CN105343890B - A kind of heparin or the graphene oxide of its salt modification and preparation method and application - Google Patents
A kind of heparin or the graphene oxide of its salt modification and preparation method and application Download PDFInfo
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Abstract
The present invention discloses a kind of heparin or the graphene oxide of its salt modification and preparation method and application, and the graphene oxide after modification can be as the carrier material of medicine particularly cancer therapy drug.Using pH sensitivity principles using adipic dihydrazide as connection molecule, or using redox principle using cystamine as connection molecule, graphene oxide is chemically modified with heparin, the advantages of synergy antitumor with the increase water solubility of graphene oxide, cancer therapy drug, solves the problems such as graphene oxide water solubility is poor, easily assembles.
Description
Technical field
The present invention relates to pharmaceutical synthesis field, and in particular to a kind of graphene oxide modified for heparin or its salt and its
Preparation method and application.
Background technology
Graphene oxide (Graphene oxide, GO, see Fig. 1) is the derivative of the graphene with monoatomic layer structure
(Carbon,2011,49:1126-1132).Compared with the graphene (Graphene) on surface only containing carbon-carbon double bond, oxidation
Containing oxy radicals such as substantial amounts of hydroxyl, epoxy radicals, carboxyls in graphene self structure, and there is preferable hydrophily, can be wide
It is general be applied to it is biomedical in (J Control Release, 2014,173:75-88).Because graphene oxide is monoatomic layer
Structure, there is larger specific surface area (to have document to be measured as 736.6m2/ g, is shown in Langmuir, 2013,29:13443-
13448), its two sides can all be combined by covalent, noncovalent interaction with medicine, therefore have higher drug load amount.It can
By stronger physisorption and aromatic rings class medicine Non-covalent binding, some insoluble drugs can be delivered, particularly pair
Transport and have great importance inside most of slightly solubility cancer therapy drug (Science, 2004,306:666).Graphene oxide
For hydroaropic substance, there is preferable biocompatibility.Research is found, is a kind of quite safe in cellular level graphene oxide
Material, without obvious CDCC (Toxicol Lett, 2011,200:201), therefore graphene oxide is as medicine
The carrier of targeting conveying is increasingly paid attention to by scientific research personnel, and presently disclosed patent is more.
As patent CN201210509075 reports a kind of graphene oxide of glucan chemical graft ferroheme modification, use
In diagnosis and treatment that drug delivery etc. is biomedical.Patent CN201310450626 reports a kind of graphene oxide-boron and changed
Property phenolic resin preparation method, not only maintain the structure and performance of graphene, and the part oxygen-containing functional group energy retained
Solve the problems such as graphene dispersion, dissolubility and poor in processability well.Patent CN201310185529 reports a kind of oxygen
The preparation method of graphite alkene and aqueous polyurethane nano composite, the hydrophily of graphene oxide layer can be reduced, improved
Its dispersiveness in organic solvent and the intermiscibility between polymer.Patent CN201210038641 reports a kind of oxidation
Graphene-sanguinarine compound and preparation method thereof.In the compound, by non-common between graphene oxide carrier and sanguinarine
Valency π-π effects combine, and can greatly improve the solubility and stability of sanguinarine, can also make it have targeting, and be expected to conduct
A kind of anticancer preparation having a high potential is widely used.Patent CN201110361574 reports a kind of polystyrene graft oxidation
The method of graphene.The method that patent CN201110227223 reports polyethyleneglycol modified graphene oxide.
Graphene oxide can turn after reducing agent (such as hydrazine, sodium borohydride, 1,2,3,-thrihydroxy-benzene, ascorbic acid, thiocarbamide) reduction
It is changed into reproducibility graphene oxide (reduced GO, rGO, see Fig. 2).Compared with graphene oxide, rGO eliminates more contain
Oxygen groups, planar structure is maintained, but there is the characteristics of irreversible aggrengation, or lacking macromolecule dispersing agent or surface work
Be piled under conditions of property agent graphite (Carbon, 2012,50:3210-3228).Not only drug carrying ability is significantly by rGO
Improve, and can absorb the ability of the infrared ray in the range of 650-900nm and produce heat (photo-thermal effect) and greatly enhance (J Am
Chem Soc,2009,131:11027-11032).Such as Doxorubicin, its drugloading rate improve 10 times (ACS Nano,
2013,7:6735-6746), it is 0.6W/cm in 808nm near infrared light and energy2Illumination under, its photo-thermal effect adds 6
Times (J Am Chem Soc, 2011,133:6825-6831).The higher load property of medicine and stronger photo-thermal effect using rGO, greatly
The earth enhances antitumor targeting ability and therapeutic effect, there is also more report at present.
Document report, to increase the transport efficacy in target cancer cells of cancer therapy drug, reduce the poison to other normal cells
Property, its curative effect is improved, graphene oxide is needed before pharmaceutical carrier by certain chemical modification, its active anticancer of competence exertion
(such as Nano Lett.2010,10:3318-3323, Adv Mater.2012,24:1722-1728, ACS Nano.2011,5:
7000-7009, Biomaterials.2014,35:4986-4995 etc.).Such as graphene oxide after polyethyleneglycol modified its water
Dissolubility further enhances (J Am Chem Soc.2008；130:10876-10877).And for example under certain condition, stone will be aoxidized
Black alkene and sodium chloroacetate (ClCH2COONa) mix, the hydroxyl in graphene oxide, epoxy radicals, ester group is converted to carboxyl, then
It is sulfonated, graphene oxide derivative of the generation containing a large amount of sulfonic groups and carboxyl, so as to improve graphene oxide in physiology
Stability (Small, 2010,6 (4) in solution:37).Carboxyl (COOH) in the derivative can be with the amino in folic acid
(NH2) covalent bond generation folic acid-graphene oxide conjugate.Because folic acid alternative is identified by folacin receptor, and folic acid
Acceptor great expression in human breast cancer cell, therefore, the medicine being supported on the graphene oxide of folic acid surface modification can be real
Existing target administration (Nano Res, 2008,1:203-212).By the pi-pi accumulation and hydrophobic effect of graphene oxide, this is conjugated
Thing can be by cancer therapy drug camptothecine (DNA topoisomerase I inhibitors), Doxorubicin (DNA Topoisomerase II inhibitors) thing
Reason absorption forms the compound containing cancer therapy drug in surface of graphene oxide.The compound through blood (pH 7.35~
7.45) position (such as human breast cancer cell) of folacin receptor great expression is transported, will through receptor-mediated endocytosis
Compound containing anticarcinogen is transported in cancer cell, in endocytosis body (pH 5.0~6.5) and the acid of lysosome (pH 4.5~5.0)
Property destruction, discharge cancer therapy drug (Biomaterials, 2013,34:3647-3657).Further, since tumour cell is given birth to
It is long fast, the acidic metabolites such as lactic acid more more than normal structure, therefore, tumour cell periphery are generated in breeding
Tissue fluid pH value is lower than normal cell, shows faintly acid (pH6.5~7.2, Biomacromolecules 2009；10(7):1727-
1735；J Am Chem Soc 2005,127(51):17982-17983).And anticarcinogen is mostly alkaline drug, such as camptothecine
PKa=8.63 (Curr Med Chem.2011,18 (9):1367-1372), the pKa=8.22 (Analytical of Doxorubicin
Profiles of Drug Substances vol 9:260), by the process of liquid outside blood to tumour cell, and cancer cell
Interior endocytosis body and the destructive process of lysosome, it is that weakly acidic process is changed into from weakly alkaline environment, drug molecule is by molecule
State is changed into ionic state, and the rise of its solubility, i.e., fat-soluble to be changed into hydrophily, being discharged from surface of graphene oxide increases, and is dissolved in
In intracellular fluid, combined with DNA topoisomerases, this is to be played by principle sensitive pH by the use of graphene oxide as carrier
The toxic treatment effect of medicine.
Found in tumor research, disulfide bond (- S-S-) is stabilized under extracellular temperate condition, and is run into the cell
Reducing substances is decomposed rapidly by disulfide bond exchange reaction, these reducing substances be mainly glutathione (glutathione,
GSH, Methods Enzymol, 1995,251:8-28；Annu Rev Microbiol,1997,51:179-202).GSH is thin
(about 0.5~10mM) be present in cytosol and subcellular fraction core with millimolar concentration high level, yet with the rapid enzyme in blood plasma
Fall below reduced levels (about 2~20 μm), i.e. cytoplasm and endonuclear reduction potential is about body fluid or extracellular
The body fluid such as liquid 100~1000 times (Cell Biochem Funct, 2004,22:343-352；Free Radic Biol Med,
2001,30:1191-1212).Tumour cell is fast due to growing, and oxygen demand is larger, causes GSH content to be at least normal structure
4 times of (Cancer Res, 2002,62 (1):307-312), therefore, cancer therapy drug or gene are passed using redox principle
It is delivered in tumour cell and plays therapeutic action.
Heparin (heparin, seeing Fig. 3) is being made up of the glycuronic acid and gucosamine disaccharides of 1 → 4 connection for discovery in 1916
Linear polysaccharide class compound, nineteen thirty-five clinically starts to be used for anti-coagulants, its activity be because it can and serine protease
Inhibitor antithrombase is combined, cause the inhibitor become inactive fibrin ferment (J Med Chem, 2003,46:2551-
64).What common heparin typically extracted from the natural tissues such as chitterlings or ox lung, mean molecule quantity about 15kDa, about
Have 20 disaccharide structures, chemical constitution and molecular weight are highly non-uniform, polydispersity be 1.2~1.4, molecular weight ranges be 5~
40kDa.Sodium salt is readily soluble in water, the indissoluble in it common are solvent, and strand stretches curl in water, and 12 double
The heparin length of sugared unit is about 5nm, thus it is speculated that the length of heparin is about 9nm.Each disaccharide unit contains a carboxyl, one or
Multiple 1 ° or 2 ° of hydroxyls, 2~2.5 sulfates, wherein N- sulfuric acid fiduciary point 75~85%, O- sulfuric acid fiduciary point 15~25%, each
Also containing 15~25% adjacent hydroxyl in disaccharide unit structure.Each chain contains the reducing end under neutral of a hemiacetal, masks
Aldehyde radical (CHO).The each chain of heparin contains about 0.3 free amino (NH2).Highly-hydrophilic, even if in dry conditions, still carrying
2~10% water.Because containing carboxyl (pKa 3.3) and sulfate, (pKa of O- sulfates and N- sulfates is 1.0 in structure
~1.5), make heparin highly negatively charged (about -75/ chain) so that can be with more hatching eggs such as growth factor, protease, cell factor
White matter occur electrostatic interaction (Nat Prod Rep, 2002,19:312-331), cause protein stabilization or add to cell by
Body compatibility (J Cell Physiol, 1986,128:475-484), this stability has been used to fibroblastic growth
The factor (fibroblast growth factor, FGF) and VEGF (vascular endothelial
Growth factor, VEGF) it can be made into regeneration and put down with the drug delivery system of engineering carrier bracket and Drug controlled release
Platform (Chem Soc Rev, 2013,42:7335-7372).As anti-coagulants, the more side effect of heparin, as hemorrhagic is concurrent
After disease, decrease of platelet, non-vein administration bioavilability it is low (J Clin Pharmacol, 1992,32:584-596).
LMWHs (low-molecular weight heparin, LMWH) chemical composition is definite, biological half-life
Extend, adverse reaction it is less (Science, 2011,334:498-501).LMWHs (CAS:9041-08-1) there are a variety of medicines
With form, such as medicinal low molecular weight heparin sodium, low molecular weight calcium heparin, Ardeparin Sodium (Ardeparin Sodium), shellfish
Rice liquaemin (Bemiparin Sodium), Certoparin Sodium (Certoparin Sodium), Dalteparin Sodium (Dalteparin
Sodium), Enoxaparin Sodium (Enoxaparin Sodium), nadroparin calcium (Nadroparin Calcium), Parnaparin Sodium
(Parnaparin Sodium), Clivarin sodium (Reviparin Sodium), tinzaparin sodium (Tinzaparin
Sodium) etc. (Martindale, 36 editions page 1329).For studies have shown that compared with common unassorted heparin, LMWH passes through regulation
The growth factor (such as FGF and VEGF) related to a variety of blood vessels, the more effective fruit of growth to suppressing tumor tissues.In addition, LMWH
May also interfere with metastases, or with P-selectin competitive binding, suppress tumour it is small stick (Nat Rev
Cancer,2002,2:521-528).Recently research is found, heparin can also act on transcription factor, cause apoptotic death
(Chem Biol,2004,11:420-422).Therefore, LMWH is widely used in Tumor Targeting Drug Delivery System, also there is some chemistry
The LMWH derivatives of modification, such as LMWH- deoxycholic acids, anticoagulating active reduces, anti-angiogenesis activity rise (J Controlled
Release,2010,148:317-326).LMWH can be used as antifibrotic agents treatment hepatitis B (Biomaterials, 2011,32:
1438-1445).Heparin modified PLGA- pluronics F-127 can be used for control growth factor release (Biomaterials,
2006,27:2621-2626), Kipper et al. utilizes polyanion chitosan-heparin and polycation Chitosan-Hyaluronic Acid
Polymer electrolyte verify composition compound polyelectrolyte nanoparticle (polyelectrolyte complex nanoparticles,
PCN), the release for controlled release FGF-2, can slowly be discharged in a manner of Zero order release 30 days (Acta Biomater, 2012,8:
1551-1559).The nanoparticle formed with chlorination trimethyl chitin and heparin non-covalent bond, it is prominent to release using VEGF as model drug
Effect is reduced, and 14 days zero-order fashions release about 49% (Macromol Rapid Commun, 2012,33:2015-2022).Mi etc.
People has synthesized heparin modified chitosan/Polyurethane-epoxy resin nanoparticle, because gamma-glutamic acid and heparin are negatively charged, chitosan band
On schedule, material is sensitive to pH, stable under the conditions of pH 6.0, gradually expands under the conditions of pH 6.6 but is not disintegrated, but in pH
Be disintegrated rapidly under the conditions of 7.4, this be due to the nanoparticle lose electrostatic interaction (Biomaterials, 2010,31:9320-
9332).Using heparin and bFGF as model, under the slightly sour environment of ischemic tissue, nanoparticle keeps complete, delays Slow release,
In repair tissue (pH7.4), heparin can be discharged rapidly, and these characteristics can suppress vascular thrombosis and re-form.BFGF release can show
Work makes human foreskin fibroblasts propagation increase, and is effectively facilitated Human umbilical vein endothelial cells segment dislocation.
Although graphene oxide itself has more oxy radical (such as hydroxyl, epoxy radicals and carboxyl), Fig. 4 is graphene oxide
Typical IR collection of illustrative plates, but because precursor structure has substantial amounts of carbon-carbon double bond, therefore, water solubility is not especially good, such as aoxidizes stone
For the black usual concentration of alkene when more than 5mg/mL, long-term place easily forms g., jelly-like material；When concentration is more than 20 μ g/mL, oxygen
Graphite alkene starts to be gathered into lamellar structure, when concentration is 50 μ g/mL, concentration class reach 15% (Langmuir, 2013,29:
13443-13448).In inorganic salt solution (such as sodium chloride, sodium phosphate), graphene oxide can self-assemble into lamella knot
Structure, and precipitation is produced, therefore, if as drug carrier material, it need to be modified water-soluble to improve it.Aoxidized for reproducibility
Graphene, Fig. 5 is the typical IR collection of illustrative plates of reproducibility graphene oxide, and because oxy radical is largely reduced, therefore, water solubility is more
Difference, it is water-soluble that it need to be improved with more hydrophilic radical.Research finds that heparin is a kind of good water soluble molecules (1 part of liver
Plain sodium is dissolved in 20 parts of water, Martindale, 36 editions:Page 1301), it plays anticancer using itself there is blood vessel formation against function
Activity.It is administered with together with cancer therapy drug, there is synergistic therapeutic action.At present, graphene oxide is combined, simultaneously with heparin or its salt
Apply on drug carrier material there is not yet relevant report.
The content of the invention
An object of the present invention is to provide a kind of heparin or the graphene oxide of its salt modification and preparation method thereof with answering
With the graphene oxide after heparin or the modification of its salt can be as the carrier material of medicine particularly cancer therapy drug.
A kind of heparin or the graphene oxide of its salt modification, are prepared one of by the following method：
(1) according to pH sensitivity principles, using adipic dihydrazide as connection molecule, between heparin or its salt and graphene oxide
It is connected by amido link and obtains the graphene oxide that heparin or its salt are modified；
(2) according to redox principle, using cystamine as connection molecule, two are passed through between heparin or its salt and graphene oxide
Sulfide linkage, which is connected, obtains the graphene oxide that heparin or its salt are modified.
In method (1), comprise the following steps：Heparin or its salt are modified using adipic dihydrazide, obtain the acyl of adipic acid two
Heparin or its salt after hydrazine modification, heparin or its salt after then obtained adipic dihydrazide is modified and the oxidation after activation
Graphene reacts 6~24h at 25~30 DEG C, obtains the oxygen modified using adipic dihydrazide as the heparin of connection molecule or its salt
Graphite alkene stops polymer solution.
Wherein, the method for the adipic dihydrazide modification heparin or its salt is：Liquaemin is soluble in water, adds adipic acid
Two hydrazides, 1- ethyls-(3- dimethylaminopropyls) phosphinylidyne diimmonium salt hydrochlorate (EDCHCl) and I-hydroxybenzotriazole
(HOBt) solution ph, is adjusted to 6.0~7.0.12~48h of stirring reaction at 25~30 DEG C.Unreacted starting material is used
Dialysis removes, and dries, and obtains the heparin (product A) after the modification of product adipic dihydrazide.Described liquaemin and mole of water
Than for 1:(10000~100000)；Described liquaemin and the mol ratio of adipic dihydrazide are 1:(2~60)；Described liver
The mol ratio of plain sodium and EDCHCl is 1:(2~60)；Described liquaemin and HOBt mol ratio are 1:(2~60).This hair
The connection effect of the method for adipic dihydrazide modification heparin or its salt in bright, adipic dihydrazide modification heparin or its salt
It is good, improve the yield of product in end reaction.But adipic dihydrazide-heparin of the present invention or the preparation method of its salt are not only
It is limited to the method.
The preparation method of graphene oxide after activation is：Graphene oxide solution is taken, adds EDCHCl and N- hydroxyls
Succinimide sulfonate sodium (sulfur-NHS), regulation solution ph to 6.0~7.0, at 25~30 DEG C stirring reaction 1~
3 hours, the graphene oxide after being activated.The method of active oxidation graphene in the present invention is by surface of graphene oxide official
Energy group carries out priming reaction so that activation effect is preferable, improves the yield of product in end reaction.But the activation oxygen of the present invention
The preparation method of graphite alkene (GO) is not limited only to the method.
In method (2), comprise the following steps：Heparin or its salt are modified using cystamine, obtain the heparin after cystamine modification or
Its salt, then by obtained cystamine modify after heparin or its salt with activation after graphene oxide 25~30 DEG C react 6~
24h, the graphene oxide for obtaining modifying using cystamine as the heparin of connection molecule or its salt stop polymer solution.
Wherein, the method for the cystamine modification heparin or its salt is：Liquaemin is soluble in water, adds cystamine (Cys),
EDCHCl and HOBt, regulation solution ph to 6.0~7.0.12~48h of stirring reaction at 25~30 DEG C.Unreacted
Beginning material is removed with dialysis, is dried, and obtains the heparin (product B) after the modification of product cystamine.Described liquaemin and mole of water
Than for 1:(10000~100000)；Described liquaemin and the mol ratio of cystamine are 1:(2~60)；Described liquaemin with
EDCHCl mol ratio is 1:(2~60)；Described liquaemin and HOBt mol ratio are 1:(2~60).In the present invention
The method that cystamine modifies heparin, cystamine modify the good connecting effect of heparin, improve the yield of product in end reaction.But the present invention
The preparation method of cystamine-heparin be not limited only to the method.
The preparation method of graphene oxide after activation is：Graphene oxide solution is taken, adds EDCHCl and sulfur-
NHS, regulation solution ph is to 6.0~7.0, stirring reaction 1~3 hour at 25~30 DEG C.
Described graphene oxide solution concentration is 0.5~1.5mg/mL；The weight of described graphene oxide and product B
Than for 1:(2~20)；Described product B and EDCHCl weight ratio is 1:(1~5)；Described product A and sulfur-NHS
Weight ratio be 1:(1~5).
It is a further object of the present invention to provide a kind of anticancer compound, the anticancer compound is to be prepared by the following method
Arrive：In the graphene oxide that the heparin or its salt are modified, cancer therapy drug (Drug) solution is added, is stirred, dialysis, is removed not
The medicine contained, obtains anticancer drug complex.The principle that the graphene oxide that medicine is modified with heparin or its salt is combined is thing
Reason absorption, forms compound.
Heretofore described graphene oxide refers to graphene oxide or the reproducibility oxidation stone after reducing agent reduces
Black alkene.The present invention can use the graphene oxide of two kinds of forms, and efficiency high good with the reaction effect of heparin or its salt.
In the present invention, it is preferred that the preparation method of the graphene oxide (GO) is：Graphite powder is in the concentrated sulfuric acid, persulfuric acid
In the presence of potassium and phosphorus pentoxide, the graphite powder (or graphite powder for expansion) of pre-oxidation is obtained.The graphite powder of pre-oxidation is dense
Under sulfuric acid and potassium permanganate effect, graphene oxide is generated.After the aqueous terminating reaction for adding hydrogen peroxide, graphene oxide is obtained
Precipitation, then precipitated respectively with dilute hydrochloric acid solution, water washing, and dialysed with purified water, ultrasound, obtain the molten of graphene oxide
Liquid.The graphene oxide obtained using the method, effect are preferable, there is provided the good basis of connection heparin or its molecules of salt.But this
The preparation method of the graphene oxide (GO) of invention is not limited only to the method.
The preparation method of the reproducibility graphene oxide (rGO) is：The GO colloidal solution is taken, vitamin C is added, adds
It is ultrasonic under heat condition, sodium chloride is added, is stood, centrifugation, collecting to precipitate, and respectively with respectively washing 2 times of ethanol and pure water, centrifuge
Reproducibility graphene oxide, and with purifying aqueous suspension, ultrasound, obtained rGO solution.The reproducibility oxygen obtained using the method
Graphite alkene, effect are preferable, there is provided the good basis of connection heparin or its molecules of salt.But the reproducibility graphene oxide of the present invention
Preparation method be not limited only to the method.
Liquaemin of the present invention refers to the one or two kinds of combination therein of unfractionated heparin sodium, low molecular sodium heparin；
The medicinal forms of the low molecular sodium heparin are medicinal low molecular sodium heparin, Low-molecular-weight Heparins Calcium, Certoparin Sodium, shellfish rice liver
Plain sodium, Dalteparin Sodium, Enoxaparin Sodium, nadroparin calcium, Parnaparin Sodium, Clivarin sodium, tinzaparin sodium, Ardeparin Sodium
One or more kinds of combinations.Using the liquaemin of above form, and efficiency high good with the reaction effect of graphene oxide.
It is as follows according to the sensitive principles of pH, specific preparation method：
A：Graphene oxide-heparin stops compound/cancer therapy drug
(1) graphene oxide (GO) prepares graphite powder in the presence of the concentrated sulfuric acid, potassium peroxydisulfate and phosphorus pentoxide, obtains
The graphite powder (or graphite powder for expansion) of pre-oxidation.Under the graphite powder concentrated sulfuric acid and the potassium permanganate effect of pre-oxidation, generation
Graphene oxide.After the aqueous terminating reaction for adding hydrogen peroxide, the precipitation of graphene oxide is obtained, then it is molten with watery hydrochloric acid respectively
Liquid, water washing precipitation, and dialysed with purified water, ultrasound, obtain the colloidal solution of graphene oxide.
The heparin (ADH-UFH) of adipic dihydrazide modification to prepare liquaemin soluble in water, add the acyl of adipic acid two
Hydrazine, 1- ethyls-(3- dimethylaminopropyls) phosphinylidyne diimmonium salt hydrochlorate (EDCHCl) and I-hydroxybenzotriazole (HOBt),
Solution ph is adjusted to 6.0~7.0.12~48h of stirring reaction at 25~30 DEG C.Unreacted starting material purified water is saturating
Analysis removes, and freeze-drying, obtains product ADH-UFH.Described liquaemin and the mol ratio of water are 1:(10000~100000)；
Described liquaemin and the mol ratio of adipic dihydrazide are 1:(2~60)；Described liquaemin and EDCHCl mol ratio
For 1:(2~60)；Described liquaemin and HOBt mol ratio are 1:(2~60).
(3) the GO (GO-ADH-UFH) of ADH-UFH modifications preparation takes graphene oxide solution, adds EDCHCl and N-
HOSu NHS sulfonate sodium (sulfur-NHS), regulation solution ph stir anti-to 6.0~7.0 at 25~30 DEG C
Answer 1~3 hour.ADH-UFH, stirring reaction 6~24 hours at 25~30 DEG C are added, dialysis removes unreacted EDC
HCl, sulfur-NHS and ADH-UFH, obtain GO-ADH-UFH and stop polymer solution.
The colloidal solution concentration of described graphene oxide is 0.5~1.5mg/mL；Described graphene oxide and product
ADH-UFH weight ratio is 1:(2~20)；Described product ADH-UFH and EDCHCl weight ratio is 1:(1~5)；It is described
ADH-UFH and sulfur-NHS weight ratio is 1:(1~5).
(4) the preparation containing anticancer drug complex stops polymer solution in GO-ADH-UFH, and it is molten to add cancer therapy drug (Drug)
Liquid, stir, dialysis, remove the medicine of unentrapped, obtain anticancer drug complex (GO-ADH-UFH/Drug).
B：Reproducibility graphene oxide-heparin stops compound/cancer therapy drug
The preparation of graphene oxide with above-mentioned A (1).
Adipic dihydrazide modification heparin preparation with above-mentioned A (2).
(3) the preparation of reproducibility graphene oxide (rGO) takes GO colloidal solution, adds vitamin C, surpasses under heating condition
Sound, sodium chloride is added, stood, centrifugation, collecting to precipitate, and respectively with respectively washing 2 times of ethanol and pure water, centrifuge to obtain reproducibility oxidation
Graphene, and with purifying aqueous suspension, ultrasound, obtained rGO colloidal solution.
(4) the rGO (rGO-ADH-UFH) of ADH-UFH modifications preparations takes rGO colloidal solution, add EDCHCl with
Sulfur-NHS, regulation solution ph is to 6.0~7.0, stirring reaction 1~3 hour at 25~30 DEG C.ADH-UFH is added,
Stirring reaction 6~24 hours at 25~30 DEG C, dialysis remove unreacted EDCHCl, sulfur-NHS and ADH-UFH, obtained
GO-ADH-UFH stops polymer solution.
The colloidal solution concentration of the rGO is 0.5~1.5mg/mL；The weight ratio of the rGO and ADH-UFH are 1:(2~
20)；Described ADH-UFH and EDCHCl weight ratio are 1:(1~5)；Described ADH-UFH and sulfur-NHS weight
Than for 1:(1~5).
(5) the preparation containing anticancer drug complex stops polymer solution in rGO-ADH-UFH, and it is molten to add cancer therapy drug (Drug)
Liquid, stir, dialysis, remove the medicine of unentrapped, obtain anticancer drug complex (rGO-ADH-UFH/Drug).
C：Graphene oxide-LMWHs stops compound/cancer therapy drug
The preparation of graphene oxide (GO) with above-mentioned A (1).
The LMWHs (ADH-LMWH) of adipic dihydrazide modification to prepare low molecular sodium heparin soluble in water, add
Enter adipic dihydrazide (ADH), 1- ethyls-(3- dimethylaminopropyls) phosphinylidyne diimmonium salt hydrochlorate (EDCHCl) and 1- hydroxyls
Base BTA (HOBt), the value of solution is adjusted to 6.0~7.0.12~48h of stirring reaction at 25~30 DEG C.Unreacted reactant
Dialysed and removed with water, freeze-drying, obtain product ADH-LMWH.
Described low molecular sodium heparin and the mol ratio of water are 1:(10000~100000)；Described liquaemin with oneself two
The mol ratio of acid dihydrazide is 1:(2~60)；Described liquaemin and EDCHCl mol ratio are 1:(2~60)；Described
Liquaemin and HOBt mol ratio are 1:(2~60).
(3) the GO (GO-ADH-LMWH) of ADH-LMWH modifications preparation takes graphene oxide solution, add EDCHCl and
N-hydroxysuccinimide sulfonate sodium (sulfur-NHS), regulation solution value stir anti-to 6.0~7.0 at 25~30 DEG C
Answer 1~3 hour.ADH-LMWH, stirring reaction 6~24 hours at 25~30 DEG C are added, dialysis removes unreacted EDC
HCl, sulfur-NHS and ADH-LMWH, obtain GO-ADH-LMWH and stop polymer solution.
Described graphene oxide solution concentration is 0.5~1.5mg/mL；Described graphene oxide and ADH-LMWH's
Weight ratio is 1:(2~20)；The weight ratio of the ADH-LMWH and EDCHCl are 1:(1~5)；The ADH-LMWH with
Sulfur-NHS weight ratio is 1:(1~5).
(4) the preparation containing anticancer drug complex stops polymer solution in GO-ADH-LMWH, and it is molten to add cancer therapy drug (Drug)
Liquid, stir, dialysis, remove the medicine of unentrapped, obtain anticancer drug complex (GO-ADH-LMWH/Drug).
D：Reproducibility graphene oxide-LMWHs stops compound/cancer therapy drug
The preparation of graphene oxide with above-mentioned A (1).
Adipic dihydrazide modification LMWHs preparation with above-mentioned C (2).
The preparation of reproducibility graphene oxide (rGO) with above-mentioned B (3).
(4) the rGO (rGO-ADH-LMWH) of ADH-LMWH modifications preparation takes rGO colloidal solution, adds EDCHCl
And sulfur-NHS, regulation solution ph is to 6.0~7.0, stirring reaction 1~3 hour at 25~30 DEG C.Add ADH-
LMWH, stirring reaction 6~24 hours at 25~30 DEG C, dialysis remove unreacted EDCHCl, sulfur-NHS and ADH-
LMWH, obtain GO-ADH-LMWH and stop polymer solution.
The reproducibility graphene oxide colloidal solution concentration is 0.5~1.5mg/mL；The reproducibility graphene oxide
Weight ratio with ADH-LMWH is 1:(2~20)；The weight ratio of the ADH-LMWH and EDCHCl are 1:(1~5)；It is described
ADH-LMWH and sulfur-NHS weight ratio is 1:(1~5).
(5) the preparation containing anticancer drug complex stops polymer solution in rGO-ADH-UFH, and it is molten to add cancer therapy drug (Drug)
Liquid, stir, dialysis, remove the medicine of unentrapped, obtain anticancer drug complex (rGO-ADH-UFH/Drug).
Using pH sensitivity principles, a kind of heparin modified graphene oxide, the graphene oxide refers to graphite oxide
Alkene or the reproducibility graphene oxide after reducing agent reduces；
Using pH sensitivity principles, a kind of heparin modified graphene oxide, the liquaemin refer to unfractionated heparin sodium or
Low molecular sodium heparin；
Using pH sensitivity principles, a kind of heparin modified graphene oxide be using adipic dihydrazide as connection molecule and
The graphene oxide of unfractionated heparin modification.
Using pH sensitivity principles, a kind of heparin modified graphene oxide be using adipic dihydrazide as connection molecule and
The reproducibility graphene oxide of unfractionated heparin modification.
Using pH sensitivity principles, a kind of heparin modified graphene oxide be using adipic dihydrazide as connection molecule and
The graphene oxide of LMWHs modification.
Using pH sensitivity principles, a kind of heparin modified graphene oxide be using adipic dihydrazide as connection molecule and
The reproducibility graphene oxide of LMWHs modification.
According to redox principle, specific preparation method is as follows：
E：Graphene oxide-heparin stops compound/cancer therapy drug
The preparation of graphene oxide (GO) with above-mentioned A (1).
The heparin (Cys-UFH) of cystamine modification to prepare liquaemin soluble in water, add cystamine (Cys), EDCHCl
And HOBt, the pH value of solution is adjusted to 6.0~7.0.12~48h of stirring reaction at 25~30 DEG C.Unreacted reactant is dialysed with water
Remove, freeze-drying, obtain product Cys-UFH.
Described liquaemin and the mol ratio of water are 1:(10000~100000)；Described liquaemin and mole of cystamine
Than for 1:(2~60)；Described liquaemin and EDCHCl mol ratio are 1:(2~60)；Described liquaemin and HOBt's
Mol ratio is 1:(2~60)
(3) the GO (GO-ss-UFH) of Cys-UFH modifications preparation takes graphene oxide solution, add EDCHCl and
Sulfur-NHS, regulation solution ph is to 6.0~7.0, stirring reaction 1~3 hour at 25~30 DEG C.Cys-UFH is added, is stirred
Mix overnight, dialysis removes unreacted EDCHCl, sulfur-NHS and Cys-UFH, obtains GO-ss-UFH and stops polymer solution.
The graphene oxide colloidal solution concentration is 0.5~1.5mg/mL；The weight of the graphene oxide and Cys-UFH
Amount is than being 1:(2~20)；The weight ratio of the Cys-UFH and EDCHCl are 1:(1~5)；The Cys-UFH-LMWH with
Sulfur-NHS weight ratio is 1:(1~5).
(4) the preparation containing anticancer drug complex stops polymer solution in GO-ss-UFH, and it is molten to add cancer therapy drug (Drug)
Liquid, stir, dialysis, remove the medicine of unentrapped, obtain anticancer drug complex (GO-ss-UFH/Drug).
F：Reproducibility graphene oxide-heparin stops compound/cancer therapy drug
The preparation of graphene oxide with above-mentioned A (1).
Cystamine modification heparin (Cys-UFH) preparation with above-mentioned E (2).
(3) the preparation of reproducibility graphene oxide (rGO) takes GO colloidal solution, adds vitamin C, surpasses under heating condition
Sound, sodium chloride is added, stood, centrifugation, collecting to precipitate, and respectively with respectively washing 2 times of ethanol and pure water, centrifuge to obtain reproducibility oxidation
Graphene, and with purifying aqueous suspension, ultrasound, obtained rGO colloidal solution.
(4) the rGO (rGO-ss-UFH) of Cys-UFH modifications preparation takes rGO solution, adds cystamine (Cys), EDCHCl
And HOBt, the pH value of solution is adjusted to 6.0~7.0, stirring reaction 1~3 hour at 25~30 DEG C.Unreacted reactant is dialysed with water
Remove, freeze-drying, obtain product.
The rGO colloidal solution concentration is 0.5~1.5mg/mL；Described rGO and product B weight ratio is 1:(2~
20)；The weight ratio of the Cys-UFH and EDCHCl are 1:(1~5)；Described product Cys-UFH and sulfur-NHS weight
Amount is than being 1:(1~5).
(5) the preparation containing anticancer drug complex stops polymer solution in rGO-ss-UFH, and it is molten to add cancer therapy drug (Drug)
Liquid, stir, dialysis, remove the medicine of unentrapped, obtain anticancer drug complex (rGO-ss-UFH/Drug).
G：Graphene oxide-LMWHs stops compound/cancer therapy drug
The preparation of graphene oxide (GO) with above-mentioned A (1).
The LMWHs (Cys-LMWH) of cystamine modification to prepare low molecular sodium heparin soluble in water, add oneself two
Acid dihydrazide (ADH), EDCHCl and HOBt, the pH value of solution is adjusted to 6.0~7.0.The stirring reaction 12 at 25~30 DEG C
~48h.Unreacted reactant is dialysed with water dialysis and removed, and freeze-drying, obtains product.
Described low molecular sodium heparin and the mol ratio of water are 1:(10000~100000)；The low molecular sodium heparin with
The mol ratio of cystamine is 1:(2~60)；Described low molecular sodium heparin and EDCHCl mol ratio are 1:(2~60)；It is described
Low molecular sodium heparin and HOBt mol ratio be 1:(2~60).
(3) the GO (GO-ss-LMWH) of Cys-LMWH modifications preparation takes graphene oxide solution, add EDCHCl and
Sulfur-NHS, regulation solution value is to 6.0~7.0, stirring reaction 1~3 hour at 25~30 DEG C.Cys-LMWH is added,
Stirring reaction 6~24 hours at 25~30 DEG C, dialysis remove unreacted EDCHCl, sulfur-NHS and Cys-LMWH, obtained
Stopped polymer solution to GO-ss-LMWH.
Described graphene oxide solution concentration is 0.5~1.5mg/mL；Described graphene oxide and Cys-UFH weight
Amount is than being 1:(2~20)；The weight ratio of the Cys-UFH and EDCHCl are 1:(1~5)；The Cys-UFH and sulfur-
NHS weight ratio is 1:(1~5).
(4) the preparation containing anticancer drug complex stops polymer solution in GO-ss-LMWH, and it is molten to add cancer therapy drug (Drug)
Liquid, stir, dialysis, remove the medicine of unentrapped, obtain anticancer drug complex (GO-ss-LMWH/Drug).
H：Reproducibility graphene oxide-heparin stops compound/cancer therapy drug
The preparation of graphene oxide with above-mentioned A (1).
Cystamine modification heparin preparation with above-mentioned G (2).
The preparation of reproducibility graphene oxide (rGO) with above-mentioned B (3).
(4) the rGO (rGO-ss-LMWH) of Cys-LMWH modifications preparations takes rGO colloidal solution, add EDCHCl with
Sulfur-NHS, regulation solution ph is to 6.0~7.0,1~3h of stirring reaction at 25~30 DEG C.Cys-LMWH is added, 25
Stirring reaction 6~24 hours at~30 DEG C, dialysis remove unreacted EDCHCl, sulfur-NHS and Cys-LMWH, obtained
GO-ss-LMWH stops polymer solution.
Described rGO colloidal solution concentration is 0.5~1.5mg/mL；The weight ratio of the rGO and Cys-LMWH are 1:
(2~20)；The weight ratio of the Cys-LMWH and EDCHCl are 1:(1~5)；The weight of the Cys-LMWH and sulfur-NHS
Amount is than being 1:(1~5).
(5) the preparation containing anticancer drug complex stops polymer solution in rGO-ADH-UFH, and it is molten to add cancer therapy drug (Drug)
Liquid, stir, dialysis, remove the medicine of unentrapped, obtain anticancer drug complex (rGO-ss-UFH/Drug).
Using redox principle, a kind of heparin modified graphene oxide, the graphene oxide refers to aoxidizing stone
Black alkene or the reproducibility graphene oxide after reducing agent reduces.
Using redox principle, a kind of heparin modified graphene oxide, the liquaemin refers to unfractionated heparin sodium
Or low molecular sodium heparin.
Using redox principle, a kind of heparin modified graphene oxide is to be used as connection molecule and common liver using cystamine
The graphene oxide of element modification.
Using redox principle, a kind of heparin modified graphene oxide is to be used as connection molecule and common liver using cystamine
The reproducibility graphene oxide of element modification.
Using redox principle, a kind of heparin modified graphene oxide is to be used as connection molecule and low molecule using cystamine
Heparin modified graphene oxide.
Using redox principle, a kind of heparin modified graphene oxide is to be used as connection molecule and low molecule using cystamine
Heparin modified reproducibility graphene oxide.
According to quality standard WS1- (X-149) -2005Z, low molecular sodium heparin are to be refined with liquaemin through nitrous acid cleavage
Obtained by CSSO3 sodium salt (new drug become a full member the 66th the 193-196 pages of standard).
According to quality standard WS1- (X-147) -2005Z, Low-molecular-weight Heparins Calcium are to be refined with liquaemin through nitrous acid cleavage
Obtained by CSSO3 calcium salt (new drug become a full member the 66th the 176-179 pages of standard).Low molecular sodium heparin and low point
The weight average molecular weight of sub- calciparine is respectively less than 8000, and component of the molecular weight less than 8000 is no less than the 60% of total amount.
Certoparin Sodium is produced by Aspen Pharma companies, and trade name has：Root
Obtained according to nitrous depolymerisation method, its mean molecule quantity is 6000, and its 70% component molecules amount is not more than 10000
(Martindale, 36 editions page 1242).
Bemiparin is produced by Rovi drugmakers, and trade name has：With), mean molecule quantity 3000-4200, its characteristic molecular amount is 3600, and about 85% molecular weight is less than 6000, two
Sugar unit contains about 2.0 sulfuric acid unit (Drugs.2003,63 (21):2357-2377；Martindale, 36 editions page 1223).
Dalteparin Sodium is produced and sold by Pfizer, and trade name has：), according to nitrous acidolysis
What poly- method obtained, its mean molecule quantity is 5600-6400, and its characteristic molecular amount is 6000, less than 3000 molecular weight component not
More than 13%, more than 8000 molecular weight component in the range of 15-25%, disaccharide unit contains 2.0-2.5 sulfuric acid unit (Europe
7.0 editions the 1788-1789 pages of continent pharmacopeia；Martindale, 36 editions page 1255).
Enoxaparin Sodium is produced and sold by matching Norfin, Inc, and trade name has：
With), obtained according to the Benzylation rear poly- method of alkaline hydrolysis that carries out, its mean molecule quantity is 3800-5000, and its feature is divided
Son amount is 4500 (7.0 editions the 1920-1921 pages of European Pharmacopoeias).
Nadroparin calcium is produced and sold by matching Norfin, Inc and GSK companies, and trade name has：Produced and sold by Noble Molecules companies, trade name：Using nitrous
Acid depolymerization method obtains, and its mean molecule quantity is 3600-5000, and its characteristic molecular amount is 4300, less than the group of 2000 molecular weight
Divide and be not more than 15% (7.0 editions the 2543-2545 pages of European Pharmacopoeia).
Parnaparin Sodium is produced and sold by Italian Alfa Wassermann drugmakers, and trade name has：Japanese Japan's pharmaceutical industries company produces and sells, trade name：Japanese aginomoto system
Medicine company trade name：Obtained according to peroxidative depolymerization method, its mean molecule quantity is 4000-6000, and it is special
It is 5000 to levy molecular weight, and the component less than 3000 molecular weight is not more than 30% (European Pharmacopoeia 7.0 edition page 2672).
Clivarin sodium is produced and sold by German Knoll AG companies, and trade name has：Adopt
Obtained with nitrous depolymerisation method, its mean molecule quantity is 3900 (Expert Opin Pharmacother.2002,3 (2):
173-182) also have been reported that, its mean molecule quantity is 3150-5150, contains 2.1 sulfate groups in its disaccharide unit
(Martindale, 36 editions:Page 1388).
Tinzaparin sodium is produced and sold by Aktiebolaget Leo (SE) Box 941, S-251 09 Helsingborg, Sweden of Denmark, trade name：Produced and sold by Novo companies
Sell trade name：β-elimination depolymerization being carried out using heparinase to obtain, its mean molecule quantity is 5500-7500, its
Characteristic molecular amount is 6500, and the component less than 2000 molecular weight is not more than 10% (7.0 editions the 3098-3099 pages of European Pharmacopoeia).
Ardeparin Sodium is produced by Wyeth Pharmaceuticals, trade name：Normiflo, obtained according to peroxidative depolymerization, its
For 98% component molecules amount in 2000-15000, its mean molecule quantity is 5500-6500, contains 2.7 sulphur in its disaccharide unit
Acid groups (Martindale, 36 editions page 1216), but the reason for be not due to safety or validity and the reason for be due to regulation
City (J Thromb Thrombolysis.2001,11 (3) are removed from American market within 2000:247-259；Carbohydr
Polym.2013,97(2):684-689)。
In the present invention, the mean molecule quantity of unfractionated heparin sodium (UFH) calculates with 1.5 ten thousand, and the average mark of LMWHs
Son amount calculates with 0.5 ten thousand, is fed intake with mol ratio.Graphene oxide or reproducibility graphene oxide are due to laminated structure size
Differ, itself is without the molecular weight or average molecular weight range determined, separately because ingredient proportion is different, the obtained acyl of adipic acid two
The graphene oxide or reproducibility graphene oxide substitution value of the heparin and final chemical modification of hydrazine or cystamine chemical modification are not
Together, therefore in the present invention, heparin and graphene oxide or reproducibility oxygen containing adipic dihydrazide or cystamine chemical modification
The material of graphite alkene is fed intake with weight ratio.
The beneficial effects of the invention are as follows：
(1) containing the oxy radical such as substantial amounts of hydroxyl, epoxy radicals, carboxyl in graphene oxide self structure, and with compared with
Good hydrophily, can be widely applied in biomedicine.Because graphene oxide is monoatomic layer structure, there is larger ratio table
Area (has document to be measured as 736.6m2/ g, is shown in Langmuir, 2013,29:13443-13448), its two sides all can be by altogether
Valency, noncovalent interaction are combined with medicine, therefore have higher drug load amount.It can by stronger physisorption with
Aromatic rings class medicine Non-covalent binding, some insoluble drugs, the particularly body to most of slightly solubility cancer therapy drug can be delivered
Interior transhipment has great importance.Heparin is a kind of good water soluble molecules, its using itself there is blood vessel formation against function and
Play active anticancer.It is administered with together with cancer therapy drug, there is synergistic therapeutic action.The present invention is existed using graphene oxide and heparin
The advantage of drug field, root it was found that being attached heparin and graphene oxide, obtain it is heparin modified after graphite oxide
Alkene.
(2) present invention is according to two kinds of principle research heparin and the connectivity problem of graphene oxide, and the first scheme is according to pH
Sensitive principle.I.e. using in tumour cell and extracellular fluid has relatively low pH, graphene oxide or reproducibility graphite oxide
The carboxyl (COOH) of alkene and active group (O=C-NH-NH in adipic dihydrazide (ADH) structure2) form biamide structure (O
=C-NH-NH-C=O), it is prepared into the sensitive compounds that stop (conjugates) of pH.Meta-alkalescence blood (pH 7.35~
7.45) in, the biamide structure can not hydrolyze, and heparin is located at the both sides of graphene oxide, can increase the water-soluble of graphene oxide
Property, while its pi-pi accumulation and hydrophobic effect are utilized, antineoplastic in physical absorption, form compound (complexs)；When this
Compound transports target cell (such as cancer cell) place, using strengthen penetrating and retention effect (see Lu Bin edit novel pharmaceutical formulations with
New technology, the second edition, page 63), enter in target cell, in the slant acidity microenvironment of target cell, compound discharges anti-
Cancer drug, simultaneously as biamide structure (O=C-NH-NH-C=O) fracture in the compound that stops, forms heparin and oxidation stone
Black alkene, the former has blood vessel formation against function, and synergistic therapeutic action is played to anticancer.
Second scheme is according to redox principle.It is i.e. stronger using reproducibility in tumour cell, contain higher concentration
Glutathione (GSH), the compound for containing (- S-S-) can be reduced into the compound containing sulfydryl (SH).Cystamine (NH2CH2CH2SH)
Dimer (the NH of formation2CH2CH2S-S CH2CH2NH2) one end amino (NH2) with the carboxyl of graphene oxide it is combined into acid amides
Key (O=C-NH), the amino of the other end and the carboxyl of heparin are combined into amido link, and the compound that stops of formation is through anticancer in physical bond
After medicine, through blood arrival cancer cell, after intracellular of transduceing, in the presence of GSH, disulfide bond degraded, dissociate heparin, simultaneously
Because intracellular pH is relatively low, the medicine discharged increases, and heparin adds the therapeutic action of cancer therapy drug.
(3) graphene oxide after the present invention is heparin modified can be as the carrier material of medicine particularly cancer therapy drug.Profit
With pH sensitivity principles or redox principle, and adipic dihydrazide or cystamine are connection molecule, with heparin to graphite oxide
Alkene is chemically modified, and has the advantages of increase water solubility of graphene oxide, cancer therapy drug antitumor synergy, solves
Graphene oxide water solubility is poor, the problems such as easily assembling.It can specifically be combined, be pressed down with cytokine profiles using heparin simultaneously
Angiogenesis processed, play synergistic antitumor effect.
Brief description of the drawings
The structure of Fig. 1 graphene oxides, contain substantial amounts of carbon-carbon double bond in the structure, simultaneously containing hydroxyl (OH), carboxyl
And epoxy bond (C-O-C) (COOH).
The structure of Fig. 2 reproducibility graphene oxides, compared with graphene oxide, its oxy radical greatly reduces the structure,
There is substantial amounts of hydroxy-acid group at structural edge.
The structure of Fig. 3 heparin, the linear polysaccharide that the structure is made up of the glycuronic acid and gucosamine disaccharides of 1 → 4 connection
Class compound, each disaccharide unit contain a carboxyl, one or more hydroxyls, 2~2.5 sulfates, and each chain contains about
0.3 free amino.
The typical IR collection of illustrative plates of Fig. 4 graphene oxides, in the collection of illustrative plates, 3885cm-1Wider peak is attributed to hydroxyl (O-H)
Stretching vibration peak, due to containing water, hydrogen bond is formd with water, forms associative structure, therefore is wider peak；2975cm-1Peak,
2928cm-1Peak and 2892cm-1Peak is attributed to the stretching vibration peak of hydrocarbon (C-H), 1648cm-1Peak be attributed to carboxyl (O=C-
OH stretching vibration peak), 1420cm-1Peak and 1383cm-1Peak is attributed to carbon hydroxyl (C-OH) stretching vibration peak, 1090cm-1
Peak and 1050cm-1Peak is attributed to the stretching vibration peak of the carbon oxygen (C-O-C) of epoxide group.
The typical IR collection of illustrative plates of Fig. 5 reproducibility graphene oxides, in the collection of illustrative plates, 3885cm-1The wider peak in peak is attributed to hydroxyl
The stretching vibration peak of base (O-H), 2912cm-1Peak is attributed to the stretching vibration peak of hydrocarbon (C-H), 1631cm-1Peak is attributed to carboxylic
The stretching vibration peak of base (O=C-OH), 1574cm-1Peak is attributed to carbon-carbon double bond (C=C) stretching vibration peak of aromatic rings, carbon hydroxyl
The stretching vibration peak of base (C-OH), 1431cm-1Peak is attributed to carbon hydroxyl (C-OH) stretching vibration peak, 1060cm-1Peak and
1030cm-1Peak is attributed to the stretching vibration peak of the carbon oxygen (C-O-C) of epoxide group.
The typical IR collection of illustrative plates of the heparin of Fig. 6 adipic dihydrazides modification, in the collection of illustrative plates, 3423cm-1Peak and 3310cm-1
The two wider peaks of peak are attributed to the stretching vibration peak of hydroxyl (O-H) and amino (N-H), due to containing water, are formed with water
Hydrogen bond, forms associative structure, therefore be wider peak；2972cm-1Peak and 2934cm-1Peak is attributed to stretching for hydrocarbon (C-H)
Contracting vibration peak, 1655cm-1Peak is attributed to carboxyl (O=C-OH) asymmetric stretching vibration peak, 1701cm-1Peak is attributed to acid amides
I peaks, 1483cm-1Peak is attributed to acid amides II peaks, 1466cm-1Peak is attributed to the symmetrical stretching vibration peak of carboxyl (O=C-OH),
1228cm-1Peak is attributed to thiocarbonyl (S=O) asymmetric stretching vibration peak, 1032cm-1Peak and 1006cm-1Peak thiocarbonyl
(S=O) symmetrical stretching vibration peak.
The typical IR collection of illustrative plates of the LMWHs of Fig. 7 adipic dihydrazides modification, in the collection of illustrative plates, 3445cm-1Peak belongs to
For hydroxyl (O-H) and the stretching vibration peak of amino (N-H), due to containing water, hydrogen bond is formd with water, forms association knot
Structure, therefore be wider peak；2933cm-1Peak is attributed to the stretching vibration peak of hydrocarbon (C-H), 1660cm-1Peak is attributed to carboxyl (O
=C-OH) asymmetric stretching vibration peak, 1701cm-1Peak is attributed to amide I peaks, 1548cm-1Amino (N-H) on secondary amide
Stretching vibration and flexural vibrations (acid amides II peaks), 1385cm-1Peak is attributed to the symmetrical flexible of carboxyl (O=C-OH)
Vibration peak, 1228cm-1Peak is attributed to thiocarbonyl (S=O) asymmetric stretching vibration peak, 1031cm-1Peak and 985cm-1Peak sulphur
For the symmetrical stretching vibration peak of carbonyl (S=O).
The typical IR collection of illustrative plates of Fig. 8 heparin-adipic dihydrazide-reproducibility graphene oxides, in the collection of illustrative plates, 3441cm-1
Peak is attributed to the stretching vibration peak of hydroxyl (O-H) and amino (N-H)；2921cm-1Peak and 2848cm-1Peak is attributed to hydrocarbon
The stretching vibration peak of (C-H), 1641cm-1Peak is attributed to carboxyl (O=C-OH) asymmetric stretching vibration peak, 1549cm-1It is secondary
The stretching vibration and flexural vibrations (acid amides II peaks) of amino (N-H), 1385cm on acid amides-1Peak is attributed to carboxyl (O=
C-OH) symmetrical stretching vibration peak, 1212cm-1Peak is attributed to thiocarbonyl (S=O) asymmetric stretching vibration peak,
1026cm-1Peak and 1000cm-1The symmetrical stretching vibration peak of peak thiocarbonyl (S=O).
The typical IR collection of illustrative plates of Fig. 9 LMWHs-adipic dihydrazide-reproducibility graphene oxides, in the collection of illustrative plates,
3425cm-1Peak is attributed to the stretching vibration peak of hydroxyl (O-H) and amino (N-H)；2925cm-1Peak is attributed to hydrocarbon (C-H)
Stretching vibration peak, 1648cm-1Peak is attributed to carboxyl (O=C-OH) asymmetric stretching vibration peak, 1552cm-1Peak is attributed to
The stretching vibration and flexural vibrations (acid amides II peaks) of amino (N-H), 1447cm on secondary amide-1Peak is attributed to carboxyl (O
=C-OH) symmetrical stretching vibration peak, 1216cm-1Peak is attributed to thiocarbonyl (S=O) asymmetric stretching vibration peak,
1031cm-1Peak and 1000cm-1The symmetrical stretching vibration peak of peak thiocarbonyl (S=O).
Embodiment
A kind of heparin modified graphene oxide and preparation method thereof, this hair can be described in further detail by implementation below
It is bright, but the present invention is not limited in following embodiment.
Embodiment 1
Graphene oxide-heparin stops compound
(1) the preparation of graphene oxide (GO) takes graphite powder 0.40kg, adds 2.40 liters of the concentrated sulfuric acid, potassium peroxydisulfate slowly
0.60kg and phosphorus pentoxide 0.60kg, 80 DEG C are heated to, stir 10~24h, obtain navy blue mixed liquor, be gradually cooling to room
Temperature, 10 liters of purified water is added, filtering, collects residue, 60 DEG C are dried overnight, and obtain the graphite powder of pre-oxidation.Take the graphite of pre-oxidation
Powder 0.20kg, 15 liters (0 DEG C) are added in the concentrated sulfuric acid of ice bath slowly, under condition of ice bath, stir while the addition that blows slowly
Potassium permanganate 2.5kg.Continue to stir half an hour after adding, reaction solution gradually heats up be heated to 40 DEG C afterwards, stir 8h.Slowly
The mixed liquor for adding 3 liters of 75 liters of purified water and hydrogen peroxide (30wt%) terminates above-mentioned reaction.Stand overnight, question response liquid knot
Crystalline substance goes out precipitation, abandoning supernatant, respectively with dilute hydrochloric acid solution (1:10, v/v) and water washing precipitation is twice to three times, filtering, obtains
Precipitation, again with aqueous suspension, and dialysed with water, 2~3h of ultrasound (15 DEG C of solution temperature ＜), is obtained under obtained filtered fluid ice bath
The brown GO colloidal solution of normal distribution.
(2) the liquaemin 5g for preparing of heparin (ADH-UFH) of adipic dihydrazide modification is dissolved in 0.4 liter of water, adds oneself two
Acid dihydrazide (ADH) 2.2g, 1- ethyl-(3- dimethylaminopropyls) phosphinylidyne diimmonium salt hydrochlorate (EDCHCl) 3.2g and 1-
Hydroxybenzotriazole (HOBt) 2.2g, regulation solution ph to 6.8.Stirring reaction 24h at 25~30 DEG C.Unreacted reactant water is saturating
Analysis removes, freeze-drying, obtains product, and Fig. 6 is the typical IR collection of illustrative plates of the heparin of adipic dihydrazide modification.
(3) the GO (GO-ADH-UFH) of ADH-UFH modifications preparation takes 1 liter of 1mg/mL graphene oxide solutions, adds
EDCHCl 28.7g and n-hydroxysuccinimide sulfonate sodium (sulfur-NHS) 32.5g, adjust solution ph 6.4,25
Stirring reaction 2h at~30 DEG C, with activating carboxy acid's group.Add ADH-UFH 10g, stirring reaction 12h at 25~30 DEG C, dialysis
Unreacted starting material material is removed, GO-ADH-UFH is obtained and stops polymer solution.
Embodiment 2
Reproducibility graphene oxide-heparin stops compound
(1) the preparation of reproducibility graphene oxide (rGO) takes (1) middle 1 liter of the GO colloidal solution of above-described embodiment 1, is made
The 0.1g/L aqueous solution, adds vitamin C 2g, ultrasonic 30min at 60 DEG C, adds sodium chloride 5g afterwards, stands 2h, under 4000rpm
1000min is centrifuged, collects precipitation, is respectively washed 2 times with ethanol and pure water respectively, 1000min, reproducibility oxygen is centrifuged under 4000rpm
Graphite alkene, and with purifying aqueous suspension, ultrasonic 1h, obtained rGO colloidal solution.
(2) the rGO (rGO-ADH-UFH) of ADH-UFH modifications preparation takes 1 liter of 1mg/mL rGO solution, adds EDC
HCl 28.7g and sulfur-NHS 32.5g, stirring reaction 1.5h at 6.4,25~30 DEG C of solution ph is adjusted, with activating carboxy acid
Group.Add above-described embodiment 1 (2) middle ADH-UFH 10g, stirring reaction 20h at 25~30 DEG C, dialysis removes unreacted starting
Material, obtain rGO-ADH-UFH and stop polymer solution.Fig. 8 is the typical case of heparin-adipic dihydrazide-reproducibility graphene oxide
Infared spectrum.
Embodiment 3
Graphene oxide-LMWHs stops compound
(1) the preparation of the LMWHs (ADH-LMWH) of adipic dihydrazide modification takes low molecular sodium heparin 10g to be dissolved in
In 1 liter of water, ADH 2g, EDCHCl 8g and HOBt 6g, regulation solution ph to 6.6 are added.Stirring reaction at 25~30 DEG C
36h.Unreacted reactant is dialysed with water and removed, and freeze-drying, obtains product.Fig. 7 is the LMWHs of adipic dihydrazide modification
Typical IR collection of illustrative plates.
(2) the GO (GO-ADH-LMWH) of ADH-UFH modifications preparation takes the (1) middle 1mg/mL graphite oxides of above-described embodiment 1
1 liter of alkene solution, EDCHCl 26g and sulfur-NHS 30g are added, adjust stirring reaction at 6.2,25~30 DEG C of solution ph
2h, with activating carboxy acid's group.Add ADH-LMWH 10g, stirring reaction 18h at 25~30 DEG C, dialysis removes unreacted starting material
Material, obtain GO-ADH-LMWH and stop polymer solution.
Embodiment 4
Graphene oxide-LMWHs stops compound
(1) the preparation of the Enoxaparin Sodium (ADH-Eno) of adipic dihydrazide modification takes Enoxaparin Sodium (Eno) 12g molten
In 1 liter of water, ADH 4g, EDCHCl 3g and HOBt 3g, regulation solution ph to 6.6 are added.Stirred at 25~30 DEG C anti-
Answer 40h.Unreacted reactant is dialysed with water and removed, and freeze-drying, obtains product.
(2) the GO (GO-ADH-Eno) of ADH-UFH modifications preparation takes the (1) middle 1mg/mL graphene oxides of above-described embodiment 1
1.2 liters of solution, EDCHCl 26g and sulfur-NHS 30g are added, adjust stirring reaction at 6.2,25~30 DEG C of solution ph
2h, with activating carboxy acid's group.Add ADH-Eno 12g, stirring reaction 18h at 25~30 DEG C, dialysis removes unreacted starting material
Material, obtain GO-ADH-Eno and stop polymer solution.
Embodiment 5
Reproducibility graphene oxide-LMWHs stops compound
(1) the low molecular sodium heparin 10g for preparing of the LMWHs (ADH-LMWH) of adipic dihydrazide modification is dissolved in water 1
In liter, ADH 3g, EDCHCl 3g and HOBt 3g, regulation solution ph to 6.2 are added.Stirring reaction 36h at 25~30 DEG C.
Unreacted reactant is dialysed with water and removed, and freeze-drying, obtains product.
(2) the rGO (rGO-ADH-LMWH) of ADH-LMWH modifications preparation takes above-described embodiment 2 (1) middle 1mg/mL rGO is molten
1 liter of liquid, EDCHCl 30g and sulfur-NHS 35g are added, adjust stirring reaction 2h at 6.5,25~30 DEG C of solution ph.
ADH-LMWH 10g in addition, stirring reaction 12h at 25~30 DEG C, dialysis remove unreacted starting material material, obtain rGO-ADH-
UFH stops polymer solution.Fig. 9 is the typical IR collection of illustrative plates of LMWHs-adipic dihydrazide-reproducibility graphene oxide.
Embodiment 6
Graphene oxide-heparin stops compound
(1) the liquaemin 3g for preparing of the heparin (Cys-UFH) of cystamine modification is dissolved in 0.2 liter of water, adds cystamine 2g, EDC
HCl 2g and HOBt 2g, regulation solution ph to 6.5.Stirring reaction 24h at 25~30 DEG C.Unreacted reactant is dialysed with water and removed,
Freeze-drying, obtains product.
(2) the GO (GO-ss-UFH) of Cys-UFH modifications preparation takes 1 liter of 1mg/mL graphene oxide solutions, adds
EDCHCl 28.7g and sulfur-NHS 32.5g, stirring reaction 2h at 6.4,25~30 DEG C of solution ph is adjusted, with activation
Hydroxy-acid group.Add Cys-UFH 10g, stirring reaction 23h at 25~30 DEG C, dialysis removes unreacted starting material material, obtains GO-
Ss-UFH stops polymer solution.
Embodiment 7
Reproducibility graphene oxide-heparin stops compound
(1) the liquaemin 3g for preparing of the heparin (Cys-UFH) of cystamine modification is dissolved in 0.3 liter of water, adds cystamine 2.5g,
EDCHCl 3g and HOBt 3g, regulation solution ph to 6.5.Stirring reaction 30h at 25~30 DEG C.Unreacted reactant is dialysed with water
Remove, freeze-drying, obtain product.
(2) the rGO (rGO-ss-UFH) of Cys-UFH modifications preparation takes the (1) middle 1mg/mL rGO solution 1 of above-described embodiment 2
Rise, add EDCHCl 30g and sulfur-NHS 40g, stirring reaction 2h at 6.4,25~30 DEG C of solution ph is adjusted, with work
Change hydroxy-acid group.Add Cys-UFH 10g, stirring reaction 22h at 25~30 DEG C, dialysis removes unreacted starting material material, obtained
RGO-ss-UFH stops polymer solution.
Embodiment 8
Graphene oxide-LMWHs stops compound
(1) the preparation of the LMWHs (Cys-LMWH) of cystamine modification takes low molecular sodium heparin 3g to be dissolved in 0.6 liter of water,
Add cystamine 1.5g, EDCHCl 1.5g and HOBt 1.5g, regulation solution ph to 6.5.Stirring reaction 40h at 25~30 DEG C.
Unreacted reactant is dialysed with water and removed, and freeze-drying, obtains product.
(2) the GO (GO-ss-LMWH) of Cys-LMWH modifications preparation takes the (1) middle 1mg/mL graphite oxides of above-described embodiment 1
1 liter of alkene solution, EDCHCl 26g and sulfur-NHS 30g are added, adjust stirring reaction at 6.2,25~30 DEG C of solution ph
2h, with activating carboxy acid's group.Add Cys-LMWH 7g, stirring reaction 20h at 25~30 DEG C, dialysis removes unreacted starting material
Material, obtain GO-ss-LMWH and stop polymer solution.
Embodiment 9
Graphene oxide-LMWHs stops compound
(1) the preparation of the Parnaparin Sodium (Cys-Par) of cystamine modification takes Parnaparin Sodium (Par) 3g to be dissolved in 0.2 liter of water, adds
Cystamine 2g, EDCHCl 2g and HOBt 2g, regulation solution ph to 6.5.Stirring reaction 45h at 25~30 DEG C.Unreacted reactant
Dialysed and removed with water, freeze-drying, obtain product.
(2) the GO (GO-ss-Par) of Cys-Par modifications preparation takes the (1) middle 1mg/mL graphene oxides of above-described embodiment 1
1 liter of solution, EDCHCl 26g and sulfur-NHS 30g are added, adjust stirring reaction at 6.2,25~30 DEG C of solution ph
2h, with activating carboxy acid's group.Add Cys-Par 8g, stirring reaction 22h at 25~30 DEG C, dialysis removes unreacted starting material
Material, obtain GO-ss-Par and stop polymer solution.
Embodiment 10
Reproducibility graphene oxide-LMWHs stops compound
(1) the preparation of the LMWHs (Cys-LMWH) of cystamine modification takes low molecular sodium heparin 3g to be dissolved in 0.25 liter of water
In, add cystamine 2.5g, EDCHCl 1g and HOBt 1g, regulation solution ph to 6.6.Stirring reaction 36h at 25~30 DEG C.Not
Reactant is dialysed with water and removed, and freeze-drying, obtains product.
(2) the rGO (rGO-ss-LMWH) of Cys-LMWH modifications preparation takes above-described embodiment 2 (1) middle 1mg/mL rGO is molten
1 liter of liquid, EDCHCl 30g and sulfur-NHS 35g are added, adjust stirring reaction 2h at 6.5,25~30 DEG C of solution ph.
Cys-LMWH 10g in addition, stirring reaction 22h at 25~30 DEG C, dialysis remove unreacted starting material material, obtain rGO-ss-
UFH stops polymer solution.
Although above-mentioned the embodiment of the present invention is described with reference to accompanying drawing, model not is protected to the present invention
The limitation enclosed, one of ordinary skill in the art should be understood that on the basis of technical scheme those skilled in the art are not
Need to pay various modifications or deformation that creative work can make still within protection scope of the present invention.
Claims (7)
1. a kind of heparin or the graphene oxide of its salt modification, it is characterized in that：It is to be prepared by the following method to obtain：
Using adipic dihydrazide as connection molecule, between heparin and graphene oxide by amido link be connected obtain it is heparin modified
Graphene oxide；
Comprise the concrete steps that：Heparin is modified using adipic dihydrazide, the heparin after adipic dihydrazide modification is obtained, then incites somebody to action
Heparin after the adipic dihydrazide modification arrived and graphene oxide after activation react 6 ~ 24h at 25 ~ 30 DEG C, obtain with oneself two
Acid dihydrazide stops polymer solution as the heparin modified graphene oxide of connection molecule.
2. heparin as claimed in claim 1 or the graphene oxide of its salt modification, it is characterized in that：The adipic dihydrazide is repaiied
Decorations heparin method be：Liquaemin is soluble in water, adds adipic dihydrazide, 1- ethyls-(3- dimethylaminopropyls) phosphinylidyne
Diimmonium salt hydrochlorate and I-hydroxybenzotriazole, regulation solution ph to 6.0~7.0；At 25~30 DEG C stirring reaction 12~
48h；Unreacted starting material dialysis removes, and dries, and obtains the heparin after the modification of product adipic dihydrazide.
3. heparin as claimed in claim 1 or the graphene oxide of its salt modification, it is characterized in that：Take graphene oxide solution,
Add 1- ethyls-(3- dimethylaminopropyls) phosphinylidyne diimmonium salt hydrochlorate and n-hydroxysuccinimide sulfonate sodium, regulation
Solution ph is to 6.0~7.0, stirring reaction 1~3 hour at 25~30 DEG C, the graphene oxide after being activated.
4. heparin as claimed in claim 2 or the graphene oxide of its salt modification, it is characterized in that：The liquaemin refers to general
Logical liquaemin, low molecular sodium heparin one or two kinds of combination therein；The medicinal forms of the low molecular sodium heparin are medicinal
Low molecular sodium heparin, Certoparin Sodium, Bemiparin sodium, Dalteparin Sodium, Enoxaparin Sodium, Parnaparin Sodium, Clivarin sodium, booth
Prick the one or more kinds of combination of liquaemin, Ardeparin Sodium.
5. the graphene oxide of the heparin or the modification of its salt as any one of claim 1 or 2, it is characterized in that：The oxygen
Graphite alkene is reproducibility graphene oxide of the graphene oxide after reducing agent reduces.
6. the graphene oxide of the heparin or the modification of its salt any one of claim 1 ~ 5 is preparing answering for pharmaceutical carrier
With.
7. one kind contains anticancer drug complex, it is characterized in that, it is prepared by the following method to obtain：Appoint in claim 1 ~ 5
The graphene oxide of heparin or the modification of its salt described in one mixes with cancer therapy drug, stirs, dialysis, removes the medicine of unentrapped
Thing, obtain anticancer drug complex.
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| CN105688226B (en) * | 2016-03-09 | 2017-05-10 | 西北大学 | Intelligent GO (graphene oxide) drug carrier with supermolecule sensitive detaching capacity, preparation method and application |
| WO2018032502A1 (en) * | 2016-08-19 | 2018-02-22 | 苏州融析生物科技有限公司 | Sheep-derived low molecular weight heparin, preparation method therefor and application thereof |
| CN107759712B (en) * | 2016-08-19 | 2020-03-24 | 苏州融析生物科技有限公司 | Sheep-derived low-molecular-weight heparin and preparation method and application thereof |
| CN107137788B (en) * | 2017-04-28 | 2019-05-24 | 淮阴工学院 | A method of chitosan/test tube of hepari graphene oxide composite multilayer membrane is prepared in medical magnesium alloy surface |
| CN114479390B (en) * | 2022-01-26 | 2023-06-23 | 安徽理工大学 | Preparation method of graphene oxide/phenol red type polyarylester composite material |
| CN114479391B (en) * | 2022-01-26 | 2023-06-23 | 安徽理工大学 | Preparation method of graphene oxide/bisphenol-A type polyarylate composite material |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| KR20110031826A (en) * | 2009-09-21 | 2011-03-29 | 성균관대학교산학협력단 | Graphene/biopolymer nanofiber composites and preparation method thereof |
| CN103100114A (en) * | 2013-01-23 | 2013-05-15 | 西南交通大学 | Preparation method of medical metal surface slow-released growth factor coating |
| CN104826128A (en) * | 2015-04-30 | 2015-08-12 | 中国药科大学 | Polysaccharide modified reduction-sensitive graphene oxide carrier with organism lesion site triggered drug release and preparation and application of pharmaceutical composition thereof |
-
2015
- 2015-11-18 CN CN201510796642.1A patent/CN105343890B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20110031826A (en) * | 2009-09-21 | 2011-03-29 | 성균관대학교산학협력단 | Graphene/biopolymer nanofiber composites and preparation method thereof |
| CN103100114A (en) * | 2013-01-23 | 2013-05-15 | 西南交通大学 | Preparation method of medical metal surface slow-released growth factor coating |
| CN104826128A (en) * | 2015-04-30 | 2015-08-12 | 中国药科大学 | Polysaccharide modified reduction-sensitive graphene oxide carrier with organism lesion site triggered drug release and preparation and application of pharmaceutical composition thereof |
Non-Patent Citations (2)
| Title |
|---|
| Blood Compatible Graphene/Heparin Conjugate through Noncovalent Chemistry;Da Young Lee等;《Biomacromolecules》;20110110;第12卷;第336-341页 * |
| 基于氧化石墨烯的类肝素水凝胶及其血液相容性研究;何超等;《2015年全国高分子学术论文报告会》;20151021;第BP-073页 * |
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