CN109897036B - Triazolopyridine compounds and preparation method and use thereof - Google Patents
Triazolopyridine compounds and preparation method and use thereof Download PDFInfo
- Publication number
- CN109897036B CN109897036B CN201910200914.5A CN201910200914A CN109897036B CN 109897036 B CN109897036 B CN 109897036B CN 201910200914 A CN201910200914 A CN 201910200914A CN 109897036 B CN109897036 B CN 109897036B
- Authority
- CN
- China
- Prior art keywords
- alkyl
- pharmaceutically acceptable
- stereoisomers
- acceptable salts
- methyl
- 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.)
- Active
Links
- 150000008523 triazolopyridines Chemical class 0.000 title claims abstract description 26
- 238000002360 preparation method Methods 0.000 title abstract description 13
- 150000001875 compounds Chemical class 0.000 claims abstract description 30
- 150000003839 salts Chemical class 0.000 claims abstract description 29
- 102000008096 B7-H1 Antigen Human genes 0.000 claims abstract description 23
- 108010074708 B7-H1 Antigen Proteins 0.000 claims abstract description 23
- 206010028980 Neoplasm Diseases 0.000 claims abstract description 11
- 201000010099 disease Diseases 0.000 claims abstract description 9
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims abstract description 9
- 201000011510 cancer Diseases 0.000 claims abstract description 7
- 239000003814 drug Substances 0.000 claims abstract description 7
- 230000019491 signal transduction Effects 0.000 claims abstract description 7
- 230000009385 viral infection Effects 0.000 claims abstract description 3
- 125000000217 alkyl group Chemical group 0.000 claims description 66
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 claims description 19
- 125000002887 hydroxy group Chemical group [H]O* 0.000 claims description 17
- 229910052736 halogen Inorganic materials 0.000 claims description 16
- 150000002367 halogens Chemical class 0.000 claims description 16
- 229910052739 hydrogen Inorganic materials 0.000 claims description 15
- 239000001257 hydrogen Substances 0.000 claims description 15
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 claims description 12
- 125000004397 aminosulfonyl group Chemical group NS(=O)(=O)* 0.000 claims description 11
- 125000003917 carbamoyl group Chemical group [H]N([H])C(*)=O 0.000 claims description 11
- 208000023275 Autoimmune disease Diseases 0.000 claims description 9
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 claims description 9
- 150000002431 hydrogen Chemical class 0.000 claims description 8
- 229910052757 nitrogen Inorganic materials 0.000 claims description 8
- 125000004433 nitrogen atom Chemical group N* 0.000 claims description 8
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 claims description 7
- 125000004093 cyano group Chemical group *C#N 0.000 claims description 6
- 125000000623 heterocyclic group Chemical group 0.000 claims description 6
- QJGQUHMNIGDVPM-UHFFFAOYSA-N nitrogen group Chemical group [N] QJGQUHMNIGDVPM-UHFFFAOYSA-N 0.000 claims description 6
- 208000035473 Communicable disease Diseases 0.000 claims description 5
- 239000000460 chlorine Substances 0.000 claims description 5
- 229910052801 chlorine Inorganic materials 0.000 claims description 5
- ZAMOUSCENKQFHK-UHFFFAOYSA-N Chlorine atom Chemical compound [Cl] ZAMOUSCENKQFHK-UHFFFAOYSA-N 0.000 claims description 4
- 208000018359 Systemic autoimmune disease Diseases 0.000 claims description 4
- 210000000056 organ Anatomy 0.000 claims description 4
- 239000008194 pharmaceutical composition Substances 0.000 claims description 4
- 239000004480 active ingredient Substances 0.000 claims description 3
- 238000004519 manufacturing process Methods 0.000 claims description 3
- 201000001441 melanoma Diseases 0.000 claims description 3
- 125000000956 methoxy group Chemical group [H]C([H])([H])O* 0.000 claims description 3
- 206010061311 nervous system neoplasm Diseases 0.000 claims description 3
- 239000000546 pharmaceutical excipient Substances 0.000 claims description 3
- 208000035143 Bacterial infection Diseases 0.000 claims description 2
- 206010006187 Breast cancer Diseases 0.000 claims description 2
- 208000026310 Breast neoplasm Diseases 0.000 claims description 2
- 206010009900 Colitis ulcerative Diseases 0.000 claims description 2
- 208000002699 Digestive System Neoplasms Diseases 0.000 claims description 2
- 208000032612 Glial tumor Diseases 0.000 claims description 2
- 206010018338 Glioma Diseases 0.000 claims description 2
- 208000001204 Hashimoto Disease Diseases 0.000 claims description 2
- 208000030836 Hashimoto thyroiditis Diseases 0.000 claims description 2
- 208000035186 Hemolytic Autoimmune Anemia Diseases 0.000 claims description 2
- 206010020850 Hyperthyroidism Diseases 0.000 claims description 2
- 206010058467 Lung neoplasm malignant Diseases 0.000 claims description 2
- 206010025323 Lymphomas Diseases 0.000 claims description 2
- 206010029240 Neuritis Diseases 0.000 claims description 2
- 206010036105 Polyneuropathy Diseases 0.000 claims description 2
- 208000000453 Skin Neoplasms Diseases 0.000 claims description 2
- 208000004732 Systemic Vasculitis Diseases 0.000 claims description 2
- 206010067584 Type 1 diabetes mellitus Diseases 0.000 claims description 2
- 201000006704 Ulcerative Colitis Diseases 0.000 claims description 2
- 206010047115 Vasculitis Diseases 0.000 claims description 2
- 208000036142 Viral infection Diseases 0.000 claims description 2
- 230000001154 acute effect Effects 0.000 claims description 2
- 201000000448 autoimmune hemolytic anemia Diseases 0.000 claims description 2
- 208000022362 bacterial infectious disease Diseases 0.000 claims description 2
- 239000003937 drug carrier Substances 0.000 claims description 2
- 208000015181 infectious disease Diseases 0.000 claims description 2
- 201000005202 lung cancer Diseases 0.000 claims description 2
- 208000020816 lung neoplasm Diseases 0.000 claims description 2
- 208000019629 polyneuritis Diseases 0.000 claims description 2
- 230000002265 prevention Effects 0.000 claims description 2
- 206010039073 rheumatoid arthritis Diseases 0.000 claims description 2
- 201000000849 skin cancer Diseases 0.000 claims description 2
- 201000000596 systemic lupus erythematosus Diseases 0.000 claims description 2
- 208000035408 type 1 diabetes mellitus 1 Diseases 0.000 claims description 2
- 125000000753 cycloalkyl group Chemical group 0.000 claims 8
- HNQIVZYLYMDVSB-UHFFFAOYSA-N methanesulfonimidic acid Chemical group CS(N)(=O)=O HNQIVZYLYMDVSB-UHFFFAOYSA-N 0.000 claims 8
- 238000006467 substitution reaction Methods 0.000 claims 8
- 125000003545 alkoxy group Chemical group 0.000 claims 3
- 201000005787 hematologic cancer Diseases 0.000 claims 1
- 208000024200 hematopoietic and lymphoid system neoplasm Diseases 0.000 claims 1
- 201000011682 nervous system cancer Diseases 0.000 claims 1
- -1 triazolopyridine compound Chemical class 0.000 abstract description 94
- 230000002401 inhibitory effect Effects 0.000 abstract description 5
- 239000000203 mixture Substances 0.000 abstract description 5
- 230000006916 protein interaction Effects 0.000 abstract description 5
- 229940079593 drug Drugs 0.000 abstract description 3
- 102100040678 Programmed cell death protein 1 Human genes 0.000 abstract 2
- 101710089372 Programmed cell death protein 1 Proteins 0.000 abstract 2
- 238000006243 chemical reaction Methods 0.000 description 58
- 125000004178 (C1-C4) alkyl group Chemical group 0.000 description 40
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 39
- 238000002330 electrospray ionisation mass spectrometry Methods 0.000 description 38
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 33
- 239000007787 solid Substances 0.000 description 31
- 239000000243 solution Substances 0.000 description 21
- 101100519207 Mus musculus Pdcd1 gene Proteins 0.000 description 20
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 20
- 238000004440 column chromatography Methods 0.000 description 19
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 18
- DPGSPRJLAZGUBQ-UHFFFAOYSA-N 4,4,5,5-tetramethyl-2-vinyl-1,3,2-dioxaborolane Substances CC1(C)OB(C=C)OC1(C)C DPGSPRJLAZGUBQ-UHFFFAOYSA-N 0.000 description 16
- JQWHASGSAFIOCM-UHFFFAOYSA-M sodium periodate Chemical compound [Na+].[O-]I(=O)(=O)=O JQWHASGSAFIOCM-UHFFFAOYSA-M 0.000 description 16
- 239000011259 mixed solution Substances 0.000 description 15
- RYHBNJHYFVUHQT-UHFFFAOYSA-N 1,4-Dioxane Chemical compound C1COCCO1 RYHBNJHYFVUHQT-UHFFFAOYSA-N 0.000 description 12
- HZAXFHJVJLSVMW-UHFFFAOYSA-N 2-Aminoethan-1-ol Chemical compound NCCO HZAXFHJVJLSVMW-UHFFFAOYSA-N 0.000 description 12
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 12
- 239000002994 raw material Substances 0.000 description 11
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 10
- 238000000034 method Methods 0.000 description 10
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 10
- 238000000967 suction filtration Methods 0.000 description 10
- UHOVQNZJYSORNB-UHFFFAOYSA-N Benzene Chemical compound C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 description 8
- JGFZNNIVVJXRND-UHFFFAOYSA-N N,N-Diisopropylethylamine (DIPEA) Chemical compound CCN(C(C)C)C(C)C JGFZNNIVVJXRND-UHFFFAOYSA-N 0.000 description 8
- NEZGSPBXJRTLBH-UHFFFAOYSA-N OC(=O)c1cc(Br)cn2cnnc12 Chemical compound OC(=O)c1cc(Br)cn2cnnc12 NEZGSPBXJRTLBH-UHFFFAOYSA-N 0.000 description 8
- HXITXNWTGFUOAU-UHFFFAOYSA-N phenylboronic acid Chemical compound OB(O)C1=CC=CC=C1 HXITXNWTGFUOAU-UHFFFAOYSA-N 0.000 description 8
- BWHMMNNQKKPAPP-UHFFFAOYSA-L potassium carbonate Chemical compound [K+].[K+].[O-]C([O-])=O BWHMMNNQKKPAPP-UHFFFAOYSA-L 0.000 description 8
- BEOOHQFXGBMRKU-UHFFFAOYSA-N sodium cyanoborohydride Chemical compound [Na+].[B-]C#N BEOOHQFXGBMRKU-UHFFFAOYSA-N 0.000 description 8
- 125000000229 (C1-C4)alkoxy group Chemical group 0.000 description 7
- 239000012044 organic layer Substances 0.000 description 7
- 125000006552 (C3-C8) cycloalkyl group Chemical group 0.000 description 6
- BABPEUPHRVFJIJ-UHFFFAOYSA-N 6-[(2-hydroxyethylamino)methyl]-N-(2-methyl-3-phenylphenyl)-[1,2,4]triazolo[4,3-a]pyridine-8-carboxamide Chemical compound OCCNCC=1C=C(C=2N(C=1)C=NN=2)C(=O)NC=1C(=C(C=CC=1)C1=CC=CC=C1)C BABPEUPHRVFJIJ-UHFFFAOYSA-N 0.000 description 6
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 6
- 125000004453 alkoxycarbonyl group Chemical group 0.000 description 6
- 239000007864 aqueous solution Substances 0.000 description 6
- JFDZBHWFFUWGJE-UHFFFAOYSA-N benzonitrile Chemical compound N#CC1=CC=CC=C1 JFDZBHWFFUWGJE-UHFFFAOYSA-N 0.000 description 6
- 239000013024 dilution buffer Substances 0.000 description 6
- 230000000694 effects Effects 0.000 description 6
- 239000012321 sodium triacetoxyborohydride Substances 0.000 description 6
- IILVSKMKMOJHMA-UHFFFAOYSA-N 3-bromo-2-methylaniline Chemical compound CC1=C(N)C=CC=C1Br IILVSKMKMOJHMA-UHFFFAOYSA-N 0.000 description 5
- ZCYVEMRRCGMTRW-UHFFFAOYSA-N 7553-56-2 Chemical compound [I] ZCYVEMRRCGMTRW-UHFFFAOYSA-N 0.000 description 5
- 229960000583 acetic acid Drugs 0.000 description 5
- GDTBXPJZTBHREO-UHFFFAOYSA-N bromine Substances BrBr GDTBXPJZTBHREO-UHFFFAOYSA-N 0.000 description 5
- 229910052794 bromium Inorganic materials 0.000 description 5
- FJDQFPXHSGXQBY-UHFFFAOYSA-L caesium carbonate Chemical compound [Cs+].[Cs+].[O-]C([O-])=O FJDQFPXHSGXQBY-UHFFFAOYSA-L 0.000 description 5
- 229910000024 caesium carbonate Inorganic materials 0.000 description 5
- NXQGGXCHGDYOHB-UHFFFAOYSA-L cyclopenta-1,4-dien-1-yl(diphenyl)phosphane;dichloropalladium;iron(2+) Chemical compound [Fe+2].Cl[Pd]Cl.[CH-]1C=CC(P(C=2C=CC=CC=2)C=2C=CC=CC=2)=C1.[CH-]1C=CC(P(C=2C=CC=CC=2)C=2C=CC=CC=2)=C1 NXQGGXCHGDYOHB-UHFFFAOYSA-L 0.000 description 5
- 239000012065 filter cake Substances 0.000 description 5
- 239000000706 filtrate Substances 0.000 description 5
- 239000012362 glacial acetic acid Substances 0.000 description 5
- 230000003993 interaction Effects 0.000 description 5
- 239000011630 iodine Substances 0.000 description 5
- 229910052740 iodine Inorganic materials 0.000 description 5
- 239000002904 solvent Substances 0.000 description 5
- 238000010189 synthetic method Methods 0.000 description 5
- 239000007821 HATU Substances 0.000 description 4
- 229920002472 Starch Polymers 0.000 description 4
- 238000006069 Suzuki reaction reaction Methods 0.000 description 4
- 210000001744 T-lymphocyte Anatomy 0.000 description 4
- 238000001514 detection method Methods 0.000 description 4
- 229910000027 potassium carbonate Inorganic materials 0.000 description 4
- 239000008107 starch Substances 0.000 description 4
- 235000019698 starch Nutrition 0.000 description 4
- 238000003756 stirring Methods 0.000 description 4
- PYOKUURKVVELLB-UHFFFAOYSA-N trimethyl orthoformate Chemical compound COC(OC)OC PYOKUURKVVELLB-UHFFFAOYSA-N 0.000 description 4
- 210000004881 tumor cell Anatomy 0.000 description 4
- 125000004939 6-pyridyl group Chemical group N1=CC=CC=C1* 0.000 description 3
- WKBOTKDWSSQWDR-UHFFFAOYSA-N Bromine atom Chemical compound [Br] WKBOTKDWSSQWDR-UHFFFAOYSA-N 0.000 description 3
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 3
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 3
- 239000012269 PD-1/PD-L1 inhibitor Substances 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
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 description 3
- 125000002252 acyl group Chemical group 0.000 description 3
- 229960003767 alanine Drugs 0.000 description 3
- 238000007112 amidation reaction Methods 0.000 description 3
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 3
- 238000011161 development Methods 0.000 description 3
- 238000009169 immunotherapy Methods 0.000 description 3
- 230000003647 oxidation Effects 0.000 description 3
- 238000007254 oxidation reaction Methods 0.000 description 3
- YJVFFLUZDVXJQI-UHFFFAOYSA-L palladium(ii) acetate Chemical compound [Pd+2].CC([O-])=O.CC([O-])=O YJVFFLUZDVXJQI-UHFFFAOYSA-L 0.000 description 3
- 229940121653 pd-1/pd-l1 inhibitor Drugs 0.000 description 3
- 150000003384 small molecules Chemical class 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
- WIIBAHCYWOUFRM-UHFFFAOYSA-N 2-(2,3-dihydro-1,4-benzodioxin-6-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane Chemical compound O1C(C)(C)C(C)(C)OB1C1=CC=C(OCCO2)C2=C1 WIIBAHCYWOUFRM-UHFFFAOYSA-N 0.000 description 2
- ZKZYTSGUONPOPF-UHFFFAOYSA-N 2-amino-6-bromobenzonitrile Chemical compound NC1=CC=CC(Br)=C1C#N ZKZYTSGUONPOPF-UHFFFAOYSA-N 0.000 description 2
- LMAIDQDTPOQJBC-UHFFFAOYSA-N 2-amino-6-phenylbenzonitrile Chemical compound NC1=CC=CC(C=2C=CC=CC=2)=C1C#N LMAIDQDTPOQJBC-UHFFFAOYSA-N 0.000 description 2
- WXVGZIZBMFJASZ-UHFFFAOYSA-N 2-methyl-3-phenylaniline Chemical compound CC1=C(N)C=CC=C1C1=CC=CC=C1 WXVGZIZBMFJASZ-UHFFFAOYSA-N 0.000 description 2
- HTDYGZQDPPNVLL-UHFFFAOYSA-N 6-[(2-acetamidoethylamino)methyl]-N-[3-(2,3-dihydro-1,4-benzodioxin-6-yl)-2-methylphenyl]-[1,2,4]triazolo[4,3-a]pyridine-8-carboxamide Chemical compound C(C)(=O)NCCNCC=1C=C(C=2N(C=1)C=NN=2)C(=O)NC1=C(C(=CC=C1)C1=CC2=C(OCCO2)C=C1)C HTDYGZQDPPNVLL-UHFFFAOYSA-N 0.000 description 2
- SSWKJNCEBUNBPU-UHFFFAOYSA-N 6-[[2,3-dihydroxypropyl(methyl)amino]methyl]-N-(2-methyl-3-phenylphenyl)-[1,2,4]triazolo[4,3-a]pyridine-8-carboxamide Chemical compound OC(CN(C)CC=1C=C(C=2N(C=1)C=NN=2)C(=O)NC=1C(=C(C=CC=1)C1=CC=CC=C1)C)CO SSWKJNCEBUNBPU-UHFFFAOYSA-N 0.000 description 2
- ZEKRPGVBVNLJFQ-UHFFFAOYSA-N 6-[[2-hydroxyethyl(methyl)amino]methyl]-N-(2-methyl-3-phenylphenyl)-[1,2,4]triazolo[4,3-a]pyridine-8-carboxamide Chemical compound OCCN(C)CC=1C=C(C=2N(C=1)C=NN=2)C(=O)NC=1C(=C(C=CC=1)C1=CC=CC=C1)C ZEKRPGVBVNLJFQ-UHFFFAOYSA-N 0.000 description 2
- UYVDHCNWWSSFSK-UHFFFAOYSA-N 6-bromo-N-(2-methyl-3-phenylphenyl)-[1,2,4]triazolo[4,3-a]pyridine-8-carboxamide Chemical compound BrC=1C=C(C=2N(C=1)C=NN=2)C(=O)NC=1C(=C(C=CC=1)C1=CC=CC=C1)C UYVDHCNWWSSFSK-UHFFFAOYSA-N 0.000 description 2
- NKTREENIJKJCEC-UHFFFAOYSA-N 6-formyl-N-(2-methyl-3-phenylphenyl)-[1,2,4]triazolo[4,3-a]pyridine-8-carboxamide Chemical compound C(=O)C=1C=C(C=2N(C=1)C=NN=2)C(=O)NC=1C(=C(C=CC=1)C1=CC=CC=C1)C NKTREENIJKJCEC-UHFFFAOYSA-N 0.000 description 2
- GDBLUSGIMVGCSA-HZCBDIJESA-N CC(C(C1=CC=CC=C1)=CC=C1)=C1NC(C1=CC(CN[C@H](CC2)CC[C@@H]2O)=CN2C1=NN=C2)=O Chemical compound CC(C(C1=CC=CC=C1)=CC=C1)=C1NC(C1=CC(CN[C@H](CC2)CC[C@@H]2O)=CN2C1=NN=C2)=O GDBLUSGIMVGCSA-HZCBDIJESA-N 0.000 description 2
- FYXCHRIDCPSENN-UHFFFAOYSA-N COC(=O)c1cc(Br)cn2cnnc12 Chemical compound COC(=O)c1cc(Br)cn2cnnc12 FYXCHRIDCPSENN-UHFFFAOYSA-N 0.000 description 2
- XHROVBRSNFGKAP-UHFFFAOYSA-N COC(=O)c1cc(Br)cnc1NN Chemical compound COC(=O)c1cc(Br)cnc1NN XHROVBRSNFGKAP-UHFFFAOYSA-N 0.000 description 2
- 239000004375 Dextrin Substances 0.000 description 2
- 229920001353 Dextrin Polymers 0.000 description 2
- KRHYYFGTRYWZRS-UHFFFAOYSA-N Fluorane Chemical compound F KRHYYFGTRYWZRS-UHFFFAOYSA-N 0.000 description 2
- VZCYOOQTPOCHFL-OWOJBTEDSA-N Fumaric acid Chemical compound OC(=O)\C=C\C(O)=O VZCYOOQTPOCHFL-OWOJBTEDSA-N 0.000 description 2
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 2
- 108010074328 Interferon-gamma Proteins 0.000 description 2
- 102000000588 Interleukin-2 Human genes 0.000 description 2
- 108010002350 Interleukin-2 Proteins 0.000 description 2
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 2
- AFVFQIVMOAPDHO-UHFFFAOYSA-N Methanesulfonic acid Chemical compound CS(O)(=O)=O AFVFQIVMOAPDHO-UHFFFAOYSA-N 0.000 description 2
- CISNFTBPQKKIBA-UHFFFAOYSA-N N-(2-methyl-3-phenylphenyl)-6-[(2-sulfamoylethylamino)methyl]-[1,2,4]triazolo[4,3-a]pyridine-8-carboxamide Chemical compound CC1=C(C=CC=C1NC(=O)C=1C=2N(C=C(C=1)CNCCS(N)(=O)=O)C=NN=2)C1=CC=CC=C1 CISNFTBPQKKIBA-UHFFFAOYSA-N 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- 239000002202 Polyethylene glycol Substances 0.000 description 2
- 102000007056 Recombinant Fusion Proteins Human genes 0.000 description 2
- 108010008281 Recombinant Fusion Proteins Proteins 0.000 description 2
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- 229910001413 alkali metal ion Inorganic materials 0.000 description 2
- 230000009435 amidation Effects 0.000 description 2
- 125000003118 aryl group Chemical group 0.000 description 2
- 239000011230 binding agent Substances 0.000 description 2
- 125000001246 bromo group Chemical group Br* 0.000 description 2
- 239000000872 buffer Substances 0.000 description 2
- 230000003915 cell function Effects 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 230000008878 coupling Effects 0.000 description 2
- 238000010168 coupling process Methods 0.000 description 2
- 238000005859 coupling reaction Methods 0.000 description 2
- 235000019425 dextrin Nutrition 0.000 description 2
- 239000003085 diluting agent Substances 0.000 description 2
- 238000001035 drying Methods 0.000 description 2
- 239000008103 glucose Substances 0.000 description 2
- IKDUDTNKRLTJSI-UHFFFAOYSA-N hydrazine monohydrate Substances O.NN IKDUDTNKRLTJSI-UHFFFAOYSA-N 0.000 description 2
- 238000002347 injection Methods 0.000 description 2
- 239000007924 injection Substances 0.000 description 2
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 2
- 239000000314 lubricant Substances 0.000 description 2
- MOMQDEDQGJAKII-UHFFFAOYSA-N methyl 5-bromo-2-chloropyridine-3-carboxylate Chemical compound COC(=O)C1=CC(Br)=CN=C1Cl MOMQDEDQGJAKII-UHFFFAOYSA-N 0.000 description 2
- 150000007522 mineralic acids Chemical class 0.000 description 2
- 150000007524 organic acids Chemical class 0.000 description 2
- 229910000489 osmium tetroxide Inorganic materials 0.000 description 2
- NFHFRUOZVGFOOS-UHFFFAOYSA-N palladium;triphenylphosphane Chemical compound [Pd].C1=CC=CC=C1P(C=1C=CC=CC=1)C1=CC=CC=C1.C1=CC=CC=C1P(C=1C=CC=CC=1)C1=CC=CC=C1.C1=CC=CC=C1P(C=1C=CC=CC=1)C1=CC=CC=C1.C1=CC=CC=C1P(C=1C=CC=CC=1)C1=CC=CC=C1 NFHFRUOZVGFOOS-UHFFFAOYSA-N 0.000 description 2
- 229920001223 polyethylene glycol Polymers 0.000 description 2
- 239000000651 prodrug Substances 0.000 description 2
- 229940002612 prodrug Drugs 0.000 description 2
- 238000006268 reductive amination reaction Methods 0.000 description 2
- 239000002002 slurry Substances 0.000 description 2
- 239000011734 sodium Substances 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- FYSNRJHAOHDILO-UHFFFAOYSA-N thionyl chloride Chemical compound ClS(Cl)=O FYSNRJHAOHDILO-UHFFFAOYSA-N 0.000 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 description 2
- VZCYOOQTPOCHFL-UHFFFAOYSA-N trans-butenedioic acid Natural products OC(=O)C=CC(O)=O VZCYOOQTPOCHFL-UHFFFAOYSA-N 0.000 description 2
- 239000000080 wetting agent Substances 0.000 description 2
- BJEPYKJPYRNKOW-REOHCLBHSA-N (S)-malic acid Chemical compound OC(=O)[C@@H](O)CC(O)=O BJEPYKJPYRNKOW-REOHCLBHSA-N 0.000 description 1
- NGNBDVOYPDDBFK-UHFFFAOYSA-N 2-[2,4-di(pentan-2-yl)phenoxy]acetyl chloride Chemical compound CCCC(C)C1=CC=C(OCC(Cl)=O)C(C(C)CCC)=C1 NGNBDVOYPDDBFK-UHFFFAOYSA-N 0.000 description 1
- LUMPMFJBIPSKQV-UHFFFAOYSA-N 3-(2,3-dihydro-1,4-benzodioxin-6-yl)-2-methylaniline Chemical compound CC1=C(N)C=CC=C1C1=CC=C2OCCOC2=C1 LUMPMFJBIPSKQV-UHFFFAOYSA-N 0.000 description 1
- BMYNFMYTOJXKLE-UHFFFAOYSA-N 3-azaniumyl-2-hydroxypropanoate Chemical compound NCC(O)C(O)=O BMYNFMYTOJXKLE-UHFFFAOYSA-N 0.000 description 1
- HKNLHCGTRMCOLV-UHFFFAOYSA-N 3-bromo-2-chloroaniline Chemical compound NC1=CC=CC(Br)=C1Cl HKNLHCGTRMCOLV-UHFFFAOYSA-N 0.000 description 1
- CMEFWSXWVZBBJP-UHFFFAOYSA-N 6-bromo-N-[3-(2,3-dihydro-1,4-benzodioxin-6-yl)-2-methylphenyl]-[1,2,4]triazolo[4,3-a]pyridine-8-carboxamide Chemical compound BrC=1C=C(C=2N(C=1)C=NN=2)C(=O)NC1=C(C(=CC=C1)C1=CC2=C(OCCO2)C=C1)C CMEFWSXWVZBBJP-UHFFFAOYSA-N 0.000 description 1
- 241000220479 Acacia Species 0.000 description 1
- GUBGYTABKSRVRQ-XLOQQCSPSA-N Alpha-Lactose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@H](O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-N 0.000 description 1
- 101100463133 Caenorhabditis elegans pdl-1 gene Proteins 0.000 description 1
- 102000000844 Cell Surface Receptors Human genes 0.000 description 1
- 108010001857 Cell Surface Receptors Proteins 0.000 description 1
- 229920002785 Croscarmellose sodium Polymers 0.000 description 1
- 108090000695 Cytokines Proteins 0.000 description 1
- 102000004127 Cytokines Human genes 0.000 description 1
- FBPFZTCFMRRESA-FSIIMWSLSA-N D-Glucitol Natural products OC[C@H](O)[C@H](O)[C@@H](O)[C@H](O)CO FBPFZTCFMRRESA-FSIIMWSLSA-N 0.000 description 1
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 1
- FBPFZTCFMRRESA-JGWLITMVSA-N D-glucitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-JGWLITMVSA-N 0.000 description 1
- FEWJPZIEWOKRBE-JCYAYHJZSA-N Dextrotartaric acid Chemical compound OC(=O)[C@H](O)[C@@H](O)C(O)=O FEWJPZIEWOKRBE-JCYAYHJZSA-N 0.000 description 1
- 239000001856 Ethyl cellulose Substances 0.000 description 1
- ZZSNKZQZMQGXPY-UHFFFAOYSA-N Ethyl cellulose Chemical compound CCOCC1OC(OC)C(OCC)C(OCC)C1OC1C(O)C(O)C(OC)C(CO)O1 ZZSNKZQZMQGXPY-UHFFFAOYSA-N 0.000 description 1
- PXGOKWXKJXAPGV-UHFFFAOYSA-N Fluorine Chemical compound FF PXGOKWXKJXAPGV-UHFFFAOYSA-N 0.000 description 1
- 108010010803 Gelatin Proteins 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
- 102100037850 Interferon gamma Human genes 0.000 description 1
- 102000008070 Interferon-gamma Human genes 0.000 description 1
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 1
- 235000010643 Leucaena leucocephala Nutrition 0.000 description 1
- WHXSMMKQMYFTQS-UHFFFAOYSA-N Lithium Chemical compound [Li] WHXSMMKQMYFTQS-UHFFFAOYSA-N 0.000 description 1
- 229930195725 Mannitol Natural products 0.000 description 1
- 229920000168 Microcrystalline cellulose Polymers 0.000 description 1
- 229920000715 Mucilage Polymers 0.000 description 1
- HSHXDCVZWHOWCS-UHFFFAOYSA-N N'-hexadecylthiophene-2-carbohydrazide Chemical compound CCCCCCCCCCCCCCCCNNC(=O)c1cccs1 HSHXDCVZWHOWCS-UHFFFAOYSA-N 0.000 description 1
- FBOMDXUFPHBCPI-UHFFFAOYSA-N N-[3-(2,3-dihydro-1,4-benzodioxin-6-yl)-2-methylphenyl]-6-formyl-[1,2,4]triazolo[4,3-a]pyridine-8-carboxamide Chemical compound O1C2=C(OCC1)C=C(C=C2)C=1C(=C(C=CC=1)NC(=O)C=1C=2N(C=C(C=1)C=O)C=NN=2)C FBOMDXUFPHBCPI-UHFFFAOYSA-N 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- OFOBLEOULBTSOW-UHFFFAOYSA-N Propanedioic acid Natural products OC(=O)CC(O)=O OFOBLEOULBTSOW-UHFFFAOYSA-N 0.000 description 1
- 239000004365 Protease Substances 0.000 description 1
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 1
- 208000005718 Stomach Neoplasms Diseases 0.000 description 1
- KDYFGRWQOYBRFD-UHFFFAOYSA-N Succinic acid Natural products OC(=O)CCC(O)=O KDYFGRWQOYBRFD-UHFFFAOYSA-N 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 230000006044 T cell activation Effects 0.000 description 1
- 230000006052 T cell proliferation Effects 0.000 description 1
- FEWJPZIEWOKRBE-UHFFFAOYSA-N Tartaric acid Natural products [H+].[H+].[O-]C(=O)C(O)C(O)C([O-])=O FEWJPZIEWOKRBE-UHFFFAOYSA-N 0.000 description 1
- TVXBFESIOXBWNM-UHFFFAOYSA-N Xylitol Natural products OCCC(O)C(O)C(O)CCO TVXBFESIOXBWNM-UHFFFAOYSA-N 0.000 description 1
- DPXJVFZANSGRMM-UHFFFAOYSA-N acetic acid;2,3,4,5,6-pentahydroxyhexanal;sodium Chemical compound [Na].CC(O)=O.OCC(O)C(O)C(O)C(O)C=O DPXJVFZANSGRMM-UHFFFAOYSA-N 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 239000000853 adhesive Substances 0.000 description 1
- BJEPYKJPYRNKOW-UHFFFAOYSA-N alpha-hydroxysuccinic acid Natural products OC(=O)C(O)CC(O)=O BJEPYKJPYRNKOW-UHFFFAOYSA-N 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- 150000001448 anilines Chemical class 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 229960003852 atezolizumab Drugs 0.000 description 1
- 229950002916 avelumab Drugs 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 150000001555 benzenes Chemical class 0.000 description 1
- HFACYLZERDEVSX-UHFFFAOYSA-N benzidine Chemical class C1=CC(N)=CC=C1C1=CC=C(N)C=C1 HFACYLZERDEVSX-UHFFFAOYSA-N 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 230000000903 blocking effect Effects 0.000 description 1
- ZADPBFCGQRWHPN-UHFFFAOYSA-N boronic acid Chemical compound OBO ZADPBFCGQRWHPN-UHFFFAOYSA-N 0.000 description 1
- KDYFGRWQOYBRFD-NUQCWPJISA-N butanedioic acid Chemical compound O[14C](=O)CC[14C](O)=O KDYFGRWQOYBRFD-NUQCWPJISA-N 0.000 description 1
- FUFJGUQYACFECW-UHFFFAOYSA-L calcium hydrogenphosphate Chemical compound [Ca+2].OP([O-])([O-])=O FUFJGUQYACFECW-UHFFFAOYSA-L 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- UHKPOGGUWJGGID-UHFFFAOYSA-N carbonic acid;cesium Chemical compound [Cs].OC(O)=O UHKPOGGUWJGGID-UHFFFAOYSA-N 0.000 description 1
- 239000001768 carboxy methyl cellulose Substances 0.000 description 1
- 150000001732 carboxylic acid derivatives Chemical class 0.000 description 1
- 239000000969 carrier Substances 0.000 description 1
- 238000007385 chemical modification Methods 0.000 description 1
- 125000001309 chloro group Chemical group Cl* 0.000 description 1
- 235000015165 citric acid Nutrition 0.000 description 1
- 239000001767 crosslinked sodium carboxy methyl cellulose Substances 0.000 description 1
- 235000010947 crosslinked sodium carboxy methyl cellulose Nutrition 0.000 description 1
- 239000012043 crude product Substances 0.000 description 1
- 125000004122 cyclic group Chemical group 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 235000019700 dicalcium phosphate Nutrition 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- GPAYUJZHTULNBE-UHFFFAOYSA-O diphenylphosphanium Chemical compound C=1C=CC=CC=1[PH2+]C1=CC=CC=C1 GPAYUJZHTULNBE-UHFFFAOYSA-O 0.000 description 1
- 239000007884 disintegrant Substances 0.000 description 1
- 239000002552 dosage form Substances 0.000 description 1
- 229950009791 durvalumab Drugs 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- CCIVGXIOQKPBKL-UHFFFAOYSA-M ethanesulfonate Chemical compound CCS([O-])(=O)=O CCIVGXIOQKPBKL-UHFFFAOYSA-M 0.000 description 1
- 235000019325 ethyl cellulose Nutrition 0.000 description 1
- 229920001249 ethyl cellulose Polymers 0.000 description 1
- 230000017188 evasion or tolerance of host immune response Effects 0.000 description 1
- 230000005284 excitation Effects 0.000 description 1
- 238000002474 experimental method Methods 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
- 229910052731 fluorine Inorganic materials 0.000 description 1
- 239000011737 fluorine Substances 0.000 description 1
- 239000001530 fumaric acid Substances 0.000 description 1
- 206010017758 gastric cancer Diseases 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 235000001727 glucose Nutrition 0.000 description 1
- 229940093915 gynecological organic acid Drugs 0.000 description 1
- 208000019691 hematopoietic and lymphoid cell neoplasm Diseases 0.000 description 1
- 125000001072 heteroaryl group Chemical group 0.000 description 1
- 235000012907 honey Nutrition 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 239000001866 hydroxypropyl methyl cellulose Substances 0.000 description 1
- 235000010979 hydroxypropyl methyl cellulose Nutrition 0.000 description 1
- 229920003088 hydroxypropyl methyl cellulose Polymers 0.000 description 1
- 230000002163 immunogen Effects 0.000 description 1
- 230000004957 immunoregulator effect Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 229960003130 interferon gamma Drugs 0.000 description 1
- HXEACLLIILLPRG-RXMQYKEDSA-N l-pipecolic acid Natural products OC(=O)[C@H]1CCCCN1 HXEACLLIILLPRG-RXMQYKEDSA-N 0.000 description 1
- 239000004310 lactic acid Substances 0.000 description 1
- 235000014655 lactic acid Nutrition 0.000 description 1
- 239000008101 lactose Substances 0.000 description 1
- 229910001416 lithium ion Inorganic materials 0.000 description 1
- 229940031703 low substituted hydroxypropyl cellulose Drugs 0.000 description 1
- VZCYOOQTPOCHFL-UPHRSURJSA-N maleic acid Chemical compound OC(=O)\C=C/C(O)=O VZCYOOQTPOCHFL-UPHRSURJSA-N 0.000 description 1
- 239000011976 maleic acid Substances 0.000 description 1
- 239000001630 malic acid Substances 0.000 description 1
- 235000011090 malic acid Nutrition 0.000 description 1
- 239000000594 mannitol Substances 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- HEBKCHPVOIAQTA-UHFFFAOYSA-N meso ribitol Natural products OCC(O)C(O)C(O)CO HEBKCHPVOIAQTA-UHFFFAOYSA-N 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 229940098779 methanesulfonic acid Drugs 0.000 description 1
- 235000019813 microcrystalline cellulose Nutrition 0.000 description 1
- 239000008108 microcrystalline cellulose Substances 0.000 description 1
- 229940016286 microcrystalline cellulose Drugs 0.000 description 1
- 229940125645 monoclonal antibody drug Drugs 0.000 description 1
- 125000002950 monocyclic group Chemical group 0.000 description 1
- 229960003301 nivolumab Drugs 0.000 description 1
- 208000002154 non-small cell lung carcinoma Diseases 0.000 description 1
- 238000005457 optimization Methods 0.000 description 1
- 235000005985 organic acids Nutrition 0.000 description 1
- PIBWKRNGBLPSSY-UHFFFAOYSA-L palladium(II) chloride Chemical compound Cl[Pd]Cl PIBWKRNGBLPSSY-UHFFFAOYSA-L 0.000 description 1
- 229960002621 pembrolizumab Drugs 0.000 description 1
- 230000004962 physiological condition Effects 0.000 description 1
- HXEACLLIILLPRG-UHFFFAOYSA-N pipecolic acid Chemical compound OC(=O)C1CCCCN1 HXEACLLIILLPRG-UHFFFAOYSA-N 0.000 description 1
- 125000003367 polycyclic group Chemical group 0.000 description 1
- 239000001267 polyvinylpyrrolidone Substances 0.000 description 1
- 229920000036 polyvinylpyrrolidone Polymers 0.000 description 1
- 235000013855 polyvinylpyrrolidone Nutrition 0.000 description 1
- 230000008092 positive effect Effects 0.000 description 1
- 229910001414 potassium ion Inorganic materials 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 238000010992 reflux Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000013207 serial dilution Methods 0.000 description 1
- 230000001568 sexual effect Effects 0.000 description 1
- 239000000377 silicon dioxide Substances 0.000 description 1
- 235000012239 silicon dioxide Nutrition 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 235000015424 sodium Nutrition 0.000 description 1
- 235000019812 sodium carboxymethyl cellulose Nutrition 0.000 description 1
- 229920001027 sodium carboxymethylcellulose Polymers 0.000 description 1
- 229910001415 sodium ion Inorganic materials 0.000 description 1
- DAJSVUQLFFJUSX-UHFFFAOYSA-M sodium;dodecane-1-sulfonate Chemical compound [Na+].CCCCCCCCCCCCS([O-])(=O)=O DAJSVUQLFFJUSX-UHFFFAOYSA-M 0.000 description 1
- 238000003797 solvolysis reaction Methods 0.000 description 1
- 239000000600 sorbitol Substances 0.000 description 1
- 235000010356 sorbitol Nutrition 0.000 description 1
- 229940032147 starch Drugs 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 201000011549 stomach cancer Diseases 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 235000020357 syrup Nutrition 0.000 description 1
- 239000006188 syrup Substances 0.000 description 1
- 239000003826 tablet Substances 0.000 description 1
- 239000000454 talc Substances 0.000 description 1
- 229910052623 talc Inorganic materials 0.000 description 1
- 230000008685 targeting Effects 0.000 description 1
- 239000011975 tartaric acid Substances 0.000 description 1
- 235000002906 tartaric acid Nutrition 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 206010044412 transitional cell carcinoma Diseases 0.000 description 1
- OVCXRBARSPBVMC-UHFFFAOYSA-N triazolopyridine Chemical compound C=1N2C(C(C)C)=NN=C2C=CC=1C=1OC=NC=1C1=CC=C(F)C=C1 OVCXRBARSPBVMC-UHFFFAOYSA-N 0.000 description 1
- 208000029729 tumor suppressor gene on chromosome 11 Diseases 0.000 description 1
- 230000003827 upregulation Effects 0.000 description 1
- 239000000811 xylitol Substances 0.000 description 1
- 235000010447 xylitol Nutrition 0.000 description 1
- HEBKCHPVOIAQTA-SCDXWVJYSA-N xylitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)CO HEBKCHPVOIAQTA-SCDXWVJYSA-N 0.000 description 1
- 229960002675 xylitol Drugs 0.000 description 1
Landscapes
- Nitrogen Condensed Heterocyclic Rings (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
本发明属于医药技术领域，涉及三唑并吡啶类化合物及其制备方法和用途。具体涉及具有通式Ⅰ结构的三唑并吡啶类化合物及其立体异构体以及药学上可接受的盐，其中R1、R2、R3、R4如权利要求和说明书所述。本发明所述的化合物及其立体异构体以及药学上可接受的盐和含有该化合物的组合物均对PD‑1/PD‑L1蛋白/蛋白相互作用具有显著抑制作用，能够治疗癌症、病毒感染等多种疾病，因此，可用于制备预防和/或治疗与PD‑1/PD‑L1信号通路有关的疾病的药物。
Description
技术领域：Technical field:
本发明属于医药技术领域，涉及三唑并吡啶类化合物及其立体异构体以及药学上可接受的盐，它们的制备方法以及含有所述化合物的药物组合物。本发明还涉及该类化合物及其立体异构体以及药学上可接受的盐在制备治疗与PD-1/PD-L1信号通路有关的疾病如癌症、感染性疾病、自身免疫性疾病的药物方面的用途。The invention belongs to the technical field of medicine, and relates to triazolopyridine compounds, stereoisomers and pharmaceutically acceptable salts, their preparation methods and pharmaceutical compositions containing the compounds. The present invention also relates to the compounds and their stereoisomers and pharmaceutically acceptable salts in the preparation of medicines for treating diseases related to PD-1/PD-L1 signaling pathway, such as cancer, infectious diseases and autoimmune diseases the use of.
背景技术：Background technique:
免疫治疗是近年来肿瘤治疗的热点领域，于2013年被《Science》评为十大科学突破之首。程序性死亡受体1(programmed death 1，PD-1)是T细胞表面受体，当其与程序性死亡配体1(PD-L1)结合时产生负性免疫调节信号，从而抑制T细胞活化、增殖以及白细胞介素2(IL-2)、干扰素γ(IFN-γ)等细胞因子的释放(Eur.J.Immunol.,2002,32(3),634-643.)。大量研究表明，机体内的肿瘤微环境会诱导浸润的T细胞中PD-1表达上调，同时肿瘤细胞高表达PD-L1，导致PD-1/PD-L1介导的信号通路持续激活，肿瘤特异性CD8+T细胞功能被抑制以至于不能识别或杀伤肿瘤细胞，即肿瘤细胞实现免疫逃逸。因此靶向阻断PD-1/PD-L1蛋白/蛋白相互作用，可以恢复T细胞功能，使其重新识别并杀伤肿瘤细胞。Immunotherapy is a hot field of tumor treatment in recent years, and was rated as the top ten scientific breakthroughs by "Science" in 2013. Programmed death 1 (PD-1) is a T cell surface receptor that when it binds to programmed death ligand 1 (PD-L1) produces a negative immunoregulatory signal, thereby inhibiting T cell activation , proliferation and the release of cytokines such as interleukin 2 (IL-2) and interferon gamma (IFN-γ) (Eur. J. Immunol., 2002, 32(3), 634-643.). A large number of studies have shown that the tumor microenvironment in the body can induce the up-regulation of PD-1 expression in infiltrating T cells, and at the same time, tumor cells highly express PD-L1, resulting in the continuous activation of PD-1/PD-L1-mediated signaling pathways, and tumor specificity. Sexual CD8+ T cell function is suppressed so that it cannot recognize or kill tumor cells, that is, tumor cells achieve immune escape. Therefore, targeted blocking of PD-1/PD-L1 protein/protein interaction can restore T cell function, allowing it to re-recognize and kill tumor cells.
基于PD-1/PD-L1的免疫疗法备受瞩目，目前已被批准上市的PD-1/PD-L1单抗包括默沙东的Pembrolizumab、百时美施贵宝的Nivolumab、默克的Avelumab、阿斯利康的Durvalumab、罗氏的Atezolizumab等。上述单抗已在多种肿瘤类型的治疗中显示出明显疗效，被批准的适应症包括黑色素瘤、非小细胞肺癌、胃癌、尿路上皮癌等。随着临床研究的开展，单抗药物有望在更多的适应症中实现突破。Immunotherapy based on PD-1/PD-L1 has attracted much attention, and the PD-1/PD-L1 monoclonal antibodies that have been approved for marketing include Pembrolizumab of Merck, Nivolumab of Bristol-Myers Squibb, Avelumab of Merck, AstraZeneca Durvalumab, Roche's Atezolizumab, etc. The above monoclonal antibodies have shown significant efficacy in the treatment of various tumor types, and the approved indications include melanoma, non-small cell lung cancer, gastric cancer, urothelial cancer, etc. With the development of clinical research, monoclonal antibodies are expected to achieve breakthroughs in more indications.
虽然单抗药物在临床治疗中显示出优势，但也存在明显的缺陷如制备和纯化困难、生产成本高昂；易被蛋白酶分解，半衰期短；不能口服，只能注射给药；单抗的免疫原性导致严重的毒副作用。相比于生物大分子药物，小分子化合物经化学修饰后药物代谢动力学性质可控，另外在生产工艺、给药方式等方面也具有更大的探索与优化空间。因此，开发靶向PD-1/PD-L1蛋白/蛋白相互作用的小分子抑制剂是实现免疫治疗的可行性选择。Although monoclonal antibody drugs show advantages in clinical treatment, they also have obvious defects such as difficult preparation and purification, high production cost; easy to be decomposed by protease, short half-life; can not be taken orally, only by injection; the immunogen of monoclonal antibody Sex leads to serious toxic side effects. Compared with biological macromolecular drugs, the pharmacokinetic properties of small molecular compounds after chemical modification are controllable, and they also have greater space for exploration and optimization in terms of production technology and administration methods. Therefore, the development of small-molecule inhibitors targeting PD-1/PD-L1 protein/protein interactions is a feasible option for realizing immunotherapy.
目前小分子PD-1/PD-L1抑制剂研发处于早期阶段，因此，迫切需要开发具有新颖化学结构的小分子PD-1/PD-L1抑制剂。At present, the development of small molecule PD-1/PD-L1 inhibitors is at an early stage, therefore, there is an urgent need to develop small molecule PD-1/PD-L1 inhibitors with novel chemical structures.
发明内容：Invention content:
本发明人在参考文献基础上，设计并合成了一系列三唑并吡啶类化合物。活性研究结果表明，该类化合物可以显著抑制PD-1/PD-L1蛋白/蛋白相互作用。Based on the references, the inventors designed and synthesized a series of triazolopyridine compounds. The results of the activity study showed that these compounds can significantly inhibit the PD-1/PD-L1 protein/protein interaction.
本发明涉及通式Ⅰ的三唑并吡啶类化合物及其立体异构体以及药学上可接受的盐，The present invention relates to triazolopyridine compounds of general formula I and their stereoisomers and pharmaceutically acceptable salts,
其中，in,
R1选自无取代或被R5取代的苯基、
R5独立地选自卤素、(C1-C4)烷基、(C1-C4)烷氧基、氰基、羟基、羧基、氨基；R 5 is independently selected from halogen, (C 1 -C 4 )alkyl, (C 1 -C 4 )alkoxy, cyano, hydroxyl, carboxyl, amino;
R2选自卤素、氰基、(C1-C4)烷基；R 2 is selected from halogen, cyano, (C 1 -C 4 )alkyl;
R3、R4独立地选自氢、(C1-C4)烷基、(C3-C8)环烷基、羟基(C1-C4)烷基、氨基(C1-C4)烷基、氨基甲酰基(C1-C4)烷基、氨基磺酰基(C1-C4)烷基、甲磺酰胺基(C1-C4)烷基、羧基(C1-C4)烷基、(C1-C4)烷氧甲酰基(C1-C4)烷基；所述(C1-C4)烷基、(C3-C8)环烷基、羟基(C1-C4)烷基、氨基(C1-C4)烷基、氨基甲酰基(C1-C4)烷基、氨基磺酰基(C1-C4)烷基、甲磺酰胺基(C1-C4)烷基、羧基(C1-C4)烷基、(C1-C4)烷氧甲酰基(C1-C4)烷基可任选被1-3个R6取代；R 3 , R 4 are independently selected from hydrogen, (C 1 -C 4 )alkyl, (C 3 -C 8 )cycloalkyl, hydroxy (C 1 -C 4 )alkyl, amino (C 1 -C 4 ) ) alkyl, carbamoyl (C 1 -C 4 ) alkyl, aminosulfonyl (C 1 -C 4 ) alkyl, methanesulfonamido (C 1 -C 4 ) alkyl, carboxyl (C 1 -C 4 ) alkyl 4 ) alkyl, (C 1 -C 4 ) alkoxycarbonyl (C 1 -C 4 ) alkyl; the (C 1 -C 4 ) alkyl, (C 3 -C 8 ) cycloalkyl, hydroxyl (C 1 -C 4 )alkyl, amino(C 1 -C 4 )alkyl, carbamoyl(C 1 -C 4 )alkyl, aminosulfonyl(C 1 -C 4 )alkyl, methanesulfonamide (C 1 -C 4 ) alkyl group, carboxy (C 1 -C 4 ) alkyl group, (C 1 -C 4 ) alkoxycarbonyl (C 1 -C 4 ) alkyl group can be optionally replaced by 1-3 R 6 substituted;
或者R3、R4和与它们相连的氮原子一起形成一个3-7元的含氮杂环；所述含氮杂环可任选被1-3个R7取代；Or R 3 , R 4 and the nitrogen atom to which they are attached together form a 3-7 membered nitrogen-containing heterocycle; the nitrogen-containing heterocycle may be optionally substituted by 1-3 R 7 ;
R6独立地选自氢、卤素、羟基、羧基、氨基、(C1-C4)烷基、羟基(C1-C4)烷基、(C1-C4)烷氧基(C1-C4)烷基、羧基(C1-C4)烷基、(C1-C4)烷氧甲酰基(C1-C4)烷基、(C1-C4)酰基；R 6 is independently selected from hydrogen, halogen, hydroxy, carboxyl, amino, (C 1 -C 4 )alkyl, hydroxy(C 1 -C 4 )alkyl, (C 1 -C 4 )alkoxy (C 1 ) -C 4 ) alkyl, carboxy(C 1 -C 4 ) alkyl, (C 1 -C 4 )alkoxycarbonyl(C 1 -C 4 ) alkyl, (C 1 -C 4 ) acyl;
R7独立地选自氢、羟基、羧基、氨基、氨甲酰基、(C1-C4)烷基、(C1-C4)烷氧基、(C1-C4)烷氧甲酰基、羟基(C1-C4)烷基。R 7 is independently selected from hydrogen, hydroxyl, carboxyl, amino, carbamoyl, (C 1 -C 4 )alkyl, (C 1 -C 4 )alkoxy, (C 1 -C 4 )alkoxyformyl , hydroxy (C 1 -C 4 ) alkyl.
本发明优选涉及通式Ⅰ的三唑并吡啶类化合物及其立体异构体以及药学上可接受的盐，The present invention preferably relates to triazolopyridine compounds of general formula I and their stereoisomers and pharmaceutically acceptable salts,
其中，in,
R1选自无取代或被卤素、甲基、甲氧基取代的苯基、
R2选自卤素、氰基、(C1-C4)烷基；R 2 is selected from halogen, cyano, (C 1 -C 4 )alkyl;
R3、R4独立地选自氢、(C1-C4)烷基、(C3-C8)环烷基、羟基(C1-C4)烷基、氨基(C1-C4)烷基、氨基甲酰基(C1-C4)烷基、氨基磺酰基(C1-C4)烷基、甲磺酰胺基(C1-C4)烷基、羧基(C1-C4)烷基、(C1-C4)烷氧甲酰基(C1-C4)烷基；所述(C1-C4)烷基、(C3-C8)环烷基、羟基(C1-C4)烷基、氨基(C1-C4)烷基、氨基甲酰基(C1-C4)烷基、氨基磺酰基(C1-C4)烷基、甲磺酰胺基(C1-C4)烷基、羧基(C1-C4)烷基、(C1-C4)烷氧甲酰基(C1-C4)烷基可任选被1-3个R6取代；R 3 , R 4 are independently selected from hydrogen, (C 1 -C 4 )alkyl, (C 3 -C 8 )cycloalkyl, hydroxy (C 1 -C 4 )alkyl, amino (C 1 -C 4 ) ) alkyl, carbamoyl (C 1 -C 4 ) alkyl, aminosulfonyl (C 1 -C 4 ) alkyl, methanesulfonamido (C 1 -C 4 ) alkyl, carboxyl (C 1 -C 4 ) alkyl 4 ) alkyl, (C 1 -C 4 ) alkoxycarbonyl (C 1 -C 4 ) alkyl; the (C 1 -C 4 ) alkyl, (C 3 -C 8 ) cycloalkyl, hydroxyl (C 1 -C 4 )alkyl, amino(C 1 -C 4 )alkyl, carbamoyl(C 1 -C 4 )alkyl, aminosulfonyl(C 1 -C 4 )alkyl, methanesulfonamide (C 1 -C 4 ) alkyl group, carboxy (C 1 -C 4 ) alkyl group, (C 1 -C 4 ) alkoxycarbonyl (C 1 -C 4 ) alkyl group can be optionally replaced by 1-3 R 6 substituted;
或者R3、R4和与它们相连的氮原子一起形成一个3-7元的含氮杂环；所述含氮杂环可任选被1-3个R7取代；Or R 3 , R 4 and the nitrogen atom to which they are attached together form a 3-7 membered nitrogen-containing heterocycle; the nitrogen-containing heterocycle may be optionally substituted by 1-3 R 7 ;
R6独立地选自氢、卤素、羟基、羧基、氨基、(C1-C4)烷基、羟基(C1-C4)烷基、(C1-C4)烷氧基(C1-C4)烷基、羧基(C1-C4)烷基、(C1-C4)烷氧甲酰基(C1-C4)烷基、(C1-C4)酰基；R 6 is independently selected from hydrogen, halogen, hydroxy, carboxyl, amino, (C 1 -C 4 )alkyl, hydroxy(C 1 -C 4 )alkyl, (C 1 -C 4 )alkoxy (C 1 ) -C 4 ) alkyl, carboxy(C 1 -C 4 ) alkyl, (C 1 -C 4 )alkoxycarbonyl(C 1 -C 4 ) alkyl, (C 1 -C 4 ) acyl;
R7独立地选自氢、羟基、羧基、氨基、氨甲酰基、(C1-C4)烷基、(C1-C4)烷氧基、(C1-C4)烷氧甲酰基、羟基(C1-C4)烷基。R 7 is independently selected from hydrogen, hydroxyl, carboxyl, amino, carbamoyl, (C 1 -C 4 )alkyl, (C 1 -C 4 )alkoxy, (C 1 -C 4 )alkoxyformyl , hydroxy (C 1 -C 4 ) alkyl.
本发明更加优选涉及通式Ⅰ的三唑并吡啶类化合物及其立体异构体以及药学上可接受的盐，其中，The present invention more preferably relates to triazolopyridine compounds of general formula I and their stereoisomers and pharmaceutically acceptable salts, wherein,
R1选自无取代或被1-3个卤素、甲基、甲氧基取代的苯基、
R2选自卤素、氰基、甲基；R 2 is selected from halogen, cyano, methyl;
R3、R4独立地选自氢、(C1-C4)烷基、(C3-C8)环烷基、羟基(C1-C4)烷基、氨基(C1-C4)烷基、氨基甲酰基(C1-C4)烷基、氨基磺酰基(C1-C4)烷基、甲磺酰胺基(C1-C4)烷基、羧基(C1-C4)烷基、(C1-C4)烷氧甲酰基(C1-C4)烷基；所述(C1-C4)烷基、(C3-C8)环烷基、羟基(C1-C4)烷基、氨基(C1-C4)烷基、氨基甲酰基(C1-C4)烷基、氨基磺酰基(C1-C4)烷基、甲磺酰胺基(C1-C4)烷基、羧基(C1-C4)烷基、(C1-C4)烷氧甲酰基(C1-C4)烷基可任选被1-3个R6取代；R 3 , R 4 are independently selected from hydrogen, (C 1 -C 4 )alkyl, (C 3 -C 8 )cycloalkyl, hydroxy (C 1 -C 4 )alkyl, amino (C 1 -C 4 ) ) alkyl, carbamoyl (C 1 -C 4 ) alkyl, aminosulfonyl (C 1 -C 4 ) alkyl, methanesulfonamido (C 1 -C 4 ) alkyl, carboxyl (C 1 -C 4 ) alkyl 4 ) alkyl, (C 1 -C 4 ) alkoxycarbonyl (C 1 -C 4 ) alkyl; the (C 1 -C 4 ) alkyl, (C 3 -C 8 ) cycloalkyl, hydroxyl (C 1 -C 4 )alkyl, amino(C 1 -C 4 )alkyl, carbamoyl(C 1 -C 4 )alkyl, aminosulfonyl(C 1 -C 4 )alkyl, methanesulfonamide (C 1 -C 4 ) alkyl group, carboxy (C 1 -C 4 ) alkyl group, (C 1 -C 4 ) alkoxycarbonyl (C 1 -C 4 ) alkyl group can be optionally replaced by 1-3 R 6 substituted;
或者R3、R4和与它们相连的氮原子一起形成一个5-6元的含氮杂环；所述含氮杂环可任选被1-3个R7取代；Or R 3 , R 4 and the nitrogen atom to which they are attached together form a 5-6 membered nitrogen-containing heterocyclic ring; the nitrogen-containing heterocyclic ring can be optionally substituted by 1-3 R 7 ;
R6独立地选自氢、卤素、羟基、羧基、氨基、(C1-C4)烷基、羟基(C1-C4)烷基、(C1-C4)烷氧基(C1-C4)烷基、羧基(C1-C4)烷基、(C1-C4)烷氧甲酰基(C1-C4)烷基、(C1-C4)酰基；R 6 is independently selected from hydrogen, halogen, hydroxy, carboxyl, amino, (C 1 -C 4 )alkyl, hydroxy(C 1 -C 4 )alkyl, (C 1 -C 4 )alkoxy (C 1 ) -C 4 ) alkyl, carboxy(C 1 -C 4 ) alkyl, (C 1 -C 4 )alkoxycarbonyl(C 1 -C 4 ) alkyl, (C 1 -C 4 ) acyl;
R7独立地选自氢、羟基、羧基、氨基、氨甲酰基、(C1-C4)烷基、(C1-C4)烷氧基、(C1-C4)烷氧甲酰基、羟基(C1-C4)烷基。R 7 is independently selected from hydrogen, hydroxyl, carboxyl, amino, carbamoyl, (C 1 -C 4 )alkyl, (C 1 -C 4 )alkoxy, (C 1 -C 4 )alkoxyformyl , hydroxy (C 1 -C 4 ) alkyl.
本发明特别优选涉及通式Ⅰ的三唑并吡啶类化合物及其立体异构体以及药学上可接受的盐，其中，The present invention particularly preferably relates to triazolopyridine compounds of general formula I and their stereoisomers and pharmaceutically acceptable salts, wherein,
R1选自无取代或被1-3个卤素、甲基、甲氧基取代的苯基、
R2选自氯、氰基、甲基；R 2 is selected from chlorine, cyano, methyl;
本发明通式Ⅰ的三唑并吡啶类化合物及其立体异构体以及药学上可接受的盐优选自以下化合物，但这些化合物并不意味着对本发明的任何限制：The triazolopyridine compounds of the general formula I and their stereoisomers and pharmaceutically acceptable salts of the present invention are preferably selected from the following compounds, but these compounds do not mean any limitation to the present invention:
此外，本发明还包括本发明化合物的前药。本发明化合物的前药是通式Ⅰ的衍生物，它们自身可能具有较弱的活性甚至没有活性，但是在给药后，在生理条件下(例如通过代谢、溶剂分解或另外的方式)被转化成相应的生物活性形式。In addition, the present invention also includes prodrugs of the compounds of the present invention. Prodrugs of the compounds of the present invention are derivatives of general formula I, which themselves may have weak or even no activity, but are converted under physiological conditions (eg by metabolism, solvolysis or otherwise) after administration into the corresponding biologically active form.
以上所述的通式Ⅰ的三唑并吡啶类化合物及其立体异构体以及药学上可接受的盐，所述的药学上可接受的盐包括与无机酸、有机酸、碱金属离子形成的盐；所述的无机酸选自：盐酸、氢溴酸、氢氟酸、硫酸、磷酸；所述的有机酸选自：琥珀酸、富马酸、马来酸、乳酸、苹果酸、酒石酸、柠檬酸、甲磺酸、乙磺酸或对甲苯磺酸；所属的碱金属离子选自锂离子、钠离子或钾离子。The above-mentioned triazolopyridine compounds of the general formula I and their stereoisomers and pharmaceutically acceptable salts, the pharmaceutically acceptable salts include those formed with inorganic acids, organic acids and alkali metal ions. Salt; described inorganic acid is selected from: hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, phosphoric acid; described organic acid is selected from: succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, Citric acid, methanesulfonic acid, ethanesulfonic acid or p-toluenesulfonic acid; the associated alkali metal ions are selected from lithium, sodium or potassium ions.
本发明中“卤素”是指氟、氯、溴或碘代；“烷基”是指直链或支链的烷基；“含氮杂环”是指含有氮原子的单环或多环的环状体系，环状体系是非芳香性或芳香性的。In the present invention, "halogen" refers to fluorine, chlorine, bromine or iodine; "alkyl" refers to a straight-chain or branched alkyl group; "nitrogen-containing heterocycle" refers to a monocyclic or polycyclic ring containing a nitrogen atom. Ring systems, which are non-aromatic or aromatic.
本发明可以含有上式Ⅰ的三唑并吡啶类化合物及其立体异构体以及药学上可接受的盐作为活性成分，与药学上可接受的载体或赋形剂混合制备成组合物。所述载体或赋形剂包括本领域公知的稀释剂、粘合剂、润湿剂、崩解剂、润滑剂、助流剂等。稀释剂包括但不限于淀粉、糊精、蔗糖、葡萄糖、乳糖、甘露醇、山梨醇、木糖醇、磷酸氢钙等；润湿剂包括水、乙醇、异丙醇等；粘合剂包括但不限于淀粉浆、糊精、糖浆、蜂蜜、葡萄糖溶液、阿拉伯胶浆、明胶浆、羧甲基纤维素钠、羟丙基甲基纤维素、乙基纤维素、聚乙二醇等；崩解剂包括但不限于干淀粉、微晶纤维素、低取代羟丙基纤维素、交联聚乙烯吡咯烷酮、交联羧甲基纤维素钠、羧甲基淀粉钠、十二烷基磺酸钠等；润滑剂和助流剂包括但不限于滑石粉、二氧化硅、聚乙二醇等。The present invention can contain the triazolopyridine compounds of the above formula I and their stereoisomers and pharmaceutically acceptable salts as active ingredients, mixed with pharmaceutically acceptable carriers or excipients to prepare a composition. The carriers or excipients include diluents, binders, wetting agents, disintegrants, lubricants, glidants and the like known in the art. Diluents include but are not limited to starch, dextrin, sucrose, glucose, lactose, mannitol, sorbitol, xylitol, calcium hydrogen phosphate, etc.; wetting agents include water, ethanol, isopropanol, etc.; binders include but Not limited to starch slurry, dextrin, syrup, honey, glucose solution, acacia mucilage, gelatin slurry, sodium carboxymethyl cellulose, hydroxypropyl methyl cellulose, ethyl cellulose, polyethylene glycol, etc.; disintegration Agents include but are not limited to dry starch, microcrystalline cellulose, low-substituted hydroxypropyl cellulose, cross-linked polyvinyl pyrrolidone, cross-linked sodium carboxymethyl cellulose, sodium carboxymethyl starch, sodium lauryl sulfonate, etc. ; Lubricants and glidants include, but are not limited to, talc, silicon dioxide, polyethylene glycol, and the like.
本发明的药用组合物可配制成若干种剂型，所述剂型包括但不限于注射剂、片剂、胶囊剂等。The pharmaceutical composition of the present invention can be formulated into several dosage forms, including but not limited to injections, tablets, capsules, and the like.
本发明的三唑并吡啶类化合物及其立体异构体以及药学上可接受的盐可以与其他活性成分组合使用，从而达到更优的治疗效果。The triazolopyridine compounds and their stereoisomers and pharmaceutically acceptable salts of the present invention can be used in combination with other active ingredients to achieve better therapeutic effects.
本发明还提供了通式Ⅰ的三唑并吡啶类化合物及其立体异构体以及药学上可接受的盐在制备预防和/或治疗与PD-1/PD-L1信号通路有关疾病的药物中的应用。所述的与PD-1/PD-L1信号通路有关的疾病选自癌症、感染性疾病、自身免疫性疾病。所述的癌症选自肺癌、皮肤癌、血液肿瘤、胶质瘤、消化系统肿瘤、乳腺癌、淋巴瘤、神经系统肿瘤、黑色素瘤；所述的感染性疾病选自细菌感染、病毒感染；所述的自身免疫性疾病选自器官特异性自身免疫病、系统性自身免疫病。其中，所述器官特异性自身免疫病包括慢性淋巴细胞性甲状腺炎、甲状腺功能亢进、胰岛素依赖型糖尿病、溃疡性结肠炎、急性特发性多神经炎，所述的系统性自身免疫病包括类风湿性关节炎、系统性红斑狼疮、系统性血管炎、自身免疫性溶血性贫血。The present invention also provides triazolopyridine compounds of general formula I, stereoisomers thereof and pharmaceutically acceptable salts in the preparation of medicaments for preventing and/or treating diseases related to PD-1/PD-L1 signaling pathway Applications. The diseases related to the PD-1/PD-L1 signaling pathway are selected from cancer, infectious diseases, and autoimmune diseases. The cancer is selected from lung cancer, skin cancer, hematological tumor, glioma, digestive system tumor, breast cancer, lymphoma, nervous system tumor, and melanoma; the infectious disease is selected from bacterial infection and viral infection; Said autoimmune disease is selected from organ-specific autoimmune disease and systemic autoimmune disease. Wherein, the organ-specific autoimmune diseases include chronic lymphocytic thyroiditis, hyperthyroidism, insulin-dependent diabetes mellitus, ulcerative colitis, acute idiopathic polyneuritis, and the systemic autoimmune diseases include Rheumatoid arthritis, systemic lupus erythematosus, systemic vasculitis, autoimmune hemolytic anemia.
本发明的积极进步效果在于：本发明的三唑并吡啶类化合物具有新颖的化学结构，并在体外研究中对PD-1/PD-L1蛋白/蛋白相互作用具有很高的抑制活性，可用于癌症等多种疾病的治疗和预防。The positive improvement effect of the present invention is that: the triazolopyridine compound of the present invention has a novel chemical structure, and has a high inhibitory activity on PD-1/PD-L1 protein/protein interaction in in vitro studies, and can be used for Treatment and prevention of various diseases such as cancer.
下文中提供的实施例和制备例进一步阐明和举例说明本发明化合物及其制备方法。应当理解，下述实例和制备例的范围并不以任何方式限制本发明的范围。The examples and preparations provided hereinafter further illustrate and illustrate the compounds of the present invention and methods for their preparation. It should be understood that the scope of the following examples and preparations do not limit the scope of the invention in any way.
下面的合成路线概括并描述了本发明的式Ⅰ衍生物的制备，所有的原料都是通过这些流程中描述的方式、通过有机化学领域普通技术人员熟知的方法制备的或者可商购。本发明的全部最终衍生物都是通过这些流程中描述的方法或通过与其类似的方法制备的，这些方法是有机化学领域普通技术人员熟知的。这些流程中应用的全部可变因数如权利要求中的定义。The following synthetic schemes outline and describe the preparation of the derivatives of formula I of the present invention, all starting materials were prepared by the means described in these schemes, by methods well known to those of ordinary skill in the art of organic chemistry, or are commercially available. All final derivatives of the present invention are prepared by methods described in these schemes or by methods analogous thereto, which are well known to those of ordinary skill in the art of organic chemistry. All variable factors applied in these procedures are as defined in the claims.
(a)以取代的苯胺衍生物1和苯、取代苯或杂芳环硼酸或硼酸酯为原料，通过Suzuki偶联反应得到中间体2；(a) Using substituted aniline derivative 1 and benzene, substituted benzene or heteroaromatic boronic acid or boronic acid ester as raw materials, obtain intermediate 2 through Suzuki coupling reaction;
(b)以5-溴-2-氯烟酸甲酯3为原料，与水合肼反应得到中间体4；(b) take 5-bromo-2-chloronicotinic acid methyl ester 3 as raw material, react with hydrazine hydrate to obtain intermediate 4;
(c)以中间体4为原料，与原甲酸三甲酯反应得到三唑并吡啶中间体5；(c) take intermediate 4 as raw material, react with trimethyl orthoformate to obtain triazolopyridine intermediate 5;
(d)以中间体5为原料，在碱性条件下水解反应得到羧酸中间体6；(d) using intermediate 5 as a raw material, hydrolysis reaction under alkaline conditions obtains carboxylic acid intermediate 6;
(e)以中间体6和中间体2为原料，在缩合剂条件下经过酰胺化反应得到中间体7；或由中间体6制备酰氯再与中间体2反应得到中间体7；(e) take intermediate 6 and intermediate 2 as raw materials, and obtain intermediate 7 through amidation reaction under the condition of a condensing agent; or prepare acid chloride from intermediate 6 and then react with intermediate 2 to obtain intermediate 7;
(f)以中间体7为原料，与乙烯基硼酸频哪醇酯通过Suzuki偶联反应得到中间体8；(f) take intermediate 7 as raw material, and obtain intermediate 8 through Suzuki coupling reaction with vinylboronic acid pinacol ester;
(g)以中间体8为原料，在高碘酸钠等氧化条件下制备得到醛类化合物中间体9；(g) take intermediate 8 as raw material, prepare aldehyde compound intermediate 9 under oxidation conditions such as sodium periodate;
(h)以中间体9为原料，与胺类化合物HNR3R4缩合及氰基硼氢化钠或三乙酰氧基硼氢化钠作用下还原得到通式Ⅰ的目标化合物。(h) Using intermediate 9 as raw material, condensing with amine compound HNR 3 R 4 and reducing under the action of sodium cyanoborohydride or sodium triacetoxyborohydride to obtain the target compound of general formula I.
所述的R1、R2、R3、R4的定义如权利要求所述。在中间体1中，当R2为(C1-C4)烷基时，X为氯、溴或碘；当R2为氰基时，X为溴或碘；当R2为氯时，X为溴或碘；当R2为溴时，X为碘。The definitions of R 1 , R 2 , R 3 and R 4 are as described in the claims. In Intermediate 1, when R 2 is (C 1 -C 4 )alkyl, X is chlorine, bromine or iodine; when R 2 is cyano, X is bromine or iodine; when R 2 is chlorine, X is bromine or iodine; when R 2 is bromine, X is iodine.
本发明的具有通式Ⅰ的三唑并吡啶类化合物均可按照上述反应路线制备得到。The triazolopyridine compounds having the general formula I of the present invention can be prepared according to the above reaction scheme.
具体实施方式：Detailed ways:
在以下的实施例中，描绘了制备部分所述化合物的方法。应了解，以下方法及所属领域的普通技术人员已知的其他方法均可适用于本发明所述的所有化合物的制备。实施例旨在阐述而不是限制本发明的范围。In the following examples, methods for preparing some of the compounds are depicted. It will be appreciated that the following methods, as well as other methods known to those of ordinary skill in the art, are applicable to the preparation of all compounds described herein. The examples are intended to illustrate rather than limit the scope of the invention.
实施例1：6-(((2-羟乙基)氨基)甲基)-N-(2-甲基-[1,1'-联苯]-3-基)-[1,2,4]三唑并[4,3-a]吡啶-8-甲酰胺(I-1)Example 1: 6-(((2-hydroxyethyl)amino)methyl)-N-(2-methyl-[1,1'-biphenyl]-3-yl)-[1,2,4 ]Triazolo[4,3-a]pyridine-8-carboxamide (I-1)
步骤1：2-甲基-[1,1'-联苯]-3-胺Step 1: 2-Methyl-[1,1'-biphenyl]-3-amine
室温下，将3-溴-2-甲基苯胺(15g，0.081mol)、苯硼酸(12.2g，0.1mol)、醋酸钯(1.82g，8.1mmol)、碳酸钾(24.9g，0.18mol)加入至乙醇/水的混合溶液中(体积比1：1，100mL)，于N2保护下搅拌反应5小时。反映完毕后，抽滤，蒸干滤液，柱层析分离得浅黄色固体12.6g，收率85.2％。At room temperature, 3-bromo-2-methylaniline (15g, 0.081mol), phenylboronic acid (12.2g, 0.1mol), palladium acetate (1.82g, 8.1mmol), potassium carbonate (24.9g, 0.18mol) were added into a mixed solution of ethanol/water (volume ratio 1:1, 100 mL), and stirred for 5 hours under the protection of N 2 . After completion of the reaction, suction filtration, the filtrate was evaporated to dryness, and 12.6 g of a light yellow solid was obtained by column chromatography with a yield of 85.2%.
步骤2：5-溴-2-肼基烟酸甲酯Step 2: Methyl 5-bromo-2-hydrazinonicotinate
室温下，将5-溴-2-氯烟酸甲酯(13.55g，0.054mol)溶于1，4-二氧六环(150mL)中，加入水合肼(6g，0.096mol)，60℃下反应3小时。反应完毕后，向溶液中加入50mL水，搅拌0.5小时，抽滤，干燥，得黄色固体10.2g，收率为77.4％。At room temperature, methyl 5-bromo-2-chloronicotinate (13.55 g, 0.054 mol) was dissolved in 1,4-dioxane (150 mL), hydrazine hydrate (6 g, 0.096 mol) was added, and the mixture was heated at 60°C. The reaction was carried out for 3 hours. After the reaction was completed, 50 mL of water was added to the solution, stirred for 0.5 hour, suction filtered, and dried to obtain 10.2 g of a yellow solid with a yield of 77.4%.
步骤3：6-溴-[1,2,4]三唑并[4,3-a]吡啶-8-甲酸甲酯Step 3: Methyl 6-bromo-[1,2,4]triazolo[4,3-a]pyridine-8-carboxylate
将5-溴-2-肼基烟酸甲酯(5g，0.02mol)溶于原甲酸三甲酯(25mL)中，于100℃反应12小时。将反应液冷却至室温，搅拌0.5小时，抽滤，水洗滤饼，干燥，得白色固体4.6g，收率为88.5％。Methyl 5-bromo-2-hydrazinonicotinate (5 g, 0.02 mol) was dissolved in trimethyl orthoformate (25 mL) and reacted at 100° C. for 12 hours. The reaction solution was cooled to room temperature, stirred for 0.5 hours, suction filtered, the filter cake was washed with water, and dried to obtain 4.6 g of a white solid with a yield of 88.5%.
步骤4：6-溴-[1,2,4]三唑并[4,3-a]吡啶-8-甲酸Step 4: 6-Bromo-[1,2,4]triazolo[4,3-a]pyridine-8-carboxylic acid
室温下，将6-溴-[1,2,4]三唑并[4,3-a]吡啶-8-甲酸甲酯(1g，0.004mol)溶于甲醇(5mL)中，加入1mol/L的NaOH水溶液(10mL)，升温至回流反应0.5小时。将反应液冷却至室温，蒸除甲醇，向体系中加入少量水，用稀盐酸调节pH至5-6，搅拌20分钟，抽滤得白色固体0.67g，收率为70.9％。At room temperature, methyl 6-bromo-[1,2,4]triazolo[4,3-a]pyridine-8-carboxylate (1 g, 0.004 mol) was dissolved in methanol (5 mL), and 1 mol/L was added. NaOH aqueous solution (10 mL) was heated to reflux for 0.5 hours. The reaction solution was cooled to room temperature, methanol was evaporated, a small amount of water was added to the system, the pH was adjusted to 5-6 with dilute hydrochloric acid, stirred for 20 minutes, and suction filtered to obtain 0.67 g of a white solid with a yield of 70.9%.
步骤5：6-溴-N-(2-甲基-[1,1'-联苯]-3-基)-[1,2,4]三唑并[4,3-a]吡啶-8-甲酰胺Step 5: 6-Bromo-N-(2-methyl-[1,1'-biphenyl]-3-yl)-[1,2,4]triazolo[4,3-a]pyridine-8 -Formamide
室温下，将6-溴-[1,2,4]三唑并[4,3-a]吡啶-8-甲酸(2.18g，9mmol)、2-甲基-[1,1'-联苯]-3-胺(1.65g，9.5mmol)、HATU(5.13g，13.5mmol)、N，N-二异丙基乙胺(2.32g，18mmol)溶于N,N-二甲基甲酰胺(15mL)中，搅拌反应15小时。反应完毕后，将反应液倒入大量水中，搅拌20分钟，抽滤，水洗滤饼，干燥，柱层析分离得黄色固体2.97g，收率为81.3％。At room temperature, 6-bromo-[1,2,4]triazolo[4,3-a]pyridine-8-carboxylic acid (2.18 g, 9 mmol), 2-methyl-[1,1'-biphenyl ]-3-amine (1.65 g, 9.5 mmol), HATU (5.13 g, 13.5 mmol), N,N-diisopropylethylamine (2.32 g, 18 mmol) were dissolved in N,N-dimethylformamide ( 15 mL), the reaction was stirred for 15 hours. After the reaction was completed, the reaction solution was poured into a large amount of water, stirred for 20 minutes, suction filtered, the filter cake was washed with water, dried, and separated by column chromatography to obtain 2.97 g of a yellow solid with a yield of 81.3%.
步骤6：N-(2-甲基-[1,1'-联苯]-3-基)-6-乙烯基-[1,2,4]三唑并[4,3-a]吡啶-8-甲酰胺Step 6: N-(2-Methyl-[1,1'-biphenyl]-3-yl)-6-vinyl-[1,2,4]triazolo[4,3-a]pyridine- 8-Carboxamide
室温下，将6-溴-N-(2-甲基-[1,1'-联苯]-3-基)-[1,2,4]三唑并[4,3-a]吡啶-8-甲酰胺(2.3g，5.7mmol)、乙烯基硼酸频哪醇酯(0.96g，6.2mmol)、[1，1’-双(二苯基膦基)二茂铁]二氯化钯(0.42g，0.57mmol)、碳酸铯(3.4g，10.3mmol)溶于1，4-二氧六环/水(体积比3：1，10mL)的混合溶液中，N2保护下90℃反应2小时。反应液冷却至室温，用乙酸乙酯萃取，蒸干有机层，柱层析分离得白色固体1.21g，收率为60％。At room temperature, 6-bromo-N-(2-methyl-[1,1'-biphenyl]-3-yl)-[1,2,4]triazolo[4,3-a]pyridine- 8-Carboxamide (2.3 g, 5.7 mmol), vinylboronic acid pinacol ester (0.96 g, 6.2 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride ( 0.42g, 0.57mmol) and cesium carbonate (3.4g, 10.3mmol) were dissolved in a mixed solution of 1,4-dioxane/water (volume ratio 3:1, 10mL), and reacted at 90°C under the protection of N2 for 2 Hour. The reaction solution was cooled to room temperature, extracted with ethyl acetate, the organic layer was evaporated to dryness, and 1.21 g of a white solid was obtained by column chromatography with a yield of 60%.
步骤7：6-甲酰基-N-(2-甲基-[1,1'-联苯]-3-基)-[1,2,4]三唑并[4,3-a]吡啶-8-甲酰胺Step 7: 6-Formyl-N-(2-methyl-[1,1'-biphenyl]-3-yl)-[1,2,4]triazolo[4,3-a]pyridine- 8-Carboxamide
室温下，将N-(2-甲基-[1,1'-联苯]-3-基)-6-乙烯基-[1,2,4]三唑并[4,3-a]吡啶-8-甲酰胺(0.65g，1.8mmol)溶于1，4-二氧六环/水(体积比5：1，10mL)的混合溶液中，快速加入2％的OsO4水溶液(3.5mL)，搅拌5分钟后加入高碘酸钠(1.54g，7.2mmol)，室温搅拌反应9小时。抽滤，得浅黄色固体0.41g，收率为64.2％。At room temperature, N-(2-methyl-[1,1'-biphenyl]-3-yl)-6-vinyl-[1,2,4]triazolo[4,3-a]pyridine -8-Carboxamide (0.65g, 1.8mmol) was dissolved in a mixed solution of 1,4-dioxane/water (volume ratio 5:1, 10mL), and a 2% OsO4 aqueous solution (3.5mL) was quickly added After stirring for 5 minutes, sodium periodate (1.54 g, 7.2 mmol) was added, and the reaction was stirred at room temperature for 9 hours. Suction filtration to obtain 0.41 g of a light yellow solid with a yield of 64.2%.
步骤8：6-(((2-羟乙基)氨基)甲基)-N-(2-甲基-[1,1'-联苯]-3-基)-[1,2,4]三唑并[4,3-a]吡啶-8-甲酰胺(实施例1)Step 8: 6-(((2-Hydroxyethyl)amino)methyl)-N-(2-methyl-[1,1'-biphenyl]-3-yl)-[1,2,4] Triazolo[4,3-a]pyridine-8-carboxamide (Example 1)
室温下，将6-甲酰基-N-(2-甲基-[1,1'-联苯]-3-基)-[1,2,4]三唑并[4,3-a]吡啶-8-甲酰胺(0.08g，0.22mmol)溶于DMF中，加入乙醇胺(0.067g，1.1mmol)、冰醋酸(0.02g，0.33mmol)，搅拌反应2小时。向体系中加入氰基硼氢化钠(0.07g，1.1mmol)，25℃搅拌反应12小时。反应液中加水，用二氯甲烷萃取，蒸干溶剂，柱层析分离得白色固体0.032g，收率为35.6％。At room temperature, 6-formyl-N-(2-methyl-[1,1'-biphenyl]-3-yl)-[1,2,4]triazolo[4,3-a]pyridine -8-Carboxamide (0.08 g, 0.22 mmol) was dissolved in DMF, ethanolamine (0.067 g, 1.1 mmol) and glacial acetic acid (0.02 g, 0.33 mmol) were added, and the reaction was stirred for 2 hours. Sodium cyanoborohydride (0.07 g, 1.1 mmol) was added to the system, and the reaction was stirred at 25°C for 12 hours. Water was added to the reaction solution, extracted with dichloromethane, the solvent was evaporated to dryness, and 0.032 g of a white solid was obtained by column chromatography with a yield of 35.6%.
ESI-MS m/z:402.2[M+H]+；1H NMR(400MHz,DMSO-d6)δ11.41(s,1H),9.20(s,1H),8.77(s,1H),8.56(d,J＝1.3Hz,1H),8.20(d,J＝7.9Hz,1H),7.48(t,J＝7.3Hz,2H),7.43–7.29(m,4H),7.10(d,J＝7.1Hz,1H),4.63(s,1H),3.99(s,2H),3.52(d,J＝4.8Hz,2H),3.17(d,J＝4.6Hz,1H),2.67(t,J＝5.7Hz,2H),2.35(s,3H).ESI-MS m/z: 402.2 [M+H] + ; 1 H NMR (400 MHz, DMSO-d 6 ) δ 11.41 (s, 1H), 9.20 (s, 1H), 8.77 (s, 1H), 8.56 (d, J=1.3Hz, 1H), 8.20 (d, J=7.9Hz, 1H), 7.48 (t, J=7.3Hz, 2H), 7.43–7.29 (m, 4H), 7.10 (d, J= 7.1Hz, 1H), 4.63(s, 1H), 3.99(s, 2H), 3.52(d, J=4.8Hz, 2H), 3.17(d, J=4.6Hz, 1H), 2.67(t, J= 5.7Hz, 2H), 2.35(s, 3H).
根据实施例1的合成方法，以6-甲酰基-N-(2-甲基-[1,1'-联苯]-3-基)-[1,2,4]三唑并[4,3-a]吡啶-8-甲酰胺为原料，通过与不同的胺类化合物反应，再经氰基硼氢化钠或三乙酰氧基硼氢化钠还原制备得到实施例2-9的化合物。According to the synthetic method of Example 1, 6-formyl-N-(2-methyl-[1,1'-biphenyl]-3-yl)-[1,2,4]triazolo[4, Using 3-a]pyridine-8-carboxamide as a raw material, the compounds of Example 2-9 are prepared by reacting with different amine compounds and then reducing by sodium cyanoborohydride or sodium triacetoxyborohydride.
实施例2：6-(((2-羟乙基)(甲基)氨基)甲基)-N-(2-甲基-[1,1'-联苯]-3-基)-[1,2,4]三唑并[4,3-a]吡啶-8-甲酰胺(I-2)Example 2: 6-(((2-hydroxyethyl)(methyl)amino)methyl)-N-(2-methyl-[1,1'-biphenyl]-3-yl)-[1 ,2,4]Triazolo[4,3-a]pyridine-8-carboxamide (I-2)
ESI-MS m/z:416.2[M+H]+；1H NMR(600MHz,DMSO-d6)δ11.40(s,1H),9.19(s,1H),8.75(s,1H),8.48(d,J＝1.3Hz,1H),8.19(d,J＝7.9Hz,1H),7.48(t,J＝7.5Hz,2H),7.42–7.33(m,4H),7.09(d,J＝7.3Hz,1H),4.51(t,J＝5.4Hz,1H),3.72(s,2H),3.56(q,J＝6.0Hz,2H),2.54(t,J＝6.1Hz,2H),2.35(s,3H),2.24(s,3H).ESI-MS m/z: 416.2 [M+H] + ; 1 H NMR (600 MHz, DMSO-d 6 ) δ 11.40 (s, 1H), 9.19 (s, 1H), 8.75 (s, 1H), 8.48 (d, J=1.3Hz, 1H), 8.19 (d, J=7.9Hz, 1H), 7.48 (t, J=7.5Hz, 2H), 7.42–7.33 (m, 4H), 7.09 (d, J= 7.3Hz, 1H), 4.51(t, J=5.4Hz, 1H), 3.72(s, 2H), 3.56(q, J=6.0Hz, 2H), 2.54(t, J=6.1Hz, 2H), 2.35 (s,3H),2.24(s,3H).
实施例3：6-(((2,3-二羟基丙基)(甲基)氨基)甲基)-N-(2-甲基-[1,1'-联苯]-3-基)-[1,2,4]三唑并[4,3-a]吡啶-8-甲酰胺(I-3)Example 3: 6-(((2,3-Dihydroxypropyl)(methyl)amino)methyl)-N-(2-methyl-[1,1'-biphenyl]-3-yl) -[1,2,4]Triazolo[4,3-a]pyridine-8-carboxamide (I-3)
ESI-MS m/z:446.2[M+H]+；1H NMR(600MHz,DMSO-d6)δ11.40(s,1H),9.20(s,1H),8.75(s,1H),8.48(s,1H),8.20(d,J＝8.0Hz,1H),7.48(t,J＝7.5Hz,2H),7.40(t,J＝7.4Hz,1H),7.38–7.36(m,2H),7.34(d,J＝7.9Hz,1H),7.09(d,J＝7.5Hz,1H),4.54(br,2H),3.74(s,2H),3.69(dd,J＝9.9,5.0Hz,1H),3.38(d,J＝5.3Hz,1H),3.32(d,J＝5.8Hz,1H),2.54(dd,J＝12.8,4.6Hz,1H),2.40(dd,J＝12.7,7.1Hz,1H),2.35(s,3H),2.25(s,3H).ESI-MS m/z: 446.2 [M+H] + ; 1 H NMR (600 MHz, DMSO-d 6 ) δ 11.40 (s, 1H), 9.20 (s, 1H), 8.75 (s, 1H), 8.48 (s, 1H), 8.20 (d, J=8.0Hz, 1H), 7.48 (t, J=7.5Hz, 2H), 7.40 (t, J=7.4Hz, 1H), 7.38–7.36 (m, 2H) ,7.34(d,J＝7.9Hz,1H),7.09(d,J＝7.5Hz,1H),4.54(br,2H),3.74(s,2H),3.69(dd,J＝9.9,5.0Hz, 1H),3.38(d,J=5.3Hz,1H),3.32(d,J=5.8Hz,1H),2.54(dd,J=12.8,4.6Hz,1H),2.40(dd,J=12.7,7.1 Hz,1H),2.35(s,3H),2.25(s,3H).
实施例4：反-6-(((4-羟基环己基)氨基)甲基)-N-(2-甲基-[1,1'-联苯]-3-基)-[1,2,4]三唑并[4,3-a]吡啶-8-甲酰胺(I-4)Example 4: trans-6-(((4-hydroxycyclohexyl)amino)methyl)-N-(2-methyl-[1,1'-biphenyl]-3-yl)-[1,2 ,4]Triazolo[4,3-a]pyridine-8-carboxamide (I-4)
ESI-MS m/z:456.2[M+H]+；1H NMR(600MHz,DMSO-d6)δ11.40(s,1H),9.14(s,1H),8.74(s,1H),8.53(s,1H),8.20(d,J＝7.8Hz,1H),7.48(t,J＝6.8Hz,2H),7.43–7.32(m,4H),7.10(d,J＝7.1Hz,1H),4.47(s,1H),3.91(s,2H),3.43(s,1H),2.41(s,1H),2.35(s,3H),1.90(d,J＝8.5Hz,2H),1.80(d,J＝9.3Hz,2H),1.24(s,1H),1.18–1.03(m,4H).ESI-MS m/z: 456.2 [M+H] + ; 1 H NMR (600 MHz, DMSO-d 6 ) δ 11.40 (s, 1H), 9.14 (s, 1H), 8.74 (s, 1H), 8.53 (s, 1H), 8.20 (d, J=7.8Hz, 1H), 7.48 (t, J=6.8Hz, 2H), 7.43–7.32 (m, 4H), 7.10 (d, J=7.1Hz, 1H) ,4.47(s,1H),3.91(s,2H),3.43(s,1H),2.41(s,1H),2.35(s,3H),1.90(d,J＝8.5Hz,2H),1.80( d, J=9.3Hz, 2H), 1.24(s, 1H), 1.18–1.03(m, 4H).
实施例5：N-(2-甲基-[1,1'-联苯]-3-基)-6-(((2-氨磺酰基乙基)氨基)甲基)-[1,2,4]三唑并[4,3-a]吡啶-8-甲酰胺(I-5)Example 5: N-(2-Methyl-[1,1'-biphenyl]-3-yl)-6-(((2-sulfamoylethyl)amino)methyl)-[1,2 ,4]Triazolo[4,3-a]pyridine-8-carboxamide (I-5)
ESI-MS m/z:465.2[M+H]+；1H NMR(600MHz,DMSO-d6)δ11.40(s,1H),9.18(s,1H),8.75(s,1H),8.54(t,J＝14.7Hz,1H),8.21(t,J＝15.1Hz,1H),7.60–7.23(m,6H),7.09(d,J＝7.4Hz,1H),6.82(s,2H),3.93(s,2H),3.19(t,J＝7.1Hz,2H),2.92(t,J＝7.1Hz,2H),2.35(s,3H),1.23(s,1H).ESI-MS m/z: 465.2 [M+H] + ; 1 H NMR (600 MHz, DMSO-d 6 ) δ 11.40 (s, 1H), 9.18 (s, 1H), 8.75 (s, 1H), 8.54 (t, J=14.7Hz, 1H), 8.21 (t, J=15.1Hz, 1H), 7.60–7.23 (m, 6H), 7.09 (d, J=7.4Hz, 1H), 6.82 (s, 2H) ,3.93(s,2H),3.19(t,J=7.1Hz,2H),2.92(t,J=7.1Hz,2H),2.35(s,3H),1.23(s,1H).
实施例6：N-(2-甲基-[1,1'-联苯]-3-基)-6-(((2-(甲磺酰胺基)乙基)氨基)甲基)-[1,2,4]三唑并[4,3-a]吡啶-8-甲酰胺(I-6)Example 6: N-(2-Methyl-[1,1'-biphenyl]-3-yl)-6-(((2-(methylsulfonamido)ethyl)amino)methyl)-[ 1,2,4]Triazolo[4,3-a]pyridine-8-carboxamide (I-6)
ESI-MS m/z:479.2[M+H]+；ESI-MS m/z: 479.2 [M+H] + ;
实施例7：((8-((2-甲基-[1,1'-联苯]-3-基)氨甲酰基)-[1,2,4]三唑并[4,3-a]吡啶-6-基)甲基)-D-丙氨酸(I-7)Example 7: ((8-((2-Methyl-[1,1'-biphenyl]-3-yl)carbamoyl)-[1,2,4]triazolo[4,3-a ]pyridin-6-yl)methyl)-D-alanine (I-7)
ESI-MS m/z:430.2[M+H]+；ESI-MS m/z: 430.2 [M+H] + ;
实施例8：((8-((2-甲基-[1,1'-联苯]-3-基)氨甲酰基)-[1,2,4]三唑并[4,3-a]吡啶-6-基)甲基)-L-丙氨酸(I-8)Example 8: ((8-((2-Methyl-[1,1'-biphenyl]-3-yl)carbamoyl)-[1,2,4]triazolo[4,3-a ]pyridin-6-yl)methyl)-L-alanine (I-8)
ESI-MS m/z:430.2[M+H]+；ESI-MS m/z: 430.2 [M+H] + ;
实施例9：(S)-3-羟基-4-(((8-((2-甲基-[1,1'-联苯]-3-基)氨甲酰基)-[1,2,4]三唑并[4,3-a]吡啶-6-基)甲基)氨基)丁酸(I-9)Example 9: (S)-3-hydroxy-4-(((8-((2-methyl-[1,1'-biphenyl]-3-yl)carbamoyl)-[1,2, 4] Triazolo[4,3-a]pyridin-6-yl)methyl)amino)butyric acid (I-9)
ESI-MS m/z:460.2[M+H]+；ESI-MS m/z: 460.2[M+H] + ;
实施例10：N-(2-氰基-[1,1'-联苯]-3-基)-6-(((2-羟乙基)氨基)甲基)-[1,2,4]三唑并[4,3-a]吡啶-8-甲酰胺(I-10)Example 10: N-(2-cyano-[1,1'-biphenyl]-3-yl)-6-(((2-hydroxyethyl)amino)methyl)-[1,2,4 ]Triazolo[4,3-a]pyridine-8-carboxamide (I-10)
步骤1：3-氨基-[1,1'-联苯]-2-甲腈Step 1: 3-Amino-[1,1'-biphenyl]-2-carbonitrile
室温下，将2-氨基-6-溴苯甲腈(15.88g，0.081mol)、苯硼酸(12.2g，0.1mol)、醋酸钯(1.82g，8.1mmol)、碳酸钾(24.9g，0.18mol)加入至乙醇/水的混合溶液中(体积比1：1，100mL)，于N2保护下搅拌反应8小时。反映完毕后，抽滤，蒸干滤液，柱层析分离得白色固体12g，收率76.4％。At room temperature, 2-amino-6-bromobenzonitrile (15.88g, 0.081mol), phenylboronic acid (12.2g, 0.1mol), palladium acetate (1.82g, 8.1mmol), potassium carbonate (24.9g, 0.18mol) were mixed ) was added to the mixed solution of ethanol/water (volume ratio 1:1, 100 mL), and the reaction was stirred for 8 hours under the protection of N 2 . After the reaction was completed, suction filtration, the filtrate was evaporated to dryness, and 12 g of white solid was obtained by column chromatography, with a yield of 76.4%.
步骤2：6-溴-N-(2-氰基-[1,1'-联苯]-3-基)-[1,2,4]三唑并[4,3-a]吡啶-8-甲酰胺Step 2: 6-Bromo-N-(2-cyano-[1,1'-biphenyl]-3-yl)-[1,2,4]triazolo[4,3-a]pyridine-8 -Formamide
室温下，将6-溴-[1,2,4]三唑并[4,3-a]吡啶-8-甲酸(5g，0.021mol)、3-氨基-[1,1'-联苯]-2-甲腈(3.88g，0.02mol)、HATU(11.8g，0.031mol)、N，N-二异丙基乙胺(5.42g，0.042mol)溶于N,N-二甲基甲酰胺(50mL)中，搅拌反应12小时。反应完毕后，将反应液倒入大量水中，搅拌30分钟，抽滤，水洗滤饼，干燥，柱层析分离得黄色固体6.94g，收率为79.2％。6-Bromo-[1,2,4]triazolo[4,3-a]pyridine-8-carboxylic acid (5 g, 0.021 mol), 3-amino-[1,1'-biphenyl] -2-carbonitrile (3.88g, 0.02mol), HATU (11.8g, 0.031mol), N,N-diisopropylethylamine (5.42g, 0.042mol) in N,N-dimethylformamide (50 mL), the reaction was stirred for 12 hours. After the reaction was completed, the reaction solution was poured into a large amount of water, stirred for 30 minutes, suction filtered, the filter cake was washed with water, dried, and separated by column chromatography to obtain 6.94 g of a yellow solid with a yield of 79.2%.
步骤3：N-(2-氰基-[1,1'-联苯]-3-基)-6-乙烯基-[1,2,4]三唑并[4,3-a]吡啶-8-甲酰胺Step 3: N-(2-Cyano-[1,1'-biphenyl]-3-yl)-6-vinyl-[1,2,4]triazolo[4,3-a]pyridine- 8-Carboxamide
室温下，将6-溴-N-(2-氰基-[1,1'-联苯]-3-基)-[1,2,4]三唑并[4,3-a]吡啶-8-甲酰胺(2.38g，5.7mmol)、乙烯基硼酸频哪醇酯(0.96g，6.2mmol)、[1，1’-双(二苯基膦基)二茂铁]二氯化钯(0.42g，0.57mmol)、碳酸铯(3.4g，10.4mmol)溶于1，4-二氧六环/水(体积比3：1，20mL)的混合溶液中，N2保护下90℃反应3小时。反应液冷却至室温，用二氯甲烷萃取，蒸干有机层，柱层析分离得白色固体0.96g，收率为46.1％。At room temperature, 6-bromo-N-(2-cyano-[1,1'-biphenyl]-3-yl)-[1,2,4]triazolo[4,3-a]pyridine- 8-Carboxamide (2.38 g, 5.7 mmol), vinylboronic acid pinacol ester (0.96 g, 6.2 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride ( 0.42g, 0.57mmol) and cesium carbonate (3.4g, 10.4mmol) were dissolved in a mixed solution of 1,4-dioxane/water (volume ratio 3:1, 20mL), and reacted at 90°C under the protection of N2 for 3 Hour. The reaction solution was cooled to room temperature, extracted with dichloromethane, the organic layer was evaporated to dryness, and separated by column chromatography to obtain 0.96 g of a white solid with a yield of 46.1%.
步骤4：N-(2-氰基-[1,1'-联苯]-3-基)-6-甲酰基-[1,2,4]三唑并[4,3-a]吡啶-8-甲酰胺Step 4: N-(2-Cyano-[1,1'-biphenyl]-3-yl)-6-formyl-[1,2,4]triazolo[4,3-a]pyridine- 8-Carboxamide
室温下，将N-(2-氰基-[1,1'-联苯]-3-基)-6-乙烯基-[1,2,4]三唑并[4,3-a]吡啶-8-甲酰胺(0.66g，1.8mmol)溶于1，4-二氧六环/水(体积比5：1，15mL)的混合溶液中，快速加入2％的OsO4水溶液(3.5mL)，搅拌10分钟后加入高碘酸钠(1.54g，7.2mmol)，室温搅拌反应10小时。抽滤，得黄色固体0.51g，收率为77.2％。At room temperature, N-(2-cyano-[1,1'-biphenyl]-3-yl)-6-vinyl-[1,2,4]triazolo[4,3-a]pyridine -8-Carboxamide (0.66g, 1.8mmol) was dissolved in a mixed solution of 1,4-dioxane/water (volume ratio 5:1, 15mL), and a 2% OsO4 aqueous solution (3.5mL) was quickly added After stirring for 10 minutes, sodium periodate (1.54 g, 7.2 mmol) was added, and the reaction was stirred at room temperature for 10 hours. Suction filtration to obtain 0.51 g of a yellow solid with a yield of 77.2%.
步骤5：N-(2-氰基-[1,1'-联苯]-3-基)-6-(((2-羟乙基)氨基)甲基)-[1,2,4]三唑并[4,3-a]吡啶-8-甲酰胺(实施例10)Step 5: N-(2-Cyano-[1,1'-biphenyl]-3-yl)-6-(((2-hydroxyethyl)amino)methyl)-[1,2,4] Triazolo[4,3-a]pyridine-8-carboxamide (Example 10)
室温下，将N-(2-氰基-[1,1'-联苯]-3-基)-6-甲酰基-[1,2,4]三唑并[4,3-a]吡啶-8-甲酰胺(0.08g，0.22mmol)溶于DMF中，加入乙醇胺(0.067g，1.1mmol)、冰醋酸(0.02g，0.33mmol)，搅拌反应2小时。向体系中加入氰基硼氢化钠(0.07g，1.1mmol)，25℃搅拌反应12小时。反应液中加水，用二氯甲烷萃取，蒸干溶剂，柱层析分离得白色固体0.025g，收率为27.3％。At room temperature, N-(2-cyano-[1,1'-biphenyl]-3-yl)-6-formyl-[1,2,4]triazolo[4,3-a]pyridine -8-Carboxamide (0.08 g, 0.22 mmol) was dissolved in DMF, ethanolamine (0.067 g, 1.1 mmol) and glacial acetic acid (0.02 g, 0.33 mmol) were added, and the reaction was stirred for 2 hours. Sodium cyanoborohydride (0.07 g, 1.1 mmol) was added to the system, and the reaction was stirred at 25°C for 12 hours. Water was added to the reaction solution, extracted with dichloromethane, the solvent was evaporated to dryness, and 0.025 g of a white solid was obtained by column chromatography with a yield of 27.3%.
ESI-MS m/z:413.2[M+H]+；1H NMR(400MHz,DMSO-d6)δ11.42(s,1H),9.17(s,1H),8.75(s,1H),8.41(d,J＝1.2Hz,1H),8.19(d,J＝7.7Hz,1H),7.49(m,2H),7.47–7.16(m,4H),7.10(d,J＝7.6Hz,1H),4.67(s,1H),3.82(s,2H),3.55(d,J＝4.8Hz,2H),3.21(d,J＝4.7Hz,1H),2.67(m,2H).ESI-MS m/z: 413.2 [M+H] + ; 1 H NMR (400 MHz, DMSO-d 6 ) δ 11.42 (s, 1H), 9.17 (s, 1H), 8.75 (s, 1H), 8.41 (d, J=1.2Hz, 1H), 8.19 (d, J=7.7Hz, 1H), 7.49 (m, 2H), 7.47–7.16 (m, 4H), 7.10 (d, J=7.6Hz, 1H) ,4.67(s,1H),3.82(s,2H),3.55(d,J=4.8Hz,2H),3.21(d,J=4.7Hz,1H),2.67(m,2H).
根据实施例10的合成方法，以N-(2-氰基-[1,1'-联苯]-3-基)-6-甲酰基-[1,2,4]三唑并[4,3-a]吡啶-8-甲酰胺为原料，通过与不同的胺类化合物反应，再经氰基硼氢化钠或三乙酰氧基硼氢化钠还原制备得到实施例11-14的化合物。According to the synthetic method of Example 10, N-(2-cyano-[1,1'-biphenyl]-3-yl)-6-formyl-[1,2,4]triazolo[4, 3-a]pyridine-8-carboxamide is used as a raw material, and the compounds of Examples 11-14 are prepared by reacting with different amine compounds, and then reducing by sodium cyanoborohydride or sodium triacetoxyborohydride.
实施例11：((8-((2-氰基-[1,1'-联苯]-3-基)氨甲酰基)-[1,2,4]三唑并[4,3-a]吡啶-6-基)甲基)-D-丙氨酸(I-13)Example 11: ((8-((2-cyano-[1,1'-biphenyl]-3-yl)carbamoyl)-[1,2,4]triazolo[4,3-a ]pyridin-6-yl)methyl)-D-alanine (I-13)
ESI-MS m/z:441.2[M+H]+；ESI-MS m/z: 441.2 [M+H] + ;
实施例12：((8-((2-氰基-[1,1'-联苯]-3-基)氨甲酰基)-[1,2,4]三唑并[4,3-a]吡啶-6-基)甲基)-D-丝氨酸(I-14)Example 12: ((8-((2-cyano-[1,1'-biphenyl]-3-yl)carbamoyl)-[1,2,4]triazolo[4,3-a ]pyridin-6-yl)methyl)-D-serine (I-14)
ESI-MS m/z:457.2[M+H]+；ESI-MS m/z: 457.2 [M+H] + ;
实施例13：((8-((2-氰基-[1,1'-联苯]-3-基)氨甲酰基)-[1,2,4]三唑并[4,3-a]吡啶-6-基)甲基)-D-苏氨酸(I-15)Example 13: ((8-((2-Cyano-[1,1'-biphenyl]-3-yl)carbamoyl)-[1,2,4]triazolo[4,3-a ]pyridin-6-yl)methyl)-D-threonine (I-15)
ESI-MS m/z:471.2[M+H]+；ESI-MS m/z: 471.2 [M+H] + ;
实施例14：(S)-N-(氰基-[1,1'-联苯]-3-基)-6-((2-(羟甲基)吡咯烷-1-基)甲基)-[1,2,4]三唑并[4,3-a]吡啶-8-甲酰胺(I-16)Example 14: (S)-N-(Cyano-[1,1'-biphenyl]-3-yl)-6-((2-(hydroxymethyl)pyrrolidin-1-yl)methyl) -[1,2,4]Triazolo[4,3-a]pyridine-8-carboxamide (I-16)
ESI-MS m/z:453.2[M+H]+；ESI-MS m/z: 453.2 [M+H] + ;
实施例15：N-(3-(2,3-二氢苯并[b][1,4]二氧芑-6-基)-2-甲基苯基)-6-(((2-羟乙基)氨基)甲基)-[1,2,4]三唑并[4,3-a]吡啶-8-甲酰胺(I-17)Example 15: N-(3-(2,3-Dihydrobenzo[b][1,4]dioxan-6-yl)-2-methylphenyl)-6-(((2- Hydroxyethyl)amino)methyl)-[1,2,4]triazolo[4,3-a]pyridine-8-carboxamide (I-17)
步骤1：3-(2,3-二氢苯并[b][1,4]二氧芑-6-基)-2-甲基苯胺Step 1: 3-(2,3-Dihydrobenzo[b][1,4]dioxin-6-yl)-2-methylaniline
室温下，将3-溴-2-甲基苯胺(1.89g，10.21mmol)、2-(2,3-二氢苯并[b][1,4]二氧芑-6-基)-4,4,5,5-四甲基-1,3,2--二杂氧环戊硼烷(4.02g，15.32mmol)、[1,1-双(二苯基磷)二茂铁]二氯化钯(0.74g，1.02mmol)、碳酸钾(4.23g，30.63mmol)加入至二氧六环/水的混合溶液中(体积比2：1，30mL)，于N2保护下100度搅拌反应10小时。反映完毕后，抽滤，蒸干滤液，柱层析分离得白色固体1.53g，收率62.1％。At room temperature, 3-bromo-2-methylaniline (1.89 g, 10.21 mmol), 2-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)-4 ,4,5,5-tetramethyl-1,3,2-dioxaborolane (4.02g, 15.32mmol), [1,1-bis(diphenylphosphonium)ferrocene]bis Palladium chloride (0.74g, 1.02mmol), potassium carbonate (4.23g, 30.63mmol) were added to the mixed solution of dioxane/water (volume ratio 2 :1, 30mL), stirred at 100 degrees under the protection of N The reaction was carried out for 10 hours. After the reaction was completed, suction filtration, the filtrate was evaporated to dryness, and 1.53 g of white solid was obtained by column chromatography with a yield of 62.1%.
步骤2：6-溴-N-(3-(2,3-二氢苯并[b][1,4]二氧芑-6-基)-2-甲基苯基)-[1,2,4]三唑并[4,3-a]吡啶-8-甲酰胺Step 2: 6-Bromo-N-(3-(2,3-dihydrobenzo[b][1,4]dioxan-6-yl)-2-methylphenyl)-[1,2 ,4]Triazolo[4,3-a]pyridine-8-carboxamide
室温下，将6-溴-[1,2,4]三唑并[4,3-a]吡啶-8-甲酸(5g，0.021mol)、3-(2,3-二氢苯并[b][1,4]二氧芑-6-基)-2-甲基苯胺(5.06g，0.021mol)、HATU(11.8g，0.031mol)、N，N-二异丙基乙胺(5.4g，0.042mol)溶于N,N-二甲基甲酰胺(50mL)中，搅拌反应12小时。反应完毕后，将反应液倒入大量水中，搅拌30分钟，抽滤，水洗滤饼，干燥，柱层析分离得淡黄色固体8.7g，收率为89.3％。6-Bromo-[1,2,4]triazolo[4,3-a]pyridine-8-carboxylic acid (5 g, 0.021 mol), 3-(2,3-dihydrobenzo[b] ][1,4]dioxan-6-yl)-2-methylaniline (5.06g, 0.021mol), HATU (11.8g, 0.031mol), N,N-diisopropylethylamine (5.4g , 0.042 mol) was dissolved in N,N-dimethylformamide (50 mL), and the reaction was stirred for 12 hours. After the completion of the reaction, the reaction solution was poured into a large amount of water, stirred for 30 minutes, suction filtered, the filter cake was washed with water, dried, and separated by column chromatography to obtain 8.7 g of a pale yellow solid with a yield of 89.3%.
步骤3：N-(3-(2,3-二氢苯并[b][1,4]二氧芑-6-基)-2-甲基苯基)-6-乙烯基-[1,2,4]三唑并[4,3-a]吡啶-8-甲酰胺Step 3: N-(3-(2,3-Dihydrobenzo[b][1,4]dioxan-6-yl)-2-methylphenyl)-6-vinyl-[1, 2,4]Triazolo[4,3-a]pyridine-8-carboxamide
室温下，将6-溴-N-(3-(2,3-二氢苯并[b][1,4]二氧芑-6-基)-2-甲基苯基)-[1,2,4]三唑并[4,3-a]吡啶-8-甲酰胺(3g，6.5mmol)、乙烯基硼酸频哪醇酯(1.1g，7.2mmol)、[1，1’-双(二苯基膦基)二茂铁]二氯化钯(0.48g，0.65mmol)、碳酸铯(3.81g，11.7mmol)溶于1，4-二氧六环/水(体积比3：1，30mL)的混合溶液中，N2保护下90℃反应3小时。反应液冷却至室温，用二氯甲烷萃取，蒸干有机层，柱层析分离得黄色固体1.61g，收率为60.2％。At room temperature, 6-bromo-N-(3-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)-2-methylphenyl)-[1, 2,4]Triazolo[4,3-a]pyridine-8-carboxamide (3g, 6.5mmol), vinylboronic acid pinacol ester (1.1g, 7.2mmol), [1,1'-bis( Diphenylphosphino)ferrocene]palladium dichloride (0.48g, 0.65mmol), cesium carbonate (3.81g, 11.7mmol) were dissolved in 1,4-dioxane/water (volume ratio 3:1, 30 mL) of the mixed solution, reacted at 90 °C for 3 hours under the protection of N 2 . The reaction solution was cooled to room temperature, extracted with dichloromethane, the organic layer was evaporated to dryness, and separated by column chromatography to obtain 1.61 g of a yellow solid with a yield of 60.2%.
步骤4：N-(3-(2,3-二氢苯并[b][1,4]二氧芑-6-基)-2-甲基苯基)-6-甲酰基-[1,2,4]三唑并[4,3-a]吡啶-8-甲酰胺Step 4: N-(3-(2,3-Dihydrobenzo[b][1,4]dioxan-6-yl)-2-methylphenyl)-6-formyl-[1, 2,4]Triazolo[4,3-a]pyridine-8-carboxamide
室温下，将N-(3-(2,3-二氢苯并[b][1,4]二氧芑-6-基)-2-甲基苯基)-6-乙烯基-[1,2,4]三唑并[4,3-a]吡啶-8-甲酰胺(0.62g，1.5mmol)溶于1，4-二氧六环/水(体积比5：1，15mL)的混合溶液中，快速加入2％的OsO4水溶液(3mL)，搅拌10分钟后加入高碘酸钠(1.28g，6mmol)，室温搅拌反应8小时。抽滤，干燥，得类白色固体0.52g，收率为83.7％。At room temperature, N-(3-(2,3-dihydrobenzo[b][1,4]dioxan-6-yl)-2-methylphenyl)-6-vinyl-[1 ,2,4]Triazolo[4,3-a]pyridine-8-carboxamide (0.62 g, 1.5 mmol) was dissolved in 1,4-dioxane/water (volume ratio 5:1, 15 mL) To the mixed solution, a 2% OsO 4 aqueous solution (3 mL) was rapidly added, and after stirring for 10 minutes, sodium periodate (1.28 g, 6 mmol) was added, and the reaction was stirred at room temperature for 8 hours. Suction filtration and drying to obtain 0.52 g of an off-white solid with a yield of 83.7%.
步骤5：N-(3-(2,3-二氢苯并[b][1,4]二氧芑-6-基)-2-甲基苯基)-6-(((2-羟乙基)氨基)甲基)-[1,2,4]三唑并[4,3-a]吡啶-8-甲酰胺(实施例15)Step 5: N-(3-(2,3-Dihydrobenzo[b][1,4]dioxan-6-yl)-2-methylphenyl)-6-(((2-hydroxy Ethyl)amino)methyl)-[1,2,4]triazolo[4,3-a]pyridine-8-carboxamide (Example 15)
室温下，将N-(3-(2,3-二氢苯并[b][1,4]二氧芑-6-基)-2-甲基苯基)-6-甲酰基-[1,2,4]三唑并[4,3-a]吡啶-8-甲酰胺(0.09g，0.22mmol)溶于二氯甲烷中(8mL)，加入乙醇胺(0.067g，1.1mmol)、冰醋酸(0.02g，0.33mmol)，搅拌反应2小时。向体系中加入三乙酰氧基硼氢化钠(0.07g，1.1mmol)，25℃搅拌反应12小时。反应液中加水，用二氯甲烷萃取，蒸干溶剂，柱层析分离得白色固体0.04g，收率为17.5％。At room temperature, N-(3-(2,3-dihydrobenzo[b][1,4]dioxan-6-yl)-2-methylphenyl)-6-formyl-[1 ,2,4]Triazolo[4,3-a]pyridine-8-carboxamide (0.09g, 0.22mmol) was dissolved in dichloromethane (8mL), ethanolamine (0.067g, 1.1mmol), glacial acetic acid were added (0.02 g, 0.33 mmol) and the reaction was stirred for 2 hours. Sodium triacetoxyborohydride (0.07 g, 1.1 mmol) was added to the system, and the reaction was stirred at 25° C. for 12 hours. Water was added to the reaction solution, extracted with dichloromethane, the solvent was evaporated to dryness, and 0.04 g of a white solid was obtained by column chromatography with a yield of 17.5%.
ESI-MS m/z:460.2[M+H]+；1H NMR(600MHz,DMSO-d6)δ11.37(s,1H),9.16(s,1H),8.74(s,1H),8.52(s,1H),8.15(d,J＝7.9Hz,1H),7.30(t,J＝7.5Hz,1H),7.05(d,J＝7.4Hz,1H),6.93(d,J＝8.1Hz,1H),6.85–6.78(m,2H),4.52(s,1H),4.29(s,4H),3.92(s,2H),3.49(d,J＝4.9Hz,2H),2.61(t,J＝4.9Hz,2H),2.35(s,3H),1.23(s,1H).ESI-MS m/z: 460.2 [M+H] + ; 1 H NMR (600 MHz, DMSO-d 6 ) δ 11.37 (s, 1H), 9.16 (s, 1H), 8.74 (s, 1H), 8.52 (s, 1H), 8.15 (d, J=7.9Hz, 1H), 7.30 (t, J=7.5Hz, 1H), 7.05 (d, J=7.4Hz, 1H), 6.93 (d, J=8.1Hz) ,1H),6.85–6.78(m,2H),4.52(s,1H),4.29(s,4H),3.92(s,2H),3.49(d,J=4.9Hz,2H),2.61(t, J=4.9Hz, 2H), 2.35(s, 3H), 1.23(s, 1H).
根据实施例15的合成方法，以N-(3-(2,3-二氢苯并[b][1,4]二氧芑-6-基)-2-甲基苯基)-6-甲酰基-[1,2,4]三唑并[4,3-a]吡啶-8-甲酰胺为原料，通过与不同的胺类化合物反应，再经与三乙酰氧基硼氢化钠还原制备得到实施例16-21的化合物。According to the synthetic method of Example 15, N-(3-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)-2-methylphenyl)-6- Formyl-[1,2,4]triazolo[4,3-a]pyridine-8-carboxamide is prepared by reacting with different amine compounds and then reducing with sodium triacetoxyborohydride The compounds of Examples 16-21 were obtained.
实施例16：N-(3-(2,3-二氢苯并[b][1,4]二氧芑-6-基)-2-甲基苯基)-6-(((2-羟乙基)(甲基)氨基)甲基)-[1,2,4]三唑并[4,3-a]吡啶-8-甲酰胺(I-18)Example 16: N-(3-(2,3-Dihydrobenzo[b][1,4]dioxan-6-yl)-2-methylphenyl)-6-(((2- Hydroxyethyl)(methyl)amino)methyl)-[1,2,4]triazolo[4,3-a]pyridine-8-carboxamide (I-18)
ESI-MS m/z:474.2[M+H]+；1H NMR(600MHz,DMSO-d6)δ11.37(s,1H),9.20(s,1H),8.75(s,1H),8.47(s,1H),8.14(d,J＝7.8Hz,1H),7.30(t,J＝7.7Hz,1H),7.05(d,J＝7.4Hz,1H),6.93(d,J＝8.1Hz,1H),6.85–6.79(m,2H),4.50(s,1H),4.29(s,4H),3.72(s,2H),3.56(d,J＝5.5Hz,2H),2.35(s,3H),2.23(s,3H),1.23(s,2H).ESI-MS m/z: 474.2 [M+H] + ; 1 H NMR (600 MHz, DMSO-d 6 ) δ 11.37 (s, 1H), 9.20 (s, 1H), 8.75 (s, 1H), 8.47 (s, 1H), 8.14 (d, J=7.8Hz, 1H), 7.30 (t, J=7.7Hz, 1H), 7.05 (d, J=7.4Hz, 1H), 6.93 (d, J=8.1Hz) ,1H),6.85–6.79(m,2H),4.50(s,1H),4.29(s,4H),3.72(s,2H),3.56(d,J=5.5Hz,2H),2.35(s, 3H), 2.23(s, 3H), 1.23(s, 2H).
实施例17：6-(((2-乙酰胺基乙基)氨基)甲基)-N-(3-(2,3-二氢苯并[b][1,4]二氧芑-6-基)-2-甲基苯基)-[1,2,4]三唑并[4,3-a]吡啶-8-甲酰胺(I-19)Example 17: 6-(((2-acetamidoethyl)amino)methyl)-N-(3-(2,3-dihydrobenzo[b][1,4]dioxin-6 -yl)-2-methylphenyl)-[1,2,4]triazolo[4,3-a]pyridine-8-carboxamide (I-19)
ESI-MS m/z:501.2[M+H]+；1H NMR(600MHz,DMSO-d6)δ11.38(s,1H),9.16(s,1H),8.75(s,1H),8.53(s,1H),8.15(d,J＝8.0Hz,1H),7.82(s,1H),7.31(t,J＝7.8Hz,1H),7.06(d,J＝7.5Hz,1H),6.94(d,J＝8.2Hz,1H),6.85(d,J＝1.8Hz,1H),6.81(dd,J＝8.2,1.8Hz,1H),4.30(s,4H),3.91(s,2H),3.16(dd,J＝12.1,6.1Hz,2H),2.58(t,J＝6.4Hz,2H),2.36(s,3H),1.80(s,3H).ESI-MS m/z: 501.2 [M+H] + ; 1 H NMR (600 MHz, DMSO-d 6 ) δ 11.38 (s, 1H), 9.16 (s, 1H), 8.75 (s, 1H), 8.53 (s, 1H), 8.15 (d, J=8.0Hz, 1H), 7.82 (s, 1H), 7.31 (t, J=7.8Hz, 1H), 7.06 (d, J=7.5Hz, 1H), 6.94 (d,J＝8.2Hz,1H),6.85(d,J＝1.8Hz,1H),6.81(dd,J＝8.2,1.8Hz,1H),4.30(s,4H),3.91(s,2H) ,3.16(dd,J＝12.1,6.1Hz,2H),2.58(t,J＝6.4Hz,2H),2.36(s,3H),1.80(s,3H).
实施例18：N-(3-(2,3-二氢苯并[b][1,4]二氧芑-6-基)-2-甲基苯基)-6-(((2-(甲磺酰胺基)乙基)氨基)甲基)-[1,2,4]三唑并[4,3-a]吡啶-8-甲酰胺(I-20)Example 18: N-(3-(2,3-Dihydrobenzo[b][1,4]dioxan-6-yl)-2-methylphenyl)-6-(((2- (Methylsulfonamido)ethyl)amino)methyl)-[1,2,4]triazolo[4,3-a]pyridine-8-carboxamide (I-20)
ESI-MS m/z:537.2[M+H]+；1H NMR(600MHz,DMSO-d6)δ11.38(s,1H),9.18(s,1H),8.76(s,1H),8.53(s,1H),8.15(d,J＝8.0Hz,1H),7.31(t,J＝7.8Hz,1H),7.06(d,J＝7.5Hz,1H),6.98(s,1H),6.94(d,J＝8.2Hz,1H),6.85(d,J＝1.8Hz,1H),6.81(dd,J＝8.2,1.9Hz,1H),4.30(s,4H),3.92(s,2H),3.08(d,J＝5.3Hz,2H),2.92(s,3H),2.67(t,J＝6.3Hz,2H),2.36(s,3H).ESI-MS m/z: 537.2 [M+H] + ; 1 H NMR (600 MHz, DMSO-d 6 ) δ 11.38(s, 1H), 9.18(s, 1H), 8.76(s, 1H), 8.53 (s,1H),8.15(d,J=8.0Hz,1H),7.31(t,J=7.8Hz,1H),7.06(d,J=7.5Hz,1H),6.98(s,1H),6.94 (d, J=8.2Hz, 1H), 6.85 (d, J=1.8Hz, 1H), 6.81 (dd, J=8.2, 1.9Hz, 1H), 4.30 (s, 4H), 3.92 (s, 2H) ,3.08(d,J=5.3Hz,2H),2.92(s,3H),2.67(t,J=6.3Hz,2H),2.36(s,3H).
实施例19：((8-((3-(2,3-二氢苯并[b][1,4]二氧芑-6-基)-2-甲基苯基)氨甲酰基)-[1,2,4]三唑并[4,3-a]吡啶-6-基)甲基)-D-丙氨酸(I-21)Example 19: ((8-((3-(2,3-Dihydrobenzo[b][1,4]dioxin-6-yl)-2-methylphenyl)carbamoyl)- [1,2,4]Triazolo[4,3-a]pyridin-6-yl)methyl)-D-alanine (I-21)
ESI-MS m/z:488.2[M+H]+；ESI-MS m/z: 488.2 [M+H] + ;
实施例20：((8-((3-(2,3-二氢苯并[b][1,4]二氧芑-6-基)-2-甲基苯基)氨甲酰基)-[1,2,4]三唑并[4,3-a]吡啶-6-基)甲基)-L-丙氨酸(I-22)Example 20: ((8-((3-(2,3-Dihydrobenzo[b][1,4]dioxin-6-yl)-2-methylphenyl)carbamoyl)- [1,2,4]Triazolo[4,3-a]pyridin-6-yl)methyl)-L-alanine (I-22)
ESI-MS m/z:488.2[M+H]+；ESI-MS m/z: 488.2 [M+H] + ;
实施例21：(S)-4-(((8-((3-(2,3-二氢苯并[b][1,4]二氧芑-6-基)-2-甲基苯基)氨甲酰基)-[1,2,4]三唑并[4,3-a]吡啶-6-基)甲基)氨基)-3-羟基丁酸(I-23)Example 21: (S)-4-(((8-((3-(2,3-dihydrobenzo[b][1,4]dioxan-6-yl)-2-methylbenzene (I-23)
ESI-MS m/z:518.2[M+H]+；ESI-MS m/z: 518.2 [M+H] + ;
实施例22：N-(2-氰基-3-(2,3-二氢苯并[b][1,4]二氧芑-6-基)苯基)-6-(((2-羟乙基)氨基)甲基)-[1,2,4]三唑并[4,3-a]吡啶-8-甲酰胺(I-24)Example 22: N-(2-cyano-3-(2,3-dihydrobenzo[b][1,4]dioxan-6-yl)phenyl)-6-(((2- Hydroxyethyl)amino)methyl)-[1,2,4]triazolo[4,3-a]pyridine-8-carboxamide (I-24)
步骤1：2-氨基-6-(2,3-二氢苯并[b][1,4]二氧芑-6-基)苯甲腈Step 1: 2-Amino-6-(2,3-dihydrobenzo[b][1,4]dioxan-6-yl)benzonitrile
室温下，将2-氨基-6-溴苯甲腈(2g,10.2mmol)、2-(2,3-二氢苯并[b][1,4]二氧芑-6-基)-4,4,5,5-四甲基-1,3,2--二杂氧环戊硼烷(2.94g,11.22mmol)、四(三苯基膦)钯(1.18g,1.02mmol)、碳酸铯(6.65g,20.4mmol)加入至甲苯/乙醇/水的混合溶液中(体积比3：3：1，23mL)。在N2保护下，反应液于95℃搅拌反应12小时。反映完毕后，抽滤，蒸干滤液，柱层析分离得白色固体1.53g，收率为62.1％。At room temperature, 2-amino-6-bromobenzonitrile (2 g, 10.2 mmol), 2-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)-4 , 4,5,5-tetramethyl-1,3,2-dioxaborolane (2.94g, 11.22mmol), tetrakis(triphenylphosphine)palladium (1.18g, 1.02mmol), carbonic acid Cesium (6.65 g, 20.4 mmol) was added to a mixed solution of toluene/ethanol/water (volume ratio 3:3:1, 23 mL). Under the protection of N2 , the reaction solution was stirred at 95 °C for 12 hours. After the reaction was completed, suction filtration, the filtrate was evaporated to dryness, and 1.53 g of white solid was obtained by column chromatography, and the yield was 62.1%.
步骤2：6-溴-N-(2-氰基-3-(2,3-二氢苯并[b][1,4]二氧芑-6-基)苯基)-[1,2,4]三唑并[4,3-a]吡啶-8-甲酰胺Step 2: 6-Bromo-N-(2-cyano-3-(2,3-dihydrobenzo[b][1,4]dioxan-6-yl)phenyl)-[1,2 ,4]Triazolo[4,3-a]pyridine-8-carboxamide
室温下，将6-溴-[1,2,4]三唑并[4,3-a]吡啶-8-甲酸(1g，4.15mmol)溶于氯化亚砜中(15mL)，升温至80℃搅拌反应4小时。反应完毕后，蒸干反应液得黄色固体。将制得的黄色固体溶于二氯甲烷中(15mL)，冰浴条件下缓慢滴加到2-氨基-6-(2,3-二氢苯并[b][1,4]二氧芑-6-基)苯甲腈(1.05g，4.15mmol)和三乙胺(1.05g，10.38mmol)的二氯甲烷(10mL)溶液中，并于室温搅拌反应1小时。反应结束后，向反应液中加入大量二氯甲烷，平台搅拌1小时，加水洗涤有机层，蒸干有机层，得浅黄色固体。将粗品溶于乙醇中，80℃下搅拌半小时，抽滤，干燥得黄色固体0.89g，收率45.3％。At room temperature, 6-bromo-[1,2,4]triazolo[4,3-a]pyridine-8-carboxylic acid (1 g, 4.15 mmol) was dissolved in thionyl chloride (15 mL), and the temperature was raised to 80 The reaction was stirred for 4 hours. After the completion of the reaction, the reaction solution was evaporated to dryness to obtain a yellow solid. The obtained yellow solid was dissolved in dichloromethane (15 mL), and slowly added dropwise to 2-amino-6-(2,3-dihydrobenzo[b][1,4]dioxin under ice bath conditions) -6-yl)benzonitrile (1.05 g, 4.15 mmol) and triethylamine (1.05 g, 10.38 mmol) in dichloromethane (10 mL) and the reaction was stirred at room temperature for 1 hour. After the reaction, a large amount of dichloromethane was added to the reaction solution, the platform was stirred for 1 hour, the organic layer was washed with water, and the organic layer was evaporated to dryness to obtain a pale yellow solid. The crude product was dissolved in ethanol, stirred at 80° C. for half an hour, filtered with suction, and dried to obtain 0.89 g of a yellow solid with a yield of 45.3%.
步骤3：N-(2-氰基-3-(2,3-二氢苯并[b][1,4]二氧芑-6-基)苯基)-6-乙烯基-[1,2,4]三唑并[4,3-a]吡啶-8-甲酰胺Step 3: N-(2-cyano-3-(2,3-dihydrobenzo[b][1,4]dioxan-6-yl)phenyl)-6-vinyl-[1, 2,4]Triazolo[4,3-a]pyridine-8-carboxamide
室温下，将6-溴-N-(2-氰基-3-(2,3-二氢苯并[b][1,4]二氧芑-6-基)苯基)-[1,2,4]三唑并[4,3-a]吡啶-8-甲酰胺(3.09g，6.5mmol)、乙烯基硼酸频哪醇酯(1.1g，7.2mmol)、[1，1’-双(二苯基膦基)二茂铁]二氯化钯(0.48g，0.65mmol)、碳酸铯(3.81g，11.7mmol)溶于1，4-二氧六环/水(体积比3：1，30mL)的混合溶液中，N2保护下90℃反应3小时。反应液冷却至室温，用二氯甲烷萃取，蒸干有机层，柱层析分离得黄色固体1.75g，收率为63.7％。At room temperature, 6-bromo-N-(2-cyano-3-(2,3-dihydrobenzo[b][1,4]dioxan-6-yl)phenyl)-[1, 2,4]Triazolo[4,3-a]pyridine-8-carboxamide (3.09g, 6.5mmol), vinylboronic acid pinacol ester (1.1g, 7.2mmol), [1,1'-bis (diphenylphosphino)ferrocene]palladium dichloride (0.48g, 0.65mmol), cesium carbonate (3.81g, 11.7mmol) were dissolved in 1,4-dioxane/water (volume ratio 3:1 , 30 mL) of the mixed solution, reacted at 90 °C for 3 hours under the protection of N 2 . The reaction solution was cooled to room temperature, extracted with dichloromethane, the organic layer was evaporated to dryness, and 1.75 g of a yellow solid was obtained by column chromatography with a yield of 63.7%.
步骤4：N-(2-氰基-3-(2,3-二氢苯并[b][1,4]二氧芑-6-基)苯基)-6-甲酰基-[1,2,4]三唑并[4,3-a]吡啶-8-甲酰胺Step 4: N-(2-Cyano-3-(2,3-dihydrobenzo[b][1,4]dioxan-6-yl)phenyl)-6-formyl-[1, 2,4]Triazolo[4,3-a]pyridine-8-carboxamide
室温下，将N-(2-氰基-3-(2,3-二氢苯并[b][1,4]二氧芑-6-基)苯基)-6-乙烯基-[1,2,4]三唑并[4,3-a]吡啶-8-甲酰胺(0.63g，1.5mmol)溶于1，4-二氧六环/水(体积比5：1，15mL)的混合溶液中，快速加入2％的OsO4水溶液(3mL)，搅拌10分钟后加入高碘酸钠(1.28g，6mmol)，室温搅拌反应12小时。抽滤，干燥，得类白色固体0.58g，收率为90.2％。At room temperature, N-(2-cyano-3-(2,3-dihydrobenzo[b][1,4]dioxan-6-yl)phenyl)-6-vinyl-[1 ,2,4]Triazolo[4,3-a]pyridine-8-carboxamide (0.63 g, 1.5 mmol) was dissolved in 1,4-dioxane/water (volume ratio 5:1, 15 mL) To the mixed solution, a 2% OsO 4 aqueous solution (3 mL) was rapidly added, and after stirring for 10 minutes, sodium periodate (1.28 g, 6 mmol) was added, and the reaction was stirred at room temperature for 12 hours. Suction filtration and drying to obtain 0.58 g of an off-white solid with a yield of 90.2%.
步骤5：N-(2-氰基-3-(2,3-二氢苯并[b][1,4]二氧芑-6-基)苯基)-6-(((2-羟乙基)氨基)甲基)-[1,2,4]三唑并[4,3-a]吡啶-8-甲酰胺(实施例22)Step 5: N-(2-cyano-3-(2,3-dihydrobenzo[b][1,4]dioxan-6-yl)phenyl)-6-(((2-hydroxy Ethyl)amino)methyl)-[1,2,4]triazolo[4,3-a]pyridine-8-carboxamide (Example 22)
室温下，将N-(2-氰基-3-(2,3-二氢苯并[b][1,4]二氧芑-6-基)苯基)-6-甲酰基-[1,2,4]三唑并[4,3-a]吡啶-8-甲酰胺(0.094g，0.22mmol)溶于DMF中，加入乙醇胺(0.067g，1.1mmol)、冰醋酸(0.02g，0.33mmol)，搅拌反应2小时。向体系中加入氰基硼氢化钠(0.07g，1.1mmol)，25℃搅拌反应12小时。反应液中加水，用二氯甲烷萃取，蒸干溶剂，柱层析分离得白色固体0.029g，收率为27.6％。At room temperature, N-(2-cyano-3-(2,3-dihydrobenzo[b][1,4]dioxan-6-yl)phenyl)-6-formyl-[1 ,2,4]Triazolo[4,3-a]pyridine-8-carboxamide (0.094g, 0.22mmol) was dissolved in DMF, ethanolamine (0.067g, 1.1mmol), glacial acetic acid (0.02g, 0.33g) were added. mmol), and the reaction was stirred for 2 hours. Sodium cyanoborohydride (0.07 g, 1.1 mmol) was added to the system, and the reaction was stirred at 25°C for 12 hours. Water was added to the reaction solution, extracted with dichloromethane, the solvent was evaporated to dryness, and 0.029 g of a white solid was obtained by column chromatography with a yield of 27.6%.
ESI-MS m/z:471.2[M+H]+；1H NMR(600MHz,DMSO-d6)δ11.99(s,1H),9.21(s,1H),8.74(s,1H),8.55(s,1H),8.31(d,J＝8.2Hz,1H),7.80(t,J＝8.0Hz,1H),7.39(d,J＝7.6Hz,1H),7.14(s,1H),7.10(d,J＝7.5Hz,1H),7.03(d,J＝8.3Hz,1H),4.52(s,1H),4.33(s,4H),3.93(s,2H),3.50(d,J＝5.2Hz,2H),2.62(t,J＝5.5Hz,2H).ESI-MS m/z: 471.2 [M+H] + ; 1 H NMR (600 MHz, DMSO-d 6 ) δ 11.99 (s, 1H), 9.21 (s, 1H), 8.74 (s, 1H), 8.55 (s,1H),8.31(d,J=8.2Hz,1H),7.80(t,J=8.0Hz,1H),7.39(d,J=7.6Hz,1H),7.14(s,1H),7.10 (d, J=7.5Hz, 1H), 7.03(d, J=8.3Hz, 1H), 4.52(s, 1H), 4.33(s, 4H), 3.93(s, 2H), 3.50(d, J= 5.2Hz, 2H), 2.62(t, J=5.5Hz, 2H).
根据实施例22的合成方法，以N-(2-氰基-3-(2,3-二氢苯并[b][1,4]二氧芑-6-基)苯基)-6-甲酰基-[1,2,4]三唑并[4,3-a]吡啶-8-甲酰胺为原料，通过与不同的胺类化合物反应，再经与三乙酰氧基硼氢化钠还原制备得到实施例23-28的化合物。According to the synthetic method of Example 22, N-(2-cyano-3-(2,3-dihydrobenzo[b][1,4]dioxan-6-yl)phenyl)-6- Formyl-[1,2,4]triazolo[4,3-a]pyridine-8-carboxamide is prepared by reacting with different amine compounds and then reducing with sodium triacetoxyborohydride The compounds of Examples 23-28 were obtained.
实施例23：N-(2氰基-3-(2,3-二氢苯并[b][1,4]二氧芑-6-基)苯基)-6-(((2-羟乙基)(甲基)氨基)甲基)-[1,2,4]三唑并[4,3-a]吡啶-8-甲酰胺(I-25)Example 23: N-(2cyano-3-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)phenyl)-6-(((2-hydroxyl Ethyl)(methyl)amino)methyl)-[1,2,4]triazolo[4,3-a]pyridine-8-carboxamide (I-25)
ESI-MS m/z:485.2[M+H]+；ESI-MS m/z: 485.2 [M+H] + ;
实施例24：6-(((2-乙酰胺基乙基)氨基)甲基)-N-(2-氰基-3-(2,3-二氢苯并[b][1,4]二氧芑-6-基)苯基)-[1,2,4]三唑并[4,3-a]吡啶-8-甲酰胺(I-26)Example 24: 6-(((2-acetamidoethyl)amino)methyl)-N-(2-cyano-3-(2,3-dihydrobenzo[b][1,4]) Dioxan-6-yl)phenyl)-[1,2,4]triazolo[4,3-a]pyridine-8-carboxamide (I-26)
ESI-MS m/z:512.2[M+H]+；1H NMR(600MHz,DMSO-d6)δ11.99(s,1H),9.20(s,1H),8.75(s,1H),8.54(s,1H),8.32(s,1H),7.82(s,2H),7.40(s,1H),7.23–7.02(m,3H),4.33(s,4H),3.91(s,2H),3.16(s,2H),2.58(s,2H),1.80(s,3H).ESI-MS m/z: 512.2 [M+H] + ; 1 H NMR (600 MHz, DMSO-d 6 ) δ 11.99 (s, 1H), 9.20 (s, 1H), 8.75 (s, 1H), 8.54 (s, 1H), 8.32(s, 1H), 7.82(s, 2H), 7.40(s, 1H), 7.23–7.02(m, 3H), 4.33(s, 4H), 3.91(s, 2H), 3.16(s, 2H), 2.58(s, 2H), 1.80(s, 3H).
实施例25：N-(2-氰基-3-(2,3-二氢苯并[b][1,4]二氧芑-6-基)苯基)-6-(((2-(甲磺酰胺基)乙基)氨基)甲基)-[1,2,4]三唑并[4,3-a]吡啶-8-甲酰胺(I-27)Example 25: N-(2-cyano-3-(2,3-dihydrobenzo[b][1,4]dioxan-6-yl)phenyl)-6-(((2- (Methylsulfonamido)ethyl)amino)methyl)-[1,2,4]triazolo[4,3-a]pyridine-8-carboxamide (I-27)
ESI-MS m/z:548.2[M+H]+；ESI-MS m/z: 548.2 [M+H] + ;
实施例26：((8-((2-氰基-3-(2,3-二氢苯并[b][1,4]二氧芑-6-基)苯基)氨甲酰基)-[1,2,4]三唑并[4,3-a]吡啶-6-基)甲基)-D-丙氨酸(I-28)Example 26: ((8-((2-cyano-3-(2,3-dihydrobenzo[b][1,4]dioxan-6-yl)phenyl)carbamoyl)- [1,2,4]Triazolo[4,3-a]pyridin-6-yl)methyl)-D-alanine (I-28)
ESI-MS m/z:499.2[M+H]+；ESI-MS m/z: 499.2 [M+H] + ;
实施例27：((8-((2-氰基-3-(2,3-二氢苯并[b][1,4]二氧芑-6-基)苯基)氨甲酰基)-[1,2,4]三唑并[4,3-a]吡啶-6-基)甲基)-L-丙氨酸(I-29)Example 27: ((8-((2-cyano-3-(2,3-dihydrobenzo[b][1,4]dioxan-6-yl)phenyl)carbamoyl)- [1,2,4]Triazolo[4,3-a]pyridin-6-yl)methyl)-L-alanine (I-29)
ESI-MS m/z:499.2[M+H]+；ESI-MS m/z: 499.2 [M+H] + ;
实施例28：(S)-4-(((8-((2-氰基-3-(2,3-二氢苯并[b][1,4]二氧芑-6-基)苯基)氨甲酰基)-[1,2,4]三唑并[4,3-a]吡啶-6-基)甲基)氨基)-3-羟基丁酸(I-30)Example 28: (S)-4-(((8-((2-cyano-3-(2,3-dihydrobenzo[b][1,4]dioxan-6-yl)benzene yl)carbamoyl)-[1,2,4]triazolo[4,3-a]pyridin-6-yl)methyl)amino)-3-hydroxybutyric acid (I-30)
ESI-MS m/z:529.2[M+H]+；ESI-MS m/z: 529.2 [M+H] + ;
按照实施例15的制备方法，在第一步中采用3-溴-2-氯苯胺代替3-溴-2-甲基苯胺，与2-(2,3-二氢苯并[b][1,4]二氧芑-6-基)-4,4,5,5-四甲基-1,3,2-二杂氧环戊硼烷经过Suzuki偶联反应制得2-氯-3-(2,3-二氢苯并[b][1,4]二氧芑-6-基)苯胺，随后先后经过与6-溴-[1,2,4]三唑并[4,3-a]吡啶-8-甲酸酰胺化、乙烯基硼酸频哪醇酯偶联、高碘酸钠氧化、胺类化合物还原胺化制得实施例29-31的化合物。According to the preparation method of Example 15, in the first step, 3-bromo-2-chloroaniline was used instead of 3-bromo-2-methylaniline, and 2-(2,3-dihydrobenzo[b][1 ,4]dioxan-6-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane via Suzuki coupling to obtain 2-chloro-3- (2,3-Dihydrobenzo[b][1,4]dioxin-6-yl)aniline, followed by 6-bromo-[1,2,4]triazolo[4,3- a] Pyridine-8-carboxylic acid amidation, vinylboronic acid pinacol ester coupling, sodium periodate oxidation, and reductive amination of amine compounds to obtain the compounds of Examples 29-31.
实施例29：6-(((2-乙酰胺基乙基)氨基)甲基)-N-(2-氯-3-(2,3-二氢苯并[b][1,4]二氧芑-6-基)苯基)-[1,2,4]三唑并[4,3-a]吡啶-8-甲酰胺(I-33)Example 29: 6-(((2-acetamidoethyl)amino)methyl)-N-(2-chloro-3-(2,3-dihydrobenzo[b][1,4]di Oxygen-6-yl)phenyl)-[1,2,4]triazolo[4,3-a]pyridine-8-carboxamide (I-33)
ESI-MS m/z:521.2[M+H]+；ESI-MS m/z: 521.2 [M+H] + ;
实施例30：((8-((2-氯-3-(2,3-二氢苯并[b][1,4]二氧芑-6-基)苯基)氨甲酰基)-[1,2,4]三唑并[4,3-a]吡啶-6-基)甲基)-D-丙氨酸(I-34)Example 30: ((8-((2-Chloro-3-(2,3-dihydrobenzo[b][1,4]dioxan-6-yl)phenyl)carbamoyl)-[ 1,2,4]Triazolo[4,3-a]pyridin-6-yl)methyl)-D-alanine (I-34)
ESI-MS m/z:508.1[M+H]+；ESI-MS m/z: 508.1 [M+H] + ;
实施例31：(S)-1-((8-((2-氯-3-(2,3-二氢苯并[b][1,4]二氧芑-6-基)苯基)氨甲酰基)-[1,2,4]三唑并[4,3-a]吡啶-6-基)甲基)哌啶-2-甲酸(I-37)Example 31: (S)-1-((8-((2-Chloro-3-(2,3-dihydrobenzo[b][1,4]dioxan-6-yl)phenyl) Carbamoyl)-[1,2,4]triazolo[4,3-a]pyridin-6-yl)methyl)piperidine-2-carboxylic acid (I-37)
ESI-MS m/z:548.2[M+H]+；ESI-MS m/z: 548.2 [M+H] + ;
实施例32：6-(((2-羟乙基)氨基)甲基)-N-(2-甲基-3-(1-甲基-1H-吲唑-4-基)苯基)-[1,2,4]三唑并[4,3-a]吡啶-8-甲酰胺(I-38)Example 32: 6-(((2-hydroxyethyl)amino)methyl)-N-(2-methyl-3-(1-methyl-1H-indazol-4-yl)phenyl)- [1,2,4]Triazolo[4,3-a]pyridine-8-carboxamide (I-38)
步骤1：2-甲基-3-(1-甲基-1H-吲唑-4-基)苯胺Step 1: 2-Methyl-3-(1-methyl-1H-indazol-4-yl)aniline
室温下，将3-溴-2-甲基苯胺(12g，0.065mol)、1-甲基-4-(4,4,5,5-四甲基-1,3,2-二杂氧环戊硼烷-2-基)-1H-吲唑(33.5g，0.13mol)、醋酸钯(1.46g，6.5mmol)、碳酸钾(26.9g，0.195mol)加入至乙醇/水的混合溶液中(体积比1：1，200mL)，于N2保护下搅拌反应6小时。反映完毕后，抽滤，蒸干滤液，柱层析分离得白色固体9.85g，收率63.9％。At room temperature, 3-bromo-2-methylaniline (12 g, 0.065 mol), 1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxa Pentaboran-2-yl)-1H-indazole (33.5g, 0.13mol), palladium acetate (1.46g, 6.5mmol), potassium carbonate (26.9g, 0.195mol) were added to a mixed solution of ethanol/water ( Volume ratio 1:1, 200 mL), the reaction was stirred under N2 protection for 6 hours. After the reaction was completed, suction filtration, the filtrate was evaporated to dryness, and 9.85 g of a white solid was obtained by column chromatography with a yield of 63.9%.
步骤2：6-溴-N-(2-甲基-3-(1-甲基-1H-吲唑-4-基)苯基)-[1,2,4]三唑并[4,3-a]吡啶-8-甲酰胺Step 2: 6-Bromo-N-(2-methyl-3-(1-methyl-1H-indazol-4-yl)phenyl)-[1,2,4]triazolo[4,3 -a]pyridine-8-carboxamide
室温下，将6-溴-[1,2,4]三唑并[4,3-a]吡啶-8-甲酸(10g，0.042mol)、2-甲基-3-(1-甲基-1H-吲唑-4-基)苯胺(9.49g，0.04mol)、HATU(22.8g，0.06mol)、N，N-二异丙基乙胺(10.3g，0.08mol)溶于N，N-二甲基甲酰胺(20mL)中，搅拌反应13小时。反应完毕后，将反应液倒入大量水中，搅拌30分钟，抽滤，水洗滤饼，干燥，柱层析分离得浅黄色固体13.7g，收率为71.9％。6-Bromo-[1,2,4]triazolo[4,3-a]pyridine-8-carboxylic acid (10 g, 0.042 mol), 2-methyl-3-(1-methyl- 1H-Indazol-4-yl)aniline (9.49g, 0.04mol), HATU (22.8g, 0.06mol), N,N-diisopropylethylamine (10.3g, 0.08mol) dissolved in N,N- The reaction was stirred in dimethylformamide (20 mL) for 13 hours. After the reaction was completed, the reaction solution was poured into a large amount of water, stirred for 30 minutes, suction filtered, the filter cake was washed with water, dried, and separated by column chromatography to obtain 13.7 g of a light yellow solid with a yield of 71.9%.
步骤3：N-(2-甲基-3-(1-甲基-1H-吲唑-4-基)苯基)-6-乙烯基-[1,2,4]三唑并[4,3-a]吡啶-8-甲酰胺Step 3: N-(2-Methyl-3-(1-methyl-1H-indazol-4-yl)phenyl)-6-vinyl-[1,2,4]triazolo[4, 3-a]pyridine-8-carboxamide
室温下，将6-溴-N-(2-甲基-3-(1-甲基-1H-吲唑-4-基)苯基)-[1,2,4]三唑并[4,3-a]吡啶-8-甲酰胺(2g，4.3mmol)、乙烯基硼酸频哪醇酯(0.73g，4.7mmol)、[1，1’-双(二苯基膦基)二茂铁]二氯化钯(0.31g，0.43mmol)、碳酸铯(2.5g，7.7mmol)溶于1，4-二氧六环/水(体积比3：1，15mL)的混合溶液中，N2保护下90℃反应5小时。反应液冷却至室温，用二氯甲烷萃取，蒸干有机层，柱层析分离得白色固体1.19g，收率为67.7％。At room temperature, 6-bromo-N-(2-methyl-3-(1-methyl-1H-indazol-4-yl)phenyl)-[1,2,4]triazolo[4, 3-a]pyridine-8-carboxamide (2g, 4.3mmol), vinylboronic acid pinacol ester (0.73g, 4.7mmol), [1,1'-bis(diphenylphosphino)ferrocene] Palladium dichloride (0.31 g, 0.43 mmol), cesium carbonate (2.5 g, 7.7 mmol) were dissolved in a mixed solution of 1,4-dioxane/water (volume ratio 3:1, 15 mL), N2 protected The reaction was carried out at 90°C for 5 hours. The reaction solution was cooled to room temperature, extracted with dichloromethane, the organic layer was evaporated to dryness, and 1.19 g of a white solid was obtained by column chromatography with a yield of 67.7%.
步骤4：6-甲酰基-N-(2-甲基-3-(1-甲基-1H-吲唑-4-基)苯基)-[1,2,4]三唑并[4,3-a]吡啶-8-甲酰胺Step 4: 6-Formyl-N-(2-methyl-3-(1-methyl-1H-indazol-4-yl)phenyl)-[1,2,4]triazolo[4, 3-a]pyridine-8-carboxamide
室温下，将N-(2-甲基-3-(1-甲基-1H-吲唑-4-基)苯基)-6-乙烯基-[1,2,4]三唑并[4,3-a]吡啶-8-甲酰胺(0.73g，1.8mmol)溶于1，4-二氧六环/水(体积比5：1，8mL)的混合溶液中，快速加入2％的OsO4水溶液(3.5mL)，搅拌5分钟后加入高碘酸钠(1.54g，7.2mml)，室温搅拌反应5小时。抽滤，得淡黄色固体0.53g，收率为72.3％。At room temperature, N-(2-methyl-3-(1-methyl-1H-indazol-4-yl)phenyl)-6-vinyl-[1,2,4]triazolo[4 ,3-a]pyridine-8-carboxamide (0.73g, 1.8mmol) was dissolved in a mixed solution of 1,4-dioxane/water (volume ratio 5:1, 8mL), and 2% OsO was added rapidly 4 aqueous solution (3.5 mL), stirred for 5 minutes, added sodium periodate (1.54 g, 7.2 mml), and stirred at room temperature for 5 hours. Suction filtration to obtain 0.53 g of a pale yellow solid with a yield of 72.3%.
步骤5：6-(((2-羟乙基)氨基)甲基)-N-(2-甲基-3-(1-甲基-1H-吲唑-4-基)苯基)-[1,2,4]三唑并[4,3-a]吡啶-8-甲酰胺(实施例32)Step 5: 6-(((2-hydroxyethyl)amino)methyl)-N-(2-methyl-3-(1-methyl-1H-indazol-4-yl)phenyl)-[ 1,2,4]Triazolo[4,3-a]pyridine-8-carboxamide (Example 32)
室温下，将6-甲酰基-N-(2-甲基-3-(1-甲基-1H-吲唑-4-基)苯基)-[1,2,4]三唑并[4,3-a]吡啶-8-甲酰胺(0.09g，0.22mmol)溶于DMF中，加入乙醇胺(0.067g，1.1mmol)、冰醋酸(0.02g，0.33mmol)，搅拌反应2小时。向体系中加入氰基硼氢化钠(0.07g，1.1mmol)，25℃搅拌反应15小时。反应液中加水，用二氯甲烷萃取，蒸干溶剂，柱层析分离得白色固体0.02g，收率为20.1％。At room temperature, 6-formyl-N-(2-methyl-3-(1-methyl-1H-indazol-4-yl)phenyl)-[1,2,4]triazolo[4 ,3-a]pyridine-8-carboxamide (0.09g, 0.22mmol) was dissolved in DMF, ethanolamine (0.067g, 1.1mmol) and glacial acetic acid (0.02g, 0.33mmol) were added, and the reaction was stirred for 2 hours. Sodium cyanoborohydride (0.07 g, 1.1 mmol) was added to the system, and the reaction was stirred at 25°C for 15 hours. Water was added to the reaction solution, extracted with dichloromethane, the solvent was evaporated to dryness, and 0.02 g of a white solid was obtained by column chromatography with a yield of 20.1%.
根据实施例32的合成方法，以6-甲酰基-N-(2-甲基-3-(1-甲基-1H-吲唑-4-基)苯基)-[1,2,4]三唑并[4,3-a]吡啶-8-甲酰胺为原料，通过与不同的胺类化合物反应，再经氰基硼氢化钠还原制备得到实施例33和34的化合物。According to the synthetic method of Example 32, 6-formyl-N-(2-methyl-3-(1-methyl-1H-indazol-4-yl)phenyl)-[1,2,4] Triazolo[4,3-a]pyridine-8-carboxamide was used as raw material, and the compounds of Examples 33 and 34 were prepared by reacting with different amine compounds and reducing them with sodium cyanoborohydride.
实施例33：6-(((2-乙酰胺基乙基)氨基)甲基)-N-(2-甲基-3-(1-甲基-1H-吲唑-4-基)苯基)-[1,2,4]三唑并[4,3-a]吡啶-8-甲酰胺(I-40)Example 33: 6-(((2-acetamidoethyl)amino)methyl)-N-(2-methyl-3-(1-methyl-1H-indazol-4-yl)phenyl )-[1,2,4]Triazolo[4,3-a]pyridine-8-carboxamide (I-40)
ESI-MS m/z:497.2[M+H]+；ESI-MS m/z: 497.2 [M+H] + ;
实施例34：((8-((2-甲基-3-(1-甲基-1H-吲唑-4-基)苯基)氨甲酰基)-[1,2,4]三唑并[4,3-a]吡啶-6-基)甲基)-D-丙氨酸(I-41)Example 34: ((8-((2-Methyl-3-(1-methyl-1H-indazol-4-yl)phenyl)carbamoyl)-[1,2,4]triazolo [4,3-a]pyridin-6-yl)methyl)-D-alanine (I-41)
ESI-MS m/z:484.2[M+H]+；ESI-MS m/z: 484.2 [M+H] + ;
按照实施例1的制备方法，在第一步中采用取代的苯硼酸代替苯硼酸，与3-溴-2-甲基苯胺经过Suzuki偶联反应制得联苯胺类衍生物，随后先后经过与6-溴-[1,2,4]三唑并[4,3-a]吡啶-8-甲酸酰胺化、乙烯基硼酸频哪醇酯偶联、高碘酸钠氧化、乙醇胺还原胺化制得实施例35-39的化合物。According to the preparation method of Example 1, in the first step, substituted phenylboronic acid was used to replace phenylboronic acid, and benzidine derivatives were obtained through Suzuki coupling reaction with 3-bromo-2-methylaniline, and then successively with 6 -Bromo-[1,2,4]triazolo[4,3-a]pyridine-8-carboxylic acid amidation, vinylboronic acid pinacol ester coupling, sodium periodate oxidation, ethanolamine reductive amination Compounds of Examples 35-39.
实施例35：N-(2,4'-二甲基-[1,1'-联苯]-3-基)-6-(((2-羟乙基)氨基)甲基)-[1,2,4]三唑并[4,3-a]吡啶-8-甲酰胺(I-42)Example 35: N-(2,4'-Dimethyl-[1,1'-biphenyl]-3-yl)-6-(((2-hydroxyethyl)amino)methyl)-[1 ,2,4]Triazolo[4,3-a]pyridine-8-carboxamide (I-42)
ESI-MS m/z:416.2[M+H]+；ESI-MS m/z: 416.2 [M+H] + ;
实施例36：N-(2'-氟-3'-甲氧基-2-甲基-[1,1'-联苯]-3-基)-6-(((2-羟乙基)氨基)甲基)-[1,2,4]三唑并[4,3-a]吡啶-8-甲酰胺(I-43)Example 36: N-(2'-Fluoro-3'-methoxy-2-methyl-[1,1'-biphenyl]-3-yl)-6-(((2-hydroxyethyl) Amino)methyl)-[1,2,4]triazolo[4,3-a]pyridine-8-carboxamide (I-43)
ESI-MS m/z:450.2[M+H]+；ESI-MS m/z: 450.2 [M+H] + ;
实施例37：N-(3'-溴-2-甲基-[1,1'-联苯]-3-基)-6-(((2-羟乙基)氨基)甲基)-[1,2,4]三唑并[4,3-a]吡啶-8-甲酰胺(I-44)Example 37: N-(3'-Bromo-2-methyl-[1,1'-biphenyl]-3-yl)-6-(((2-hydroxyethyl)amino)methyl)-[ 1,2,4]Triazolo[4,3-a]pyridine-8-carboxamide (I-44)
ESI-MS m/z:480.1[M+H]+；ESI-MS m/z: 480.1 [M+H] + ;
实施例38：N-(4'-氯-2-甲基-[1,1'-联苯]-3-基)-6-(((2-羟乙基)氨基)甲基)-[1,2,4]三唑并[4,3-a]吡啶-8-甲酰胺(I-45)Example 38: N-(4'-Chloro-2-methyl-[1,1'-biphenyl]-3-yl)-6-(((2-hydroxyethyl)amino)methyl)-[ 1,2,4]Triazolo[4,3-a]pyridine-8-carboxamide (I-45)
ESI-MS m/z:436.2[M+H]+；ESI-MS m/z: 436.2[M+H] + ;
实施例39：N-(3',4'-二甲氧基-2-甲基-[1,1'-联苯]-3-基)-6-(((2-羟乙基)氨基)甲基)-[1,2,4]三唑并[4,3-a]吡啶-8-甲酰胺(I-46)Example 39: N-(3',4'-Dimethoxy-2-methyl-[1,1'-biphenyl]-3-yl)-6-(((2-hydroxyethyl)amino )methyl)-[1,2,4]triazolo[4,3-a]pyridine-8-carboxamide (I-46)
ESI-MS m/z:462.2[M+H]+；ESI-MS m/z: 462.2[M+H] + ;
本发明化合物生物学活性研究Study on the biological activity of the compounds of the present invention
采用均相时间分辨荧光(Homogenouse Time-Resolved Fluorescence，HTRF)试验来检测本发明化合物抑制PD-1/PD-L1相互作用的能力。检测试剂盒购买于CisBio公司(CAT#63ADK000CPAPEG)，其中包含Anti-Tag1-Cyptate、Anti-Tag2-XL665/d2、Tag1-PD-L1、Tag2-PD-1、Dilution Buffer、Detection Buffer等实验所需试剂。Homogenouse Time-Resolved Fluorescence (HTRF) assay was used to detect the ability of the compounds of the present invention to inhibit the interaction of PD-1/PD-L1. The detection kit was purchased from CisBio (CAT#63ADK000CPAPEG), which contains Anti-Tag1-Cyptate, Anti-Tag2-XL665/d2, Tag1-PD-L1, Tag2-PD-1, Dilution Buffer, Detection Buffer and other experiments required reagents.
实验步骤：PD-1重组蛋白和PD-L1重组蛋白分别用Dilution Buffer稀释至500nM和50nM。用Dilution Buffer稀释4mM用DMSO溶解的小分子化合物20倍至200uM。用含5％DMSO的Dilution Buffer四倍梯度稀释。同时用Dilution Buffer稀释600uM用DMSO溶解的PD-1/PD-L1抑制剂20倍至30uM，用含5％DMSO的Dilution Buffer四倍梯度稀释。依次向384孔中加入2uL稀释好的待测化合物、4uL稀释好的PD-1和4uL稀释好的PDL-1。充分混匀，室温放置15min。用Detection buffer稀释anti-Tag1-Eu3+(1:25)和anti-Tag2-XL665(1:100)。然后等体积混合稀释好的检测试剂，每反应孔加入10μL抗体混合液。封膜室温孵育2h。用ENVISION(Perkinelmer)仪器检测荧光信号(320nm刺激,665nm，615nm发射)。每个化合物检测8-12个浓度。Experimental steps: PD-1 recombinant protein and PD-L1 recombinant protein were diluted to 500nM and 50nM with Dilution Buffer, respectively. Dilute 4mM small molecule compounds dissolved in DMSO 20-fold to 200uM with Dilution Buffer. Four-fold serial dilutions were made with Dilution Buffer containing 5% DMSO. At the same time, dilute 600 uM of PD-1/PD-L1 inhibitor dissolved in DMSO by 20-fold to 30 uM with Dilution Buffer, and use four-fold gradient dilution with Dilution Buffer containing 5% DMSO. Add 2uL of diluted test compound, 4uL of diluted PD-1 and 4uL of diluted PDL-1 to 384 wells in sequence. Mix well and leave at room temperature for 15min. Dilute anti-Tag1-Eu 3+ (1:25) and anti-Tag2-XL665 (1:100) with Detection buffer. Then, equal volumes of the diluted detection reagents were mixed, and 10 μL of antibody mixture was added to each reaction well. The membrane was incubated at room temperature for 2 h. Fluorescence signals (320 nm excitation, 665 nm, 615 nm emission) were detected with an ENVISION (Perkinelmer) instrument. Each compound was tested at 8-12 concentrations.
化合物抑制PD-1/PD-L1相互作用的活性结果见表1。The activity results of compounds inhibiting PD-1/PD-L1 interaction are shown in Table 1.
表1本发明化合物抑制PD-1/PD-L1相互作用的活性范围或IC50。Table 1 Activity range or IC50 of the compounds of the present invention for inhibiting PD-1/PD-L1 interaction.
范围如下：A＝1nM-100nM；B＝100.01nM-1μM；C＝1.01μM-20μM。The ranges are as follows: A = 1 nM - 100 nM; B = 100.01 nM - 1 μM; C = 1.01 μM - 20 μM.
HTRF测试结果表明，实施例化合物在分子水平可显著抑制PD-1/PD-L1相互作用。有望在治疗与PD-1/PD-L1相互作用有关的疾病中展示出积极的作用。The HTRF test results showed that the example compounds could significantly inhibit the PD-1/PD-L1 interaction at the molecular level. It is expected to show positive effects in the treatment of diseases related to PD-1/PD-L1 interaction.
Claims (17)
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910200914.5A CN109897036B (en) | 2019-03-15 | 2019-03-15 | Triazolopyridine compounds and preparation method and use thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910200914.5A CN109897036B (en) | 2019-03-15 | 2019-03-15 | Triazolopyridine compounds and preparation method and use thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN109897036A CN109897036A (en) | 2019-06-18 |
| CN109897036B true CN109897036B (en) | 2021-07-30 |
Family
ID=66952268
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910200914.5A Active CN109897036B (en) | 2019-03-15 | 2019-03-15 | Triazolopyridine compounds and preparation method and use thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109897036B (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114805348B (en) * | 2022-05-20 | 2023-05-23 | 贵州大学 | A kind of triazolopyridine amide compound and its preparation method and application |
| CN116283971B (en) * | 2023-03-13 | 2024-04-12 | 沈阳药科大学 | Indoline compound containing condensed heterocyclic structure, and preparation method and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105440135A (en) * | 2014-09-01 | 2016-03-30 | 博笛生物科技有限公司 | Anti-PD-L1 conjugates for the treatment of tumors |
| WO2018119236A1 (en) * | 2016-12-22 | 2018-06-28 | Incyte Corporation | Triazolo[1,5-a]pyridine derivatives as immunomodulators |
| CN109195602A (en) * | 2016-08-03 | 2019-01-11 | 亚星国际公司 | Symmetrical or semisymmetrical compounds useful as immunomodulators |
-
2019
- 2019-03-15 CN CN201910200914.5A patent/CN109897036B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105440135A (en) * | 2014-09-01 | 2016-03-30 | 博笛生物科技有限公司 | Anti-PD-L1 conjugates for the treatment of tumors |
| CN109195602A (en) * | 2016-08-03 | 2019-01-11 | 亚星国际公司 | Symmetrical or semisymmetrical compounds useful as immunomodulators |
| WO2018119236A1 (en) * | 2016-12-22 | 2018-06-28 | Incyte Corporation | Triazolo[1,5-a]pyridine derivatives as immunomodulators |
Also Published As
| Publication number | Publication date |
|---|---|
| CN109897036A (en) | 2019-06-18 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN110128415B (en) | Indoline compounds used as immunomodulators and preparation method thereof | |
| CN109311851B (en) | Dihydro pyrimidinyl benzazepine carboxamide compounds | |
| CN109776377B (en) | Indoline compounds and preparation method and application thereof | |
| CN113444069B (en) | 2-aryl-4- (1H-pyrazol-3-yl) pyridine LSD1/HDAC double-target inhibitor | |
| CN105246887B (en) | Coumarin derivative and the method for treating hyperproliferative disease | |
| WO2016112846A1 (en) | 3-acetylenyl-pyrazole-pyrimidine derivative, and preparation method therefor and uses thereof | |
| WO2013013614A1 (en) | 4-(3-heteroarylarylamino)quinazoline and 1-(3-heteroarylarylamino)isoquinoline as hedgehog pathway inhibitor and use thereof | |
| CN110240629A (en) | Protein degradation targeting BCR-ABL compounds and their antitumor applications | |
| CN112442033A (en) | STING pathway modulators and uses thereof | |
| CN104211708B (en) | Benzoxazinone derivatives and its application as antiseptic | |
| CN114524799B (en) | HDAC inhibitor and preparation method and application thereof | |
| CN109897036B (en) | Triazolopyridine compounds and preparation method and use thereof | |
| TW479058B (en) | 2,7-substituted octahydro-pyrrolo[1,2-a]pyrazine derivatives | |
| CN114667289A (en) | Heteroaryl plasma kallikrein inhibitors | |
| CN105503627A (en) | Novel gossypol Schiff-base derivative and preparation and application thereof | |
| CN112876458B (en) | Apoptosis-ligand-1 antagonist compounds | |
| CN109280032A (en) | A kind of histone deacetylase inhibitor with pyridazinone core structure and preparation method and use thereof | |
| CN116178281B (en) | A kind of bifunctional immunosuppressant and its preparation method and application | |
| CN102898502A (en) | Coumarin derivative, and its preparation method and application | |
| CN110049973B (en) | 1,4-diphenyl-1H-imidazole and 2,4-diphenylthiazole derivatives and their preparation method and use | |
| CN116535395B (en) | 2H-indazole-7-carboxamide compounds, preparation method, pharmaceutical composition and application | |
| TW201922690A (en) | Inhibitors of cyclic-AMP response element-binding protein | |
| WO2015014283A1 (en) | Protein tyrosine kinase inhibitor and application thereof | |
| WO2024138292A1 (en) | Novel phenylquinolone compound having antibacterial and anticancer functions, and preparation thereof | |
| WO2018227886A1 (en) | New type indoleamine 2,3-dioxygenase inhibitor |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |