CN113651835A - 一种bethdac双靶向protac的制备方法和应用 - Google Patents
一种bethdac双靶向protac的制备方法和应用 Download PDFInfo
- Publication number
- CN113651835A CN113651835A CN202110627242.3A CN202110627242A CN113651835A CN 113651835 A CN113651835 A CN 113651835A CN 202110627242 A CN202110627242 A CN 202110627242A CN 113651835 A CN113651835 A CN 113651835A
- Authority
- CN
- China
- Prior art keywords
- compound
- cancer
- protac
- dioxopiperidin
- diazepin
- 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
- 238000011865 proteolysis targeting chimera technique Methods 0.000 title claims abstract description 22
- 229940124823 proteolysis targeting chimeric molecule Drugs 0.000 title claims abstract description 22
- 108010026668 snake venom protein C activator Proteins 0.000 title claims abstract description 22
- 238000002360 preparation method Methods 0.000 title claims abstract description 9
- 150000001875 compounds Chemical class 0.000 claims abstract description 69
- 102000003964 Histone deacetylase Human genes 0.000 claims abstract description 24
- 108090000353 Histone deacetylase Proteins 0.000 claims abstract description 24
- 108091005625 BRD4 Proteins 0.000 claims abstract description 22
- 102100029895 Bromodomain-containing protein 4 Human genes 0.000 claims abstract description 22
- 206010028980 Neoplasm Diseases 0.000 claims abstract description 11
- 239000003276 histone deacetylase inhibitor Substances 0.000 claims abstract description 4
- -1 piperazinonyl Chemical group 0.000 claims description 29
- WWZKQHOCKIZLMA-UHFFFAOYSA-N octanoic acid Chemical compound CCCCCCCC(O)=O WWZKQHOCKIZLMA-UHFFFAOYSA-N 0.000 claims description 13
- 238000000034 method Methods 0.000 claims description 9
- 150000003839 salts Chemical class 0.000 claims description 9
- 239000003112 inhibitor Substances 0.000 claims description 7
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 6
- 239000002253 acid Substances 0.000 claims description 6
- 125000000623 heterocyclic group Chemical group 0.000 claims description 6
- 125000005647 linker group Chemical group 0.000 claims description 6
- 206010006187 Breast cancer Diseases 0.000 claims description 5
- 208000026310 Breast neoplasm Diseases 0.000 claims description 5
- 230000008569 process Effects 0.000 claims description 5
- 210000004881 tumor cell Anatomy 0.000 claims description 5
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical group CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 4
- 206010058467 Lung neoplasm malignant Diseases 0.000 claims description 4
- LCTONWCANYUPML-UHFFFAOYSA-N Pyruvic acid Chemical compound CC(=O)C(O)=O LCTONWCANYUPML-UHFFFAOYSA-N 0.000 claims description 4
- FEWJPZIEWOKRBE-UHFFFAOYSA-N Tartaric Acid Chemical compound [H+].[H+].[O-]C(=O)C(O)C(O)C([O-])=O FEWJPZIEWOKRBE-UHFFFAOYSA-N 0.000 claims description 4
- OBETXYAYXDNJHR-UHFFFAOYSA-N alpha-ethylcaproic acid Natural products CCCCC(CC)C(O)=O OBETXYAYXDNJHR-UHFFFAOYSA-N 0.000 claims description 4
- 125000004432 carbon atom Chemical group C* 0.000 claims description 4
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 4
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 claims description 4
- 125000002541 furyl group Chemical group 0.000 claims description 4
- 125000001786 isothiazolyl group Chemical group 0.000 claims description 4
- 125000000842 isoxazolyl group Chemical group 0.000 claims description 4
- 201000007270 liver cancer Diseases 0.000 claims description 4
- 208000014018 liver neoplasm Diseases 0.000 claims description 4
- 201000005202 lung cancer Diseases 0.000 claims description 4
- 208000020816 lung neoplasm Diseases 0.000 claims description 4
- 125000000449 nitro group Chemical group [O-][N+](*)=O 0.000 claims description 4
- 125000002971 oxazolyl group Chemical group 0.000 claims description 4
- 125000003373 pyrazinyl group Chemical group 0.000 claims description 4
- 125000003226 pyrazolyl group Chemical group 0.000 claims description 4
- 125000002098 pyridazinyl group Chemical group 0.000 claims description 4
- 125000004076 pyridyl group Chemical group 0.000 claims description 4
- 125000000714 pyrimidinyl group Chemical group 0.000 claims description 4
- 125000000168 pyrrolyl group Chemical group 0.000 claims description 4
- 229920006395 saturated elastomer Polymers 0.000 claims description 4
- 125000000335 thiazolyl group Chemical group 0.000 claims description 4
- 125000001544 thienyl group Chemical group 0.000 claims description 4
- 206010025323 Lymphomas Diseases 0.000 claims description 3
- 206010061902 Pancreatic neoplasm Diseases 0.000 claims description 3
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 claims description 3
- 201000010099 disease Diseases 0.000 claims description 3
- QBYIENPQHBMVBV-HFEGYEGKSA-N (2R)-2-hydroxy-2-phenylacetic acid Chemical compound O[C@@H](C(O)=O)c1ccccc1.O[C@@H](C(O)=O)c1ccccc1 QBYIENPQHBMVBV-HFEGYEGKSA-N 0.000 claims description 2
- BMYNFMYTOJXKLE-UHFFFAOYSA-N 3-azaniumyl-2-hydroxypropanoate Chemical compound NCC(O)C(O)=O BMYNFMYTOJXKLE-UHFFFAOYSA-N 0.000 claims description 2
- PXRKCOCTEMYUEG-UHFFFAOYSA-N 5-aminoisoindole-1,3-dione Chemical compound NC1=CC=C2C(=O)NC(=O)C2=C1 PXRKCOCTEMYUEG-UHFFFAOYSA-N 0.000 claims description 2
- 206010005003 Bladder cancer Diseases 0.000 claims description 2
- 239000004215 Carbon black (E152) Substances 0.000 claims description 2
- 208000000461 Esophageal Neoplasms Diseases 0.000 claims description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical group CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 2
- 206010017993 Gastrointestinal neoplasms Diseases 0.000 claims description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 2
- 208000008839 Kidney Neoplasms Diseases 0.000 claims description 2
- AFVFQIVMOAPDHO-UHFFFAOYSA-N Methanesulfonic acid Chemical compound CS(O)(=O)=O AFVFQIVMOAPDHO-UHFFFAOYSA-N 0.000 claims description 2
- 201000003793 Myelodysplastic syndrome Diseases 0.000 claims description 2
- 206010030155 Oesophageal carcinoma Diseases 0.000 claims description 2
- 206010033128 Ovarian cancer Diseases 0.000 claims description 2
- 206010061535 Ovarian neoplasm Diseases 0.000 claims description 2
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical group OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 claims description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 claims description 2
- OFOBLEOULBTSOW-UHFFFAOYSA-N Propanedioic acid Natural products OC(=O)CC(O)=O OFOBLEOULBTSOW-UHFFFAOYSA-N 0.000 claims description 2
- 206010060862 Prostate cancer Diseases 0.000 claims description 2
- 208000000236 Prostatic Neoplasms Diseases 0.000 claims description 2
- IWYDHOAUDWTVEP-UHFFFAOYSA-N R-2-phenyl-2-hydroxyacetic acid Natural products OC(=O)C(O)C1=CC=CC=C1 IWYDHOAUDWTVEP-UHFFFAOYSA-N 0.000 claims description 2
- 206010038389 Renal cancer Diseases 0.000 claims description 2
- 229910006069 SO3H Inorganic materials 0.000 claims description 2
- 208000000453 Skin Neoplasms Diseases 0.000 claims description 2
- KDYFGRWQOYBRFD-UHFFFAOYSA-N Succinic acid Natural products OC(=O)CCC(O)=O KDYFGRWQOYBRFD-UHFFFAOYSA-N 0.000 claims description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 claims description 2
- 208000024770 Thyroid neoplasm Diseases 0.000 claims description 2
- 208000007097 Urinary Bladder Neoplasms Diseases 0.000 claims description 2
- CIUQDSCDWFSTQR-UHFFFAOYSA-N [C]1=CC=CC=C1 Chemical compound [C]1=CC=CC=C1 CIUQDSCDWFSTQR-UHFFFAOYSA-N 0.000 claims description 2
- 125000000738 acetamido group Chemical group [H]C([H])([H])C(=O)N([H])[*] 0.000 claims description 2
- 150000001336 alkenes Chemical group 0.000 claims description 2
- 150000001408 amides Chemical class 0.000 claims description 2
- 150000001412 amines Chemical class 0.000 claims description 2
- SRSXLGNVWSONIS-UHFFFAOYSA-N benzenesulfonic acid Chemical compound OS(=O)(=O)C1=CC=CC=C1 SRSXLGNVWSONIS-UHFFFAOYSA-N 0.000 claims description 2
- KDYFGRWQOYBRFD-NUQCWPJISA-N butanedioic acid Chemical compound O[14C](=O)CC[14C](O)=O KDYFGRWQOYBRFD-NUQCWPJISA-N 0.000 claims description 2
- 210000003169 central nervous system Anatomy 0.000 claims description 2
- 208000024558 digestive system cancer Diseases 0.000 claims description 2
- 201000004101 esophageal cancer Diseases 0.000 claims description 2
- 125000004185 ester group Chemical group 0.000 claims description 2
- 125000001301 ethoxy group Chemical group [H]C([H])([H])C([H])([H])O* 0.000 claims description 2
- 125000002485 formyl group Chemical group [H]C(*)=O 0.000 claims description 2
- 201000010231 gastrointestinal system cancer Diseases 0.000 claims description 2
- 229910052736 halogen Inorganic materials 0.000 claims description 2
- 125000005843 halogen group Chemical group 0.000 claims description 2
- 229930195733 hydrocarbon Natural products 0.000 claims description 2
- 150000002576 ketones Chemical group 0.000 claims description 2
- 201000010982 kidney cancer Diseases 0.000 claims description 2
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 claims description 2
- 239000011976 maleic acid Substances 0.000 claims description 2
- VZCYOOQTPOCHFL-UPHRSURJSA-N maleic acid Chemical compound OC(=O)\C=C/C(O)=O VZCYOOQTPOCHFL-UPHRSURJSA-N 0.000 claims description 2
- 208000015486 malignant pancreatic neoplasm Diseases 0.000 claims description 2
- 229960002510 mandelic acid Drugs 0.000 claims description 2
- PSZYNBSKGUBXEH-UHFFFAOYSA-N naphthalene-1-sulfonic acid Chemical compound C1=CC=C2C(S(=O)(=O)O)=CC=CC2=C1 PSZYNBSKGUBXEH-UHFFFAOYSA-N 0.000 claims description 2
- 208000002154 non-small cell lung carcinoma Diseases 0.000 claims description 2
- JRZJOMJEPLMPRA-UHFFFAOYSA-N olefin Chemical group CCCCCCCC=C JRZJOMJEPLMPRA-UHFFFAOYSA-N 0.000 claims description 2
- 239000005416 organic matter Substances 0.000 claims description 2
- 201000008968 osteosarcoma Diseases 0.000 claims description 2
- 201000002528 pancreatic cancer Diseases 0.000 claims description 2
- 208000008443 pancreatic carcinoma Diseases 0.000 claims description 2
- 210000001428 peripheral nervous system Anatomy 0.000 claims description 2
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 claims description 2
- WLJVXDMOQOGPHL-UHFFFAOYSA-N phenylacetic acid Chemical compound OC(=O)CC1=CC=CC=C1 WLJVXDMOQOGPHL-UHFFFAOYSA-N 0.000 claims description 2
- 125000004193 piperazinyl group Chemical group 0.000 claims description 2
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 claims description 2
- 201000000849 skin cancer Diseases 0.000 claims description 2
- 201000002510 thyroid cancer Diseases 0.000 claims description 2
- JOXIMZWYDAKGHI-UHFFFAOYSA-N toluene-4-sulfonic acid Chemical compound CC1=CC=C(S(O)(=O)=O)C=C1 JOXIMZWYDAKGHI-UHFFFAOYSA-N 0.000 claims description 2
- VZCYOOQTPOCHFL-UHFFFAOYSA-N trans-butenedioic acid Natural products OC(=O)C=CC(O)=O VZCYOOQTPOCHFL-UHFFFAOYSA-N 0.000 claims description 2
- 208000029729 tumor suppressor gene on chromosome 11 Diseases 0.000 claims description 2
- 201000005112 urinary bladder cancer Diseases 0.000 claims description 2
- 150000001735 carboxylic acids Chemical group 0.000 claims 1
- 150000002430 hydrocarbons Chemical group 0.000 claims 1
- 238000011580 nude mouse model Methods 0.000 claims 1
- 230000004614 tumor growth Effects 0.000 claims 1
- 230000005909 tumor killing Effects 0.000 claims 1
- 230000000694 effects Effects 0.000 abstract description 5
- 230000005764 inhibitory process Effects 0.000 abstract description 5
- 238000011160 research Methods 0.000 abstract description 5
- 238000011161 development Methods 0.000 abstract description 4
- 230000005918 in vitro anti-tumor Effects 0.000 abstract description 4
- 102000006275 Ubiquitin-Protein Ligases Human genes 0.000 abstract description 3
- 108010083111 Ubiquitin-Protein Ligases Proteins 0.000 abstract description 3
- 238000002474 experimental method Methods 0.000 abstract description 3
- 230000007246 mechanism Effects 0.000 abstract description 2
- 230000001404 mediated effect Effects 0.000 abstract description 2
- 230000035755 proliferation Effects 0.000 abstract description 2
- 229940125763 bromodomain inhibitor Drugs 0.000 abstract 1
- 229940121372 histone deacetylase inhibitor Drugs 0.000 abstract 1
- 230000001575 pathological effect Effects 0.000 abstract 1
- 230000000144 pharmacologic effect Effects 0.000 abstract 1
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 24
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 21
- 108090000623 proteins and genes Proteins 0.000 description 20
- 102000004169 proteins and genes Human genes 0.000 description 19
- 238000006243 chemical reaction Methods 0.000 description 13
- 230000002401 inhibitory effect Effects 0.000 description 13
- 230000015572 biosynthetic process Effects 0.000 description 11
- 238000003786 synthesis reaction Methods 0.000 description 11
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 10
- 210000004027 cell Anatomy 0.000 description 10
- JGFZNNIVVJXRND-UHFFFAOYSA-N N,N-Diisopropylethylamine (DIPEA) Chemical compound CCN(C(C)C)C(C)C JGFZNNIVVJXRND-UHFFFAOYSA-N 0.000 description 9
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 description 9
- 230000002829 reductive effect Effects 0.000 description 8
- 239000002904 solvent Substances 0.000 description 8
- 230000000259 anti-tumor effect Effects 0.000 description 7
- 235000018102 proteins Nutrition 0.000 description 7
- 102100039996 Histone deacetylase 1 Human genes 0.000 description 6
- 101001035024 Homo sapiens Histone deacetylase 1 Proteins 0.000 description 6
- 239000012071 phase Substances 0.000 description 6
- 238000012360 testing method Methods 0.000 description 5
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 4
- GOTYRUGSSMKFNF-UHFFFAOYSA-N lenalidomide Chemical compound C1C=2C(N)=CC=CC=2C(=O)N1C1CCC(=O)NC1=O GOTYRUGSSMKFNF-UHFFFAOYSA-N 0.000 description 4
- 239000000741 silica gel Substances 0.000 description 4
- 229910002027 silica gel Inorganic materials 0.000 description 4
- 239000007787 solid Substances 0.000 description 4
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 3
- 239000007821 HATU Substances 0.000 description 3
- 230000001588 bifunctional effect Effects 0.000 description 3
- 230000000903 blocking effect Effects 0.000 description 3
- 238000006482 condensation reaction Methods 0.000 description 3
- 238000013461 design Methods 0.000 description 3
- 230000018109 developmental process Effects 0.000 description 3
- 230000003828 downregulation Effects 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 239000005457 ice water Substances 0.000 description 3
- 239000003446 ligand Substances 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 239000012074 organic phase Substances 0.000 description 3
- 125000003854 p-chlorophenyl group Chemical group [H]C1=C([H])C(*)=C([H])C([H])=C1Cl 0.000 description 3
- 238000005160 1H NMR spectroscopy Methods 0.000 description 2
- IFSMYFLQPDFUOI-UHFFFAOYSA-N 2-[2-[2-[2-[(2-methylpropan-2-yl)oxycarbonylamino]ethoxy]ethoxy]ethoxy]acetic acid Chemical compound CC(C)(C)OC(=O)NCCOCCOCCOCC(O)=O IFSMYFLQPDFUOI-UHFFFAOYSA-N 0.000 description 2
- GNFTZDOKVXKIBK-UHFFFAOYSA-N 3-(2-methoxyethoxy)benzohydrazide Chemical compound COCCOC1=CC=CC(C(=O)NN)=C1 GNFTZDOKVXKIBK-UHFFFAOYSA-N 0.000 description 2
- FGUUSXIOTUKUDN-IBGZPJMESA-N C1(=CC=CC=C1)N1C2=C(NC([C@H](C1)NC=1OC(=NN=1)C1=CC=CC=C1)=O)C=CC=C2 Chemical compound C1(=CC=CC=C1)N1C2=C(NC([C@H](C1)NC=1OC(=NN=1)C1=CC=CC=C1)=O)C=CC=C2 FGUUSXIOTUKUDN-IBGZPJMESA-N 0.000 description 2
- WMFOQBRAJBCJND-UHFFFAOYSA-M Lithium hydroxide Chemical compound [Li+].[OH-] WMFOQBRAJBCJND-UHFFFAOYSA-M 0.000 description 2
- PXIPVTKHYLBLMZ-UHFFFAOYSA-N Sodium azide Chemical compound [Na+].[N-]=[N+]=[N-] PXIPVTKHYLBLMZ-UHFFFAOYSA-N 0.000 description 2
- DKGAVHZHDRPRBM-UHFFFAOYSA-N Tert-Butanol Chemical compound CC(C)(C)O DKGAVHZHDRPRBM-UHFFFAOYSA-N 0.000 description 2
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 description 2
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 description 2
- 201000011510 cancer Diseases 0.000 description 2
- 230000015556 catabolic process Effects 0.000 description 2
- LJOSBOOJFIRCSO-AWEZNQCLSA-N cs-m2721 Chemical compound N([C@@H](CC(O)=O)C1=NN=C(N1C=1SC(C)=C(C)C=11)C)=C1C1=CC=C(Cl)C=C1 LJOSBOOJFIRCSO-AWEZNQCLSA-N 0.000 description 2
- 238000006731 degradation reaction Methods 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 230000009977 dual effect Effects 0.000 description 2
- 230000001973 epigenetic effect Effects 0.000 description 2
- 238000002875 fluorescence polarization Methods 0.000 description 2
- 230000014509 gene expression Effects 0.000 description 2
- 238000007429 general method Methods 0.000 description 2
- 238000000338 in vitro Methods 0.000 description 2
- 229960004942 lenalidomide Drugs 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- NZFHJBSDSXDUAO-UHFFFAOYSA-N o-tritylhydroxylamine Chemical compound C=1C=CC=CC=1C(C=1C=CC=CC=1)(ON)C1=CC=CC=C1 NZFHJBSDSXDUAO-UHFFFAOYSA-N 0.000 description 2
- 239000013641 positive control Substances 0.000 description 2
- BWHMMNNQKKPAPP-UHFFFAOYSA-L potassium carbonate Chemical compound [K+].[K+].[O-]C([O-])=O BWHMMNNQKKPAPP-UHFFFAOYSA-L 0.000 description 2
- 125000006239 protecting group Chemical group 0.000 description 2
- 230000000717 retained effect Effects 0.000 description 2
- 238000010898 silica gel chromatography Methods 0.000 description 2
- 150000003384 small molecules Chemical class 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 239000007858 starting material Substances 0.000 description 2
- 230000002195 synergetic effect Effects 0.000 description 2
- 125000000999 tert-butyl group Chemical group [H]C([H])([H])C(*)(C([H])([H])[H])C([H])([H])[H] 0.000 description 2
- LWIHDJKSTIGBAC-UHFFFAOYSA-K tripotassium phosphate Chemical compound [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 description 2
- 238000010626 work up procedure Methods 0.000 description 2
- 238000001644 13C nuclear magnetic resonance spectroscopy Methods 0.000 description 1
- NEAQRZUHTPSBBM-UHFFFAOYSA-N 2-hydroxy-3,3-dimethyl-7-nitro-4h-isoquinolin-1-one Chemical compound C1=C([N+]([O-])=O)C=C2C(=O)N(O)C(C)(C)CC2=C1 NEAQRZUHTPSBBM-UHFFFAOYSA-N 0.000 description 1
- 206010000830 Acute leukaemia Diseases 0.000 description 1
- 102100021569 Apoptosis regulator Bcl-2 Human genes 0.000 description 1
- 108091012583 BCL2 Proteins 0.000 description 1
- 201000009030 Carcinoma Diseases 0.000 description 1
- 102400000888 Cholecystokinin-8 Human genes 0.000 description 1
- 101800005151 Cholecystokinin-8 Proteins 0.000 description 1
- 208000017667 Chronic Disease Diseases 0.000 description 1
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000006947 Histones Human genes 0.000 description 1
- 108010033040 Histones Proteins 0.000 description 1
- 101001030211 Homo sapiens Myc proto-oncogene protein Proteins 0.000 description 1
- 206010027476 Metastases Diseases 0.000 description 1
- 101100381525 Mus musculus Bcl6 gene Proteins 0.000 description 1
- 101710135898 Myc proto-oncogene protein Proteins 0.000 description 1
- 102100038895 Myc proto-oncogene protein Human genes 0.000 description 1
- 108700020796 Oncogene Proteins 0.000 description 1
- 102000043276 Oncogene Human genes 0.000 description 1
- 108010019160 Pancreatin Proteins 0.000 description 1
- 108091000080 Phosphotransferase Proteins 0.000 description 1
- 102000011990 Sirtuin Human genes 0.000 description 1
- 108050002485 Sirtuin Proteins 0.000 description 1
- 101710150448 Transcriptional regulator Myc Proteins 0.000 description 1
- 102000044209 Tumor Suppressor Genes Human genes 0.000 description 1
- 108700025716 Tumor Suppressor Genes Proteins 0.000 description 1
- 125000002777 acetyl group Chemical group [H]C([H])([H])C(*)=O 0.000 description 1
- 150000007513 acids Chemical class 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000003698 anagen phase Effects 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000006907 apoptotic process Effects 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- NCNRHFGMJRPRSK-MDZDMXLPSA-N belinostat Chemical compound ONC(=O)\C=C\C1=CC=CC(S(=O)(=O)NC=2C=CC=CC=2)=C1 NCNRHFGMJRPRSK-MDZDMXLPSA-N 0.000 description 1
- 239000012964 benzotriazole Substances 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 150000001732 carboxylic acid derivatives Chemical group 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 230000009918 complex formation Effects 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- DEZRYPDIMOWBDS-UHFFFAOYSA-N dcm dichloromethane Chemical compound ClCCl.ClCCl DEZRYPDIMOWBDS-UHFFFAOYSA-N 0.000 description 1
- 230000006196 deacetylation Effects 0.000 description 1
- 238000003381 deacetylation reaction Methods 0.000 description 1
- 230000000593 degrading effect Effects 0.000 description 1
- 230000001627 detrimental effect Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 229940088598 enzyme Drugs 0.000 description 1
- 150000002148 esters Chemical group 0.000 description 1
- 238000001704 evaporation Methods 0.000 description 1
- 230000008020 evaporation Effects 0.000 description 1
- 230000005284 excitation Effects 0.000 description 1
- 230000004927 fusion Effects 0.000 description 1
- 201000005787 hematologic cancer Diseases 0.000 description 1
- 230000002489 hematologic effect Effects 0.000 description 1
- 125000001183 hydrocarbyl group Chemical group 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000008595 infiltration Effects 0.000 description 1
- 238000001764 infiltration Methods 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 230000002147 killing effect Effects 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 235000018977 lysine Nutrition 0.000 description 1
- 150000002669 lysines Chemical class 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 230000009401 metastasis Effects 0.000 description 1
- HDZGCSFEDULWCS-UHFFFAOYSA-N monomethylhydrazine Chemical compound CNN HDZGCSFEDULWCS-UHFFFAOYSA-N 0.000 description 1
- 230000009437 off-target effect Effects 0.000 description 1
- 230000002018 overexpression Effects 0.000 description 1
- 229940055695 pancreatin Drugs 0.000 description 1
- FPOHNWQLNRZRFC-ZHACJKMWSA-N panobinostat Chemical compound CC=1NC2=CC=CC=C2C=1CCNCC1=CC=C(\C=C\C(=O)NO)C=C1 FPOHNWQLNRZRFC-ZHACJKMWSA-N 0.000 description 1
- 230000036961 partial effect Effects 0.000 description 1
- 102000020233 phosphotransferase Human genes 0.000 description 1
- 229910000027 potassium carbonate Inorganic materials 0.000 description 1
- 230000017854 proteolysis Effects 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- OHRURASPPZQGQM-GCCNXGTGSA-N romidepsin Chemical compound O1C(=O)[C@H](C(C)C)NC(=O)C(=C/C)/NC(=O)[C@H]2CSSCC\C=C\[C@@H]1CC(=O)N[C@H](C(C)C)C(=O)N2 OHRURASPPZQGQM-GCCNXGTGSA-N 0.000 description 1
- IZTQOLKUZKXIRV-YRVFCXMDSA-N sincalide Chemical compound C([C@@H](C(=O)N[C@@H](CCSC)C(=O)NCC(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC=1C=CC=CC=1)C(N)=O)NC(=O)[C@@H](N)CC(O)=O)C1=CC=C(OS(O)(=O)=O)C=C1 IZTQOLKUZKXIRV-YRVFCXMDSA-N 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- FPGGTKZVZWFYPV-UHFFFAOYSA-M tetrabutylammonium fluoride Chemical compound [F-].CCCC[N+](CCCC)(CCCC)CCCC FPGGTKZVZWFYPV-UHFFFAOYSA-M 0.000 description 1
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000002103 transcriptional effect Effects 0.000 description 1
- 229910000404 tripotassium phosphate Inorganic materials 0.000 description 1
- 235000019798 tripotassium phosphate Nutrition 0.000 description 1
- 230000034512 ubiquitination Effects 0.000 description 1
- 238000010798 ubiquitination Methods 0.000 description 1
- 238000011144 upstream manufacturing Methods 0.000 description 1
- WAEXFXRVDQXREF-UHFFFAOYSA-N vorinostat Chemical compound ONC(=O)CCCCCCC(=O)NC1=CC=CC=C1 WAEXFXRVDQXREF-UHFFFAOYSA-N 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D495/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having sulfur atoms as the only ring hetero atoms
- C07D495/12—Heterocyclic compounds containing in the condensed system at least one hetero ring having sulfur atoms as the only ring hetero atoms in which the condensed system contains three hetero rings
- C07D495/14—Ortho-condensed systems
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
Abstract
本发明公开了一种BETHDAC双靶向PROTAC的制备方法和应用，化合物具有如通式(I)的结构。该类化合物是通过BET抑制剂((+)‑JQ1)与HDAC抑制剂和E3泛素连接酶进行合理设计而来。药理实验表明这类化合物对BET蛋白与HDAC蛋白具有较强的结合抑制活性，以及体外抗肿瘤增殖活性。机制实验表明，化合物BET/HDAC双靶PROTAC能够明显的诱导BRD4蛋白与HDACs蛋白进行降解，可以应用于BRD4与HDACs蛋白介导的病理学特征的肿瘤疾病。该类化合物作为首次报道的BET/HDAC双靶PROTAC分子，具有进一步的开发和研究价值。
Description
技术领域
本发明涉及医药技术领域，具体为一种BETHDAC双靶向PROTAC的制备方法和应用，以及该类小分子化合物在治疗恶性肿瘤疾病中的应用。
背景技术
PROTAC作为一种新兴的技术，通过双功能小分子使靶蛋白被E3泛素连接酶识别并打上泛素化标签，然后利用细胞自身的泛素-蛋白酶体系统降解靶蛋白。目前，大多数有关PROTAC的研究都集中在降解单个靶标蛋白上。尽管有一些文献中已经发表了可以实现两种或多种蛋白的下调，但大多数是由PROTAC脱靶效应引起的，或者是由于抑制了上游信号引起下游多种蛋白下调的结果，或由于作用于相同类型蛋白的配体口袋导致，因此PROTAC分子可以同时实现下调多个相同类型的靶蛋白，例如目前报道对激酶的降解。目前，尚未实现使用PROTAC技术将不同类型的靶蛋白进行同步双重敲除。这方面的主要挑战是不同类型蛋白质的配体结合口袋非常不同，并且设计可同时结合不同类型靶蛋白的配体更加困难。其次，因为PROTAC化合物是一种双功能分子介导的三元复合物，复合物形成和识别的动态过程涉及E3连接酶与靶蛋白之间的相互作用，因此对双功能分子本身的大小和结构都有严格的要求。利用小分子选择性地使不同类型的靶蛋白进行同步协同敲除，不仅将产生具有发展潜力的治疗方法，而且还将提供有价值的工具，进行化学和生物学相关的研究。
BET家族在人体组织广泛表达，与多种疾病的发生和发展密切相关，近年来研究发现，BET蛋白参与睾丸核蛋白中线癌(NUT midline carcinoma，NMC)、急性白血病、淋巴瘤、乳腺癌等肿瘤的发生、浸润和转移过程。BET家族成员BRD4可调节癌基因MYC，BCL2和BCL6等转录过程。抑制或敲除BRD4，可诱发肿瘤细胞凋亡，从而达到抗肿瘤作用；组蛋白去乙酰化酶(HDACs)是一种重要的表观遗传学清除剂，可从组蛋白的赖氨酸上除去乙酰基。它们包含11个组分，分为4类(I，IIa，IIb和IV)和7个沉默调节蛋白(III类)。在癌细胞中，HDACs的过度表达导致去乙酰化作用增强，不利于特定基因的表达，包括一些肿瘤抑制基因。迄今为止，已经批准了五种HDAC抑制剂(HDACis)，即伏立诺他(SAHA)，罗米地辛(FK228)，贝利诺司他(PXD-101)，帕诺比司他(LBH-589)和甲胺酰胺，用于治疗血液癌和胰腺癌。BRD4与HDACs这两个靶标在部分肿瘤的发生和发展中都起着重要作用，均是癌症和其他慢性疾病中的表观遗传学靶标，已有研究报道同时抑制BRD4和HDACs可以协同下调c-Myc等癌基因的表达，达到协同抗肿瘤的效果。本课题组前期通过药效团融合策略设计合成了一系列新颖的BET/HDACs双靶抑制剂，在体外与体内都展现了出色的抗肿瘤效果。但这类双靶抑制剂仍具有一般抑制剂的某些局限性，例如严重的脱靶毒性和更高剂量的依赖性。从理论上说，使用PROTAC实现BRD4和HDACs的双重敲除将克服双靶抑制剂所存在的问题，目前尚无文献报道使用合理设计的PROTAC同时对两种靶蛋白的双重敲除。基于上述原因，我们使用PROTAC技术设计出了能够将BRD4和HDACs进行同步敲除的双靶PROTAC分子。
发明内容
本发明的目的是提供一类基于BRD4蛋白与HDACs蛋白降解剂及其药学上可接受的盐。本发明还提供该化合物的制备方法和应用。
本发明的技术方案如下：
1、通式(I)的化合物及药学上可接受的盐：
X为1至60个碳原子的饱和或不饱和的直链烃基、氧杂链、间氧杂链、苯基、杂环基团或以下链接基团之一：
Y为1至60个碳原子的饱和或不饱和的直链烃基、氧杂链、间氧杂链、苯基、杂环基团或以下链接基团之一：
其中n是0-30。
其中，优选的杂环基团为哌嗪基、吡咯基、吡唑基、呋喃基、噻吩基、噁唑基、异噁唑基、噻唑基、异噻唑基、吡啶基、嘧啶基、吡嗪基或哒嗪基或以下链接基团之一：
其中n是0-30。
其中，优选的杂环基团为哌嗪酮基、吡咯基、吡唑基、呋喃基、噻吩基、噁唑基、异噁唑基、噻唑基、异噻唑基、吡啶基、嘧啶基、吡嗪基或哒嗪基。A或B为卤素、烯烃、醇、酚、醚、醛、酮、羧酸、硝基、磺酸类有机物、胺类、酰胺类、酯基或下列基团：-X、-OH、-CHO、-COOH、-NO2、-SO3H、-NH2、-CO-
其中n是0-30。
所述的药学上可接受的盐包括通式I化合物与下列酸形成的酸加成盐：盐酸、氢溴酸、硫酸、乳酸、柠檬酸、磷酸、甲磺酸、苯磺酸、对甲苯磺酸、萘磺酸、酒石酸、丙酮酸、乙酸、马来酸或琥珀酸、富马酸、水杨酸、苯基乙酸、杏仁酸。
根据本发明，进一步优选的，上述通式(I)化合物是下列之一：
8-(2-((S)-4-(4-氯苯基)-2，3，9-三甲基-6H-噻吩并[3，2-f][1，2，4]三唑[4，3-a][1，4]二氮杂-1-基)-N-(2-(2-(2-(2-((2-(2，6-二氧代哌啶-3-基)1-氧代异吲哚-4-基)氨基)-2-氧乙氧基)乙氧基)乙氧基)乙基)乙酰胺基)-N-羟基辛酰胺(M10)
8-(2-((S)-4-(4-氯苯基)-2,3,9-三甲基-6H-噻吩并[3,2-f][1,2,4]三唑[4,3-a][1,4]二氮杂-6-6基)-N-(2-(2-(2-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚-4-基]氨基)-2-氧代乙氧基)乙氧基)乙基)乙酰氨基)辛酸(M12)
8-(2-((S)-4-(4-氯苯基)-2,3,9-三甲基-6H-噻吩并[3,2-f][1,2,4]三唑[4,3-a][1,4]二氮杂-1-基)-N-(2-(3-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚-4-基)氨基)-3-氧代丙氧基)乙基)乙酰胺基)辛酸(M14)
14-(2-((S)-4-(4-氯苯基)-2,3,9-三甲基-6H-噻吩并[3,2-f]
[1,2,4]三唑[4,3-a][1,4]二氮杂-6-6基)-N-(8-(羟基氨基)-8-氧辛基)乙酰氨基)-N-(2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-4-基)-369,,,12-四氧杂环丁酸酯(M16)
17-(2-((S)-4-(4-氯苯基)-2,3,9-三甲基-6H-噻吩并[3,2-f]
[1,2,4]三唑[4,3-a][1,4]二氮杂-6-6基)-N-(8-(羟基氨基)-8-氧辛基)乙酰氨基)-N-(2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-4-基)-369,,,12,15-五氧杂十八碳酰胺(M18)
20-(2-((S)-4-(4-氯苯基)-2,3,9-三甲基-6H-噻吩并[3,2-f]
[1,2,4]三唑[4,3-a][1,4]二氮杂-6-6基)-N-(8-(羟基氨基)-8-氧辛基)乙酰氨基)-N-(2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-4-基)-369,,,12,15,18-六氧杂二十二酰胺(M20)
以上优选化合物，后面的括号内的编号是对应于下面反应路线及表中的化合物结构的编号。
本发明的化合物制备方法如下：
通法：
通法：
5，8，11-三氧杂-2-氮杂十三烷二酸-1-叔丁酯(M0)与来那度胺(M1)在DMF中进行缩合反应得到化合物M2，
M2在三氟乙酸存在的条件下脱去Boc保护基得到化合物M3，M3与M4进行取代反应得到化合物M5,
M5与(+)-JQ1羧酸进行缩合反应得到化合物M6,
M6在碱性条件下进行酯交换反应得到M7，
M7在三氟乙酸条件下脱去羧基保护基叔丁酯得到化合物M8,
M8与O-三苯甲基羟胺进行缩合反应得到化合物M11，
M11最后在三氟乙酸存在的条件下脱去保护基暴露出异羟肟酸基团得到化合物M10。
其中HATU为2-(7-氧化苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯，DIPEA为N，N-二异丙基乙胺，MeOH为甲醇，CH2Cl2为二氯甲烷，DMF为二甲基甲酰胺，TEA为三乙胺以下含义类同。
化合物M1与其他原料通过市场购买；
本发明也提供了上述结构通式(I)所示化合物在制备BRD4降抑制剂或HDACs抑制剂中的应用。
本发明还提供了上述结构通式(I)所示化合物在治疗与上述靶点有关的肿瘤疾病中的应用，包括对肿瘤细胞的杀伤作用。与BRD4或HDACs有关的疾病可以是，但不限于：肺癌、肝癌、肾癌、非小细胞肺癌、前列腺癌、甲状腺癌、皮肤癌、胰腺癌、卵巢癌、乳腺癌、膀胱癌、骨髓增生异常综合症、淋巴瘤、食管癌、胃肠道癌、骨肉瘤、中枢或外周神经系统的肿瘤。
附图说明
图1为本发明提出的一种BETHDAC双靶向PROTAC的制备方法和应用的分析结果示意图。
具体实施方式
现结合实施例，对本发明作详细描述，但本发明的实施不仅限于此。本发明所用试剂和原料均市售可得或可按文献方法制备。
下列实施例中未注明具体条件的实验方法，通常按照常规条件，或按照制造厂商所建议的条件。
以下实施例所涉化合物的1HNMR，13CNMR和MS数据详见表1.1。
表1.1优选化合物1HNMR，13CNMR和MS数据
实施例1：
8-(2-((S)-4-(4-氯苯基)-2，3，9-三甲基-6H-噻吩并[3，2-f][1，2，4]三唑[4，3-a][1，4]二氮杂-1-基)-N-(2-(2-(2-(2-((2-(2，6-二氧代哌啶-3-基)1-氧代异吲哚-4-基)氨基)-2-氧乙氧基)乙氧基)乙氧基)乙基)乙酰胺基)-N-羟基辛酰胺的合成(M10)
步骤a：叔丁基(2-(2-(2-(2-(2-((2-(2，6-二氧代哌啶-3-基)1-氧代异吲哚基-4-基)氨基)-2-氧代乙氧基)乙氧基)乙氧基)氨基甲酸乙酯的合成(M2)
步骤a：将化合物M0(1.0g,3.1mmol)H，ATU(2.4g,6.2mmol)D，IPEA(1.0mL,7.8mmol)溶于干燥的DMF中，在室温条件下反应0.5h然后加入来那度胺M1(0.80g,3.1mmol)后反应4h，经TLC检测反应完全。将反应液缓慢倒入冰水(1.5L)混合物中，用EA(30mL×3)萃取后合并有机相，减压蒸除溶剂，剩余物用硅胶柱色谱分离，流动相为DCM/MeOH＝100/5,得到黄色油状液体M2(1.2g,2.2mmol,产率：70％)。
步骤b：2-(2-(2-(2-(2-氨基乙氧基)乙氧基)乙氧基)-N-(2-(2，6-二氧代哌啶-3-基)-1-氧代异吲哚-4-基)乙酰胺的合成(M3)
步骤b：将化合物M2(600mg,1.1mmol)溶于干燥的二氯甲烷(8.0mL)中加入TFA(2.0mL)在室温条件下反应2h后经TLC检测反应较完全，减压蒸除溶剂，剩余物无需处理直接投下一步反应。
步骤c：1-((2-(2，6-二氧代哌啶-3-基)-1-氧代异吲哚-4-基)氨基)-1-氧代-3，6，9-三氧杂-12-氮杂二十烷酸乙酯的合成(M5)
将化合物M4(203mg,0.81mmol)，K2CO3(224mg,1.6mmol)，KI(6.7mg,0.041mmol)溶于干燥的乙腈中，在40℃条件下反应十分钟后加入化合物M3(363mg,0.81mmol)反应12h，经TLC检测反应完全。将溶剂减压蒸除，剩余物用硅胶柱色谱分离，流动相为DCM/MeOH＝100/7,得到无色透明液体M5(150mg,0.24mmol,产率：30％)。
步骤d：乙基12-(2-((S)-4-(4-氯苯基)-2，3，9-三甲基-6H-噻吩并[3，2-f][1，2，4]三唑并[4，3-a][1，4]二氮杂卓-6-基)乙酰基)-1-((2-(2，6-二氧哌啶-3-基)-1-氧代异吲哚-4-基)氨基)-1-氧代-3，6，9-三氧代-12-氮杂-20-酸盐的合成(M6)
步骤d：将化合物(+)-JQ1羧酸(200mg,0.50mmol)，HATU(380mg,1.0mmol)，DIPEA(162mg,1.3mmol)加入到干燥的DMF中，在室温条件下反应0.5h后加入化合物M4(309mg,0.5mmol)反应4h，经TLC检测反应完全。将反应液缓慢倒入冰水(500mL)混合物中，用EA(20mL×3)萃取后合并有机相，减压蒸除溶剂，剩余物用硅胶柱色谱分离流动相为DCM/MeOH＝100/3,得到白色固体M6(500mg,0.40mmol,产率：80％)。
步骤e：叔丁基12-(2-((S)-4-(4-氯苯基)-2，3，9-三甲基-6H-噻吩并[3，2-f][1，2，4]三唑并[4，3-a][1，4]二氮杂-6-基)乙酰基)-1-((2-(2，6-二氧哌啶-3-基)-1-氧代异吲哚-4-基)氨基)-1-氧代-3，6，9-三氧代-12-氮杂-20-酸盐的合成(M7)
步骤e：将化合物M5(100mg,0.080mmol)，TBAF(21mg,0.080mmol)LiOH(7.6mg,0.32mmol)，溶于干燥的四氢呋喃中，在80℃条件下反应2h后，加入化合物叔丁醇(30mg,0.40mmol)反应4h，经TLC检测反应较完全，减压蒸除溶剂，剩余物用硅胶柱色谱分离，流动相为DCM/MeOH＝100/5,得到白色固体M7(58mg,0.056mmol,产率：70％)。
步骤f：12-(2-((S)-4-(4-氯苯基)-2，3，9-三甲基-6H-噻吩并[3，2-f][1，2，4]三唑并[4，3-a][1，4]二氮杂卓-6-基)乙酰基)-1-((2-(2，6-二氧哌啶-3-基)-1-氧代异吲哚-4-基)氨基)-1-氧代-3，6，9-三氧代-12-氮杂-20-甲酸的合成(M8)
步骤f：将化合物M7(100mg,0.097mmol)溶于干燥的二氯甲烷(8.0mL)中，加入TFA(2.0mL)在室温条件下反应4h，经TLC检测反应较完全，减压蒸除溶剂，剩余物无需处理直接投下一步反应。
步骤g：8-(2-((S)-4-(4-氯苯基)-2，3，9-三甲基-6H-噻吩并[3，2-f][1，2，4]三唑[4，3-a][1，4]二氮杂-1-基)-N-(2-(2-(2-(2-((2-(2，6-二氧代哌啶-3-基)1-氧代异吲哚-4-基)氨基)-2-氧代乙氧基)乙氧基)乙氧基)乙基)乙酰胺基)-N-(三苯甲氧基)辛酰胺的合成(M11)
步骤g：将化合物M8(50mg,0.051mmol)，HATU(39mg,0.102mmol)，DIPEA(16mg,0.13mmol)溶于干燥的DMF中，在室温条件下反应0.5h后，加入化合物O-三苯甲基羟胺(72mg,0.26mmol)反应2h，经TLC检测反应完全，将反应液缓慢倒入冰水(30mL)混合物中用EA(10mL×3)萃取后合并有机相，减压蒸除溶剂，剩余物用硅胶柱色谱分离，流动相为DCM/MeOH＝100/5,得到白色固体M11(53mg,0.043mmol,产率：85％)。
步骤h：8-(2-((S)-4-(4-氯苯基)-2，3，9-三甲基-6H-噻吩并[3，2-f][1，2，4]三唑[4，3-a][1，4]二氮杂-1-基)-N-(2-(2-(2-(2-((2-(2，6-二氧代哌啶-3-基)1-氧代异吲哚-4-基)氨基)-2-氧乙氧基)乙氧基)乙氧基)乙基)乙酰胺基)-N-羟基辛酰胺的合成(M10)
步骤h：将化合物M11(53mg,0.043mmol)溶于干燥的二氯甲烷(8.0mL)中，然后加入TFA(2.0mL)在室温条件下反应3h，经TLC检测反应完全，减压蒸除溶剂，剩余物用硅胶柱色谱分离，流动相为DCM/MeOH＝100/10,得到白色固体M10(39mg,0.039mmol,产率：90％)。
化合物M12,M14,M16,M18,M20的合成过程参照化合物M10的合成步骤。
实施例2：本发明化合物对BRD4与HDAC1抑制活性测试(Ki)。
将5μL待测化合物(各稀释浓度)、BRD4(20nM)及PMDM6-F(20nM)(缓冲液：100mM磷酸三钾，PH＝7.5；100μg/mL BGG；0.02％叠氮钠)加入96孔黑色平底酶标板至终体积115μL，室温孵育1小时后，用Biotek-Synergy酶标仪(激发光为485nM，发射光为528nM)读数荧光偏振值。
依据上述方法得到的荧光偏振值，用Origin 9.0软件绘制曲线，计算得蛋白结合抑制常数(Ki)。HDAC1测试方法同BRD4。
实验结果：首先测试所有目标化合物对BRD4和HDAC1蛋白的抑制活性，选用(+)-JQ1与SAHA作为阳性对照。测试结果如表4-2所示，保留与BRD4和HDACs蛋白结合活性基团的化合物M10对两种蛋白均有较强的抑制活性；对与BRD4产生结合作用的基团进行封闭只保留与HDACs产生结合作用的基团，化合物M12展现出对HDAC1的较强抑制活性，对BRD4没有抑制活性；对与HDACs产生结合作用的基团进行封闭只保留与BRD4产生结合作用的基团，化合物M11展现出对BRD4的较强抑制活性，对HDAC1没有抑制活性；对与BRD4和HDACs都产生结合作用的基团进行封闭的化合物M13对两种蛋白都没有抑制活性。通过封闭能够与靶蛋白产生结合的活性基团，初步验证了BET/HDAC双靶PROTAC化合物M10对两种靶蛋白都能产生抑制活性，说明M10能够与靶蛋白上的活性位点结合，这是PROTAC分子发挥降解功能的基础与预期结果相符合。
表1.2目标化合物对BRD4和HDAC1的抑制活性(IC50,μM)
实施例3：本发明化合物的体外抗肿瘤活性测试(IC50)。
对本发明的化合物进行了三种肿瘤细胞增殖抑制能力测试，测试方法采用常规的CCK8法。将对于对数生长期的肿瘤细胞(MCF-7(人乳腺癌细胞)、A549(人肺癌细胞)、HepG2(人肝癌细胞)用胰酶消化，然后用培养基(DMEM+10％FBS或者PRMI1640+10％FBS)将细胞稀释悬浮成单细胞悬液，调整细胞密度为5×104个/mL，每孔加入100μL接种于96孔板中，置37℃、5％CO2培养箱内培养24小时，再分别加入不同浓度的化合物，每个浓度平行三个复孔，并设置实验组和对照组，继续孵育72小时后，向每孔加入10μLCCK8溶液，然后37℃下避光孵育1-4小时后，用Biotek-Synergy酶标仪测450nm OD值。计算半数抑制浓度IC50。
实验结果：进一步评价目标化合物的体外抗肿瘤活性，选用A549(人肺癌细胞)、HepG2(人肝癌细胞)、MCF-7(人乳腺癌细胞)作为测试肿瘤株，(+)-JQ1与SAHA作为阳性对照。测试结果如表4-3所示，整体看化合物M10在所有目标化合物中抗肿瘤活性最好比阳性药稍好；M13在所有化合物中抗肿瘤活性最差；M11与M12抗肿瘤活性介于M10与M13之间，目标化合物的体外抗肿瘤活性与化合物对BRD4和HDAC1抑酶活性基本保持一致，抑酶活性与细胞活性结果均显示，化合物M10在所有目标化合物中表现出最好的活性，达到化合物的设计要求，下面对目标化合物开展作用机制研究，进一步验证BET/HDAC双靶PROTAC的设计思想。
表1.3.目标化合物的体外抗肿瘤活性(IC50,μM)a
Claims (5)
1.一种BETHDAC双靶向PROTAC的制备方法和应用，包括，其特征在于，通式(I)的化合物及它们药学上可接受的盐：
X为1至60个碳原子的饱和或不饱和的直链烃基、氧杂链、间氧杂链、苯基、杂环基团或以下链接基团之一：
Y为1至60个碳原子的饱和或不饱和的直链烃基、氧杂链、间氧杂链、苯基、杂环基团或以下链接基团之一：
其中n＝0-30。
其中，优选的杂环基团为哌嗪基、吡咯基、吡唑基、呋喃基、噻吩基、噁唑基、异噁唑基、噻唑基、异噻唑基、吡啶基、嘧啶基、吡嗪基或哒嗪基或以下链接基团之一：
其中n是0-30。
其中，优选的杂环基团为哌嗪酮基、吡咯基、吡唑基、呋喃基、噻吩基、噁唑基、异噁唑基、噻唑基、异噻唑基、吡啶基、嘧啶基、吡嗪基或哒嗪基。
A或B为卤素、烯烃、醇、酚、醚、醛、酮、羧酸、硝基、磺酸类有机物、胺类、酰胺类、酯基或下列基团：-X、-OH、-CHO、-COOH、-NO2、-SO3H、-NH2、-CO-
其中n是0-30。
2.权利要求1的上述结构通式(I)所示化合物，其特征在如下化合物：
8-(2-((S)-4-(4-氯苯基)-2，3，9-三甲基-6H-噻吩并[3，2-f][1，2，4]三唑[4，3-a][1，4]二氮杂-1-基)-N-(2-(2-(2-(2-((2-(2，6-二氧代哌啶-3-基)1-氧代异吲哚-4-基)氨基)-2-氧乙氧基)乙氧基)乙氧基)乙基)乙酰胺基)-N-羟基辛酰胺(1)
8-(2-((S)-4-(4-氯苯基)-2,3,9-三甲基-6H-噻吩并[3,2-f][1,2,4]三唑[4,3-a][1,4]二氮杂-6-6基)-N-(2-(2-(2-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚-4-基]氨基)-2-氧代乙氧基)乙氧基)乙基)乙酰氨基)辛酸(2)
8-(2-((S)-4-(4-氯苯基)-2,3,9-三甲基-6H-噻吩并[3,2-f][1,2,4]三唑[4,3-a][1,4]二氮杂-1-基)-N-(2-(3-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚-4-基)氨基)-3-氧代丙氧基)乙基)乙酰胺基)辛酸(3)
14-(2-((S)-4-(4-氯苯基)-2,3,9-三甲基-6H-噻吩并[3,2-f][1,2,4]三唑[4,3-a][1,4]二氮杂-6-6基)-N-(8-(羟基氨基)-8-氧辛基)乙酰氨基)-N-(2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-4-基)-3,6,9,12-四氧杂环丁酸酯(4)
17-(2-((S)-4-(4-氯苯基)-2,3,9-三甲基-6H-噻吩并[3,2-f][1,2,4]三唑[4,3-a][1,4]二氮杂-6-6基)-N-(8-(羟基氨基)-8-氧辛基)乙酰氨基)-N-(2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-4-基)-3,6,9,12,15-五氧杂十八碳酰胺(5)
20-(2-((S)-4-(4-氯苯基)-2,3,9-三甲基-6H-噻吩并[3,2-f][1,2,4]三唑[4,3-a][1,4]二氮杂-6-6基)-N-(8-(羟基氨基)-8-氧辛基)乙酰氨基)-N-(2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-4-基)-3,6,9,12,15,18-六氧杂二十二酰胺(6)。
3.如权利要求1的上述结构通式(I)所示化合物，其药学上可接受的盐包括通式(I)化合物与下列酸形成的酸加成盐：盐酸、氢溴酸、硫酸、乳酸、柠檬酸、磷酸、甲磺酸、苯磺酸、对甲苯磺酸、萘磺酸、酒石酸、丙酮酸、乙酸、马来酸或琥珀酸、富马酸、水杨酸、苯基乙酸、杏仁酸。
4.如权利要求1的上述结构通式(I)所示化合物在制备BET抑制剂与HDAC抑制剂中的应用。
5.如权利要求1上述结构通式(I)所示化合物在治疗与上述靶点有关的肿瘤疾病中的应用，包括对肿瘤细胞的杀伤作用，以及在裸鼠肿瘤模型中延缓肿瘤生长的应用。与BRD4蛋白和HDAC蛋白有关的疾病可以是，但不限于：肺癌、肝癌、肾癌、非小细胞肺癌、前列腺癌、甲状腺癌、皮肤癌、胰腺癌、卵巢癌、乳腺癌、膀胱癌、骨髓增生异常综合症、淋巴瘤、食管癌、胃肠道癌、骨肉瘤、中枢或外周神经系统的肿瘤。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110627242.3A CN113651835A (zh) | 2021-06-04 | 2021-06-04 | 一种bethdac双靶向protac的制备方法和应用 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110627242.3A CN113651835A (zh) | 2021-06-04 | 2021-06-04 | 一种bethdac双靶向protac的制备方法和应用 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN113651835A true CN113651835A (zh) | 2021-11-16 |
Family
ID=78489117
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110627242.3A Pending CN113651835A (zh) | 2021-06-04 | 2021-06-04 | 一种bethdac双靶向protac的制备方法和应用 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113651835A (zh) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114890989A (zh) * | 2022-05-25 | 2022-08-12 | 佛山市晨康生物科技有限公司 | 一种含氮衍生物为Linker的HDAC8降解剂其制备方法和应用 |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20190233433A1 (en) * | 2018-01-26 | 2019-08-01 | Yale University | Imide-based modulators of proteolysis and associated methods of use |
| US20190255066A1 (en) * | 2016-09-14 | 2019-08-22 | University Of Dundee | Fluorohydroxyproline derivatives useful in the preparation of proteolysis targeted chimeras |
| US20190275161A1 (en) * | 2016-06-10 | 2019-09-12 | Otsuka Pharmaceutical Co., Ltd. | Cliptac composition |
| CN114106010A (zh) * | 2021-04-20 | 2022-03-01 | 上海大学 | 一种蛋白降解靶向嵌合体的制备方法和应用 |
-
2021
- 2021-06-04 CN CN202110627242.3A patent/CN113651835A/zh active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20190275161A1 (en) * | 2016-06-10 | 2019-09-12 | Otsuka Pharmaceutical Co., Ltd. | Cliptac composition |
| US20190255066A1 (en) * | 2016-09-14 | 2019-08-22 | University Of Dundee | Fluorohydroxyproline derivatives useful in the preparation of proteolysis targeted chimeras |
| US20190233433A1 (en) * | 2018-01-26 | 2019-08-01 | Yale University | Imide-based modulators of proteolysis and associated methods of use |
| CN114106010A (zh) * | 2021-04-20 | 2022-03-01 | 上海大学 | 一种蛋白降解靶向嵌合体的制备方法和应用 |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114890989A (zh) * | 2022-05-25 | 2022-08-12 | 佛山市晨康生物科技有限公司 | 一种含氮衍生物为Linker的HDAC8降解剂其制备方法和应用 |
| CN114890989B (zh) * | 2022-05-25 | 2024-03-22 | 广东晨康生物科技有限公司 | 一种含氮衍生物为Linker的HDAC8降解剂其制备方法和应用 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Girardini et al. | Cereblon versus VHL: Hijacking E3 ligases against each other using PROTACs | |
| CN114106010A (zh) | 一种蛋白降解靶向嵌合体的制备方法和应用 | |
| JP2020516650A (ja) | 小分子 | |
| Shinji et al. | Design and synthesis of phthalimide-type histone deacetylase inhibitors | |
| A Salvador et al. | Discovery and mechanism of natural products as modulators of histone acetylation | |
| Baud et al. | Thioester derivatives of the natural product psammaplin A as potent histone deacetylase inhibitors | |
| Cai et al. | Novel hybrids from N-hydroxyarylamide and indole ring through click chemistry as histone deacetylase inhibitors with potent antitumor activities | |
| Kim et al. | An overview of naturally occurring histone deacetylase inhibitors | |
| Ghosh et al. | Dissecting structure–activity-relationships of crebinostat: Brain penetrant HDAC inhibitors for neuroepigenetic regulation | |
| CN113651835A (zh) | 一种bethdac双靶向protac的制备方法和应用 | |
| Liu et al. | Synthesis and biological evaluation of paclitaxel and vorinostat co-prodrugs for overcoming drug resistance in cancer therapy in vitro | |
| Lee et al. | Design, synthesis, and evaluation of isoindolinone-hydroxamic acid derivatives as histone deacetylase (HDAC) inhibitors | |
| Li et al. | Synthesis, antitumor activity evaluation and mechanistic study of novel hederacolchiside A1 derivatives bearing an aryl triazole moiety | |
| EP3684754A1 (en) | Pentafluorophenyl sulfonamide compounds, compositions and uses thereof | |
| CN103420923B (zh) | 4-氨基喹唑啉异羟肟酸类化合物及作为抗肿瘤药物应用 | |
| Schäker‐Hübner et al. | Balancing Histone Deacetylase (HDAC) Inhibition and Drug‐likeness: Biological and Physicochemical Evaluation of Class I Selective HDAC Inhibitors | |
| Kawamura et al. | Design and synthesis of the stabilized analogs of belactosin A with the unnatural cis-cyclopropane structure | |
| KR100456047B1 (ko) | 애피시딘 유도체, 이의 합성방법 및 이를 포함하는 항암제조성물 | |
| Oliveira et al. | Synthesis and pharmacological evaluation of novel isoquinoline N-sulphonylhydrazones designed as ROCK inhibitors | |
| Kojić et al. | Structure based design, synthesis and in vitro antitumour activity of tiazofurin stereoisomers with nitrogen functions at the C-2′ or C-3′ positions | |
| Yang et al. | Enantioselective semisynthesis of novel cephalotaxine esters with potent antineoplastic activities against leukemia | |
| Arsianti et al. | Synthesis and anticancer activity of polyhydroxylated 18-membered analogue of antimycin A3 | |
| Šimková et al. | Structure-activity relationship and biochemical evaluation of novel fibroblast activation protein and prolyl endopeptidase inhibitors with α-ketoamide warheads | |
| Zhao et al. | A rational design strategy of the novel topoisomerase II inhibitors for the synthesis of the 4-O-(2-pyrazinecarboxylic)-4′-demethylepipodophyllotoxin with antitumor activity by diminishing the relaxation reaction of topoisomerase II-DNA decatenation | |
| Wang et al. | Synthesis and Antimetastatic Effects of E‐Salignone Amide Derivatives |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20211116 |
|
| RJ01 | Rejection of invention patent application after publication |