CN114949261B - Iron gadolinium nano-composite and preparation method and application thereof - Google Patents
Iron gadolinium nano-composite and preparation method and application thereof Download PDFInfo
- Publication number
- CN114949261B CN114949261B CN202210481034.1A CN202210481034A CN114949261B CN 114949261 B CN114949261 B CN 114949261B CN 202210481034 A CN202210481034 A CN 202210481034A CN 114949261 B CN114949261 B CN 114949261B
- Authority
- CN
- China
- Prior art keywords
- iron
- gadolinium
- near infrared
- nano
- infrared imaging
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- ZSOJHTHUCUGDHS-UHFFFAOYSA-N gadolinium iron Chemical compound [Fe].[Gd] ZSOJHTHUCUGDHS-UHFFFAOYSA-N 0.000 title claims abstract description 90
- 239000002114 nanocomposite Substances 0.000 title claims abstract description 64
- 238000002360 preparation method Methods 0.000 title claims abstract description 6
- 239000002105 nanoparticle Substances 0.000 claims abstract description 41
- 238000003384 imaging method Methods 0.000 claims abstract description 36
- 238000003333 near-infrared imaging Methods 0.000 claims abstract description 32
- 229920000642 polymer Polymers 0.000 claims abstract description 22
- 239000000523 sample Substances 0.000 claims abstract description 18
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 9
- 239000012216 imaging agent Substances 0.000 claims description 20
- 239000002245 particle Substances 0.000 claims description 11
- 210000004881 tumor cell Anatomy 0.000 claims description 10
- 238000006243 chemical reaction Methods 0.000 claims description 9
- 239000002253 acid Substances 0.000 claims description 8
- 238000000034 method Methods 0.000 claims description 8
- MOFVSTNWEDAEEK-UHFFFAOYSA-M indocyanine green Chemical group [Na+].[O-]S(=O)(=O)CCCCN1C2=CC=C3C=CC=CC3=C2C(C)(C)C1=CC=CC=CC=CC1=[N+](CCCCS([O-])(=O)=O)C2=CC=C(C=CC=C3)C3=C2C1(C)C MOFVSTNWEDAEEK-UHFFFAOYSA-M 0.000 claims description 7
- 229960004657 indocyanine green Drugs 0.000 claims description 7
- 239000013067 intermediate product Substances 0.000 claims description 7
- 238000002156 mixing Methods 0.000 claims description 6
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 6
- 238000003756 stirring Methods 0.000 claims description 6
- 239000003960 organic solvent Substances 0.000 claims description 5
- 229920001184 polypeptide Polymers 0.000 claims description 5
- 102000004196 processed proteins & peptides Human genes 0.000 claims description 5
- 230000004913 activation Effects 0.000 claims description 3
- 230000002902 bimodal effect Effects 0.000 claims description 3
- 239000000539 dimer Substances 0.000 claims description 2
- 230000009870 specific binding Effects 0.000 claims 1
- 239000000700 radioactive tracer Substances 0.000 abstract description 3
- 229920001427 mPEG Polymers 0.000 description 40
- 206010028980 Neoplasm Diseases 0.000 description 32
- 239000000243 solution Substances 0.000 description 27
- 239000002872 contrast media Substances 0.000 description 20
- 210000004027 cell Anatomy 0.000 description 18
- 206010027476 Metastases Diseases 0.000 description 14
- 208000005718 Stomach Neoplasms Diseases 0.000 description 14
- 206010017758 gastric cancer Diseases 0.000 description 14
- 230000009401 metastasis Effects 0.000 description 14
- 201000011549 stomach cancer Diseases 0.000 description 14
- 238000002595 magnetic resonance imaging Methods 0.000 description 13
- 230000000694 effects Effects 0.000 description 12
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 11
- 210000001519 tissue Anatomy 0.000 description 9
- 230000008685 targeting Effects 0.000 description 8
- 238000010171 animal model Methods 0.000 description 7
- 238000012360 testing method Methods 0.000 description 7
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 6
- 101100033899 Saccharomyces cerevisiae (strain ATCC 204508 / S288c) RGD2 gene Proteins 0.000 description 6
- 238000003745 diagnosis Methods 0.000 description 6
- 239000007864 aqueous solution Substances 0.000 description 5
- 230000004048 modification Effects 0.000 description 5
- 238000012986 modification Methods 0.000 description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 5
- VHYFNPMBLIVWCW-UHFFFAOYSA-N 4-Dimethylaminopyridine Chemical compound CN(C)C1=CC=NC=C1 VHYFNPMBLIVWCW-UHFFFAOYSA-N 0.000 description 4
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 4
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- -1 IR780 Chemical compound 0.000 description 4
- 239000002616 MRI contrast agent Substances 0.000 description 4
- 201000011510 cancer Diseases 0.000 description 4
- 238000001514 detection method Methods 0.000 description 4
- 238000000799 fluorescence microscopy Methods 0.000 description 4
- 238000001727 in vivo Methods 0.000 description 4
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 3
- 238000005481 NMR spectroscopy Methods 0.000 description 3
- 230000002776 aggregation Effects 0.000 description 3
- 238000004220 aggregation Methods 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 3
- 150000001875 compounds Chemical class 0.000 description 3
- 238000002591 computed tomography Methods 0.000 description 3
- 231100000135 cytotoxicity Toxicity 0.000 description 3
- 230000003013 cytotoxicity Effects 0.000 description 3
- 238000010586 diagram Methods 0.000 description 3
- 239000000975 dye Substances 0.000 description 3
- 230000002496 gastric effect Effects 0.000 description 3
- 238000000338 in vitro Methods 0.000 description 3
- 238000011534 incubation Methods 0.000 description 3
- 230000003902 lesion Effects 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 238000003760 magnetic stirring Methods 0.000 description 3
- 206010061289 metastatic neoplasm Diseases 0.000 description 3
- 238000001000 micrograph Methods 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 230000035945 sensitivity Effects 0.000 description 3
- ADFXKUOMJKEIND-UHFFFAOYSA-N 1,3-dicyclohexylurea Chemical compound C1CCCCC1NC(=O)NC1CCCCC1 ADFXKUOMJKEIND-UHFFFAOYSA-N 0.000 description 2
- GOUHYARYYWKXHS-UHFFFAOYSA-N 4-formylbenzoic acid Chemical compound OC(=O)C1=CC=C(C=O)C=C1 GOUHYARYYWKXHS-UHFFFAOYSA-N 0.000 description 2
- QOSSAOTZNIDXMA-UHFFFAOYSA-N Dicylcohexylcarbodiimide Chemical compound C1CCCCC1N=C=NC1CCCCC1 QOSSAOTZNIDXMA-UHFFFAOYSA-N 0.000 description 2
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 description 2
- 229910052688 Gadolinium Inorganic materials 0.000 description 2
- 108010047852 Integrin alphaVbeta3 Proteins 0.000 description 2
- UQSXHKLRYXJYBZ-UHFFFAOYSA-N Iron oxide Chemical compound [Fe]=O UQSXHKLRYXJYBZ-UHFFFAOYSA-N 0.000 description 2
- 229920002125 Sokalan® Polymers 0.000 description 2
- 230000002378 acidificating effect Effects 0.000 description 2
- 229910021529 ammonia Inorganic materials 0.000 description 2
- 230000005540 biological transmission Effects 0.000 description 2
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 2
- 230000008878 coupling Effects 0.000 description 2
- 238000010168 coupling process Methods 0.000 description 2
- 238000005859 coupling reaction Methods 0.000 description 2
- 238000012377 drug delivery Methods 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- LGMLJQFQKXPRGA-VPVMAENOSA-K gadopentetate dimeglumine Chemical compound [Gd+3].CNC[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO.CNC[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO.OC(=O)CN(CC([O-])=O)CCN(CC([O-])=O)CCN(CC(O)=O)CC([O-])=O LGMLJQFQKXPRGA-VPVMAENOSA-K 0.000 description 2
- 229910052742 iron Inorganic materials 0.000 description 2
- 230000036210 malignancy Effects 0.000 description 2
- 230000001394 metastastic effect Effects 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 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 description 2
- 238000001356 surgical procedure Methods 0.000 description 2
- 230000004083 survival effect Effects 0.000 description 2
- 238000012546 transfer Methods 0.000 description 2
- 210000003462 vein Anatomy 0.000 description 2
- NVHPXYIRNJFKTE-HAGHYFMRSA-N 2-[(2s,5r,8s,11s)-8-(4-aminobutyl)-5-benzyl-11-[3-(diaminomethylideneamino)propyl]-3,6,9,12,15-pentaoxo-1,4,7,10,13-pentazacyclopentadec-2-yl]acetic acid Chemical compound N1C(=O)[C@H](CC(O)=O)NC(=O)CNC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CCCCN)NC(=O)[C@H]1CC1=CC=CC=C1 NVHPXYIRNJFKTE-HAGHYFMRSA-N 0.000 description 1
- 229960000549 4-dimethylaminophenol Drugs 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 108010001857 Cell Surface Receptors Proteins 0.000 description 1
- 102400000888 Cholecystokinin-8 Human genes 0.000 description 1
- 101800005151 Cholecystokinin-8 Proteins 0.000 description 1
- 102000001189 Cyclic Peptides Human genes 0.000 description 1
- 108010069514 Cyclic Peptides Proteins 0.000 description 1
- COLNVLDHVKWLRT-MRVPVSSYSA-N D-phenylalanine Chemical compound OC(=O)[C@H](N)CC1=CC=CC=C1 COLNVLDHVKWLRT-MRVPVSSYSA-N 0.000 description 1
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 206010015548 Euthanasia Diseases 0.000 description 1
- 208000005016 Intestinal Neoplasms Diseases 0.000 description 1
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 description 1
- 206010051676 Metastases to peritoneum Diseases 0.000 description 1
- 208000003788 Neoplasm Micrometastasis Diseases 0.000 description 1
- 206010029155 Nephropathy toxic Diseases 0.000 description 1
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 1
- 239000002262 Schiff base Substances 0.000 description 1
- 150000004753 Schiff bases Chemical class 0.000 description 1
- 238000003917 TEM image Methods 0.000 description 1
- VJMAITQRABEEKP-UHFFFAOYSA-N [6-(phenylmethoxymethyl)-1,4-dioxan-2-yl]methyl acetate Chemical group O1C(COC(=O)C)COCC1COCC1=CC=CC=C1 VJMAITQRABEEKP-UHFFFAOYSA-N 0.000 description 1
- 210000000683 abdominal cavity Anatomy 0.000 description 1
- 230000003187 abdominal effect Effects 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 230000017531 blood circulation Effects 0.000 description 1
- 210000004556 brain Anatomy 0.000 description 1
- 230000005587 bubbling Effects 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 229910002091 carbon monoxide Inorganic materials 0.000 description 1
- 230000003833 cell viability Effects 0.000 description 1
- 239000013522 chelant Substances 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000013170 computed tomography imaging Methods 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 230000008021 deposition Effects 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 238000002296 dynamic light scattering Methods 0.000 description 1
- 210000002889 endothelial cell Anatomy 0.000 description 1
- 210000002615 epidermis Anatomy 0.000 description 1
- 230000005284 excitation Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000012091 fetal bovine serum Substances 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
- 238000001917 fluorescence detection Methods 0.000 description 1
- 239000007850 fluorescent dye Substances 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- UIWYJDYFSGRHKR-UHFFFAOYSA-N gadolinium atom Chemical group [Gd] UIWYJDYFSGRHKR-UHFFFAOYSA-N 0.000 description 1
- RJOJUSXNYCILHH-UHFFFAOYSA-N gadolinium(3+) Chemical compound [Gd+3] RJOJUSXNYCILHH-UHFFFAOYSA-N 0.000 description 1
- ZXQYGBMAQZUVMI-GCMPRSNUSA-N gamma-cyhalothrin Chemical compound CC1(C)[C@@H](\C=C(/Cl)C(F)(F)F)[C@H]1C(=O)O[C@H](C#N)C1=CC=CC(OC=2C=CC=CC=2)=C1 ZXQYGBMAQZUVMI-GCMPRSNUSA-N 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- WGCNASOHLSPBMP-UHFFFAOYSA-N hydroxyacetaldehyde Natural products OCC=O WGCNASOHLSPBMP-UHFFFAOYSA-N 0.000 description 1
- 238000002513 implantation Methods 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 201000002313 intestinal cancer Diseases 0.000 description 1
- 238000002357 laparoscopic surgery Methods 0.000 description 1
- 239000003446 ligand Substances 0.000 description 1
- 230000005923 long-lasting effect Effects 0.000 description 1
- 229920002521 macromolecule Polymers 0.000 description 1
- 102000006240 membrane receptors Human genes 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 238000002324 minimally invasive surgery Methods 0.000 description 1
- 239000003068 molecular probe Substances 0.000 description 1
- 239000002086 nanomaterial Substances 0.000 description 1
- 230000007694 nephrotoxicity Effects 0.000 description 1
- 231100000417 nephrotoxicity Toxicity 0.000 description 1
- 230000009871 nonspecific binding Effects 0.000 description 1
- 239000002405 nuclear magnetic resonance imaging agent Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 230000001936 parietal effect Effects 0.000 description 1
- 238000003921 particle size analysis Methods 0.000 description 1
- 210000004303 peritoneum Anatomy 0.000 description 1
- 239000004584 polyacrylic acid Substances 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 238000010882 preoperative diagnosis Methods 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 108020003175 receptors Proteins 0.000 description 1
- 102000005962 receptors Human genes 0.000 description 1
- 238000010992 reflux Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 238000011076 safety test Methods 0.000 description 1
- IZTQOLKUZKXIRV-YRVFCXMDSA-N sincalide Chemical compound C([C@@H](C(=O)N[C@@H](CCSC)C(=O)NCC(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC=1C=CC=CC=1)C(N)=O)NC(=O)[C@@H](N)CC(O)=O)C1=CC=C(OS(O)(=O)=O)C=C1 IZTQOLKUZKXIRV-YRVFCXMDSA-N 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- 210000004872 soft tissue Anatomy 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 238000002626 targeted therapy Methods 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 238000004627 transmission electron microscopy Methods 0.000 description 1
- 229910021642 ultra pure water Inorganic materials 0.000 description 1
- 239000012498 ultrapure water Substances 0.000 description 1
- 238000010200 validation analysis Methods 0.000 description 1
- 210000003556 vascular endothelial cell Anatomy 0.000 description 1
- 238000012795 verification Methods 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
- A61K49/0002—General or multifunctional contrast agents, e.g. chelated agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/001—Preparation for luminescence or biological staining
- A61K49/0013—Luminescence
- A61K49/0017—Fluorescence in vivo
- A61K49/0019—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules
- A61K49/0021—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules the fluorescent group being a small organic molecule
- A61K49/0032—Methine dyes, e.g. cyanine dyes
- A61K49/0034—Indocyanine green, i.e. ICG, cardiogreen
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/001—Preparation for luminescence or biological staining
- A61K49/0013—Luminescence
- A61K49/0017—Fluorescence in vivo
- A61K49/005—Fluorescence in vivo characterised by the carrier molecule carrying the fluorescent agent
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/001—Preparation for luminescence or biological staining
- A61K49/0013—Luminescence
- A61K49/0017—Fluorescence in vivo
- A61K49/005—Fluorescence in vivo characterised by the carrier molecule carrying the fluorescent agent
- A61K49/0054—Macromolecular compounds, i.e. oligomers, polymers, dendrimers
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/001—Preparation for luminescence or biological staining
- A61K49/0013—Luminescence
- A61K49/0017—Fluorescence in vivo
- A61K49/005—Fluorescence in vivo characterised by the carrier molecule carrying the fluorescent agent
- A61K49/0056—Peptides, proteins, polyamino acids
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/001—Preparation for luminescence or biological staining
- A61K49/0063—Preparation for luminescence or biological staining characterised by a special physical or galenical form, e.g. emulsions, microspheres
- A61K49/0069—Preparation for luminescence or biological staining characterised by a special physical or galenical form, e.g. emulsions, microspheres the agent being in a particular physical galenical form
- A61K49/0089—Particulate, powder, adsorbate, bead, sphere
- A61K49/0091—Microparticle, microcapsule, microbubble, microsphere, microbead, i.e. having a size or diameter higher or equal to 1 micrometer
- A61K49/0093—Nanoparticle, nanocapsule, nanobubble, nanosphere, nanobead, i.e. having a size or diameter smaller than 1 micrometer, e.g. polymeric nanoparticle
-
- 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
- A61K49/12—Macromolecular compounds
- A61K49/126—Linear polymers, e.g. dextran, inulin, PEG
-
- 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
- A61K49/14—Peptides, e.g. proteins
-
- 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
- A61K49/1821—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 coated or functionalised microparticles or nanoparticles
- A61K49/1824—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 coated or functionalised microparticles or nanoparticles coated or functionalised nanoparticles
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y10/00—Nanotechnology for information processing, storage or transmission, e.g. quantum computing or single electron logic
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y15/00—Nanotechnology for interacting, sensing or actuating, e.g. quantum dots as markers in protein assays or molecular motors
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y30/00—Nanotechnology for materials or surface science, e.g. nanocomposites
Landscapes
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Nanotechnology (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Epidemiology (AREA)
- Biomedical Technology (AREA)
- Crystallography & Structural Chemistry (AREA)
- Radiology & Medical Imaging (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Medicinal Chemistry (AREA)
- Physics & Mathematics (AREA)
- Theoretical Computer Science (AREA)
- Mathematical Physics (AREA)
- Molecular Biology (AREA)
- Composite Materials (AREA)
- Condensed Matter Physics & Semiconductors (AREA)
- General Physics & Mathematics (AREA)
- Materials Engineering (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Abstract
The invention provides an iron gadolinium nano-composite and a preparation method and application thereof. The iron-gadolinium nano-composite comprises iron-gadolinium nano-particles, a near infrared imaging fluorescent agent, target molecules and pH sensitive polymers; the iron gadolinium nanoparticle is used as a skeleton to load the near infrared imaging fluorescent agent, the surface of the iron gadolinium nanoparticle is coupled with the target molecule, and the pH sensitive polymer is grafted on the outermost layer. The iron gadolinium nanocomposite disclosed by the invention can be used for preparing a multi-mode imaging probe or a nano tracer.
Description
Technical Field
The invention belongs to the field of nano materials, and particularly relates to an iron-gadolinium nano composite and a preparation method and application thereof.
Background
Gastric cancer is the fifth largest malignancy worldwide, and is also the cause of death associated with the third largest cancer worldwide, posing a serious threat to human health. Gastric cancer is the second and third malignancy in global morbidity and mortality, and about 72 tens of thousands die annually. Recent studies have shown that: laparoscopic minimally invasive surgery can be safely applied to the treatment of locally advanced gastric cancer, however, even though normal standard surgical treatment is performed, up to 20% of patients with advanced gastric cancer develop occult recurrence and metastasis of the abdominal cavity represented by peritoneal implantation. Because the gastric cancer peritoneal metastasis is scattered in multiple cases and the focus is hidden and not easy to find, the tracking and treatment of the gastric cancer peritoneal metastasis are extremely difficult in clinic, and the median survival time is less than 6 months. So how to complete, normalize and efficiently finish the identification of the hidden peritoneal metastasis focus of gastric cancer is a clinical problem to be solved urgently in gastric cancer surgery at present.
At present, computer Tomography (CT) is the most common traditional means for noninvasive diagnosis of peritoneal metastasis, but peritoneal metastasis nodules are soft tissues, small in number, low in volume and density, and limited in various aspects of detection instrument sensitivity, imaging depth, resolution and the like, and generally, the sensitivity of the current CT imaging technology for imaging a metastasis with the diameter of less than 5mm is only 11%, and up to 30% of patients with advanced gastric cancer with negative CT diagnosis peritoneal metastasis are confirmed to be positive in subsequent laparoscopy. In contrast, MRI is capable of detecting metastatic lesions smaller than 2mm, meaning that MRI has a potential advantage in showing microscopic peritoneal metastases, and MRI is likely to be a suitable choice for peritoneal metastasis imaging examination means. Currently, there are two main types of clinically applied MRI contrast agents, T1-weighted MRI contrast agents and T2-weighted MRI contrast agents, and T1 contrast agents have wider application range because they generate bright signals. The most widely used MRI contrast agent clinically is gadolinium chelate T1 contrast agent, but due to its certain nephrotoxicity, brain deposition, the united states Food and Drug Administration (FDA) has warned of its clinical use. Meanwhile, the longitudinal magnetic susceptibility (r 1 value) is low and is only about 4mM -1S-1, so that the MRI imaging effect is poor, and the application of MRI is greatly limited. In addition to pre-operative diagnosis, more than 30% of cases of peritoneal metastasis are found by laparoscopic intra-operative exploration, with a relatively accurate diagnosis of micrometastasis nodules that are difficult to diagnose imagewise, but intra-operative identification of metastatic lesions is highly dependent on the experience and subjective judgment of the operator. Near infrared fluorescence imaging technology based on indocyanine green (ICG) dye is widely applied to imaging of tumors at present, but ICG is taken as a small molecule to be easily dispersed in vivo, and the molecule lacks tumor targeting and can not finish focus specific imaging. Therefore, the research and development of an imaging contrast agent with high biological safety and good imaging efficiency capable of realizing the synchronization of preoperative MRI and intraoperative fluorescence is an effective scheme for improving the prior gastric cancer peritoneal metastasis diagnosis difficulty.
In recent years, more and more tumor specific targets are applied to targeting modification of molecular probes in order to increase aggregation of the probes at tumor sites through active targeting effect, wherein RGD polypeptide modification is an effective targeting strategy. Proved by researches, the RGD polypeptide sequence can be specifically combined with integrin alpha v beta 3 receptors which are highly expressed in various tumor cells and neovascular endothelial cells, and has wide application prospect in molecular drug delivery and tumor targeted therapy. In fact, in addition to tumor cells, some normal cells (e.g., vascular endothelial cells, etc.) also express integrin αvβ3 receptors, suggesting that some nanoparticles may also bind non-specifically to normal tissues, resulting in false positive events, reducing diagnostic accuracy.
Disclosure of Invention
The present invention aims to solve at least one of the technical problems in the prior art described above. Therefore, the first aspect of the invention provides an iron-gadolinium nano-composite which can complete nuclear magnetic resonance and near infrared light dual-mode imaging of a focus under the excitation of nuclear magnetic resonance and fluorescence.
The second aspect of the invention provides a preparation method of the iron gadolinium nanocomposite.
In a third aspect, the invention provides a multi-modality imaging probe.
The fourth aspect of the invention provides a nuclear magnetic resonance near infrared dual-mode imaging nano tracer.
According to a first aspect of the present invention, a method for preparing an iron-gadolinium nanocomposite is provided, the iron-gadolinium nanocomposite comprising iron-gadolinium nanoparticles, a near infrared imaging fluorescent agent, a target molecule, and a pH-sensitive polymer; the iron gadolinium nanoparticle is used as a skeleton to load the near infrared imaging fluorescent agent, and the surface of the iron gadolinium nanoparticle is coupled with the target molecule and is coated by the pH sensitive polymer; the iron gadolinium nano-composite is spherical or spheroid, and the average particle diameter is 10 nm-15 nm.
In the invention, a target molecule is wrapped and protected by utilizing a hydrophilic and good-biocompatibility polymer substance (namely, a pH sensitive polymer connected to the surface), and in a tumor micro-acid environment (pH=6.5-6.8), the polymer is disconnected, so that the target molecule connected with a cancer cell surface receptor is exposed, and the nano-composite specificity is released in the tumor micro-environment; therefore, under the normal cell tissue environment, the compound provided by the invention can reduce the nonspecific uptake of normal cells, but in the tumor slightly acidic environment (pH 6.5-6.8), the receptor on the surface of the tumor cells can be specifically combined with the probe, so that the accurate and efficient aggregation on the tumor part is realized, and finally, the aggregation of nano particles on the tumor part is increased while the normal cytotoxicity is reduced.
In some embodiments of the invention, the near infrared imaging phosphor is loaded at 10% to 15% by weight of the iron gadolinium nanoparticle.
In some embodiments of the invention, the near infrared imaging fluorescent agent is selected from at least one of indocyanine green (Indocyanine Green, ICG), IR780, IR808, IR 820.
In some embodiments of the invention, the target molecule is a target molecule that specifically binds to a tumor cell.
In some preferred embodiments of the invention, the target molecule that specifically binds to tumor cells is an RGD polypeptide or a dimer thereof (H-Glu [ cyclo (Arg-Gly-Asp-D-Phe-Lys) ]2Acetate Form).
In some more preferred embodiments of the invention, the RGD polypeptide comprises an RGD linear peptide or an RGD cyclic peptide.
In some more preferred embodiments of the invention, the tumor cell is at least one of a gastric cancer tumor cell or a intestinal cancer tumor cell.
In some more preferred embodiments of the invention, the pH-sensitive macromolecule is an acid-sensitive amphiphilic polymer having a molecular weight in the range of 5kDa to 10kDa.
Further, the acid-sensitive amphiphilic polymer is selected from at least one of mPEG-FBA and mPEG-CHO.
In the invention, acid-sensitive polymer Schiff base is formed on the surface of the nano particle to perform surface camouflage modification, and the modification degrades and exposes target molecules in a tumor part weak acid environment (pH is 6.5-6.8), so that the tumor part is precisely targeted, and the Trojan horse effect is realized.
According to a second aspect of the present invention, there is provided a method for preparing an iron gadolinium nanocomposite, comprising the steps of:
s1: mixing and reacting the iron gadolinium nano particles serving as a carrier with a near infrared imaging agent to obtain iron gadolinium nano particles loaded with the near infrared imaging agent;
S2: under EDC/NHS activation, mixing and reacting the iron gadolinium nanoparticle loaded with the near infrared imaging agent and target molecules in S1 to obtain an iron gadolinium nanocomposite intermediate product;
s3: and (3) dissolving the intermediate product of the iron-gadolinium nano-composite in the step (S2) and a pH sensitive polymer in an organic solvent, stirring for reaction, and purifying to obtain the iron-gadolinium nano-composite in the first aspect.
In some embodiments of the invention, the molar ratio of the iron gadolinium nanoparticle to the near infrared imaging agent of S1 is (12-1): 1.
Further, the molar ratio of the iron gadolinium nanoparticle to the near infrared imaging agent of S1 is (6-2): 1.
In some preferred embodiments of the invention, the molar ratio of the iron gadolinium nanocomposite intermediate to the target molecule of S2 is (4-2): 1.
Further, the molar ratio of the iron gadolinium nanocomposite intermediate product to the target molecule of S2 is (2-1): 1.
According to a third aspect of the present invention, there is provided a multimodal imaging probe comprising an iron gadolinium nanocomposite according to the first aspect or prepared by the method of the second aspect, the multimodal imaging probe having a diameter of 20nm or less.
According to a fourth aspect of the present invention, there is provided a nuclear magnetic, near infrared bimodal imaging nanotracer comprising an iron gadolinium nanocomposite according to the first aspect or a multimodal imaging probe according to the third aspect.
The beneficial effects of the invention are as follows:
(1) The tumor micro-acid environment response characteristic is realized through the effect of coupling pH sensitive molecular modification, so that the targeting function of the contrast material on tumor cells is improved;
(2) The active targeting molecules are added, the dosage of the contrast agent is reduced, the nonspecific binding with normal tissues is reduced, and the toxic and side effects and the use cost are reduced;
(3) Magnetic resonance and near infrared fluorescence bimodal imaging, realizing high-efficiency imaging before and during operation, and reducing the missed diagnosis rate of tumor diagnosis.
Drawings
The invention is further described with reference to the accompanying drawings and examples, in which:
FIG. 1 is a transmission electron microscope image of FeGdNP and FeGdNP-ICG-RGD2-mPEG prepared in example 1 of the present invention; wherein, the image A is FeGdNP electron microscope image, and the image B is FeGdNP-ICG-RGD2-mPEG electron microscope image.
FIG. 2 is a graph of particle size and potential of FeGdNP and FeGdNP-ICG-RGD2-mPEG prepared in example 1 of the invention; wherein graph A is a particle size plot and graph B is a zeta potential plot.
FIG. 3 is a diagram for investigating biosafety of FeGdNP and FeGdNP-ICG-RGD2-mPEG prepared in example 1 of the present invention.
FIG. 4 is an in vitro active targeting discovery chart of FeGdNP, feGdNP-ICG, feGdNP-ICG-RGD2, feGdNP-ICG-RGD2-mPEG prepared in example 1 of the invention.
FIG. 5 is a graph showing near infrared imaging efficiency verification of FeGdNP and FeGdNP-ICG-RGD2-mPEG prepared in example 1 of the present invention in a peritoneal transfer model.
FIG. 6 is a graph comparing imaging characteristics of FeGdNP-ICG-RGD2-mPEG prepared in example 1 of the present invention with commercially available contrast agents; the left image is a graph of imaging characteristics of a commercially available contrast agent, and the right image is a graph of imaging characteristics of FeGdNP-ICG-RGD2-mPEG prepared in example 1.
FIG. 7 is a diagram of an MRI imaging study of FeGdNP-ICG-RGD2-mPEG prepared in example 1 of the present invention with a commercially available contrast in a peritoneal transfer model; wherein Panel A represents the imaging of a commercially available contrast agent and Panel B represents the imaging of FeGdNP-ICG-RGD2-mPEG prepared in example 1.
Detailed Description
The conception and the technical effects produced by the present invention will be clearly and completely described in conjunction with the embodiments below to fully understand the objects, features and effects of the present invention. It is apparent that the described embodiments are only some embodiments of the present invention, but not all embodiments, and that other embodiments obtained by those skilled in the art without inventive effort are within the scope of the present invention based on the embodiments of the present invention.
Example 1
The embodiment prepares the iron gadolinium nano-composite, which comprises the following specific processes:
(1) First, 20mL of a polyacrylic acid (PAA, mw=1800) solution having a concentration of 4.0mg/mL was purged with nitrogen for 60min to remove oxygen from the solution, followed by heating the polymer solution to reflux to 100 ℃. Then, 0.8mL of the iron-containing precursor mixture (0.5M FeCl 3 and 0.25M FeSO 4) was rapidly injected into the heated polymer solution, followed by adding 6mL of ammonia (28 wt%) and magnetically stirring the mixture at 100 ℃ for 30min to obtain very small magnetic iron oxide nanoparticle (ES-MIONs) cores, then 0.4mLGd (NO 3)3 (0.0625M) and 3.0mL of ammonia (28 wt%) were added to the reaction system, and the reaction was continued for 90min under magnetic stirring at 100 ℃ to finally obtain iron gadolinium nanoparticle (FeGdNP) with MRI contrast function.
(2) Taking 5.0mL of the iron gadolinium nanoparticle solution (C Gd =1.5 mM) synthesized in the step (1), adding 500 mu L of the indocyanine green solution serving as a near infrared imaging agent with the concentration of 0.2mg/mL, magnetically stirring at room temperature for reaction for 4 hours, and storing the obtained nanoparticle (FeGdNP-ICG) loaded with the near infrared imaging agent in a refrigerator at the temperature of 4.0 ℃ for later use.
(3) Coupling a ligand RGD2 taking a cell surface molecule as a target molecule to the nanoparticle surface loaded with the near infrared imaging agent in (2) through reaction of-COOH and-NH 2 under the condition of EDC/NHS activating carboxyl and amino: mu.LEDC and 68. Mu.L NHS (10 mg/mL) were added to 5.6mL of near infrared imaging agent loaded nanoparticles with uninterrupted magnetic stirring, followed by 100. Mu.L (1.0 mg/mL) of RGD2 (Glu- { Cyclo [ Arg-Gly-Asp- (D-Phe) -Lys ] } 2) liquid. Then, after a magnetic stirring reaction at room temperature for 4 hours, the iron gadolinium nanoparticle (FeGdNP-ICG-RGD 2) liquid with the RGD2 coupled on the surface loaded with the near infrared imaging agent is obtained.
(4) Methoxypolyethylene glycol (mPEG, mw=5000) was dissolved in methylene chloride at a concentration of 50mg/mL, and to a 32mL mPEG solution (1.6 g,0.8 mmol), 4-formylbenzoic acid (FBA, 0.36g,2.4 mmol), dimethylformamide (DMF, 4 mL), dicyclohexylcarbodiimide (DCC, 0.496g,2.4 mmol) and 4-dimethylaminopyridine (DMAP, 146mg,1.2 mmol) were added. Stirred at room temperature for 24h, filtered three times and the precipitated product Dicyclohexylurea (DCU) was removed. The organic solvent methylene chloride in the filtrate was then evaporated with nitrogen bubbling, and then the nitrogen-bubbled product was dispersed in 10mL of ultrapure water. The resulting mPEG-FBA was collected by freeze-drying. The resulting mPEG-FBA white powder was dissolved in DMF and stored in a refrigerator at 4 ℃ for later use.
(5) 200 Mu L (4) of the mPEG-FBA (0.4 mg/mL dissolved in DMF) is added into 5.8mL (3) of iron gadolinium nanoparticle solution with RGD2 coupled on the surface loaded with near infrared imaging agent, the obtained product solution is dialyzed with pure water for 48 hours after magnetically stirring reaction at room temperature for 16 hours, the pure water is replaced every 6 hours so as to remove the organic solvent DMF in the liquid, and then the purified nano solution is collected to obtain the iron gadolinium nanocomposite FeGdNP-ICG-RGD2-mPEG.
Test example 1 Transmission Electron microscopy
The iron gadolinium nanocomposite FeGdNP-ICG-RGD2-mPEG solution prepared in example 1 and the iron gadolinium nanoparticle (FeGdNP) solution prepared in example 1 were respectively diluted and then respectively added dropwise onto an ultrathin carbon film, after the solution was naturally evaporated at normal temperature, the sample was sent for inspection, and the original particle size and shape of the nanoparticles were respectively observed by using TEM (transverse electric) as shown in FIGS. 1A and 1B.
As can be seen from the transmission electron microscope results of FIGS. 1A and 1B, feGdNP and FeGdNP-ICG-RGD2-mPEG are spherical nanoparticles, according to TEM image analysis, the average particle sizes of FeGdNP and FeGdNP-ICG-RGD2-mPEG are 5.0nm and 10.0nm respectively, feGdNP and FeGdNP-ICG-RGD2-mPEG are successfully synthesized, and the smaller particles are more easy to enter the tumor part through blood circulation.
Test example 2 particle size and potentiometric analysis
A small amount of the iron gadolinium nanocomposite FeGdNP-ICG-RGD2-mPEG solution prepared in example 1 and the iron gadolinium nanoparticle (FeGdNP) solution prepared in example 1 were taken, respectively, and deionized water was used to prepare a solution of 1:500, and then detecting and analyzing the particle size and the potential condition respectively by using a Markov dynamic light scattering instrument, see fig. 2A and 2B.
As can be seen from the hydrated particle size analysis of FIG. 2A, the particle sizes of FeGdNP and FeGdNP-ICG-RGD2-mPEG are respectively 17.50+ -0.04 nm and 35.01+ -0.05 nm, the hydrated particle size ratio FeGdNP of FeGdNP-ICG-RGD2-mPEG is increased, and the success of the synthesis of the nanocomposite is laterally verified. From the zeta potential of FIG. 2B, it can be seen that the potentials of FeGdNP and FeGdNP-ICG-RGD2-mPEG are-26.76 mV and-15.50 mV, respectively, each negatively charged.
Test example 3 in vitro safety test of nanocomposites
The iron gadolinium nanocomposite FeGdNP-ICG-RGD2-mPEG solution prepared in example 1 and the iron gadolinium nanoparticle (FeGdNP) solution prepared in (1) in example 1 were dispersed in pure water solutions, respectively prepared into FeGdNP-ICG-RGD2-mPEG nanocomposite solutions and FeGdNP solutions with different concentrations (the concentration gradient is set based on the iron ion concentration, the concentrations of each group are respectively 0. Mu.M, 30. Mu.M, 60. Mu.M, 120. Mu.M, 240. Mu.M and 480. Mu.M), and after incubation with normal gastric mucosal cells for 24 hours, the old culture solution was discarded, a complete culture solution containing 10% CCK8 cytotoxicity detection reagent was added, incubation was performed for 4 hours at 37℃under 5% CO 2, and finally absorbance of each culture well at a wavelength of 450nm was detected by an enzyme-labeled instrument, and cell viability was calculated and analyzed as shown in FIG. 3.
From the analysis in fig. 3, feGdNP and FeGdNP-ICG-RGD2-mPEG were found to have cell survival rates of 90% or more after incubation with normal gastric mucosal cells for 24 hours, respectively, which indicates to some extent that both have no significant cytotoxicity to normal gastric mucosal cells, and have potential as good contrast agents.
Test example 4 iron gadolinium nanocomposite FeGdNP-ICG-RGD2-mPEG targeting validation
The iron gadolinium nanocomposite FeGdNP-ICG-RGD2-mPEG prepared in example 1 was selected as the experimental group, and coupled with the biofluorescence dye rhodamine (R6G) as a fluorescent tracer label. The three groups mainly comprise a pure culture medium group (Control), a pure iron gadolinium nanocomposite group (FeGdNP prepared in the example 1) (i.e. FeGdNP-ICG prepared in the example 1) (2), a near infrared imaging agent-loaded group (FeGdNP-ICG prepared in the example 1) (i.e. iron gadolinium nanoparticle FeGdNP-ICG-RGD2 prepared in the example 1 (3) and loaded with the near infrared imaging agent and with RGD2 on the surface, and a PBS-treated iron gadolinium nanocomposite group (FeGdNP-ICG-RGD 2-mPEG pH=6.5) with pH of 6.5) and a PBS-treated iron gadolinium nanocomposite group (FeGdNP-ICG-RGD 2-mPEG pH=7.4) with pH of 7.4. Gastric cancer cells were inoculated into 6-well plates and cultured in DMEM containing 10% fetal bovine serum at 37 ℃ in a 5% co 2 incubator for 24 hours until the cells were completely attached, and the above 6 groups were incubated with gastric cancer cells for 4 hours, respectively. The cells were collected for cell flow detection of fluorescence intensity at a specific wavelength, each group of nanoparticles was labeled with the biofluorescence dye rhodamine (R6G) in advance, the cells of each treatment group were fixed and stained after the same treatment time, and the red fluorescence intensity of the cells of each group at a wavelength of 538nm was observed under a confocal microscope and compared, and the results of the cell fluorescence detection experiment are shown in fig. 4.
As can be seen from FIG. 4, the red fluorescence intensity in the cells of the coupled active target molecule RGD2 treated groups (R6G-FeGdNP-ICG-RGD 2 and R6G-FeGdNP-ICG-RGD 2-mPEG) was significantly higher than that of the non-active target molecule treated groups (R6G-FeGdNP and R6G-FeGdNP-ICG), while the red fluorescence intensity of the nanocomposite treated groups after the acidic condition was significantly higher than that of the normal condition treated group, which was found in the PBS pretreated groups (R6G-FeGdNP-ICG-RGD 2-mPEG) with pH=6.5 and 7.4. In conclusion, the nanocomposite coupled with the active target molecules and the polymer protective outer layer can enter gastric cancer cells in a more targeted manner, and has potential application value in improving drug delivery and tumor imaging efficiency.
Test example 5 in vivo fluorescence imaging Effect of iron gadolinium nanocomposite FeGdNP-ICG-RGD2-mPEG
The experimental animal is prepared for peritoneal tumor bearing, and when the experimental animal peritoneal can touch tumor bodies after 14 days of tumor bearing, the tail vein is injected with the nano probe and the vital signs of the experimental animal are observed. After the probe is injected for 12 hours, euthanasia treatment is carried out on the experimental animal, the peritoneal structure of the experimental animal is fully exposed, the tissue fluorescence imaging effect of the peritoneal tumor body is observed under the irradiation of a near infrared fluorescence imager, and the FLI image acquisition is carried out by adopting a near infrared imaging system as shown in fig. 5.
As can be seen from the imaging fluorescence diagram of FIG. 5, after the near infrared fluorescent probe is injected, the abdominal epidermis of the experimental animal is removed, the peritoneal tissue structure is removed, and besides the fluorescent signal exists in a large piece of adhered tumor tissue, the fluorescence at the peritoneal metastasis site (the circle out site) which is about 3mm beside the tumor tissue is also obvious, so that the peritoneal metastasis site is obviously compared with surrounding normal peritoneal tissue; it is believed that the metastatic tumor tissue at the peritoneum of the mouse parietal layer can be excised precisely under the guidance of the near infrared fluorescence imaging system. In conclusion, the situation shows that the nano-composite has good detection sensitivity and accuracy for distinguishing peritoneal metastasis lesions.
Test example 6 comparison of in vitro magnetic resonance relaxation Rate of iron gadolinium nanocomposite FeGdNP-ICG-RGD2-mPEG with commercial contrast Magnevist solution
And (3) setting iron gadolinium nano-composite FeGdNP-ICG-RGD2-mPEG solution with different concentration gradients and commercial contrast agent aqueous solution, scanning T1 images of the iron gadolinium nano-composite FeGdNP-ICG-RGD2-mPEG solution and commercial contrast agent aqueous solution under a 7.0T magnetic field, calculating and comparing the obtained data to obtain longitudinal relaxation rates of the iron gadolinium nano-composite FeGdNP-ICG-RGD2-mPEG solution and the commercial contrast agent aqueous solution, and predicting the magnetic resonance imaging effect of the iron gadolinium nano-composite. Comparison shows that FeGdNP-ICG-RGD2-mPEG prepared in example 1 has the property that signal intensity increases with increasing concentration. The correlation graphs with 1/T1 for each of the iron gadolinium nanocomposite FeGdNP-ICG-RGD2-mPEG and commercial contrast are shown in FIG. 6 by different concentrations.
The T1 relaxation rates of the two can be obtained through calculation, the r1 value of the commercial contrast agent is found to be 4.21mM -1S-1, the r1 value of the nano imaging probe can reach 13.17mM -1S-1, and the r1 value of the nano imaging probe can reach more than 3 times of the common commercial contrast agent, compared with the conventional commercial contrast agent, the nano imaging probe can ensure better magnetic resonance imaging efficiency under the condition of reducing the concentration of metal gadolinium ions in the contrast agent, and the prepared novel nano imaging probe has good water dispersibility, has higher r1 value than the conventional commercial contrast agent, and can be used for high-contrast tumor T1 weighted magnetic resonance imaging.
Test example 7 comparison of in vivo tumor imaging Effect of iron gadolinium nanocomposite FeGdNP-ICG-RGD2-mPEG with commercial contrast Magnevist
After the experimental animal peritoneal model was successfully constructed, iron gadolinium nanocomposite FeGdNP-ICG-RGD2-mPEG and a commercial aqueous solution of a contrast agent with the same gadolinium ion concentration were set, and in-vivo magnetic resonance images of the iron gadolinium nanocomposite and the commercial aqueous solution were scanned under a 3.0T magnetic field after 12h of tail vein injection, and the result is shown in FIG. 7.
Comparing magnetic resonance signals of the commercial contrast agent and the nano probe in the peritoneal tumor focus, the iron gadolinium nano compound FeGdNP-ICG-RGD2-mPEG treatment group is found to be obviously enhanced in the MRI signal of the tumor area compared with the commercial contrast agent group (the magnetic resonance signal intensity of the synthesized nano imaging probe is brighter in comparison with the magnetic resonance signal intensity of the lower right corner circled part in the figure), which indicates that the iron gadolinium nano compound prepared in the embodiment 1 can realize the contrast enhancement effect of the tumor area. And at the time point of 12 hours, the iron-gadolinium nano-composite can still well gather at the tumor part, so that the iron-gadolinium nano-composite has good magnetic resonance imaging efficiency and simultaneously has the properties of slow release, long accumulation and long-lasting imaging, and is expected to become a good magnetic resonance imaging contrast agent.
While the embodiments of the present invention have been described in detail, the present invention is not limited to the above embodiments, and various changes can be made without departing from the spirit of the present invention within the knowledge of those skilled in the art. Furthermore, embodiments of the invention and features of the embodiments may be combined with each other without conflict.
Claims (7)
1. The iron-gadolinium nano-composite is characterized by comprising iron-gadolinium nano-particles, a near infrared imaging fluorescent agent, target molecules and pH sensitive polymers; the iron gadolinium nanoparticle is used as a skeleton to load the near infrared imaging fluorescent agent, and the surface of the iron gadolinium nanoparticle is coupled with the target molecule and is coated by the pH sensitive polymer; the iron gadolinium nano-composite is spherical or spheroidic, and the average particle diameter is 10 nm-15 nm; the pH sensitive polymer is an acid sensitive amphiphilic polymer; the load of the near infrared imaging fluorescent agent is 10% -15% of the weight of the iron gadolinium nano particles; the molecular weight of the amphiphilic polymer ranges from 5 kDa to 10 kDa; the near infrared imaging fluorescent agent is indocyanine green; the target molecule is RGD polypeptide or dimer thereof, and the acid-sensitive amphiphilic polymer is mPEG-FBA or mPEG-CHO;
The preparation method of the iron gadolinium nano-composite comprises the following steps:
s1: mixing and reacting the iron gadolinium nano particles serving as a carrier with a near infrared imaging agent to obtain iron gadolinium nano particles loaded with the near infrared imaging agent;
S2: under EDC/NHS activation, mixing and reacting the iron gadolinium nanoparticle loaded with the near infrared imaging agent and target molecules in S1 to obtain an iron gadolinium nanocomposite intermediate product;
S3: and (3) dissolving the intermediate product of the iron-gadolinium nano-composite in the step (S2) and a pH sensitive polymer in an organic solvent, stirring for reaction, and purifying to obtain the iron-gadolinium nano-composite.
2. The iron gadolinium nanocomposite according to claim 1, wherein the target molecule is a target molecule having specific binding to tumor cells.
3. A method for preparing the iron gadolinium nanocomposite according to any one of claims 1 to 2, comprising the steps of:
s1: mixing and reacting the iron gadolinium nano particles serving as a carrier with a near infrared imaging agent to obtain iron gadolinium nano particles loaded with the near infrared imaging agent;
S2: under EDC/NHS activation, mixing and reacting the iron gadolinium nanoparticle loaded with the near infrared imaging agent and target molecules in S1 to obtain an iron gadolinium nanocomposite intermediate product;
S3: and (3) dissolving the intermediate product of the iron-gadolinium nano-composite in the step (S2) and a pH sensitive polymer in an organic solvent, stirring for reaction, and purifying to obtain the iron-gadolinium nano-composite.
4. The method for preparing an iron gadolinium nanocomposite according to claim 3, wherein the molar ratio of the iron gadolinium nanoparticle to the near infrared imaging agent is (12-1): 1.
5. A method of preparing an iron gadolinium nanocomposite according to claim 3, wherein the molar ratio of S2 of the iron gadolinium nanocomposite intermediate to the target molecule is (4~2): 1.
6. A multi-modality imaging probe, characterized in that it comprises an iron gadolinium nanocomposite according to any one of claims 1 to 2 or prepared by the method according to any one of claims 3 to 5, the diameter of which is less than or equal to 20 nm.
7. A nuclear magnetic, near infrared bimodal imaging nanotracer, characterized in that it comprises an iron gadolinium nanocomposite according to any one of claims 1-2 or a multimodal imaging probe according to claim 6.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210481034.1A CN114949261B (en) | 2022-05-05 | 2022-05-05 | Iron gadolinium nano-composite and preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210481034.1A CN114949261B (en) | 2022-05-05 | 2022-05-05 | Iron gadolinium nano-composite and preparation method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114949261A CN114949261A (en) | 2022-08-30 |
| CN114949261B true CN114949261B (en) | 2024-06-04 |
Family
ID=82981706
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210481034.1A Active CN114949261B (en) | 2022-05-05 | 2022-05-05 | Iron gadolinium nano-composite and preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114949261B (en) |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2009061456A2 (en) * | 2007-11-07 | 2009-05-14 | University Of Houston | Ultrasmall superparamagnetic iron oxide nanoparticles and uses thereof |
| WO2011112597A1 (en) * | 2010-03-08 | 2011-09-15 | Northeastern University | Magnetic nanoplatforms for theranostic and multi-modal imaging applications |
| CN102743768A (en) * | 2012-07-03 | 2012-10-24 | 中国科学院宁波材料技术与工程研究所 | Stealth contrast-enhancing material for early diagnosis of tumors and preparation method thereof |
| CN102828110A (en) * | 2012-09-19 | 2012-12-19 | 南京信息工程大学 | Low-cobalt nickel nanocrystalline iron-base magnetically soft alloy material and preparation method thereof |
| CN106390143A (en) * | 2015-07-17 | 2017-02-15 | 中国科学院大连化学物理研究所 | Tumor-targeted nuclear magnetic resonance/fluorescence dual-mode imaging contrast agent, preparation and applications thereof |
| CN108392642A (en) * | 2018-02-23 | 2018-08-14 | 莎穆(上海)生物科技有限公司 | A kind of nano-particle magnetic resonance imaging contrast and its preparation method and application containing gadolinium oxide |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2009045579A2 (en) * | 2007-06-14 | 2009-04-09 | The Regents Of The University Of California | Multimodal imaging probes for in vivo targeted and non-targeted imaging and therapeutics |
-
2022
- 2022-05-05 CN CN202210481034.1A patent/CN114949261B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2009061456A2 (en) * | 2007-11-07 | 2009-05-14 | University Of Houston | Ultrasmall superparamagnetic iron oxide nanoparticles and uses thereof |
| WO2011112597A1 (en) * | 2010-03-08 | 2011-09-15 | Northeastern University | Magnetic nanoplatforms for theranostic and multi-modal imaging applications |
| CN102743768A (en) * | 2012-07-03 | 2012-10-24 | 中国科学院宁波材料技术与工程研究所 | Stealth contrast-enhancing material for early diagnosis of tumors and preparation method thereof |
| CN102828110A (en) * | 2012-09-19 | 2012-12-19 | 南京信息工程大学 | Low-cobalt nickel nanocrystalline iron-base magnetically soft alloy material and preparation method thereof |
| CN106390143A (en) * | 2015-07-17 | 2017-02-15 | 中国科学院大连化学物理研究所 | Tumor-targeted nuclear magnetic resonance/fluorescence dual-mode imaging contrast agent, preparation and applications thereof |
| CN108392642A (en) * | 2018-02-23 | 2018-08-14 | 莎穆(上海)生物科技有限公司 | A kind of nano-particle magnetic resonance imaging contrast and its preparation method and application containing gadolinium oxide |
Non-Patent Citations (5)
| Title |
|---|
| Dotted Core-Shell Nanoparticles for T1-Weighted MRI of Tumors;Zheyu Shen，等;《Adv Mater》;第11卷;摘要和正文第3-12、20页 * |
| Indocyanine Green Loaded Magnetic Carbon Nanoparticles for Near Infrared Fluorescence/Magnetic Resonance Dual-Modal Imaging and Photothermal Therapy of Tumor;Saijie Song，等;《ACS Appl Mater Interfaces》;第9卷(第11期);摘要和正文第3-29页 * |
| Multifunctional Theranostic Nanoparticles Based on Exceedingly Small Magnetic Iron Oxide Nanoparticles for T1-Weighted Magnetic Resonance Imaging and Chemotherapy;Zheyu Shen，等;《ACS Nano》;第11卷;摘要和第10992−11001页 * |
| Targeted Magnetic Resonance Imaging/Near-Infrared Dual-Modal Imaging and Ferroptosis/Starvation Therapy of Gastric Cancer with Peritoneal Metastasis;Weihong Guo，等;《Adv. Funct. Mater.》;20230731;第33卷(第27期);2213921 * |
| 铁氧体磁性纳米微粒;余跃东;贵州教育学院学报;20040428(02);全文 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114949261A (en) | 2022-08-30 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Fang et al. | Multifunctional magnetic nanoparticles for medical imaging applications | |
| Hou et al. | Nanodiamond–Manganese dual mode MRI contrast agents for enhanced liver tumor detection | |
| US8916134B2 (en) | Metal nanocomposite, preparation method and use thereof | |
| US20110021745A1 (en) | Ultra-small chitosan nanoparticles useful as bioimaging agents and methods of making same | |
| Hao et al. | Stem cell-mediated delivery of nanogels loaded with ultrasmall iron oxide nanoparticles for enhanced tumor MR imaging | |
| JP2010516760A (en) | Magnetic resonance imaging T1 contrast agent containing manganese oxide nanoparticles | |
| Sun et al. | Controlled nano–bio interface of functional nanoprobes for in vivo monitoring enzyme activity in tumors | |
| Tian et al. | Glypican-3 (GPC3) targeted Fe 3 O 4 core/Au shell nanocomplex for fluorescence/MRI/photoacoustic imaging-guided tumor photothermal therapy | |
| Zhao et al. | Biosynthetic molecular imaging probe for tumor-targeted dual-modal fluorescence/magnetic resonance imaging | |
| Yu et al. | Accurate detection and delineation boundary of renal cell carcinoma based on dual-targeted magnetic-fluorescent carbon dots | |
| CN109395101A (en) | Target the preparation method of the mr contrast agent of blood-brain barrier and glioma | |
| CN106880846B (en) | Tumor-targeted multifunctional nano drug delivery system, preparation method and application | |
| Cui et al. | Gastrin-releasing peptide receptor-targeted gadolinium oxide-based multifunctional nanoparticles for dual magnetic resonance/fluorescent molecular imaging of prostate cancer | |
| CN104815341A (en) | Targeted polymer micelle magnetic nanoparticle, and preparation method and application thereof | |
| Wang et al. | Multifunctional nanoprobe for multi-mode imaging and diagnosis of metastatic prostate cancer | |
| CN109481700A (en) | A kind of molecular probe and preparation method thereof for hepatocarcinoma early diagnosis | |
| CN106310297B (en) | Multifunctional macromolecule prodrug nanoscale medicine delivery system and preparation method and purposes | |
| CN114949261B (en) | Iron gadolinium nano-composite and preparation method and application thereof | |
| KR101042399B1 (en) | Multifunctional iron oxide nanoparticles and a diagnostic agent using the same | |
| Pournoori et al. | Magnetic resonance imaging of tumor‐infiltrating lymphocytes by anti‐CD3‐conjugated iron oxide nanoparticles | |
| Rezayan et al. | A modified PEG-Fe3O4 magnetic nanoparticles conjugated with D (+) glucosamine (DG): MRI contrast agent | |
| CN112870387B (en) | Magnetic nano-drug carrier and preparation method and application thereof | |
| CN109125743B (en) | Targeting nano magnetic resonance contrast agent for glioblastoma multiforme and preparation and application thereof | |
| KR101386715B1 (en) | Advanced biocompatible, non-polymeric surface modified, iron oxide nanoparticles with application of optimal amount of gluconic acid and a composition for diagnosing and treating cancer comprising the same | |
| CN111840324B (en) | Au DENPs-macrophage complex applied to osteosarcoma cell imaging or treatment |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |