CN112353953A - Broad-spectrum bacteria targeted magnetic resonance imaging contrast agent and preparation method and application thereof - Google Patents
Broad-spectrum bacteria targeted magnetic resonance imaging contrast agent and preparation method and application thereof Download PDFInfo
- Publication number
- CN112353953A CN112353953A CN202011237671.1A CN202011237671A CN112353953A CN 112353953 A CN112353953 A CN 112353953A CN 202011237671 A CN202011237671 A CN 202011237671A CN 112353953 A CN112353953 A CN 112353953A
- Authority
- CN
- China
- Prior art keywords
- magnetic resonance
- resonance imaging
- contrast agent
- broad
- bacteria
- 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.)
- Pending
Links
- 239000002405 nuclear magnetic resonance imaging agent Substances 0.000 title claims abstract description 34
- 241000894006 Bacteria Species 0.000 title claims abstract description 33
- 238000002360 preparation method Methods 0.000 title abstract description 10
- 238000002595 magnetic resonance imaging Methods 0.000 claims abstract description 22
- 239000002872 contrast media Substances 0.000 claims abstract description 19
- 206010028980 Neoplasm Diseases 0.000 claims abstract description 10
- 239000002105 nanoparticle Substances 0.000 claims description 60
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 claims description 26
- 229920002774 Maltodextrin Polymers 0.000 claims description 25
- 239000005913 Maltodextrin Substances 0.000 claims description 25
- 229940035034 maltodextrin Drugs 0.000 claims description 25
- 150000002482 oligosaccharides Chemical class 0.000 claims description 22
- 229920001542 oligosaccharide Polymers 0.000 claims description 21
- 239000013522 chelant Substances 0.000 claims description 14
- UIWYJDYFSGRHKR-UHFFFAOYSA-N gadolinium atom Chemical compound [Gd] UIWYJDYFSGRHKR-UHFFFAOYSA-N 0.000 claims description 14
- 229910052688 Gadolinium Inorganic materials 0.000 claims description 13
- 230000001580 bacterial effect Effects 0.000 claims description 13
- 229910052742 iron Inorganic materials 0.000 claims description 12
- 239000002245 particle Substances 0.000 claims description 12
- 229920000642 polymer Polymers 0.000 claims description 12
- PWHULOQIROXLJO-UHFFFAOYSA-N Manganese Chemical compound [Mn] PWHULOQIROXLJO-UHFFFAOYSA-N 0.000 claims description 11
- 229910052748 manganese Inorganic materials 0.000 claims description 11
- 239000011572 manganese Substances 0.000 claims description 11
- 208000035143 Bacterial infection Diseases 0.000 claims description 10
- 208000022362 bacterial infectious disease Diseases 0.000 claims description 10
- FYGDTMLNYKFZSV-BYLHFPJWSA-N β-1,4-galactotrioside Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@H](CO)O[C@@H](O[C@@H]2[C@@H](O[C@@H](O)[C@H](O)[C@H]2O)CO)[C@H](O)[C@H]1O FYGDTMLNYKFZSV-BYLHFPJWSA-N 0.000 claims description 9
- DBTMGCOVALSLOR-UHFFFAOYSA-N 32-alpha-galactosyl-3-alpha-galactosyl-galactose Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(OC2C(C(CO)OC(O)C2O)O)OC(CO)C1O DBTMGCOVALSLOR-UHFFFAOYSA-N 0.000 claims description 8
- RXVWSYJTUUKTEA-UHFFFAOYSA-N D-maltotriose Natural products OC1C(O)C(OC(C(O)CO)C(O)C(O)C=O)OC(CO)C1OC1C(O)C(O)C(O)C(CO)O1 RXVWSYJTUUKTEA-UHFFFAOYSA-N 0.000 claims description 8
- FYGDTMLNYKFZSV-UHFFFAOYSA-N mannotriose Natural products OC1C(O)C(O)C(CO)OC1OC1C(CO)OC(OC2C(OC(O)C(O)C2O)CO)C(O)C1O FYGDTMLNYKFZSV-UHFFFAOYSA-N 0.000 claims description 8
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 7
- OCIBBXPLUVYKCH-QXVNYKTNSA-N alpha-maltohexaose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)O[C@H](O[C@@H]2[C@H](O[C@H](O[C@@H]3[C@H](O[C@H](O[C@@H]4[C@H](O[C@H](O[C@@H]5[C@H](O[C@H](O)[C@H](O)[C@H]5O)CO)[C@H](O)[C@H]4O)CO)[C@H](O)[C@H]3O)CO)[C@H](O)[C@H]2O)CO)[C@H](O)[C@H]1O OCIBBXPLUVYKCH-QXVNYKTNSA-N 0.000 claims description 6
- DJMVHSOAUQHPSN-UHFFFAOYSA-N malto-hexaose Natural products OC1C(O)C(OC(C(O)CO)C(O)C(O)C=O)OC(CO)C1OC1C(O)C(O)C(OC2C(C(O)C(OC3C(C(O)C(OC4C(C(O)C(O)C(CO)O4)O)C(CO)O3)O)C(CO)O2)O)C(CO)O1 DJMVHSOAUQHPSN-UHFFFAOYSA-N 0.000 claims description 6
- FTNIPWXXIGNQQF-UHFFFAOYSA-N UNPD130147 Natural products OC1C(O)C(O)C(CO)OC1OC1C(CO)OC(OC2C(OC(OC3C(OC(OC4C(OC(O)C(O)C4O)CO)C(O)C3O)CO)C(O)C2O)CO)C(O)C1O FTNIPWXXIGNQQF-UHFFFAOYSA-N 0.000 claims description 5
- BNABBHGYYMZMOA-AHIHXIOASA-N alpha-maltoheptaose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)O[C@H](O[C@@H]2[C@H](O[C@H](O[C@@H]3[C@H](O[C@H](O[C@@H]4[C@H](O[C@H](O[C@@H]5[C@H](O[C@H](O[C@@H]6[C@H](O[C@H](O)[C@H](O)[C@H]6O)CO)[C@H](O)[C@H]5O)CO)[C@H](O)[C@H]4O)CO)[C@H](O)[C@H]3O)CO)[C@H](O)[C@H]2O)CO)[C@H](O)[C@H]1O BNABBHGYYMZMOA-AHIHXIOASA-N 0.000 claims description 5
- FJCUPROCOFFUSR-UHFFFAOYSA-N malto-pentaose Natural products OC1C(O)C(OC(C(O)CO)C(O)C(O)C=O)OC(CO)C1OC1C(O)C(O)C(OC2C(C(O)C(OC3C(C(O)C(O)C(CO)O3)O)C(CO)O2)O)C(CO)O1 FJCUPROCOFFUSR-UHFFFAOYSA-N 0.000 claims description 5
- FJCUPROCOFFUSR-GMMZZHHDSA-N maltopentaose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O[C@H]([C@H](O)CO)[C@H](O)[C@@H](O)C=O)O[C@H](CO)[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](O[C@@H]3[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O3)O)[C@@H](CO)O2)O)[C@@H](CO)O1 FJCUPROCOFFUSR-GMMZZHHDSA-N 0.000 claims description 5
- 238000000034 method Methods 0.000 claims description 5
- LUEWUZLMQUOBSB-UHFFFAOYSA-N UNPD55895 Natural products OC1C(O)C(O)C(CO)OC1OC1C(CO)OC(OC2C(OC(OC3C(OC(O)C(O)C3O)CO)C(O)C2O)CO)C(O)C1O LUEWUZLMQUOBSB-UHFFFAOYSA-N 0.000 claims description 4
- UYQJCPNSAVWAFU-UHFFFAOYSA-N malto-tetraose Natural products OC1C(O)C(OC(C(O)CO)C(O)C(O)C=O)OC(CO)C1OC1C(O)C(O)C(OC2C(C(O)C(O)C(CO)O2)O)C(CO)O1 UYQJCPNSAVWAFU-UHFFFAOYSA-N 0.000 claims description 4
- LUEWUZLMQUOBSB-OUBHKODOSA-N maltotetraose Chemical compound O[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@H](CO)O[C@@H](O[C@@H]2[C@@H](O[C@@H](O[C@@H]3[C@@H](O[C@@H](O)[C@H](O)[C@H]3O)CO)[C@H](O)[C@H]2O)CO)[C@H](O)[C@H]1O LUEWUZLMQUOBSB-OUBHKODOSA-N 0.000 claims description 4
- 238000006243 chemical reaction Methods 0.000 claims description 3
- 150000001875 compounds Chemical class 0.000 claims description 2
- 230000003902 lesion Effects 0.000 claims 2
- 208000015181 infectious disease Diseases 0.000 abstract description 8
- 210000001082 somatic cell Anatomy 0.000 abstract description 5
- 241000192125 Firmicutes Species 0.000 abstract description 4
- 239000012620 biological material Substances 0.000 abstract description 2
- CMIHHWBVHJVIGI-UHFFFAOYSA-N gadolinium(iii) oxide Chemical class [O-2].[O-2].[O-2].[Gd+3].[Gd+3] CMIHHWBVHJVIGI-UHFFFAOYSA-N 0.000 description 63
- 210000004027 cell Anatomy 0.000 description 17
- 239000000243 solution Substances 0.000 description 15
- 238000003384 imaging method Methods 0.000 description 11
- 241000588724 Escherichia coli Species 0.000 description 10
- MTHSVFCYNBDYFN-UHFFFAOYSA-N diethylene glycol Chemical compound OCCOCCO MTHSVFCYNBDYFN-UHFFFAOYSA-N 0.000 description 10
- SZVJSHCCFOBDDC-UHFFFAOYSA-N iron(II,III) oxide Inorganic materials O=[Fe]O[Fe]O[Fe]=O SZVJSHCCFOBDDC-UHFFFAOYSA-N 0.000 description 9
- 238000012986 modification Methods 0.000 description 9
- 230000004048 modification Effects 0.000 description 8
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 7
- 238000002156 mixing Methods 0.000 description 7
- 239000000725 suspension Substances 0.000 description 7
- 238000005406 washing Methods 0.000 description 7
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 6
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 6
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 6
- 238000012512 characterization method Methods 0.000 description 6
- 239000003921 oil Substances 0.000 description 6
- 238000003756 stirring Methods 0.000 description 6
- 239000008367 deionised water Substances 0.000 description 5
- 229910021641 deionized water Inorganic materials 0.000 description 5
- 230000008685 targeting Effects 0.000 description 5
- XDTMQSROBMDMFD-UHFFFAOYSA-N Cyclohexane Chemical compound C1CCCCC1 XDTMQSROBMDMFD-UHFFFAOYSA-N 0.000 description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 4
- BOTDANWDWHJENH-UHFFFAOYSA-N Tetraethyl orthosilicate Chemical compound CCO[Si](OCC)(OCC)OCC BOTDANWDWHJENH-UHFFFAOYSA-N 0.000 description 4
- 238000005119 centrifugation Methods 0.000 description 4
- WOWHHFRSBJGXCM-UHFFFAOYSA-M cetyltrimethylammonium chloride Chemical compound [Cl-].CCCCCCCCCCCCCCCC[N+](C)(C)C WOWHHFRSBJGXCM-UHFFFAOYSA-M 0.000 description 4
- 239000003153 chemical reaction reagent Substances 0.000 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 description 4
- 239000000463 material Substances 0.000 description 4
- GXCLVBGFBYZDAG-UHFFFAOYSA-N N-[2-(1H-indol-3-yl)ethyl]-N-methylprop-2-en-1-amine Chemical compound CN(CCC1=CNC2=C1C=CC=C2)CC=C GXCLVBGFBYZDAG-UHFFFAOYSA-N 0.000 description 3
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 description 3
- GSEJCLTVZPLZKY-UHFFFAOYSA-N Triethanolamine Chemical compound OCCN(CCO)CCO GSEJCLTVZPLZKY-UHFFFAOYSA-N 0.000 description 3
- 230000005284 excitation Effects 0.000 description 3
- 238000000684 flow cytometry Methods 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 230000000242 pagocytic effect Effects 0.000 description 3
- 229910052710 silicon Inorganic materials 0.000 description 3
- 239000010703 silicon Substances 0.000 description 3
- 239000000377 silicon dioxide Substances 0.000 description 3
- 210000004881 tumor cell Anatomy 0.000 description 3
- FPQQSJJWHUJYPU-UHFFFAOYSA-N 3-(dimethylamino)propyliminomethylidene-ethylazanium;chloride Chemical compound Cl.CCN=C=NCCCN(C)C FPQQSJJWHUJYPU-UHFFFAOYSA-N 0.000 description 2
- 208000035473 Communicable disease Diseases 0.000 description 2
- 239000002616 MRI contrast agent Substances 0.000 description 2
- 229930040373 Paraformaldehyde Natural products 0.000 description 2
- 241000191967 Staphylococcus aureus Species 0.000 description 2
- 238000007112 amidation reaction Methods 0.000 description 2
- 239000007864 aqueous solution Substances 0.000 description 2
- 230000005540 biological transmission Effects 0.000 description 2
- -1 carboxyl modified gadolinium oxide Chemical class 0.000 description 2
- 239000006285 cell suspension Substances 0.000 description 2
- 238000004624 confocal microscopy Methods 0.000 description 2
- 238000010276 construction Methods 0.000 description 2
- 210000004292 cytoskeleton Anatomy 0.000 description 2
- 238000003748 differential diagnosis Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 229910001938 gadolinium oxide Inorganic materials 0.000 description 2
- 229940075613 gadolinium oxide Drugs 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 230000004060 metabolic process Effects 0.000 description 2
- 239000011259 mixed solution Substances 0.000 description 2
- 229920002866 paraformaldehyde Polymers 0.000 description 2
- 238000010992 reflux Methods 0.000 description 2
- 230000035945 sensitivity Effects 0.000 description 2
- 238000004627 transmission electron microscopy Methods 0.000 description 2
- GXDMUOPCQNLBCZ-UHFFFAOYSA-N 3-(3-triethoxysilylpropyl)oxolane-2,5-dione Chemical compound CCO[Si](OCC)(OCC)CCCC1CC(=O)OC1=O GXDMUOPCQNLBCZ-UHFFFAOYSA-N 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- NQTADLQHYWFPDB-UHFFFAOYSA-N N-Hydroxysuccinimide Chemical compound ON1C(=O)CCC1=O NQTADLQHYWFPDB-UHFFFAOYSA-N 0.000 description 1
- 241000589517 Pseudomonas aeruginosa Species 0.000 description 1
- 229920002125 Sokalan® Polymers 0.000 description 1
- 102000004142 Trypsin Human genes 0.000 description 1
- 108090000631 Trypsin Proteins 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 230000001464 adherent effect Effects 0.000 description 1
- FYGDTMLNYKFZSV-DZOUCCHMSA-N alpha-D-Glcp-(1->4)-alpha-D-Glcp-(1->4)-D-Glcp Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)O[C@H](O[C@@H]2[C@H](OC(O)[C@H](O)[C@H]2O)CO)[C@H](O)[C@H]1O FYGDTMLNYKFZSV-DZOUCCHMSA-N 0.000 description 1
- 238000005576 amination reaction Methods 0.000 description 1
- 239000003054 catalyst Substances 0.000 description 1
- 239000003093 cationic surfactant Substances 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 239000006143 cell culture medium Substances 0.000 description 1
- 210000003855 cell nucleus Anatomy 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000001218 confocal laser scanning microscopy Methods 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000000502 dialysis Methods 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 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
- 230000009881 electrostatic interaction Effects 0.000 description 1
- 239000003995 emulsifying agent Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 238000002329 infrared spectrum Methods 0.000 description 1
- 208000014674 injury Diseases 0.000 description 1
- 150000002505 iron Chemical class 0.000 description 1
- WPBNNNQJVZRUHP-UHFFFAOYSA-L manganese(2+);methyl n-[[2-(methoxycarbonylcarbamothioylamino)phenyl]carbamothioyl]carbamate;n-[2-(sulfidocarbothioylamino)ethyl]carbamodithioate Chemical compound [Mn+2].[S-]C(=S)NCCNC([S-])=S.COC(=O)NC(=S)NC1=CC=CC=C1NC(=S)NC(=O)OC WPBNNNQJVZRUHP-UHFFFAOYSA-L 0.000 description 1
- 230000002906 microbiologic effect Effects 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 210000004940 nucleus Anatomy 0.000 description 1
- 201000008968 osteosarcoma Diseases 0.000 description 1
- 238000004917 polyol method Methods 0.000 description 1
- 238000002600 positron emission tomography Methods 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 238000012205 qualitative assay Methods 0.000 description 1
- 238000004445 quantitative analysis Methods 0.000 description 1
- 230000005855 radiation Effects 0.000 description 1
- 238000012552 review Methods 0.000 description 1
- 108700038288 rhodamine-phalloidin Proteins 0.000 description 1
- 210000004872 soft tissue Anatomy 0.000 description 1
- 230000003595 spectral effect Effects 0.000 description 1
- 238000003325 tomography Methods 0.000 description 1
- 230000008733 trauma Effects 0.000 description 1
- 239000012588 trypsin Substances 0.000 description 1
Images
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
- A61K49/08—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by the carrier
- A61K49/10—Organic compounds
-
- 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
- A61K49/18—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
- A61K49/1827—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 having a (super)(para)magnetic core, being a solid MRI-active material, e.g. magnetite, or composed of a plurality of MRI-active, organic agents, e.g. Gd-chelates, or nuclei, e.g. Eu3+, encapsulated or entrapped in the core of the coated or functionalised nanoparticle
- A61K49/1833—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 having a (super)(para)magnetic core, being a solid MRI-active material, e.g. magnetite, or composed of a plurality of MRI-active, organic agents, e.g. Gd-chelates, or nuclei, e.g. Eu3+, encapsulated or entrapped in the core of the coated or functionalised nanoparticle having a (super)(para)magnetic core coated or functionalised with a small organic molecule
- A61K49/1845—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 having a (super)(para)magnetic core, being a solid MRI-active material, e.g. magnetite, or composed of a plurality of MRI-active, organic agents, e.g. Gd-chelates, or nuclei, e.g. Eu3+, encapsulated or entrapped in the core of the coated or functionalised nanoparticle having a (super)(para)magnetic core coated or functionalised with a small organic molecule the small organic molecule being a carbohydrate (monosaccharides, discacharides)
-
- 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
- A61K49/1827—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 having a (super)(para)magnetic core, being a solid MRI-active material, e.g. magnetite, or composed of a plurality of MRI-active, organic agents, e.g. Gd-chelates, or nuclei, e.g. Eu3+, encapsulated or entrapped in the core of the coated or functionalised nanoparticle
- A61K49/1851—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 having a (super)(para)magnetic core, being a solid MRI-active material, e.g. magnetite, or composed of a plurality of MRI-active, organic agents, e.g. Gd-chelates, or nuclei, e.g. Eu3+, encapsulated or entrapped in the core of the coated or functionalised nanoparticle having a (super)(para)magnetic core coated or functionalised with an organic macromolecular compound, i.e. oligomeric, polymeric, dendrimeric organic molecule
- A61K49/1863—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 having a (super)(para)magnetic core, being a solid MRI-active material, e.g. magnetite, or composed of a plurality of MRI-active, organic agents, e.g. Gd-chelates, or nuclei, e.g. Eu3+, encapsulated or entrapped in the core of the coated or functionalised nanoparticle having a (super)(para)magnetic core coated or functionalised with an organic macromolecular compound, i.e. oligomeric, polymeric, dendrimeric organic molecule the organic macromolecular compound being a polysaccharide or derivative thereof, e.g. chitosan, chitin, cellulose, pectin, starch
Abstract
The invention discloses a broad-spectrum bacteria targeted magnetic resonance imaging contrast agent, a preparation method and application thereof, belonging to the field of medical biomaterials; the broad-spectrum bacteria targeted magnetic resonance imaging contrast agent is enriched in various infection focuses caused by gram-positive bacteria and gram-negative bacteria infection in a targeted manner, and meanwhile, the nonspecific enrichment of the contrast agent in somatic cells such as tumors can be reduced, so that the broad-spectrum bacteria targeted magnetic resonance imaging contrast agent can be used for magnetic resonance imaging identification of the bacteria infection focuses and the tumor focuses.
Description
Technical Field
The invention belongs to the field of medical biomaterials, and particularly relates to a broad-spectrum bacteria targeted magnetic resonance imaging contrast agent, and a preparation method and application thereof.
Background
Bacterial infection is a common clinical infectious disease and seriously threatens the life health and safety of people. In order to effectively judge whether the focus part is infected by bacteria in time, it is necessary to develop a contrast agent with bacterial targeting. Currently, imaging means for clinical application mainly include Positron Emission Tomography (PET), electron tomography (CT), Magnetic Resonance Imaging (MRI), etc., wherein MRI has the advantages of no trauma, no radiation damage, multi-aspect slice imaging, high soft tissue resolution, etc., and becomes clinically important imaging means (FEMS microbiological Reviews,2015,39: 892-. In order to further improve the imaging sensitivity and resolution of MRI, researchers developed gadolinium-, manganese-, and iron-based MRI contrast agents (angelsite chemistry-International Edition,2020,132(6): 2435-. If the bacterial infection can be diagnosed timely and effectively in the early stage of infection, the generation rate of drug-resistant bacteria can be effectively reduced, and the cure rate of infectious diseases can be improved. Therefore, in order to realize the rapid differential diagnosis of bacterial infection diseases, the construction of the MRI contrast agent with the broad-spectrum bacterial targeting function has very important significance.
Maltodextrin oligosaccharides such as maltotriose, maltopentaose, maltohexaose, maltoheptaose and the like can be used as a glucose source and actively participate in the metabolic process of bacteria through a maltodextrin transport system of the bacteria, so as to target the bacterial infection site in vivo with high specificity and high sensitivity (Nature Materials,2011,10(8): 602-607; Angewandte Chemie-International Edition,2014,53(51): 14096-. In addition, since human somatic cells do not express maltodextrin transporters, they do not actively phagocytose maltodextrin oligosaccharide-based contrast agents. Therefore, the maltodextrin oligosaccharide has good targeting property on various gram-positive bacteria and gram-negative bacteria.
Disclosure of Invention
A first object of the present invention is to solve the above problems in the prior art, and to provide a broad-spectrum bacteria-targeted magnetic resonance imaging contrast agent; the second purpose of the invention is to provide a preparation method of the contrast agent; the third purpose of the invention is to provide the application of the contrast agent in the magnetic resonance imaging identification of bacterial infection focuses and tumor focuses.
The first object of the present invention can be achieved by the following technical solutions: a broad-spectrum bacteria-targeted magnetic resonance imaging contrast agent is characterized by comprising
A contrast agent having a magnetic resonance imaging function;
bacterial target molecule maltodextrin oligosaccharide, which is used for modifying the contrast agent with the magnetic resonance imaging function.
Preferably, the contrast agent with the magnetic resonance imaging function is gadolinium-based polymer chelate or manganese-based polymer chelate or iron-based polymer chelate or gadolinium-based nanoparticle or manganese-based nanoparticle or iron-based nanoparticle.
Preferably, the gadolinium-based polymer chelate or manganese-based polymer chelate or iron-based polymer chelate has a molecular weight of less than 3000.
Preferably, the particle size of the gadolinium-based nanoparticles, manganese-based nanoparticles or iron-based nanoparticles is 1-400 nm.
Preferably, the bacterial target molecule maltodextrin oligosaccharide is one or more of maltotriose, maltotetraose, maltopentaose, maltohexaose, maltoheptaose, a modified product of maltotriose, a modified product of maltotetraose, a modified product of maltopentaose, a modified product of maltohexaose, and a modified product of maltoheptaose.
Preferably, in the broad-spectrum bacteria-targeted magnetic resonance imaging contrast agent, the content of the bacteria target molecule maltodextrin oligosaccharide is 0.01-5 wt% based on the total weight of the compound.
Preferably, the zeta potential of the magnetic resonance imaging contrast agent of the broad-spectrum bacteria-targeted type is from-40 mV to 40 mV.
Preferably, the longitudinal relaxation efficiency r of the broad-spectrum bacteria-targeted magnetic resonance imaging contrast agent1A value of 0.5mM- 1s-1～50mM-1s-1Transverse relaxation efficiency r2A value of 1mM-1s-1～300mM-1s-1。
The second object of the present invention can be achieved by the following technical solutions: the preparation method of the broad-spectrum bacterium targeted magnetic resonance imaging contrast agent is characterized by comprising the following steps of:
step (1): dispersing a contrast agent with a magnetic resonance imaging function in water to obtain a solution;
step (2): adding bacterial target molecules maltodextrin oligosaccharide in the step (1) for reaction to obtain the broad-spectrum bacterial targeted magnetic resonance imaging contrast agent.
The third object of the present invention can be achieved by the following technical solutions: the broad-spectrum bacteria-targeted magnetic resonance imaging contrast agent is applied to the magnetic resonance imaging identification of bacterial infection focuses and tumor focuses.
The working principle of the invention is as follows: the gadolinium-based, manganese-based and iron-based chelate or nanoparticle is used as an excellent magnetic resonance imaging contrast agent, the maltodextrin oligosaccharide is used as target molecules of various bacteria, and the maltodextrin oligosaccharide is modified on a gadolinium-based, iron-based or manganese-based material to construct the magnetic resonance imaging contrast agent with targeting performance on various bacteria; after the contrast agent is injected into a body, maltodextrin oligosaccharide actively participates in the metabolic process of bacteria through a maltodextrin transport system of the bacteria, so that the contrast agent is subjected to targeted enrichment on bacterial infection focuses, and the magnetic resonance imaging signals of the bacterial infection focuses are enhanced; since somatic cells such as tumor cells do not express maltodextrin transporters, the somatic cells do not actively phagocytose maltodextrin oligosaccharide-based contrast agents.
Compared with the prior art, the invention has the following advantages:
the broad-spectrum bacteria targeted magnetic resonance imaging contrast agent is prepared by reacting gadolinium-based, manganese-based and iron-based micromolecule chelate or nanoparticle with maltodextrin oligosaccharide, and the preparation method is simple and easy to implement.
The broad-spectrum bacteria targeted magnetic resonance imaging contrast agent can be enriched in various infection focuses caused by gram-positive bacteria and gram-negative bacteria infection in a targeted manner, and meanwhile, the nonspecific enrichment of the contrast agent in somatic cells such as tumors can be reduced, so that the broad-spectrum bacteria targeted magnetic resonance imaging contrast agent can be used for magnetic resonance imaging identification of the bacteria infection focuses and the tumor focuses.
The construction of the broad-spectrum bacteria targeted magnetic resonance imaging contrast agent is expected to realize the differential diagnosis of the bacterial infection focus and the tumor focus by the magnetic resonance imaging technology.
Drawings
FIG. 1 is a diagram of maltotriose (G3) -modified gadolinium oxide nanoparticles (Gd)2O3-G3);
FIG. 2 shows gadolinium-based nanoparticles (Gd) modified with malto-oligosaccharides2O3@ MSN-Malt) profile of contrast agents;
FIG. 3 is Gd2O3A hydrated particle size characterization plot of @ MSN-Malt;
FIG. 4 is Gd2O3Zeta potential profile of @ MSN-Malt;
FIG. 5 is Gd2O3@ MSN-Malt longitudinal relaxation efficiency (r)1) And transverse relaxation efficiency (r)2) A result graph;
FIG. 6 is Gd2O3T of @ MSN-Malt1-a weighted imaging profile;
FIG. 7 is a confocal microscope qualitative assay of Gd in tumor cells2O3The phagocytic capacity profile of @ MSN-Malt;
FIG. 8 is a flow cytometer for quantitatively detecting Gd in tumor cells2O3Graph of phagocytic potency of @ MSN-Malt;
FIG. 9 is Gd2O3@ MSN-Malt on gram-positive bacteria (Staphylococcus aureus, S.aureus) and gram-negative bacteriaSchematic representation of the targeting of the bacteria (e.coli).
Detailed Description
In order to make the technical solutions of the present invention better understood, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it should be understood that the following embodiments are only for illustrating the present invention and do not limit the present invention.
In the examples, Tecnai F20 from FEI, USA was used as the transmission electron microscope.
In the examples, the hydrated particle size and zeta-potential of the contrast agent were measured using a dynamic light scattering particle size analyzer Zetasizer Nano ZS from malvern instruments ltd, uk.
In the examples, Verios G4 UC from Thermo Scientific, USA is used as the scanning electron microscope.
In the examples, a Philips-Ingenia 1.5T MR imager was used for relaxation performance testing and magnetic resonance imaging testing.
In the examples, the phagocytic capacity of Gd2O3@ MSN-Malt of tumor cells was characterized using a laser confocal microscope (LSM-710) from McMac instruments, USA and a flow cytometer (BD LSRFortessa) from BD USA.
The present invention will be described in detail by further examples. It is also to be understood that the following examples are illustrative of the present invention and are not to be construed as limiting the scope of the invention, and that certain insubstantial modifications and adaptations of the invention by those skilled in the art in light of the foregoing description are intended to be included within the scope of the invention; the specific process parameters and the like of the following examples are also only one example of suitable ranges, i.e., those skilled in the art can select the appropriate ranges through the description herein, and are not limited to the specific values exemplified below.
Example 1
Maltotriose-modified gadolinium oxide nanoparticles (Gd)2O3-G3) preparation and morphology characterization
(1) The carboxyl modified gadolinium oxide nano-particles are prepared by adopting a polyol method, and the specific method is as follows: mixing 0.10mmol polyacrylic acid (PAA) and 0.8mmol Gd (NO)3)3·6H2Adding O into 10mL of diethylene glycol, and stirring for 2 hours at the temperature of 60 ℃; dissolving 5mmol of NaOH in 5mL of diethylene glycol, and slowly adding the solution; adjusting the temperature to 110 ℃, and continuing to react for 12 hours; cooling to room temperature, dialyzing the product, and concentrating to obtain PAA-modified gadolinium oxide nanoparticles (PAA-Gd)2O3)。
(2) Maltotriose amination modification: 0.4mmol of maltotriose was dissolved in 5ml of NH4To the OH solution, 0.4mmol of NH was subsequently added4HCO3Refluxing in an oil bath at 42 ℃ for 36h, concentrating, and freeze-drying to obtain the amino modified maltotriose Named (NH)2-G3)。
(3) Mixing PAA-Gd2O3Dispersing in deionized water, adding 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (EDC) and N-N-hydroxysuccinimide (NHS), and stirring at 0 deg.C for 2 hr; 0.005g of NH were added2G3, stirring overnight, removing unreacted reagent by multiple centrifugations and washes to finally obtain Gd2O3-G3. The morphology of the nanoparticles was characterized by transmission electron microscopy and the results are shown in fig. 1: the particle size of the nanoparticles is about 2-5 nm.
Example 2
Gadolinium oxide loaded mesoporous silica nanoparticles (Gd) modified by malto-oligosaccharide2O3Preparation of @ MSN-Malt)
(1) The method is characterized in that a two-phase method is adopted to prepare Mesoporous Silica (MSN) nanoparticles with a macroporous structure, wherein a cationic surfactant, namely hexadecyl trimethyl ammonium chloride (CTAC), is used as a template, Tetraethoxysilane (TEOS) is used as a silicon source, Triethanolamine (TEA) is used as a catalyst, and cyclohexane is used as an emulsifier. The preparation method comprises the following steps: firstly, 6g of CTAC and 0.18g of TEA are dissolved in 60mL of deionized water and stirred in an oil bath kettle at 60 ℃ for 2 hours to form a uniform solution; then 2mL of TEOS is dissolved in 20mL of cyclohexane to form a cyclohexane solution with TEOS concentration of 10% (v/v%), the cyclohexane solution is dripped into the mixed solution, and the mixture is continuously stirred for 18 hours in an oil bath kettle at the temperature of 60 ℃; collecting the product by centrifugation at 12000rpm for 10 min, and washing with ethanol and deionized water to remove unreacted reagent; finally, the product was dispersed in a solution of methanol in HCl (methanol: HCl ═ 10:1) and refluxed in an oil bath at 80 ℃ for 8 hours to remove the templating agent CTAC, then the product was dispersed in deionized water and stored in a refrigerator at 4 ℃ until use.
(2) Adding 6.0mmol Gd (NO)3)3·6H2Dissolving O in 26mL of diethylene glycol (DEG), and stirring in an oil bath kettle at 100 ℃ until the O is completely dissolved; NaOH powder was dissolved in DEG (10mg/mL), and 34mL of Gd (NO) was slowly added dropwise3)3·6H2In the O solution, the mixed solution is stirred for 1 hour at 140 ℃; the temperature is raised to 175 ℃, and the reaction is continued for 4 hours; removing unreacted reagent by dialysis and concentration to obtain Gd2O3An aqueous solution.
(3) Gd is added2O3Preparing the nano particles into solutions with the concentration of 0.5mg/mL respectively, adding MSN nano particles with the final concentration of 1mg/mL, stirring at room temperature for 6 hours, centrifuging for multiple times, and washing to obtain the gadolinium oxide loaded mesoporous silica nano particles (Gd)2O3@ MSN). Mixing 40mg of Gd2O3@ MSN was dispersed in 20mL DMF, followed by dropwise addition of 0.2mL (3-triethoxysilyl) propylsuccinic anhydride (TESPSA), refluxing in an oil bath at 80 deg.C overnight, washing several times, and centrifuging to obtain Gd2O3@ MSN-COOH; then to Gd2O3@ MSN-COOH for maltooligosaccharide modification: reacting the above Gd2O3@ MSN-COOH was dispersed in 20mL of deionized water, 0.02g of EDC and 0.01g of NHS were added, and stirred at 0 ℃ for 2 hours; 0.01g NH was added2Malt, after stirring overnight, removing unreacted reagents by multiple centrifugation and washing to finally obtain Gd2O3@ MSN-Malt. The morphology of the nanoparticles was characterized by transmission electron microscopy and the results are shown in fig. 2: the nano-particles still have a typical mesoporous morphology, and the particle size is about 80-100 nm. As can be seen from the characterization of the hydrated particle size of the nanoparticles,with the further modification of the surface of the nano-particles, the hydrated particle size tends to gradually increase: as shown in FIG. 3, Gd2O3@ MSN-COOH has a hydrated particle size of about 157.6nm, and further modified aminated maltodextrin oligosaccharide (NH)2Gd obtained after Malt)2O3@ MSN-Malt has a hydrated particle size of about 194.2 nm. Gd is known from the zeta potential characterization of the nanoparticles2O3The zeta potential of @ MSN is about 4.92 mV; as shown in FIG. 4, when Gd2O3After TESPSA is surface-modified with @ MSN, Gd2O3The zeta potential of @ MSN-COOH was about-26.93 mV; finally grafting maltodextrin oligosaccharide to Gd through amidation reaction2O3@ MSN-COOH to obtain Gd2O3@ MSN-Malt, when the zeta potential of the product is about-29.90 mV. In addition, further infrared spectral characterization revealed that Gd2O3The infrared spectrum of @ MSN-Malt has amido bond generated. Thus, by Gd2O3The change of hydrated particle size, zeta-potential and infrared spectrogram of the nanoparticle before and after the modification of @ MSN can be considered that maltodextrin oligosaccharide is successfully modified on Gd through amidation reaction2O3@ MSN nanoparticle surface.
Example 3
(1) Gd is added2O3、Gd2O3@ MSN and Gd2O3@ MSN-Malt formulated into suspensions with Gd concentrations of 0.05, 0.1, 0.2, 0.4, 0.6 and 0.8mM, tested for longitudinal relaxation efficiency (r) by 1.5T MR imager1) Transverse relaxation efficiency (r)2) And T1-weighting the magnetic resonance imaging performance. As shown in fig. 5, the nanoparticles before and after the maltodextrin oligosaccharide modification have a good linear relationship between different gadolinium concentrations and their corresponding relaxation rates: gd (Gd)2O3、Gd2O3@ MSN and Gd2O3R of @ MSN-Malt1Values are 4.306mM each-1s-1、14.817mM-1s-1And 11.482mM-1s-1，r2Values were 5.969mM each-1s-1、15.959mM-1s-1And 20.575mM-1s-1. Wherein, Gd2O3The longitudinal relaxation rate of @ MSN-Malt is slightly lower than that of Gd2O3@MSN，Gd2O3R of @ MSN-Malt2/r1(1.792) higher than Gd2O3R of @ MSN2/r1(1.077)。
(2)Gd2O3、Gd2O3@ MSN and Gd2O3T of @ MSN-Malt1Weighted imaging performance as shown in fig. 6: gd (Gd)2O3@ MSN and Gd2O3The imaging performance of @ MSN-Malt is superior to that of pure Gd2O3A nanoparticle; gd when the Gd concentration is less than 0.4mM2O3T of @ MSN-Malt1The weighted imaging performance is obviously superior to Gd2O3@ MSN, Gd when the Gd concentration is higher than 0.4mM2O3T of @ MSN-Malt1-weighting of imaging contrast and Gd2O3@ MSN is not clearly distinguished. However, for Gd2O3、Gd2O3@ MSN and Gd2O3Quantitative analysis of the imaged image of @ MSN-Malt revealed that Gd2O3The Delta SNR of @ MSN-Malt is obviously higher than that of Gd2O3And Gd2O3@ MSN, indicating Gd2O3@ MSN-Malt has a much superior contrast.
Example 4
Tumor cell pair Gd2O3Phagocytic potency of @ MSN-Malt
(1) For use in confocal microscopy characterization and flow cytometry, Gd is added2O3@ MSN and Gd2O3The @ MSN-Malt nano-particles are respectively connected with fluorescent molecule Fluorescein Isothiocyanate (FITC) to prepare the nano-particles with fluorescence property.
(2) Osteosarcoma cells mnng-HOS were formulated at a concentration of 2.5X 104Per mL suspension, 2mL of the cell suspension was plated on a confocal dish and then placed in a cell incubator (37 ℃ C., 5% CO)2) Incubating for 24 hours; the medium was discarded and 2mL of FITC-Gd at a concentration of 25. mu.g/mL was added to each dish2O3@ MSN and FITC-Gd2O3@ MSN-Malt medium; after 4 hours of incubation, the material suspension was aspirated, PBS was added, blown and washed to remove non-phagocytized nanoparticles, and finally the cells were fixed, stained, and used for confocal microscopy. The LSCM parameters are set as follows: for a cell nucleus (Hoechst) channel, the wavelengths of excitation light are respectively 405nm, and the corresponding wavelengths of emission light are 420-480 nm; for a nanoparticle (FITC) channel, the wavelength of excitation light is 488nm, and the corresponding wavelength of emitted light is 500-540 nm; for the cytoskeleton (rhodamine-phalloidin) channel, the excitation wavelength is 552nm, and the corresponding emission wavelength is 600-660 nm. The results are shown in FIG. 7: as can be seen from the blank control group, mnng-HOS cells tend to aggregate and grow together, and the cytoskeleton and nucleus states are good; FITC-Gd2O3@MSN、FITC-Gd2O3@ MSN-Malt nanoparticles and mnng-HOS were incubated for 4 hours respectively, at Gd2O3A relatively strong green fluorescence signal was observed in the @ MSN group, indicating that mNng-HOS uptake Gd2O3@ MSN nanoparticles are more; after Malt modification, a weaker green fluorescence signal was observed, indicating that mnng-HOS cells uptake Gd2O3@ MSN-Malt nanoparticles are few.
(3) Quantitative detection of Gd in mnng-HOS cell pairs by flow cytometry2O3@ MSN and Gd2O3The specific operation of the uptake condition of the @ MSN-Malt nano-particles is as follows: first, mnng-HOS was formulated to a concentration of 2X 1052mL of the cell suspension was planted in a six-well plate cell culture plate at 37 ℃ with 5% CO2Incubating for 24 hours in a cell incubator to ensure that cells grow adherent to the wall; then FITC-Gd2O3@ MSN and FITC-Gd2O3@ MSN-Malt was dispersed in cell culture medium to a concentration of 25. mu.g/mL, and 2mL of the material suspension was used in place of the stock culture, wherein cells not containing the above nanoparticles were used as a blank control; after 4 hours of co-incubation, the cells were washed three times with PBS to remove nanoparticles not taken up by the cells; digesting and removing walls of the cells by using trypsin and centrifugally washing the cells; finally dispersing the collected cells into paraformaldehyde with the concentration of 4%, fixing for 30 minutes, centrifuging,After washing, redispersed in PBS and assayed by flow cytometry. The results are shown in FIG. 8: the mnng-HOS cell has certain fluorescence intensity under laser irradiation, and when the cell further takes in the FITC modified nanoparticles, the fluorescence intensity of the whole cell is enhanced. The results show that Malt is responsible for Gd2O3Modification of @ MSN reduces Gd in mnng-HOS cells2O3The intake of @ MSN-Malt.
Example 5
Respectively diluting staphylococcus aureus (S.aureus) and escherichia coli (E.coli) bacterial liquid to 105CFU/mL; 0.6mL of the above-mentioned bacterial suspension was mixed with 50. mu.g/mL of Gd2O3@ MSN and Gd2O3Mixing the @ MSN-Malt nanoparticle suspension, and culturing in a constant temperature shaking box (37 ℃, 180rpm) for 2 hours; respectively taking 10 mu L of S.aureus and E.coli bacteria liquid treated by nano particles, dropwise adding the S.aureus and E.coli bacteria liquid to the surface of the silicon wafer, adding 4% paraformaldehyde solution for fixing for 30 minutes, respectively dehydrating the silicon wafer by 30, 50, 70, 80, 90, 95 and 100% ethanol step by step, and finally observing Gd by SEM2O3@ MSN and Gd2O3Enriching effect of @ MSN-Malt nanoparticles on s. The results are shown in FIG. 9: blank is respectively E.coli and S.aureus without nano particles, and E.coli and S.aureus are respectively rod-shaped and spherical, and have better shapes; in aqueous solution, the surface of the bacteria is negatively charged, Gd2O3@ MSN nanoparticles have a positive charge (4.92mV) on their surface, and only a small amount of nanoparticles are adsorbed around e.coli and s.aureus by the weak electrostatic interaction between the bacteria and the nanoparticles; and at Gd2O3In the @ MSN-Malt group, a large number of nanoparticles were adsorbed on the surfaces of e.coli and s.aureus, and it is apparent from an enlarged view that two kinds of bacteria, e.coli and s.aureus, were respectively surrounded by spherical nanoparticles. This indicates that after maltodextrin oligosaccharide modification, a large amount of Gd is present2O3@ MSN-Malt can be targeted enriched on e.
Example 6
(1) Ultra small size Fe to have excellent MR imaging performance3O4Reacting the nanoparticles with maltohexaose (G6) to obtainFe3O4-G6. Mixing Fe3O4G6 was formulated into suspensions with Fe concentrations of 0.1, 0.2, 0.4, 0.6, 0.8, 1.0mM and tested for relaxation efficiency and T using a 1.5T MR imager1-weighting the magnetic resonance imaging performance. The results show that Fe3O4G6 still had excellent T1-weighting the magnetic resonance imaging performance.
(2) Mixing Fe3O4、Fe3O4G6 was mixed with s.aureus, pseudomonas aeruginosa (p.aeruginosa) respectively and shaken for 4 hours, followed by centrifugation and washing to obtain nanoparticle-treated s.aureus and p.aeruginosa. Mixing Fe3O4、Fe3O4The S.aureus, P.aeruginosa treated with-G6 was imaged in a 1.5T MR imager and the results showed Fe3O4G6-treated s.aureus and p.aeruginosa both have enhanced T1-weighting the imaging effect; by observing the nanoparticles treated s.aureus and p.aeruginosa through a biological transmission electron microscope, a large number of G6-modified nanoparticles can be phagocytized by s.aureus and p.aeruginosa, which indicates that G6-modified iron-based nanoparticles have excellent bacterial specificity and can target a variety of different bacteria.
It is to be understood that the described embodiments are merely a few embodiments of the invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Claims (10)
1. A broad-spectrum bacteria-targeted magnetic resonance imaging contrast agent, characterized in that: comprises that
A contrast agent having a magnetic resonance imaging function;
bacterial target molecule maltodextrin oligosaccharide, which is used for modifying the contrast agent with the magnetic resonance imaging function.
2. The broad-spectrum bacteria-targeted magnetic resonance imaging contrast agent according to claim 1, wherein: the contrast agent with the magnetic resonance imaging function is gadolinium-based polymer chelate or manganese-based polymer chelate or iron-based polymer chelate or gadolinium-based nano-particles or manganese-based nano-particles or iron-based nano-particles.
3. The broad-spectrum bacteria-targeted magnetic resonance imaging contrast agent according to claim 2, wherein: the molecular weight of the gadolinium-based polymer chelate or manganese-based polymer chelate or iron-based polymer chelate is less than 3000.
4. The broad-spectrum bacteria-targeted magnetic resonance imaging contrast agent according to claim 2, wherein: the particle size of the gadolinium-based nanoparticles, manganese-based nanoparticles or iron-based nanoparticles is 1-400 nm.
5. The broad-spectrum bacteria-targeted magnetic resonance imaging contrast agent according to claim 1, wherein: the bacterial target molecule maltodextrin oligosaccharide is one or more of maltotriose, maltotetraose, maltopentaose, maltohexaose, maltoheptaose, a modified product of maltotriose, a modified product of maltotetraose, a modified product of maltopentaose, a modified product of maltohexaose and a modified product of maltoheptaose.
6. The broad-spectrum bacteria-targeted magnetic resonance imaging contrast agent according to claim 1, wherein: in the broad-spectrum bacteria targeted magnetic resonance imaging contrast agent, the content of the bacteria target molecule maltodextrin oligosaccharide is 0.01-5 wt% based on the total weight of the compound.
7. The broad-spectrum bacteria-targeted magnetic resonance imaging contrast agent according to claim 1, wherein: the zeta potential of the broad-spectrum bacteria targeted magnetic resonance imaging contrast agent is-40 mV to 40 mV.
8. The broad-spectrum bacteria-targeted magnetic resonance imaging contrast agent according to claim 1, wherein:longitudinal relaxation efficiency r of the broad-spectrum bacteria-targeted magnetic resonance imaging contrast agent1A value of 0.5mM-1s-1～50mM-1s-1Transverse relaxation efficiency r2A value of 1mM-1s-1～300mM-1s-1。
9. A method for preparing a broad spectrum bacteria-targeted magnetic resonance imaging contrast agent as claimed in any one of claims 1 to 8, comprising the steps of:
step (1): dispersing a contrast agent with a magnetic resonance imaging function in water to obtain a solution;
step (2): adding bacterial target molecules maltodextrin oligosaccharide into the solution obtained in the step (1) for reaction to obtain the broad-spectrum bacterial targeted magnetic resonance imaging contrast agent.
10. Use of a broad spectrum bacteria-targeted magnetic resonance imaging contrast agent according to any one of claims 1-8 for magnetic resonance imaging identification of bacterial infection lesions and tumor lesions.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011237671.1A CN112353953A (en) | 2020-11-09 | 2020-11-09 | Broad-spectrum bacteria targeted magnetic resonance imaging contrast agent and preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011237671.1A CN112353953A (en) | 2020-11-09 | 2020-11-09 | Broad-spectrum bacteria targeted magnetic resonance imaging contrast agent and preparation method and application thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN112353953A true CN112353953A (en) | 2021-02-12 |
Family
ID=74510153
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202011237671.1A Pending CN112353953A (en) | 2020-11-09 | 2020-11-09 | Broad-spectrum bacteria targeted magnetic resonance imaging contrast agent and preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN112353953A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114010797A (en) * | 2021-11-26 | 2022-02-08 | 上海交通大学 | Bacteria-targeted nano antibacterial agent and preparation method thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20140219917A1 (en) * | 2011-01-14 | 2014-08-07 | Emory University | Oligosaccharide conjugates for targeting bacteria and uses related thereto |
| CN108066777A (en) * | 2016-11-17 | 2018-05-25 | 中国科学院大连化学物理研究所 | Cancer target nuclear magnetic resonance-fluorescence supermolecule image-forming contrast medium and preparation and application |
| US20200061215A1 (en) * | 2018-08-23 | 2020-02-27 | The Board Of Trustees Of The Leland Stanford Junior University | Maltotriose-based probes for fluorescence and photoacoustic imaging of bacteria |
| CN111450266A (en) * | 2020-03-30 | 2020-07-28 | 中国科学院大学宁波华美医院 | Magnetic resonance imaging contrast agent targeting gram-positive bacteria, preparation and application |
-
2020
- 2020-11-09 CN CN202011237671.1A patent/CN112353953A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20140219917A1 (en) * | 2011-01-14 | 2014-08-07 | Emory University | Oligosaccharide conjugates for targeting bacteria and uses related thereto |
| CN108066777A (en) * | 2016-11-17 | 2018-05-25 | 中国科学院大连化学物理研究所 | Cancer target nuclear magnetic resonance-fluorescence supermolecule image-forming contrast medium and preparation and application |
| US20200061215A1 (en) * | 2018-08-23 | 2020-02-27 | The Board Of Trustees Of The Leland Stanford Junior University | Maltotriose-based probes for fluorescence and photoacoustic imaging of bacteria |
| CN111450266A (en) * | 2020-03-30 | 2020-07-28 | 中国科学院大学宁波华美医院 | Magnetic resonance imaging contrast agent targeting gram-positive bacteria, preparation and application |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114010797A (en) * | 2021-11-26 | 2022-02-08 | 上海交通大学 | Bacteria-targeted nano antibacterial agent and preparation method thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US11497818B2 (en) | Ultrafine nanoparticles comprising a functionalized polyorganosiloxane matrix and including metal complexes; method for obtaining same and uses thereof in medical imaging and/or therapy | |
| CN103143043B (en) | Preparation method of Fe3O4/Au composite nanoparticles | |
| CN103143041B (en) | Preparation method of targeted MRI (magnetic resonance imaging) contrast medium based on folic acid modified iron oxide nanoparticles | |
| EA015718B1 (en) | Superparamagnetic nanoparticles based on iron oxides with modified surface, method of their preparation and application | |
| US20110158901A1 (en) | Chitosan-based nanoparticles and methods for making and using the same | |
| Kačenka et al. | Dual imaging probes for magnetic resonance imaging and fluorescence microscopy based on perovskite manganite nanoparticles | |
| CN104703625A (en) | Mesoporous oxide nanoparticles and methods of making and using same | |
| CN114377149B (en) | Mn-based degradable MOF nano-reactor and preparation method and application thereof | |
| CN112933251B (en) | Preparation method and application of mangano-manganic oxide nano-particles | |
| US9504761B2 (en) | Stabilized chitosan-based nanoparticles and methods for making the same | |
| Kirla et al. | Carbohydrate coated fluorescent mesoporous silica particles for bacterial imaging | |
| CN104844839B (en) | A kind of preparation method of magnetic fluorescence composite nanometer particle | |
| Mehravi et al. | Acute toxicity evaluation of glycosylated Gd 3+-based silica nanoprobe | |
| CN112353953A (en) | Broad-spectrum bacteria targeted magnetic resonance imaging contrast agent and preparation method and application thereof | |
| CN104274842B (en) | A kind of preparation method of the multi-functional trimanganese tetroxide nano granular core magnetic resonance contrast agent of polyethyleneimine | |
| CN112979530B (en) | Nucleic acid detection probe and preparation method thereof | |
| CN111450266A (en) | Magnetic resonance imaging contrast agent targeting gram-positive bacteria, preparation and application | |
| CN107693803B (en) | Preparation method of manganese oxide-loaded hybrid sodium alginate nanogel | |
| CN109771644B (en) | Nano composite material, preparation method and application | |
| CN114479090B (en) | Fluorinated polyethylene glycol-polyethyleneimine and preparation method and application thereof | |
| CN107929756B (en) | porous Prussian blue nano-particles coated with aminated silicon dioxide as well as preparation method and application of porous Prussian blue nano-particles | |
| CN113105641B (en) | Iron-manganese double-doped nano metal organic framework material and preparation method and application thereof | |
| CN111298139B (en) | Nuclear magnetic imaging guidance-based targeted nano-drug for overcoming drug resistance caused by tumor hypoxia and preparation method and application thereof | |
| Sahoo et al. | Fundamental understanding of the size and surface modification effects on r 1, the relaxivity of Prussian blue nanocube@ m-SiO 2: a novel targeted chemo-photodynamic theranostic agent to treat colon cancer | |
| Eddy et al. | Gadolinium-Mesoporous Silica as a Potential Magnetic Resonance Imaging Contrast Agents |
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 | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20210212 |
|
| RJ01 | Rejection of invention patent application after publication |