WO2015039555A1 - 基于氧化石墨烯材料的造影剂及其制备方法 - Google Patents
基于氧化石墨烯材料的造影剂及其制备方法 Download PDFInfo
- Publication number
- WO2015039555A1 WO2015039555A1 PCT/CN2014/085611 CN2014085611W WO2015039555A1 WO 2015039555 A1 WO2015039555 A1 WO 2015039555A1 CN 2014085611 W CN2014085611 W CN 2014085611W WO 2015039555 A1 WO2015039555 A1 WO 2015039555A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- contrast agent
- molecule
- graphene oxide
- group
- bridging
- Prior art date
Links
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 title claims abstract description 92
- 239000002872 contrast media Substances 0.000 title claims abstract description 92
- 229910021389 graphene Inorganic materials 0.000 title claims abstract description 78
- 238000002360 preparation method Methods 0.000 title claims abstract description 19
- 239000000463 material Substances 0.000 title abstract description 6
- -1 gadolinium metal complex Chemical group 0.000 claims abstract description 28
- 239000000126 substance Substances 0.000 claims abstract description 22
- 229910052688 Gadolinium Inorganic materials 0.000 claims abstract 7
- 230000000536 complexating effect Effects 0.000 claims description 22
- AOJJSUZBOXZQNB-TZSSRYMLSA-N Doxorubicin Chemical compound O([C@H]1C[C@@](O)(CC=2C(O)=C3C(=O)C=4C=CC=C(C=4C(=O)C3=C(O)C=21)OC)C(=O)CO)[C@H]1C[C@H](N)[C@H](O)[C@H](C)O1 AOJJSUZBOXZQNB-TZSSRYMLSA-N 0.000 claims description 18
- 238000000034 method Methods 0.000 claims description 15
- 125000000524 functional group Chemical group 0.000 claims description 14
- 238000003384 imaging method Methods 0.000 claims description 14
- 230000008685 targeting Effects 0.000 claims description 14
- 239000000203 mixture Substances 0.000 claims description 13
- 239000002105 nanoparticle Substances 0.000 claims description 12
- QPCDCPDFJACHGM-UHFFFAOYSA-N N,N-bis{2-[bis(carboxymethyl)amino]ethyl}glycine Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(=O)O)CCN(CC(O)=O)CC(O)=O QPCDCPDFJACHGM-UHFFFAOYSA-N 0.000 claims description 10
- 229960004679 doxorubicin Drugs 0.000 claims description 9
- 229960003330 pentetic acid Drugs 0.000 claims description 9
- 229920001223 polyethylene glycol Polymers 0.000 claims description 9
- 239000002202 Polyethylene glycol Substances 0.000 claims description 8
- 239000000975 dye Substances 0.000 claims description 8
- 239000000700 radioactive tracer Substances 0.000 claims description 8
- 229940126585 therapeutic drug Drugs 0.000 claims description 8
- 239000003937 drug carrier Substances 0.000 claims description 7
- 239000008194 pharmaceutical composition Substances 0.000 claims description 7
- KIUKXJAPPMFGSW-DNGZLQJQSA-N (2S,3S,4S,5R,6R)-6-[(2S,3R,4R,5S,6R)-3-Acetamido-2-[(2S,3S,4R,5R,6R)-6-[(2R,3R,4R,5S,6R)-3-acetamido-2,5-dihydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-2-carboxy-4,5-dihydroxyoxan-3-yl]oxy-5-hydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-3,4,5-trihydroxyoxane-2-carboxylic acid Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@H](O3)C(O)=O)O)[C@H](O)[C@@H](CO)O2)NC(C)=O)[C@@H](C(O)=O)O1 KIUKXJAPPMFGSW-DNGZLQJQSA-N 0.000 claims description 5
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 5
- KLWPJMFMVPTNCC-UHFFFAOYSA-N Camptothecin Natural products CCC1(O)C(=O)OCC2=C1C=C3C4Nc5ccccc5C=C4CN3C2=O KLWPJMFMVPTNCC-UHFFFAOYSA-N 0.000 claims description 5
- 229920002307 Dextran Polymers 0.000 claims description 5
- 239000005411 L01XE02 - Gefitinib Substances 0.000 claims description 5
- 229920002873 Polyethylenimine Polymers 0.000 claims description 5
- 239000004480 active ingredient Substances 0.000 claims description 5
- VSJKWCGYPAHWDS-FQEVSTJZSA-N camptothecin Chemical compound C1=CC=C2C=C(CN3C4=CC5=C(C3=O)COC(=O)[C@]5(O)CC)C4=NC2=C1 VSJKWCGYPAHWDS-FQEVSTJZSA-N 0.000 claims description 5
- 229940127093 camptothecin Drugs 0.000 claims description 5
- 239000000306 component Substances 0.000 claims description 5
- 239000000412 dendrimer Substances 0.000 claims description 5
- 229920000736 dendritic polymer Polymers 0.000 claims description 5
- VSJKWCGYPAHWDS-UHFFFAOYSA-N dl-camptothecin Natural products C1=CC=C2C=C(CN3C4=CC5=C(C3=O)COC(=O)C5(O)CC)C4=NC2=C1 VSJKWCGYPAHWDS-UHFFFAOYSA-N 0.000 claims description 5
- SZVJSHCCFOBDDC-UHFFFAOYSA-N ferrosoferric oxide Chemical compound O=[Fe]O[Fe]O[Fe]=O SZVJSHCCFOBDDC-UHFFFAOYSA-N 0.000 claims description 5
- XGALLCVXEZPNRQ-UHFFFAOYSA-N gefitinib Chemical compound C=12C=C(OCCCN3CCOCC3)C(OC)=CC2=NC=NC=1NC1=CC=C(F)C(Cl)=C1 XGALLCVXEZPNRQ-UHFFFAOYSA-N 0.000 claims description 5
- 229920002674 hyaluronan Polymers 0.000 claims description 5
- 229960003160 hyaluronic acid Drugs 0.000 claims description 5
- 229940084651 iressa Drugs 0.000 claims description 5
- 239000012099 Alexa Fluor family Substances 0.000 claims description 4
- 229920001661 Chitosan Polymers 0.000 claims description 4
- PWHULOQIROXLJO-UHFFFAOYSA-N Manganese Chemical compound [Mn] PWHULOQIROXLJO-UHFFFAOYSA-N 0.000 claims description 4
- WDLRUFUQRNWCPK-UHFFFAOYSA-N Tetraxetan Chemical compound OC(=O)CN1CCN(CC(O)=O)CCN(CC(O)=O)CCN(CC(O)=O)CC1 WDLRUFUQRNWCPK-UHFFFAOYSA-N 0.000 claims description 4
- 238000006243 chemical reaction Methods 0.000 claims description 4
- MHMNJMPURVTYEJ-UHFFFAOYSA-N fluorescein-5-isothiocyanate Chemical compound O1C(=O)C2=CC(N=C=S)=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 MHMNJMPURVTYEJ-UHFFFAOYSA-N 0.000 claims description 4
- 150000004676 glycans Chemical class 0.000 claims description 4
- 229910052748 manganese Inorganic materials 0.000 claims description 4
- 239000011572 manganese Substances 0.000 claims description 4
- 229910001437 manganese ion Inorganic materials 0.000 claims description 4
- 102000039446 nucleic acids Human genes 0.000 claims description 4
- 108020004707 nucleic acids Proteins 0.000 claims description 4
- 150000007523 nucleic acids Chemical class 0.000 claims description 4
- 229920001282 polysaccharide Polymers 0.000 claims description 4
- 239000005017 polysaccharide Substances 0.000 claims description 4
- 102000004196 processed proteins & peptides Human genes 0.000 claims description 4
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 4
- PYWVYCXTNDRMGF-UHFFFAOYSA-N rhodamine B Chemical compound [Cl-].C=12C=CC(=[N+](CC)CC)C=C2OC2=CC(N(CC)CC)=CC=C2C=1C1=CC=CC=C1C(O)=O PYWVYCXTNDRMGF-UHFFFAOYSA-N 0.000 claims description 4
- 239000012266 salt solution Substances 0.000 claims description 4
- 230000001588 bifunctional effect Effects 0.000 claims description 3
- 229920001184 polypeptide Polymers 0.000 claims description 3
- 108090000623 proteins and genes Proteins 0.000 claims description 3
- 102000004169 proteins and genes Human genes 0.000 claims description 3
- 239000003085 diluting agent Substances 0.000 claims description 2
- 239000005426 pharmaceutical component Substances 0.000 claims description 2
- 150000000921 Gadolinium Chemical class 0.000 claims 4
- UIWYJDYFSGRHKR-UHFFFAOYSA-N gadolinium atom Chemical compound [Gd] UIWYJDYFSGRHKR-UHFFFAOYSA-N 0.000 claims 3
- INVGWHRKADIJHF-UHFFFAOYSA-N Sanguinarin Chemical compound C1=C2OCOC2=CC2=C3[N+](C)=CC4=C(OCO5)C5=CC=C4C3=CC=C21 INVGWHRKADIJHF-UHFFFAOYSA-N 0.000 claims 2
- 229910052751 metal Inorganic materials 0.000 claims 2
- 239000002184 metal Substances 0.000 claims 2
- UGFAIRIUMAVXCW-UHFFFAOYSA-N Carbon monoxide Chemical compound [O+]#[C-] UGFAIRIUMAVXCW-UHFFFAOYSA-N 0.000 claims 1
- FCEXWTOTHXCQCQ-UHFFFAOYSA-N Ethoxydihydrosanguinarine Natural products C12=CC=C3OCOC3=C2C(OCC)N(C)C(C2=C3)=C1C=CC2=CC1=C3OCO1 FCEXWTOTHXCQCQ-UHFFFAOYSA-N 0.000 claims 1
- MEANOSLIBWSCIT-UHFFFAOYSA-K gadolinium trichloride Chemical compound Cl[Gd](Cl)Cl MEANOSLIBWSCIT-UHFFFAOYSA-K 0.000 claims 1
- MWFSXYMZCVAQCC-UHFFFAOYSA-N gadolinium(iii) nitrate Chemical compound [Gd+3].[O-][N+]([O-])=O.[O-][N+]([O-])=O.[O-][N+]([O-])=O MWFSXYMZCVAQCC-UHFFFAOYSA-N 0.000 claims 1
- 229920002521 macromolecule Polymers 0.000 claims 1
- 229940084560 sanguinarine Drugs 0.000 claims 1
- YZRQUTZNTDAYPJ-UHFFFAOYSA-N sanguinarine pseudobase Natural products C1=C2OCOC2=CC2=C3N(C)C(O)C4=C(OCO5)C5=CC=C4C3=CC=C21 YZRQUTZNTDAYPJ-UHFFFAOYSA-N 0.000 claims 1
- 239000000243 solution Substances 0.000 description 16
- 239000002405 nuclear magnetic resonance imaging agent Substances 0.000 description 14
- KJTLSVCANCCWHF-UHFFFAOYSA-N Ruthenium Chemical group [Ru] KJTLSVCANCCWHF-UHFFFAOYSA-N 0.000 description 13
- 229940079593 drug Drugs 0.000 description 12
- 239000003814 drug Substances 0.000 description 12
- 238000003756 stirring Methods 0.000 description 9
- LMDZBCPBFSXMTL-UHFFFAOYSA-N 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide Chemical compound CCN=C=NCCCN(C)C LMDZBCPBFSXMTL-UHFFFAOYSA-N 0.000 description 8
- 230000000694 effects Effects 0.000 description 8
- 238000000502 dialysis Methods 0.000 description 7
- 229910052684 Cerium Inorganic materials 0.000 description 6
- OVBPIULPVIDEAO-LBPRGKRZSA-N folic acid Chemical compound C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-LBPRGKRZSA-N 0.000 description 6
- 238000011068 loading method Methods 0.000 description 5
- 238000001179 sorption measurement Methods 0.000 description 5
- 230000003834 intracellular effect Effects 0.000 description 4
- 238000012986 modification Methods 0.000 description 4
- 230000004048 modification Effects 0.000 description 4
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 description 4
- 150000004714 phosphonium salts Chemical class 0.000 description 4
- 229910052707 ruthenium Inorganic materials 0.000 description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 4
- UHOVQNZJYSORNB-UHFFFAOYSA-N Benzene Chemical compound C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 description 3
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 3
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 3
- OVBPIULPVIDEAO-UHFFFAOYSA-N N-Pteroyl-L-glutaminsaeure Natural products C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)NC(CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-UHFFFAOYSA-N 0.000 description 3
- 206010028980 Neoplasm Diseases 0.000 description 3
- 239000004698 Polyethylene Substances 0.000 description 3
- 125000000217 alkyl group Chemical group 0.000 description 3
- 125000004432 carbon atom Chemical group C* 0.000 description 3
- 238000012377 drug delivery Methods 0.000 description 3
- 229960000304 folic acid Drugs 0.000 description 3
- 235000019152 folic acid Nutrition 0.000 description 3
- 239000011724 folic acid Substances 0.000 description 3
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 3
- 229920000573 polyethylene Polymers 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 125000003837 (C1-C20) alkyl group Chemical group 0.000 description 2
- LDGWQMRUWMSZIU-LQDDAWAPSA-M 2,3-bis[(z)-octadec-9-enoxy]propyl-trimethylazanium;chloride Chemical compound [Cl-].CCCCCCCC\C=C/CCCCCCCCOCC(C[N+](C)(C)C)OCCCCCCCC\C=C/CCCCCCCC LDGWQMRUWMSZIU-LQDDAWAPSA-M 0.000 description 2
- HHLZCENAOIROSL-UHFFFAOYSA-N 2-[4,7-bis(carboxymethyl)-1,4,7,10-tetrazacyclododec-1-yl]acetic acid Chemical compound OC(=O)CN1CCNCCN(CC(O)=O)CCN(CC(O)=O)CC1 HHLZCENAOIROSL-UHFFFAOYSA-N 0.000 description 2
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 2
- WZUVPPKBWHMQCE-UHFFFAOYSA-N Haematoxylin Chemical compound C12=CC(O)=C(O)C=C2CC2(O)C1C1=CC=C(O)C(O)=C1OC2 WZUVPPKBWHMQCE-UHFFFAOYSA-N 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 2
- 229930012538 Paclitaxel Natural products 0.000 description 2
- 235000016551 Potentilla erecta Nutrition 0.000 description 2
- 241000372442 Wachendorfia thyrsiflora Species 0.000 description 2
- 239000007864 aqueous solution Substances 0.000 description 2
- 239000010953 base metal Substances 0.000 description 2
- 201000011510 cancer Diseases 0.000 description 2
- 238000004113 cell culture Methods 0.000 description 2
- GWXLDORMOJMVQZ-UHFFFAOYSA-N cerium Chemical compound [Ce] GWXLDORMOJMVQZ-UHFFFAOYSA-N 0.000 description 2
- VYLVYHXQOHJDJL-UHFFFAOYSA-K cerium trichloride Chemical compound Cl[Ce](Cl)Cl VYLVYHXQOHJDJL-UHFFFAOYSA-K 0.000 description 2
- HSJPMRKMPBAUAU-UHFFFAOYSA-N cerium(3+);trinitrate Chemical compound [Ce+3].[O-][N+]([O-])=O.[O-][N+]([O-])=O.[O-][N+]([O-])=O HSJPMRKMPBAUAU-UHFFFAOYSA-N 0.000 description 2
- 230000015271 coagulation Effects 0.000 description 2
- 238000005345 coagulation Methods 0.000 description 2
- 150000004696 coordination complex Chemical class 0.000 description 2
- 150000002148 esters Chemical class 0.000 description 2
- 239000012091 fetal bovine serum Substances 0.000 description 2
- 229910002804 graphite Inorganic materials 0.000 description 2
- 239000010439 graphite Substances 0.000 description 2
- 229910052736 halogen Inorganic materials 0.000 description 2
- 229960003350 isoniazid Drugs 0.000 description 2
- QRXWMOHMRWLFEY-UHFFFAOYSA-N isoniazide Chemical compound NNC(=O)C1=CC=NC=C1 QRXWMOHMRWLFEY-UHFFFAOYSA-N 0.000 description 2
- 239000003446 ligand Substances 0.000 description 2
- 238000002595 magnetic resonance imaging Methods 0.000 description 2
- 230000014759 maintenance of location Effects 0.000 description 2
- 230000004060 metabolic process Effects 0.000 description 2
- 239000012074 organic phase Substances 0.000 description 2
- 229960001592 paclitaxel Drugs 0.000 description 2
- 239000002245 particle Substances 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 229910052703 rhodium Inorganic materials 0.000 description 2
- 239000010948 rhodium Substances 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- 125000001424 substituent group Chemical group 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N sulfuric acid Substances OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- RCINICONZNJXQF-MZXODVADSA-N taxol Chemical compound O([C@@H]1[C@@]2(C[C@@H](C(C)=C(C2(C)C)[C@H](C([C@]2(C)[C@@H](O)C[C@H]3OC[C@]3([C@H]21)OC(C)=O)=O)OC(=O)C)OC(=O)[C@H](O)[C@@H](NC(=O)C=1C=CC=CC=1)C=1C=CC=CC=1)O)C(=O)C1=CC=CC=C1 RCINICONZNJXQF-MZXODVADSA-N 0.000 description 2
- 239000003981 vehicle Substances 0.000 description 2
- QBPPRVHXOZRESW-UHFFFAOYSA-N 1,4,7,10-tetraazacyclododecane Chemical compound C1CNCCNCCNCCN1 QBPPRVHXOZRESW-UHFFFAOYSA-N 0.000 description 1
- HSXRCEVTMOWCFI-UHFFFAOYSA-N 2-[4,7,10-tris(carboxymethyl)-6-methyl-1,4,7,10-tetrazacyclododec-1-yl]acetic acid Chemical compound CC1CN(CC(O)=O)CCN(CC(O)=O)CCN(CC(O)=O)CCN1CC(O)=O HSXRCEVTMOWCFI-UHFFFAOYSA-N 0.000 description 1
- UOQHWNPVNXSDDO-UHFFFAOYSA-N 3-bromoimidazo[1,2-a]pyridine-6-carbonitrile Chemical compound C1=CC(C#N)=CN2C(Br)=CN=C21 UOQHWNPVNXSDDO-UHFFFAOYSA-N 0.000 description 1
- 229920000936 Agarose Polymers 0.000 description 1
- 150000000703 Cerium Chemical class 0.000 description 1
- RPNUMPOLZDHAAY-UHFFFAOYSA-N Diethylenetriamine Chemical compound NCCNCCN RPNUMPOLZDHAAY-UHFFFAOYSA-N 0.000 description 1
- RWSOTUBLDIXVET-UHFFFAOYSA-N Dihydrogen sulfide Chemical class S RWSOTUBLDIXVET-UHFFFAOYSA-N 0.000 description 1
- AEMRFAOFKBGASW-UHFFFAOYSA-N Glycolic acid Natural products OCC(O)=O AEMRFAOFKBGASW-UHFFFAOYSA-N 0.000 description 1
- IQUHNCOJRJBMSU-UHFFFAOYSA-N H3HP-DO3A Chemical compound CC(O)CN1CCN(CC(O)=O)CCN(CC(O)=O)CCN(CC(O)=O)CC1 IQUHNCOJRJBMSU-UHFFFAOYSA-N 0.000 description 1
- PEEHTFAAVSWFBL-UHFFFAOYSA-N Maleimide Chemical compound O=C1NC(=O)C=C1 PEEHTFAAVSWFBL-UHFFFAOYSA-N 0.000 description 1
- 229920003171 Poly (ethylene oxide) Polymers 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
- BLRPTPMANUNPDV-UHFFFAOYSA-N Silane Chemical compound [SiH4] BLRPTPMANUNPDV-UHFFFAOYSA-N 0.000 description 1
- MRMOMIYVTPUCMS-UHFFFAOYSA-N acetic acid;pyrrolidine-2,5-dione Chemical compound CC(O)=O.O=C1CCC(=O)N1 MRMOMIYVTPUCMS-UHFFFAOYSA-N 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 150000008064 anhydrides Chemical class 0.000 description 1
- 239000008346 aqueous phase Substances 0.000 description 1
- 150000001540 azides Chemical class 0.000 description 1
- 238000004166 bioassay Methods 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- PMSGJXMYHUSZEI-UHFFFAOYSA-N butanedioic acid;pyrrolidine-2,5-dione Chemical compound O=C1CCC(=O)N1.OC(=O)CCC(O)=O PMSGJXMYHUSZEI-UHFFFAOYSA-N 0.000 description 1
- 125000000484 butyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 1
- 239000012930 cell culture fluid Substances 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 125000000753 cycloalkyl group Chemical group 0.000 description 1
- 125000001559 cyclopropyl group Chemical group [H]C1([H])C([H])([H])C1([H])* 0.000 description 1
- 125000002704 decyl group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])* 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- YWEUIGNSBFLMFL-UHFFFAOYSA-N diphosphonate Chemical compound O=P(=O)OP(=O)=O YWEUIGNSBFLMFL-UHFFFAOYSA-N 0.000 description 1
- 239000006185 dispersion Substances 0.000 description 1
- 238000003255 drug test Methods 0.000 description 1
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000012894 fetal calf serum Substances 0.000 description 1
- 125000003929 folic acid group Chemical group 0.000 description 1
- 125000002485 formyl group Chemical class [H]C(*)=O 0.000 description 1
- 125000005843 halogen group Chemical group 0.000 description 1
- 150000002367 halogens Chemical class 0.000 description 1
- 125000004435 hydrogen atom Chemical group [H]* 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000005865 ionizing radiation Effects 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 229910052741 iridium Inorganic materials 0.000 description 1
- 125000000959 isobutyl group Chemical group [H]C([H])([H])C([H])(C([H])([H])[H])C([H])([H])* 0.000 description 1
- 239000012948 isocyanate Substances 0.000 description 1
- 150000002513 isocyanates Chemical class 0.000 description 1
- 125000001449 isopropyl group Chemical group [H]C([H])([H])C([H])(*)C([H])([H])[H] 0.000 description 1
- 229940099563 lactobionic acid Drugs 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 229910021645 metal ion Inorganic materials 0.000 description 1
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 1
- VNXBKJFUJUWOCW-UHFFFAOYSA-N methylcyclopropane Chemical group CC1CC1 VNXBKJFUJUWOCW-UHFFFAOYSA-N 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 150000002772 monosaccharides Chemical class 0.000 description 1
- 239000002086 nanomaterial Substances 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 239000011368 organic material Substances 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- QZPDFSCRLSFIBE-UHFFFAOYSA-N pentanoic acid;pyrrolidine-2,5-dione Chemical compound CCCCC(O)=O.O=C1CCC(=O)N1 QZPDFSCRLSFIBE-UHFFFAOYSA-N 0.000 description 1
- DLYUQMMRRRQYAE-UHFFFAOYSA-N phosphorus pentoxide Inorganic materials O1P(O2)(=O)OP3(=O)OP1(=O)OP2(=O)O3 DLYUQMMRRRQYAE-UHFFFAOYSA-N 0.000 description 1
- 229920000747 poly(lactic acid) Polymers 0.000 description 1
- 239000004626 polylactic acid Substances 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- USHAGKDGDHPEEY-UHFFFAOYSA-L potassium persulfate Chemical compound [K+].[K+].[O-]S(=O)(=O)OOS([O-])(=O)=O USHAGKDGDHPEEY-UHFFFAOYSA-L 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- XANRGTLFBAJYDA-UHFFFAOYSA-N propanoic acid;pyrrolidine-2,5-dione Chemical compound CCC(O)=O.O=C1CCC(=O)N1 XANRGTLFBAJYDA-UHFFFAOYSA-N 0.000 description 1
- 125000001436 propyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- GTCKPGDAPXUISX-UHFFFAOYSA-N ruthenium(3+);trinitrate Chemical compound [Ru+3].[O-][N+]([O-])=O.[O-][N+]([O-])=O.[O-][N+]([O-])=O GTCKPGDAPXUISX-UHFFFAOYSA-N 0.000 description 1
- YBCAZPLXEGKKFM-UHFFFAOYSA-K ruthenium(iii) chloride Chemical compound [Cl-].[Cl-].[Cl-].[Ru+3] YBCAZPLXEGKKFM-UHFFFAOYSA-K 0.000 description 1
- 125000002914 sec-butyl group Chemical group [H]C([H])([H])C([H])([H])C([H])(*)C([H])([H])[H] 0.000 description 1
- 229910000077 silane Inorganic materials 0.000 description 1
- 238000013112 stability test Methods 0.000 description 1
- 125000000542 sulfonic acid group Chemical group 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 125000000999 tert-butyl group Chemical group [H]C([H])([H])C(*)(C([H])([H])[H])C([H])([H])[H] 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 238000002525 ultrasonication Methods 0.000 description 1
- 238000002371 ultraviolet--visible spectrum Methods 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/06—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations
- A61K49/18—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes
- A61K49/1818—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes particles, e.g. uncoated or non-functionalised microparticles or nanoparticles
- A61K49/1887—Agglomerates, clusters, i.e. more than one (super)(para)magnetic microparticle or nanoparticle are aggregated or entrapped in the same maxtrix
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/06—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations
- A61K49/08—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by the carrier
- A61K49/085—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by the carrier conjugated systems
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/06—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations
- A61K49/08—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by the carrier
- A61K49/10—Organic compounds
- A61K49/101—Organic compounds the carrier being a complex-forming compound able to form MRI-active complexes with paramagnetic metals
- A61K49/103—Organic compounds the carrier being a complex-forming compound able to form MRI-active complexes with paramagnetic metals the complex-forming compound being acyclic, e.g. DTPA
- A61K49/105—Organic compounds the carrier being a complex-forming compound able to form MRI-active complexes with paramagnetic metals the complex-forming compound being acyclic, e.g. DTPA the metal complex being Gd-DTPA
-
- C—CHEMISTRY; METALLURGY
- C01—INORGANIC CHEMISTRY
- C01B—NON-METALLIC ELEMENTS; COMPOUNDS THEREOF; METALLOIDS OR COMPOUNDS THEREOF NOT COVERED BY SUBCLASS C01C
- C01B32/00—Carbon; Compounds thereof
- C01B32/20—Graphite
- C01B32/21—After-treatment
- C01B32/23—Oxidation
Definitions
- the present invention relates to the field of medical nanomaterials, and in particular, to a functionalized graphene oxide material having superior magnetic resonance imaging properties, and a method of preparing the same. Background technique
- Graphene is composed of a single atomic layer in which carbon atoms are sp 2 hybridized, and its basic structural unit is the most stable benzene six-membered ring in organic materials.
- graphene has developed rapidly in the field of biomedicine.
- one of the most important derivatives of graphene, graphene oxide, due to its good biocompatibility, high specific surface area, chemically functional modification, can be widely used in biomedical applications for targeted drug delivery. , cell imaging, bioassay, tumor therapy, etc. (Liu, Z.; Robinson, JT; Sun, X.; Dai, HJ Am. Chem. Soc. 2008, 130, 10876. Zhang, L.; Xia, J.
- Magnetic resonance imaging is a non-invasive imaging method that utilizes magnetic resonance phenomena and is imaged by computer technology and two-dimensional image reconstruction. It has the advantages of no ionizing radiation, multi-core, multi-parameter imaging. Medically, it is necessary to obtain accurate diagnostic images and data. However, some of the diseased tissues of the human body sometimes have little difference with the signals of normal tissues, so it is necessary to use contrast agents to help obtain a magnetic resonance image with clear contrast. There are reports that by 2009 global sales of various contrast agents have exceeded $18 billion and are growing globally at a rate of 6-8% per year. According to relevant data, in the domestic Chinese pharmaceutical market, various contrast agents increase at a rate of 25-30% per year.
- the contrast agent on the market is mainly DTPA-Gd, but it has problems such as low relaxation rate, fast metabolism in the body, short retention time and poor stability. Therefore, there is still no contrast agent in the art which has high relaxation rate, slow body metabolism, long retention time and good stability. Summary of the invention
- a contrast agent having a high relaxation rate and good stability and a preparation method thereof.
- a contrast agent having the structure of Formula I:
- R is a ruthenium metal complex formed by complexing a complexing molecule Ra with a ruthenium metal;
- B is a bridging molecule, and the bridging molecule is bonded to the surface of the graphene oxide by a chemical bond, and is connected to the rhodium metal complex by a chemical bond;
- n is greater than 1.
- the functional group is selected from the group consisting of a carboxyl group, a sulfonic acid group, a hydroxyl group, a folic acid group, or the like, or a combination thereof.
- the ruthenium metal complex comprises a complexing molecule and a ruthenium ion complexed with the complexing molecule.
- the graphene oxide comprises modified or unmodified graphene oxide.
- the modified graphene oxide may be graphene oxide having a surface modified with a targeting group or a targeting molecule.
- the targeting molecule comprises a protein, a polypeptide, a nucleic acid, a polysaccharide, a small molecule compound.
- the small molecule compound is selected from the group consisting of folic acid, lactobionic acid, or a combination thereof.
- the targeting molecule comprises an antibody, a ligand, folic acid.
- the bridging molecule is selected from the group consisting of a bifunctional molecule, a polyfunctional molecule, or a combination thereof.
- the bis/polyfunctional molecule is selected from the group consisting of functionalized polyethylene glycol, functionalized polyethyleneimine, functionalized dendrimer, hyaluronic acid, and shell poly Sugar, dextran, polylactic acid
- the complexing molecule is selected from the group consisting of: diethylenetriaminepentaacetic acid (DTPA) or a derivative thereof, 1, 4, 7, 10-tetraazacyclododecane-1, 4, 7, 10-tetra Acetic acid (DOTA) or a derivative thereof, or a combination thereof.
- DTPA diethylenetriaminepentaacetic acid
- DOTA diethylenetriaminepentaacetic acid
- the functional group is selected from the group consisting of hydroxyl, amino, carboxyl, aldehyde, decyl, azide, isocyanate, silane, maleimide, succinimide carbonate Ester, succinimide acetate, succinimide propionate, succinimide succinate, succinimide valerate, or a combination thereof.
- the ester is an ester formed with a substituted or unsubstituted C1-C20 alkyl group or a C3-C20 cycloalkyl group.
- the bridging molecule is a biocompatible molecule. In another preferred embodiment, the bridging molecule is further modified with a group capable of specifically interacting with the cell.
- the diethylenetriaminepentaacetic acid derivative or the 1,4,7,10-tetraazacyclododecane-1,4,7-tetraacetic acid derivative is selected from the group consisting of Group: Diethylenetriaminepentaacetic acid-bisdicarboxamide (DTPA-BDMA), diethylenetriaminepentaacetic acid-bis(isoniazid) (DTPA-BIN), diethylenetriaminepentaacetic acid-bisformamide (DTPA) -BMA), 1, 4, 7, 10-tetraazacyclododecane-1,4,7-triacetic acid (DO3A), 10-(2-hydroxypropyl)-1,4,7, 10- Tetraazacyclododecane-1,4,7-triacetic acid (HPDO3A), 2-methyl-1,4,7,10-tetraazacyclododecane-1,4,7,10- Acetic acid (MCTA), ( ⁇ , ⁇ ', ⁇ " , ⁇ " ' )-tetramethyl-1,4,
- the mass ratio of the bridging molecule to graphene is 1:
- the contrast agent is a contrast agent that can be used for cell imaging.
- the contrast agent is a non-targeting contrast agent.
- the contrast agent has the following properties:
- the relaxation rate is determined as S mM ⁇ S relaxation rate r 1 20 mM ⁇ 1 S ⁇ 1 ; preferably 8 mM- 1 S- 1 relaxation rate r 1 12 mM- 1 S - 1 ; and/or
- the physiological solution is selected from the group consisting of fetal calf serum, DMEM cell culture fluid, or a combination thereof.
- the "non-causation phenomenon” means that the contrast agent of the present invention is mixed with a physiological solution to form a solution having a concentration of 0.25 mM, and after being left at room temperature for 48 hours, a contrast of coagulation occurs. Ratio of agents Lwt%), based on the total weight of the contrast agent.
- a contrast agent according to the first aspect of the invention comprising the steps of:
- A-(B-Ra) n (la) (iii) complexing a graphene oxide having a complex molecule on the surface of the formula la and a phosphonium salt and/or a phosphonium salt solution to obtain a contrast agent of the formula I according to the first aspect of the invention. ;
- A, B, R, n are as defined above;
- Ra is a complex molecule and m is greater than 1.
- n m In another preferred embodiment, n m .
- the steps (ii) and (iii) further comprise a dialysis step.
- the dialysis step is for removing unreacted ruthenium ions, ruthenium metal complexes, bridging molecules, and the like.
- the graphene oxide having a surface-modified bridge molecule is prepared by a method comprising the following steps:
- the method further includes the following features:
- the sulfonium salt is a water-soluble cerium salt, preferably selected from the group consisting of cerium chloride, cerium nitrate, or a combination thereof; the bridging molecule is selected from the group consisting of: functionalized polyethylene glycol, functional group Polyethyleneimine, functionalized dendrimer, hyaluronic acid, chitosan, dextran, polylactic acid-glycolic acid, or a combination thereof.
- a contrast agent composition comprising a diluent and a contrast agent according to the first aspect of the invention.
- the contrast agent is a magnetic resonance imaging contrast agent.
- the contrast agent composition is in the form of a solution.
- the composition further comprises a pharmaceutical component selected from the group consisting of: a therapeutic drug, a tracer molecule, or a combination thereof; and/or
- the composition further comprises a component selected from the group consisting of cerium ions, manganese ions, cerium nanoparticles, manganese nanoparticles, small size ferroferric oxide nanoparticles, or combinations thereof.
- the therapeutic drug is selected from the group consisting of doxorubicin, camptothecin, Iressa, blood root, and the like, or a combination thereof.
- the tracer molecule is selected from the group consisting of fluorescein isothiocyanate, rhodamine, Cy series dyes, AlexaFluor series dyes, doxorubicin, or combinations thereof.
- the small-sized ferroferric oxide nanoparticles have a particle diameter d of ld 5 nm.
- the contrast agent composition according to the first aspect of the invention or the third aspect of the invention is applied to a subject or sample to be contrasted and subjected to contrast.
- the contrast method is non-diagnostic and non-therapeutic.
- the sample is a cell sample.
- the contrast condition is measured at a magnetic field strength of 0.5-20 T, preferably at a magnetic field strength of 0.5-3 T.
- a pharmaceutical composition comprising:
- contrast agent according to the first aspect of the invention, wherein the contrast agent is used as a drug carrier;
- the contrast agent is a targeted drug carrier.
- the drug is combined with the contrast agent by adsorption.
- the pharmaceutically active ingredient is selected from the group consisting of a therapeutic drug, a tracer molecule, or a combination thereof.
- the therapeutic drug is selected from the group consisting of doxorubicin, camptothecin, Iressa, blood root, or a combination thereof.
- the tracer molecule is selected from the group consisting of fluorescein isothiocyanate, rhodamine, Cy series dyes, AlexaFluor series dyes, doxorubicin, or combinations thereof.
- a contrast agent intermediate having the structure la:
- A is graphene oxide or graphene oxide modified by a functional group
- Ra is a complexing molecule, and the complexing molecule can form a complex with a base metal
- B is a bridging molecule, and the bridging molecule is linked to the surface of the graphene by a chemical bond, and is connected to the complexing molecule through a chemical bond;
- n is greater than 1.
- the functionalized modified graphene oxide is selected from the group consisting of carboxyl modified oxygen oxides Motenol, sulfonate-modified graphene oxide, folic acid-modified graphene oxide, or a combination thereof.
- a contrast agent intermediate according to the sixth aspect of the invention for the preparation of a contrast agent according to the first aspect of the invention.
- FIG. 1 is a schematic view showing the preparation of a graphene oxide-based contrast agent of the present invention
- Figure 2 is a diagram showing the enhanced imaging effect of the magnetic resonance imaging contrast agent of the present invention.
- Figure 3 is a graph showing the effect of the magnetic resonance imaging contrast agent of the present invention on cell imaging
- Figure 4a is a UV-Vis spectrum of a magnetic resonance imaging contrast agent of the present invention for loading a drug
- Figure 4b is a graph showing the cell survival rate of a magnetic resonance imaging contrast agent of the present invention acting on a cancer cell after loading the drug.
- a non-specific magnetic resonance imaging contrast agent can be prepared by covalently modifying diethylenetriaminepentaacetate-oxime on graphene oxide.
- the magnetic resonance imaging contrast agent has good imaging effect, wide application range and good stability. Based on the above findings, the inventors completed the present invention.
- bifunctional molecule refers to a molecule having two functional groups within the molecule, said functional group being a group that can covalently link graphene oxide or diethylenetriamine pentaacetic acid.
- the two functional groups may be the same or different.
- substituted means that the hydrogen atom on the group is substituted by one or more substituents selected from the group consisting of halogen, phenyl, C1 to C4 alkyl; wherein the phenyl group includes an unsubstituted phenyl group or A substituted phenyl group having 1 to 3 substituents selected from a hydroxyl group, a C1 to C3 alkyl group, and a halogen.
- C1-C20 alkyl refers to a straight or branched alkyl group having from 1 to 20 carbon atoms, such as methyl, Ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, or the like.
- C3-C20 cycloalkyl refers to a cycloalkyl group having 3 to 20 carbon atoms, such as a cyclopropyl group, a methylcyclopropane group, or the like.
- iridium metal complex refers to a complex molecule and a ruthenium ion complexed with said complex molecule, preferably, said complex is in a chelated form.
- the ruthenium metal complex may be used alone as a contrast agent.
- the ruthenium metal complex is linked to the bridged molecule of the graphene oxide surface by chemical bonding.
- the chemical bond connection mode includes a covalent bond, a coordination bond and the like.
- the ruthenium metal complex is linked to the bridged molecule modified graphene oxide by a non-physical adsorption means.
- Bridge molecule
- bridged molecule refers to a molecule used to attach a graphene surface to a ruthenium metal complex, said bridging molecule being attached to said graphene surface by a chemical bond.
- the bridging molecule is linked to the base metal complex by a chemical bond such as covalent bonding or coordinate bonding.
- the bridging molecule is generally a molecule having two or more functional groups, such as a polyethylene glycol molecule having a functional group at both ends.
- the bridging molecule is a biocompatible molecule.
- the bridging molecule is further modified with a group capable of specifically interacting with cells.
- a group capable of specifically interacting with cells Such as amino acids, peptides, antibodies, nucleic acids, monosaccharides, polysaccharides, vitamins.
- Magnetic resonance imaging contrast agent such as amino acids, peptides, antibodies, nucleic acids, monosaccharides, polysaccharides, vitamins.
- the present invention provides a contrast agent having the following structure:
- A is graphene oxide or graphene oxide modified by a functional group
- R is a ruthenium metal complex formed by complexing a complexing molecule Ra with a ruthenium metal
- B is a bridging molecule, and the bridging molecule is connected to the surface of the graphene oxide by a chemical bond, and is connected to the rhodium metal complex by a chemical bond;
- n is greater than 1.
- the R in another preferred embodiment, may be directly connected to the surface of the graphene, or may be connected to the surface of the graphene through the bridging molecule B, preferably through the bridge.
- Molecular B is attached to the surface of the graphene.
- the graphene oxide may be oxy graphene having a surface modified with a targeting group or a targeting molecule to improve binding of the contrast agent to the target cell line.
- the graphene oxide may also be graphene oxide without modification of a targeting group or a targeting molecule to obtain a non-specific contrast agent.
- the targeting molecule can be any targeting molecule capable of increasing the binding of a contrast agent to a cell line, such as a protein, a polypeptide, a nucleic acid, a polysaccharide, a small molecule compound, and the like.
- the targeting molecule comprises an antibody, a ligand, folic acid.
- the complexing molecule may be any oxidestone which is capable of complexing with cerium ions and capable of being modified with a bridging molecule by a chemical bond such as a coordinate bond or a covalent bond.
- the complexing molecule is selected from the group consisting of: diethylenetriaminepentaacetic acid and derivatives thereof, 1,4,7, 10-tetraazacyclododecane-1, 4, 7, 10- Tetraacetic acid and its derivatives, or a combination thereof.
- the derivative refers to diethylenetriaminepentaacetic acid-bisdic dimethylformamide (DTPA-BDMA;), diethylenetriaminepentaacetic acid-bis(isoniazid) (DTPA-BIN), diethylenetriamine Acetic acid-bisformamide (DTPA-BMA), 1,4,7,10-tetraazacyclododecane-1,4,7-triacetic acid (DO3A), 10-(2-light propyl)-1 ,4,7, 10-tetraazacyclododecane-1,4,7-triacetic acid (HPDO3A), 2-methyl-1,4,7, 10-tetraazacyclododecane-1, 4,7,10-tetraacetic acid (MCTA), ( ⁇ , ⁇ ', ⁇ " , ⁇ " ' )-tetramethyl-1,4,7,10-tetraazacyclododecane-1,4, 7, 10-tetraacetic acid
- the contrast agent can be used alone as a contrast agent or as a contrast agent together with other components.
- the contrast agent further comprises a component selected from the group consisting of cerium ions, manganese ions, cerium nanoparticles, manganese nanoparticles, small-sized ferroferric oxide nanoparticles, or a combination thereof.
- the small size refers to a particle diameter d of l d 5 nm.
- the contrast agent can be used as an intracellular contrast agent for intracellular imaging and detection.
- the contrast agent may also be used in combination with other components, such as a drug loaded together for human or extracorporeal tissue.
- the above drugs may be therapeutic drugs or tracer molecules, such as A. , camptothecin, Iressa, hematoxylin, paclitaxel, fluorescein isothiocyanate, rhodamine, Cy series dyes, AlexaFluor series dyes, etc., or combinations thereof.
- the drug is combined with the contrast agent in an adsorption manner. Since the ruthenium metal complex is chemically bonded to the surface of the modified graphene oxide in the contrast agent, the contrast agent of the present invention can provide a larger The amount of adsorption.
- the drug delivery vehicle has a drug loading rate of 50%, preferably 60%, more preferably 70%.
- the contrast agent can also be used to prepare a pharmaceutical composition, the pharmaceutical composition comprising:
- the contrast agent of the first aspect of the invention as a drug carrier
- the contrast agent is a drug delivery vehicle.
- the drug is combined with the contrast agent by adsorption.
- the drug is not particularly limited and may be doxorubicin, camptothecin, Iressa, blood base, paclitaxel or the like.
- the contrast agent of the present invention can be prepared by the following method:
- A, B, R, n are as defined in the first aspect of the invention.
- Ra is a complex molecule and m is greater than 1.
- n m In another preferred embodiment, n m .
- the steps (ii) and (iii) further comprise a dialysis step.
- the graphene oxide having a surface modified with a bridging molecule can be obtained by a commercially available route or by a conventional method. Method preparation. In another preferred embodiment, the graphene oxide having a surface modified with a bridging molecule can be prepared by a method comprising the following steps:
- the onium salt is a water soluble onium salt, preferably selected from the group consisting of ruthenium chloride, ruthenium nitrate, or combinations thereof.
- the main advantages of the invention include:
- a material suitable for preparing a magnetic resonance imaging contrast agent is provided, and the magnetic resonance imaging contrast agent prepared by the invention has good biocompatibility, and is similar to the existing magnetic resonance imaging contrast agent of the same type. Compared to, it has a better imaging effect.
- the contrast agent of the invention not only significantly improves the contrast effect, but also is suitable for imaging of various human tissues or cells, does not require special processing according to the corresponding tissue or cells, and can be non-specifically Or widely applicable to a variety of different human tissues or cells.
- the contrast agent of the present invention has a long circulation time in the body and is suitable for intracellular imaging.
- the contrast agent of the present invention is easy to load a drug, and has a high drug loading rate of 70% or more.
- the preparation of the magnetic resonance imaging contrast agent is divided into three steps, including preparation of graphene oxide, modification of graphene oxide by polyethylene glycol molecules, covalent attachment of diethylenetriamine pentaacetic acid to complex ruthenium ions, Specific steps are as follows:
- the first step the preparation of graphene oxide:
- the lg flake graphite, 0.5 g potassium persulfate, and 0.5 g phosphorus pentoxide were dissolved in 1.5 mL of concentrated sulfuric acid, heated to 8 CTC for 6 hours, cooled to room temperature, and then washed with water. Neutral, naturally dried into a powder. Add the above powder to 23 mL of concentrated sulfuric acid pre-cooled to 0 ° C, and add 3 g of permanganic acid with stirring. Potassium, keep the temperature below 20 °C. The temperature of the mixture was raised to 35 ° C, and after stirring for 2 h, 46 mL of three times of water was added and stirred for 15 min.
- the second step the preparation of polyethylene oxide molecularly modified graphene oxide:
- the third step is the preparation and application of graphene oxide-diethylenetriaminepentaacetic acid-ruthenium system:
- the polyethylene glycol-modified graphene oxide obtained in the second step reaction was dispersed in an organic phase, 5 mL was added and 0.1 g of diethylenetriamine pentaacetic anhydride was added, and then EDAC 1 mL was added, followed by stirring overnight. The resulting product was dispersed by dialysis into the aqueous phase. GdCl 3 solution was then added, stirred for 4 child removing unreacted GdCl 3, i.e., to obtain the desired contrast agent. Contrast effect test:
- the solution of the contrast medium (concentration: 0.5 mM) and the physiological solution (the fetal bovine serum, and the DMEM cell culture solution, respectively) were mixed in an equal volume ratio to form a solution having a concentration of 0.25 mM, and no significant precipitation was observed by the naked eye.
- concentration of cerium ions remaining on the graphene after being mixed with the fetal bovine serum for 24 hours is 90% or more of the initial concentration of the mixed solution (when the mixing is completed).
- the HepG2 cells were seeded in a cell culture dish, and contrast medium materials of different concentration gradients were added, and after 24 hours of incubation, the cells were digested, and the cells were fixed with agarose solution, and the cell imaging test was performed under the magnetic field strength of 11.7 T ( The result is shown in Figure 3.
- Loaded drug test - Dispensing doxorubicin into an aqueous solution at a concentration of 1 mg/mL adding an equal volume to the contrast solution prepared above, adjusting the pH of the solution, and stirring for about 4 hours in the dark, dialysis removes excess Mycin.
- Step 1 and Step 2 are the same as Embodiment 1.
- the third step is the preparation and application of graphene oxide-(1, 4, 7, 10-tetraazacyclododecane-1, 4, 7, 10-tetraacetic acid)-ruthenium system:
- the polyethylene glycol-modified graphene oxide obtained in the second step reaction is dispersed in the organic phase, and 1 ml of a concentration of 100 mg/mL is added to 5 ml of the polyethylene glycol-modified graphene oxide dispersion. 4, 7, 10-tetraazacyclododecane-1,4,7,10-tetraacetic acid solution, then add EDAC lml, stir overnight, and dialyze off unreacted molecules. Thereafter, a cerium chloride salt solution was added, and after stirring for 4 hours, unreacted Gd ions were removed by dialysis to obtain a desired contrast agent.
- the results show that the contrast agent produced can significantly enhance the contrast effect compared with the commercially available Magenville, which is 2-3 times higher at 1 1. 7T.
- the specific preparation process described above is given by way of example only, and the molecules of the modified graphene oxide mentioned are not limited to the polyethylene glycol, the diethylenetriamine pentaacetic acid, the phosphonium ion mentioned in the above examples, Other molecules such as polyethyleneimine, dendrimer, hyaluronic acid, chitosan, dextran, polylactic acid-glycolic acid, 1, 4, 7, 10-tetraazacyclododecane may be used.
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Veterinary Medicine (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Epidemiology (AREA)
- Radiology & Medical Imaging (AREA)
- Animal Behavior & Ethology (AREA)
- Organic Chemistry (AREA)
- Nanotechnology (AREA)
- Engineering & Computer Science (AREA)
- General Life Sciences & Earth Sciences (AREA)
- Geology (AREA)
- Inorganic Chemistry (AREA)
- Medicinal Chemistry (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Abstract
一种基于氧化石墨烯材料的造影剂及其制备方法,所述造影剂具有式I:A-(B-R) n的结构,其中,A为氧化石墨烯,R为钆金属络合物,B为桥联分子,且A、B、R之间通过化学键相互结合;该造影剂具有驰豫高、稳定性好、非特异性等特征。
Description
基于氧化石墨烯材料的造影剂及其制备方法
技术领域
本发明涉及医用纳米材料领域, 具体地, 本发明涉及一种具有优越磁共振造 影性能的功能化氧化石墨烯材料, 及其制备方法。 背景技术
石墨烯由碳原子以 sp2杂化连接的单原子层构成, 其基本结构单元为有机材料中 最稳定的苯六元环。近两年来石墨烯在生物医学领域的研究发展迅速。特别是石墨烯 最重要的衍生物之一, 氧化石墨烯, 由于其良好的生物相容性, 超高的比表面积, 能 够化学功能化修饰,在生物医学上可广泛应用于靶向药物输运、细胞成像、生物检测、 肿瘤治疗等方面(Liu, Z.; Robinson, J. T.; Sun, X.; Dai, H. J. Am. Chem. Soc. 2008, 130, 10876. Zhang, L.; Xia, J.; Zhao, Q.; Liu, L.; Zhang, Z. Small. 2010, 6, 537. Gon alves, G.; Vila, M.; Portoles, M. T.; Vallet-Regi, M.; Gracio, J.; Marques, P. A. Adv. Healthcare Mater. 2013, 2, 1072.) 0
磁共振成像是利用磁共振现象, 借助计算机技术和二维图像重建方法进行成 像, 是一种无创的影像手段。 其具有无电离辐射、 可实现多核、 多参数成像等优 点。 医学临床上需要得到精确的诊断学图像和数据, 但人体的某些病变组织有时 与正常组织之间的信号差异不大, 故需要使用造影剂帮助得到对比度清晰的磁共 振图像。有报告显示至 2009年全球各类造影剂销售额已超过 180亿美元, 且以每 年 6-8%的速度在全球范围内增长。 根据有关数据显示, 在中国国内医药市场上, 各种造影剂每年以 25-30%的速率递增。
目前市场上的造影剂主要是 DTPA-Gd, 但其存在着弛豫率偏低, 在体内代谢 快, 存留时间短, 稳定性差等问题。 因此, 本领域尚缺乏一种弛豫率高, 体内代 谢慢, 存留时间长, 稳定性好的造影剂。 发明内容
本发明的目的是提供一种弛豫率高, 稳定性好的造影剂及其制备方法。 在本发明的第一方面, 提供了一种造影剂, 所述的造影剂具有式 I结构:
A-(B-R)n (I)
式中, A为氧化石墨烯, 或被功能化基团修饰的氧化石墨烯;
R为钆金属络合物, 所述钆金属络合物由络合分子 Ra与钆金属络合形成;
B为桥联分子, 所述的桥联分子通过化学键与所述的氧化石墨烯表面相连, 并通 过化学键与所述的钆金属络合物相连;
"-"表示化学键;
n大于 1。
在另一优选例中 , 所述的功能化基团选自下组: 羧基、 磺酸基、 羟基、 叶酸等基 团或分子, 或其组合
在另一优选例中 ,所述的钆金属络合物包括络合分子和与所述的络合分子络合的 钆离子。
在另一优选例中 , 所述的氧化石墨烯包括经修饰的或未经修饰的氧化石墨烯。 在另一优选例中,所述的经修饰的氧化石墨烯可以是表面修饰有靶向基团或靶向 分子的氧化石墨烯。
在另一优选例中 , 所述的靶向分子包括蛋白、 多肽、核酸、 多糖、小分子化合物。 在另一优选例中, 所述的小分子化合物选自下组: 叶酸、 乳糖酸, 或其组合。 在另一优选例中, 所述的靶向分子包括抗体、 配体、 叶酸。
在另一优选例中 , 所述的桥联分子选自下组: 双官能团分子、 多官能团分子, 或 其组合。
在另一优选例中 , 所述的双 /多官能团分子选自下组: 官能团化的聚乙二醇、 官 能团化的聚乙烯亚胺、 官能团化的树状大分子、 透明质酸、 壳聚糖、 葡聚糖、 聚乳酸
-羟基乙酸, 或其组合; 和 /或
所述的络合分子选自下组: 二乙三胺五乙酸 (DTPA)或其衍生物, 1, 4, 7, 10-四氮 杂环十二烷 -1, 4, 7, 10-四乙酸(D0TA)或其衍生物, 或其组合。
在另一优选例中, 所述的官能团选自下组: 羟基、 氨基、 羧基、 醛基、 巯基、 叠 氮基、 异氰酸基、 硅烷基、 马来酰亚胺、 琥珀酰亚胺碳酸酯、 琥珀酰亚胺乙酸酯、 琥珀酰亚胺丙酸酯、 琥珀酰亚胺琥珀酸酯、 琥珀酰亚胺戊酸酯, 或其组合。
在另一优选例中, 所述酯为与取代或未取代的 C1-C20烷基或 C3-C20的环烷基 所形成的酯。
在另一优选例中, 所述的桥联分子是生物相容性分子。
在另一优选例中,所述的桥联分子上还修饰有能够特异性地与细胞相互作用的基 团。
在另一优选例中, 所述的二乙三胺五乙酸衍生物或 1, 4, 7, 10-四氮杂环十二烷 -1, 4, 7, 10-四乙酸衍生物选自下组: 二乙三胺五乙酸 -双二甲酰胺 (DTPA-BDMA)、 二乙三胺五乙酸-双 (异烟肼) (DTPA-BIN)、二乙三胺五乙酸-双甲酰胺 (DTPA-BMA)、 1, 4, 7, 10- 四氮杂环十二烷 -1, 4, 7- 三乙酸 (DO3A)、 10-(2-羟丙基 )-1,4,7, 10- 四氮杂 环十二烷 -1,4,7- 三乙酸 (HPDO3A)、 2- 甲基 -1,4,7,10- 四氮杂环十二烷 -1,4,7,10- 四乙 酸 (MCTA)、 (α,α',α " ,α " ' )- 四甲基 -1,4,7,10- 四氮杂环十二烷 -1,4,7,10- 四乙酸 (DOTMA)等, 或其组合。
在另一优选例中, 在所述的造影剂中, 所述的桥联分子与石墨烯的质量比为 1 :
0.2〜5。
在另一优选例中, 所述的造影剂是可用于细胞成像的造影剂。
在另一优选例中, 所述的造影剂是非靶向性造影剂。
在另一优选例中, 所述的造影剂具有以下性能:
在 11.7 T磁场强度下, 弛豫率 测定为 S mM^S 弛豫率r1 20 mM·1S·1 ; 较佳 地为 8 mM-1S-1 弛豫率r1 12 mM-1S-1 ; 禾口 /或
在生理性溶液中无聚沉现象出现。
在另一优选例中, 所述的生理性溶液选自下组: 胎牛血清、 DMEM细胞培养液, 或其组合。
在另一优选例中, 所述的 "无聚沉现象"指, 将本发明造影剂与生理性溶液混合 后形成浓度为 0.25 mM的溶液, 在室温下放置 48小时后, 发生聚沉的造影剂比例
lwt%), 按造影剂的总重量计。
本发明的第二方面, 提供了一种如本发明第一方面所述的造影剂的制备方法, 所 述的方法包括以下步骤:
(i) 提供一表面连接有桥联分子的氧化石墨烯 A-(B)m, 其中所述桥联分子 B通过 化学键连于氧化石墨烯 A的表面;
(ϋ) 用络合分子 Ra与所述的表面连接有桥联分子的氧化石墨烯 - ^进行反应, 形成表面连接有络合分子的、 式 la所示的氧化石墨烯;
A-(B-Ra)n (la)
(iii) 将式 la所示的表面连接有络合分子的氧化石墨烯与钆盐和 /或钆盐溶液进行 络合反应, 得到如本发明第一方面所述的式 I所示的造影剂;
其中, A、 B、 R、 n的定义如上文所述;
Ra为络合分子, m大于 1。
在另一优选例中, n m。
在另一优选例中, 对所述步骤 (ii)和 (iii)还包括透析步骤。
在另一优选例中, 所述的透析步骤用于去除未反应的钆离子、 钆金属络合物、 桥 联分子等反应物。
在另一优选例中,所述的表面修饰有桥联分子的氧化石墨烯通过包括以下步骤的 方法制备:
(a) 提供一氧化石墨烯;
(b) 将桥联分子与所述的氧化石墨烯反应, 得到表面修饰有桥联分子的氧化石墨 烯。
在另一优选例中, 所述方法还包括以下特征:
所述的钆盐为水溶性钆盐, 较佳地选自下组: 氯化钆、 硝酸钆, 或其组合; 所述的桥联分子选自下组: 官能团化的聚乙二醇、 官能团化的聚乙烯亚胺、 官能 团化的树状大分子、 透明质酸、 壳聚糖、 葡聚糖、 聚乳酸 -羟基乙酸, 或其组合。
本发明的第三方面, 提供了一种造影剂组合物, 所述的造影剂包括稀释剂以及如 本发明第一方面所述的造影剂。
在另一优选例中, 所述的造影剂是磁共振成像造影剂。
在另一优选例中, 所述的造影剂组合物为溶液形式。
在另一优选例中, 所述的组合物还包括选自下组的药物组分: 治疗性药物、 示踪 性分子, 或其组合; 和 /或
所述组合物还包括选自下组的组分: 钆离子、 锰离子、 钆纳米粒子、 锰纳米粒 子、 小尺寸四氧化三铁纳米粒子, 或其组合。
在另一优选例中, 所述的治疗性药物选自下组: 阿霉素、 喜树碱、 易瑞沙、 血根 碱等, 或其组合。
在另一优选例中, 所述的示踪性分子选自下组: 异硫氰酸荧光素、 罗丹明、 Cy 系列染料、 AlexaFluor系列染料、 阿霉素, 或其组合。
在另一优选例中, 所述的小尺寸四氧化三铁纳米粒子的粒径 d为 l d 5nm。 本发明的第四方面, 提供了一种造影方法, 所述方法包括步骤:
将本发明第一方面或本发明第三方面所述的造影剂组合物施用于待造影的对象 或样品, 并进行造影。
在另一优选例中, 所述的造影方法是非诊断和非治疗性的。
在另一优选例中, 所述的样品是细胞样品。
在另一优选例中, 所述造影条件为在 0.5-20T磁场强度下测定, 优选在 0.5-3T磁 场强度下测定。
本发明的第五方面, 提供了一种药物组合物, 所述的药物组合物包括:
©如本发明第一方面所述的造影剂, 所述造影剂作为药物载体;
(ii)负载于所述造影剂上的药物活性成分; 和 (iii)药学上可接受的其他载体。
在另一优选例中, 所述的造影剂为靶向性药物运载体。
在另一优选例中, 所述的药物通过吸附方式与所述的造影剂结合。
在另一优选例中, 所述的药物活性成分选自下组: 治疗性药物、 示踪性分子, 或 其组合。
在另一优选例中, 所述的治疗性药物选自下组: 阿霉素、 喜树碱、 易瑞沙、 血根 碱, 或其组合。
在另一优选例中, 所述的示踪性分子选自下组: 异硫氰酸荧光素、 罗丹明、 Cy 系列染料、 AlexaFluor系列染料、 阿霉素, 或其组合。
本发明的第六方面, 提供了一种造影剂中间体, 所述的造影剂中间体具有式 la 结构:
A-(B-Ra)n (la)
式中, A为氧化石墨烯, 或被功能化基团修饰的氧化石墨烯;
Ra为络合分子, 所述络合分子可与钆金属形成络合物;
B为桥联分子, 所述的桥联分子通过化学键与所述的石墨烯表面相连, 并通过化 学键与所述的络合分子相连;
"-"表示化学键;
n大于 1。
在另一优选例中, 所述的功能化修饰的氧化石墨烯选自下组: 羧基修饰的氧化石
墨烯、 磺酸基修饰的氧化石墨烯、 叶酸修饰的氧化石墨烯, 或其组合。
本发明的第七方面,提供了一种本发明第六方面所述的造影剂中间体用于制备如 本发明第一方面所述的造影剂的用途。 应理解, 在本发明范围内中, 本发明的上述各技术特征和在下文 (;如实施例) 中具体描述的各技术特征之间都可以互相组合,从而构成新的或优选的技术方案。 限于篇幅, 在此不再一一累述。 附图说明
图 1是本发明的基于氧化石墨烯的造影剂的制备示意图;
图 2是本发明的磁共振成像造影剂增强成像效果图;
图 3是本发明的磁共振成像造影剂用于细胞成像效果图;
图 4a是本发明的磁共振成像造影剂用于负载药物的紫外-可见光谱图; 图 4b是本发明的磁共振成像造影剂负载药物后作用于癌细胞的细胞存活率 图。 具体实施方式
本发明人经过长期而深入的研究, 意外地发现, 通过在氧化石墨烯上共价修 饰二乙三胺五乙酸 -钆, 可以制备得到非特异性的磁共振成像造影剂。 所述的磁共 振成像造影剂成像效果好, 适用范围广, 稳定性好。 基于上述发现, 发明人完成 了本发明。 术语
如本文所用, 术语 "双官能团分子" 指分子内具有两个功能性基团的分子, 所述的功能性基团是可以共价连接氧化石墨烯或二乙三胺五乙酸的基团。 所述的 两个功能性基团可相同或不同。
术语 "取代" 是指基团上的氢原子被一个或多个选自下组的取代基取代: 卤 素、 苯基、 C1〜C4烷基; 其中所述的苯基包括未取代的苯基或具有 1-3个选自羟 基、 C1〜C3烷基、 卤素的取代基的取代苯基。
术语 " C1-C20烷基" 指具有 1-20个碳原子的直链或支链烷基, 例如甲基、
乙基、 丙基、 异丙基、 丁基、 异丁基、 仲丁基、 叔丁基、 或类似基团。 术语 " C3-C20环烷基" 指具有 3-20个碳原子的环烷基, 例如环丙烷基、 甲 基环丙烷基、 或类似基团。 礼金属络合物
如本文所用, 术语 "钆金属络合物"指络合分子以及与所述的络合分子络合的钆 离子, 较佳地, 所述的络合是螯合形式。
所述的钆金属络合物可以单独作为造影剂使用, 在本发明中, 所述的钆金属络合 物通过化学键连接方式与氧化石墨烯表面修饰的桥联分子连接。其中, 所述的化学键 连接方式包括共价键, 配位键等。
特别地,所述的钆金属络合物通过非物理吸附方式与桥联分子修饰的氧化石墨烯 连接。 桥联分子
如本文所用, 术语 "桥联分子"指用于连接石墨烯表面与钆金属络合物的分子, 所述的桥联分子通过化学键与所述的石墨烯表面相连。
特别地, 所述的桥联分子通过共价结合、配位键结合等化学键连接方式与钆金属 络合物相连。
所述的桥联分子一般为具有两个或两个以上功能性基团的分子,如双端具有官能 团的聚乙二醇分子等。
在另一优选例中, 所述的桥联分子是生物相容性分子。
在另一优选例中,所述的桥联分子上还修饰有能够特异性地与细胞相互作用的基 团。 如氨基酸、 肽、 抗体、 核酸、 单糖、 多糖、 维生素。 磁共振成像造影剂
本发明提供了一种造影剂, 所述的造影剂具有以下结构:
A-(B-R)n (I)
式中, A为氧化石墨烯, 或被功能化基团修饰的氧化石墨烯;
R为钆金属络合物, 所述钆金属络合物由络合分子 Ra与钆金属络合形成;
B为桥联分子, 所述的桥联分子通过化学键与所述的氧化石墨烯表面相连, 并通 过化学键与所述的钆金属络合物相连;
"-"表示化学键;
n大于 1。
在另一优选例中, 在所述的造影剂中, 所述的 R可以直接与石墨烯表面相连, 也可以通过所述的桥联分子 B与石墨烯表面连接, 优选通过所述的桥联分子 B与石 墨烯表面连接。
在另一优选例中,所述的氧化石墨烯可以是表面修饰有靶向基团或靶向分子的氧 化石墨烯, 以改善造影剂与目标细胞系的结合。所述的氧化石墨烯也可以是未经修饰 有靶向基团或靶向分子的氧化石墨烯, 以得到非特异性的造影剂。
所述的靶向分子可以是任何能够增加造影剂和细胞系的结合的靶向分子, 如蛋 白、 多肽、 核酸、 多糖、 小分子化合物等。
在另一优选例中, 所述的靶向分子包括抗体、 配体、 叶酸。
所述的络合分子可以是任意能够与钆离子络合,且能够与桥联分子修饰的氧化石 墨烯通过化学键 (如配位键或共价键)相连。 较佳地, 所述的络合分子选自下组: 二乙 三胺五乙酸以及其衍生物、 1,4,7, 10-四氮杂环十二烷 -1, 4, 7, 10-四乙酸以及其衍 生物,或其组合。其中,所述的衍生物指二乙三胺五乙酸 -双二甲酰胺 (DTPA-BDMA;)、 二乙三胺五乙酸-双 (异烟肼 )(DTPA-BIN)、 二乙三胺五乙酸-双甲酰胺 (DTPA-BMA)、 1,4,7,10- 四氮杂环十二烷 -1,4,7- 三乙酸 (DO3A)、 10-(2- 轻丙基 )-1,4,7, 10- 四氮杂环 十二烷 -1,4,7- 三乙酸 (HPDO3A)、 2- 甲基 -1,4,7, 10- 四氮杂环十二烷 -1,4,7,10- 四乙酸 (MCTA)、 (α,α',α " ,α " ' )- 四甲基 -1,4,7,10- 四氮杂环十二烷 -1,4,7, 10- 四乙酸
(DOTMA)等, 或其组合。
所述的造影剂可以单独作为造影剂使用, 也可以和其他组分共同作为造影剂使 用。 在另一优选例中, 所述的造影剂还包括选自下组的组分: 钆离子、 锰离子、 钆 纳米粒子、 锰纳米粒子、 小尺寸四氧化三铁纳米粒子,或其组合。 其中, 所述的小 尺寸指粒径 d为 l d 5 nm。
特别地, 所述的造影剂可以用作细胞内造影剂, 用于进行细胞内成像和检测。 在另一优选例中, 所述的造影剂还可以与其他组分共同使用, 如负载药物共 同用于人体或体外组织等。 上述的药物可以是治疗性药物或示踪性分子等, 如阿霉
素、 喜树碱、 易瑞沙、 血根碱、 紫杉醇、 异硫氰酸荧光素、 罗丹明、 Cy系列染料、 AlexaFluor系列染料等, 或其组合。
所述的药物与所述的造影剂以吸附方式结合, 由于所述造影剂中, 钆金属络合物 以化学键方式与修饰的氧化石墨烯表面结合, 因此, 本发明的造影剂可以提供较大的 吸附量。在另一优选例中, 所述的药物递送载体的载药率为 50%, 较佳地为 60%, 更佳地为 70%。
在另一优选例中, 所述的造影剂还可以用于制备药物组合物, 所述的药物组 合物包括:
«如本发明第一方面所述的造影剂, 所述造影剂作为药物载体;
(ii)负载于所述造影剂上的药物活性成分; 和
(iii)药学上可接受的其他载体。
在另一优选例中, 所述的造影剂为药物递送载体。
在另一优选例中, 所述的药物通过吸附方式与所述的造影剂结合。
所述的药物没有特别限制, 可以是阿霉素、喜树碱、 易瑞沙、血根碱、紫杉醇等。 造影剂的制备
本发明所述的造影剂可以通过以下方法制备:
(i) 提供一表面连接有桥联分子的氧化石墨烯 A-(B)m, 其中所述桥联分子 B通过 化学键连于氧化石墨烯 A的表面;
(ii) 用络合分子 Ra与所述的表面连接有桥联分子的氧化石墨烯 A-(B)m进行反 应, 形成表面连接有络合分子的、 式 la所示的氧化石墨烯;
A-(B-Ra)n (la)
(iii) 将式 la所示的表面连接有络合分子的氧化石墨烯与钆盐和 /或钆盐溶液进行 络合反应, 得到如本发明第一方面所述的式 I所示的造影剂;
其中, A、 B、 R、 n的定义如本发明第一方面所述;
Ra为络合分子, m大于 1。
在另一优选例中, n m。
在另一优选例中, 对所述步骤 (ii)和 (iii)还包括透析步骤。
所述的表面修饰有桥联分子的氧化石墨烯可以通过市售途径获得,或通过常规方
法制备。在另一优选例中, 所述的表面修饰有桥联分子的氧化石墨烯可以通过包括以 下步骤的方法制备:
(a) 提供一氧化石墨烯;
(b) 用桥联分子与所述的氧化石墨烯反应, 得到表面修饰有桥联分子的氧化石墨 烯。
所述的钆盐为水溶性钆盐, 较佳地选自下组: 氯化钆、 硝酸钆, 或其组合。 本发明的主要优点包括:
(1) 提供了一种适用于制备磁共振成像造影剂的材料, 本发明所制备的磁共 振成像造影剂具有良好的生物相容性, 且与现有的同类型磁共振成像造影剂马根 维显相比, 具有更好的成像效果。
(2) 相较于现有技术, 本发明的造影剂不仅显著改善了造影效果, 而且适用 于多种人体组织或细胞的成像, 不需要根据相应的组织或细胞进行特殊加工, 能 够非特异性地或广泛地适用于多种不同的人体组织或细胞。
(3) 本发明的造影剂体内循环时间长, 并适合用于进行细胞内成像。
(4) 本发明的造影剂易于负载药物, 且载药率高, 可达到 70%或更高。
下面结合具体实施例, 进一步阐述本发明。 应理解, 这些实施例仅用于说明本 发明而不用于限制本发明的范围。 下列实施例中未注明具体条件的实验方法, 通 常按照常规条件, 或按照制造厂商所建议的条件。 除非另外说明, 否则百分比和 份数按重量计算。 实施例 1 磁共振成像造影剂的制备
本实施例中, 磁共振成像造影剂的制备分为三个步骤, 包括氧化石墨烯的制 备, 聚乙二醇分子修饰氧化石墨烯, 共价连接二乙三胺五乙酸以络合钆离子, 具 体步骤如下:
第一步, 氧化石墨烯的制备:
将 l g片状石墨、 0. 5 g过硫酸钾、 0. 5 g五氧化二磷溶于 1. 5 mL浓硫酸中, 加热至 8CTC反应 6小时, 冷却到室温后, 再用水将其洗成中性, 自然干燥成粉末。 将上述粉末加入到 23 mL事先预冷至 0°C的浓硫酸中, 边搅拌边加入 3 g高锰酸
钾, 保持温度在 20 °C以下。 将混合物的温度升至 35 °C, 搅拌 2 h后, 加入 46 mL 三次水, 搅拌 15 min。 为中止氧化反应, 加入 140 mL三次水和 2 mL浓度为 30% 的过氧化氢。 将混合物用 250mL 10%的盐酸离心清洗, 以除去溶液中的金属离子。 将上述氧化石墨用超声 2 h, 10000转 /分钟离心 30 min到 1 h, 上清液即为氧化 石墨烯, 进一步超声可制备出纳米尺度的氧化石墨烯。
第二步, 聚乙二醇分子修饰的氧化石墨烯的制备:
取第一步反应中得到的氧化石墨烯 5 mL预先超声半小时, 充分分散后, 调 pH值约为 8,加入聚乙二醇分子(PEG)水溶液 lmL,在室温搅拌下,加入 EDAC 0. 15 mL, 继续搅拌, 再次加入 EDAC 0. 35 mL , 搅拌过夜。 随后透析得到 PEG修饰的氧 化石墨烯。
第三步, 氧化石墨烯-二乙三胺五乙酸-钆体系的制备及应用:
将第二步反应中得到的聚乙二醇修饰的氧化石墨烯分散到有机相中, 取 5 mL 并加入二乙三胺五乙酸酸酐 0. 1 g, 再加入 EDAC l mL, 之后搅拌过夜。 得到的产 物经透析分散到水相中。随后加入 GdCl3溶液,搅拌 4小时候除去未反应的 GdCl3, 即得到所需的造影剂。 造影效果测试:
该造影剂的制备示意图如图 1所示, 其与市售马根维显相比, 能显著增强造 影效果(图 2所示), 在 11. 7 T场强下约 2. 4倍, 为 10. 8 m TS— 稳定性测试:
该造影剂水溶液(浓度为 0.5 mM)与生理性溶液 (分别为胎牛血清、 和 DMEM 细胞培养液) 等体积比混合后, 形成浓度为 0. 25 mM的溶液, 肉眼均未见明显沉 淀。 其中, 与胎牛血清混合 24h后留存在石墨烯上的钆离子浓度为该混合液的初 始浓度(完成混合时)的 90%以上。 细胞内成像测试:
将 HepG2细胞接种于细胞培养皿内, 分别加入不同浓度梯度的造影剂材料, 共同孵育 24小时后, 消化收集细胞, 并用琼脂糖溶液固定细胞, 在 11. 7 T磁场 强度下进行细胞成像测试(结果如图 3所示)。
负载药物测试- 将阿霉素配置成浓度为 1 mg/mL的水溶液, 将其等体积加入上述制备的造影 剂溶液中, 调节溶液 pH值, 避光搅拌约 4小时后, 透析除去多余的阿霉素。 结果 显示, 本发明的造影剂能够负载阿霉素(图 4a所示), 载药率可达到 70%, 并能有 效地杀伤癌细胞(图 4b所示)。 实施例 2 磁共振成像造影剂的制备
步骤一、 步骤二同实施例 1。
第三步,氧化石墨烯- (1, 4, 7, 10-四氮杂环十二烷 -1, 4, 7, 10-四乙酸) -钆体系 的制备及应用:
将第二步反应中得到的聚乙二醇修饰的氧化石墨烯分散到有机相中, 在 5ml 聚乙二醇修饰的氧化石墨烯分散液中, 加入 1 ml浓度为 100 mg/mL的 1, 4, 7, 10- 四氮杂环十二烷 -1,4, 7, 10-四乙酸溶液, 再加入 EDAC lml , 搅拌过夜, 透析出去 未反应的分子。 之后加入氯化钆盐溶液, 搅拌 4小时后, 透析除去未反应的 Gd 离子, 即得到所需的造影剂。
结果显示,制得的造影剂与市售马根维显相比能显著增强造影效果,在 1 1. 7T 下提高 2-3倍。 上述具体的制备工艺仅是作为示例给出, 其中所提到的修饰氧化石墨烯的分 子不仅仅局限于上述例子中所提到的聚乙二醇、 二乙三胺五乙酸、 钆离子, 也可 以选用其它分子, 如聚乙烯亚胺、 树状大分子、 透明质酸、 壳聚糖、 葡聚糖、 聚乳 酸-羟基乙酸、. 1, 4, 7, 10-四氮杂环十二烷 -1, 4, 7, 10-四乙酸、锰离子、锰纳米粒子、 小尺寸四氧化三铁纳米粒子或其组合。 在本发明提及的所有文献都在本申请中引用作为参考, 就如同每一篇文献被 单独引用作为参考那样。 此外应理解, 在阅读了本发明的上述讲授内容之后, 本 领域技术人员可以对本发明作各种改动或修改, 这些等价形式同样落于本申请所 附权利要求书所限定的范围。
Claims
1、 一种造影剂, 其特征在于, 所述的造影剂具有式 I结构:
A-(B-R)n (I)
式中, A为氧化石墨烯, 或被功能化基团修饰的氧化石墨烯;
R为钆金属络合物, 所述钆金属络合物由络合分子 Ra与钆金属络合形成;
B为桥联分子, 所述的桥联分子通过化学键与所述的氧化石墨烯表面相连, 并通 过化学键与所述的钆金属络合物相连;
"-"表示化学键;
n大于 1。
2、 如权利要求 1所述的造影剂, 其特征在于, 所述的桥联分子选自下组: 双官 能团分子、 多官能团分子, 或其组合。
3、 如权利要求 1所述的造影剂, 其特征在于, 所述的经修饰的氧化石墨烯是表 面修饰有靶向基团或靶向分子的氧化石墨烯; 较佳地, 所述的靶向分子选自下组: 蛋 白、 多肽、 核酸、 多糖、 小分子化合物。
4、如权利要求 1所述的造影剂, 其特征在于,所述的双 /多官能团分子选自下组: 官能团化的聚乙二醇、 官能团化的聚乙烯亚胺、 官能团化的树状大分子、 透明质酸、 壳聚糖、 葡聚糖、 聚乳酸 -羟基乙酸, 或其组合; 和 /或
所述的络合分子选自下组: 二乙三胺五乙酸 (DTPA)或其衍生物, 1, 4, 7, 10-四氮 杂环十二烷 -1, 4, 7, 10-四乙酸(D0TA)或其衍生物, 或其组合。
5、 如权利要求 1所述的造影剂的制备方法, 其特征在于, 所述的方法包括以下 步骤:
(i) 提供一表面连接有桥联分子的氧化石墨烯 A-(B)m,其中所述桥联分子 B通过 化学键连于氧化石墨烯 A的表面;
(ii) 用络合分子 Ra与所述的表面连接有桥联分子的氧化石墨烯 A-(B)m进行反 应, 形成表面连接有络合分子的、 式 la所示的氧化石墨烯;
A-(B-Ra)n (la)
(iii) 将式 la所示的表面连接有络合分子的氧化石墨烯与钆盐和 /或钆盐溶液进行 络合反应, 得到如权利要求 1所述的式 I所示的造影剂;
其中, A、 B、 R、 n的定义如权利要求 1所述;
Ra为络合分子, m大于 1。
6、 如权利要求 5所述的制备方法, 其特征在于, 所述的表面修饰有桥联分子的 氧化石墨烯通过包括以下步骤的方法制备:
(a) 提供一氧化石墨烯;
(b) 将桥联分子与所述的氧化石墨烯反应, 得到表面修饰有桥联分子的氧化石墨 烯。
7、 如权利要求 5所述的制备方法, 其特征在于, 还包括以下特征:
所述的钆盐为水溶性钆盐, 较佳地选自下组: 氯化钆、 硝酸钆, 或其组合; 所述的桥联分子选自下组: 官能团化的聚乙二醇、 官能团化的聚乙烯亚胺、 官能 团化的树状大分子、 透明质酸、 壳聚糖、 葡聚糖、 聚乳酸 -羟基乙酸, 或其组合。
8、 一种造影剂组合物, 其特征在于, 所述的造影剂包括稀释剂以及权利要求 1 所述的造影剂。
9、 如权利要求 8所述的造影剂组合物, 其特征在于, 所述的组合物还包括选自 下组的药物组分: 治疗性药物、 示踪性分子, 或其组合; 和 /或
所述组合物还包括选自下组的组分: 钆离子、 锰离子、 钆纳米粒子、 锰纳米粒 子、 小尺寸四氧化三铁纳米粒子, 或其组合。
10、 如权利要求 9所述的造影剂组合物, 其特征在于, 所述的治疗性药物选自下 组: 阿霉素、 喜树碱、 易瑞沙、 血根碱等, 或其组合; 和 /或
所述的示踪性分子选自下组: 异硫氰酸荧光素、 罗丹明、 Cy系列染料、 AlexaFluor系列染料、 阿霉素, 或其组合。
11、 一种造影方法, 其特征在于, 包括步骤:
将权利要求 1或权利要求 6所述的造影剂组合物施用于待造影的对象或样品,并 进行造影。
12、 一种药物组合物, 其特征在于, 所述的药物组合物包括:
©如权利要求 1所述的造影剂, 所述造影剂作为药物载体;
(ii)负载于所述造影剂上的药物活性成分; 和 (iii)药学上可接受的其他载体。
13、 如权利要求 12所述的药物组合物, 其特征在于, 所述的药物活性成分选自 下组: 治疗性药物、 示踪性分子, 或其组合。
14、 一种造影剂中间体, 其特征在于, 所述的造影剂具有式 la结构:
A-(B-Ra)n (la)
式中, A为氧化石墨烯, 或被功能化基团修饰的氧化石墨烯;
Ra为络合分子, 所述络合分子可与钆金属形成络合物;
B为桥联分子, 所述的桥联分子通过化学键与所述的石墨烯表面相连, 并通过化 学键与所述的络合分子相连;
"-"表示化学键;
n大于 1。
15、 一种权利要求 14所述的造影剂中间体的用途, 其特征在于, 用于制备权利 要求 1所述的造影剂。
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201310435341.7A CN104436221B (zh) | 2013-09-23 | 2013-09-23 | 基于氧化石墨烯材料的造影剂及其制备方法 |
CN201310435341.7 | 2013-09-23 |
Publications (1)
Publication Number | Publication Date |
---|---|
WO2015039555A1 true WO2015039555A1 (zh) | 2015-03-26 |
Family
ID=52688218
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/CN2014/085611 WO2015039555A1 (zh) | 2013-09-23 | 2014-08-29 | 基于氧化石墨烯材料的造影剂及其制备方法 |
Country Status (2)
Country | Link |
---|---|
CN (1) | CN104436221B (zh) |
WO (1) | WO2015039555A1 (zh) |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2017066268A1 (en) * | 2015-10-12 | 2017-04-20 | Ohio University | Supercapacitor-based biosensors and methods of detecting a biomarker |
CN108042809A (zh) * | 2018-02-01 | 2018-05-18 | 南京林业大学 | 一种抑菌功能化氧化石墨烯的制备方法 |
Families Citing this family (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN104815337A (zh) * | 2015-05-14 | 2015-08-05 | 临沂大学 | 一种peg修饰的磁性氧化石墨烯的制备及其抗肿瘤活性 |
CN105797174B (zh) * | 2016-01-22 | 2019-01-11 | 复旦大学附属肿瘤医院 | 一种基于纳米氧化石墨烯的磁共振成像造影剂及其制备方法 |
CN107343963A (zh) * | 2016-05-04 | 2017-11-14 | 复旦大学附属肿瘤医院 | 一种用于SPECT显像的99mTc标记纳米探针及其制备方法 |
CN107625964A (zh) * | 2017-08-24 | 2018-01-26 | 杨蕾 | 一种改性氧化石墨烯载药缓释体系的制备方法 |
CN108159417A (zh) * | 2018-01-26 | 2018-06-15 | 北京欧美中科学技术研究院 | 一种石墨烯药物载体材料的制备方法 |
CN111517316B (zh) * | 2020-05-07 | 2022-05-17 | 中国科学院高能物理研究所 | 稀土元素标记氧化石墨烯纳米片、其制备方法及应用 |
CN113234442B (zh) * | 2021-05-11 | 2022-06-24 | 中国科学院上海微系统与信息技术研究所 | 一种顺磁性手性石墨烯量子点及其制备方法和用途 |
CN117550845B (zh) * | 2024-01-11 | 2024-04-12 | 太原理工大学 | 高介电常数水溶性rGO水泥基复合材料的制备方法 |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102397563A (zh) * | 2010-09-16 | 2012-04-04 | 同济大学 | 一种磁共振成像造影剂用石墨烯纳米载体的制备方法 |
CN102657872A (zh) * | 2012-05-05 | 2012-09-12 | 上海师范大学 | 氧化石墨烯/PAMAM/DTPA-Gd/PSCA抗体多功能材料及其制备方法和应用 |
US20130079503A1 (en) * | 2011-09-08 | 2013-03-28 | Research & Business Foundation Sungkyunkwan University | Go-gd-dtpa complex, preparation method thereof, and mri contrast agent comprising the same |
Family Cites Families (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101670108A (zh) * | 2009-08-13 | 2010-03-17 | 苏州纳米技术与纳米仿生研究所 | 基于纳米氧化石墨烯的载药体系 |
-
2013
- 2013-09-23 CN CN201310435341.7A patent/CN104436221B/zh active Active
-
2014
- 2014-08-29 WO PCT/CN2014/085611 patent/WO2015039555A1/zh active Application Filing
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102397563A (zh) * | 2010-09-16 | 2012-04-04 | 同济大学 | 一种磁共振成像造影剂用石墨烯纳米载体的制备方法 |
US20130079503A1 (en) * | 2011-09-08 | 2013-03-28 | Research & Business Foundation Sungkyunkwan University | Go-gd-dtpa complex, preparation method thereof, and mri contrast agent comprising the same |
CN102657872A (zh) * | 2012-05-05 | 2012-09-12 | 上海师范大学 | 氧化石墨烯/PAMAM/DTPA-Gd/PSCA抗体多功能材料及其制备方法和应用 |
Non-Patent Citations (1)
Title |
---|
SHEN HE ET AL.: "Application of Graphene in Biomedical Fields", JOURNAL OF SOUTHEAST UNIVERSITY (MEDICAL SCIENCE EDITION, vol. 30, no. 1, 28 February 2011 (2011-02-28) * |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2017066268A1 (en) * | 2015-10-12 | 2017-04-20 | Ohio University | Supercapacitor-based biosensors and methods of detecting a biomarker |
CN108042809A (zh) * | 2018-02-01 | 2018-05-18 | 南京林业大学 | 一种抑菌功能化氧化石墨烯的制备方法 |
Also Published As
Publication number | Publication date |
---|---|
CN104436221A (zh) | 2015-03-25 |
CN104436221B (zh) | 2018-03-20 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
WO2015039555A1 (zh) | 基于氧化石墨烯材料的造影剂及其制备方法 | |
Chee et al. | Biocompatible peptide-coated ultrasmall superparamagnetic iron oxide nanoparticles for in vivo contrast-enhanced magnetic resonance imaging | |
Hou et al. | Nanoparticles for multi-modality cancer diagnosis: simple protocol for self-assembly of gold nanoclusters mediated by gadolinium ions | |
Li et al. | Facile one-pot synthesis of Fe3O4@ Au composite nanoparticles for dual-mode MR/CT imaging applications | |
Rammohan et al. | Nanodiamond–gadolinium (III) aggregates for tracking cancer growth in vivo at high field | |
Nam et al. | Tumor targeting chitosan nanoparticles for dual-modality optical/MR cancer imaging | |
Bae et al. | Bioinspired synthesis and characterization of gadolinium-labeled magnetite nanoparticles for dual contrast T 1-and T 2-weighted magnetic resonance imaging | |
Wei et al. | Compact zwitterion-coated iron oxide nanoparticles for biological applications | |
Lee et al. | Prostate cancer-targeted imaging using magnetofluorescent polymeric nanoparticles functionalized with bombesin | |
Du et al. | PSA targeted dual-modality manganese oxide–mesoporous silica nanoparticles for prostate cancer imaging | |
Johnson et al. | Compact micellization: a strategy for ultrahigh T1 magnetic resonance contrast with gadolinium-based nanocrystals | |
Song et al. | Facile synthesis of gold nanospheres modified by positively charged mesoporous silica, loaded with near-infrared fluorescent dye, for in vivo X-ray computed tomography and fluorescence dual mode imaging | |
Li et al. | Multifunctional surface modification of gold-stabilized nanoparticles by bioorthogonal reactions | |
Fan et al. | Design and Biomedical Applications of Poly (amidoamine)‐Dendrimer‐Based Hybrid Nanoarchitectures | |
Chen et al. | Gadolinium-conjugated PLA-PEG nanoparticles as liver targeted molecular MRI contrast agent | |
Otis et al. | Dendrimer antibody conjugate to target and image HER-2 overexpressing cancer cells | |
Jing et al. | Multifunctional nanoflowers for simultaneous multimodal imaging and high-sensitivity chemo-photothermal treatment | |
Hemalatha et al. | Fabrication and characterization of dual acting oleyl chitosan functionalised iron oxide/gold hybrid nanoparticles for MRI and CT imaging | |
US20100209353A1 (en) | Tumor targeting protein conjugate and a method for preparing the same | |
CN104436220B (zh) | 一种壳聚糖磁性纳米微球的制备方法及其用途 | |
US20110064676A1 (en) | Diagnostic and therapeutic nanoparticles | |
CN101979096B (zh) | 负载金和碘元素的树状大分子ct靶向造影剂及其制备 | |
Zhou et al. | Acetylated polyethylenimine-entrapped gold nanoparticles enable negative computed tomography imaging of orthotopic hepatic carcinoma | |
Mehravi et al. | Conjugation of glucosamine with Gd3+-based nanoporous silica using a heterobifunctional ANB-NOS crosslinker for imaging of cancer cells | |
US20140044648A1 (en) | Activatable imaging contrast agents |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
121 | Ep: the epo has been informed by wipo that ep was designated in this application |
Ref document number: 14845406 Country of ref document: EP Kind code of ref document: A1 |
|
NENP | Non-entry into the national phase |
Ref country code: DE |
|
122 | Ep: pct application non-entry in european phase |
Ref document number: 14845406 Country of ref document: EP Kind code of ref document: A1 |