CN116903771A - Tumor targeting water-soluble conjugated polymer contrast agent, preparation method and application - Google Patents
Tumor targeting water-soluble conjugated polymer contrast agent, preparation method and application Download PDFInfo
- Publication number
- CN116903771A CN116903771A CN202310734976.0A CN202310734976A CN116903771A CN 116903771 A CN116903771 A CN 116903771A CN 202310734976 A CN202310734976 A CN 202310734976A CN 116903771 A CN116903771 A CN 116903771A
- Authority
- CN
- China
- Prior art keywords
- polymer
- contrast agent
- tumor
- reaction
- solution
- 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
- 229920000547 conjugated polymer Polymers 0.000 title claims abstract description 47
- 206010028980 Neoplasm Diseases 0.000 title claims abstract description 46
- 239000002872 contrast media Substances 0.000 title claims abstract description 44
- 230000008685 targeting Effects 0.000 title claims abstract description 25
- 238000002360 preparation method Methods 0.000 title abstract description 12
- 229920000642 polymer Polymers 0.000 claims abstract description 140
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 30
- 239000000126 substance Substances 0.000 claims abstract description 18
- 150000001413 amino acids Chemical group 0.000 claims abstract description 17
- 238000000799 fluorescence microscopy Methods 0.000 claims abstract description 17
- 229940024606 amino acid Drugs 0.000 claims abstract description 14
- 125000003277 amino group Chemical group 0.000 claims abstract description 10
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 claims abstract description 8
- 239000001257 hydrogen Substances 0.000 claims abstract description 8
- 229910052739 hydrogen Inorganic materials 0.000 claims abstract description 8
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 58
- 239000000243 solution Substances 0.000 claims description 41
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 claims description 33
- 238000006243 chemical reaction Methods 0.000 claims description 33
- 238000000502 dialysis Methods 0.000 claims description 33
- 239000000178 monomer Substances 0.000 claims description 29
- 229910052757 nitrogen Inorganic materials 0.000 claims description 29
- 239000008367 deionised water Substances 0.000 claims description 22
- 229910021641 deionized water Inorganic materials 0.000 claims description 22
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 15
- 239000007864 aqueous solution Substances 0.000 claims description 15
- 238000000034 method Methods 0.000 claims description 15
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 12
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 claims description 11
- 239000000203 mixture Substances 0.000 claims description 11
- YXMISKNUHHOXFT-UHFFFAOYSA-N (2,5-dioxopyrrolidin-1-yl) prop-2-enoate Chemical compound C=CC(=O)ON1C(=O)CCC1=O YXMISKNUHHOXFT-UHFFFAOYSA-N 0.000 claims description 10
- 239000003054 catalyst Substances 0.000 claims description 9
- 238000004090 dissolution Methods 0.000 claims description 9
- 238000004108 freeze drying Methods 0.000 claims description 9
- RDOXTESZEPMUJZ-UHFFFAOYSA-N anisole Chemical compound COC1=CC=CC=C1 RDOXTESZEPMUJZ-UHFFFAOYSA-N 0.000 claims description 8
- YNHIGQDRGKUECZ-UHFFFAOYSA-L bis(triphenylphosphine)palladium(ii) dichloride Chemical compound [Cl-].[Cl-].[Pd+2].C1=CC=CC=C1P(C=1C=CC=CC=1)C1=CC=CC=C1.C1=CC=CC=C1P(C=1C=CC=CC=1)C1=CC=CC=C1 YNHIGQDRGKUECZ-UHFFFAOYSA-L 0.000 claims description 8
- KZSNJWFQEVHDMF-BYPYZUCNSA-N L-valine Chemical compound CC(C)[C@H](N)C(O)=O KZSNJWFQEVHDMF-BYPYZUCNSA-N 0.000 claims description 7
- KZSNJWFQEVHDMF-UHFFFAOYSA-N Valine Natural products CC(C)C(N)C(O)=O KZSNJWFQEVHDMF-UHFFFAOYSA-N 0.000 claims description 7
- 239000004474 valine Substances 0.000 claims description 7
- ROHFNLRQFUQHCH-YFKPBYRVSA-N L-leucine Chemical compound CC(C)C[C@H](N)C(O)=O ROHFNLRQFUQHCH-YFKPBYRVSA-N 0.000 claims description 6
- ROHFNLRQFUQHCH-UHFFFAOYSA-N Leucine Natural products CC(C)CC(N)C(O)=O ROHFNLRQFUQHCH-UHFFFAOYSA-N 0.000 claims description 6
- KDLHZDBZIXYQEI-UHFFFAOYSA-N Palladium Chemical compound [Pd] KDLHZDBZIXYQEI-UHFFFAOYSA-N 0.000 claims description 6
- 238000001914 filtration Methods 0.000 claims description 6
- -1 succinimide ester Chemical class 0.000 claims description 6
- AGPKZVBTJJNPAG-WHFBIAKZSA-N L-isoleucine Chemical compound CC[C@H](C)[C@H](N)C(O)=O AGPKZVBTJJNPAG-WHFBIAKZSA-N 0.000 claims description 5
- 229960000310 isoleucine Drugs 0.000 claims description 5
- AGPKZVBTJJNPAG-UHFFFAOYSA-N isoleucine Natural products CCC(C)C(N)C(O)=O AGPKZVBTJJNPAG-UHFFFAOYSA-N 0.000 claims description 5
- 239000011259 mixed solution Substances 0.000 claims description 5
- 239000003960 organic solvent Substances 0.000 claims description 5
- UKODFQOELJFMII-UHFFFAOYSA-N pentamethyldiethylenetriamine Chemical compound CN(C)CCN(C)CCN(C)C UKODFQOELJFMII-UHFFFAOYSA-N 0.000 claims description 5
- UZKWTJUDCOPSNM-UHFFFAOYSA-N methoxybenzene Substances CCCCOC=C UZKWTJUDCOPSNM-UHFFFAOYSA-N 0.000 claims description 4
- 229910021589 Copper(I) bromide Inorganic materials 0.000 claims description 3
- 229910052763 palladium Inorganic materials 0.000 claims description 3
- 239000002904 solvent Substances 0.000 claims description 3
- 238000007112 amidation reaction Methods 0.000 claims description 2
- 238000010560 atom transfer radical polymerization reaction Methods 0.000 claims description 2
- 238000005859 coupling reaction Methods 0.000 claims description 2
- KZNICNPSHKQLFF-UHFFFAOYSA-N dihydromaleimide Natural products O=C1CCC(=O)N1 KZNICNPSHKQLFF-UHFFFAOYSA-N 0.000 claims description 2
- 238000001035 drying Methods 0.000 claims description 2
- 238000003756 stirring Methods 0.000 claims description 2
- 229960002317 succinimide Drugs 0.000 claims description 2
- 239000000463 material Substances 0.000 abstract description 10
- 230000003013 cytotoxicity Effects 0.000 abstract description 8
- 231100000135 cytotoxicity Toxicity 0.000 abstract description 8
- 150000005693 branched-chain amino acids Chemical class 0.000 abstract description 7
- 230000000694 effects Effects 0.000 abstract description 7
- 108090000623 proteins and genes Proteins 0.000 abstract description 6
- 102000004169 proteins and genes Human genes 0.000 abstract description 6
- 230000002159 abnormal effect Effects 0.000 abstract description 4
- 235000015097 nutrients Nutrition 0.000 abstract description 4
- 230000035755 proliferation Effects 0.000 abstract description 4
- 108010057904 polyisoleucine Proteins 0.000 abstract 1
- 108010033356 polyvaline Proteins 0.000 abstract 1
- 210000004027 cell Anatomy 0.000 description 14
- 238000010521 absorption reaction Methods 0.000 description 9
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 6
- 238000000862 absorption spectrum Methods 0.000 description 6
- 238000003384 imaging method Methods 0.000 description 6
- 201000011510 cancer Diseases 0.000 description 5
- 239000000975 dye Substances 0.000 description 5
- 206010006187 Breast cancer Diseases 0.000 description 4
- 208000026310 Breast neoplasm Diseases 0.000 description 4
- 238000002835 absorbance Methods 0.000 description 4
- 230000003833 cell viability Effects 0.000 description 4
- 230000005284 excitation Effects 0.000 description 4
- 239000002202 Polyethylene glycol Substances 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 238000003333 near-infrared imaging Methods 0.000 description 3
- VOFUROIFQGPCGE-UHFFFAOYSA-N nile red Chemical compound C1=CC=C2C3=NC4=CC=C(N(CC)CC)C=C4OC3=CC(=O)C2=C1 VOFUROIFQGPCGE-UHFFFAOYSA-N 0.000 description 3
- 239000002245 particle Substances 0.000 description 3
- 229920001223 polyethylene glycol Polymers 0.000 description 3
- UDGKZGLPXCRRAM-UHFFFAOYSA-N 1,2,5-thiadiazole Chemical compound C=1C=NSN=1 UDGKZGLPXCRRAM-UHFFFAOYSA-N 0.000 description 2
- XXSPGBOGLXKMDU-UHFFFAOYSA-M 2-bromo-2-methylpropanoate Chemical compound CC(C)(Br)C([O-])=O XXSPGBOGLXKMDU-UHFFFAOYSA-M 0.000 description 2
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 2
- 241000282414 Homo sapiens Species 0.000 description 2
- YTPLMLYBLZKORZ-UHFFFAOYSA-N Thiophene Chemical group C=1C=CSC=1 YTPLMLYBLZKORZ-UHFFFAOYSA-N 0.000 description 2
- 230000005540 biological transmission Effects 0.000 description 2
- 125000001246 bromo group Chemical group Br* 0.000 description 2
- 238000004113 cell culture Methods 0.000 description 2
- 238000002591 computed tomography Methods 0.000 description 2
- 238000010226 confocal imaging Methods 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 238000002296 dynamic light scattering Methods 0.000 description 2
- 238000000295 emission spectrum Methods 0.000 description 2
- 125000005843 halogen group Chemical group 0.000 description 2
- 239000003446 ligand Substances 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 238000000655 nuclear magnetic resonance spectrum Methods 0.000 description 2
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 description 2
- 229920001184 polypeptide Polymers 0.000 description 2
- 108090000765 processed proteins & peptides Proteins 0.000 description 2
- 102000004196 processed proteins & peptides Human genes 0.000 description 2
- 150000003384 small molecules Chemical class 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 230000002123 temporal effect Effects 0.000 description 2
- 210000001519 tissue Anatomy 0.000 description 2
- 238000005481 NMR spectroscopy Methods 0.000 description 1
- 108010087230 Sincalide Proteins 0.000 description 1
- 238000003917 TEM image Methods 0.000 description 1
- 230000002776 aggregation Effects 0.000 description 1
- 238000004220 aggregation Methods 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000010609 cell counting kit-8 assay Methods 0.000 description 1
- 210000003855 cell nucleus Anatomy 0.000 description 1
- 208000026106 cerebrovascular disease Diseases 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- NKNDPYCGAZPOFS-UHFFFAOYSA-M copper(i) bromide Chemical group Br[Cu] NKNDPYCGAZPOFS-UHFFFAOYSA-M 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 238000013399 early diagnosis Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 210000002919 epithelial cell Anatomy 0.000 description 1
- 239000007850 fluorescent dye Substances 0.000 description 1
- 208000019622 heart disease Diseases 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000011503 in vivo imaging Methods 0.000 description 1
- 238000011065 in-situ storage Methods 0.000 description 1
- 238000002595 magnetic resonance imaging Methods 0.000 description 1
- 239000005267 main chain polymer Substances 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 238000001000 micrograph Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 239000002105 nanoparticle Substances 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 239000012299 nitrogen atmosphere Substances 0.000 description 1
- 108091008104 nucleic acid aptamers Proteins 0.000 description 1
- 230000035515 penetration Effects 0.000 description 1
- 238000011056 performance test Methods 0.000 description 1
- 230000035699 permeability Effects 0.000 description 1
- 238000002600 positron emission tomography Methods 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- 238000004445 quantitative analysis Methods 0.000 description 1
- 238000006862 quantum yield reaction Methods 0.000 description 1
- 239000001044 red dye Substances 0.000 description 1
- 230000013878 renal filtration Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000007789 sealing 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
- 238000002603 single-photon emission computed tomography Methods 0.000 description 1
- 239000011949 solid catalyst Substances 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 125000003944 tolyl group Chemical group 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
- A61K49/001—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
- A61K49/0019—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08F—MACROMOLECULAR COMPOUNDS OBTAINED BY REACTIONS ONLY INVOLVING CARBON-TO-CARBON UNSATURATED BONDS
- C08F8/00—Chemical modification by after-treatment
- C08F8/30—Introducing nitrogen atoms or nitrogen-containing groups
- C08F8/32—Introducing nitrogen atoms or nitrogen-containing groups by reaction with amines
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Veterinary Medicine (AREA)
- Engineering & Computer Science (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Biomedical Technology (AREA)
- Epidemiology (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Medicinal Chemistry (AREA)
- Polymers & Plastics (AREA)
- Organic Chemistry (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Abstract
The invention discloses a tumor targeting water-soluble conjugated polymer contrast agent, a preparation method and application thereof, wherein the contrast agent has a chemical structure shown in the following formula III:wherein, -NH-R is an amino acid branched chain obtained by removing one hydrogen from an amino acid. The side chain of the tumor targeted water-soluble conjugated polymer contrast agent selects common branched chain amino acids in three proteins to provide nutrient substances required by abnormal proliferation of tumors, and is connected to the side end of a polymer through covalent bonds, and the polymer contains poly isoleucine, poly valine and poly brightlyThe amino acid not only can provide a large amount of amino groups, so that the near infrared two-region conjugated polymer has good water solubility, but also can realize tumor targeting, further reduce cytotoxicity, enhance the uptake of cells to materials, improve the enrichment rate of the materials, and further improve the effect of fluorescence imaging.
Description
Technical Field
The invention relates to the technical field of nano biomedical photosensitive diagnosis and treatment, in particular to a tumor targeting water-soluble conjugated polymer contrast agent, a preparation method and application thereof.
Background
Malignant tumor is also called cancer, is one of the greatest threats facing human beings at present, and is the first three diseases which lead to death of human beings due to diseases together with heart diseases and cerebrovascular diseases. Malignant tumors have high mortality due to the characteristics of various types, high complexity, hidden early symptoms and the like. Thus, early diagnosis of cancer has become a popular research direction.
In recent years, various imaging techniques, including Computed Tomography (CT), magnetic Resonance Imaging (MRI), and Positron Emission Tomography (PET) and Single Photon Emission Computed Tomography (SPECT), have been used for in vivo imaging. However, simultaneous in-situ, real-time imaging of multiple events in a living body is difficult to achieve due to insufficient spatial and temporal resolution and safety concerns. Fluorescence imaging of the second near infrared window (NIR-II, 1000-1700 nm) is an emerging technology with deeper tissue penetration, higher spatial resolution and higher temporal resolution than traditional fluorescence imaging (400-900 nm) because it reduces photon absorption and scattering and ignores tissue autofluorescence.
Traditional targeting strategies include passive targeting, which promotes selective aggregation in tumors by modulating nanoparticle size and surface chemistry, enhancing Permeability and Retention (EPR) effects, and active targeting, which has the disadvantages of limited enrichment and high particle size requirements. Active targeting is through binding of high affinity ligands including polypeptides, proteins, nucleic acid aptamers, etc. to specific surface molecules that are predominantly expressed by cancer cells or tumor epithelial cells, with the disadvantage of relatively high costs for polypeptide proteins, etc. The improvement of enrichment efficiency is critical to improve contrast agent specific targeting. Traditional contrast agents are prepared by coating contrast agents in polyethylene glycol (PEG) to enter cells, targeting tumors by EPR effect, and the polyethylene glycol with neutral water-soluble groups has no specific recognition capability with other molecules or structures, so the enrichment degree is limited. And so far only a few organic molecules, which are highly hydrophobic, water-insoluble dyes, have fluorescent emission in the NIR-II region, which must be encapsulated in a polymer matrix for bioimaging, increasing the particle size beyond the renal filtration threshold (ca.40 kd). Recently, small molecule CH1055 (8.9 kDa) NIR-II organic dyes emit at about 1055nm under 808nm excitation, with high water solubility. Although this small molecule dye has shown promise in the preclinical, the study of NIR-II organic dyes with simple preparation process, high quantum yield, and high resolution imaging quality is still in the beginning.
Disclosure of Invention
The invention aims to solve the technical problem of providing a tumor targeting water-soluble conjugated polymer contrast agent, a preparation method and application thereof aiming at the defects in the prior art.
In order to solve the technical problems, the invention adopts the following technical scheme: in a first aspect of the present invention, there is provided a tumor-targeted water-soluble conjugated polymer contrast agent having a chemical structure represented by the following formula III:
wherein, -NH-R is an amino acid side chain obtained by removing one hydrogen from an amino group in an amino acid, x=y=4-8, and m=30-50.
The targeted water-soluble conjugated polymer contrast agent has an amphiphilic molecular structure and comprises a polymer main chain and a side chain, wherein the polymer main chain has a donor and acceptor structure, and can prolong the length of a conjugated bond in the conjugated skeleton, so that the absorption is red-shifted, the fluorescence intensity of the polymer is improved, and the living body contrast resolution and contrast are improved; the side chains select common branched chain amino acids in three proteins to provide nutrients required by abnormal proliferation of tumors, and the branched chain amino acids are connected to the side ends of the polymers through covalent bonds, so that a large number of amino groups can be provided, the near infrared two-region conjugated polymer has good water solubility, more importantly, tumor targeting can be realized, cytotoxicity can be further reduced, the uptake degree of cells to the materials can be enhanced, the enrichment rate of the materials can be improved, and the fluorescence imaging effect can be further improved. The polymer has the characteristics, so that the polymer has the advantages of improving focus targeting enrichment, signal-to-noise ratio, imaging definition and the like when being used for fluorescence imaging.
The end sealing group of the structure shown in the formula I and the thiophene ring isThe end-capping group attached to the benzene ring is a halo group, such as bromo.
In a second aspect of the present invention, there is provided a method for preparing a tumor-targeted water-soluble conjugated polymer contrast agent as described above, comprising the steps of:
s1, preparing a polymer 1 with a structure shown in the following formula I:
wherein x=y=4 to 8;
s2, carrying out atom transfer radical polymerization reaction on the polymer 1 and acrylic acid-N-succinimidyl ester to prepare a polymer 2 with a structure shown in the following formula II:
wherein m=30 to 50;
s3, removing succinimide ester from the polymer 2, and carrying out amidation reaction with amino groups of amino acids to prepare the polymer 3, namely the tumor targeting water-soluble conjugated polymer contrast agent, wherein the structural formula of the polymer is shown in a formula III.
Wherein the amino acid is any one of isoleucine, valine and leucine.
The targeted water-soluble conjugated polymer contrast agent has an amphiphilic molecular structure and comprises a polymer main chain and a side chain, wherein the polymer main chain has a donor and acceptor structure, and can prolong the length of a conjugated bond in the conjugated skeleton, so that the absorption is red-shifted, the fluorescence intensity of the polymer is improved, and the living body contrast resolution and contrast are improved; the side chains select common branched chain amino acids in three proteins to provide nutrients required by abnormal proliferation of tumors, and the branched chain amino acids are connected to the side ends of the polymers through covalent bonds, so that a large number of amino groups can be provided, the near infrared two-region conjugated polymer has good water solubility, more importantly, tumor targeting can be realized, cytotoxicity can be further reduced, the uptake degree of cells to the materials can be enhanced, the enrichment rate of the materials can be improved, and the fluorescence imaging effect can be further improved. The polymer has the characteristics, so that the polymer has the advantages of improving focus targeting enrichment, signal-to-noise ratio, imaging definition and the like when being used for fluorescence imaging.
For example, in the structure shown in formula II, the end capping group attached to the thiophene ring isThe end-capping group attached to the benzene ring is a halo group, such as bromo.
Preferably, in the step S1, the polymer 1 is prepared by carrying out a steller coupling reaction on a monomer 1 having a structure shown in formula iv, a monomer 2 having a structure shown in formula V, and a monomer 3 having a structure shown in formula vi, and the synthetic route is as follows:
specifically, monomer 1 is: 4, 7-dibromobenzo [1,2-c:4,5-c' ] bis ([ 1,2,5] thiadiazole, structural formula IV;
monomer 2 is: 2, 6-bis (trimethyltin) -4, 8-bis (5- (2-ethylhexyl) thiophen-2-yl) -benzodithiophene having formula V;
monomer 3 is: (2, 7-dibromo-9H-fluorene-9, 9-diyl) bis (propane-3, 1-diyl) bis (2-bromo-2-methylpropionate) having the structural formula VI.
Preferably, the step S1 specifically includes: and under the dark environment, adding the monomer 1, the monomer 2 and the monomer 3 into a solvent to be completely dissolved, then introducing nitrogen into the obtained solution to bubble for more than 20 minutes, adding a palladium catalyst into the solution, reacting for 2-4 hours at the temperature of 80-90 ℃ under the protection of nitrogen, filtering the product after the reaction is finished, and then settling in methanol to obtain the polymer 1.
Preferably, in the step S1, monomer 1: monomer 2: the molar ratio of the monomer 3 to the feed is 1:2:1.
preferably, the step S1 specifically includes: under a dark environment, 0.2mM of monomer 1, 0.2mM of monomer 2 and 0.1mM of monomer 3 are added into 1mL of toluene to be completely dissolved, then nitrogen is introduced into the obtained solution to be bubbled for more than 20 minutes, 50mg of bis (triphenylphosphine) palladium (II) dichloride is added into the solution, the reaction is carried out for 3 hours under the protection of nitrogen at 80 ℃, and after the reaction is finished, the product is filtered and then repeatedly settled in methanol, so that the polymer 1 is obtained. Among them, the palladium catalyst is preferably bis (triphenylphosphine) palladium (II) dichloride, and the solvent is preferably toluene.
Preferably, the step S2 specifically includes:
adding a polymer 1 and acrylic acid-N-succinimidyl ester into an organic solvent under the condition of avoiding light, then adding a catalyst, adding pentamethyldiethylenetriamine under the protection of nitrogen, reacting the obtained mixed solution for 9-12 hours at 80-100 ℃, filtering a product after the reaction is finished, and then repeatedly settling in diethyl ether to obtain a polymer 2; the synthetic route is as follows:
preferably, the molar ratio of polymer 1 to N-succinimidyl acrylate is between 1:50 and 200, for example 1:100.
Preferably, the catalyst is cuprous bromide and the organic solvent is anisole.
Preferably, the step S2 specifically includes:
100mg of polymer 1, 887mg of acrylic acid-N-succinimidyl ester are added into 1mL of anisole under the dark condition, then a catalyst CuBr is added, 50 mu L of pentamethyldiethylenetriamine is added under the protection of nitrogen, the obtained mixed solution is reacted for 12 hours at 90 ℃, and after the reaction is finished, the product is filtered, and then the mixture is repeatedly settled in diethyl ether, so that polymer 2 is obtained.
Preferably, the step S3 specifically includes:
under the condition of avoiding light, adding polymer 2 and amino acid into DMF for dissolution, adding triethylamine under the protection of nitrogen, and stirring and reacting for 24-60h at 35-45 ℃; and after the reaction is finished, adding the solution in the product system into deionized water, placing the obtained mixed aqueous solution into a dialysis bag, dialyzing with deionized water, and drying the solution in the dialysis bag after the dialysis is finished to obtain the polymer 3.
Preferably, the step S3 specifically includes:
300mg of polymer 2 and 1.75mmol of isoleucine are dissolved in 5mL of DMF under the protection of nitrogen, 1.75mmol of triethylamine is added under the protection of nitrogen, and the mixture is stirred at 40 ℃ for reaction for 48 hours; adding the solution in the product system into 20mL of deionized water after the reaction is finished, placing the obtained mixed aqueous solution into a dialysis bag with 3500 molecular weight cut-off, dialyzing with deionized water for 48h, changing the dialyzate for 1 time every 6h, and freeze-drying the solution in the dialysis bag after the dialysis is finished to obtain a polymer 3 with a chemical structure shown in a formula III-1; the synthetic route is as follows:
preferably, the step S3 specifically includes:
under the protection of light, 300mg of polymer 2 and 1.75mmol of valine are added into 5mL of DMF for dissolution, 1.75mmol of triethylamine is added under the protection of nitrogen, and the mixture is stirred at 40 ℃ for reaction for 48h; adding the solution in the product system into 20mL of deionized water after the reaction is finished, placing the obtained mixed aqueous solution into a dialysis bag with 3500 molecular weight cut-off, dialyzing with deionized water for 48h, changing the dialyzate for 1 time every 6h, and freeze-drying the solution in the dialysis bag after the dialysis is finished to obtain a polymer 3 with a chemical structure shown in a formula III-2; the synthetic route is as follows:
preferably, the step S3 specifically includes:
300mg of polymer 2 and 1.75mmol of leucine are added into 5mL of DMF for dissolution under the protection of nitrogen, 1.75mmol of triethylamine is added under the protection of nitrogen, and the mixture is stirred at 40 ℃ for reaction for 48h; adding the solution in the product system into 20mL of deionized water after the reaction is finished, placing the obtained mixed aqueous solution into a dialysis bag with 3500 molecular weight cut-off, dialyzing with deionized water for 48h, changing the dialyzate for 1 time every 6h, and freeze-drying the solution in the dialysis bag after the dialysis is finished to obtain a polymer 3 with a chemical structure shown in a formula III-3; the synthetic route is as follows:
in a third aspect of the invention, the application of the tumor-targeted water-soluble conjugated polymer contrast agent in near infrared second window fluorescence imaging is provided.
The beneficial effects of the invention are as follows:
the tumor targeted water-soluble conjugated polymer contrast agent provided by the invention has an amphiphilic molecular structure and contains a polymer main chain and a side chain, wherein the polymer main chain has a donor and acceptor structure, so that the conjugated bond length in the conjugated skeleton can be prolonged, the absorption is red-shifted, the fluorescence intensity of the polymer is improved, and the living body contrast resolution and contrast are improved; the side chains select common branched chain amino acids in three proteins to provide nutrients required by abnormal proliferation of tumors, and the branched chain amino acids are connected to the side ends of the polymers through covalent bonds, so that a large number of amino groups can be provided, the near infrared two-region conjugated polymer has good water solubility, more importantly, tumor targeting can be realized, cytotoxicity can be further reduced, the uptake degree of cells to the materials can be enhanced, the enrichment rate of the materials can be improved, and the fluorescence imaging effect can be further improved. The polymer has the advantages of improving focus targeting enrichment, signal to noise ratio, imaging definition and the like when being used for fluorescence imaging, so that the polymer can be well applied to near infrared second window fluorescence imaging.
Drawings
FIG. 1 is an absorption spectrum of a polymer 1 prepared in example 1;
FIG. 2 is an absorption spectrum of the polymer 2 prepared in example 1;
FIG. 3 is a graph showing the hydrogen nuclear magnetic resonance spectra of the polymers 3-1, 3-2 and 3-3 obtained in examples 2 to 4;
FIG. 4 is a transmission electron micrograph of the polymers 3-1, 3-2 and 3-3 prepared in examples 2 to 4;
FIG. 5 is a schematic diagram showing the nano-diameter measured by dynamic light scattering of the polymers 3-1, 3-2, and 3-3 prepared in examples 2 to 4;
FIG. 6 is a graph showing absorption and emission spectra of the polymers 3-1, 3-2 and 3-3 (1 mg/mL) obtained in examples 2 to 4;
FIG. 7 is a view showing the cell confocal imaging of the polymers 3-1, 3-2 and 3-3 produced in examples 2 to 4;
FIG. 8 is a graph showing quantitative analysis of fluorescence intensity of cell confocal imaging patterns of polymers 3-1, 3-2 and 3-3 obtained in examples 2 to 4;
FIG. 9 is a second window near infrared and fluorescence imaging of the targeted aqueous conjugated polymer contrast agent solution prepared in example 2;
FIG. 10 is a second window near infrared and fluorescence imaging of the targeted aqueous solution of the conjugated polymer contrast agent prepared in example 3;
FIG. 11 is a second window near infrared and fluorescence imaging of the targeted aqueous solution of the conjugated polymer contrast agent prepared in example 4;
FIG. 12 is a graph showing the cell viability of the targeted water-soluble conjugated polymer prepared in example 2 with respect to cytotoxicity of mouse breast cancer cells;
FIG. 13 is a graph showing the cell viability of the targeted water-soluble conjugated polymer prepared in example 3 against cytotoxicity of mouse breast cancer cells;
FIG. 14 is a graph showing the cell viability of the targeted water-soluble conjugated polymer prepared in example 4 against cytotoxicity of mouse breast cancer cells.
Detailed Description
The present invention is described in further detail below with reference to examples to enable those skilled in the art to practice the same by referring to the description.
It will be understood that terms, such as "having," "including," and "comprising," as used herein, do not preclude the presence or addition of one or more other elements or groups thereof.
The test methods used in the following examples are conventional methods unless otherwise specified. The material reagents and the like used in the following examples are commercially available unless otherwise specified. The following examples were conducted under conventional conditions or conditions recommended by the manufacturer, without specifying the specific conditions. The reagents or apparatus used were conventional products commercially available without the manufacturer's attention.
Example 1 preparation of Polymer 1, polymer 2
1. The chemical structure of polymer 1 is as follows:
wherein x=y=4 to 8;
the preparation method comprises the following steps:
in a dark environment, 4, 7-dibromobenzo [1,2-c:4,5-c' ] bis ([ 1,2,5] thiadiazole (35.2 mg,0.2 mM), donor molecule (monomer 2) 2, 6-bis (trimethyltin) -4, 8-bis (5- (2-ethylhexyl) thiophen-2-yl) -benzodithiophene (180.9 mg,0.2 mM), donor molecule (monomer 3) (2, 7-dibromo-9H-fluorene-9, 9-diyl) bis (propane-3, 1-diyl) bis (2-bromo-2-methylpropionate) (73.8 mg,0.1 mM) were added to the reaction tube, after dissolving the above solid with toluene (1 mL), nitrogen was bubbled for 20 minutes or more, bis (triphenylphosphine) palladium (II) dichloride (50 mg) was rapidly added to the flask, and reacted under nitrogen protection at 80℃for 3 hours, and the crude polymer was filtered with a filter membrane and the brown polymer was removed by repeating the filtration to obtain a flask (100 mg).
2. The chemical structure of polymer 2 is as follows:
wherein m=30 to 50;
the preparation method comprises the following steps:
polymer 1 (100 mg), acrylic acid-N-succinimidyl ester (887 mg,5.24 mM) was added to a reaction tube under a dark condition, the above solid was dissolved with anisole (1 mL), cuBr was added as a catalyst, 50. Mu.L of pentamethyldiethylenetriamine was added as a ligand under a nitrogen atmosphere, the resulting mixed solution was reacted at 90℃for 12 hours, after the completion of the reaction, the solid catalyst was removed by filtration with an organic filter membrane, and then repeatedly settled in diethyl ether to obtain a reddish-brown acrylic acid-N-succinimidyl ester side chain-substituted conjugated main chain polymer, polymer 2 (900 mg).
Polymer 1 and Polymer 2 in examples 2 to 4 below were prepared by this example.
Example 2
A tumor targeting water-soluble conjugated polymer contrast agent has a chemical structure shown in the following formula III-1:
wherein x=y=4 to 8, and m=30 to 50.
wherein-NH-R is an amino acid branched chain obtained by removing one hydrogen from an amino group in isoleucine.
The preparation method comprises the following steps: under the condition of avoiding light, 300mg of polymer 2 and 1.75mmol of valine are added into 5mL of DMF for dissolution, 1.75mmol of triethylamine is added under the protection of nitrogen, and the mixture is stirred at 40 ℃ for reaction for 48 hours; adding the solution in the product system into 20mL of deionized water after the reaction is finished, placing the obtained mixed aqueous solution into a dialysis bag with 3500 molecular weight cut-off, dialyzing with deionized water for 48h, changing the dialyzate for 1 time every 6h, and freeze-drying the solution in the dialysis bag after the dialysis is finished to obtain a polymer with a chemical structure shown as a formula III-2, and marking the polymer as a polymer 3-1; the synthetic route is as follows:
example 3
A tumor targeting water-soluble conjugated polymer contrast agent has a chemical structure shown in the following formula III-2:
wherein x=y=4 to 8, and m=30 to 50.
wherein-NH-R is an amino acid branched chain obtained by removing one hydrogen from an amino group in valine.
The preparation method comprises the following steps:
under the condition of avoiding light, 300mg of polymer 2 and 1.75mmol of valine are added into 5mL of DMF for dissolution, 1.75mmol of triethylamine is added under the protection of nitrogen, and the mixture is stirred at 40 ℃ for reaction for 48 hours; adding the solution in the product system into 20mL of deionized water after the reaction is finished, placing the obtained mixed aqueous solution into a dialysis bag with 3500 molecular weight cut-off, dialyzing with deionized water for 48h, changing the dialyzate for 1 time every 6h, and freeze-drying the solution in the dialysis bag after the dialysis is finished to obtain a polymer with a chemical structure shown as a formula III-2, and marking the polymer as a polymer 3-2; the synthetic route is as follows:
example 4
A tumor targeting water-soluble conjugated polymer contrast agent has a chemical structure shown in the following formula III-3:
wherein x=y=4 to 8, and m=30 to 50.
wherein-NH-R is an amino acid branched chain obtained by removing one hydrogen from an amino group in leucine.
The preparation method comprises the following steps:
under the protection of light, 300mg of polymer 2 and 1.75mmol of leucine are added into 5mL of DMF for dissolution, 1.75mmol of triethylamine is added under the protection of nitrogen, and the mixture is stirred at 40 ℃ for reaction for 48 hours; adding the solution in the product system into 20mL of deionized water after the reaction is finished, placing the obtained mixed aqueous solution into a dialysis bag with 3500 molecular weight cut-off, dialyzing with deionized water for 48h, changing the dialyzate for 1 time every 6h, and freeze-drying the solution in the dialysis bag after the dialysis is finished to obtain a polymer with a chemical structure shown as a formula III-3, and marking the polymer as a polymer 3-3; the synthetic route is as follows:
the following performance tests and characterization were performed on the products prepared in examples 1-4:
1. the organic solution of the polymer 1 prepared in example 1 was prepared using methylene chloride as an organic solvent, and the absorption spectrum was tested, and as shown in fig. 1, it was seen that the absorption peak was located in the 734 nm band.
2. The organic solution of the polymer 2 prepared in example 1 was prepared using methylene chloride as an organic solvent, and the absorption spectrum was tested, and as shown in FIG. 2, it was seen that the absorption peak was located in the 727 nm band.
3. The tumor-targeted water-soluble conjugated polymer contrast agents (sequentially denoted as polymer 3-1, polymer 3-2 and polymer 3-3) prepared in examples 2 to 4 were subjected to nuclear magnetic resonance spectrum detection to obtain nuclear magnetic resonance hydrogen spectrograms of the targeted water-soluble conjugated polymer contrast agents, and characteristic proton signals of amino acid side chains can be seen as shown in fig. 3.
4. The tumor-targeted water-soluble conjugated polymer contrast agents (sequentially designated as Polymer 3-1, polymer 3-2, polymer 3-3) prepared in examples 2 to 4 were prepared into aqueous polymer solutions having a concentration of 1mg/mL, respectively, and then the following tests were performed:
i) The aqueous solutions were analyzed by a transmission electron microscope, and the obtained transmission electron microscope image was shown in FIG. 4. As can be seen from FIG. 4, the particle sizes of the polymer 3-1, the polymer 3-2 and the polymer 3-3 were all about 100 nm.
ii) based on the formulated aqueous polymer solution, measured according to dynamic light scattering: the average hydrodynamic diameter of polymer 3-1 in water was about 102 nm, the average hydrodynamic diameter of polymer 3-2 in water was about 90 nm, and the average hydrodynamic diameter of polymer 3-3 in water was about 90 nm, and the resulting hydrodynamic volume diagrams of the molecules are shown in FIG. 5.
iii) Based on the absorption and emission spectra of the polymers 3-1, 3-2 and 3-3, respectively, based on the aqueous polymer solution, it can be seen that the results are shown in FIG. 6: the absorption peak of the polymer 3-1 is positioned in 970 nanometer wave band, and the emission peak is positioned in 1047 nanometer wave band under 808 nanometer laser excitation; the absorption peak of the polymer 3-2 is positioned in a 958 nanometer wave band, and the emission peak of the polymer is positioned in a 1048 nanometer wave band under 808 nanometer laser excitation; the absorption peak of the polymer 3-3 is located in the 926 nanometer wave band, and the emission peak of the polymer under 808 nanometer laser excitation is located in the 1069 nanometer wave band.
5. Polymer 3-1, polymer 3-2 and Polymer 3-3 prepared in examples 2 to 4 were prepared as aqueous solutions of polymers having concentration gradients of 2mg/mL, 1mg/mL, 0.5mg/mL and 0.25mg/mL, respectively, and excited with 1064 nm laser light, and the fluorescence imaging images of the solutions were taken in a second near infrared window fluorescence imager, and the results are shown in FIGS. 9 to 11, which illustrate that Polymer 3-1, polymer 3-2 and Polymer 3-3 all have near infrared two-region fluorescence imaging properties.
6. 10mg of each of the polymer 3-1, the polymer 3-2 and the polymer 3-3 prepared in examples 2 to 4 was dissolved in a small amount of dimethyl sulfoxide solution, and the nile red dye was coated by a nano precipitation method, and the absorbance of nile red at 552nm was detected to be the same absorbance, and the fluorescence effect of the dye was observed by a confocal microscope. Fluorescence intensity was calculated and the uptake of different materials by the cells was compared. The confocal microscope shooting result is shown in fig. 7, wherein a cell nucleus fluorescent dye fluorescent image, a nile red fluorescent image and a superposed fluorescent image are sequentially shown from top to bottom; the polymer 3-1, the polymer 3-2 and the polymer 3-3 are arranged from left to right. It can be seen that: the fluorescence intensity of the polymer 3-1, the polymer 3-2 and the polymer 3-3 is high, which indicates that the targeted water-soluble conjugated polymer contrast agent prepared by the invention has high focus enrichment degree and good cell targeting. Quantitative average fluorescence intensity analysis is shown in fig. 8, and it can be seen that: the cell uptake rate of Polymer 3-3 was maximal.
7. Preparing a series of cell culture solutions with a certain concentration gradient from the polymers 3-1, 3-2 and 3-3 prepared in the examples 2-4 respectively, wherein the highest concentration of the polymers in the cell culture solution is 2mg/mL; the three culture solutions were used to culture with mouse breast cancer cells for 4 hours, CCK-8 reagent was added, absorbance at 450 nm was measured, and the obtained absorbance was converted into cell viability, as shown in FIGS. 12 to 14, and it was seen that: the cytotoxicity of the polymer 3-1, the polymer 3-2 and the polymer 3-3 is very low, which proves that the targeted water-soluble conjugated polymer contrast agent prepared by the invention has good biocompatibility.
Although embodiments of the present invention have been disclosed above, it is not limited to the use of the description and embodiments, it is well suited to various fields of use for the invention, and further modifications may be readily apparent to those skilled in the art, and accordingly, the invention is not limited to the particular details without departing from the general concepts defined in the claims and the equivalents thereof.
Claims (14)
1. The tumor targeting water-soluble conjugated polymer contrast agent is characterized by having a chemical structure shown in the following formula III:
wherein, -NH-R is an amino acid side chain obtained by removing one hydrogen from an amino acid, x=y=4-8, and m=30-50.
2. A method for preparing a tumor-targeted water-soluble conjugated polymer contrast agent according to claim 1, comprising the following steps:
s1, preparing a polymer 1 with a structure shown in the following formula I:
wherein x=y=4 to 8;
s2, carrying out atom transfer radical polymerization reaction on the polymer 1 and acrylic acid-N-succinimidyl ester to prepare a polymer 2 with a structure shown in the following formula II:
wherein m=30 to 50;
s3, removing succinimide ester from the polymer 2, and carrying out amidation reaction with amino groups of amino acids to prepare the polymer 3, namely the tumor targeting water-soluble conjugated polymer contrast agent, wherein the structural formula of the polymer is shown in a formula III.
Wherein the amino acid is any one of isoleucine, valine and leucine.
3. The method for preparing the tumor-targeted water-soluble conjugated polymer contrast agent according to claim 2, wherein in the step S1, the polymer 1 is prepared by carrying out a steller coupling reaction on a monomer 1 having a structure shown in formula iv, a monomer 2 having a structure shown in formula V and a monomer 3 having a structure shown in formula vi, and the synthetic route is as follows:
4. the method for preparing a tumor-targeted water-soluble conjugated polymer contrast agent according to claim 3, wherein the step S1 specifically comprises: and under the dark environment, adding the monomer 1, the monomer 2 and the monomer 3 into a solvent to be completely dissolved, then introducing nitrogen into the obtained solution to bubble for more than 20 minutes, adding a palladium catalyst into the solution, reacting for 2-4 hours at the temperature of 80-90 ℃ under the protection of nitrogen, filtering the product after the reaction is finished, and then settling in methanol to obtain the polymer 1.
5. The method for preparing a tumor-targeted water-soluble conjugated polymer contrast agent according to claim 4, wherein in the step S1, monomer 1: monomer 2: the molar ratio of the monomer 3 to the feed is 1:2:1.
6. the method for preparing a tumor-targeted water-soluble conjugated polymer contrast agent according to claim 5, wherein the step S1 specifically comprises: under a dark environment, 0.2mM of monomer 1, 0.2mM of monomer 2 and 0.1mM of monomer 3 are added into 1mL of toluene to be completely dissolved, then nitrogen is introduced into the obtained solution to be bubbled for more than 20 minutes, 50mg of bis (triphenylphosphine) palladium (II) dichloride is added into the solution, the reaction is carried out for 3 hours under the protection of nitrogen at 80 ℃, and after the reaction is finished, the product is filtered and then repeatedly settled in methanol, so that the polymer 1 is obtained.
7. The method for preparing a tumor-targeted water-soluble conjugated polymer contrast agent according to claim 3, wherein the step S2 specifically comprises:
adding a polymer 1 and acrylic acid-N-succinimidyl ester into an organic solvent under the condition of avoiding light, then adding a catalyst, adding pentamethyldiethylenetriamine under the protection of nitrogen, reacting the obtained mixed solution for 9-12 hours at 80-100 ℃, filtering a product after the reaction is finished, and then repeatedly settling in diethyl ether to obtain a polymer 2; the synthetic route is as follows:
8. the method for preparing a tumor-targeted water-soluble conjugated polymer contrast agent according to claim 7, wherein the molar ratio of the polymer 1 to the acrylic acid-N-succinimidyl ester is 1:50-200.
9. The method for preparing a tumor-targeted water-soluble conjugated polymer contrast agent according to claim 8, wherein the step S2 specifically comprises:
100mg of polymer 1, 887mg of acrylic acid-N-succinimidyl ester are added into 1mL of anisole under the dark condition, then a catalyst CuBr is added, 50 mu L of pentamethyldiethylenetriamine is added under the protection of nitrogen, the obtained mixed solution is reacted for 12 hours at 90 ℃, and after the reaction is finished, the product is filtered, and then the mixture is repeatedly settled in diethyl ether, so that polymer 2 is obtained.
10. The method for preparing a tumor-targeted water-soluble conjugated polymer contrast agent according to claim 2, wherein the step S3 specifically comprises:
under the condition of avoiding light, adding polymer 2 and amino acid into DMF for dissolution, adding triethylamine under the protection of nitrogen, and stirring and reacting for 24-60h at 35-45 ℃; and after the reaction is finished, adding the solution in the product system into deionized water, placing the obtained mixed aqueous solution into a dialysis bag, dialyzing with deionized water, and drying the solution in the dialysis bag after the dialysis is finished to obtain the polymer 3.
11. The method for preparing a tumor-targeted water-soluble conjugated polymer contrast agent according to claim 10, wherein the step S3 specifically comprises:
300mg of polymer 2 and 1.75mmol of isoleucine are dissolved in 5mL of DMF under the protection of nitrogen, 1.75mmol of triethylamine is added under the protection of nitrogen, and the mixture is stirred at 40 ℃ for reaction for 48 hours; adding the solution in the product system into 20mL of deionized water after the reaction is finished, placing the obtained mixed aqueous solution into a dialysis bag with 3500 molecular weight cut-off, dialyzing with deionized water for 48h, changing the dialyzate for 1 time every 6h, and freeze-drying the solution in the dialysis bag after the dialysis is finished to obtain a polymer 3 with a chemical structure shown in a formula III-1; the synthetic route is as follows:
12. the method for preparing a tumor-targeted water-soluble conjugated polymer contrast agent according to claim 10, wherein the step S3 specifically comprises:
under the protection of light, 300mg of polymer 2 and 1.75mmol of valine are added into 5mL of DMF for dissolution, 1.75mmol of triethylamine is added under the protection of nitrogen, and the mixture is stirred at 40 ℃ for reaction for 48h; adding the solution in the product system into 20mL of deionized water after the reaction is finished, placing the obtained mixed aqueous solution into a dialysis bag with 3500 molecular weight cut-off, dialyzing with deionized water for 48h, changing the dialyzate for 1 time every 6h, and freeze-drying the solution in the dialysis bag after the dialysis is finished to obtain a polymer 3 with a chemical structure shown in a formula III-2; the synthetic route is as follows:
13. the method for preparing a tumor-targeted water-soluble conjugated polymer contrast agent according to claim 10, wherein the step S3 specifically comprises:
300mg of polymer 2 and 1.75mmol of leucine are added into 5mL of DMF for dissolution under the protection of nitrogen, 1.75mmol of triethylamine is added under the protection of nitrogen, and the mixture is stirred at 40 ℃ for reaction for 48h; adding the solution in the product system into 20mL of deionized water after the reaction is finished, placing the obtained mixed aqueous solution into a dialysis bag with 3500 molecular weight cut-off, dialyzing with deionized water for 48h, changing the dialyzate for 1 time every 6h, and freeze-drying the solution in the dialysis bag after the dialysis is finished to obtain a polymer 3 with a chemical structure shown in a formula III-3; the synthetic route is as follows:
14. use of the tumor-targeted water-soluble conjugated polymer contrast agent according to claim 1 or the tumor-targeted water-soluble conjugated polymer contrast agent prepared by the method according to any one of claims 2 to 13 in near infrared second window fluorescence imaging.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310734976.0A CN116903771A (en) | 2023-06-20 | 2023-06-20 | Tumor targeting water-soluble conjugated polymer contrast agent, preparation method and application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310734976.0A CN116903771A (en) | 2023-06-20 | 2023-06-20 | Tumor targeting water-soluble conjugated polymer contrast agent, preparation method and application |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN116903771A true CN116903771A (en) | 2023-10-20 |
Family
ID=88361878
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202310734976.0A Pending CN116903771A (en) | 2023-06-20 | 2023-06-20 | Tumor targeting water-soluble conjugated polymer contrast agent, preparation method and application |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN116903771A (en) |
-
2023
- 2023-06-20 CN CN202310734976.0A patent/CN116903771A/en active Pending
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Chen et al. | Fabrication of fluorescent nanoparticles based on AIE luminogens (AIE dots) and their applications in bioimaging | |
| Chen et al. | Design and development of fluorescent nanostructures for bioimaging | |
| Zhang et al. | Aggregation induced emission-based fluorescent nanoparticles: fabrication methodologies and biomedical applications | |
| Li et al. | Polymer encapsulated conjugated polymer nanoparticles for fluorescence bioimaging | |
| Yu et al. | Near-infrared fluorescence imaging using organic dye nanoparticles | |
| CN101695476B (en) | Method for preparing medical nanoparticles | |
| CN110128665B (en) | Amphiphilic block polymer near-infrared fluorescent probe based on azo reductase response and application | |
| CN110960697B (en) | Preparation method of zwitterion-modified dendrimer-coated copper sulfide nanoparticle/pDNA (deoxyribonucleic acid) compound | |
| CN114057773A (en) | Near-infrared two-region aggregation-induced emission molecule and application thereof | |
| CN109529059A (en) | A kind of fluorescence-magnetic resonance bimodal quantum dot and its methods for making and using same | |
| CN113502159B (en) | Preparation method of pH activated near-infrared I region fluorescence-emitting carbon quantum dot, product and application thereof | |
| Zhang et al. | Organic-to-aqueous phase transfer of Zn–Cu–In–Se/ZnS quantum dots with multifunctional multidentate polymer ligands for biomedical optical imaging | |
| An et al. | A reticuloendothelial system-stealthy dye–albumin nanocomplex as a highly biocompatible and highly luminescent nanoprobe for targeted in vivo tumor imaging | |
| Guo et al. | Rapid synthesis of amphiphilic europium complexes via ultrasonic treatment-assisted crosslinking reaction | |
| CN113736456A (en) | Tumor targeting nanoprobe based on folic acid coupled carbon quantum dots and preparation method thereof | |
| US20110318275A1 (en) | Multifunctional contrast agent using biocompatible polymer and preparation method | |
| CN116903771A (en) | Tumor targeting water-soluble conjugated polymer contrast agent, preparation method and application | |
| CN113912762B (en) | Water-soluble near-infrared two-region macromolecular fluorescent probe and preparation method and application thereof | |
| Cheng et al. | A perylenediimide-based nanocarrier monitors curcumin release with an “off–on” fluorescence switch | |
| CN111693495A (en) | Near-infrared double-channel fluorescence living body microscopic imaging method | |
| CN116769174A (en) | Water-soluble conjugated polymer contrast agent, preparation method and application thereof | |
| CN118063708A (en) | Saccharide modified water-soluble conjugated polymer contrast agent, preparation method and application thereof | |
| CN111892645B (en) | Organic coordination compound, preparation method and application thereof, and probe | |
| CN111687428B (en) | Amphiphilic polymer mediated gold nanoparticle controllable assembly and preparation and application thereof | |
| CN113786492A (en) | Polymer carrier for photodynamic therapy and preparation method and application thereof |
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 |