the PEG-PLGA block copolymer nano particle of load hemoglobin prepared by the embodiment of the present invention 1;
Fig. 3 is the change of size in the PEG-PLGA block copolymer nano system put procedure of load hemoglobin prepared by the embodiment of the present invention 1;
Fig. 4 is the visible ray spectrogram of the PEG-PLGA block copolymer nano particle of load hemoglobin prepared by the embodiment of the present invention 3;
Fig. 5 is that the taking of PEG-PLGA block copolymer nano particle of load hemoglobin prepared by the embodiment of the present invention 3 puts oxygen curve;
Fig. 6 is the secondary structure of the PEG-PLGA block copolymer nano particle of load hemoglobin prepared by the embodiment of the present invention 3;
Fig. 7 is the rheological behavior of the PEG-PLGA block copolymer nano particle of load hemoglobin prepared by the embodiment of the present invention 3;
Fig. 8 is that the PEG-PLGA block copolymer nano particle of load hemoglobin prepared by the embodiment of the present invention 3 is on the impact of whole blood rheological characteristic;
Fig. 9 is that the PEG-PLGA block copolymer nano particle of load hemoglobin prepared by the embodiment of the present invention 3 is on the impact of erythrocyte aggregation.
Detailed description of the invention
Technical scheme of the present invention is further illustrated below by detailed description of the invention.Those skilled in the art should understand, described embodiment is only help to understand the present invention, should not be considered as concrete restriction of the present invention.
The present invention passes through two newborn legal system for support type hemoglobin compositions nanoparticle, due to the PEG-PLGA block copolymer nanoparticle that self assembly is formed in aqueous medium, assemble the kernel forming particle due to hydrophobic interaction between hydrophobic segment wherein, hydrophilic segment then forms the shell of particle, therefore can load cancer therapy drug, protein, RNA isoreactivity composition.
key instrument and reagent
Ultrasonic cell disintegration instrument: NingBo XinZhi Biology Science Co., Ltd, model: Scientz-II D
Rotary Evaporators: Shanghai Yarong Biochemical Instrument Plant, model: RE52CS-2
Transmission electron microscope: NEC company, model is JEM-200CX
Laser particle analyzer: Malvern Instr Ltd., Zetasizer Nano ZS, Britain
Ultraviolet spectrophotometer: power & light company of the U.S., He λ ios β
Blood oxygen analysis instrument: Beijing Kai Zheng biological engineering Co., Ltd, KZ-176
Circular dichroism spectra: Jia Sike (Shanghai) trade Co., Ltd, J-810
Semi-automatic hemorheology tester: Bo Laite (China), BT-300
Inverted microscope, COIC, XDS-1B
PEG-PLGA block copolymer material is purchased from Jinan Dai Gang biological engineering company limited, wherein the weight average molecular weight of Polyethylene Glycol segment is 5000Da, the weight average molecular weight of polylactic-co-glycolic acid segment is 45000Da, and polylactic-co-glycolic acid segment is PLGA75/25.Polyvinyl alcohol (PVA) is purchased from chemical reagent GuoYao Stock Co., Ltd (Sinopharm Chemical Reagent Co., Ltd), propylene glycol block polyether F68 and F38 is purchased from this reagent company limited of Adama (Adamas Reagent Co., Ltd).
Hemoglobin: extract blood from the bull, ox (laboratory animal is from Beijing beautiful the earth agricultural Tourism Garden of Ecological) of health, make erythrocyte membrane osmotic bursting by centrifugal, obtain hemoglobin solutions, use Normal Saline dilution, obtain the haemoglobin aqueous solution of variable concentrations, for subsequent use.In hemoglobin prepared by the method, ferrihemoglobin content is less than 3%.
embodiment 1
Prepare the PEG-PLGA block copolymer nano particle of negative loaded with hemoglobin in the present embodiment by the following method, the method comprises the following steps:
(1) haemoglobin aqueous solution be ethyl acetate solution and the 0.4mL concentration of PEG-PLGA block copolymer of 10mg/mL by 2mL concentration being 120mg/mL mixes, concussion makes their mix homogeneously, in ice-water bath, by above-mentioned solution ultrasonic cell disintegration instrument ultrasonic 3min under 80w, form colostrum;
(2) F683mL of PVA 7mL and 2wt% of 2wt% is then got, slowly be added drop-wise in step (1) described colostric fluid with dropper, concussion makes its mix homogeneously, in ice-water bath, by above-mentioned solution ultrasonic cell disintegration instrument ultrasonic 5min under 250w, form emulsion;
(3) with Rotary Evaporators removing ethyl acetate organic solvent, with the centrifugal 10min of 13,000rpm rotating speed, removing supernatant, obtains the PEG-PLGA block copolymer nano particle of negative loaded with hemoglobin.
The character of the PEG-PLGA block copolymer nano particle of the negative loaded with hemoglobin prepared is detected:
The hemoglobin concentration not wrapping into nanoparticle is measured by cyanmethemoglobin method, according to formula EE%=(tHb-fHb)/tHb × 100%, investigate the envelop rate of the PEG-PLGA block copolymer nano grain carrying hemoglobin, wherein tHb is total hemoglobin gross mass, fHb is the hemoglobin gross mass not wrapping into nanoparticle, and envelop rate is 43.75% as calculated.
The PEG-PLGA block copolymer nano particle utilizing transmission electron microscope observation to obtain, tentatively determines the distribution situation of nanometer particle size, as shown in Figure 1.Fig. 1 shows that the PEG-PLGA block copolymer nano particle of the negative loaded with hemoglobin prepared is spheroidal particle, and these particle dispersion are even, and soilless sticking phenomenon.
Utilize laser particle analyzer to detect the PEG-PLGA block copolymer nano particle obtained, measure the particle size distribution of nanoparticle, as shown in Figure 2.Fig. 2 shows that the particle size distribution of the PEG-PLGA block copolymer nano particle of the negative loaded with hemoglobin prepared is narrower, and good dispersion, mean diameter is 143.1nm.
The PEG-PLGA block copolymer nano particle of the negative loaded with hemoglobin prepared by the present embodiment places 7 days, laser particle analyzer is utilized to monitor its nanometer particle size every day, to determine the stable case of the PEG-PLGA block copolymer nano particle of prepared negative loaded with hemoglobin, result as shown in Figure 3, can draw from result, the PEG-PLGA block copolymer nano particle of the negative loaded with hemoglobin of preparation has higher stability.
embodiment 2
Prepare the PEG-PLGA block copolymer nano particle of negative loaded with hemoglobin in the present embodiment by the following method, the method comprises the following steps:
(1) haemoglobin aqueous solution be dichloromethane solution and the 0.4mL concentration of PEG-PLGA block copolymer of 15mg/mL by 2mL concentration being 120mg/mL mixes, concussion makes their mix homogeneously, in ice-water bath, by above-mentioned solution ultrasonic cell disintegration instrument ultrasonic 3min under 100w, form colostrum;
(2) then get the PVA 10mL of 2wt%, be slowly added drop-wise in above-mentioned colostric fluid with dropper, concussion makes its mix homogeneously, in ice-water bath, by above-mentioned solution ultrasonic cell disintegration instrument ultrasonic 5min under 200w, forms emulsion;
(3) with Rotary Evaporators removing dichloromethane organic solvent, with the centrifugal 12min of 10,000rpm rotating speed, removing supernatant, obtains the PEG-PLGA block copolymer nano particle of negative loaded with hemoglobin.
Known by the test identical with embodiment 1, the PEG-PLGA block copolymer nano particle of negative loaded with hemoglobin prepared by the present embodiment has regular spheroidal structure, the mean diameter of the nanoparticle obtained is 168.4nm, and envelop rate is 50.89% as calculated.
embodiment 3
Prepare the PEG-PLGA block copolymer nano particle of negative loaded with hemoglobin in the present embodiment by the following method, the method comprises the following steps:
(1) haemoglobin aqueous solution be PEG-PLGA block copolymer ethyl acetate solution and the 0.4mL concentration of 20mg/mL by 2mL concentration being 120mg/mL mixes, concussion makes their mix homogeneously, in ice-water bath, by above-mentioned solution ultrasonic cell disintegration instrument ultrasonic 3min under 80w, form colostrum;
(2) get the F683mL of PVA 7mL and 2wt% of 2wt%, be slowly added drop-wise in above-mentioned colostric fluid with dropper, concussion makes its mix homogeneously, in ice-water bath, above-mentioned solution is used again ultrasonic cell disintegration instrument ultrasonic 5min under 250w, forms emulsion;
(3) with Rotary Evaporators removing ethyl acetate organic solvent, with the centrifugal 10min of 13,000rpm rotating speed, removing supernatant, obtains the PEG-PLGA block copolymer nano particle of negative loaded with hemoglobin.
Known by the test identical with embodiment 1, PEG-PLGA block copolymer nano particle prepared by the present embodiment has regular spheroidal structure, and the mean diameter of the nanoparticle obtained is 196.3nm, and envelop rate is 55% as calculated.
Hemoglobin characteristic spectrum situation of change in nanoparticle is observed by the full wavelength scanner of the PEG-PLGA block copolymerization matter sample of negative loaded with hemoglobin prepared the present embodiment, judge by the characteristic peak misalignment observing 400-430nm place the situation that hemoglobin is oxidized, result as shown in Figure 4.
Oxygen curve and P50 (partial pressure of oxygen number when referring to hemoglobin 50% oxygen saturation is put by measuring taking of the PEG-PLGA block copolymerization matter sample of negative loaded with hemoglobin prepared by the present embodiment, reflection hemoglobin is taken and is put oxygen function, value is lower shows that its oxygen affinity is higher) determine the oxygen affinity of this sample, thus determine that the taking of PEG-PLGA block copolymer nano particle of carrying hemoglobin puts oxygen function, result is as shown in Figure 5.
The PEG-PLGA block copolymerization matter sample of negative loaded with hemoglobin is added in circular dichroism spectrometer test trough, measure the circular dichroism spectrogram at 190-240nm place, thus the preparation process obtaining this sample is on the impact of hemoglobin secondary structure, result as shown in Figure 6.
The PEG-PLGA block copolymerization matter sample 200 μ L being born loaded with hemoglobin joins in evolute hemorheology tester, and detecting its shear rate is 1 ~ 210S
-1rheological behavior, result is as shown in Figure 7.
The PEG-PLGA block copolymerization matter sample 200 μ L being born loaded with hemoglobin mixes with the whole blood of 800 μ L, joins evolute hemorheology tester, and detecting its shear rate is 1 ~ 210S
-1rheological behavior, result is as shown in Figure 8.
Mixed with red cell suspension by the PEG-PLGA block copolymerization matter sample of negative loaded with hemoglobin, by this nanoparticle of observation by light microscope on the impact of erythrocyte aggregation, result as shown in Figure 9.
Fig. 4 to Fig. 9 shows: the PEG-PLGA di-block copolymer nanoparticle sample of the load hemoglobin of embodiments of the invention possesses to take puts oxygen function, its P50 is 14.59, the not chromotoxic secondary structure of loading process, approximate rheological behavior is had with whole blood, stability is high, can not cause the seepage that hemoglobin is a large amount of.Possess good blood compatibility, can not erythrocyte aggregation be caused, can be used as blood substitute.
embodiment 4
Prepare the PEG-PLGA block copolymer nano particle of negative loaded with hemoglobin in the present embodiment by the following method, the method comprises the following steps:
(1) haemoglobin aqueous solution be PEG-PLGA di-block copolymer ethyl acetate solution and the 0.4mL concentration of 30mg/mL by 2mL concentration being 120mg/mL mixes, concussion makes their mix homogeneously, in ice-water bath, by above-mentioned solution ultrasonic cell disintegration instrument ultrasonic 5min under 80w, form colostrum;
(2) get the F681mL of PVA 9mL and 2wt% of 2wt%, be slowly added drop-wise in above-mentioned colostric fluid with dropper, concussion makes its mix homogeneously, in ice-water bath, by above-mentioned solution ultrasonic cell disintegration instrument ultrasonic 3min under 250w, forms emulsion;
(3) with Rotary Evaporators removing ethyl acetate organic solvent, with the centrifugal 15min of 6,000rpm rotating speed, removing supernatant, obtains the PEG-PLGA di-block copolymer nanoparticle of loaded with hemoglobin.
Known by the test identical with embodiment 1, the PEG-PLGA block copolymer nano particle of negative loaded with hemoglobin prepared by the present embodiment has regular spheroidal structure, the mean diameter of the nanoparticle obtained is 248.1nm, and envelop rate is 64.17% as calculated.
embodiment 5
Prepare the PEG-PLGA block copolymer nano particle of negative loaded with hemoglobin in the present embodiment by the following method, the method comprises the following steps:
(1) PEG-PLGA block copolymer ethyl acetate and chloroform are dissolved, concentration is 15mg/mL, get the haemoglobin aqueous solution that this solution 2mL and 0.4mL concentration are 60mg/mL to mix, concussion makes their mix homogeneously, in ice-water bath, by above-mentioned solution ultrasonic cell disintegration instrument ultrasonic 5min under 80w, form colostrum;
(2) get the F382mL of PVA 8mL and 2wt% of 2wt%, be slowly added drop-wise in above-mentioned colostric fluid with dropper, concussion makes its mix homogeneously, in ice-water bath, by above-mentioned solution ultrasonic cell disintegration instrument ultrasonic 5min under 200w, forms emulsion;
(3) with Rotary Evaporators removing ethyl acetate and chloroform organic solvent; With the centrifugal 8min of 13,000rpm rotating speed, removing supernatant, obtains the PEG-PLGA di-block copolymer nanoparticle of loaded with hemoglobin.
Known by the test identical with embodiment 1, the PEG-PLGA block copolymer nano particle of negative loaded with hemoglobin prepared by the present embodiment has regular spheroidal structure, the mean diameter that the nanoparticle obtained is large is 159.1nm, and envelop rate is 75.00% as calculated.
embodiment 6
Prepare the PEG-PLGA block copolymer nano particle of negative loaded with hemoglobin in the present embodiment by the following method, the method comprises the following steps:
(1) be that the PEG-PLGA di-block copolymer three of 20mg/mL is filtered the haemoglobin aqueous solution that dichloromethane and 0.4mL concentration are 240mg/mL and mixed by 2mL concentration, concussion makes their mix homogeneously, in ice-water bath, by above-mentioned solution ultrasonic cell disintegration instrument ultrasonic 3min under 90w, form colostrum;
(2) then get the F385mL of PVA 5mL and 2wt% of 2wt%, be slowly added drop-wise in above-mentioned colostric fluid with dropper, concussion makes its mix homogeneously, in ice-water bath, by above-mentioned solution ultrasonic cell disintegration instrument ultrasonic 5min under 230w, forms emulsion;
(3) with Rotary Evaporators removing ethyl acetate organic solvent, with the centrifugal 10min of 13,000rpm rotating speed, removing supernatant, obtains the PEG-PLGA di-block copolymer nanoparticle of loaded with hemoglobin.
Known by the test identical with embodiment 1, the PEG-PLGA di-block copolymer nanoparticle of negative loaded with hemoglobin prepared by the present embodiment has regular spheroidal structure; The mean diameter that the nanoparticle obtained is large is 267.4nm, and envelop rate is 44.60% as calculated.
Therefore, the support type hemoglobin compositions prepared by the present invention, narrow diameter distribution, and there is nucleocapsid structure, and in monodispersed regular spheroid, soilless sticking phenomenon, system is more stable, can prevent nanoparticle macrophage phagocytic, the circulation time in extension body; And possess to take and put oxygen function, the not chromotoxic secondary structure of loading process, have approximate rheological behavior with whole blood, stability is high, can not cause the seepage that hemoglobin is a large amount of.And possess good blood compatibility, do not cause the change of whole blood coagulation function, erythrocyte aggregation can not be caused, do not cause haemolysis, can be used as blood substitute.
Applicant states, the present invention illustrates support type hemoglobin compositions of the present invention and preparation method thereof by above-described embodiment, but the present invention is not limited to above-mentioned specific embodiment.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 raw material selected by the present invention, all drops within protection scope of the present invention and open scope.
Claims (10)
1. a preparation method for support type hemoglobin compositions, is characterized in that, said method comprising the steps of:
(1) carrier PEG-PLGA block copolymer is dissolved in organic solvent, then by the aqueous solution of itself and hemoglobin, ultrasonic, forms colostrum;
(2) just the aqueous solution of surfactant is added in Ruzhong to step (1) gained, mixing, ultrasonic, form emulsion;
(3) organic solvent is removed, centrifugal, obtain support type hemoglobin compositions.
2. preparation method according to claim 1, is characterized in that, the structural formula of step (1) described carrier PEG-PLGA block copolymer is as follows:
Preferably, in described PEG-PLGA block copolymer, the weight average molecular weight of Polyethylene Glycol segment is 1000-5000Da, and the weight average molecular weight of polylactic-co-glycolic acid segment is 4500-45000Da;
Preferably, described polylactic-co-glycolic acid segment is formed by lactic acid and hydroxyacetic acid two kinds of monomer polymerizations, and the mol ratio of wherein said lactic acid and hydroxyacetic acid two kinds of monomers is 75:25 or 50:50.
3. preparation method according to claim 1 and 2, is characterized in that, step (1) described organic solvent is the mixture of any one or at least two kinds in ethyl acetate, dichloromethane or chloroform.
4. the preparation method according to any one of claim 1-3, is characterized in that, the aqueous solution of step (2) described surfactant is the mixture of polyvinyl alcohol water solution or polyvinyl alcohol water solution and propylene glycol block polyether aqueous solution.
5. the preparation method according to any one of claim 1-4, it is characterized in that, the aqueous solution of step (2) described surfactant is the mixture of polyvinyl alcohol water solution and propylene glycol block polyether aqueous solution, in described mixture, the volume ratio of polyvinyl alcohol water solution and propylene glycol block polyether aqueous solution is 1:1-9:1, is preferably 7:3;
Preferably, the concentration of described polyvinyl alcohol water solution and propylene glycol block polyether aqueous solution is 2wt%.
6. the preparation method according to any one of claim 1-5, is characterized in that, the mass ratio of step (1) described carrier PEG-PLGA block copolymer and hemoglobin is 5:4-5:12, is preferably 5:6.
7. the preparation method according to any one of claim 1-6, is characterized in that, the concentration of aqueous solution of step (1) described hemoglobin is 60-240mg/mL, is preferably 120mg/mL;
Preferably, step (1) described PEG-PLGA block copolymerization substrate concentration is 10-30mg/mL, is preferably 20mg/mL;
Preferably, relative to PEG-PLGA block copolymer solution described in 2mL, the amount of aqueous solution used of described surfactant is 10mL.
8. the preparation method according to any one of claim 1-7, is characterized in that, the described ultrasonic ultrasonic cell disintegration instrument that utilizes of step (1) carries out under 80-100w in ice-water bath, and ultrasonic time is 3-5min;
Preferably, the described ultrasonic ultrasonic cell disintegration instrument that utilizes of step (2) carries out under 200-250w in ice-water bath, and ultrasonic time is 3-5min;
Preferably, step (3) described centrifugal rotational speed is 6,000-13,000rpm, and centrifugation time is 8-15min.
9. the preparation method according to any one of claim 1-8, is characterized in that, said method comprising the steps of:
(1) PEG-PLGA block copolymer is dissolved in ethyl acetate, its concentration is made to be 20mg/mL, by the aqueous solution that 2mL PEG-PLGA block copolymer solution and 0.4mL concentration are the hemoglobin of 120mg/mL, utilize ultrasonic cell disintegration instrument under 80w in ice-water bath ultrasonic 3min, formed colostrum;
(2) just add the aqueous solution of 10mL surfactant in Ruzhong to step (1) gained, mixing, to utilize under ultrasonic cell disintegration instrument 250w ultrasonic 5min in ice-water bath, form emulsion;
(3) remove ethyl acetate, with the centrifugal 10min of 13,000rpm rotating speed, obtain support type hemoglobin compositions;
The aqueous solution of wherein said surfactant is the mixture of polyvinyl alcohol water solution and propylene glycol block polyether aqueous solution, the volume ratio of described polyvinyl alcohol water solution and propylene glycol block polyether aqueous solution is 7:3, and the concentration of described polyvinyl alcohol water solution and propylene glycol block polyether aqueous solution is 2wt%.
10. the support type hemoglobin compositions that the preparation method according to any one of claim 1-9 prepares.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510069797.5A CN104644614B (en) | 2015-02-10 | 2015-02-10 | A kind of support type hemoglobin compositions and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510069797.5A CN104644614B (en) | 2015-02-10 | 2015-02-10 | A kind of support type hemoglobin compositions and preparation method thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104644614A true CN104644614A (en) | 2015-05-27 |
| CN104644614B CN104644614B (en) | 2017-11-24 |
Family
ID=53236589
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510069797.5A Expired - Fee Related CN104644614B (en) | 2015-02-10 | 2015-02-10 | A kind of support type hemoglobin compositions and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104644614B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108893488A (en) * | 2018-06-19 | 2018-11-27 | 刘志强 | A kind of recombinant plasmid, DNA vaccination and its preparation method and application |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20050159339A1 (en) * | 2002-03-25 | 2005-07-21 | Acharya Seetharama A. | Pegylated non-hypertensive hemoglobins, methods of preparing same, and uses thereof |
| CN101732723A (en) * | 2009-12-30 | 2010-06-16 | 上海市肿瘤研究所 | Polyethylene glycol-poly(lactic-co-glycolic acid)-polylysine nano-delivery system, preparation method and application thereof |
-
2015
- 2015-02-10 CN CN201510069797.5A patent/CN104644614B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20050159339A1 (en) * | 2002-03-25 | 2005-07-21 | Acharya Seetharama A. | Pegylated non-hypertensive hemoglobins, methods of preparing same, and uses thereof |
| CN101732723A (en) * | 2009-12-30 | 2010-06-16 | 上海市肿瘤研究所 | Polyethylene glycol-poly(lactic-co-glycolic acid)-polylysine nano-delivery system, preparation method and application thereof |
Non-Patent Citations (3)
| Title |
|---|
| JINGYAN LI ET AL: "PLA/PLGA nanoparticles for delivery of drugsacross the blood-brain barrier", 《NANOTECHNOL REVIEW》 * |
| 刘英华: "两亲性纳米载药***的制备与性能研究", 《中国优秀硕士学位论文全文数据库 工程科技 I 辑》 * |
| 高焕民: "《纳米医学》", 31 May 2006 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108893488A (en) * | 2018-06-19 | 2018-11-27 | 刘志强 | A kind of recombinant plasmid, DNA vaccination and its preparation method and application |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104644614B (en) | 2017-11-24 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Hao et al. | Targeted imaging and chemo‐phototherapy of brain cancer by a multifunctional drug delivery system | |
| Ren et al. | Preparation and therapeutic efficacy of polysorbate-80-coated amphotericin B/PLA-b-PEG nanoparticles | |
| Su et al. | Microfluidic synthesis of manganese-alginate nanogels with self-supplying H2O2 capability for synergistic chemo/chemodynamic therapy and boosting anticancer immunity | |
| CN110302175B (en) | Lipoic acid nanocapsule containing hydrophobic cavity and preparation method and application thereof | |
| Dai et al. | Cellulose-graft-poly (l-lactic acid) nanoparticles for efficient delivery of anti-cancer drugs | |
| Zhang et al. | PEGylated nanostructured lipid carriers loaded with 10‐hydroxycamptothecin: an efficient carrier with enhanced anti‐tumour effects against lung cancer | |
| CN105497912A (en) | Preparation method and application of erythrocyte membrane-coated acid-sensitive polymer prodrug nano drug delivery system | |
| Xuan et al. | Bismuth particles imbedded degradable nanohydrogel prepared by one-step method for tumor dual-mode imaging and chemo-photothermal combined therapy | |
| Dai et al. | Self-assembled serum albumin–poly (l-lactic acid) nanoparticles: a novel nanoparticle platform for drug delivery in cancer | |
| CN108619094A (en) | A kind of nanometer formulation and preparation method thereof of anticancer natural product gambogicacid | |
| Xiao et al. | A facile strategy for fine-tuning the stability and drug release of stimuli-responsive cross-linked micellar nanoparticles towards precision drug delivery | |
| Chen et al. | Promotion of the anticancer activity of curcumin based on a metal–polyphenol networks delivery system | |
| CN112999153A (en) | Nano micelle carrying chemotherapeutic drug/photosensitizer and preparation method and application thereof | |
| Zhang et al. | Bovine serum albumin-based and dual-responsive targeted hollow mesoporous silica nanoparticles for breast cancer therapy | |
| CN102139113B (en) | Novel pharmaceutical solubilization carrier and preparation method and application thereof | |
| CN107049955A (en) | A kind of multistage targeting hyaluronan nanoparticle for carrying methotrexate (MTX) and preparation method thereof | |
| Kim et al. | Plant-inspired Pluronic–gallol micelles with low critical micelle concentration, high colloidal stability, and protein affinity | |
| Cheng et al. | A sequentially responsive cascade nanoplatform for increasing chemo-chemodynamic therapy | |
| Zhou et al. | Biocompatibility and characteristics of chitosan/cellulose acetate microspheres for drug delivery | |
| CN104644614A (en) | Supported hemoglobin composition and preparation method thereof | |
| Luu et al. | Nanoencapsulation of Chromolaena odorata extract using pluronic F127 as an effectively herbal delivery system for wound healing | |
| CN104784117A (en) | Curcumin mixed micelle oral preparation and preparation method thereof | |
| CN104644613B (en) | PEG-PLGA wraps up hemoglobin | |
| WO2023193389A1 (en) | Resveratrol-lecithin nanoparticle, method for preparing same, and use thereof | |
| CN103006546A (en) | Preparation and application of carbon nano tube-containing thermo-sensitive type gel entrapping for indissolvable drug |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20171124 |