CN105859807A - Beta-1,4-galactosyltransferase I substrate fluorescent molecular probe, and intermediate and preparation method thereof - Google Patents
Beta-1,4-galactosyltransferase I substrate fluorescent molecular probe, and intermediate and preparation method thereof Download PDFInfo
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- CN105859807A CN105859807A CN201610225439.3A CN201610225439A CN105859807A CN 105859807 A CN105859807 A CN 105859807A CN 201610225439 A CN201610225439 A CN 201610225439A CN 105859807 A CN105859807 A CN 105859807A
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- 239000000758 substrate Substances 0.000 title claims abstract description 32
- 239000003068 molecular probe Substances 0.000 title claims abstract description 13
- 108010022308 beta-1,4-galactosyltransferase I Proteins 0.000 title claims abstract description 9
- 238000002360 preparation method Methods 0.000 title claims abstract description 9
- 150000001875 compounds Chemical class 0.000 claims abstract description 25
- 238000006243 chemical reaction Methods 0.000 claims abstract description 13
- 238000000034 method Methods 0.000 claims abstract description 10
- -1 lactosyl group Chemical group 0.000 claims abstract description 9
- 125000002777 acetyl group Chemical group [H]C([H])([H])C(*)=O 0.000 claims abstract description 7
- 239000000126 substance Substances 0.000 claims abstract description 7
- NLFBCYMMUAKCPC-KQQUZDAGSA-N ethyl (e)-3-[3-amino-2-cyano-1-[(e)-3-ethoxy-3-oxoprop-1-enyl]sulfanyl-3-oxoprop-1-enyl]sulfanylprop-2-enoate Chemical compound CCOC(=O)\C=C\SC(=C(C#N)C(N)=O)S\C=C\C(=O)OCC NLFBCYMMUAKCPC-KQQUZDAGSA-N 0.000 claims description 10
- CJIJXIFQYOPWTF-UHFFFAOYSA-N 7-hydroxycoumarin Natural products O1C(=O)C=CC2=CC(O)=CC=C21 CJIJXIFQYOPWTF-UHFFFAOYSA-N 0.000 claims description 7
- ORHBXUUXSCNDEV-UHFFFAOYSA-N umbelliferone Chemical compound C1=CC(=O)OC2=CC(O)=CC=C21 ORHBXUUXSCNDEV-UHFFFAOYSA-N 0.000 claims description 7
- HFTAFOQKODTIJY-UHFFFAOYSA-N umbelliferone Natural products Cc1cc2C=CC(=O)Oc2cc1OCC=CC(C)(C)O HFTAFOQKODTIJY-UHFFFAOYSA-N 0.000 claims description 7
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 6
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 claims description 6
- WQDUMFSSJAZKTM-UHFFFAOYSA-N Sodium methoxide Chemical compound [Na+].[O-]C WQDUMFSSJAZKTM-UHFFFAOYSA-N 0.000 claims description 6
- VKYKSIONXSXAKP-UHFFFAOYSA-N hexamethylenetetramine Chemical compound C1N(C2)CN3CN1CN2C3 VKYKSIONXSXAKP-UHFFFAOYSA-N 0.000 claims description 4
- 229960000583 acetic acid Drugs 0.000 claims description 3
- MSWZFWKMSRAUBD-UHFFFAOYSA-N beta-D-galactosamine Natural products NC1C(O)OC(CO)C(O)C1O MSWZFWKMSRAUBD-UHFFFAOYSA-N 0.000 claims description 3
- 239000012362 glacial acetic acid Substances 0.000 claims description 3
- 229960002442 glucosamine Drugs 0.000 claims description 3
- 235000010299 hexamethylene tetramine Nutrition 0.000 claims description 3
- 229960004011 methenamine Drugs 0.000 claims description 3
- JRMUNVKIHCOMHV-UHFFFAOYSA-M tetrabutylammonium bromide Chemical compound [Br-].CCCC[N+](CCCC)(CCCC)CCCC JRMUNVKIHCOMHV-UHFFFAOYSA-M 0.000 claims description 3
- WIHIUTUAHOZVLE-UHFFFAOYSA-N 1,3-diethoxypropan-2-ol Chemical compound CCOCC(O)COCC WIHIUTUAHOZVLE-UHFFFAOYSA-N 0.000 claims description 2
- PSCXEUSWZWRCMQ-UHFFFAOYSA-N F[S](F)F Chemical compound F[S](F)F PSCXEUSWZWRCMQ-UHFFFAOYSA-N 0.000 claims description 2
- 230000006196 deacetylation Effects 0.000 claims description 2
- 238000003381 deacetylation reaction Methods 0.000 claims description 2
- 239000003444 phase transfer catalyst Substances 0.000 claims description 2
- 239000002994 raw material Substances 0.000 claims description 2
- 238000006467 substitution reaction Methods 0.000 claims description 2
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- 230000000694 effects Effects 0.000 abstract description 6
- 230000007062 hydrolysis Effects 0.000 abstract description 4
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- 125000002519 galactosyl group Chemical group C1([C@H](O)[C@@H](O)[C@@H](O)[C@H](O1)CO)* 0.000 abstract description 3
- 230000000269 nucleophilic effect Effects 0.000 abstract description 3
- 230000006098 transglycosylation Effects 0.000 abstract description 3
- 238000005918 transglycosylation reaction Methods 0.000 abstract description 3
- HSCJRCZFDFQWRP-ABVWGUQPSA-N UDP-alpha-D-galactose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1OP(O)(=O)OP(O)(=O)OC[C@@H]1[C@@H](O)[C@@H](O)[C@H](N2C(NC(=O)C=C2)=O)O1 HSCJRCZFDFQWRP-ABVWGUQPSA-N 0.000 abstract description 2
- HSCJRCZFDFQWRP-UHFFFAOYSA-N Uridindiphosphoglukose Natural products OC1C(O)C(O)C(CO)OC1OP(O)(=O)OP(O)(=O)OCC1C(O)C(O)C(N2C(NC(=O)C=C2)=O)O1 HSCJRCZFDFQWRP-UHFFFAOYSA-N 0.000 abstract description 2
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- 238000003379 elimination reaction Methods 0.000 abstract 1
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 9
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- YMWUJEATGCHHMB-UHFFFAOYSA-N methylene chloride Substances ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 6
- 239000007787 solid Substances 0.000 description 6
- HSHNITRMYYLLCV-UHFFFAOYSA-N beta-methyl umbelliferone Natural products C1=C(O)C=CC2=C1OC(=O)C=C2C HSHNITRMYYLLCV-UHFFFAOYSA-N 0.000 description 5
- 239000000523 sample Substances 0.000 description 5
- 150000008131 glucosides Chemical group 0.000 description 4
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- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 3
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 239000000706 filtrate Substances 0.000 description 3
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 3
- CSNNHWWHGAXBCP-UHFFFAOYSA-L magnesium sulphate Substances [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 239000012074 organic phase Substances 0.000 description 3
- AZQWKYJCGOJGHM-UHFFFAOYSA-N para-benzoquinone Natural products O=C1C=CC(=O)C=C1 AZQWKYJCGOJGHM-UHFFFAOYSA-N 0.000 description 3
- 238000003786 synthesis reaction Methods 0.000 description 3
- FKLJPTJMIBLJAV-UHFFFAOYSA-N Compound IV Chemical compound O1N=C(C)C=C1CCCCCCCOC1=CC=C(C=2OCCN=2)C=C1 FKLJPTJMIBLJAV-UHFFFAOYSA-N 0.000 description 2
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 2
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 2
- 206010027476 Metastases Diseases 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 238000006555 catalytic reaction Methods 0.000 description 2
- ZYGHJZDHTFUPRJ-UHFFFAOYSA-N coumarin Chemical compound C1=CC=C2OC(=O)C=CC2=C1 ZYGHJZDHTFUPRJ-UHFFFAOYSA-N 0.000 description 2
- 238000013461 design Methods 0.000 description 2
- 238000009792 diffusion process Methods 0.000 description 2
- 229930182470 glycoside Natural products 0.000 description 2
- 239000008101 lactose Substances 0.000 description 2
- 230000009401 metastasis Effects 0.000 description 2
- 229950006780 n-acetylglucosamine Drugs 0.000 description 2
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- 235000002639 sodium chloride Nutrition 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 239000012265 solid product Substances 0.000 description 2
- NWUYHJFMYQTDRP-UHFFFAOYSA-N 1,2-bis(ethenyl)benzene;1-ethenyl-2-ethylbenzene;styrene Chemical compound C=CC1=CC=CC=C1.CCC1=CC=CC=C1C=C.C=CC1=CC=CC=C1C=C NWUYHJFMYQTDRP-UHFFFAOYSA-N 0.000 description 1
- 102100027321 Beta-1,4-galactosyltransferase 7 Human genes 0.000 description 1
- 206010006187 Breast cancer Diseases 0.000 description 1
- 208000026310 Breast neoplasm Diseases 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 108700023372 Glycosyltransferases Proteins 0.000 description 1
- 102000051366 Glycosyltransferases Human genes 0.000 description 1
- NNJVILVZKWQKPM-UHFFFAOYSA-N Lidocaine Chemical compound CCN(CC)CC(=O)NC1=C(C)C=CC=C1C NNJVILVZKWQKPM-UHFFFAOYSA-N 0.000 description 1
- 206010058467 Lung neoplasm malignant Diseases 0.000 description 1
- OVRNDRQMDRJTHS-UHFFFAOYSA-N N-acelyl-D-glucosamine Natural products CC(=O)NC1C(O)OC(CO)C(O)C1O OVRNDRQMDRJTHS-UHFFFAOYSA-N 0.000 description 1
- OVRNDRQMDRJTHS-FMDGEEDCSA-N N-acetyl-beta-D-glucosamine Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O OVRNDRQMDRJTHS-FMDGEEDCSA-N 0.000 description 1
- 108091034117 Oligonucleotide Proteins 0.000 description 1
- 206010060862 Prostate cancer Diseases 0.000 description 1
- 208000000236 Prostatic Neoplasms Diseases 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 108010063641 Xylosylprotein 4-beta-galactosyltransferase Proteins 0.000 description 1
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- QZHPTGXQGDFGEN-UHFFFAOYSA-N chromene Chemical compound C1=CC=C2C=C[CH]OC2=C1 QZHPTGXQGDFGEN-UHFFFAOYSA-N 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 229960000956 coumarin Drugs 0.000 description 1
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- 239000012847 fine chemical Substances 0.000 description 1
- 238000000799 fluorescence microscopy Methods 0.000 description 1
- 229930182478 glucoside Natural products 0.000 description 1
- 150000002338 glycosides Chemical class 0.000 description 1
- LIIALPBMIOVAHH-UHFFFAOYSA-N herniarin Chemical class C1=CC(=O)OC2=CC(OC)=CC=C21 LIIALPBMIOVAHH-UHFFFAOYSA-N 0.000 description 1
- 239000004312 hexamethylene tetramine Substances 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 230000016507 interphase Effects 0.000 description 1
- 239000003456 ion exchange resin Substances 0.000 description 1
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- 239000011159 matrix material Substances 0.000 description 1
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- 229910052757 nitrogen Inorganic materials 0.000 description 1
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- 238000012216 screening Methods 0.000 description 1
- SFZCNBIFKDRMGX-UHFFFAOYSA-N sulfur hexafluoride Chemical compound FS(F)(F)(F)(F)F SFZCNBIFKDRMGX-UHFFFAOYSA-N 0.000 description 1
- 229960000909 sulfur hexafluoride Drugs 0.000 description 1
- 208000010570 urinary bladder carcinoma Diseases 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
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Classifications
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H17/00—Compounds containing heterocyclic radicals directly attached to hetero atoms of saccharide radicals
- C07H17/04—Heterocyclic radicals containing only oxygen as ring hetero atoms
- C07H17/06—Benzopyran radicals
- C07H17/065—Benzo[b]pyrans
- C07H17/075—Benzo[b]pyran-2-ones
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/001—Preparation for luminescence or biological staining
- A61K49/0013—Luminescence
- A61K49/0017—Fluorescence in vivo
- A61K49/0019—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules
- A61K49/0021—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules the fluorescent group being a small organic molecule
- A61K49/0039—Coumarin dyes
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H1/00—Processes for the preparation of sugar derivatives
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- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
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- C09K11/00—Luminescent, e.g. electroluminescent, chemiluminescent materials
- C09K11/06—Luminescent, e.g. electroluminescent, chemiluminescent materials containing organic luminescent materials
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6486—Measuring fluorescence of biological material, e.g. DNA, RNA, cells
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- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K2211/00—Chemical nature of organic luminescent or tenebrescent compounds
- C09K2211/10—Non-macromolecular compounds
- C09K2211/1018—Heterocyclic compounds
- C09K2211/1025—Heterocyclic compounds characterised by ligands
- C09K2211/1088—Heterocyclic compounds characterised by ligands containing oxygen as the only heteroatom
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Abstract
The invention relates to a chemically synthesized beta-1,4-galactosyltransferase I (beta-1,4-GalT I) substrate fluorescent molecular probe targeting to tumor cells, and an intermediate and a preparation method thereof. The chemical structure of the beta-1,4-GalT I substrate fluorescent molecular probe is shown in the description, and Ac in the formula represents an acetyl group. Tumor cell strains are screened to obtain high-expression beta-1,4-GalT I adopted as a target, a 8-difluoromethyl-7-hydroxycoumarin-beta-acetylglucoside specific substrate is designed and synthesized based on an ASPs strategy, the galactosyl group of a donor UDP-Gal is transferred to the substrate through using the transglycosylation of beta-1,4-GalT I in order to generate a lactosyl group, aglucones are released in the glycosidase hydrolysis process and undergo an elimination reaction to form a quinonylmethyl compound with high reaction activity, and the quinonylmethyl compound is covalently combined with the nucleophilic substrate nearby the enzyme to finally complete targeting fluorescent labeling process of the tumor cells, so early diagnosis of the tumors is realized.
Description
Technical field
The present invention relates to a kind of chemical synthesis preparation can the β-1,4-Galactosyltransferase I of targets neoplastic cells
Substrate fluorescence molecular probe and intermediate product and preparation method thereof, the invention belongs to field of fine chemical.
Background technology
It is handmarking based on bioactive probe molecule technology (Activity-Based Probes, ABPs)
Enzyme specificity substrate molecule and targeted cells (enzyme) combine and generate compound, thus realize enzyme, cell
Targeting selectivity marks.Visited by the fluorescence molecule of screening tumor cell specific zymolyte based on ABPs strategy
Pin realizes the targeting fluorescence imaging of tumour, has that targeting is strong and the advantage such as radiationless exposure.
Cell surface β-1,4-Galactosyltransferase I (β-1,4-GalT I) have with fertilization, neuron thin
The important biomolecule merit that the processes such as born of the same parents' migration, cancer metastasis, epithelial cell proliferation and cellular signal transduction are relevant
Energy.β-the 1,4-GalT I of cell surface with the combination of the N-Acetyl-D-glucosamine in cytoplasm be cell and
The main cause of matter adhesion, it participates in iuntercellular, the characteristic of cell-matrix interphase interaction determines it swollen
Tumor metastasis has important effect.Research shows when cell and laminin occur to adhere to, cell surface
β-1,4-GalT I level raise 300%.Therefore β-1,4-GalT I specific substrate probe molecule targeting is designed
Tumour cell realizes tumour diagnosis in early days and treatment is possibly realized.
Summary of the invention
For above-mentioned technical problem, the present invention provides a kind of β-Isosorbide-5-Nitrae-Galactosyltransferase I Substrate fluorescence molecule
Probe and intermediate product and preparation method thereof.
Concrete technical scheme is:
β-Isosorbide-5-Nitrae-Galactosyltransferase I Substrate fluorescence molecular probe, for compound V, chemical constitution is:
Wherein, Ac represents acetyl group.
Prepare the midbody compound III of β-Isosorbide-5-Nitrae-Galactosyltransferase I Substrate fluorescence molecular probe, chemistry knot
Structure is:
Wherein, Ac represents acetyl group.
Prepare the midbody compound IV of β-Isosorbide-5-Nitrae-Galactosyltransferase I Substrate fluorescence molecular probe, chemistry knot
Structure is:
Wherein, Ac represents acetyl group.
Midbody compound III, compounds Ⅳ, compound V preparation method:
With chemical compounds I i.e. umbelliferone as raw material, introduce formaldehyde in C-8 position with glacial acetic acid and methenamine
Obtain compound ii；It is the bar of phase transfer catalyst with full acetylated bromo Glucosamine at TBAB
Carry out substitution reaction under part, obtain compound III；Compound III and diethylin sulfur trifluoride carry out fluoro-reaction
Prepare compounds Ⅳ；Target compound V is prepared through sodium methoxide solution deacetylation after.
Umbelliferone, also known as umbelliferone, is a kind of hyperfluorescence dyestuff, using umbelliferone labeled substrate as molecule
Probe, under enzyme catalysis, product generation intramolecular β-elimination is reacted and is made the umbelliferone on substrate come off, logical
The change reflection enzyme crossing fluorescence intensity is lived.Utilize the umbelliferone sugar that glycosidase activity is marked in recent years
Glycosides substrate mainly has two classes.1. 4-methyl umbelliferone β-D-glucopyranoside substrate, first by glycosyl transferase
Catalysis generates corresponding oligonucleotide chain, and recycling glycosidase hydrolysis discharges fluorescence activity material 4-methyl umbelliferone,
Fluorophor owing to generating is fixed without corresponding chemical bond with between the enzyme marked or cell, makes fluorescence
Group causes marking inconspicuous the most failed in each intercellular diffusion.2. 8-halogenated methyl-umbelliferone β
-pyranoside, Withers etc. utilizes such substrate to sub-elect 1～1 from the yeast cells that a large amount of glycosidases inactivate
0% enzymatic activity yeast, the glucoside unit that fluoroglycoside substrate discharges with glycosidase hydrolysis generates higher through eliminating reaction
The quinone methyl compound of reactivity, nucleophilicity component quickly and near enzyme or cellular component are with stable covalency
Bond is closed, thus is prevented effectively from the diffusion of fluorescent material.
The parent nucleus chromene of coumarin kind compound is the colourless substance not having fluorescence, but the derivative after replacing,
After introducing electron donating group on particularly C-7 position, form push and pull component, just obtain hyperfluorescence material.This
The selectivity substrate 8-difluoromethyl-umbelliferone-β-acetylamino of invention design synthesis β-1,4-GalT I
Glucoside (compound V) selectivity substrate, by β-Isosorbide-5-Nitrae-GalT I by transglycosylation by donor UD
The galactosyl of P-Gal is transferred to substrate and generates lactose base, and glycosidase hydrolysis discharges glucoside unit, eliminates reaction shape
Become the quinone methyl compound of high reaction activity, with the nucleophilic substrate covalent bond near enzyme, it is achieved to tumour cell
Targeting fluorescence labeling, following process:
β-1,4-Galactosyltransferase I Substrate fluorescence molecular probe that the present invention provides and intermediate product and system thereof
Preparation Method, screens high table in the tumor cell lines such as China's lung cancer occurred frequently, breast cancer, prostate cancer, carcinoma of urinary bladder
The β reached-Isosorbide-5-Nitrae Galactosyltransferase I is target spot, based on ASPs strategy design synthesis 8-difluoromethyl-7-hydroxyl
Butylcoumariii-β-acetylglucosamine glycosides (compound V) selectivity substrate, is first led to by β-Isosorbide-5-Nitrae-GalT I
Crossing transglycosylation and the galactosyl of donor UDP-Gal is transferred to substrate generation lactose base, glycosidase hydrolyzes
Discharge glucoside unit, eliminate reaction and form the quinone methyl compound of high reaction activity, be total to the nucleophilic substrate near enzyme
Valency combines, and is finally completed the targeting fluorescence labeling process of tumour cell, thus realizes the early diagnosis of tumour.
Detailed description of the invention
It is described in conjunction with the embodiments the detailed description of the invention of the present invention.
1, the building-up process of compound ii
Weigh 10.0g 4-methyl umbelliferone and 20.0g hexamethylenetetramine in three-neck flask, add glacial acetic acid；
95 DEG C of stirring reaction 5.5h, add dilute HCl in reactant liquor, continue reaction 30min；Reactant liquor is cooled to room
Temperature, adds water；Absolute ether extracts, saturated aqueous common salt washing organic phase；Anhydrous MgSO4It is dried, suction filtration；
Reduced pressure concentration filtrate prepares solid product, and absolute ethyl alcohol recrystallizes to obtain yellow solid product (compound ii) 3.7g.
2, the building-up process of compound III
Weigh 11.5g compound ii, full acetylated bromo Glucosamine and TBAB, join circle
In end flask, CH2Cl2After dissolving, it is slowly added to NaOH solution (1.0mol/L, pH=11), stirring
React 12 hours；Being extracted with ethyl acetate, wash organic phase, saturated common salt is washed, anhydrous MgSO4It is dried,
Suction filtration；Filtrate is concentrated in vacuo to thick solid, re-crystallizing in ethyl acetate, is vacuum dried to obtain white solid (compound
Ⅲ)18.1g。
3, the building-up process of compounds Ⅳ
Weighing 8.7g compound III in 250mL round-bottomed flask, dichloromethane dissolves, and adds lignocaine three
Sulfur fluoride (DAST), nitrogen is protected；Stirring reaction 3.5 hours, adds appropriate ice and stops reaction；Dichloromethane
Alkane extracts, and washes organic phase, and saturated aqueous common salt washs；Anhydrous MgSO4It is dried, suction filtration；Filter is concentrated in vacuo
Liquid, to thick solid, re-crystallizing in ethyl acetate, is vacuum dried to obtain white solid (compounds Ⅳ) 7.51g.
4, the building-up process of compound V
Weigh Compound IV 3.0g is in round-bottomed flask, after methyl alcohol stirring and dissolving, adds sodium methoxide solution, nitrogen
Gas shielded；Reaction 40min is stirred at room temperature, adds appropriate 120H ion exchange resin and neutralize；Suction filtration, concentrates
Filtrate is to thick solid；Recrystallize with the mixed liquor of methyl alcohol and absolute ether, obtain white solid 0.9g, i.e. chemical combination
Thing V.
Claims (4)
1. β-Isosorbide-5-Nitrae-Galactosyltransferase I Substrate fluorescence molecular probe, for compound V, is characterised by, changes
Structure is:
Wherein, Ac represents acetyl group.
β-1,4-Galactosyltransferase I Substrate fluorescence molecular probe the most according to claim 1 is in the middle of it
Body compound III, is characterised by, chemical constitution is:
Wherein, Ac represents acetyl group.
β-1,4-Galactosyltransferase I Substrate fluorescence molecular probe the most according to claim 1 is in the middle of it
Body compounds Ⅳ, it is characterised in that chemical constitution is:
Wherein, Ac represents acetyl group.
The preparation of β-1,4-Galactosyltransferase I Substrate fluorescence molecular probe the most according to claim 1
Method:
With chemical compounds I i.e. umbelliferone as raw material, introduce formaldehyde in C-8 position with glacial acetic acid and methenamine
Obtain compound ii；It is the bar of phase transfer catalyst with full acetylated bromo Glucosamine at TBAB
Carry out substitution reaction under part, obtain compound III；Compound III and diethylin sulfur trifluoride carry out fluoro-reaction
Prepare compounds Ⅳ；Target compound V is prepared through sodium methoxide solution deacetylation after.
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| CN109824743A (en) * | 2019-03-04 | 2019-05-31 | 大连医科大学 | It is a kind of detect N-acetyl-β-D-glucosaminidase fluorescence probe and its application |
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| CN102041295A (en) * | 2009-10-19 | 2011-05-04 | 复旦大学 | Application of beta-1,4 galactosy transferase I in preparing liver cancer diagnostic preparation |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109824743A (en) * | 2019-03-04 | 2019-05-31 | 大连医科大学 | It is a kind of detect N-acetyl-β-D-glucosaminidase fluorescence probe and its application |
| CN109824743B (en) * | 2019-03-04 | 2021-11-23 | 大连医科大学 | Fluorescent probe for detecting N-acetyl-beta-D-glucosaminidase and application thereof |
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