CN117820416A - 一种Hsp90 PROTAC化合物及其制备方法与在制备抗宫颈癌药物中的应用 - Google Patents
一种Hsp90 PROTAC化合物及其制备方法与在制备抗宫颈癌药物中的应用 Download PDFInfo
- Publication number
- CN117820416A CN117820416A CN202311816125.7A CN202311816125A CN117820416A CN 117820416 A CN117820416 A CN 117820416A CN 202311816125 A CN202311816125 A CN 202311816125A CN 117820416 A CN117820416 A CN 117820416A
- Authority
- CN
- China
- Prior art keywords
- amino
- dimethyl
- phenyl
- compound
- indazol
- 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
- 150000001875 compounds Chemical class 0.000 title claims abstract description 67
- 206010008342 Cervix carcinoma Diseases 0.000 title claims abstract description 38
- 208000006105 Uterine Cervical Neoplasms Diseases 0.000 title claims abstract description 38
- 201000010881 cervical cancer Diseases 0.000 title claims abstract description 38
- 238000002360 preparation method Methods 0.000 title claims abstract description 35
- 239000003560 cancer drug Substances 0.000 title claims abstract description 9
- -1 lenalidomide carboxylic acid derivatives Chemical class 0.000 claims abstract description 59
- JGFZNNIVVJXRND-UHFFFAOYSA-N N,N-Diisopropylethylamine (DIPEA) Chemical compound CCN(C(C)C)C(C)C JGFZNNIVVJXRND-UHFFFAOYSA-N 0.000 claims abstract description 24
- NVGFSTMGRRADRG-IOJSEOPQSA-N chembl553939 Chemical compound CS(O)(=O)=O.O=C1CC(C)(C)CC2=C1C(C(F)(F)F)=NN2C(C=1)=CC=C(C(N)=O)C=1N[C@H]1CC[C@H](OC(=O)CN)CC1 NVGFSTMGRRADRG-IOJSEOPQSA-N 0.000 claims abstract description 16
- 239000003814 drug Substances 0.000 claims abstract description 14
- 239000007821 HATU Substances 0.000 claims abstract description 12
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 10
- 239000003513 alkali Substances 0.000 claims abstract 3
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Natural products NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 claims description 25
- 239000004471 Glycine Substances 0.000 claims description 24
- 102100034051 Heat shock protein HSP 90-alpha Human genes 0.000 claims description 17
- 101001016865 Homo sapiens Heat shock protein HSP 90-alpha Proteins 0.000 claims description 16
- 230000015556 catabolic process Effects 0.000 claims description 12
- 238000006731 degradation reaction Methods 0.000 claims description 12
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 claims description 9
- 125000004214 1-pyrrolidinyl group Chemical group [H]C1([H])N(*)C([H])([H])C([H])([H])C1([H])[H] 0.000 claims description 8
- 125000003917 carbamoyl group Chemical group [H]N([H])C(*)=O 0.000 claims description 8
- 125000000113 cyclohexyl group Chemical group [H]C1([H])C([H])([H])C([H])([H])C([H])(*)C([H])([H])C1([H])[H] 0.000 claims description 8
- 239000003446 ligand Substances 0.000 claims description 7
- GOTYRUGSSMKFNF-UHFFFAOYSA-N lenalidomide Chemical class C1C=2C(N)=CC=CC=2C(=O)N1C1CCC(=O)NC1=O GOTYRUGSSMKFNF-UHFFFAOYSA-N 0.000 claims description 6
- 238000000034 method Methods 0.000 claims description 6
- 150000003839 salts Chemical class 0.000 claims description 6
- 230000009471 action Effects 0.000 claims description 3
- 125000002801 octanoyl group Chemical group C(CCCCCCC)(=O)* 0.000 claims description 3
- 238000006482 condensation reaction Methods 0.000 claims description 2
- 125000003074 decanoyl group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C(*)=O 0.000 claims description 2
- 230000000593 degrading effect Effects 0.000 claims description 2
- 239000003937 drug carrier Substances 0.000 claims description 2
- 125000003104 hexanoyl group Chemical group O=C([*])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 claims description 2
- 239000000546 pharmaceutical excipient Substances 0.000 claims description 2
- 238000004519 manufacturing process Methods 0.000 claims 3
- 229940124657 anti-cervical cancer agent Drugs 0.000 claims 1
- 238000006243 chemical reaction Methods 0.000 abstract description 54
- 239000002994 raw material Substances 0.000 abstract description 23
- 230000000694 effects Effects 0.000 abstract description 22
- 229940079593 drug Drugs 0.000 abstract description 8
- 229960004942 lenalidomide Drugs 0.000 abstract description 6
- 150000001732 carboxylic acid derivatives Chemical class 0.000 abstract description 5
- 239000003481 heat shock protein 90 inhibitor Substances 0.000 abstract description 4
- 230000005764 inhibitory process Effects 0.000 abstract description 3
- 150000001408 amides Chemical class 0.000 abstract description 2
- 238000006555 catalytic reaction Methods 0.000 abstract 1
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 90
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 31
- 239000007858 starting material Substances 0.000 description 29
- 239000007787 solid Substances 0.000 description 17
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 15
- 230000008034 disappearance Effects 0.000 description 14
- 239000012265 solid product Substances 0.000 description 13
- 210000004027 cell Anatomy 0.000 description 12
- 102000004169 proteins and genes Human genes 0.000 description 12
- 108090000623 proteins and genes Proteins 0.000 description 12
- JQYVOBBMCVLZAG-UHFFFAOYSA-N 2-bromo-4-[6,6-dimethyl-4-oxo-3-(trifluoromethyl)-5,7-dihydroindazol-1-yl]benzonitrile Chemical compound O=C1CC(C)(C)CC2=C1C(C(F)(F)F)=NN2C1=CC=C(C#N)C(Br)=C1 JQYVOBBMCVLZAG-UHFFFAOYSA-N 0.000 description 11
- 235000019439 ethyl acetate Nutrition 0.000 description 11
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 10
- 239000012071 phase Substances 0.000 description 10
- 238000003756 stirring Methods 0.000 description 10
- 238000004440 column chromatography Methods 0.000 description 9
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 8
- 238000012544 monitoring process Methods 0.000 description 8
- 238000011282 treatment Methods 0.000 description 8
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 description 7
- 239000000047 product Substances 0.000 description 7
- 238000002390 rotary evaporation Methods 0.000 description 7
- 239000000243 solution Substances 0.000 description 7
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 6
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical class [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 6
- 238000010609 cell counting kit-8 assay Methods 0.000 description 6
- 238000011161 development Methods 0.000 description 6
- 238000005516 engineering process Methods 0.000 description 6
- 238000000338 in vitro Methods 0.000 description 6
- HEDRZPFGACZZDS-MICDWDOJSA-N Trichloro(2H)methane Chemical compound [2H]C(Cl)(Cl)Cl HEDRZPFGACZZDS-MICDWDOJSA-N 0.000 description 5
- 238000001035 drying Methods 0.000 description 5
- 239000003208 petroleum Substances 0.000 description 5
- 229940124823 proteolysis targeting chimeric molecule Drugs 0.000 description 5
- DJNCXSGGAMADNN-UHFFFAOYSA-N tert-butyl n-[(4-bromophenyl)methyl]carbamate Chemical compound CC(C)(C)OC(=O)NCC1=CC=C(Br)C=C1 DJNCXSGGAMADNN-UHFFFAOYSA-N 0.000 description 5
- 229940126585 therapeutic drug Drugs 0.000 description 5
- ZFVRYNYOPQZKDG-UHFFFAOYSA-N 4-[6,6-dimethyl-4-oxo-3-(trifluoromethyl)-5,7-dihydroindazol-1-yl]-2-[(4-hydroxycyclohexyl)amino]benzamide Chemical compound O=C1CC(C)(C)CC2=C1C(C(F)(F)F)=NN2C(C=1)=CC=C(C(N)=O)C=1NC1CCC(O)CC1 ZFVRYNYOPQZKDG-UHFFFAOYSA-N 0.000 description 4
- OFQAFLDKWBQAJI-UHFFFAOYSA-N 6,6-dimethyl-3-(trifluoromethyl)-5,7-dihydro-1h-indazol-4-one Chemical compound O=C1CC(C)(C)CC2=C1C(C(F)(F)F)=NN2 OFQAFLDKWBQAJI-UHFFFAOYSA-N 0.000 description 4
- XKRFYHLGVUSROY-UHFFFAOYSA-N Argon Chemical compound [Ar] XKRFYHLGVUSROY-UHFFFAOYSA-N 0.000 description 4
- QVZWBGUVSPTOQF-UHFFFAOYSA-N CC=1N=CSC=1C1=CC=C(CNC(OC(C)(C)C)=O)C=C1 Chemical compound CC=1N=CSC=1C1=CC=C(CNC(OC(C)(C)C)=O)C=C1 QVZWBGUVSPTOQF-UHFFFAOYSA-N 0.000 description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 4
- 229910021529 ammonia Inorganic materials 0.000 description 4
- 239000006285 cell suspension Substances 0.000 description 4
- DQLATGHUWYMOKM-UHFFFAOYSA-L cisplatin Chemical compound N[Pt](N)(Cl)Cl DQLATGHUWYMOKM-UHFFFAOYSA-L 0.000 description 4
- 238000000746 purification Methods 0.000 description 4
- 239000011734 sodium Substances 0.000 description 4
- LPQZERIRKRYGGM-UHFFFAOYSA-N tert-butyl pyrrolidine-1-carboxylate Chemical compound CC(C)(C)OC(=O)N1CCCC1 LPQZERIRKRYGGM-UHFFFAOYSA-N 0.000 description 4
- QAEDZJGFFMLHHQ-UHFFFAOYSA-N trifluoroacetic anhydride Chemical compound FC(F)(F)C(=O)OC(=O)C(F)(F)F QAEDZJGFFMLHHQ-UHFFFAOYSA-N 0.000 description 4
- UHOVQNZJYSORNB-UHFFFAOYSA-N Benzene Chemical compound C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 description 3
- 102000006275 Ubiquitin-Protein Ligases Human genes 0.000 description 3
- 108010083111 Ubiquitin-Protein Ligases Proteins 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 3
- JFDZBHWFFUWGJE-UHFFFAOYSA-N benzonitrile Chemical compound N#CC1=CC=CC=C1 JFDZBHWFFUWGJE-UHFFFAOYSA-N 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 229960004316 cisplatin Drugs 0.000 description 3
- 238000000605 extraction Methods 0.000 description 3
- 239000001963 growth medium Substances 0.000 description 3
- RAXXELZNTBOGNW-UHFFFAOYSA-N imidazole Natural products C1=CNC=N1 RAXXELZNTBOGNW-UHFFFAOYSA-N 0.000 description 3
- 239000012074 organic phase Substances 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- NWZSZGALRFJKBT-KNIFDHDWSA-N (2s)-2,6-diaminohexanoic acid;(2s)-2-hydroxybutanedioic acid Chemical compound OC(=O)[C@@H](O)CC(O)=O.NCCCC[C@H](N)C(O)=O NWZSZGALRFJKBT-KNIFDHDWSA-N 0.000 description 2
- CSCPPACGZOOCGX-MICDWDOJSA-N 1-deuteriopropan-2-one Chemical compound [2H]CC(C)=O CSCPPACGZOOCGX-MICDWDOJSA-N 0.000 description 2
- VHYFNPMBLIVWCW-UHFFFAOYSA-N 4-Dimethylaminopyridine Chemical compound CN(C)C1=CC=NC=C1 VHYFNPMBLIVWCW-UHFFFAOYSA-N 0.000 description 2
- QMHIMXFNBOYPND-UHFFFAOYSA-N 4-methylthiazole Chemical compound CC1=CSC=N1 QMHIMXFNBOYPND-UHFFFAOYSA-N 0.000 description 2
- 102000001301 EGF receptor Human genes 0.000 description 2
- 108060006698 EGF receptor Proteins 0.000 description 2
- 101710113864 Heat shock protein 90 Proteins 0.000 description 2
- 108010006519 Molecular Chaperones Proteins 0.000 description 2
- 206010028980 Neoplasm Diseases 0.000 description 2
- 108091007744 Programmed cell death receptors Proteins 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- 102000013530 TOR Serine-Threonine Kinases Human genes 0.000 description 2
- 108010065917 TOR Serine-Threonine Kinases Proteins 0.000 description 2
- 108010073929 Vascular Endothelial Growth Factor A Proteins 0.000 description 2
- 102000005789 Vascular Endothelial Growth Factors Human genes 0.000 description 2
- 108010019530 Vascular Endothelial Growth Factors Proteins 0.000 description 2
- 230000001028 anti-proliverative effect Effects 0.000 description 2
- 229910052786 argon Inorganic materials 0.000 description 2
- 230000004663 cell proliferation Effects 0.000 description 2
- 239000012043 crude product Substances 0.000 description 2
- 230000003013 cytotoxicity Effects 0.000 description 2
- 231100000135 cytotoxicity Toxicity 0.000 description 2
- 238000001378 electrochemiluminescence detection Methods 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 238000003119 immunoblot Methods 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 239000003112 inhibitor Substances 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- OKKJLVBELUTLKV-VMNATFBRSA-N methanol-d1 Chemical compound [2H]OC OKKJLVBELUTLKV-VMNATFBRSA-N 0.000 description 2
- 230000035772 mutation Effects 0.000 description 2
- VMGAPWLDMVPYIA-HIDZBRGKSA-N n'-amino-n-iminomethanimidamide Chemical compound N\N=C\N=N VMGAPWLDMVPYIA-HIDZBRGKSA-N 0.000 description 2
- SCVFZCLFOSHCOH-UHFFFAOYSA-M potassium acetate Chemical compound [K+].CC([O-])=O SCVFZCLFOSHCOH-UHFFFAOYSA-M 0.000 description 2
- BOLDJAUMGUJJKM-LSDHHAIUSA-N renifolin D Natural products CC(=C)[C@@H]1Cc2c(O)c(O)ccc2[C@H]1CC(=O)c3ccc(O)cc3O BOLDJAUMGUJJKM-LSDHHAIUSA-N 0.000 description 2
- 150000003384 small molecules Chemical class 0.000 description 2
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 2
- VSIVTUIKYVGDCX-UHFFFAOYSA-M sodium;4-[2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)tetrazol-2-ium-5-yl]benzene-1,3-disulfonate Chemical compound [Na+].COC1=CC([N+]([O-])=O)=CC=C1[N+]1=NC(C=2C(=CC(=CC=2)S([O-])(=O)=O)S([O-])(=O)=O)=NN1C1=CC=C([N+]([O-])=O)C=C1 VSIVTUIKYVGDCX-UHFFFAOYSA-M 0.000 description 2
- 239000000758 substrate Substances 0.000 description 2
- 238000000967 suction filtration Methods 0.000 description 2
- 238000001356 surgical procedure Methods 0.000 description 2
- 230000004083 survival effect Effects 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 210000001519 tissue Anatomy 0.000 description 2
- 231100000331 toxic Toxicity 0.000 description 2
- 230000002588 toxic effect Effects 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 239000012224 working solution Substances 0.000 description 2
- SOZMSEPDYJGBEK-LURJTMIESA-N (1s)-1-(4-bromophenyl)ethanamine Chemical compound C[C@H](N)C1=CC=C(Br)C=C1 SOZMSEPDYJGBEK-LURJTMIESA-N 0.000 description 1
- LRFZIPCTFBPFLX-SSDOTTSWSA-N (2s)-3,3-dimethyl-2-[(2-methylpropan-2-yl)oxycarbonylamino]butanoic acid Chemical compound CC(C)(C)OC(=O)N[C@H](C(O)=O)C(C)(C)C LRFZIPCTFBPFLX-SSDOTTSWSA-N 0.000 description 1
- BENKAPCDIOILGV-RQJHMYQMSA-N (2s,4r)-4-hydroxy-1-[(2-methylpropan-2-yl)oxycarbonyl]pyrrolidine-2-carboxylic acid Chemical compound CC(C)(C)OC(=O)N1C[C@H](O)C[C@H]1C(O)=O BENKAPCDIOILGV-RQJHMYQMSA-N 0.000 description 1
- KZPYGQFFRCFCPP-UHFFFAOYSA-N 1,1'-bis(diphenylphosphino)ferrocene Chemical compound [Fe+2].C1=CC=C[C-]1P(C=1C=CC=CC=1)C1=CC=CC=C1.C1=CC=C[C-]1P(C=1C=CC=CC=1)C1=CC=CC=C1 KZPYGQFFRCFCPP-UHFFFAOYSA-N 0.000 description 1
- LMDZBCPBFSXMTL-UHFFFAOYSA-N 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide Substances CCN=C=NCCCN(C)C LMDZBCPBFSXMTL-UHFFFAOYSA-N 0.000 description 1
- VRPJIFMKZZEXLR-UHFFFAOYSA-N 2-[(2-methylpropan-2-yl)oxycarbonylamino]acetic acid Chemical compound CC(C)(C)OC(=O)NCC(O)=O VRPJIFMKZZEXLR-UHFFFAOYSA-N 0.000 description 1
- WGABOZPQOOZAOI-UHFFFAOYSA-N 2-[4-[[(3,5-dimethoxy-4-methylbenzoyl)-(3-phenylpropyl)amino]methyl]phenyl]acetic acid Chemical compound COC1=C(C)C(OC)=CC(C(=O)N(CCCC=2C=CC=CC=2)CC=2C=CC(CC(O)=O)=CC=2)=C1 WGABOZPQOOZAOI-UHFFFAOYSA-N 0.000 description 1
- MNNDREXLRLDWEY-UHFFFAOYSA-N 2-bromo-4-fluorobenzonitrile Chemical compound FC1=CC=C(C#N)C(Br)=C1 MNNDREXLRLDWEY-UHFFFAOYSA-N 0.000 description 1
- FPQQSJJWHUJYPU-UHFFFAOYSA-N 3-(dimethylamino)propyliminomethylidene-ethylazanium;chloride Chemical compound Cl.CCN=C=NCCCN(C)C FPQQSJJWHUJYPU-UHFFFAOYSA-N 0.000 description 1
- VLCARYCTBKNSFG-UHFFFAOYSA-N 4-[[2-(2,6-dioxopiperidin-3-yl)-1-oxo-3H-isoindol-4-yl]amino]butanoic acid Chemical compound O=C1NC(CCC1N1C(C2=CC=CC(=C2C1)NCCCC(=O)O)=O)=O VLCARYCTBKNSFG-UHFFFAOYSA-N 0.000 description 1
- IMLXLGZJLAOKJN-UHFFFAOYSA-N 4-aminocyclohexan-1-ol Chemical compound NC1CCC(O)CC1 IMLXLGZJLAOKJN-UHFFFAOYSA-N 0.000 description 1
- 229960000549 4-dimethylaminophenol Drugs 0.000 description 1
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 1
- DHMQDGOQFOQNFH-UHFFFAOYSA-M Aminoacetate Chemical compound NCC([O-])=O DHMQDGOQFOQNFH-UHFFFAOYSA-M 0.000 description 1
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonia chloride Chemical class [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 description 1
- 238000009020 BCA Protein Assay Kit Methods 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- UHDGCWIWMRVCDJ-CCXZUQQUSA-N Cytarabine Chemical compound O=C1N=C(N)C=CN1[C@H]1[C@@H](O)[C@H](O)[C@@H](CO)O1 UHDGCWIWMRVCDJ-CCXZUQQUSA-N 0.000 description 1
- 101710088194 Dehydrogenase Proteins 0.000 description 1
- 102000001477 Deubiquitinating Enzymes Human genes 0.000 description 1
- 108010093668 Deubiquitinating Enzymes Proteins 0.000 description 1
- 206010059866 Drug resistance Diseases 0.000 description 1
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 1
- 108091006149 Electron carriers Proteins 0.000 description 1
- 102100031181 Glyceraldehyde-3-phosphate dehydrogenase Human genes 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- 102000005431 Molecular Chaperones Human genes 0.000 description 1
- MHABMANUFPZXEB-UHFFFAOYSA-N O-demethyl-aloesaponarin I Natural products O=C1C2=CC=CC(O)=C2C(=O)C2=C1C=C(O)C(C(O)=O)=C2C MHABMANUFPZXEB-UHFFFAOYSA-N 0.000 description 1
- 235000005311 Pandanus odoratissimus Nutrition 0.000 description 1
- 240000002390 Pandanus odoratissimus Species 0.000 description 1
- 239000012083 RIPA buffer Substances 0.000 description 1
- CKNCMXJRFAUMEO-UHFFFAOYSA-N S(=O)(=O)(OC)OC.COC1=CC=CC2=[N+](C3=CC=CC=C3N=C12)C Chemical compound S(=O)(=O)(OC)OC.COC1=CC=CC2=[N+](C3=CC=CC=C3N=C12)C CKNCMXJRFAUMEO-UHFFFAOYSA-N 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000002776 aggregation Effects 0.000 description 1
- 238000004220 aggregation Methods 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 229940098773 bovine serum albumin Drugs 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 230000009702 cancer cell proliferation Effects 0.000 description 1
- 230000005754 cellular signaling Effects 0.000 description 1
- 230000007541 cellular toxicity Effects 0.000 description 1
- 238000002512 chemotherapy Methods 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 238000011281 clinical therapy Methods 0.000 description 1
- 229940126212 compound 17a Drugs 0.000 description 1
- 230000008878 coupling Effects 0.000 description 1
- 238000010168 coupling process Methods 0.000 description 1
- 238000005859 coupling reaction Methods 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- MFXJVHXNGXYJGH-UHFFFAOYSA-N cyclohexyl 2-aminoacetate Chemical compound NCC(=O)OC1CCCCC1 MFXJVHXNGXYJGH-UHFFFAOYSA-N 0.000 description 1
- 229960000684 cytarabine Drugs 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- BADXJIPKFRBFOT-UHFFFAOYSA-N dimedone Chemical compound CC1(C)CC(=O)CC(=O)C1 BADXJIPKFRBFOT-UHFFFAOYSA-N 0.000 description 1
- GPAYUJZHTULNBE-UHFFFAOYSA-N diphenylphosphine Chemical compound C=1C=CC=CC=1PC1=CC=CC=C1 GPAYUJZHTULNBE-UHFFFAOYSA-N 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 238000009509 drug development Methods 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 description 1
- KTWOOEGAPBSYNW-UHFFFAOYSA-N ferrocene Chemical compound [Fe+2].C=1C=C[CH-]C=1.C=1C=C[CH-]C=1 KTWOOEGAPBSYNW-UHFFFAOYSA-N 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 108020004445 glyceraldehyde-3-phosphate dehydrogenase Proteins 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- IKDUDTNKRLTJSI-UHFFFAOYSA-N hydrazine monohydrate Substances O.NN IKDUDTNKRLTJSI-UHFFFAOYSA-N 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 239000012160 loading buffer Substances 0.000 description 1
- 231100000053 low toxicity Toxicity 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 230000001394 metastastic effect Effects 0.000 description 1
- 206010061289 metastatic neoplasm Diseases 0.000 description 1
- 210000003470 mitochondria Anatomy 0.000 description 1
- 239000012046 mixed solvent Substances 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 125000000636 p-nitrophenyl group Chemical group [H]C1=C([H])C(=C([H])C([H])=C1*)[N+]([O-])=O 0.000 description 1
- YJVFFLUZDVXJQI-UHFFFAOYSA-L palladium(ii) acetate Chemical compound [Pd+2].CC([O-])=O.CC([O-])=O YJVFFLUZDVXJQI-UHFFFAOYSA-L 0.000 description 1
- 229960002566 papillomavirus vaccine Drugs 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 235000011056 potassium acetate Nutrition 0.000 description 1
- 230000001376 precipitating effect Effects 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 229940121649 protein inhibitor Drugs 0.000 description 1
- 239000012268 protein inhibitor Substances 0.000 description 1
- 238000011865 proteolysis targeting chimera technique Methods 0.000 description 1
- 238000010791 quenching Methods 0.000 description 1
- 238000001959 radiotherapy Methods 0.000 description 1
- 230000000306 recurrent effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000010898 silica gel chromatography Methods 0.000 description 1
- 235000020183 skimmed milk Nutrition 0.000 description 1
- 108010026668 snake venom protein C activator Proteins 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- HJEZRYIJNHAIGY-UHFFFAOYSA-N tert-butyl 4-bromobutanoate Chemical compound CC(C)(C)OC(=O)CCCBr HJEZRYIJNHAIGY-UHFFFAOYSA-N 0.000 description 1
- PSELCIMQRLODQE-UHFFFAOYSA-N tert-butyl 6-bromohexanoate Chemical compound CC(C)(C)OC(=O)CCCCCBr PSELCIMQRLODQE-UHFFFAOYSA-N 0.000 description 1
- PKNFPFFOAWITLF-RZUBCFFCSA-N tert-butyl N-[(2S)-1-[(2S,4R)-4-hydroxy-2-[[4-(4-methyl-1,3-thiazol-5-yl)phenyl]methylcarbamoyl]pyrrolidin-1-yl]-3,3-dimethyl-1-oxobutan-2-yl]carbamate Chemical compound Cc1ncsc1-c1ccc(CNC(=O)[C@@H]2C[C@@H](O)CN2C(=O)[C@@H](NC(=O)OC(C)(C)C)C(C)(C)C)cc1 PKNFPFFOAWITLF-RZUBCFFCSA-N 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 238000010798 ubiquitination Methods 0.000 description 1
- 230000034512 ubiquitination Effects 0.000 description 1
- 238000001291 vacuum drying Methods 0.000 description 1
- 239000012130 whole-cell lysate Substances 0.000 description 1
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
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
- A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
- A61K31/445—Non condensed piperidines, e.g. piperocaine
- A61K31/4523—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems
- A61K31/454—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems containing a five-membered ring with nitrogen as a ring hetero atom, e.g. pimozide, domperidone
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P15/00—Drugs for genital or sexual disorders; Contraceptives
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- 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
- C07D401/14—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 three or more hetero rings
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
Landscapes
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Veterinary Medicine (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Endocrinology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Engineering & Computer Science (AREA)
- Reproductive Health (AREA)
- Epidemiology (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- Genetics & Genomics (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
本发明公开了一种Hsp90PROTAC化合物及其制备方法与在制备抗宫颈癌药物中的应用,属于医药技术领域。本发明的Hsp90PROTAC化合物的结构如通式(I)所示,Hsp90抑制剂SNX‑5422分别与带有不同连接子的来那度胺羧酸衍生物和VHL羧酸衍生物为原料在缩合剂HATU和碱DIPEA的催化下,通过一步酰胺反应制备得到Hsp90PROTAC化合物。本发明的Hsp90PROTAC化合物有很好抗宫颈癌活性,其中部分化合物的抑制活性达到了纳摩尔水平,其可用于制备抗宫颈癌药物。
Description
技术领域
本发明属于医药技术领域,具体涉及一种靶向降解Hsp90的PROTACs化合物及其制备方法与在制备抗宫颈癌药物中的应用。
背景技术
宫颈癌是一种在女性群体当中高发的恶性肿瘤,也是导致女性死亡的主要肿瘤之一,对女性的生命健康构成严重威胁。尽管过去几十年人们一直致力于宫颈癌预防和治疗策略的开发,特别是HPV疫苗的上市在一定程度上降低了发病率,但每年仍有相当一部分女性罹患宫颈癌;治疗手段方面,目前临床上鲜有针对宫颈癌的特异性治疗药物,手术和放化疗仍旧是主要的临床疗法。然而手术治疗大多适用于早期宫颈癌的治疗,对于转移性和复发的宫颈癌患者的治疗效果十分有限,后者的五年生存率不足20%;而阿糖胞苷、顺铂等非选择性的化疗药物往往具有很强的毒副作用,治疗效果也十分有限。因此,为了改善宫颈癌临床治疗效果,同时降低治疗药物的脱靶毒副作用,靶向治疗药物的研发显得尤为重要。
目前,宫颈癌靶向药物研究多数集中在表皮生长因子受体(EGFR)、血管内皮生长因子(VEGF)、哺乳动物雷帕霉素靶蛋白(mTOR)以及程序性死亡受体(PD-1)等。尽管目前已有宫颈癌相关的抗体治疗药物上市,但总体来看宫颈癌的靶向药物研究仍旧处于早期阶段,尤其是小分子治疗药物的开发。热休克蛋白90(Hsp90)作为一种分子伴侣蛋白,其主要功能是保证底物蛋白进行正确折叠并防止其非特异性聚集,以维持细胞存活、增殖和分化。该伴侣分子的底物蛋白十分广泛,这使得它与肿瘤的发生发展有着密切关系。研究发现,Hsp90蛋白在宫颈癌组织中的表达异常增高,其在宫颈癌的发生发展过程中可能起着重要作用。此外,部分已报道的Hsp90抑制剂对宫颈癌表现出了一定的抑制活性,这进一步表明其可作为宫颈癌靶向治疗药物开发的潜在靶点。然而,同大多数靶蛋白抑制剂一样,Hsp90抑制剂用于宫颈癌治疗同样将面临耐药突变带来的严峻挑战。
蛋白水解靶向嵌合体(PROTAC)技术是一种基于泛素-蛋白酶体系统发展起来的新兴药物开发策略,该技术通过构建一个由兴趣蛋白配体(POI ligand)、连接子(Linker)和E3泛素连接酶配体(E3 ligand)偶联形成的异源双功能分子,诱导目标蛋白和E3连接酶在空间上相互靠近,使得前者被泛素化标记和降解,从而清除致病蛋白,发挥治疗疾病的作用。该技术相较于传统的小分子抑制剂具有很大的优势,例如其对结合亲和力要求较低,这使得它能够靶向不可成药蛋白、克服靶点耐药突变。此外,它还具有用量小、活性高、选择性高和低毒性等特点。鉴于此,将PROTAC技术用于开发一系列靶向降解Hsp90的PROTAC分子,将是一种具有潜力宫颈癌的靶向治疗策略。
发明内容
本发明的目的在于克服现有技术存在的不足,提供一种Hsp90 PROTAC化合物及其应用,所述的Hsp90 PROTAC化合物具有显著的抗宫颈癌活性,可以作为新的抗宫颈癌药物进行开发,具有广泛的应用前景。本发明的目的还在于提供所述的Hsp90 PROTAC化合物的制备方法。
为了实现上述目的,本发明所采取的技术方案如下:
第一方面,本发明提供了一种通式(I)所示的Hsp90 PROTAC化合物或其药理或生理上可接受的盐,
其中,
Linker选自
E3 ligand选自
本发明通过体外抗宫颈癌活性测试发现,上述Hsp90 PROTAC化合物能够以浓度依赖的方式降解Hsp90,同时对宫颈癌细胞具有显著的抗增殖活性,可用于制备Hsp90降解剂、抗宫颈癌药物。
优选地,所述的Hsp90 PROTAC化合物选自如下表1所示的化合物:
表1
第二方面,本发明提供了上述任意一种Hsp90 PROTAC化合物或其药理或生理上可接受的盐在制备Hsp90降解剂或抗宫颈癌药物中的应用。
第三方面,本发明提供了提供一种Hsp90降解剂或抗宫颈癌的药物,包含上述Hsp90PROTAC化合物或其药理或生理上可接受的盐中的一种或多种。所述的药物还可包含药学上可接受的载体或赋形剂,其可按照现有的常规医药技术来制备。
第四方面,本发明提供了上述Hsp90 PROTAC化合物的制备方法。所述的Hsp90PROTAC化合物的制备方法包括如下步骤:采用Hsp90抑制剂SNX-5422与E3连接酶配体(来那度胺或VHL)的羧酸衍生物在缩合剂HATU和碱EIPEA的作用下进行酰胺缩合反应得到所述Hsp90 PROTAC化合物:
Hsp90 PROTAC化合物的合成路线
优选的,所述的Hsp90 PROTAC化合物的制备方法中,SNX-5422、VHL羧酸衍生物、HATU和DIPEA投料的摩尔比为1:1.0~1.2:1.0~1.2:2.5~3.5;SNX-5422、来那度胺羧酸衍生物、HATU和DIPEA投料的摩尔比为1:1.0~1.2:1.0~1.2:2.5~3.5。
本发明的优点和有益效果:本发明的Hsp90 PROTAC化合物有很好抗宫颈癌活性,其中部分化合物的抑制活性达到了纳摩尔水平。本发明的Hsp90 PROTAC化合物可作为新的抗宫颈癌药物进行开发,具有广泛的应用前景。
附图说明
图1是免疫印迹分析PROTAC化合物对Hsp90的体外降解活性。A是1μM的Hsp90PROTAC化合物对Hsp90的体外降解活性;B是不同浓度(μM)LF8对Hsp90的体外降解活性,其中,SNX-5422浓度为1μM。
具体实施方式
下面通过实施例对本发明做进一步详细的描述。所提供的实施例仅是对本发明的说明,而不以任何方式限制本发明揭示的其余内容。
本发明通式(I)所示的Hsp90 PROTAC化合物的制备具体包括以下步骤:
1、通过下式(1)所示反应合成得到6,6-二甲基-3-(三氟甲基)-1,5,6,7-四氢-4H-吲唑-4-酮(2),并作为下一步反应的原料。
首先,取一100mL圆底烧瓶并加入THF(20mL),随后,冰浴条件下依次加入咪唑(imidazole)(6799mg,99.87mmol)和三氟乙酸酐(TFAA)(8mL),搅拌30min。另取一20mL圆底烧瓶加入THF(10mL),并依次加入市售5,5-二甲基-1,3-环己二酮1(4000mg,28.53mmol)和imidazole(2914mg,42.80mmol),室温条件下搅拌至所有固体完全溶解,并将其缓慢滴加到上述反应液中,滴加完毕,继续搅拌5h。TLC监测,待原料1完全消失后,利用真空旋转蒸发仪除去反应液中的THF,并将剩余物溶于DCM,利用DCM进行萃取,合并有机相后分别用1M HCl和饱和氯化钠进行洗涤,无水硫酸钠进行干燥,旋干。将其重新溶解于EtOH(12mL),冰浴条件下,缓慢滴加水合肼(NH2NH2·H2O,6mL),加料完毕后,继续搅拌1h,TLC监测,原料消失后,往反应体系中缓慢滴加饱和氯化钠,有淡黄色固体析出,抽滤,真空干燥即得淡黄色固体产物2(4307mg,18.55mmol)。
2、通过下式(2)所示反应合成得到2-溴-4-(6,6-二甲基-4-氧代-3-(三氟甲基)-4,5,6,7-四氢-1H-吲唑-1-基)苄腈(4),并作为下一步反应的原料。
将反应(1)制得的原料2(2000mg,8.61mmol)溶于DMSO(6mL),并依次加入市售2-溴-4-氟苯甲腈3(1723mg,8.61mmol)和DIPEA(3352mg,25.84mmol),将反应体系置于油浴锅中加热至85℃,搅拌3h。TLC监测,原料完全消失后,利用DCM对反应体系进行萃取,饱和氯化钠洗涤,无水硫酸钠进行干燥,旋干后即得粗产品,利用柱层析进行纯化,流动相为石油醚/乙酸乙酯(V/V=3/1),得淡黄色固体产物4(2947mg,7.15mmol)。
3、通过下式(3)所示反应合成得到4-(6,6-二甲基-4-氧代-3-(三氟甲基)-4,5,6,7-四氢-1H-吲唑-1-基)-2-(((1r,4r)-4-羟基环己基)氨基)苄腈(6),并作为下一步反应的原料。
将反应(2)制得的原料4(1280mg,3.11mmol)、市售反式-4-氨基环己醇5(1073mg,9.32mmol)、Pd(OAc)2(35mg,0.16mmol)、1,1'-双(二苯基膦)二茂铁(DPPF)(172mg,0.31mmol)和Cs2CO3(3035mg,9.32mmol)依次加入50mL封管中,随后通入氩气并加入甲苯(Toluene)(18mL),将其置于微波反应器中,120℃下反应3h。TLC监测,原料消失后,利用硅藻土进行抽滤,将滤液浓缩后进行柱层析纯化,流动相为石油醚/乙酸乙酯(V/V=1/1-1/3),得淡黄色固体产物6(984mg,2.20mmol)。
4、通过下式(4)所示反应合成得到4-(6,6-二甲基-4-氧代-3-(三氟甲基)-4,5,6,7-四氢-1H-吲唑-1-基)-2-(((1r,4r)-4-羟基环己基)氨基)苯甲酰胺(7),并作为下一步反应的原料。
取一50mL单口烧瓶并加入EtOH(10mL),随后加入反应(3)制得的原料6(900mg,2.02mmol)和DMSO(2mL),室温下搅拌至固体完全溶解,接着依次缓慢滴加1N NaOH溶液(2mL)和质量分数为30%的H2O2溶液(4mL),加料完毕,继续搅拌2h。TLC监测,原料完全消失后,缓慢滴加饱和氯化铵溶液淬灭反应,5分钟后缓慢滴加饱和氯化钠溶液至有白色固体析出,抽滤,真空干燥后即得白色固体产物7(852mg,1.83mmol)。
5、通过下式(5)所示反应合成得到(1r,4r)-4-((2-氨基-5-(6,6-二甲基-4-氧代-3-(三氟甲基)-4,5,6,7-四氢-1H-吲唑-1-基)苯基)氨基)甘氨酸环己基酯(SNX-5422),并作为下一步反应的原料。
将反应(4)制得的原料7(800mg,1.72mmol)、BOC-甘氨酸(453mg,2.58mmol)、DMAP(316mg,2.58mmol)和EDCI(660mg,3.44mmol)加入50mL单口烧瓶,并加入无水DCM(10mL),室温下搅拌过夜。TLC监测,原料完全消失后,利用DCM对反应体系进行萃取,无水硫酸钠进行干燥,旋干后将其重新溶于DCM(6mL),冰浴条件下缓慢滴加TFA(2mL),继续搅拌1.5h。TLC进行监测,原料消失后,利用真空旋转蒸发仪除去DCM和TFA后,溶于DCM并利用氨水调节体系的pH>7,DCM萃取,旋干,利用柱层析进行纯化,流动相为ECM/MeOH(V/V=15/1),得到白色固体产物SNX-5422(668mg,1.28mmol)。
6、通过下式(6)所示反应合成得到(4-溴苄基)氨基甲酸叔丁酯(9),并作为下一步反应的原料。
取一100mL单口烧瓶,并加入混合溶剂EtOAc/H2O(V/V=1/1)(20mL),随后依次加入(S)-(-)-1-(4-溴苯)乙胺8(6000mg,29.99mmol)和NaHCO3(2771mg,32.99mmol),冰浴条件下缓慢加入(Boc)2O(7199mg,32.99mmol),加料完毕,撤去冰浴,室温下搅拌3h。TLC监测反应,原料8完全消失后,利用EtOAc对反应体系进行萃取,饱和氯化钠洗涤,无水硫酸钠干燥,旋干即得粗产品,利用粗层析进行纯化,流动相为石油醚/乙酸乙酯(V/V=15:1),得白色固体产物9(8732mg,29.09mmol)。
7、通过下式(7)所示反应合成得到(4-(4-甲基噻唑-5-基)苄基)氨基甲酸叔丁酯(11),并作为下一步反应的原料。
将反应(6)制得的原料9(8500mg,28.31mmol)、市售4-甲基噻唑10(5615mg,56.63mmol)、醋酸钯(321mg,1.42mmol)、醋酸钾(11115mg,113.26mmol)置于50mL圆底烧瓶中,抽真空后通入氩气,循环三次,随后加入溶剂DMAC,150℃下反应过夜。TLC监测反应,原料9完全消失后,加水淬灭反应,乙酸乙酯萃取。有机相浓缩后,使用流动相为石油醚/乙酸乙酯(V/V=6:1)经硅胶柱层析分离纯化,得到白色固体产物11(7303mg,22.93mmol)。
8、通过下式(8)所示反应合成得到(2S,4R)-4-羟基-2-(((4-(4-甲基噻唑-5-基)苄基)氨基甲酰基)吡咯烷-1-羧酸叔丁酯(12),并作为下一步反应的原料。
取一100mL单口烧瓶并加入DCM(20mL),然后加入反应(7)制得的原料11(7000mg,21.98mmol),冰浴条件下,缓慢滴加TFA(10026mg,87.93mmol),加料完毕,继续搅拌1h。TLC监测反应,原料消失后旋蒸除去体系中的DCM和TFA,并重新将剩余物溶于DCM,利用氨水调节pH至碱性,DCM萃取,旋蒸除去大部分DCM后,补加适量无水DCM(30mL),然后依次加入DIPEA(11408mg,87.93mmol)、HATU(9194mg,24.18mmol)和Boc-Hyp-OH(5592mg,24.18mmol),加料完毕后,室温下搅拌3h。TLC监测,原料完全消失后,利用DCM进行萃取,旋干后进行柱层析纯化,流动相为石油醚/乙酸乙酯(V/V=3/1),得白色固体产物12(8021mg,18.59mmol)。
9、通过下式(9)所示反应合成得到叔丁基((S)-1-((2S,4R)-4-羟基-2-((4-(4-甲基噻唑-5-基)苄基)氨基甲酰基)吡咯烷-1-基)-3,3-二甲基-1-氧代丁烷-2-基)氨基甲酸酯(Boc-VHL),并作为下一步反应的原料。
取一50mL单口烧瓶并加入DCM(50mL),然后加入反应(8)制得的原料12(7800mg,18.07mmol),冰浴条件下,缓慢滴加TFA(8244mg,72.30mmol),加料完毕,继续搅拌1h。TLC监测反应,原料消失后旋蒸除去体系中的DCM和TFA,并重新将剩余物溶于适量DCM,利用氨水调节pH至碱性,DCM萃取,干燥,旋蒸除去大部分DCM后,补加适量无水DCM(30mL),然后依次加入DIPEA(93.80mg,72.30mmol)、HATU(7560mg,19.88mmol)和Boc-Tle-OH(4598mg,19.88mmol),加料完毕后,室温下搅拌3h。TLC监测,原料完全消失后,利用DCM进行萃取,旋干后进行柱层析纯化,流动相为二氯甲烷/甲醇(V/V=30:1),得白色固体产物Boc-VHL(8057mg,14.79mmol)。
10、通过下式(10)、(11)所示反应合成得到含不同长度Linker的VHL羧酸衍生物(15a-c),并作为下一步反应的原料。
以化合物6-(((S)-1-((2S,4R)-4-羟基-2-(((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)氨基甲酰基)吡咯烷-1-基)-3,3-二甲基-1-氧代丁-2-基)氨基)己酸15a的制备为例,具体步骤如下:取一50mL单口烧瓶并加入DCM,然后加入反应(9)制得的原料Boc-VHL(400mg,0.73mmol),冰浴条件下,缓慢滴加TFA(335mg,2.94mmol),加料完毕,继续搅拌1h。TLC监测反应,原料消失后旋蒸除去体系中的DCM和TFA,并重新将剩余物溶于适量DCM,利用氨水调节pH至碱性,DCM萃取,干燥后旋蒸除去DCM,随后将其溶于MeCN(6mL),依次加入6-溴己酸叔丁酯13a(221mg,0.88mmol)、KI(6mg,0.04mmol)和K2CO3(304mg,2.20mmol),加料完毕后将反应置于油浴锅,并加热至85℃,搅拌过夜。TLC监测反应,原料消失后利用DCM对反应进行萃取,干燥后旋干有机相。将剩余物重新溶于DCM(3mL),冰浴条件下,缓慢滴加TFA(1mL),滴加完毕后,撤去冰浴并继续搅拌1.5h。TLC监测反应,原料消失后旋蒸除去体系中的DCM和TFA,并将剩余物溶于乙酸乙酯,乙酸乙酯萃取、干燥,旋蒸浓缩后即得白色固体产物15a(275mg,0.49mmol)。
化合物15b和15c的制备方法同上。
11、通过下式(12)所示反应合成得到系列(A)Hsp90 PROTAC化合物(LF1-LF3)。
以化合物(1R,4S)-4-((2-氨基甲酰基-5-(6,6-二甲基-4-氧代-3-(三氟甲基)-4,5,6,7-四氢-1H-吲唑-1-基)苯基)氨基)环己基(6-(((S)-1-((2S,4R)-4-羟基-2-((((S)-1-(4-(4-甲基噻唑-5-基)苯)乙基)氨基甲酰基)吡咯烷-1-基)-3,3-二甲基-1-氧代丁-2-基)氨基)己酰基)甘氨酸酯LF1的制备为例,步骤如下:称取化合物SNX-5422(100mg,0.19mmol)、15a(118mg,0.21mmol)、HATU(80mg,0.21mmol)、DIPEA(75mg,0.58mmol)置于25mL圆底烧瓶中,加入无水DCM,室温条件下搅拌过夜,TLC监测原料SNX-5422完全消失后,利用DCM进行萃取,无水硫酸钠进行干燥,浓缩后进行柱层析纯化,流动相为二氯甲烷/甲醇(V/V=15:1),得白色固体产物LF1(167mg 0.16mmol)。
Hsp90 PROTAC化合物LF2和LF3的制备方法同上。
12、通过下式(13)所示反应合成得到含不同长度Linker的来那度胺羧酸衍生物(17a-f),并作为下一步反应的原料。
以化合物4-((2-(2,6-二氧哌啶-3-基)-1-氧代异吲哚啉-4-基)氨基)丁酸17a的制备为例,步骤如下:取一25mL单口烧瓶,将来那度胺(800mg,3.09mmol)、市售4-溴丁酸叔丁酯16a(826mg,3.70mmol)和DIPEA(1201mg,9.26mmol)依次溶于NMP(6mL),然后将反应体系置于油浴锅,加热至110℃,搅拌过夜。TLC监测,来那度胺完全消失后,将反应冷却至室温,并缓慢滴加饱和氯化钠溶液,有淡棕色固体析出,抽滤并烘干固体,然后将其溶于DCM(6mL),冰浴条件下缓慢滴加TFA(6mL),滴加完毕,撤去冰浴使反应体系温度升至室温,搅拌2h。TLC进行监测,原料消失后,浓缩反应体系并加入适量乙酸乙酯,剧烈搅拌30min,有米白色固体析出,抽滤即得化合物17a(750mg,2.17mmol)。
化合物17b-f的制备方法同上。
13、通过下式(14)所示反应合成得到系列(B)Hsp90 PROTAC化合物(LF4-LF9)。
以化合物(1R,4R)-4-((2-氨基-5-(6,6-二甲基-4-氧代-3-(三氟甲基)-4,5,6,7-四氢-1H-吲唑-1-基)苯基)氨基)环己基(4-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-4-基)氨基)丁酰基)甘氨酸酯LF4的制备为例,步骤如下:称取化合物SNX-5422(100mg,0.19mmol)、17a((73mg,0.21mmol)、HATU(80mg,0.21mmol)、DIPEA(75mg,0.58mmol)置于25mL圆底烧瓶中,加入无水DCM,室温条件下搅拌过夜,TLC监测原料SNX-5422完全消失后,利用DCM进行萃取,无水硫酸钠进行干燥,浓缩后进行柱层析纯化,流动相为二氯甲烷/甲醇(V/V=15:1),得白色固体产物LF4(130mg,0.15mmol)。
Hsp90 PROTAC化合物LF5-LF9的制备方法同上。
【实施例1】(1R,4S)-4-((2-氨基甲酰基-5-(6,6-二甲基-4-氧代-3-(三氟甲基)-4,5,6,7-四氢-1H-吲唑-1-基)苯基)氨基)环己基(6-(((S)-1-((2S,4R)-4-羟基-2-((((S)-1-(4-(4-甲基噻唑-5-基)苯)乙基)氨基甲酰基)吡咯烷-1-基)-3,3-二甲基-1-氧代丁-2-基)氨基)己酰基)甘氨酸酯LF1的制备。
称取化合物SNX-5422(100mg,0.19mmol)、15a((118mg,0.21mmol)、HATU(80mg,0.21mmol)、DIPEA(75mg,0.58mmol)置于25mL圆底烧瓶中,加入无水DCM,室温条件下搅拌过夜,TLC监测原料SNX-5422完全消失后,利用DCM进行萃取,无水硫酸钠进行干燥,浓缩后进行柱层析纯化,流动相为二氯甲烷/甲醇(V/V=15:1),得白色固体产物LF1(167mg,0.16mmol),产率为82%。
1H NMR(400MHz,Chloroform-d)δ8.66(s,1H),8.21(d,J=7.2Hz,1H),7.93(d,J=7.7Hz,1H),7.63(d,J=8.4Hz,1H),7.37(s,4H),6.81-6.72(m,2H),6.58(d,J=8.3Hz,1H),5.05(t,J=7.3Hz,1H),4.83(dt,J=15.2,7.6Hz,2H),4.54(s,1H),3.96(d,J=5.5Hz,2H),3.75(d,J=11.1Hz,1H),3.59(d,J=8.1Hz,1H),3.39(s,1H),3.15(s,1H),2.85(s,2H),2.51(d,J=9.1Hz,4H),2.44(d,J=15.4Hz,3H),2.22(t,J=7.4Hz,2H),2.12–1.95(m,5H),1.64–1.28(m,17H),1.11(s,6H),1.00(s,9H).13C NMR(101MHz,Chloroform-d)δ190.34,174.14,171.48,170.22,169.78,150.73,150.45,150.15,148.30,143.49,141.53,131.74,130.62,129.50,126.44,116.14,113.46,109.16,107.11,73.10,69.86,67.13,58.47,56.49,52.24,49.40,48.87,48.29,41.50,37.13,35.80,35.68,35.27,29.47,29.19,28.22,26.80,26.31,24.97,22.36,16.06.HRMS(ESI)m/z calcd for C54H70F3N9O8SNa(M+Na)+:1084.4918;found,1084.4928.
【实施例2】(1R,4R)-4-((2-氨基甲酰基-5-(6,6-二甲基-4-氧代-3-(三氟甲基)-4,5,6,7-四氢-1H-吲唑-1-基)苯基)氨基)环己基(8-(((S)-1-((2S,4R)-4-羟基-2-((((S)-1-(4-(4-甲基噻唑-5-基)苯)乙基)氨基甲酰基)吡咯烷-1-基)-3,3-二甲基-1-氧代丁-2-基)氨基)辛酰基)甘氨酸酯LF2的制备。
制备方法同上述【实施例1】所述,区别在于将原料15a替换为原料15b,得白色固体产物LF2,产率为83%。
1H NMR(400MHz,Chloroform-d)δ8.67(s,1H),8.21(d,J=7.3Hz,1H),7.95(d,J=7.7Hz,1H),7.61(d,J=8.4Hz,1H),7.38(s,4H),6.78(s,1H),6.59(d,J=8.5Hz,2H),5.06(t,J=7.2Hz,1H),4.85(q,J=8.7,8.1Hz,2H),4.57(s,1H),3.98(d,J=4.4Hz,2H),3.73(d,J=11.1Hz,1H),3.61(dd,J=11.0,4.3Hz,1H),3.40(s,1H),3.21(s,1H),2.85(s,2H),2.51(s,4H),2.47(s,3H),2.22(t,J=7.5Hz,2H),2.17–1.93(m,5H),1.68–1.34(m,13H),1.27(d,J=5.8Hz,8H),1.12(s,6H),1.02(s,9H).13C NMR(101MHz,Chloroform-d)δ190.36,174.10,171.48,170.18,169.74,150.73,150.48,150.14,148.28,143.46,141.53,131.75,130.62,130.29,129.50,126.45,116.14,113.46,109.16,73.05,69.81,67.22,58.50,52.23,49.42,48.88,48.69,37.14,36.00,35.68,35.27,29.44,29.20,28.62,28.22,26.82,26.60,25.30,22.36,16.05.HRMS(ESI)m/z calcd for C56H74F3N9O8SNa(M+Na)+:1112.5231;found,1112.5232.
【实施例3】(1R,4R)-4-((2-氨基甲酰基-5-(6,6-二甲基-4-氧代-3-(三氟甲基)-4,5,6,7-四氢-1H-吲唑-1-基)苯基)氨基)环己基(10-(((S)-1-((2S,4r)-4-羟基-2-((((S)-1-(4-(4-甲基噻唑-5-基)苯)乙基)氨基甲酰基)吡咯烷-1-基)-3,3-二甲基-1-氧代丁-2-基)氨基)癸酰基)甘氨酸酯LF3的制备。
制备方法同上述【实施例1】所述,区别在于将原料15a替换为原料15c,得白色固体产物LF3,产率为88%。
1H NMR(400MHz,Chloroform-d)δ8.64(s,1H),8.19(d,J=7.4Hz,1H),7.91(d,J=7.8Hz,1H),7.58(d,J=8.4Hz,1H),7.35(s,4H),6.73(d,J=2.0Hz,1H),6.56(d,J=8.4Hz,1H),6.42(d,J=8.0Hz,1H),5.02(q,J=6.9Hz,1H),4.82(t,J=7.2Hz,2H),4.55(t,J=4.4Hz,1H),3.96(d,J=5.3Hz,2H),3.62(dt,J=9.9,6.6Hz,2H),3.37(d,J=9.0Hz,1H),3.09(s,1H),2.82(s,2H),2.55(d,J=14.1Hz,1H),2.48(s,3H),2.44(s,2H),2.34–2.29(m,1H),2.19(t,J=7.5Hz,2H),2.08(d,J=12.5Hz,2H),2.01–1.94(m,2H),1.60–1.50(m,4H),1.46–1.37(m,9H),1.28–1.17(m,12H),1.09(s,6H),0.97(s,9H).13C NMR(151MHz,DMSO-d6)δ190.28,172.88,170.83,169.53,152.00,151.50,149.54,147.88,147.79,144.79,140.69,131.18,130.78,129.72,129.02,128.88,126.37,121.55,115.39,114.06,109.13,106.84,72.12,68.81,66.19,58.39,56.25,51.72,48.73,48.11,47.76,40.90,37.51,36.04,35.20,35.08,34.89,29.27,29.09,28.97,28.83,28.62,27.54,26.82,26.61,26.57,25.25,22.52,22.20,16.01.HRMS(ESI)m/z calcd for C58H78F3N9O8SNa(M+Na)+:1140.5544;found,1140.5540.
【实施例4】(1R,4R)-4-((2-氨基-5-(6,6-二甲基-4-氧代-3-(三氟甲基)-4,5,6,7-四氢-1H-吲唑-1-基)苯基)氨基)环己基(4-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-4-基)氨基)丁酰基)甘氨酸酯LF4的制备。
称取化合物SNX-5422(100mg,0.19mmol)、17a((73mg,0.21mmol)、HATU(80mg,0.21mmol)、DIPEA(75mg,0.58mmol)置于25mL圆底烧瓶中,加入无水DCM,室温条件下搅拌过夜,TLC监测原料SNX-5422完全消失后,利用DCM进行萃取,无水硫酸钠进行干燥,浓缩后进行柱层析纯化,流动相为二氯甲烷/甲醇(V/V=15:1),得白色固体产物LF4(130mg,0.15mmol),产率为80%。
1H NMR(400MHz,Acetone-d6)δ9.87(s,1H),8.61(d,J=7.6Hz,1H),7.87(d,J=8.4Hz,1H),7.55(t,J=5.9Hz,2H),7.31(t,J=7.7Hz,1H),7.02(d,J=7.4Hz,1H),6.98(d,J=2.1Hz,1H),6.84(d,J=8.0Hz,1H),6.77(dd,J=8.4,2.1Hz,1H),5.27–5.09(m,2H),4.81(dt,J=9.9,5.3Hz,1H),4.37–4.18(m,2H),3.94(d,J=5.9Hz,2H),3.54(dd,J=11.0,6.8Hz,1H),3.30(q,J=6.4Hz,2H),3.08(s,2H),3.04–2.97(m,1H),2.78–2.71(m,1H),2.51–2.43(m,3H),2.41(t,J=7.0Hz,2H),2.22–2.11(m,3H),2.04–1.92(m,4H),1.59(dt,J=12.8,9.4Hz,2H),1.46(ddt,J=16.0,9.6,4.9Hz,2H),1.13(s,6H).13C NMR(151MHz,DMSO-d6)δ190.23,172.92,172.62,171.26,170.79,169.53,168.91,152.00,149.51,143.66,140.66,138.52,132.06,130.73,129.21,126.51,121.53,119.74,115.35,114.04,111.75,110.00,109.13,106.86,54.91,53.47,51.69,51.51,48.68,45.74,42.17,40.89,35.99,35.17,32.54,31.26,29.28,29.10,27.51,24.55,22.84,18.03,16.71,12.34.HRMS(ESI)m/z calcd for C42H48F3N8O8(M+H)+:849.3547;found,849.3544.
【实施例5】(1R,4R)-4-((2-氨基-5-(6,6-二甲基-4-氧代-3-(三氟甲基)-4,5,6,7-四氢-1H-吲唑-1-基)苯基)氨基)环己基(5-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-4-基)氨基)戊酰基)甘氨酸酯LF5的制备。
制备方法同上述【实施例4】所述,区别在于将原料17a替换为原料17b,得白色固体产物LF5,产率为69%。
1H NMR(400MHz,Acetone-d6)δ9.99(s,1H),8.57(d,J=7.5Hz,1H),7.87(d,J=8.4Hz,1H),7.67(s,1H),7.58(t,J=5.9Hz,1H),7.28(t,J=7.7Hz,1H),7.01(d,J=7.4Hz,1H),6.97(d,J=2.1Hz,1H),6.87(s,1H),6.81–6.75(m,2H),5.19(dd,J=13.3,5.1Hz,1H),5.04(t,J=5.5Hz,1H),4.78(dt,J=9.8,5.3Hz,1H),4.34–4.22(m,2H),3.93(d,J=5.9Hz,2H),3.56–3.47(m,1H),3.23(q,J=6.2Hz,2H),3.06(s,2H),2.91(d,J=5.4Hz,1H),2.73(dt,J=17.2,3.6Hz,1H),2.44(s,3H),2.30(t,J=6.8Hz,2H),2.13(tt,J=15.6,2.6Hz,3H),2.01–1.94(m,2H),1.78–1.65(m,4H),1.60–1.50(m,2H),1.43(td,J=12.4,11.9,6.2Hz,2H),1.11(s,6H).13C NMR(151MHz,DMSO-d6)δ190.23,172.92,172.68,171.26,170.79,169.51,168.92,151.99,149.50,143.74,140.66,138.52,138.26,132.05,130.73,129.20,126.47,121.53,119.74,115.35,114.03,111.75,109.92,109.12,106.85,72.15,53.47,51.69,51.51,48.68,45.76,42.44,41.72,40.84,35.99,35.17,34.75,31.26,29.28,29.08,27.99,27.52,27.50,22.85,22.83,18.03,16.71.HRMS(ESI)m/z calcd forC43H50F3N8O8(M+H)+:863.3704;found,863.3712.
【实施例6】(1R,4R)-4-((2-氨基-5-(6,6-二甲基-4-氧代-3-(三氟甲基)-4,5,6,7-四氢-1H-吲唑-1-基)苯基)氨基)环己基(6-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-4-基)氨基)己酰基)甘氨酸酯LF6的制备。
制备方法同上述【实施例4】所述,区别在于将原料17a替换为原料17c,得白色固体产物LF6,产率为68%。
1H NMR(400MHz,DMSO-d6)δ11.05(s,1H),8.46(d,J=7.6Hz,1H),8.27(t,J=6.0Hz,1H),8.04(s,1H),7.81(d,J=8.4Hz,1H),7.38(s,1H),7.27(t,J=7.7Hz,1H),6.97–6.87(m,2H),6.78–6.69(m,2H),5.59(t,J=5.5Hz,1H),5.13(dd,J=13.3,5.1Hz,1H),4.73(dt,J=10.1,5.6Hz,1H),4.31–4.05(m,2H),3.80(d,J=5.8Hz,2H),3.12(q,J=6.6Hz,2H),2.99(s,2H),2.97–2.85(m,1H),2.68–2.59(m,1H),2.45(s,2H),2.32(qd,J=13.1,4.3Hz,1H),2.16(t,J=7.3Hz,2H),2.09–1.99(m,3H),1.97–1.87(m,2H),1.57(tq,J=19.9,11.6,9.3Hz,6H),1.44–1.31(m,4H),1.04(s,6H).13C NMR(101MHz,DMSO-d6)δ190.77,173.45,173.26,171.75,171.27,170.00,169.43,152.46,149.95,144.20,141.11,132.47,131.20,129.70,126.90,115.79,114.44,112.15,110.38,109.58,107.28,72.60,52.12,51.94,49.13,46.19,43.07,41.30,36.44,35.63,35.46,31.71,29.72,29.54,28.71,27.94,26.66,25.50,23.27.HRMS(ESI)m/z calcd for C44H52F3N8O8(M+H)+:877.3860;found,877.3854.
【实施例7】(1R,4R)-4-((2-氨基-5-(6,6-二甲基-4-氧代-3-(三氟甲基)-4,5,6,7-四氢-1H-吲唑-1-基)苯基)氨基)环己基(7-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-4-基)氨基)庚酰基)甘氨酸酯LF7的制备。
制备方法同上述【实施例4】所述,区别在于将原料17a替换为原料17d,得白色固体产物LF7,产率为81%。
1H NMR(400MHz,DMSO-d6)δ11.03(s,1H),8.45(d,J=7.7Hz,1H),8.25(t,J=6.0Hz,1H),8.02(s,1H),7.79(d,J=8.5Hz,1H),7.38(s,1H),7.26(t,J=7.7Hz,1H),6.93–6.88(m,2H),6.75–6.70(m,2H),5.58(t,J=5.5Hz,1H),5.11(dd,J=13.3,5.1Hz,1H),4.71(dt,J=10.2,5.7Hz,1H),4.26–4.07(m,2H),3.78(d,J=5.9Hz,2H),3.46(d,J=8.8Hz,1H),3.09(d,J=6.2Hz,3H),2.98(s,2H),2.94–2.86(m,1H),2.63(d,J=3.8Hz,1H),2.44(s,2H),2.34–2.23(m,1H),2.12(t,J=7.3Hz,2H),2.06–1.98(m,3H),1.94–1.85(m,2H),1.54(dt,J=21.5,6.6Hz,6H),1.34(q,J=11.8,11.1Hz,6H),1.02(s,6H).13C NMR(151MHz,Methanol-d4)δ192.89,176.91,173.90,172.65,171.06,153.45,151.33,145.31,142.88,133.06,131.95,130.75,128.18,117.12,115.79,114.00,112.05,110.80,108.50,74.34,53.70,53.24,47.47,44.49,42.42,37.95,36.77,36.75,32.54,30.37,30.20,30.06,28.37,28.35,28.05,26.92,24.39.HRMS(ESI)m/z calcd for C45H54F3N8O8(M+H)+:891.4017;found,891.4024.
【实施例8】(1R,4R)-4-((2-氨基-5-(6,6-二甲基-4-氧代-3-(三氟甲基)-4,5,6,7-四氢-1H-吲唑-1-基)苯基)氨基)环己基(8-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-4-基)氨基)辛酰基)甘氨酸酯LF8的制备。
制备方法同上述【实施例4】所述,区别在于将原料17a替换为原料17e,得白色固体产物LF8,产率为80%。
1H NMR(400MHz,DMSO-d6)δ11.05(s,1H),8.47(d,J=7.5Hz,1H),8.25(t,J=6.0Hz,1H),8.05(s,1H),7.82(d,J=8.4Hz,1H),7.39(s,1H),7.27(t,J=7.8Hz,1H),6.93(d,J=8.7Hz,2H),6.80–6.68(m,2H),5.58(t,J=5.5Hz,1H),5.13(dd,J=13.3,5.1Hz,1H),4.73(tt,J=10.0,4.1Hz,1H),4.30–4.08(m,2H),3.80(d,J=5.8Hz,2H),3.11(q,J=6.7Hz,2H),2.99(s,2H),2.96–2.85(m,1H),2.64(d,J=17.2Hz,1H),2.46(s,2H),2.31(td,J=13.2,4.5Hz,1H),2.14(t,J=7.3Hz,2H),2.08–1.99(m,3H),1.96–1.87(m,2H),1.61–1.48(m,6H),1.38–1.25(m,8H),1.04(s,6H).13C NMR(101MHz,DMSO-d6)δ190.77,173.45,173.34,171.74,171.28,169.98,169.46,152.43,149.95,144.21,141.13,132.45,131.19,129.70,126.87,119.73,115.80,114.43,112.13,110.36,109.55,107.25,72.58,52.10,51.95,49.14,46.22,43.18,41.31,36.45,35.62,35.47,31.70,29.70,29.52,29.15,29.05,28.96,27.93,27.03,25.64,23.25.HRMS(ESI)m/z calcd for C46H56F3N8O8(M+H)+:905.4173;found,905.4168.
【实施例9】(1R,4R)-4-((2-氨基-5-(6,6-二甲基-4-氧代-3-(三氟甲基)-4,5,6,7-四氢-1H-吲唑-1-基)苯基)氨基)环己基(10-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-4-基)氨基)癸酰基)甘氨酸酯LF9的制备。
制备方法同上述【实施例4】所述,区别在于将原料17a替换为原料17f,得白色固体产物LF9,产率为79%。
1H NMR(400MHz,DMSO-d6)δ11.03(s,1H),8.46(d,J=7.6Hz,1H),8.23(t,J=6.0Hz,1H),8.02(s,1H),7.79(d,J=8.5Hz,1H),7.38(s,1H),7.26(t,J=7.7Hz,1H),6.94–6.87(m,2H),6.75–6.68(m,2H),5.56(t,J=5.6Hz,1H),5.11(dd,J=13.3,5.1Hz,1H),4.71(dt,J=10.1,5.6Hz,1H),4.25–4.07(m,2H),3.77(d,J=5.9Hz,2H),3.45(d,J=8.8Hz,1H),3.08(q,J=6.6Hz,2H),2.98(s,2H),2.94–2.85(m,1H),2.61(d,J=17.4Hz,1H),2.44(s,2H),2.34–2.24(m,1H),2.10(t,J=7.4Hz,2H),2.02(d,J=11.1Hz,3H),1.90(d,J=11.3Hz,2H),1.58–1.44(m,6H),1.35(d,J=11.4Hz,4H),1.29–1.22(m,8H),1.02(s,6H).13CNMR(151MHz,Methanol-d4)δ192.88,176.95,174.91,173.87,172.46,171.05,153.43,151.34,145.30,142.88,133.05,131.96,130.76,130.68,128.14,117.12,113.98,112.01,110.77,108.47,74.31,53.70,53.23,47.50,44.59,42.45,37.96,36.89,36.74,32.54,32.29,30.97,30.88,30.66,30.59,30.50,30.34,30.30,30.26,30.24,28.37,28.24,27.01,24.38.HRMS(ESI)m/z calcd for C48H60F3N8O8(M+H)+:933.4486;found,933.4495.
以上合成的本发明的目标化合物LF1-LF9的化学结构见表1。
【实施例10】Hsp90 PROTAC化合物体外Hsp90降解活性
免疫印迹分析操作步骤:用不同浓度的Hsp90 PROTAC化合物处理SiHa细胞10h,用RIPA缓冲液和SDS-PAGE蛋白上样缓冲液获得全细胞裂解液。用BCA蛋白分析试剂盒(KR0008)分析样品的蛋白浓度,并根据标准蛋白曲线调节电泳用样品的体积。用7.5%或10%的SDS-PAGE凝胶分离蛋白,并转移到厚度为0.45μm的PVDE膜(Millipore,000027346)上。用5%牛血清白蛋白(BSA,KR9048-466-8)或5%脱脂乳在室温下封闭膜2h,一级抗体在4℃孵育12h以上,二级抗体在摇床上室温孵育1h以上。采用超灵敏增强化学发光(ECL、meilunbio、MAO186-2)试剂检测蛋白质。HSP90抗体(4877S)购自Cell SignalingTechnology(CST),GAPDH(60004-1-Ig)抗体购自Proteintech。
结果如图1所示,表明大多数Hsp90 PROTAC化合物均能诱导Hsp90降解,例如LF1以及LF6-LF8。其中,化合物LF8在10h内能显著诱导Hsp90降解,在浓度仅为0.05μM,能够显著降低Hsp90的蛋白水平,然而当浓度升高到1μM时,Hsp90的水平开始回升,出现了明显的“hook”效应。
【实施例11】Hsp90 PROTAC化合物体外抗宫颈癌活性
CCK-8用于化合物抗增殖活性测试原理:CCK-8(Cell Counting Kit-8)是一种基于WST-8的广泛应用于细胞增殖和细胞毒性的检测试剂。WST-8(化学名:2-(2-甲氧基-4-硝基苯基)-3-(4-硝基苯基)-5-(2,4-二磺酸苯)-2H-四唑单钠盐),是一种类似于MTT(化学名:3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐)的化合物,其在电子载体1-甲氧基-5-甲基吩嗪鎓硫酸二甲酯的作用下,被线粒体中的脱氢酶还原为具有高度水溶性的橙黄色甲瓒产物(Formazan)。细胞增殖数量越多、速度越快,颜色越深;细胞毒性越大,则颜色越浅,对于同样的细胞,颜色的深浅与活细胞的数量成正比,因此可利用这一特性直接进行细胞增殖和毒性分析。
操作步骤:
1)制备细胞悬液:宫颈癌细胞(SiHa、HaLa、C33A或CaSKi)生长密度达到80%-90%时,用0.25% Trypsin-EDTA消化细胞,加入新鲜的DMEM培养基,混匀细胞悬液,使用细胞计数板计数,将细胞稀释为5×104个/ml的单细胞悬液。
2)铺板:将100μL单细胞悬液接种到微孔板(组织培养级,96孔,平底)中。
3)预培养:在37℃、5% CO2培养箱中孵育细胞24小时左右。
4)加药:吸出原培养基,向培养板各孔中依次加入200μL含不同浓度(100μM-0.01μM)待测药物的培养基(每个浓度设置3个复孔,同时设置空白组和顺铂对照组),放入培养箱孵育72h。
6)收板:按照每100μL培养基加10μL CCK-8试剂的比例配制CCK-8工作液,每孔中加入100μL CCK-8工作液,将培养板置于培养箱中孵育1-2h。
7)测板:用酶标仪测定450nm处的吸光度(OD)。
表2.Hsp90 PROTAC化合物(LF1-LF9)的抗宫颈癌细胞增殖活性
上述实验结果表明:合成的大多数Hsp90 PROTAC化合物在上述四种宫颈癌细胞中都具有很好抗宫颈癌活性,且优于阳性对照药物顺铂(IC50=200-1290nM),其中部分化合物的抑制活性达到了纳摩尔水平,例如化合物LF2(IC50=40-170nM)、LF4(IC50=60-120nM)、LF7(IC50=90-250nM)、LF8(IC50=50-80nM)、LF9(IC50=60-150nM)。
上述实施例仅为本发明较佳的实施方式,本发明的实施方式并不受上述实施例的限制,其他的任何未背离本发明的精神实质与原理下所作的改变、修饰、替代、组合、简化,均应为等效的置换方式,都包含在本发明的保护范围之内。
Claims (10)
1.一种Hsp90 PROTAC化合物,其特征在于,结构如通式(I)所示:
其中,
Linker选自
E3 ligand选自
2.根据权利要求1所述的Hsp90 PROTAC化合物,其特征在于,选自下列化合物:
(1R,4S)-4-((2-氨基甲酰基-5-(6,6-二甲基-4-氧代-3-(三氟甲基)-4,5,6,7-四氢-1H-吲唑-1-基)苯基)氨基)环己基(6-(((S)-1-((2S,4R)-4-羟基-2-((((S)-1-(4-(4-甲基噻唑-5-基)苯)乙基)氨基甲酰基)吡咯烷-1-基)-3,3-二甲基-1-氧代丁-2-基)氨基)己酰基)甘氨酸酯;
(1R,4R)-4-((2-氨基甲酰基-5-(6,6-二甲基-4-氧代-3-(三氟甲基)-4,5,6,7-四氢-1H-吲唑-1-基)苯基)氨基)环己基(8-(((S)-1-((2S,4R)-4-羟基-2-((((S)-1-(4-(4-甲基噻唑-5-基)苯)乙基)氨基甲酰基)吡咯烷-1-基)-3,3-二甲基-1-氧代丁-2-基)氨基)辛酰基)甘氨酸酯;
(1R,4R)-4-((2-氨基甲酰基-5-(6,6-二甲基-4-氧代-3-(三氟甲基)-4,5,6,7-四氢-1H-吲唑-1-基)苯基)氨基)环己基(10-(((S)-1-((2S,4r)-4-羟基-2-((((S)-1-(4-(4-甲基噻唑-5-基)苯)乙基)氨基甲酰基)吡咯烷-1-基)-3,3-二甲基-1-氧代丁-2-基)氨基)癸酰基)甘氨酸酯;
(1R,4R)-4-((2-氨基-5-(6,6-二甲基-4-氧代-3-(三氟甲基)-4,5,6,7-四氢-1H-吲唑-1-基)苯基)氨基)环己基(4-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-4-基)氨基)丁酰基)甘氨酸酯;
(1R,4R)-4-((2-氨基-5-(6,6-二甲基-4-氧代-3-(三氟甲基)-4,5,6,7-四氢-1H-吲唑-1-基)苯基)氨基)环己基(5-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-4-基)氨基)戊酰基)甘氨酸酯;
(1R,4R)-4-((2-氨基-5-(6,6-二甲基-4-氧代-3-(三氟甲基)-4,5,6,7-四氢-1H-吲唑-1-基)苯基)氨基)环己基(6-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-4-基)氨基)己酰基)甘氨酸酯;
(1R,4R)-4-((2-氨基-5-(6,6-二甲基-4-氧代-3-(三氟甲基)-4,5,6,7-四氢-1H-吲唑-1-基)苯基)氨基)环己基(7-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-4-基)氨基)庚酰基)甘氨酸酯;
(1R,4R)-4-((2-氨基-5-(6,6-二甲基-4-氧代-3-(三氟甲基)-4,5,6,7-四氢-1H-吲唑-1-基)苯基)氨基)环己基(8-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-4-基)氨基)辛酰基)甘氨酸酯;
(1R,4R)-4-((2-氨基-5-(6,6-二甲基-4-氧代-3-(三氟甲基)-4,5,6,7-四氢-1H-吲唑-1-基)苯基)氨基)环己基(10-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-4-基)氨基)癸酰基)甘氨酸酯。
3.根据权利要求1所述的Hsp90 PROTAC化合物,其特征在于,选自下列化合物:
(1R,4R)-4-((2-氨基甲酰基-5-(6,6-二甲基-4-氧代-3-(三氟甲基)-4,5,6,7-四氢-1H-吲唑-1-基)苯基)氨基)环己基(8-(((S)-1-((2S,4R)-4-羟基-2-((((S)-1-(4-(4-甲基噻唑-5-基)苯)乙基)氨基甲酰基)吡咯烷-1-基)-3,3-二甲基-1-氧代丁-2-基)氨基)辛酰基)甘氨酸酯;
(1R,4R)-4-((2-氨基-5-(6,6-二甲基-4-氧代-3-(三氟甲基)-4,5,6,7-四氢-1H-吲唑-1-基)苯基)氨基)环己基(4-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-4-基)氨基)丁酰基)甘氨酸酯;
(1R,4R)-4-((2-氨基-5-(6,6-二甲基-4-氧代-3-(三氟甲基)-4,5,6,7-四氢-1H-吲唑-1-基)苯基)氨基)环己基(7-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-4-基)氨基)庚酰基)甘氨酸酯;
(1R,4R)-4-((2-氨基-5-(6,6-二甲基-4-氧代-3-(三氟甲基)-4,5,6,7-四氢-1H-吲唑-1-基)苯基)氨基)环己基(8-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-4-基)氨基)辛酰基)甘氨酸酯;
(1R,4R)-4-((2-氨基-5-(6,6-二甲基-4-氧代-3-(三氟甲基)-4,5,6,7-四氢-1H-吲唑-1-基)苯基)氨基)环己基(10-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-4-基)氨基)癸酰基)甘氨酸酯。
4.一种权利要求1-3任一项所述的Hsp90 PROTAC化合物药理或生理上可接受的盐。
5.权利要求1-3任一项所述的Hsp90 PROTAC化合物或其药理或生理上可接受的盐在制备Hsp90降解剂或抗宫颈癌的药物中的应用。
6.一种Hsp90降解剂或抗宫颈癌的药物,其特征在于:包含权利要求1-3任一项所述的Hsp90 PROTAC化合物或其药理或生理上可接受的盐中的一种或多种。
7.根据权利要求6所述的Hsp90降解剂或抗宫颈癌的药物,其特征在于:还包含药学上可接受的载体或赋形剂。
8.权利要求1-3任一项所述的芳基甲酰胺类化合物的制备方法,其特征在于,包括如下步骤:含羧酸连接子的VHL或来那度胺衍生物与SNX-5422在缩合剂和碱的作用下进行缩合反应,得到所述的Hsp90 PROTAC化合物;
所述的含羧酸连接子的VHL或来那度胺衍生物为 其中,n=1、2、3、4、5、7。
9.根据权利要求8所述的芳基甲酰胺类化合物的制备方法,其特征在于:所述的缩合剂为HATU,所述的碱为EIPEA。
10.根据权利要求9所述的芳基甲酰胺类化合物的制备方法,其特征在于:SNX-5422、含羧酸连接子的VHL或来那度胺衍生物、HATU和DIPEA投料的摩尔比为1:1.0~1.2:1.0~1.2:2.5~3.5。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202311816125.7A CN117820416A (zh) | 2023-12-27 | 2023-12-27 | 一种Hsp90 PROTAC化合物及其制备方法与在制备抗宫颈癌药物中的应用 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202311816125.7A CN117820416A (zh) | 2023-12-27 | 2023-12-27 | 一种Hsp90 PROTAC化合物及其制备方法与在制备抗宫颈癌药物中的应用 |
Publications (1)
Publication Number | Publication Date |
---|---|
CN117820416A true CN117820416A (zh) | 2024-04-05 |
Family
ID=90518328
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202311816125.7A Pending CN117820416A (zh) | 2023-12-27 | 2023-12-27 | 一种Hsp90 PROTAC化合物及其制备方法与在制备抗宫颈癌药物中的应用 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN117820416A (zh) |
-
2023
- 2023-12-27 CN CN202311816125.7A patent/CN117820416A/zh active Pending
Similar Documents
Publication | Publication Date | Title |
---|---|---|
KR20190122678A (ko) | 신규한 이소인돌린 유도체, 이의 약학 조성물 및 용도 | |
US8946249B2 (en) | Compound, certain novel forms thereof, pharmaceutical compositions thereof and methods for preparation and use | |
CN112300153B (zh) | 一种杂环化合物、药物组合物和用途 | |
CN104230827A (zh) | 1-(芳基甲基)喹唑啉-2,4(1h,3h)-二酮作为parp抑制剂及其应用 | |
CN115353508B (zh) | 5-吡啶-1h-吲唑类化合物、药物组合物和应用 | |
CN114920704A (zh) | 一种苯基哌嗪喹唑啉类化合物或其药学上可接受的盐、制法与用途 | |
WO2005082001A2 (en) | Advanced isothiazole based protein kinase inhibitors | |
CA2619459A1 (en) | Novel 4-amino-thieno[3,2-c]pyridine-7-carboxylic acid amides | |
CN112457365B (zh) | 一类靶向蛋白质水解通路的功能分子及其制备和应用 | |
CN117820416A (zh) | 一种Hsp90 PROTAC化合物及其制备方法与在制备抗宫颈癌药物中的应用 | |
CN108456214B (zh) | 含噁唑或咪唑结构的喹唑啉类化合物及其应用 | |
CN114605407B (zh) | 一种吲哚喹啉酮类化合物及其合成方法和应用 | |
CN115197167A (zh) | 1,2,4-噻二唑烷-3,5-二酮化合物及其制备方法和应用 | |
CN110981865B (zh) | 一种用于治疗脑胶质瘤的药物及其制备方法 | |
CN106008394A (zh) | 巯基苯并噻唑酰胺类化合物及其制备与作为药物的用途 | |
CN107501283B (zh) | 取代芳甲杂基取代苯胺基缩乙二醇醚环代喹唑啉的制备及肿瘤治疗药物应用 | |
CN114907387B (zh) | 嘧啶并吡咯类kras抑制剂及其制备方法与应用 | |
CN116444499B (zh) | 一种含4-喹唑啉酮的苯丙氨酸类衍生物及其制备方法与应用 | |
CN110590807B (zh) | 一种噻吩并[3,2-d]嘧啶类衍生物及其应用 | |
CN113461684B (zh) | 一种PPAR-γ配体—4,5-二氮芴-罗丹宁轭合物及其制备方法和抗肿瘤应用 | |
CN114380728B (zh) | 新型二芳基-β-内酰胺类有机硒化合物及其制备方法和在制药中的用途 | |
CN111217821B (zh) | 系列二噁烷并喹唑啉衍生物的制备方法 | |
CN114716435A (zh) | Kras g12c突变抑制剂 | |
CN116554182A (zh) | 一种卟啉-丁香酚衍生物、制备方法及其用途 | |
CN118440052A (zh) | 一种化合物及其在制备tyk2激酶抑制剂中的用途 |
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 |