CN114736264A - Tau蛋白可视化PROTAC降解化合物及其制备方法和应用 - Google Patents
Tau蛋白可视化PROTAC降解化合物及其制备方法和应用 Download PDFInfo
- Publication number
- CN114736264A CN114736264A CN202210387786.1A CN202210387786A CN114736264A CN 114736264 A CN114736264 A CN 114736264A CN 202210387786 A CN202210387786 A CN 202210387786A CN 114736264 A CN114736264 A CN 114736264A
- Authority
- CN
- China
- Prior art keywords
- stirring
- tau protein
- drying
- room temperature
- 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.)
- Granted
Links
- 102000013498 tau Proteins Human genes 0.000 title claims abstract description 86
- 108010026424 tau Proteins Proteins 0.000 title claims abstract description 86
- 150000001875 compounds Chemical class 0.000 title claims abstract description 85
- 238000006731 degradation reaction Methods 0.000 title claims abstract description 62
- 230000015556 catabolic process Effects 0.000 title claims abstract description 55
- 108010026668 snake venom protein C activator Proteins 0.000 title claims abstract description 29
- 238000011865 proteolysis targeting chimera technique Methods 0.000 title claims abstract description 28
- 229940124823 proteolysis targeting chimeric molecule Drugs 0.000 title claims abstract description 28
- 238000002360 preparation method Methods 0.000 title claims abstract description 16
- 230000000007 visual effect Effects 0.000 title abstract description 10
- 239000003446 ligand Substances 0.000 claims abstract description 44
- 102000006275 Ubiquitin-Protein Ligases Human genes 0.000 claims abstract description 18
- 108010083111 Ubiquitin-Protein Ligases Proteins 0.000 claims abstract description 18
- 230000017854 proteolysis Effects 0.000 claims abstract description 11
- 201000010099 disease Diseases 0.000 claims abstract description 10
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims abstract description 10
- 239000000523 sample Substances 0.000 claims abstract description 8
- 230000000593 degrading effect Effects 0.000 claims abstract description 7
- 238000003756 stirring Methods 0.000 claims description 97
- 238000006243 chemical reaction Methods 0.000 claims description 84
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 78
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 78
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 claims description 48
- JGFZNNIVVJXRND-UHFFFAOYSA-N N,N-Diisopropylethylamine (DIPEA) Chemical compound CCN(C(C)C)C(C)C JGFZNNIVVJXRND-UHFFFAOYSA-N 0.000 claims description 44
- 238000001035 drying Methods 0.000 claims description 44
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 35
- VHYFNPMBLIVWCW-UHFFFAOYSA-N 4-Dimethylaminopyridine Chemical compound CN(C)C1=CC=NC=C1 VHYFNPMBLIVWCW-UHFFFAOYSA-N 0.000 claims description 32
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 29
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 27
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 25
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 claims description 24
- BWHMMNNQKKPAPP-UHFFFAOYSA-L potassium carbonate Chemical compound [K+].[K+].[O-]C([O-])=O BWHMMNNQKKPAPP-UHFFFAOYSA-L 0.000 claims description 24
- 238000010438 heat treatment Methods 0.000 claims description 20
- 238000000034 method Methods 0.000 claims description 20
- 239000007787 solid Substances 0.000 claims description 19
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 claims description 18
- 238000001816 cooling Methods 0.000 claims description 16
- 229960000549 4-dimethylaminophenol Drugs 0.000 claims description 15
- 239000007821 HATU Substances 0.000 claims description 15
- 230000015572 biosynthetic process Effects 0.000 claims description 15
- 239000012043 crude product Substances 0.000 claims description 15
- 238000004440 column chromatography Methods 0.000 claims description 14
- 238000001914 filtration Methods 0.000 claims description 14
- 239000000543 intermediate Substances 0.000 claims description 13
- 238000003786 synthesis reaction Methods 0.000 claims description 13
- 208000024827 Alzheimer disease Diseases 0.000 claims description 12
- XHXFXVLFKHQFAL-UHFFFAOYSA-N phosphoryl trichloride Chemical compound ClP(Cl)(Cl)=O XHXFXVLFKHQFAL-UHFFFAOYSA-N 0.000 claims description 12
- UVSMNLNDYGZFPF-UHFFFAOYSA-N pomalidomide Chemical compound O=C1C=2C(N)=CC=CC=2C(=O)N1C1CCC(=O)NC1=O UVSMNLNDYGZFPF-UHFFFAOYSA-N 0.000 claims description 12
- 229960000688 pomalidomide Drugs 0.000 claims description 12
- 229910000027 potassium carbonate Inorganic materials 0.000 claims description 12
- 239000012295 chemical reaction liquid Substances 0.000 claims description 10
- 238000010898 silica gel chromatography Methods 0.000 claims description 10
- 239000003795 chemical substances by application Substances 0.000 claims description 9
- 230000008569 process Effects 0.000 claims description 9
- 230000002829 reductive effect Effects 0.000 claims description 9
- WQDUMFSSJAZKTM-UHFFFAOYSA-N Sodium methoxide Chemical compound [Na+].[O-]C WQDUMFSSJAZKTM-UHFFFAOYSA-N 0.000 claims description 7
- 239000005457 ice water Substances 0.000 claims description 7
- MLIREBYILWEBDM-UHFFFAOYSA-N cyanoacetic acid Chemical compound OC(=O)CC#N MLIREBYILWEBDM-UHFFFAOYSA-N 0.000 claims description 6
- VAYGXNSJCAHWJZ-UHFFFAOYSA-N dimethyl sulfate Chemical compound COS(=O)(=O)OC VAYGXNSJCAHWJZ-UHFFFAOYSA-N 0.000 claims description 6
- CMSYDJVRTHCWFP-UHFFFAOYSA-N triphenylphosphane;hydrobromide Chemical compound Br.C1=CC=CC=C1P(C=1C=CC=CC=1)C1=CC=CC=C1 CMSYDJVRTHCWFP-UHFFFAOYSA-N 0.000 claims description 5
- 238000005406 washing Methods 0.000 claims description 5
- 238000009825 accumulation Methods 0.000 claims description 4
- 238000000605 extraction Methods 0.000 claims description 4
- 238000000926 separation method Methods 0.000 claims description 4
- FRHRVQQUICVJDG-UHFFFAOYSA-M 1,3-dioxolan-2-ylmethyl(triphenyl)phosphanium;bromide Chemical compound [Br-].C=1C=CC=CC=1[P+](C=1C=CC=CC=1)(C=1C=CC=CC=1)CC1OCCO1 FRHRVQQUICVJDG-UHFFFAOYSA-M 0.000 claims description 3
- CRAUTELYXAAAPW-UHFFFAOYSA-N 2-(2,6-dioxopiperidin-3-yl)-4-fluoroisoindole-1,3-dione Chemical compound O=C1C=2C(F)=CC=CC=2C(=O)N1C1CCC(=O)NC1=O CRAUTELYXAAAPW-UHFFFAOYSA-N 0.000 claims description 3
- WNRGWPVJGDABME-UHFFFAOYSA-N 3,5-Dimethoxyaniline Chemical compound COC1=CC(N)=CC(OC)=C1 WNRGWPVJGDABME-UHFFFAOYSA-N 0.000 claims description 3
- 229910019213 POCl3 Inorganic materials 0.000 claims description 3
- KWYUFKZDYYNOTN-UHFFFAOYSA-M Potassium hydroxide Chemical compound [OH-].[K+] KWYUFKZDYYNOTN-UHFFFAOYSA-M 0.000 claims description 3
- HJMZMZRCABDKKV-UHFFFAOYSA-N carbonocyanidic acid Chemical compound OC(=O)C#N HJMZMZRCABDKKV-UHFFFAOYSA-N 0.000 claims description 3
- 229960004942 lenalidomide Drugs 0.000 claims description 3
- GOTYRUGSSMKFNF-UHFFFAOYSA-N lenalidomide Chemical compound C1C=2C(N)=CC=CC=2C(=O)N1C1CCC(=O)NC1=O GOTYRUGSSMKFNF-UHFFFAOYSA-N 0.000 claims description 3
- 230000007935 neutral effect Effects 0.000 claims description 3
- 239000002994 raw material Substances 0.000 claims description 3
- XSDFQFQJASDMMB-UHFFFAOYSA-N tert-butyl n-(1-aminopentyl)carbamate Chemical compound CCCCC(N)NC(=O)OC(C)(C)C XSDFQFQJASDMMB-UHFFFAOYSA-N 0.000 claims description 3
- 238000012544 monitoring process Methods 0.000 claims description 2
- 238000010992 reflux Methods 0.000 claims description 2
- 230000000694 effects Effects 0.000 abstract description 28
- 238000013461 design Methods 0.000 abstract description 7
- 150000003384 small molecules Chemical class 0.000 abstract description 5
- 230000009467 reduction Effects 0.000 abstract description 4
- 238000001514 detection method Methods 0.000 abstract description 2
- 210000004027 cell Anatomy 0.000 description 25
- 238000005160 1H NMR spectroscopy Methods 0.000 description 16
- HEDRZPFGACZZDS-MICDWDOJSA-N Trichloro(2H)methane Chemical compound [2H]C(Cl)(Cl)Cl HEDRZPFGACZZDS-MICDWDOJSA-N 0.000 description 14
- 238000001644 13C nuclear magnetic resonance spectroscopy Methods 0.000 description 12
- 239000012071 phase Substances 0.000 description 11
- 239000000203 mixture Substances 0.000 description 10
- -1 small molecule compounds Chemical class 0.000 description 10
- 230000008685 targeting Effects 0.000 description 10
- 102000004169 proteins and genes Human genes 0.000 description 9
- 108090000623 proteins and genes Proteins 0.000 description 9
- IAZDPXIOMUYVGZ-WFGJKAKNSA-N Dimethyl sulfoxide Chemical compound [2H]C([2H])([2H])S(=O)C([2H])([2H])[2H] IAZDPXIOMUYVGZ-WFGJKAKNSA-N 0.000 description 8
- 230000008045 co-localization Effects 0.000 description 7
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 6
- 238000004458 analytical method Methods 0.000 description 6
- 238000009833 condensation Methods 0.000 description 6
- 230000005494 condensation Effects 0.000 description 6
- 238000000746 purification Methods 0.000 description 6
- 230000006870 function Effects 0.000 description 5
- 230000003834 intracellular effect Effects 0.000 description 5
- 238000001543 one-way ANOVA Methods 0.000 description 5
- 150000001732 carboxylic acid derivatives Chemical class 0.000 description 4
- 230000008859 change Effects 0.000 description 4
- 239000007795 chemical reaction product Substances 0.000 description 4
- 239000003814 drug Substances 0.000 description 4
- 239000011541 reaction mixture Substances 0.000 description 4
- 101000746263 Conus leopardus Conotoxin Lp5.1 Proteins 0.000 description 3
- 201000011240 Frontotemporal dementia Diseases 0.000 description 3
- 238000011156 evaluation Methods 0.000 description 3
- 239000007850 fluorescent dye Substances 0.000 description 3
- 238000003384 imaging method Methods 0.000 description 3
- 210000002682 neurofibrillary tangle Anatomy 0.000 description 3
- 210000002569 neuron Anatomy 0.000 description 3
- 230000001575 pathological effect Effects 0.000 description 3
- 239000003208 petroleum Substances 0.000 description 3
- 238000002600 positron emission tomography Methods 0.000 description 3
- 238000009987 spinning Methods 0.000 description 3
- OBMZMSLWNNWEJA-XNCRXQDQSA-N C1=CC=2C(C[C@@H]3NC(=O)[C@@H](NC(=O)[C@H](NC(=O)N(CC#CCN(CCCC[C@H](NC(=O)[C@@H](CC4=CC=CC=C4)NC3=O)C(=O)N)CC=C)NC(=O)[C@@H](N)C)CC3=CNC4=C3C=CC=C4)C)=CNC=2C=C1 Chemical compound C1=CC=2C(C[C@@H]3NC(=O)[C@@H](NC(=O)[C@H](NC(=O)N(CC#CCN(CCCC[C@H](NC(=O)[C@@H](CC4=CC=CC=C4)NC3=O)C(=O)N)CC=C)NC(=O)[C@@H](N)C)CC3=CNC4=C3C=CC=C4)C)=CNC=2C=C1 OBMZMSLWNNWEJA-XNCRXQDQSA-N 0.000 description 2
- OKKJLVBELUTLKV-MZCSYVLQSA-N Deuterated methanol Chemical compound [2H]OC([2H])([2H])[2H] OKKJLVBELUTLKV-MZCSYVLQSA-N 0.000 description 2
- 101710176384 Peptide 1 Proteins 0.000 description 2
- 102000004245 Proteasome Endopeptidase Complex Human genes 0.000 description 2
- 108090000708 Proteasome Endopeptidase Complex Proteins 0.000 description 2
- 230000002159 abnormal effect Effects 0.000 description 2
- 238000003125 immunofluorescent labeling Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 229920000642 polymer Polymers 0.000 description 2
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 2
- 108090000765 processed proteins & peptides Proteins 0.000 description 2
- 230000001105 regulatory effect Effects 0.000 description 2
- 230000000717 retained effect Effects 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 230000034512 ubiquitination Effects 0.000 description 2
- 238000010798 ubiquitination Methods 0.000 description 2
- 238000010200 validation analysis Methods 0.000 description 2
- 230000006269 (delayed) early viral mRNA transcription Effects 0.000 description 1
- 208000037259 Amyloid Plaque Diseases 0.000 description 1
- 102000010909 Monoamine Oxidase Human genes 0.000 description 1
- 108010062431 Monoamine oxidase Proteins 0.000 description 1
- 241000699666 Mus <mouse, genus> Species 0.000 description 1
- 229940127576 PROTAC degrader Drugs 0.000 description 1
- 108010033276 Peptide Fragments Proteins 0.000 description 1
- 102000007079 Peptide Fragments Human genes 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- 102000044159 Ubiquitin Human genes 0.000 description 1
- 108090000848 Ubiquitin Proteins 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000002776 aggregation Effects 0.000 description 1
- 238000004220 aggregation Methods 0.000 description 1
- 150000001335 aliphatic alkanes Chemical group 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 239000008346 aqueous phase Substances 0.000 description 1
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 208000010877 cognitive disease Diseases 0.000 description 1
- 238000004891 communication Methods 0.000 description 1
- 238000012790 confirmation Methods 0.000 description 1
- 210000004292 cytoskeleton Anatomy 0.000 description 1
- 230000003413 degradative effect Effects 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 238000001647 drug administration Methods 0.000 description 1
- 230000001094 effect on targets Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 230000009088 enzymatic function Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000000684 flow cytometry Methods 0.000 description 1
- 238000001506 fluorescence spectroscopy Methods 0.000 description 1
- 238000001215 fluorescent labelling Methods 0.000 description 1
- 230000002452 interceptive effect Effects 0.000 description 1
- 230000000968 intestinal effect Effects 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 230000001483 mobilizing effect Effects 0.000 description 1
- 239000000178 monomer Substances 0.000 description 1
- 210000003061 neural cell Anatomy 0.000 description 1
- 208000015122 neurodegenerative disease Diseases 0.000 description 1
- 230000036961 partial effect Effects 0.000 description 1
- 230000007119 pathological manifestation Effects 0.000 description 1
- 230000000750 progressive effect Effects 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 230000004845 protein aggregation Effects 0.000 description 1
- 230000018883 protein targeting Effects 0.000 description 1
- 230000007115 recruitment Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 239000000741 silica gel Substances 0.000 description 1
- 229910002027 silica gel Inorganic materials 0.000 description 1
- SUBJHSREKVAVAR-UHFFFAOYSA-N sodium;methanol;methanolate Chemical compound [Na+].OC.[O-]C SUBJHSREKVAVAR-UHFFFAOYSA-N 0.000 description 1
- 238000007619 statistical method Methods 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 238000001308 synthesis method Methods 0.000 description 1
- 230000002123 temporal effect Effects 0.000 description 1
- 230000036962 time dependent Effects 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 238000001262 western blot Methods 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
- C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
- C07K5/06—Dipeptides
- C07K5/06008—Dipeptides with the first amino acid being neutral
- C07K5/06017—Dipeptides with the first amino acid being neutral and aliphatic
- C07K5/06034—Dipeptides with the first amino acid being neutral and aliphatic the side chain containing 2 to 4 carbon atoms
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
- A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
- C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
- C07D401/04—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings directly linked by a ring-member-to-ring-member bond
-
- 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
- C09K11/00—Luminescent, e.g. electroluminescent, chemiluminescent materials
- C09K11/06—Luminescent, e.g. electroluminescent, chemiluminescent materials containing organic luminescent materials
-
- 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
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
-
- 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
- C09K2211/1003—Carbocyclic compounds
- C09K2211/1007—Non-condensed systems
-
- 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
- C09K2211/1029—Heterocyclic compounds characterised by ligands containing one nitrogen atom as the heteroatom
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups
Abstract
本发明提供一类Tau蛋白可视化PROTAC降解化合物及其制备方法和应用,该类化合物是将E3连接酶配体先与linker缩合制备出中间体,再与靶头探针分子相连后得到的,在靶头选取上不仅保留靶头小分子与Tau蛋白的选择性结合作用,还赋予了其产生近红外荧光的全新功能,以使得目标化合物在与Tau蛋白结合时,除了发挥靶向降解Tau蛋白的功能,还能发出可检测的近红外荧光,从而使Tau蛋白的降解过程可视化;D‑12与D‑16均表现出了与p‑Tau减少相似的荧光趋势,实现了可视化的设计,有望对蛋白降解进程进行更为直观的观察与调控,为疾病检测手段与治疗手段相结合提供可能。
Description
技术领域
本发明属于医药合成化工技术领域,具体涉及Tau蛋白可视化PROTAC降解化合物及其制备方法和应用。
背景技术
阿尔兹海默症(Alzheimer’s disease,AD)是一种起病隐匿、进行性发展的神经系统退行疾病,主要表现为认知功能障碍,已经成为了全球致死率增长最快的疾病之一。然而目前临床批准的药物大多只能够延缓疾病的进程、减轻症状,并不能有效治愈AD。对于AD的病因假说主要有淀粉样斑块、神经元纤维缠结和肠道微生物等,其中过度磷酸化的Tau蛋白聚集成为不溶性神经元纤维缠结,对神经细胞内外的生化通讯产生破坏,同时也会对细胞骨架有所损伤。
蛋白降解靶向嵌合体(Proteolysis Targeting Chimeras,PROTAC)是一项可以通过泛素-蛋白酶体系统选择性降解细胞内蛋白的新技术,它更贴近于临床治疗,可以应用于各种与蛋白相关的疾病,PROTACs分子由三部分组成,分别是靶向蛋白的结合体、E3连接酶招募部分和两者之间的一段linker。PROTACs可以操纵细胞内固有的泛素蛋白酶体系统(UPS)来发挥其降解功能,减少目标蛋白。因此,通过构建靶向磷酸化Tau蛋白的可视化PROTACs,有望更直接造成p-Tau减少的生理结果,从而缓解甚至阻断神经纤维缠结的形成,减轻病理表现。
神经细胞内Tau蛋白的异常聚集是阿尔兹海默症患者一个突出的生理现象。Tau蛋白的高水平表达是促进其异常聚集的原因之一,同时也会间接调控细胞外Aβ蛋白的毒性。有研究表明,降低Tau蛋白的表达水平也可以作为治疗阿尔兹海默症的一个有效策略。如同其他没有酶功能的疾病相关蛋白类似,Tau蛋白的调控也是小分子化合物面临的一大难题,传统的小分子对于调控Tau蛋白来说作用十分有限。与直接靶向蛋白的小分子不同,PROTACs通过调动细胞中其他功能发挥作用,已经成为了靶向细胞内非酶蛋白并降低其表达水平的手段之一,并且可以作为选择性清除Tau蛋白聚集、治疗AD的新对策。
Lu等报道过一个基于肽的PROTAC分子Peptide-1,可以劫持Keap1-Cul3 E3泛素连接酶造成细胞内tau蛋白聚合物被泛素化标记并通过蛋白酶体降解。通过流式细胞仪和Western blotting实验分析,Peptide-1可以以时间依赖和浓度依赖的方式下调tau蛋白水平。
基于肽片段的PROTAC分子充分证明了针对Tau蛋白的降解策略确实可以有效降低Tau蛋白水平,避免毒性聚合物的形成。然而,肽的天然特性限制了上述Tau蛋白降解剂的临床使用。为了克服这一困难,Gray和Haggarty(Silva,M.C.,Ferguson,F.M.,Cai,Q.,etal.Targeted degradation of aberrant tau in frontotemporal dementia patient-derived neuronal cell models[J].Elife,2019,8:e45457.)实验室报道了第一个完全基于小分子的Tau蛋白PROTAC降解剂以期望用于治疗额颞叶痴呆(FTD)。作者利用已经用于临床检测的正电子发射断层成像(PET)探针18F-T807进行适当改造作为PROTAC分子的靶头部分、设计了通过泊马度胺作为E3连接酶配体招募E3连接酶,能够靶向泛素化标记Tau并且通过蛋白酶体将其降解的化合物QC-01-175。QC-01-175可以在FTD神经细胞模型中靶向降解多种形式的Tau蛋白,并且在小鼠动物模型中也能够区分疾病相关Tau蛋白与健康蛋白。全细胞蛋白组学分析表明QC-01-175具有比靶头更好的Tau蛋白选择性,包括其无法降解单胺氧化酶A、B。
目前,对于Tau蛋白降解剂的设计重点主要还是在于靶向Tau蛋白的靶头分子的设计。小分子Tau蛋白降解剂QC-01-175以PET探针18F-T807骨架作为Tau蛋白的靶向结合配体,但是只保留了化合物的母核结构,舍弃了发挥成像作用必需的放射性18F元素,由此只保留了PET探针对于Tau蛋白的靶向结合能力,但是对于成像功能却没有保留。
为此我们设想,如能在保留探针的靶向结合能力的同时,对于其成像能力也有所保留,在实现Tau蛋白被降解的同时,能够直观的观察到降解的过程,实现Tau的可视化降解必将为治疗阿尔兹海默症提供新的思路。
发明内容
本发明的目的在于提供一类靶向Tau蛋白的可视化降解化合物及其制备方法和应用,创造性的将荧光探针结构与PROTAC结构结合,该类化合物有明显的缠结Tau蛋白泛素化降解作用,可为阿尔兹海默症的治疗提供新的思路。
本发明的具体技术方案为:一类Tau蛋白可视化PROTAC降解化合物,该类化合物是将E3连接酶配体先与linker缩合制备出中间体,再与靶头探针分子相连后得到的,该类化合物的结构通式如下:
其中,E3连接酶配体为VHL配体、来那度胺配体或泊马度胺配体,E3连接酶配体R的结构式如下:
Linker为烷烃链或PEG链,结构式如下:
作为优选,选用的E3连接酶配体为VHL配体时所制备得到的PROTAC降解化合物D-12的结构式如下:
作为优选,选用的E3连接酶配体为泊马度胺配体时所制备得到的PROTAC降解化合物D-16的结构式如下:
进一步地,D-12的具体制备路线如下:
D-12的具体合成步骤为:
取AD-4-5溶于四氢呋喃中,搅拌下向其中加入DIPEA、DMAP,将反应液降温至0℃搅拌8-15min后加入HATU,自然升温至室温,搅拌0.5-2h后分批加入AD-L-V-2-2,室温下搅拌10-12h,监测到反应完全结束后,向反应液中加入水,用乙酸乙酯萃取,干燥后旋干,粗品经硅胶柱层析纯化,得到固体D-12。
进一步地,D-16的具体制备路线如下:
D-16的具体合成步骤为:
取AD-4-5溶于四氢呋喃中,搅拌下向其中加入DIPEA、DMAP,将反应液降温至0℃搅拌8-15min后加入HATU,自然升温至室温,搅拌0.5-2h后分批加入AD-L-B-2-2,室温下搅拌10-12h,监测到反应完全结束后,向反应液中加入水,用乙酸乙酯萃取,干燥后旋干,粗品经硅胶柱层析纯化,得到固体D-16。
进一步地,AD-L-V-2-2的合成路线如下:
a.DIPEA,HATU,DMAP,THF,25℃,2-3h;b.HCl/EtOH,CH3OH,25℃,6-10h.
AD-L-V-2-2的具体合成步骤为:
取AD-L-6-2溶解于四氢呋喃溶液中,室温搅拌下加入DIPEA、DMAP,将反应液置于冰-水浴中降温至0℃,分批加入HATU,继续搅拌8-15min后升至室温,室温下搅拌30-40min后,分3次加入VHL配体,室温下搅拌2-3h,反应完全后,将反应液减压浓缩,加水并用二氯甲烷萃取,干燥过滤后旋干,柱层析分离纯化,得到AD-L-V-2-1;
AD-L-V-2-1溶于甲醇中,加入HCl/EtOH溶液,25℃下搅拌6-10h,待反应完全后旋干反应液,得到AD-L-V-2-2。
进一步地,AD-L-B-2-2的合成路线如下:
a.Pomalidomide,DIPEA,DMF,90℃,10-12h;b.HCl/EtOH,CH3OH,25℃,6-10h.
AD-L-B-2-2的具体合成过程如下:
取2-(2,6-二氧代-哌啶-3-基)-4-氟基-异吲哚-1,3-二酮溶解于DMF中,搅拌下滴加DIPEA,继续加入N-Boc-戊二胺,缓慢升温至90℃搅拌10-12h,待反应完全后,将反应液缓慢冷却至室温,向反应液中加水,搅拌10-15min后用乙酸乙酯萃取3-4次,干燥过滤后旋干,柱层析分离纯化,得到AD-L-B-2-1;
AD-L-B-2-1溶于甲醇中,加入HCl/EtOH溶液,25℃下搅拌6-10h,待反应完全后旋干反应液,得到AD-L-B-2-2。
进一步地,AD-4-5的制备路线如下:
a.Dimethyl sulfate,K2CO3,DMF,-4℃,4-6h;b.POCl3,DMF,0℃;
c.((1,3-Dioxolan-2-yl)methyl)triphenylphosphonium bromide,CH3ONa/CH3OH(25%),THF;
d.Cyanoacetic Acid,K2CO3,MeOH,65℃;
AD-4-5的具体制备过程如下:
(1)向DMF中加入3,5-二甲氧基苯胺,搅拌,向反应液中分批加入碳酸钾,并置于冰-乙醇浴下将反应液降温至-4℃,用注射器向反应液中滴加硫酸二甲酯,并在0℃下搅拌2-3h后自然升至室温,继续2-3h后,向反应液中加水室温搅拌10-15min,之后用乙酸乙酯萃取、干燥、过滤后旋干得粗品、通过硅胶柱层析纯化,得到AD-4-1;
(2)取AD-4-1溶于DMF中,冰-水浴下搅拌使反应液降温至0℃,在0℃下滴加三氯氧磷,继续在0℃下搅拌10-15min后室温搅12-14h,向反应液中加水,搅拌30-60min后用乙酸乙酯萃取3次,干燥过滤后旋干,柱层析分离,得到AD-4-2;
(3)取(1,3-二氧环戊基-2-甲基)三苯基溴化膦,加入四氢呋喃溶液使其完全溶解,搅拌下加入质量浓度25%的甲醇钠的甲醇溶液,室温下搅拌15-20min后分批加入AD-4-2,缓慢升温至78℃搅拌12-16h,监测到原料没有剩余后,停止加热,向冷却至室温的反应液中加入水,搅拌10-15min,再通过4M的盐酸调节pH至中性,用乙酸乙酯萃取,干燥过滤后旋干,柱层析分离,得到AD-4-3;
(4)重复步骤(3)中的合成步骤,仅以AD-4-3替换步骤(3)中的AD-4-2进行反应,得到AD-4-4;
(5)取AD-4-4置于甲醇中,搅拌溶解,搅拌下分批加入碳酸钾,室温搅拌10-15min后分批加入腈乙酸,将反应液升温至65-70℃回流12-16h;反应完全后,缓慢冷却到室温并把甲醇旋干,加入乙酸乙酯溶解,氢氧化钾溶液调节pH至8,水洗,保留水相,向水相中加入1M盐酸调节pH至3,调节过程中有黑色的絮状固体析出,过滤、水洗并干燥得到AD-4-5。
上述公开的一类Tau蛋白可视化PROTAC降解化合物可应用在制备用于治疗阿尔兹海默症或由于Tau蛋白累积造成的相关疾病的Tau蛋白降解剂中,该类化合物与Tau蛋白结合时,除了发挥靶向降解Tau蛋白的功能,还能发出可检测的近红外荧光,从而使Tau蛋白的降解过程可视化。
相比于现有技术,本发明具有如下优点:
1.本申请在PROTAC降解剂分子设计环节的靶头选取上不仅保留靶头小分子与Tau蛋白的选择性结合作用,还赋予了其产生近红外荧光的全新功能,以使得目标化合物在与Tau蛋白结合时,除了发挥靶向降解Tau蛋白的功能,还能发出可检测的近红外荧光,从而使Tau蛋白的降解过程可视化;在随时间变化的荧光实验中得出,化合物D-16具有对p-Tau(S404)较强的降解能力,并且其自身的荧光强度变化与p-Tau水平变化规律基本相同,D-12与D-16均表现出了与p-Tau减少相似的荧光趋势,基本实现了可视化降解剂的设计,在近红外可视化条件下,期望对蛋白降解进程进行更为直观的观察与调控,甚至为疾病检测手段与治疗手段相结合提供了可能;
2.本申请提出的将蛋白降解和近红外荧光探针相结合的创新设计理念,为阿尔兹海默症或由于Tau蛋白累积造成的相关疾病的治疗提供新的思路;
3.通过荧光进行的化合物降解能力评价显示,D-12和D-16分别对p-Tau(S396)和p-Tau(S404)具有强烈的降解作用,在10μM给药24小时后,超过50%的p-Tau被降解,表现出了较强的对于磷酸化Tau蛋白的降解能力;
4.在荧光显微镜观察下,D-12与D-16对于磷酸化Tau蛋白的共定位作用表现较为强烈,D-12与D-16对于病理细胞内磷酸化Tau蛋白的靶向能力较强。
附图说明
图1为实施例一中制备的化合物D-11的H-NMR图;
图2为实施例一中制备的化合物D-11的C-NMR图;
图3为实施例二中制备的化合物D-12的H-NMR图;
图4为实施例二中制备的化合物D-12的C-NMR图;
图5为不同浓度下化合物D-11对N2a/APP细胞中p-Tau(S396)的降解活性图;其中,A.化合物D-11在不同浓度下对p-Tau(200kDa)和p-Tau(50kDa)的降解结果;B.化合物D-11对N2a/APP细胞中p-Tau(S396,50kDa)浓度影响柱状图;C.化合物D-11对N2a/APP细胞中p-Tau(S396,200kDa)浓度影响柱状图;注:分析方法采用One-way ANOVA,与不给药组相比,nsp>0.05;
图6为不同浓度下化合物D-12对N2a/APP细胞中p-Tau(S396)的降解活性图,其中A.化合物D-12在不同浓度下对p-Tau(200kDa)和p-Tau(50kDa)的降解结果;B.化合物D-12对N2a/APP细胞中p-Tau(S396,50kDa)浓度影响柱状图;C.化合物D-12对N2a/APP细胞中p-Tau(S396,200kDa)浓度影响柱状图;注:分析方法采用One-way ANOVA,与不给药组相比,*p<0.05,**p<0.01,nsp>0.05;
图7为实施例三中制备的化合物D-16的H-NMR图;
图8为实施例三中制备的化合物D-16的C-NMR图;
图9为实施例四中制备的化合物D-18的H-NMR图;
图10为实施例四中制备的化合物D-18的C-NMR图;
图11为不同浓度下化合物D-16对N2a/APP细胞中p-Tau(S396)的降解活性图;其中,A.化合物D-16在不同浓度下对p-Tau(200kDa)和p-Tau(50kDa)的降解结果;B.化合物D-16对N2a/APP细胞中p-Tau(S396,50kDa)浓度影响柱状图;C.化合物D-16对N2a/APP细胞中p-Tau(S396,200kDa)浓度影响柱状图。注:分析方法采用One-way ANOVA,与不给药组相比,*p<0.05,***p<0.0005,****p<0.0001;
图12为不同浓度下化合物D-18对N2a/APP细胞中p-Tau(S396)的降解活性图,其中,A.化合物D-18在不同浓度下对p-Tau(200kDa)和p-Tau(50kDa)的降解结果。B.化合物D-18对N2a/APP细胞中p-Tau(S396,50kDa)浓度影响柱状图。C.化合物D-18对N2a/APP细胞中p-Tau(S396,200kDa)浓度影响柱状图。注:分析方法采用One-way ANOVA,与不给药组相比,**p<0.01,*p<0.05,nsp>0.05;
图13是化合物D-12与Tau蛋白共定位图;化合物(1μM)作用24h后通过免疫荧光染色对三种类型的Tau蛋白进行绿色荧光标记:p-Tau(S396)、p-Tau(S404)和Tau-5,化合物呈红色荧光,放大倍数为40倍;
图14是化合物D-16与Tau蛋白共定位图,化合物(1μM)作用24h后通过免疫荧光染色对三种类型的Tau蛋白进行绿色荧光标记:p-Tau(S396)、p-Tau(S404)和Tau-5,化合物呈红色;
图15是化合物D-12、D-16给药后不同时间点下Tau蛋白水平荧光图;化合物给药浓度为10μM,图中绿色荧光为p-Tau(S404),蓝色荧光为细胞核,红色荧光为化合物,放大倍数为40倍;
图16是化合物D-12、D-16给药后不同时间点下Tau蛋白水平与化合物自身荧光水平统计学分析图;注:分析方法采用One-way ANOVA,***p<0.005,**p<0.01,*p<0.05,nsp>0.05。
具体实施方式
下面结合附图对本发明的技术方案作进一步的说明,但并不局限于此,凡是对本发明技术方案进行修改或者等同替换,而不脱离本发明技术方案的精神和范围,均应涵盖在本发明的保护范围中。
1.靶头部分的合成
以甲氧基取代的二甲胺基苯环为母核的靶头部分合成路线如下:
(a)Dimethyl sulfate,K2CO3,DMF,-4℃,4h;(b)POCl3,DMF,0℃,12h;(c)((1,3-Dioxolan-2-yl)methyl)triphenylphosphonium bromide,CH3ONa/CH3OH(25%),THF;(d)Cyanoacetic Acid,K2CO3,MeOH,70℃,12h;
具体制备过程如下:
向40mL DMF中加入3,5-二甲氧基苯胺(2.30g,15.00mmol),搅拌使其完全溶解。向反应液中分批加入碳酸钾(7.30g,60.00mmol),并置于冰-乙醇浴下将反应液降温至-4℃。然后用注射器向反应液中滴加硫酸二甲酯(4.2g,30.00mmol),并在0℃下搅拌2h后自然升至室温。继续2h后,向反应液中加水70mL室温搅拌10min,之后用乙酸乙酯(50mL)萃取,干燥、过滤后旋干得粗品。通过硅胶柱层析纯化,流动相为石油醚/乙酸乙酯=10/1,最终得到1.20g白色固体AD-4-1,收率为44%。
1H NMR(400MHz,Chloroform-d)δ5.91(s,3H),3.78(s,6H),2.93(s,6H).
取AD-4-1(360.00mg,2.00mmol)溶于20mL DMF中,冰-水浴下搅拌使反应液降温至0℃,在0℃下滴加三氯氧磷(640.00mg,4.2mmol),继续在0℃下搅拌10min后室温搅12h。向反应液中加入36mL水,搅拌30min后用乙酸乙酯(36mL)萃取3次,干燥过滤后旋干。柱层析分离,流动相为石油醚/乙酸乙酯=1/1,得到1.2g淡黄色固体AD-4-2,收率为46%。
1H NMR(400MHz,Chloroform-d)δ10.25(s,1H),5.73(s,2H),3.88(s,6H),3.09(s,6H).
取(1,3-二氧环戊基-2-甲基)三苯基溴化膦(2.58g,6.01mmol),加入15mL四氢呋喃溶液使其完全溶解,搅拌下加入质量浓度为25%的甲醇钠的甲醇溶液1.68mL,室温下搅拌15min后分批加入AD-4-2(630.00mg,3.01mmol),缓慢升温至78℃搅拌12h。反应过程中监测到原料没有剩余后,停止加热。向冷却至室温的反应液中加入水30mL,搅拌10min,再通过4M的盐酸调节pH至中性,用乙酸乙酯20mL萃取,干燥过滤后旋干。柱层析分离,流动相为石油醚/乙酸乙酯=2/1,得到黄色固体AD-4-3 460.00mg,产率为66%。再经过一次上述同样的合成步骤,得到AD-4-4,收率为60%。
1H NMR(400MHz,Chloroform-d)δ9.50(d,J=8.3Hz,1H),7.39(t,J=10.6Hz,1H),7.27(d,J=14.3Hz,2H),6.12(p,J=8.7Hz,1H),5.81(s,2H),3.88(s,6H),3.04(s,6H).
取AD-4-4(210.00mg,0.81mmol),置于10mL甲醇中,室温搅拌完全溶解,搅拌下分批加入碳酸钾(223.00mg,1.61mmol),室温搅拌10min后分批加入腈乙酸(205.00mg,2.41mmol)后将反应液升温至70℃回流12h。反应完全后,缓慢冷却到室温并把甲醇旋干。加入10mL乙酸乙酯溶解,用适当氢氧化钾溶液调节pH至8,之后用少量的水洗,保留水相。向水相中加入1M盐酸调节pH至3,调节过程中有黑色的絮状固体析出。过滤、水洗并干燥得到192.00mg AD-4-5粗品,呈黑色固体。
2.linker与E3连接酶配体的连接
主要有两种情况:(1)VHL配体和来那度胺的连接方法为配体的氨基与linker的羧基缩合;(2)泊马度胺作为配体时为泊马度胺的F取代与linker的氨基缩合。
VHL配体与linker的缩合路线
(a)DIPEA,HATU,DMAP,THF,25℃,2h;(c)HCl/EtOH,CH3OH,25℃,8h.
具体制备过程如下:
取AD-L-1-2(52.00mg,0.26mmol)溶解于8mL四氢呋喃溶液中,室温搅拌下加入N,N-二异丙基乙胺(DIPEA,117.00mg,0.90mmol)、4-二甲氨基吡啶(DMAP,38.00mg,0.31mmol),将反应液置于冰-水浴中降温至0℃,分批加入2-(7-氮杂苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯(HATU,196.00mg,0.51mmol),继续搅拌10min后升至室温,室温下搅拌30min后,分3次加入VHL配体(120.00mg,0.26mmol),室温下搅拌2h。待反应完全后,将反应液减压浓缩,加水20mL并用二氯甲烷20mL萃取,干燥过滤后旋干。柱层析分离纯化,得到白色固体AD-L-V-3-1 116.20mg,收率为74%。
1H NMR(400MHz,Chloroform-d)δ8.68(s,1H),7.58–7.47(m,1H),7.39–7.31(m,4H),7.12(t,J=8.2Hz,1H),4.98–4.83(m,1H),4.73(t,J=7.1Hz,1H),4.62–4.41(m,3H),4.33(dd,J=15.0,5.1Hz,1H),4.17–3.92(m,2H),3.61(d,J=10.0Hz,1H),3.19–2.93(m,2H),2.51(s,3H),2.28–2.09(m,4H),1.81–1.68(m,2H),1.42(s,9H),0.96(s,9H).
13C NMR(101MHz,Chloroform-d)δ173.61,171.88,171.09,156.61,150.32,148.40,138.26,131.63,130.81,129.44(2C),128.06(2C),79.53,70.12,58.58,58.22,56.87,43.13,39.46,36.28,34.85,33.08,28.40(3C),26.62,26.45(3C),16.04.
AD-L-V-3-1(150.00mg,0.24mmol)溶于10mL甲醇中,加入HCl/EtOH溶液0.5mL,25℃下搅拌8h,待反应完全后旋干反应液,得到白色固体AD-L-V-3-2粗品,未经纯化直接用于下一步反应。
AD-L-V-2-2的合成过程与AD-L-V-3-2相似,
取AD-L-6-2(52.00mg,0.26mmol)溶解于8mL四氢呋喃溶液中,室温搅拌下加入DIPEA(117.00mg,0.90mmol)、DMAP(38.00mg,0.31mmol),将反应液置于冰-水浴中降温至0℃,分批加入HATU(196.00mg,0.51mmol),继续搅拌10min后升至室温,室温下搅拌30min后,分3次加入VHL配体(120.00mg,0.26mmol),室温下搅拌2h。待反应完全后,将反应液减压浓缩,加水20mL并用二氯甲烷20mL萃取,干燥过滤后旋干。柱层析分离纯化,得到白色固体AD-L-V-2-1收率为76%。
1H NMR(400MHz,Chloroform-d)δ8.66(s,1H),7.47(t,1H),7.38–7.30(m,4H),6.43–6.29(m,1H),4.75–4.63(m,2H),4.56–4.47(m,3H),4.39–4.29(m,1H),4.20(t,1H),4.03(d,J=11.2Hz,1H),3.62(d,J=9.5Hz,1H),3.11–2.99(m,2H),2.60–2.55(m,2H),2.49(s,3H),2.44–2.35(m,1H),2.21–2.08(m,3H),1.47–1.36(m,11H),1.29–1.20(m,6H),0.93(s,9H).
13C NMR(101MHz,Chloroform-d)δ173.44,171.55,171.23,156.05,150.41,148.29,138.28,131.63,130.73,129.38(2C),127.95(2C),78.92,69.85,58.89,57.32,56.80,55.47,53.47,43.05,40.51,36.49,36.27,35.41,29.84,28.92,28.42(3C),26.42(3C),25.48,16.00.
AD-L-V-2-1(150.00mg,0.24mmol)溶于10mL甲醇中,加入HCl/EtOH溶液0.5mL,25℃下搅拌8h,待反应完全后旋干反应液,得到白色固体AD-L-V-2-2粗品,未经纯化直接用于下一步反应。
泊马度胺与linker缩合路线
(a)Pomalidomide,DIPEA,DMF,90℃,12h;(b)HCl/EtOH,CH3OH,25℃,8h.
具体合成过程如下:
取2-(2,6-二氧代-哌啶-3-基)-4-氟基-异吲哚-1,3-二酮(1.20g,4.35mmol)溶解于30mL DMF中,搅拌下滴加DIPEA(1.12g,8.69mmol),继续加入N-Boc-戊二胺(833.00mg,4.78mmol),缓慢升温至90℃搅拌12小时。待反应完全后,将反应液缓慢冷却至室温,向反应液中加水50mL,搅拌10min后用乙酸乙酯40mL萃取3次,干燥过滤后旋干。柱层析分离纯化,流动相为二氯甲烷/甲醇=60/1,得到黄色油状物AD-L-B-2-1,1.19g,收率为64%
1H NMR(400MHz,Chloroform-d)δ8.34(s,1H),7.49(t,1H),7.08(d,J=7.0Hz,1H),6.87(d,J=8.5Hz,1H),6.23(t,J=5.4Hz,1H),4.92(dd,J=11.9,5.3Hz,1H),4.59(t,1H),3.32–3.22(m,2H),3.18–3.07(m,2H),2.93–2.68(m,3H),2.17–2.09(m,1H),1.71–1.65(m,2H),1.58–1.49(m,2H),1.44(s,9H),1.30–1.23(m,1H).
13C NMR(101MHz,Chloroform-d)δ171.15,169.52,168.44,167.63,156.01,146.94,136.14,132.51,116.63,111.45,109.93,79.18,48.89,42.55,40.38,31.42,29.87,28.93,28.43(3C),24.16,22.81.
AD-L-B-2-1(150.00mg,0.35mmol)溶于10mL甲醇中,加入HCl/EtOH溶液0.5mL,25℃下搅拌8h,待反应完全后旋干反应液,得到黄绿色AD-L-B-2-2粗品,未经纯化直接用于下一步反应。
AD-L-B-3-2的合成方法与上述类似,具体中间体结构表征如下:
1H NMR(400MHz,Chloroform-d)δ8.97(s,1H),7.48(t,J=7.8Hz,1H),7.09(d,J=7.1Hz,1H),6.92(d,J=8.5Hz,1H),6.50(t,J=5.5Hz,1H),5.11(t,1H),5.01–4.90(m,1H),3.72–3.66(m,2H),3.58–3.53(m,2H),3.49–3.42(m,2H),3.36–3.30(m,2H),2.86–2.81(m,1H),2.80–2.75(m,1H),2.18–2.03(m,1H),1.42(s,9H).
1H NMR(400MHz,Methanol-d4)δ7.53(t,J=7.8Hz,1H),7.11–7.00(m,2H),5.04(dd,J=12.5,5.4Hz,1H),3.70(t,J=5.2Hz,2H),3.58–3.47(m,4H),2.90–2.78(m,3H),2.77–2.64(m,2H),2.14–2.06(m,1H),1.31–1.26(m,2H).
13C NMR(101MHz,Methanol-d4)δ173.30,170.23,169.43,167.86,146.85,135.82,132.45,116.93,110.73,110.00,71.60,69.05,41.77,40.66,30.83,29.35,22.40.
实施例一、靶头羧酸中间体与linker/E3配体复合物中间体缩合获得PROTAC终产分子
以VHL配体为E3配体的PROTACs合成路线
(2R,4R)-1-((S)-2-(4-((2E,4E,6E)-2-cyano-7-(4-(二甲氨基)-2,6-二甲氧基苯基))庚-2,4,6-三烯酰胺基)丁酰胺基)-3,3-二甲基丁酰基)-4-羟基-N-(4-(4-甲基噻唑-5-基)苄基)吡咯烷-2-甲酰胺D-11
D-11的具体合成步骤:
取AD-4-5(66.00mg,0.20mmol)溶于7mL四氢呋喃中,在搅拌下向其中加入DIPEA(105.00mg,0.81mmol)、DMAP(30.00mg,0.24mmol),将反应液降温至0℃搅拌10min后加入HATU(155.00mg,0.41mmol),自然升温至室温,搅拌半小时后分批加入AD-L-V-3-2(103.00mg,0.20mmol),室温下搅拌12h。监测到反应完全结束后,向反应液中加入水14mL,用乙酸乙酯14mL萃取,干燥后旋干。粗品经硅胶柱层析纯化,流动相为二氯甲烷/甲醇=70/1,得到固体D-1160.00mg,收率为36%。
1H NMR(400MHz,Chloroform-d)δ8.67(s,1H),7.92(d,J=12.3Hz,1H),7.42–7.30(m,7H),7.17–7.02(m,2H),6.74(dd,J=8.1Hz,1H),6.62(t,J=13.2Hz,1H),5.82(s,2H),4.81–4.73(m,1H),4.63–4.47(m,3H),4.33(dd,J=14.9,4.8Hz,1H),4.21(d,J=11.2Hz,1H),3.89(s,6H),3.64–3.57(m,2H),3.51–3.43(m,1H),3.40–3.33(m,1H),3.06(s,6H),2.52(s,3H),2.37–2.28(m,2H),2.16(s,1H),1.87(s,2H),1.39–1.32(m,1H),0.96(s,9H).
13C NMR(101MHz,Chloroform-d)δ173.70,172.07,170.75,162.32,161.02(2C),154.52,153.88,152.73,150.30,148.47,138.14,135.75,131.66,130.94,129.55(2C),128.18(2C),126.11,122.84,117.19,103.86,99.44,88.08(2C),70.22,58.42,58.22,56.83,55.50(2C),43.27,40.31(2C),39.83,35.66,34.52,33.59,26.48(3C),25.00,16.09.
HRMS(ESI)m/z calcd for C44H56N7O7S(M+H)+826.3959;found,826.3962.
Purity:98.87%.
实施例二、靶头羧酸中间体与linker/E3配体复合物中间体缩合获得PROTAC终产分子
以VHL配体为E3配体的PROTACs合成路线
(2R,4R)-1-((S)-2-(8-((2E,4E,6E)-2-氰基-7-(4-(二甲氨基)-2,6-二甲氧基苯基))庚-2,4,6-三烯酰胺基)辛酰胺基)-3,3-二甲基丁酰基)-4-羟基-N-)4-(4-甲基噻唑-5-基)苄基)吡咯烷-2-甲酰胺D-12;
D-12的具体合成步骤:
取AD-4-5(66.00mg,0.20mmol)溶于7mL四氢呋喃中,在搅拌下向其中加入DIPEA(105.00mg,0.81mmol)、DMAP(30.00mg,0.24mmol),将反应液降温至0℃搅拌10分钟后加入HATU(155.00mg,0.41mmol),自然升温至室温,搅拌半小时后分批加入AD-L-V-2-2(114.00mg,0.20mmol),室温下搅拌12h。监测到反应完全结束后,向反应液中加入水14mL,用乙酸乙酯14mL萃取,干燥后旋干。粗品经硅胶柱层析纯化,流动相为二氯甲烷/甲醇=70/1,得到固体D-12,77.00mg,收率为43%。
结构确证数据如下:
1H NMR(400MHz,Chloroform-d)δ8.68(s,1H),7.91(d,J=12.3Hz,1H),7.46–7.29(m,7H),7.04(dd,J=13.9,9.6Hz,1H),6.62(t,J=13.2Hz,1H),6.21–6.11(m,2H),5.83(s,2H),4.75(t,J=8.0Hz,1H),4.63–4.50(m,3H),4.35(dd,J=15.0,5.2Hz,1H),4.15(d,J=11.4Hz,1H),3.89(s,6H),3.59(d,J=8.6Hz,1H),3.39–3.31(m,2H),3.06(s,6H),2.52(s,3H),2.23–2.12(m,3H),1.63–1.51(m,4H),1.36–1.27(m,8H),0.94(s,9H).
13C NMR(101MHz,Chloroform-d)δ173.88,172.02,170.68,161.70,160.97(2C),154.09,153.63,152.66,150.29,148.40,138.11,135.52,131.57,130.94,129.54(2C),128.12(2C),126.16,122.93,117.35,99.67,88.11(2C),70.09,58.31,57.49,56.69,55.48(2C),43.25,40.32(2C),40.18,36.23,35.77,34.70,29.70,29.30,28.67,26.54,26.42(3C),25.28,16.05.
HRMS(ESI)m/z calcd for C48H64N7O7S(M+H)+882.4526;found,882.4588.
图5和图6分别反映了不同浓度下化合物D-11和D-12对N2a/APP细胞中p-Tau(S396)的降解活性;从图中可以看出,与D-11相比,D-12增长linker至八个原子长度同时保留VHL作为E3连接酶配体,对磷酸化Tau降解能力显著提升,特别是对于单体p-Tau。在给药浓度为10μM时即可使p-Tau(50kd)降解至正常细胞水平,并且随着给药浓度的增加,D-12对于p-Tau降解活性逐渐增强,在给药10μM时可将病理细胞中单体p-Tau水平降低至正常细胞的一半。
实施例三、靶头羧酸中间体与linker/E3配体复合物中间体缩合获得PROTAC终产分子
以泊马度胺为E3配体的PROTACs合成路线
(2E,4E,6E)-2-氰基-7-(4-(二甲氨基)-2,6-二甲氧基苯基)-N-(5-((2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-4-基)氨基)戊基)庚-2,4,6-三烯酰胺D-16;
D-16的具体合成步骤:
取AD-4-5(66.00mg,0.20mmol)溶于7mL四氢呋喃中,在搅拌下向其中加入DIPEA(105.00mg,0.81mmol)、DMAP(30.00mg,0.24mmol),将反应液降温至0℃搅拌10分钟后加入HATU(155.00mg,0.41mmol),自然升温至室温,搅拌半小时后分批加入AD-L-B-2-2(71.00mg,0.20mmol),室温下搅拌12h。监测到反应完全结束后,向反应液中加入水14mL,用14mL乙酸乙酯萃取,干燥后旋干。粗品经硅胶柱层析纯化,流动相为二氯甲烷/甲醇=70/1,得到固体D-16,57.00mg,收率为42%。
结构确证数据如下:
1H NMR(400MHz,DMSO-d6)δ11.10(s,1H),8.09(t,J=5.3Hz,1H),7.81(d,J=11.8Hz,1H),7.58(t,J=7.6Hz,1H),7.30–7.16(m,3H),7.11(d,J=8.5Hz,1H),7.02(d,J=6.3Hz,1H),6.57–6.43(m,2H),5.93(s,2H),5.05(dd,J=12.9,5.0Hz,1H),3.86(s,6H),3.65–3.58(m,1H),3.20–3.11(m,3H),3.03(s,6H),2.90–2.81(m,2H),2.06–1.98(m,1H),1.63–1.56(m,2H),1.55–1.47(m,2H),1.38–1.31(m,2H).
13C NMR(101MHz,DMSO-d6)δ173.30,170.59,169.41,167.79,161.82,160.93(2C),153.18,153.06,152.38,147.58,146.88,136.76,135.18,132.66,125.78,122.56,117.70,110.85,109.47,102.97,101.95,88.69(2C),55.98(2C),54.05,48.99,42.31,31.62,30.28,29.50,28.87,22.62,18.55,17.19.
HRMS(ESI)m/z calcd for C36H41N6O7(M+H)+669.3093;found,669.3037.
Purity:97.20%.
实施例四、靶头羧酸中间体与linker/E3配体复合物中间体缩合获得PROTAC终产分子
以泊马度胺为E3配体的PROTACs合成路线
(2E,4E,6E)-2-氰基-7-(4-(二甲氨基)-2,6-二甲氧基苯基)-N-(2-(2-((2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-4-基)氨基)乙氧基)乙基)庚-2,4,6-三烯酰胺D-18;
D-18的具体合成步骤:
取AD-4-5(66.00mg,0.20mmol)溶于7mL四氢呋喃中,在搅拌下向其中加入DIPEA(105.00mg,0.81mmol)、DMAP(30.00mg,0.24mmol),将反应液降温至0℃搅拌10分钟后加入HATU(155.00mg,0.41mmol),自然升温至室温,搅拌半小时后分批加入AD-L-B-3-2(72.00mg,0.20mmol),室温下搅拌12h。监测到反应完全结束后,向反应液中加入水14mL,用乙酸乙酯14mL萃取,干燥后旋干。粗品经硅胶柱层析纯化,流动相为二氯甲烷/甲醇=70/1,得到固体D-1854.00mg,收率为40%。
结构确证数据如下:
1H NMR(400MHz,DMSO-d6)δ11.09(s,1H),8.10–8.04(m,1H),7.81(dd,J=12.0,2.1Hz,1H),7.58(t,J=8.0Hz,1H),7.43–7.34(m,1H),7.28–7.20(m,2H),7.19–7.12(m,2H),7.03(dd,J=7.0,2.0Hz,1H),6.66–6.59(m,1H),6.47(t,J=12.5Hz,1H),5.93(s,2H),5.04(dd,J=13.0,5.5Hz,1H),3.86(s,6H),3.63(t,J=5.8Hz,2H),3.56–3.50(m,2H),3.48(d,J=5.8Hz,2H),3.03(s,6H),2.93–2.82(m,2H),2.58(d,J=16.7Hz,1H),2.08–1.95(m,1H),0.85(t,J=5.9Hz,1H).
13C NMR(101MHz,DMSO-d6)δ173.27,170.56,169.4,167.77,162.08,160.97(2C),158.82,153.22,146.88,136.72,135.36,132.55,125.76,124.80,122.51,117.93,116.89,111.15,109.71,102.99,101.62,88.69(2C),69.13,55.99(2C),49.02,42.11,34.96,31.62,30.29(2C),18.55,17.20.
HRMS(ESI)m/z calcd for C35H39N6O8(M+H)+671.2808;found,671.2829.
Purity:99.99%.
图11和图12分别反映了不同浓度下化合物D-16和D-18对N2a/APP细胞中p-Tau(S396)的降解活性;从图中可以看出,D-16对单体和缠结磷酸化Tau均表现出了很强的降解活性,在给药10μM时即可将两种Tau都降解到与正常细胞相当的水平,实现了在低浓度给药条件下即对目标靶蛋白有较好的降解作用。并且从数据图中可以看出,D-16不会将p-Tau降解至明显低于正常细胞的水平,因而其可能会产生相比其他化合物更小的副作用。
D-18对p-Tau的降解能力稍弱,不管是对于单体Tau还是缠结Tau都需要在给药浓度10μM以上才有降解作用。
当化合物通过VHL配体招募VHL作为E3连接酶时(D-11、D-12),并没有达到预期的降解活性,特别是对于缠结p-Tau的降解能力较弱,要在给药浓度10μM条件下才可以将缠结的Tau降解至正常水平。但是D-12对于单体Tau表现出了不错的降解活性,10μM浓度即可将单体磷酸化Tau降解至正常水平。
通过WB实验分析化合物对于单体p-Tau和缠结p-Tau的降解能力,D-16对于单体p-Tau与缠结p-Tau均表现出了突出的降解能力,在10μM给药浓度下即可将两者降解至正常水平。
在分子水平,化合物的紫外最大吸收波长在480-500nm,荧光最大发射波长在650-700nm,具有较大的斯托克斯位移。
细胞水平荧光活性
图13和14分别是化合物D-12、D-16与Tau蛋白共定位图,从数据结果可以看出,D-12对p-Tau(S396)表现出了很强的共定位能力,体现了其对p-Tau(S396)的强靶向能力。而D-12对于p-Tau(S404)与Tau-5的靶向能力明显减弱。降解能力最强的化合物D-16也表现出了较强的Tau蛋白靶向性,特别是对于p-Tau(S396)和全长Tau,对于p-Tau(S404)表现出的共定位稍弱。综上所述,D-12与D-16对于磷酸化Tau蛋白的共定位作用表现较为强烈,可以观察到merge图中出现了明显的黄色,这也说明D-12与D-16对于病理细胞内磷酸化Tau蛋白的靶向能力较强。
可视化降解活性评价
从图15和图16可以看出,在不同时间点的细胞内荧光实验结果中,D-12和D-16都表现出了随时间变化的降解效果,给药2小时后,化合物进入细胞并开始发挥降解作用。D-12在给药12小时后,化合物在细胞内浓度最大(红色荧光最强),在12到24小时内,化合物明显表现出了p-Tau蛋白的降解能力(绿色荧光减弱,蓝色荧光增强)。D-16在8到12小时内对p-Tau降解作用最为强烈,并且在给药24小时后,只观察到微弱的红色荧光,说明化合物D-16在给药24小时后基本被代谢完全。
通过荧光进行的化合物降解能力评价显示,D-12和D-16分别对p-Tau(S396)和p-Tau(S404)具有强烈的降解作用,在10μM给药24小时后,超过50%的p-Tau被降解,表现出了较强的对于磷酸化Tau蛋白的降解能力。在随时间变化的荧光实验中得出,化合物D-16具有对p-Tau(S404)较强的降解能力,并且其自身的荧光强度变化与p-Tau水平变化规律基本相同,D-12与D-16均表现出了与p-Tau减少相似的荧光趋势,基本实现了可视化降解剂的设计。
以上所述仅为本发明的实施例,并非因此限制本发明的专利范围,凡是利用本发明说明书及附图内容所作的等效结构或等效流程变换,或直接或间接运用在其他相关的技术领域,均同理包括在本发明的专利保护范围内。
Claims (9)
6.如权利要求4所述的一类Tau蛋白可视化PROTAC降解化合物的制备方法,其特征在于,AD-L-V-2-2的合成路线如下:
a.DIPEA,HATU,DMAP,THF,25℃,2-3h;b.HCl/EtOH,CH3OH,25℃,6-10h.
AD-L-V-2-2的具体合成步骤为:
取AD-L-6-2溶解于四氢呋喃溶液中,室温搅拌下加入DIPEA、DMAP,将反应液置于冰-水浴中降温至0℃,分批加入HATU,继续搅拌8-15min后升至室温,室温下搅拌30-40min后,分3次加入VHL配体,室温下搅拌2-3h,反应完全后,将反应液减压浓缩,加水并用二氯甲烷萃取,干燥过滤后旋干,柱层析分离纯化,得到AD-L-V-2-1;
AD-L-V-2-1溶于甲醇中,加入HCl/EtOH溶液,25℃下搅拌6-10h,待反应完全后旋干反应液,得到AD-L-V-2-2。
7.如权利要求5所述的一类Tau蛋白可视化PROTAC降解化合物的制备方法,其特征在于,AD-L-B-2-2的合成路线如下:
a.Pomalidomide,DIPEA,DMF,90℃,10-12h;b.HCl/EtOH,CH3OH,25℃,6-10h.
AD-L-B-2-2的具体合成过程如下:
取2-(2,6-二氧代-哌啶-3-基)-4-氟基-异吲哚-1,3-二酮溶解于DMF中,搅拌下滴加DIPEA,继续加入N-Boc-戊二胺,缓慢升温至90℃搅拌10-12h,待反应完全后,将反应液缓慢冷却至室温,向反应液中加水,搅拌10-15min后用乙酸乙酯萃取3-4次,干燥过滤后旋干,柱层析分离纯化,得到AD-L-B-2-1;
AD-L-B-2-1溶于甲醇中,加入HCl/EtOH溶液,25℃下搅拌6-10h,待反应完全后旋干反应液,得到AD-L-B-2-2。
8.如权利要求4-5中任一项所述的一类Tau蛋白可视化PROTAC降解化合物的制备方法,其特征在于,AD-4-5的制备路线如下:
a.Dimethyl sulfate,K2CO3,DMF,-4℃,4-6h;b.POCl3,DMF,0℃;
c.((1,3-Dioxolan-2-yl)methyl)triphenylphosphonium bromide,CH3ONa/CH3OH(25%),THF;
d.Cyanoacetic Acid,K2CO3,MeOH,65℃;
AD-4-5的具体制备过程如下:
(1)向DMF中加入3,5-二甲氧基苯胺,搅拌,向反应液中分批加入碳酸钾,并置于冰-乙醇浴下将反应液降温至-4℃,用注射器向反应液中滴加硫酸二甲酯,并在0℃下搅拌2-3h后自然升至室温,继续2-3h后,向反应液中加水室温搅拌10-15min,之后用乙酸乙酯萃取、干燥、过滤后旋干得粗品、通过硅胶柱层析纯化,得到AD-4-1;
(2)取AD-4-1溶于DMF中,冰-水浴下搅拌使反应液降温至0℃,在0℃下滴加三氯氧磷,继续在0℃下搅拌10-15min后室温搅12-14h,向反应液中加水,搅拌30-60min后用乙酸乙酯萃取3次,干燥过滤后旋干,柱层析分离,得到AD-4-2;
(3)取(1,3-二氧环戊基-2-甲基)三苯基溴化膦,加入四氢呋喃溶液使其完全溶解,搅拌下加入质量浓度25%的甲醇钠的甲醇溶液,室温下搅拌15-20min后分批加入AD-4-2,缓慢升温至78℃搅拌12-16h,监测到原料没有剩余后,停止加热,向冷却至室温的反应液中加入水,搅拌10-15min,再通过4M的盐酸调节pH至中性,用乙酸乙酯萃取,干燥过滤后旋干,柱层析分离,得到AD-4-3;
(4)重复步骤(3)中的合成步骤,仅以AD-4-3替换步骤(3)中的AD-4-2进行反应,得到AD-4-4;
(5)取AD-4-4置于甲醇中,搅拌溶解,搅拌下分批加入碳酸钾,室温搅拌10-15min后分批加入腈乙酸,将反应液升温至65-70℃回流12-16h;反应完全后,缓慢冷却到室温并把甲醇旋干,加入乙酸乙酯溶解,氢氧化钾溶液调节pH至8,水洗,保留水相,向水相中加入1M盐酸调节pH至3,调节过程中有黑色的絮状固体析出,过滤、水洗并干燥得到AD-4-5。
9.如权利要求1-3中任一项所述的一类Tau蛋白可视化PROTAC降解化合物在制备用于治疗阿尔兹海默症或由于Tau蛋白累积造成的相关疾病的Tau蛋白降解剂方面的应用,其特征在于,该类化合物与Tau蛋白结合时,除了发挥靶向降解Tau蛋白的功能,还能发出可检测的近红外荧光,从而使Tau蛋白的降解过程可视化。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202210387786.1A CN114736264B (zh) | 2022-04-14 | 2022-04-14 | Tau蛋白可视化PROTAC降解化合物及其制备方法和应用 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202210387786.1A CN114736264B (zh) | 2022-04-14 | 2022-04-14 | Tau蛋白可视化PROTAC降解化合物及其制备方法和应用 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN114736264A true CN114736264A (zh) | 2022-07-12 |
CN114736264B CN114736264B (zh) | 2024-04-02 |
Family
ID=82281160
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202210387786.1A Active CN114736264B (zh) | 2022-04-14 | 2022-04-14 | Tau蛋白可视化PROTAC降解化合物及其制备方法和应用 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN114736264B (zh) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN115475164A (zh) * | 2022-08-22 | 2022-12-16 | 西安交通大学 | 一种可降解PDGFR-β的蛋白降解靶向嵌合体及其制备方法和应用 |
Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN110234646A (zh) * | 2016-11-01 | 2019-09-13 | 阿尔维纳斯股份有限公司 | 靶向PROTAC的Tau蛋白及相关使用方法 |
CN111518215A (zh) * | 2019-02-01 | 2020-08-11 | 华中科技大学 | 特异性降解tau蛋白的嵌合体及其编码基因以及它们的应用 |
WO2021133037A1 (ko) * | 2019-12-26 | 2021-07-01 | 연세대학교 산학협력단 | 피롤리딘 유도체 및 이를 포함하는 베타-아밀로이드 또는 타우 단백질 관련 질병의 예방 또는 치료용 약학 조성물 |
CN113307793A (zh) * | 2020-08-27 | 2021-08-27 | 杭州医学院 | 基于CRBN配体诱导Tau蛋白降解的化合物及其制备方法、药物组合物和应用 |
WO2021178355A1 (en) * | 2020-03-02 | 2021-09-10 | President And Fellows Of Harvard College | New inhibitors for the keap1-nrf2 protein-protein interaction |
-
2022
- 2022-04-14 CN CN202210387786.1A patent/CN114736264B/zh active Active
Patent Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN110234646A (zh) * | 2016-11-01 | 2019-09-13 | 阿尔维纳斯股份有限公司 | 靶向PROTAC的Tau蛋白及相关使用方法 |
CN111518215A (zh) * | 2019-02-01 | 2020-08-11 | 华中科技大学 | 特异性降解tau蛋白的嵌合体及其编码基因以及它们的应用 |
WO2021133037A1 (ko) * | 2019-12-26 | 2021-07-01 | 연세대학교 산학협력단 | 피롤리딘 유도체 및 이를 포함하는 베타-아밀로이드 또는 타우 단백질 관련 질병의 예방 또는 치료용 약학 조성물 |
WO2021178355A1 (en) * | 2020-03-02 | 2021-09-10 | President And Fellows Of Harvard College | New inhibitors for the keap1-nrf2 protein-protein interaction |
CN113307793A (zh) * | 2020-08-27 | 2021-08-27 | 杭州医学院 | 基于CRBN配体诱导Tau蛋白降解的化合物及其制备方法、药物组合物和应用 |
Non-Patent Citations (2)
Title |
---|
M CATARINA SILVA等: "Targeted degradation of aberrant tau in frontotemporal dementia patient-derived neuronal cell models", ELIFE, vol. 8, no. 45457, pages 1 - 31 * |
WEIJIN WANG等: "A novel small-molecule PROTAC selectively promotes tau clearance to improve cognitive functions in Alzheimer-like models", THERANOSTICS等, vol. 11, no. 11, pages 5279 - 5295, XP093137050, DOI: 10.7150/thno.55680 * |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN115475164A (zh) * | 2022-08-22 | 2022-12-16 | 西安交通大学 | 一种可降解PDGFR-β的蛋白降解靶向嵌合体及其制备方法和应用 |
Also Published As
Publication number | Publication date |
---|---|
CN114736264B (zh) | 2024-04-02 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US5582814A (en) | 1-(p-n-butylbenzyl) DTPA for magnetic resonance imaging | |
US20030129579A1 (en) | Multi-use multimodal imaging chelates | |
EP3074388A1 (en) | Dotam derivatives for therapeutic use | |
CN109963838A (zh) | 二聚造影剂 | |
CN114736264A (zh) | Tau蛋白可视化PROTAC降解化合物及其制备方法和应用 | |
JPH09503500A (ja) | ジキレート化剤としてのポリアザシクロアルカン | |
CN103911017A (zh) | 菁染料化合物及其制备方法、用于光动力学疗法的双重功能剂及其制备方法 | |
US20140294873A1 (en) | Small molecule screen for inhibitors of nfat: ap-1: dna interactions | |
US6239145B1 (en) | Nitroxyl compounds and drugs and reagents containing the same as the active ingredient | |
CN102167813B (zh) | 一种荧光示踪纳米磁共振成像造影剂 | |
CN114835710B (zh) | 双功能大环螯合物、偶联物、金属络合物及其应用 | |
CN114436960B (zh) | 一种用于靶向治疗的光活性核酸适体偶联药物及其制备方法与用途 | |
CN114437128A (zh) | 一种胆碱磷酸修饰的紫杉醇药物及其制备方法和应用 | |
WO2018067551A1 (en) | Texaphyrin and antitumor antibiotic conjugates | |
KR20190111376A (ko) | 신규한 구조를 갖는 화합물, 이를 포함하는 항염증제 및 기질 금속 단백질분해효소-9 억제제 | |
CN109602920B (zh) | 一类树枝状分子影像探针及其制备方法 | |
CN114632079B (zh) | 一种基于青蒿素的铁池靶向分子影像探针制备及其应用 | |
CN102190644A (zh) | 手性3-羟基吡啶-4-酮类衍生物及其合成和用途 | |
KR101389258B1 (ko) | 니트로이미다졸-아미노산 핵 저산소증 조영제 및 그 전구체 | |
CN100355806C (zh) | 端氨基嵌段聚氧乙烯改性的钆配合物及其合成方法 | |
WO2004069787A1 (de) | Enantiomerenreines (4s,8s)- und (4r, 8r)-4-p-nitrobenzyl-8-methyl-3, 6, 9-triaza-3n, 6n, 9n-tricarboxymethyl-1, 11-undecandisäure und deren abkömmlinge, verfahren zu deren herstellung und verwendung zur herstellung pharmazeutischer mittel | |
CN104436219B (zh) | 一种含硝基咪唑基的水溶性磁共振成像造影剂及其制备方法 | |
KR102659229B1 (ko) | 가돌리늄계 화합물, 이를 포함하는 mri 조영제 | |
CN111214667B (zh) | 一种具有核-壳结构的电响应胶束磁共振探针及其制备方法和应用 | |
CN100404585C (zh) | 一种聚氧乙烯类嵌段聚合物配体及其合成方法 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
GR01 | Patent grant | ||
GR01 | Patent grant |