CN116925084A - 对肿瘤过表达的细胞周期调节蛋白wee1有双重抑制作用的化合物 - Google Patents
对肿瘤过表达的细胞周期调节蛋白wee1有双重抑制作用的化合物 Download PDFInfo
- Publication number
- CN116925084A CN116925084A CN202310470542.4A CN202310470542A CN116925084A CN 116925084 A CN116925084 A CN 116925084A CN 202310470542 A CN202310470542 A CN 202310470542A CN 116925084 A CN116925084 A CN 116925084A
- Authority
- CN
- China
- Prior art keywords
- htt
- compound
- wee1
- tumor
- pharmaceutically acceptable
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 150000001875 compounds Chemical class 0.000 title claims abstract description 149
- 101000621390 Homo sapiens Wee1-like protein kinase Proteins 0.000 title claims abstract description 95
- 102100023037 Wee1-like protein kinase Human genes 0.000 title claims abstract description 95
- 206010028980 Neoplasm Diseases 0.000 title claims abstract description 35
- 101710150974 Regulator of chromosome condensation Proteins 0.000 title abstract description 4
- 102100039977 Regulator of chromosome condensation Human genes 0.000 title abstract description 4
- 230000000694 effects Effects 0.000 title description 35
- 230000005764 inhibitory process Effects 0.000 title description 12
- 230000002401 inhibitory effect Effects 0.000 claims description 35
- 101710113864 Heat shock protein 90 Proteins 0.000 claims description 24
- 102100034051 Heat shock protein HSP 90-alpha Human genes 0.000 claims description 24
- 239000003814 drug Substances 0.000 claims description 22
- BKWJAKQVGHWELA-UHFFFAOYSA-N 1-[6-(2-hydroxypropan-2-yl)-2-pyridinyl]-6-[4-(4-methyl-1-piperazinyl)anilino]-2-prop-2-enyl-3-pyrazolo[3,4-d]pyrimidinone Chemical compound C1CN(C)CCN1C(C=C1)=CC=C1NC1=NC=C2C(=O)N(CC=C)N(C=3N=C(C=CC=3)C(C)(C)O)C2=N1 BKWJAKQVGHWELA-UHFFFAOYSA-N 0.000 claims description 18
- 229950009557 adavosertib Drugs 0.000 claims description 18
- 229940002612 prodrug Drugs 0.000 claims description 17
- 239000000651 prodrug Substances 0.000 claims description 17
- 150000003839 salts Chemical class 0.000 claims description 15
- 239000012453 solvate Substances 0.000 claims description 15
- 239000000126 substance Substances 0.000 claims description 15
- 229940079593 drug Drugs 0.000 claims description 13
- SWDZPNJZKUGIIH-QQTULTPQSA-N (5z)-n-ethyl-5-(4-hydroxy-6-oxo-3-propan-2-ylcyclohexa-2,4-dien-1-ylidene)-4-[4-(morpholin-4-ylmethyl)phenyl]-2h-1,2-oxazole-3-carboxamide Chemical compound O1NC(C(=O)NCC)=C(C=2C=CC(CN3CCOCC3)=CC=2)\C1=C1/C=C(C(C)C)C(O)=CC1=O SWDZPNJZKUGIIH-QQTULTPQSA-N 0.000 claims description 9
- 229950005069 luminespib Drugs 0.000 claims description 9
- 239000008194 pharmaceutical composition Substances 0.000 claims description 8
- 230000008685 targeting Effects 0.000 claims description 8
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 claims description 5
- 229940124597 therapeutic agent Drugs 0.000 claims description 5
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 4
- 239000002246 antineoplastic agent Substances 0.000 claims description 4
- 125000002915 carbonyl group Chemical group [*:2]C([*:1])=O 0.000 claims description 4
- 239000003795 chemical substances by application Substances 0.000 claims description 4
- 125000004435 hydrogen atom Chemical group [H]* 0.000 claims description 4
- 125000005647 linker group Chemical group 0.000 claims description 4
- 238000002360 preparation method Methods 0.000 claims description 3
- 125000003161 (C1-C6) alkylene group Chemical group 0.000 claims description 2
- BWGNESOTFCXPMA-UHFFFAOYSA-N Dihydrogen disulfide Chemical compound SS BWGNESOTFCXPMA-UHFFFAOYSA-N 0.000 claims description 2
- 125000002947 alkylene group Chemical group 0.000 claims description 2
- 150000001408 amides Chemical class 0.000 claims description 2
- 229940127089 cytotoxic agent Drugs 0.000 claims description 2
- 239000003937 drug carrier Substances 0.000 claims description 2
- 239000002955 immunomodulating agent Substances 0.000 claims description 2
- 230000002265 prevention Effects 0.000 claims description 2
- 125000000896 monocarboxylic acid group Chemical group 0.000 claims 2
- 239000003112 inhibitor Substances 0.000 abstract description 39
- 239000003481 heat shock protein 90 inhibitor Substances 0.000 abstract description 19
- 230000000259 anti-tumor effect Effects 0.000 abstract description 12
- 230000007547 defect Effects 0.000 abstract description 6
- 231100000086 high toxicity Toxicity 0.000 abstract description 2
- 229940043355 kinase inhibitor Drugs 0.000 abstract description 2
- 239000003757 phosphotransferase inhibitor Substances 0.000 abstract description 2
- 239000007787 solid Substances 0.000 description 35
- 238000006243 chemical reaction Methods 0.000 description 28
- 230000015572 biosynthetic process Effects 0.000 description 26
- 238000003786 synthesis reaction Methods 0.000 description 25
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 21
- 210000004027 cell Anatomy 0.000 description 21
- 238000002474 experimental method Methods 0.000 description 17
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 16
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 15
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 15
- 230000015556 catabolic process Effects 0.000 description 15
- 239000012043 crude product Substances 0.000 description 15
- 238000006731 degradation reaction Methods 0.000 description 15
- JGFZNNIVVJXRND-UHFFFAOYSA-N N,N-Diisopropylethylamine (DIPEA) Chemical compound CCN(C(C)C)C(C)C JGFZNNIVVJXRND-UHFFFAOYSA-N 0.000 description 14
- 239000000047 product Substances 0.000 description 14
- 210000004881 tumor cell Anatomy 0.000 description 14
- 102000004169 proteins and genes Human genes 0.000 description 13
- 108090000623 proteins and genes Proteins 0.000 description 13
- 230000000144 pharmacologic effect Effects 0.000 description 12
- 108091008611 Protein Kinase B Proteins 0.000 description 11
- 102100033810 RAC-alpha serine/threonine-protein kinase Human genes 0.000 description 11
- 230000002829 reductive effect Effects 0.000 description 10
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 9
- 230000008684 selective degradation Effects 0.000 description 9
- 239000000243 solution Substances 0.000 description 9
- HEDRZPFGACZZDS-MICDWDOJSA-N Trichloro(2H)methane Chemical compound [2H]C(Cl)(Cl)Cl HEDRZPFGACZZDS-MICDWDOJSA-N 0.000 description 8
- 238000001514 detection method Methods 0.000 description 8
- 229910052757 nitrogen Inorganic materials 0.000 description 8
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 8
- RVAQIUULWULRNW-UHFFFAOYSA-N Ganetespib Chemical compound C1=C(O)C(C(C)C)=CC(C=2N(C(O)=NN=2)C=2C=C3C=CN(C)C3=CC=2)=C1O RVAQIUULWULRNW-UHFFFAOYSA-N 0.000 description 7
- 238000002347 injection Methods 0.000 description 7
- 239000007924 injection Substances 0.000 description 7
- 238000010791 quenching Methods 0.000 description 7
- 230000000171 quenching effect Effects 0.000 description 7
- 239000013543 active substance Substances 0.000 description 6
- 238000001035 drying Methods 0.000 description 6
- 238000010898 silica gel chromatography Methods 0.000 description 6
- 238000001308 synthesis method Methods 0.000 description 6
- 238000010189 synthetic method Methods 0.000 description 6
- DTQVDTLACAAQTR-UHFFFAOYSA-N trifluoroacetic acid Substances OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 description 6
- 230000009471 action Effects 0.000 description 5
- 201000011510 cancer Diseases 0.000 description 5
- 230000001404 mediated effect Effects 0.000 description 5
- 238000000034 method Methods 0.000 description 5
- 238000011865 proteolysis targeting chimera technique Methods 0.000 description 5
- 238000011160 research Methods 0.000 description 5
- 108010026668 snake venom protein C activator Proteins 0.000 description 5
- 108020004414 DNA Proteins 0.000 description 4
- 239000007821 HATU Substances 0.000 description 4
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 4
- 230000001028 anti-proliverative effect Effects 0.000 description 4
- 230000003828 downregulation Effects 0.000 description 4
- 238000000338 in vitro Methods 0.000 description 4
- 239000003446 ligand Substances 0.000 description 4
- 230000004048 modification Effects 0.000 description 4
- 238000012986 modification Methods 0.000 description 4
- 230000037361 pathway Effects 0.000 description 4
- 230000035755 proliferation Effects 0.000 description 4
- 239000002904 solvent Substances 0.000 description 4
- 210000001519 tissue Anatomy 0.000 description 4
- 206010006187 Breast cancer Diseases 0.000 description 3
- 208000026310 Breast neoplasm Diseases 0.000 description 3
- 108050001186 Chaperonin Cpn60 Proteins 0.000 description 3
- 102000052603 Chaperonins Human genes 0.000 description 3
- 206010061535 Ovarian neoplasm Diseases 0.000 description 3
- KDLHZDBZIXYQEI-UHFFFAOYSA-N Palladium Chemical compound [Pd] KDLHZDBZIXYQEI-UHFFFAOYSA-N 0.000 description 3
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 description 3
- 230000004071 biological effect Effects 0.000 description 3
- 229940125782 compound 2 Drugs 0.000 description 3
- 239000002552 dosage form Substances 0.000 description 3
- 208000032839 leukemia Diseases 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 239000013641 positive control Substances 0.000 description 3
- 239000002994 raw material Substances 0.000 description 3
- 238000010998 test method Methods 0.000 description 3
- 231100000331 toxic Toxicity 0.000 description 3
- 230000002588 toxic effect Effects 0.000 description 3
- 238000013518 transcription Methods 0.000 description 3
- 230000035897 transcription Effects 0.000 description 3
- 238000012546 transfer Methods 0.000 description 3
- 238000011144 upstream manufacturing Methods 0.000 description 3
- VUDZSIYXZUYWSC-DBRKOABJSA-N (4r)-1-[(2r,4r,5r)-3,3-difluoro-4-hydroxy-5-(hydroxymethyl)oxolan-2-yl]-4-hydroxy-1,3-diazinan-2-one Chemical compound FC1(F)[C@H](O)[C@@H](CO)O[C@H]1N1C(=O)N[C@H](O)CC1 VUDZSIYXZUYWSC-DBRKOABJSA-N 0.000 description 2
- OXTSYWDBUVRXFF-GDLZYMKVSA-N 1-[(7R)-7-ethyl-7-hydroxy-5,6-dihydrocyclopenta[b]pyridin-2-yl]-6-[4-(4-methylpiperazin-1-yl)anilino]-2-prop-2-enylpyrazolo[3,4-d]pyrimidin-3-one Chemical compound C1=C2C(=NC(N3C4=NC(NC5=CC=C(C=C5)N5CCN(CC5)C)=NC=C4C(=O)N3CC=C)=C1)[C@@](CC)(CC2)O OXTSYWDBUVRXFF-GDLZYMKVSA-N 0.000 description 2
- MWDVCHRYCKXEBY-LBPRGKRZSA-N 3-chloro-n-[2-oxo-2-[[(1s)-1-phenylethyl]amino]ethyl]benzamide Chemical compound N([C@@H](C)C=1C=CC=CC=1)C(=O)CNC(=O)C1=CC=CC(Cl)=C1 MWDVCHRYCKXEBY-LBPRGKRZSA-N 0.000 description 2
- JOFDSYLCZIHGGO-UHFFFAOYSA-N 4-[(4-cyclohexylphenyl)methyl-[2-[[5-(dimethylamino)naphthalen-1-yl]sulfonyl-methylamino]acetyl]amino]-2-hydroxybenzoic acid Chemical compound C1=CC=C2C(N(C)C)=CC=CC2=C1S(=O)(=O)N(C)CC(=O)N(C=1C=C(O)C(C(O)=O)=CC=1)CC(C=C1)=CC=C1C1CCCCC1 JOFDSYLCZIHGGO-UHFFFAOYSA-N 0.000 description 2
- 102000007469 Actins Human genes 0.000 description 2
- 108010085238 Actins Proteins 0.000 description 2
- 102100025064 Cellular tumor antigen p53 Human genes 0.000 description 2
- 206010008342 Cervix carcinoma Diseases 0.000 description 2
- 206010009944 Colon cancer Diseases 0.000 description 2
- 102100032857 Cyclin-dependent kinase 1 Human genes 0.000 description 2
- 101710106279 Cyclin-dependent kinase 1 Proteins 0.000 description 2
- 108010006519 Molecular Chaperones Proteins 0.000 description 2
- 102000005431 Molecular Chaperones Human genes 0.000 description 2
- 206010033128 Ovarian cancer Diseases 0.000 description 2
- 208000005718 Stomach Neoplasms Diseases 0.000 description 2
- 239000006180 TBST buffer Substances 0.000 description 2
- 208000006105 Uterine Cervical Neoplasms Diseases 0.000 description 2
- 239000000611 antibody drug conjugate Substances 0.000 description 2
- 229940049595 antibody-drug conjugate Drugs 0.000 description 2
- 238000003556 assay Methods 0.000 description 2
- 230000000903 blocking effect Effects 0.000 description 2
- 230000001413 cellular effect Effects 0.000 description 2
- 201000010881 cervical cancer Diseases 0.000 description 2
- 229940044683 chemotherapy drug Drugs 0.000 description 2
- 239000000562 conjugate Substances 0.000 description 2
- 239000008380 degradant Substances 0.000 description 2
- 238000004090 dissolution Methods 0.000 description 2
- 206010017758 gastric cancer Diseases 0.000 description 2
- 229910052739 hydrogen Inorganic materials 0.000 description 2
- 239000001257 hydrogen Substances 0.000 description 2
- 201000007270 liver cancer Diseases 0.000 description 2
- 208000014018 liver neoplasm Diseases 0.000 description 2
- 230000007246 mechanism Effects 0.000 description 2
- 230000010534 mechanism of action Effects 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 230000004060 metabolic process Effects 0.000 description 2
- 230000011278 mitosis Effects 0.000 description 2
- HBEDNENASUYMPO-LJQANCHMSA-N n-hydroxy-4-[[(2r)-3-oxo-2-(thiophen-2-ylmethyl)-2,4-dihydroquinoxalin-1-yl]methyl]benzamide Chemical compound C1=CC(C(=O)NO)=CC=C1CN1C2=CC=CC=C2NC(=O)[C@H]1CC1=CC=CS1 HBEDNENASUYMPO-LJQANCHMSA-N 0.000 description 2
- 230000000865 phosphorylative effect Effects 0.000 description 2
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 description 2
- 229940124823 proteolysis targeting chimeric molecule Drugs 0.000 description 2
- 230000008439 repair process Effects 0.000 description 2
- 239000000741 silica gel Substances 0.000 description 2
- 229910002027 silica gel Inorganic materials 0.000 description 2
- 229940126586 small molecule drug Drugs 0.000 description 2
- 239000007858 starting material Substances 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 201000011549 stomach cancer Diseases 0.000 description 2
- 230000001225 therapeutic effect Effects 0.000 description 2
- GHYOCDFICYLMRF-UTIIJYGPSA-N (2S,3R)-N-[(2S)-3-(cyclopenten-1-yl)-1-[(2R)-2-methyloxiran-2-yl]-1-oxopropan-2-yl]-3-hydroxy-3-(4-methoxyphenyl)-2-[[(2S)-2-[(2-morpholin-4-ylacetyl)amino]propanoyl]amino]propanamide Chemical compound C1(=CCCC1)C[C@@H](C(=O)[C@@]1(OC1)C)NC([C@H]([C@@H](C1=CC=C(C=C1)OC)O)NC([C@H](C)NC(CN1CCOCC1)=O)=O)=O GHYOCDFICYLMRF-UTIIJYGPSA-N 0.000 description 1
- QFLWZFQWSBQYPS-AWRAUJHKSA-N (3S)-3-[[(2S)-2-[[(2S)-2-[5-[(3aS,6aR)-2-oxo-1,3,3a,4,6,6a-hexahydrothieno[3,4-d]imidazol-4-yl]pentanoylamino]-3-methylbutanoyl]amino]-3-(4-hydroxyphenyl)propanoyl]amino]-4-[1-bis(4-chlorophenoxy)phosphorylbutylamino]-4-oxobutanoic acid Chemical compound CCCC(NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](Cc1ccc(O)cc1)NC(=O)[C@@H](NC(=O)CCCCC1SC[C@@H]2NC(=O)N[C@H]12)C(C)C)P(=O)(Oc1ccc(Cl)cc1)Oc1ccc(Cl)cc1 QFLWZFQWSBQYPS-AWRAUJHKSA-N 0.000 description 1
- FALRKNHUBBKYCC-UHFFFAOYSA-N 2-(chloromethyl)pyridine-3-carbonitrile Chemical compound ClCC1=NC=CC=C1C#N FALRKNHUBBKYCC-UHFFFAOYSA-N 0.000 description 1
- UOXJNGFFPMOZDM-UHFFFAOYSA-N 2-[di(propan-2-yl)amino]ethylsulfanyl-methylphosphinic acid Chemical compound CC(C)N(C(C)C)CCSP(C)(O)=O UOXJNGFFPMOZDM-UHFFFAOYSA-N 0.000 description 1
- SFHYNDMGZXWXBU-LIMNOBDPSA-N 6-amino-2-[[(e)-(3-formylphenyl)methylideneamino]carbamoylamino]-1,3-dioxobenzo[de]isoquinoline-5,8-disulfonic acid Chemical compound O=C1C(C2=3)=CC(S(O)(=O)=O)=CC=3C(N)=C(S(O)(=O)=O)C=C2C(=O)N1NC(=O)N\N=C\C1=CC=CC(C=O)=C1 SFHYNDMGZXWXBU-LIMNOBDPSA-N 0.000 description 1
- 208000010507 Adenocarcinoma of Lung Diseases 0.000 description 1
- 206010004593 Bile duct cancer Diseases 0.000 description 1
- 206010005003 Bladder cancer Diseases 0.000 description 1
- 208000003174 Brain Neoplasms Diseases 0.000 description 1
- 101100520513 Caenorhabditis elegans wee-1.3 gene Proteins 0.000 description 1
- 201000009030 Carcinoma Diseases 0.000 description 1
- 102400000888 Cholecystokinin-8 Human genes 0.000 description 1
- 101800005151 Cholecystokinin-8 Proteins 0.000 description 1
- 108010077544 Chromatin Proteins 0.000 description 1
- 208000001333 Colorectal Neoplasms Diseases 0.000 description 1
- 102000003910 Cyclin D Human genes 0.000 description 1
- 108090000259 Cyclin D Proteins 0.000 description 1
- 102000003909 Cyclin E Human genes 0.000 description 1
- 108090000257 Cyclin E Proteins 0.000 description 1
- 108090000266 Cyclin-dependent kinases Proteins 0.000 description 1
- 102000003903 Cyclin-dependent kinases Human genes 0.000 description 1
- 230000005971 DNA damage repair Effects 0.000 description 1
- 230000033616 DNA repair Effects 0.000 description 1
- 102000004163 DNA-directed RNA polymerases Human genes 0.000 description 1
- 108090000626 DNA-directed RNA polymerases Proteins 0.000 description 1
- 102100030013 Endoribonuclease Human genes 0.000 description 1
- 101710199605 Endoribonuclease Proteins 0.000 description 1
- 208000000461 Esophageal Neoplasms Diseases 0.000 description 1
- GHASVSINZRGABV-UHFFFAOYSA-N Fluorouracil Chemical compound FC1=CNC(=O)NC1=O GHASVSINZRGABV-UHFFFAOYSA-N 0.000 description 1
- 230000008051 G1/S transition checkpoint Effects 0.000 description 1
- 230000004668 G2/M phase Effects 0.000 description 1
- 206010059024 Gastrointestinal toxicity Diseases 0.000 description 1
- 208000032612 Glial tumor Diseases 0.000 description 1
- 206010018338 Glioma Diseases 0.000 description 1
- 102100031181 Glyceraldehyde-3-phosphate dehydrogenase Human genes 0.000 description 1
- 101710103773 Histone H2B Proteins 0.000 description 1
- 102100021639 Histone H2B type 1-K Human genes 0.000 description 1
- 101000603402 Homo sapiens Protein NPAT Proteins 0.000 description 1
- 101000621401 Homo sapiens Wee1-like protein kinase 2 Proteins 0.000 description 1
- 108010001336 Horseradish Peroxidase Proteins 0.000 description 1
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 1
- 208000008839 Kidney Neoplasms Diseases 0.000 description 1
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 description 1
- 206010058467 Lung neoplasm malignant Diseases 0.000 description 1
- 206010025323 Lymphomas Diseases 0.000 description 1
- BZLVMXJERCGZMT-UHFFFAOYSA-N Methyl tert-butyl ether Chemical compound COC(C)(C)C BZLVMXJERCGZMT-UHFFFAOYSA-N 0.000 description 1
- 208000034578 Multiple myelomas Diseases 0.000 description 1
- 102000006386 Myelin Proteins Human genes 0.000 description 1
- 108010083674 Myelin Proteins Proteins 0.000 description 1
- 102100038893 Myelin transcription factor 1 Human genes 0.000 description 1
- 101710104371 Myelin transcription factor 1 Proteins 0.000 description 1
- 206010028813 Nausea Diseases 0.000 description 1
- 206010029350 Neurotoxicity Diseases 0.000 description 1
- 239000000020 Nitrocellulose Substances 0.000 description 1
- 206010030155 Oesophageal carcinoma Diseases 0.000 description 1
- 108700020796 Oncogene Proteins 0.000 description 1
- 206010061902 Pancreatic neoplasm Diseases 0.000 description 1
- 108091000080 Phosphotransferase Proteins 0.000 description 1
- 206010035226 Plasma cell myeloma Diseases 0.000 description 1
- 206010060862 Prostate cancer Diseases 0.000 description 1
- 208000000236 Prostatic Neoplasms Diseases 0.000 description 1
- 102100038870 Protein NPAT Human genes 0.000 description 1
- 206010038389 Renal cancer Diseases 0.000 description 1
- 230000018199 S phase Effects 0.000 description 1
- 101710113029 Serine/threonine-protein kinase Proteins 0.000 description 1
- 208000000453 Skin Neoplasms Diseases 0.000 description 1
- 208000024313 Testicular Neoplasms Diseases 0.000 description 1
- 206010057644 Testis cancer Diseases 0.000 description 1
- 206010044221 Toxic encephalopathy Diseases 0.000 description 1
- 108091023040 Transcription factor Proteins 0.000 description 1
- 102000040945 Transcription factor Human genes 0.000 description 1
- 102000006275 Ubiquitin-Protein Ligases Human genes 0.000 description 1
- 108010083111 Ubiquitin-Protein Ligases Proteins 0.000 description 1
- 208000007097 Urinary Bladder Neoplasms Diseases 0.000 description 1
- 206010046798 Uterine leiomyoma Diseases 0.000 description 1
- 102100023040 Wee1-like protein kinase 2 Human genes 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 239000012190 activator Substances 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 239000002168 alkylating agent Substances 0.000 description 1
- 229940100198 alkylating agent Drugs 0.000 description 1
- 125000003277 amino group Chemical group 0.000 description 1
- 230000003698 anagen phase Effects 0.000 description 1
- 208000007502 anemia Diseases 0.000 description 1
- 230000001093 anti-cancer Effects 0.000 description 1
- 229940041181 antineoplastic drug Drugs 0.000 description 1
- 230000006907 apoptotic process Effects 0.000 description 1
- 208000026900 bile duct neoplasm Diseases 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- 210000000481 breast Anatomy 0.000 description 1
- 210000004899 c-terminal region Anatomy 0.000 description 1
- VSJKWCGYPAHWDS-FQEVSTJZSA-N camptothecin Chemical compound C1=CC=C2C=C(CN3C4=CC5=C(C3=O)COC(=O)[C@]5(O)CC)C4=NC2=C1 VSJKWCGYPAHWDS-FQEVSTJZSA-N 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 239000011203 carbon fibre reinforced carbon Substances 0.000 description 1
- 238000006555 catalytic reaction Methods 0.000 description 1
- 230000032823 cell division Effects 0.000 description 1
- 239000013592 cell lysate Substances 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 125000003636 chemical group Chemical group 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 208000006990 cholangiocarcinoma Diseases 0.000 description 1
- 210000003483 chromatin Anatomy 0.000 description 1
- 208000029742 colonic neoplasm Diseases 0.000 description 1
- 238000004440 column chromatography Methods 0.000 description 1
- 229940125797 compound 12 Drugs 0.000 description 1
- 229940125898 compound 5 Drugs 0.000 description 1
- 230000002508 compound effect Effects 0.000 description 1
- 230000002950 deficient Effects 0.000 description 1
- 230000030609 dephosphorylation Effects 0.000 description 1
- 238000006209 dephosphorylation reaction Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 231100000371 dose-limiting toxicity Toxicity 0.000 description 1
- 230000005782 double-strand break Effects 0.000 description 1
- 230000002222 downregulating effect Effects 0.000 description 1
- 230000001516 effect on protein Effects 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 210000002919 epithelial cell Anatomy 0.000 description 1
- 201000004101 esophageal cancer Diseases 0.000 description 1
- 230000005496 eutectics Effects 0.000 description 1
- 230000029142 excretion Effects 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
- 229960002949 fluorouracil Drugs 0.000 description 1
- 231100000414 gastrointestinal toxicity Toxicity 0.000 description 1
- SDUQYLNIPVEERB-QPPQHZFASA-N gemcitabine Chemical compound O=C1N=C(N)C=CN1[C@H]1C(F)(F)[C@H](O)[C@@H](CO)O1 SDUQYLNIPVEERB-QPPQHZFASA-N 0.000 description 1
- 229960005277 gemcitabine Drugs 0.000 description 1
- 208000005017 glioblastoma Diseases 0.000 description 1
- 108020004445 glyceraldehyde-3-phosphate dehydrogenase Proteins 0.000 description 1
- 230000009036 growth inhibition Effects 0.000 description 1
- 208000024348 heart neoplasm Diseases 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 231100000304 hepatotoxicity Toxicity 0.000 description 1
- 150000002431 hydrogen Chemical class 0.000 description 1
- 230000001024 immunotherapeutic effect Effects 0.000 description 1
- 230000002779 inactivation Effects 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 201000010982 kidney cancer Diseases 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 201000010260 leiomyoma Diseases 0.000 description 1
- 230000003902 lesion Effects 0.000 description 1
- 230000001665 lethal effect Effects 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- -1 linker compound Chemical class 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 230000007056 liver toxicity Effects 0.000 description 1
- 239000012160 loading buffer Substances 0.000 description 1
- 231100000053 low toxicity Toxicity 0.000 description 1
- 201000005249 lung adenocarcinoma Diseases 0.000 description 1
- 201000005202 lung cancer Diseases 0.000 description 1
- 208000020816 lung neoplasm Diseases 0.000 description 1
- 208000015486 malignant pancreatic neoplasm Diseases 0.000 description 1
- 230000035800 maturation Effects 0.000 description 1
- 201000001441 melanoma Diseases 0.000 description 1
- 230000001394 metastastic effect Effects 0.000 description 1
- 206010061289 metastatic neoplasm Diseases 0.000 description 1
- UKVIEHSSVKSQBA-UHFFFAOYSA-N methane;palladium Chemical compound C.[Pd] UKVIEHSSVKSQBA-UHFFFAOYSA-N 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- 210000005012 myelin Anatomy 0.000 description 1
- 230000008693 nausea Effects 0.000 description 1
- 239000013642 negative control Substances 0.000 description 1
- 231100000228 neurotoxicity Toxicity 0.000 description 1
- 230000007135 neurotoxicity Effects 0.000 description 1
- 229920001220 nitrocellulos Polymers 0.000 description 1
- 208000002154 non-small cell lung carcinoma Diseases 0.000 description 1
- 230000009437 off-target effect Effects 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 201000002528 pancreatic cancer Diseases 0.000 description 1
- 208000008443 pancreatic carcinoma Diseases 0.000 description 1
- 102000020233 phosphotransferase Human genes 0.000 description 1
- 229910052697 platinum Inorganic materials 0.000 description 1
- 231100000683 possible toxicity Toxicity 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000017854 proteolysis Effects 0.000 description 1
- 230000005855 radiation Effects 0.000 description 1
- 239000011541 reaction mixture Substances 0.000 description 1
- 238000010992 reflux Methods 0.000 description 1
- IZTQOLKUZKXIRV-YRVFCXMDSA-N sincalide Chemical compound C([C@@H](C(=O)N[C@@H](CCSC)C(=O)NCC(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC=1C=CC=CC=1)C(N)=O)NC(=O)[C@@H](N)CC(O)=O)C1=CC=C(OS(O)(=O)=O)C=C1 IZTQOLKUZKXIRV-YRVFCXMDSA-N 0.000 description 1
- 230000005783 single-strand break Effects 0.000 description 1
- 201000000849 skin cancer Diseases 0.000 description 1
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 1
- 239000012265 solid product Substances 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 229940014800 succinic anhydride Drugs 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 229920002994 synthetic fiber Polymers 0.000 description 1
- 239000003826 tablet Substances 0.000 description 1
- 201000003120 testicular cancer Diseases 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 229940126585 therapeutic drug Drugs 0.000 description 1
- 206010043554 thrombocytopenia Diseases 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 208000029729 tumor suppressor gene on chromosome 11 Diseases 0.000 description 1
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 description 1
- 230000034512 ubiquitination Effects 0.000 description 1
- 238000010798 ubiquitination Methods 0.000 description 1
- 201000005112 urinary bladder cancer Diseases 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 238000001262 western blot Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
- C07D487/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains two hetero rings
- C07D487/04—Ortho-condensed systems
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Pharmacology & Pharmacy (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
本发明涉及细胞周期调节蛋白WEE1激酶抑制剂与HSP90抑制剂形成的化合物,其具有良好的抗肿瘤活性,可克服一般WEE1抑制剂对肿瘤组织选择性差,毒性高等缺点。
Description
技术领域
本发明涉及WEE1激酶抑制剂与HSP90抑制剂形成的化合物,其可达到双重WEE1抑制作用,具有良好的抗肿瘤活性,可克服一般WEE1抑制剂抗肿瘤靶向性差,毒性高的缺点。
背景技术
WEE1激酶是丝氨酸/苏氨酸蛋白激酶家族的重要成员之一,是一种高度保守的细胞周期调节蛋白。人类WEE家族包括髓鞘转录因子1(myelin transcription factor 1,Myt1)、WEE1、WEE2。WEE1主要包含3个结构域:N-端结构域、中心激酶结构域和1个短的C-端结构域,其中最重要的N端结构域是WEE1激酶的激活结构域,也是抑制周期蛋白依赖性激酶1-周期蛋白B(CDK1-CyclinB)复合物去磷酸化的潜在位点,这些泛素化识别位点可以严格控制WEE1在细胞内的半衰期。
当细胞受烷化剂或辐射等刺激后,导致DNA发生单链或双链断裂。WEE1激酶可通过2条途径阻滞细胞的有丝***:一是通过磷酸化组蛋白H2B,抑制转录激活因子NPAT和RNA聚合酶结合,下调H2B表达,对细胞***的S期产生阻滞;二是通过磷酸化CDK1的第15位酪氨酸,抑制CDK1活性,产生G2/M阻滞,为修复受损DNA争取时间。待DNA修复完成后再进入有丝***,从而维持DNA和染色质的完整性。
在正常细胞中,DNA损伤修复不仅可以通过WEE1激酶介导的两条途径,还可通过抑癌基因P53介导的2种通路:ATM/ATR-P53-CDK4/Cyclin D和ATM/ATR-P53-CDK2/Cyclin E。然而,大部分的肿瘤细胞会发生P53突变并导致G1/S检查点失活,使得WEE1介导的G2/M修复成为DNA的主要依赖方式。WEE1抑制剂能够使细胞未经修复直接进入***期,从而产生合成致死效应。理论上,抑制WEE1激酶活性能够选择性地杀死p53功能缺陷的肿瘤细胞,而不杀死正常细胞。相关研究表明,WEE1激酶在肝癌、肺癌、胃癌、乳腺癌、白血病、***、胶质母细胞瘤等癌症中过表达。因此,抑制或降解WEE1成为一种理想的肿瘤治疗途径。
目前已报道多个WEE1抑制剂,但无一被批准上市。该靶点目前研究进展最快的候选药物是AZD1775和ZN-C3,目前均处于临床二期。ZN-C3是近年来异军突起的一种WEE1抑制剂,相关研究并不多,且目前临床数据披露较少,其疗效和潜在毒性尚不明确。AZD1775与靶点结合的共晶结构已经阐明,其在临床研究的主要适应症为肿瘤相关疾病,包括子宫肌瘤、白血病、乳腺癌和卵巢癌等。目前,AZD1775在临床二期中对卵巢癌治疗效果最佳,进一步的研究还在有序开展中。AZD1775既可以单独用药,也可以与吉西他滨、铂类、5-氟尿嘧啶以及喜树碱等化疗药物联合使用。尽管AZD1775等WEE1抑制剂在临床上显示出较好的疗效,但其普遍存在剂量限制性毒性和脱靶效应,导致血小板减少、贫血和恶心等副作用。这些缺陷阻碍了WEE1抑制剂的进一步发展,可通过开发高选择性的WEE1抑制剂来改善现状。
近年来,针对WEE1的靶向降解剂PROTACs也进入了公众视野,其中化合物ZNL-02-096在诱导细胞凋亡和G2/M期积聚的剂量比AZD1775低10倍,证明WEE1降解剂可以在较低剂量下发挥出比WEE1抑制剂更强的功效。WEE1的降解剂能够以PROTAC的独特药物作用机制在催化量下发挥抗肿瘤作用,并且有望克服WEE1抑制剂的许多不足,但至今仍无任何一款WEE1降解剂进入临床,可能的原因是UPS***中E3连接酶无组织特异性,导致药物的选择性差。
为了克服针对WEE1靶点的抑制剂和PROTAC的缺点,提高药物的抗肿瘤活性降低毒副作用。一方面,可以开发具有高选择性、低毒性的WEE1抑制剂;另一方面,可以通过探索WEE1抑制剂与其他药物联用或制成偶联化合物的方式来提高药物活性。
HSP90作为一种伴侣蛋白,是细胞内含量最高的蛋白之一,约占细胞蛋白总量的1-3%,本身也是一种已知的肿瘤靶点。由于HSP90分布较广,且通常在肿瘤组织中高表达,因此针对该靶点的抑制剂具有较强的肿瘤组织选择性,HSP90近年来逐步成为抗肿瘤药物研究的热点。临床实验结果表明,HSP90抑制剂往往需要高剂量才能起效。正因为如此,HSP90在临床表现出较多的副作用,例如视觉毒性、胃肠道毒性、肝毒性、神经毒性等,大大的限制了其应用,至今也还未有HSP90抑制剂被批准上市。另外,本领域虽有将HSP90抑制剂与其它抗癌物质组合应用的报道,但尚无将HSP90抑制剂与WEE1抑制剂进行组合、特别是制成化合物的报道。
发明内容
本发明在实验中发现,由于HSP90在肿瘤细胞中的持续分泌,因此当WEE1抑制剂与HSP90抑制剂形成化合物时,可利用HSP90抑制剂对肿瘤细胞进行靶向,由此显著提高WEE1抑制剂在肿瘤细胞中抑制WEE1激酶的作用,进而解决现有WEE1抑制剂选择性,毒副作用大的缺点,使WEE1抑制剂上市应用成为可能。另外,本发明还发现HSP90作为伴侣蛋白可以介导四百多种客户蛋白的折叠和成熟。WEE1作为其客户蛋白之一,在本发明的化合物中或许能通过HSP90抑制剂的作用实现选择性降解。
因此,本发明涉及一种化合物,其包括WEE1抑制部分与HSP90抑制部分,两者通过连接链连接;或其药学上可接受的盐、溶剂合物、互变异构体、立体异构体、外消旋体或前药。
在一个实施方案中,前述连接链是指在HSP90抑制部分和WEE1抑制部分之间形成共价结合的键或者连接基团,这些连接链包括:化学键(包括碳碳键、碳氮键、碳硫键、碳磷键、氮氮键)、酯键(包括羧酸酯键、磺酸酯键、氨基甲酸酯键)、羰基(-C(=O)-)、C1-6亚烷基、酰胺键、醚键、二硫键等及其组合。
在所述化合物的一个优选实施方案中,HSP90抑制部分包括但不限于如下结构,以及由这些结构衍生而成的可以和连接链形成各类共价键的结构或基团:
或其药学上可接受的盐、溶剂合物、互变异构体、立体异构体、外消旋体或前药。
在所述化合物的一个优选实施方案中,WEE1抑制部分包括但不限于如下结构,以及由这些结构衍生而成的可以和连接链形成各类共价键的结构或基团:
或其药学上可接受的盐、溶剂合物、互变异构体、立体异构体、外消旋体或前药。
在所述化合物的一个优选的实施方案中,HSP90抑制部分包括上述NVP-AUY922、STA9090或AT13387类似物的结构,或由NVP-AUY922、STA9090或AT13387类似物衍生的结构;WEE1抑制部分包括上述AZD1775的结构,或由AZD1775衍生的结构;连接链选自化学键、酰胺键、羰基、C1-6亚烷基及其组合,
或其药学上可接受的盐、溶剂合物、互变异构体、立体异构体、外消旋体或前药。
在所述化合物的一个更优选的实施方案中,
HSP90抑制部分包含从以下结构中除去一个氢原子后得到的一价基团:
其中R1表示-COOH,R2表示-NHC(=O)CH2CH2COOH,R3表示-CH2COOH;
WEE1抑制部分包含从以下结构中除去一个氢原子后得到的一价基团:
连接链选自化学键:-NH-(CH2)n-、-NH-(CH2)2-Ph-O-C(=O)-,
或其药学上可接受的盐、溶剂合物、互变异构体、立体异构体、外消旋体或前药。
在一个进一步优选的实施方案中,本发明的化合物为如下所示的HTT-19~HTT-30,
或其药学上可接受的盐、溶剂合物、互变异构体、立体异构体、外消旋体或前药。
在一个实施方案中,本发明涉及一种药物组合物,其包含如前文所述的本发明的化合物或其药学上可接受的盐、溶剂合物、互变异构体、立体异构体、外消旋体或前药,以及一种或多种药用载体。
在一个实施方案中,本发明的药物组合物中还包含其它治疗药物。在另一个实施方案中,本发明的药物组合物与其它治疗药物组合使用。所述其它治疗药物例如为肿瘤化疗药物、肿瘤靶向药物、肿瘤免疫治疗药物、肿瘤药物偶联物(例如抗体药物偶联物、小分子药物化合物和微型药物化合物)。
另一个实施方案涉及本发明的化合物或药物组合物用于制备药物的用途,所述药物用于预防或***。
本发明的化合物具有良好的WEE1抑制活性和HSP90抑制活性,特别是由于HSP90抑制活性所带来的靶向作用而使得WEE1抑制活性得以增强,解决了WEE1抑制剂靶向性差、毒副作用大的缺点,为癌症的临床治疗提供了有希望的选择。如本说明书药理实施例部分所示,本发明的化合物在癌细胞株(MV-4-11)中,取得了显著的癌症细胞抑制效果。
附图说明
图1;图3;图4;图5;图6:化合物HTT-22~HTT-30的Western实验结果。
图2:化合物HTT-21对MV-4-11的抗增殖实验结果。
图7:化合物HTT-19~HTT-21对WEE1转录组的影响。
图8:WEE1抑制剂AZD1775对WEE1转录组的影响。
具体实施方式
本说明书所用术语“改造”是指出于将具有某种活性的物质(如WEE1抑制剂和HSP90抑制剂)制成化合物的目的,而将该化合物进行结构修饰,但修饰后的部分仍包含原物质的主体结构或活性结构,并保留(甚至在形成化合物后可能提高)原抑制剂的药理活性。
本说明书所用术语“抑制部分”(如WEE1抑制部分和HSP90抑制部分)是指将相应的抑制剂(如WEE1抑制剂和HSP90抑制剂)经过上述改造后得到的部分,该部分可为一价或更高价基团,并且与连接链连接。
本说明书所用术语“连接链”是指连接两个或多个抑制部分的化学部分,可为如前文所定义的化学键或化学基团。连接链(或其部分)可存在于改造前的活性物质分子或改造后的抑制部分中,也可为新增加的连接键或基团。
本说明书所用术语“化合物”是指通过连接链将至少两个上述抑制部分连接后而获得的化合物。
本说明书所用术语“前药”是以非活性或非完全活性形式给药并随后通过代谢过程转化成活性物质(例如本发明的化合物)的物质。前药可用于改进活性物质的吸收、分布、代谢和/或***,也可用于改进活性物质针对特定细胞或过程相互作用的选择性,由此例如可降低活性物质的不良反应或非预期作用。
本说明书所用术语“约”表示在相应数值上下浮动10%的范围。例如,若某种成分的浓度为约5mM,表明其浓度为4.5-5.5mM;若某种成分的浓度范围为约5-10mM,表明其浓度范围为4.5-11mM。
此外,本说明书所用其它术语均具有本领域通用的含义。
为了获得本发明的化合物,需要首先对活性物质进行改造,再将改造后的物质通过连接链连接起来。下面以WEE1抑制剂AZD1775和HSP90抑制剂NVP-AUY922、AT13387和STA9090类似物为例,对一般反应路线进行说明。
首先,如反应式1所示,可按常规的有机化学反应合成方法,将WEE1抑制剂AZD1775改造成含有氨基的化合物2。
接下来,如反应式2所示,可将HSP90抑制剂NVP-AUY922、AT13387和STA9090改造成含有羧基基团的化合物3、5、6,再与前述化合物2缩合。
对于化合物合成的具体方法,将在下文实施例部分进一步详述。
本发明的化合物可用于制备***的药物。具体而言,所述肿瘤包括但不限于胰腺癌、结肠癌、大肠癌、卵巢癌、***、贲门癌、睾丸癌、***癌、肝癌、非小细胞肺癌、小细胞肺癌、肺腺癌、头颈部细胞癌、膀胱癌、胃癌、肾癌、胆管癌、脑癌、乳腺癌、上皮细胞癌、皮肤癌、食管癌、淋巴瘤、神经胶质瘤、黑色素瘤、多发性骨髓瘤和白血病等,包括在其他远离肿瘤原发部位的组织或器官的转移病变。
本发明的化合物可以使用能够产生所需结果的任何方便的手段给药于治疗对象,例如人类患者,例如可以将所述化合物制成如前文所述的药物组合物,和/或制成已知的或新开发的剂型(如片剂、胶囊剂、注射剂等)中。
此外,本发明的化合物可以与其它治疗药物组合使用。所述其它治疗药物包括肿瘤化疗药物、肿瘤靶向药物、肿瘤免疫治疗药物、肿瘤药物偶联物(例如抗体药物偶联物、小分子药物化合物和微型药物化合物)。本发明的化合物可以与其它治疗药物制成同一剂型,或分别制成单独的剂型。
本发明的更具体的实施方式将通过以下实施例并结合附图进行例示性解释说明,但应认识到这些实施例并非意在限制本发明的范围。
实施例
本说明书实施例中所用的合成原料、试验物质等,均为本领域技术人员所公知并且可通过市售或文献方法获得的物质。所用的试验或表征方法也是本领域技术人员公知的方法。
在下文各中间体实施例中,作为起始物质的A1至A4的来源分别参见以下文献:J.Med.Chem.2020,63,5421-5441;Cell Chemical Biology 2019,27,57-65;J.Med.Chem.2014,57,2258-2274;Mol Cancer Ther 2020 19(8):1613–1622.
在下文各药理实施例中,将产物实施例1-4中获得的化合物HTT-19~HTT-30称为“本发明的化合物”。另外,各药理实施例中所用的细胞株来源于:中国科学院大学上海药物研究所(SIMM)。
中间体实施例1
将化合物A1(1.0g,1.7mmol)溶解于20mL体积比为20%的TFA/DCM溶液中,室温条件下,反应1.5h。待反应结束后,减压除去溶剂,得到粗产物。粗产物MTBE打浆,得到目标产物2,黄绿色固体600mg,收率72%。
中间体实施例2
将化合物A2(2.0g,3.2mmol)溶于20mL甲醇中,加入10%的钯碳(200mg),置换氮气和氢气,室温下反应8h。待反应完全后,硅藻土抽滤,减压除去溶剂,得到粗产物。粗产物用硅胶柱层析分离,得到目标产物3,灰白色固体1.35g,收率85%。1H NMR(400MHz,DMSO-d6)δ10.62(s,2H),9.78(s,1H),9.68(s,1H),7.69(s,2H),7.37–7.16(m,5H),6.80(s,2H),6.74(s,1H),3.47(s,4H),3.19(m,1H),2.40(s,4H),1.17(d,J=6.9Hz,6H).13C NMR(100MHz,DMSO-d6)δ173.2,161.9,158.3,151.9,132.46,132.1,130.9,129.1,122.2,114.5,100.8,97.8,66.0,62.3,53.0,26.2,22.4.
中间体实施例3
氮气条件下,将化合物A3(5.0g,9.58mmol)溶解于50mL的甲醇中,加入1.5g 10%Pd/C,置换氢气,室温条件下,反应4h。待反应完全后,垫硅藻土抽滤,除去钯碳。滤液减压除去,得到粗产物。粗产物用DCM和甲醇(10:1)打浆,得到化合物4,黄色固体2.54g,收率85%。1H NMR(400MHz,DMSO-d6)δ9.89(s,1H),9.77(s,1H),7.65(d,J=0.6Hz,1H),7.27(d,J=7.5Hz,1H),6.82–6.66(m,2H),6.52(s,1H),4.66(dd,J=35.0,5.3Hz,2H),4.21(t,J=7.1Hz,2H),3.31–2.97(m,3H),1.22(d,J=6.8Hz,6H).13C NMR(100MHz,DMSO-d6)δ168.12,161.88,157.58,145.00,135.15,131.62,128.07,127.30,119.12,115.05,113.96,112.23,101.86,48.07,27.13,26.65,22.38.
中间体实施例4
将化合物4(2.0g,6.41mmol)和丁二酸酐(774mg,7.69mmol)溶解于20mL的甲苯中,加热至回流,反应12h。待反应完全后,减压除去溶剂,得到粗产物。粗产物用硅胶柱层析,得到化合物5,淡黄色固体2.24g,收率85%。1H NMR(400MHz,DMSO-d6)δ9.89(s,1H),9.72(s,1H),9.19(s,1H),7.65(d,J=0.6Hz,1H),7.56(dd,J=7.5,1.5Hz,1H),7.34(q,J=1.1Hz,1H),7.30(d,J=7.5Hz,1H),6.53(s,1H),4.30–4.05(m,2H),3.16(dtd,J=18.9,7.0,0.9Hz,3H),2.68–2.59(m,2H),2.59–2.48(m,2H),1.22(d,J=6.8Hz,6H).13C NMR(100MHz,DMSO-d6)δ175.04,170.94,167.33,161.76,158.59,138.62,137.57,130.15,127.67,126.80,119.68,117.32,116.56,113.66,101.77,47.57,31.77,28.84,27.00,22.43.中间体实施例5
将化合物A4(1.0g,1.73mmol)溶解于20mL体积比为20%的TFA/DCM溶液中,室温条件下,反应2h。待反应结束后,减压除去溶剂,得到目标产物6,黄色固体750mg,收率83%。
中间体实施例6:7a-7d的通用合成路线
将化合物2(100mg,206μmol)、2a~2d(1.1eq)依次加入单口瓶中,加入无水DMF溶液搅拌并溶解后,滴加DIPEA(3eq)。滴毕,反应体系在室温下搅拌过夜。检测反应完全后,加水淬灭,乙酸乙酯萃取得粗产品,干燥后硅胶柱层析分离纯化,得到相应的产物7a~7d。
化合物7a的合成
以化合物2和2a为原料,按照中间体实施例6的通用合成方法得到化合物7a,淡黄色固体150mg,收率70%。1H NMR(400MHz,CDCl3)δ8.82(s,1H),7.85(d,J=7.8Hz,1H),7.75(d,J=8.0Hz,1H),7.46(d,J=8.7Hz,2H),7.31(d,J=7.6Hz,1H),6.95(d,J=9.0Hz,2H),5.72(ddd,J=16.9,6.1,4.1Hz,1H),5.04(d,J=10.2Hz,2H),4.93(dd,J=17.1,1.0Hz,1H),4.74(d,J=6.1Hz,2H),3.29(d,J=4.7Hz,2H),3.21–3.17(m,4H),2.64(s,4H),2.54(t,J=5.5Hz,2H),1.58(s,6H),1.46(s,9H).
化合物7b的合成
以化合物2和2b为原料,按照中间体实施例6的通用合成方法得到化合物7b,淡黄色固体180mg,收率82%。1H NMR(400MHz,DMSO-d6)δ10.17(s,1H),8.85(d,J=1.1Hz,1H),8.05(s,1H),7.75(d,J=7.4Hz,1H),7.65–7.45(m,3H),6.94(d,J=7.6Hz,2H),6.83(s,1H),5.65(dd,J=14.5,8.9Hz,1H),5.33(s,1H),4.99(d,J=10.2Hz,1H),4.82(d,J=17.3Hz,1H),4.72–4.65(m,2H),3.16–3.02(m,4H),2.94(t,J=12.4Hz,2H),2.51(s,4H),2.37–2.24(m,2H),1.59(t,J=12.0Hz,2H),1.46(s,6H),1.36(s,9H).
化合物7c的合成
以化合物2和2c为原料,按照中间体实施例6的通用合成方法得到化合物7c,淡黄色固体170mg,收率80%。1H NMR(400MHz,DMSO-d6)δ10.15(s,1H),8.84(s,1H),8.04(s,1H),7.75(d,J=7.4Hz,1H),7.67–7.49(m,3H),7.00–6.81(m,3H),5.75–5.59(m,1H),5.33(s,1H),4.97(d,J=10.3Hz,1H),4.81(d,J=17.1Hz,1H),4.69(t,J=6.8Hz,2H),3.17–3.01(m,4H),2.90(t,J=13.4Hz,2H),2.51(s,4H),2.37–2.23(m,2H),1.47(s,6H),1.43–1.33(m,13H).
化合物7d的合成
以化合物2和2d为原料,按照中间体实施例6的通用合成方法得到化合物7d,淡黄色固体150mg,收率75%。1H NMR(400MHz,CDCl3)δ8.88(s,1H),7.86(t,J=7.8Hz,1H),7.75(d,J=8.1Hz,1H),7.46(d,J=8.5Hz,2H),7.33(d,J=7.7Hz,1H),6.92(d,J=8.9Hz,2H),5.73–5.66(m,1H),5.04(d,J=10.0Hz,1H),4.93(d,J=17.2Hz,1H),4.74(d,J=6.1Hz,2H),3.96(s,1H),3.21(s,4H),3.13(d,J=6.6Hz,2H),2.64(s,4H),2.44–2.39(m,2H),1.58(s,6H),1.54–1.50(m,3H),1.44(s,9H),1.37(d,J=7.1Hz,3H).
中间体实施例7:化合物8a~8d的通用合成方法
将化合物7a~7d(1eq)加入到三氟醋酸和二氯甲烷(1∶1比例)的混合溶液中,室温搅拌3小时。待检测反应完全后,减压浓缩,得到黄色固体化合物8a~8d。
中间体实施例8
将化合物2(1eq)、8(1.1eq)依次加入两口瓶中,在氮气保护下注射加入无水DCM溶液,搅拌并溶解后,滴加DIPEA(2eq)。滴毕,反应体系在室温下搅拌3小时。检测反应完全后,加水淬灭,乙酸乙酯萃取得粗产品,减压浓缩后得到产物9,淡黄色固体200mg,收率89%。
中间体实施例9
将化合物9(1eq)、10(1eq)依次加入两口瓶中,在氮气保护下注射加入无水DCM,搅拌并溶解后,滴加DIPEA(3eq)。滴毕,反应体系在室温下搅拌过夜。检测反应完全后,加水淬灭,乙酸乙酯萃取得粗产品,干燥后硅胶柱层析分离纯化,得到相应的产物11,淡黄色固体150mg,收率97%。1H NMR(400MHz,CDCl3)δ8.82(s,1H),7.86(t,J=7.8Hz,1H),7.74(d,J=8.1Hz,1H),7.55(d,J=8.4Hz,2H),7.38(d,J=7.6Hz,1H),7.13(d,J=8.1Hz,2H),7.05(d,J=8.2Hz,2H),6.95(d,J=8.7Hz,2H),5.74–5.64(m,1H),5.03(d,J=10.2Hz,1H),4.92(d,J=17.1Hz,1H),4.74(d,J=6.1Hz,2H),4.60(s,1H),3.79(d,J=38.4Hz,4H),3.36(d,J=6.2Hz,2H),3.22(d,J=4.4Hz,4H),2.78(t,J=6.6Hz,2H),1.58(s,6H),1.43(s,9H).13CNMR(101MHz,CDCl3)δ165.86,162.57,161.20,156.34,155.91,153.78,149.81,147.85,147.46,138.86,136.20,131.56,131.35,129.69,121.79,119.09,117.33,116.20,116.12,79.28,77.39,77.08,76.76,72.45,49.95,47.68,41.76,35.57,30.57,28.43.
中间体实施例10
将化合物11溶于三氟乙酸与DCM(1:1)的溶液中,室温搅拌3小时,监测反应完全后减压浓缩得到12。
产物实施例1:化合物HTT-19;HTT-22~HTT-24的通用合成方法
将化合物8a~8d(1eq)、5(1eq)和HATU(1.2eq)依次加入两口瓶中,在氮气保护下注射加入无水DMF溶液,搅拌并溶解后,滴加DIPEA(3eq)。滴毕,反应体系在室温下搅拌过夜。检测反应完全后,加水淬灭,乙酸乙酯萃取得粗产品,干燥后硅胶柱层析分离纯化,得到相应的产物HTT-19;HTT-22~HTT-24。
化合物HTT-19的合成
以化合物8c和5为原料,按照化合物HTT-19;HTT-22~HTT-24的通用合成方法得到黄色固体HTT-19,淡黄色固体25mg,收率76%。1H NMR(400MHz,DMSO-d6)δ9.90(s,1H),9.74(s,1H),9.61(s,1H),8.82(s,1H),8.04(s,1H),7.93(s,1H),7.74(d,J=7.7Hz,1H),7.62–7.55(m,4H),7.27(s,1H),6.96–6.90(m,3H),6.39(s,1H),5.70–5.61(m,1H),5.33(s,1H),4.99(d,J=9.9Hz,1H),4.82(d,J=17.3Hz,1H),4.66(d,J=5.3Hz,2H),3.98(t,J=8.2Hz,2H),3.06(ddd,J=18.9,14.4,7.1Hz,10H),2.63(s,3H),2.55(d,J=6.9Hz,4H),2.39(t,J=6.9Hz,2H),1.63(s,2H),1.46(s,6H),1.10(d,J=6.9Hz,8H).13C NMR(101MHz,DMSO-d6)δ171.50,170.58,168.04,167.77,161.62,161.44,160.93,157.15,156.49,153.48,147.52,139.30,138.66,135.60,133.42,132.67,125.99,121.67,118.74,117.73,116.74,116.33,116.04,115.64,102.88,72.79,55.39,52.98,49.27,47.06,38.84,32.22,30.92,29.51,28.01,26.31,24.66,23.12.HRMS(ESI):m/z calcd for(M+H)+:952.2103;found:952.2010.
化合物HTT-22的合成
以化合物8a和5为原料,按照化合物HTT-19;HTT-22~HTT-24的通用合成方法得到黄色固体HTT-22,淡黄色固体28mg,收率70%。1H NMR(400MHz,DMSO-d6)δ9.88(s,1H),9.73(s,1H),9.60(s,1H),8.82(s,1H),8.04(s,1H),7.86(s,1H),7.75(d,J=7.6Hz,1H),7.62–7.54(m,4H),7.26(s,1H),6.96(s,1H),6.92(d,J=8.7Hz,2H),6.39(s,1H),5.66(dd,J=16.9,10.4Hz,1H),5.33(s,1H),4.98(d,J=10.0Hz,1H),4.88(d,J=17.4Hz,1H),4.65(d,J=5.0Hz,2H),3.94(t,J=8.4Hz,2H),3.21(d,J=5.6Hz,2H),3.11–3.00(m,7H),2.55(d,J=6.8Hz,5H),2.43(t,J=6.7Hz,4H),1.46(s,6H),1.11(d,J=6.9Hz,6H).13C NMR(101MHz,DMSO-d6)δ171.50,170.58,168.04,167.77,161.62,161.44,160.93,157.15,156.49,153.48,147.52,139.30,138.66,135.60,133.42,132.67,125.99,121.67,118.74,117.73,116.74,116.33,116.04,115.64,102.88,72.79,55.39,52.98,49.27,47.06,38.84,32.22,30.92,29.51,28.01,26.31,24.66,23.10.HRMS(ESI):m/z calcd for(M+H)+:924.1668;found:924.1510.
化合物HTT-23的合成
以化合物8b和5为原料,按照化合物HTT-19;HTT-22~HTT-24的通用合成方法得到黄色固体HTT-23,淡黄色固体30mg,收率75%。1H NMR(400MHz,DMSO-d6)δ9.90(s,1H),9.74(s,1H),9.61(s,1H),8.82(s,1H),8.04(s,1H),7.93(s,1H),7.74(d,J=7.7Hz,1H),7.62–7.55(m,4H),7.27(s,1H),6.96–6.90(m,3H),6.39(s,1H),5.70–5.61(m,1H),5.33(s,1H),4.99(d,J=9.9Hz,1H),4.82(d,J=17.3Hz,1H),4.68(d,J=5.3Hz,2H),3.94(t,J=8.2Hz,2H),3.06(ddd,J=18.9,14.4,7.1Hz,10H),2.63(s,3H),2.55(d,J=6.9Hz,4H),2.39(t,J=6.9Hz,2H),1.63(s,2H),1.46(s,6H),1.10(d,J=6.9Hz,6H).13C NMR(101MHz,DMSO-d6)δ171.50,170.58,168.04,167.77,161.62,161.44,160.93,157.15,156.49,153.48,147.52,139.30,138.66,135.60,133.42,132.67,125.99,121.67,118.74,117.73,116.74,116.33,116.04,115.64,102.88,72.79,55.39,52.98,49.28,47.06,38.84,32.22,30.92,29.51,28.01,26.31,24.66,23.11.HRMS(ESI):m/z calcd for(M+H)+:938.1588;found:938.1308.
化合物HTT-24的合成
以化合物8d和5为原料,按照化合物HTT-19;HTT-22~HTT-24的通用合成方法得到黄色固体HTT-24,淡黄色固体40mg,收率65%。1H NMR(400MHz,DMSO-d6)δ10.17(s,1H),9.90(s,1H),9.76(s,1H),9.63(s,1H),8.83(s,1H),8.04(s,1H),7.86(s,1H),7.75(d,J=7.4Hz,1H),7.63–7.55(m,4H),7.27(s,1H),6.96(s,1H),6.92(d,J=8.2Hz,2H),6.41(s,1H),5.71–5.62(m,1H),5.34(s,1H),4.99(d,J=10.2Hz,1H),4.82(d,J=17.1Hz,1H),4.69(s,2H),3.95(t,J=8.0Hz,2H),3.35(s,6H),3.06(dt,J=28.4,7.3Hz,9H),2.53(s,2H),2.39(t,J=6.7Hz,2H),1.46(s,6H),1.40(d,J=6.9Hz,3H),1.29–1.22(m,3H),1.10(d,J=6.7Hz,6H).13C NMR(101MHz,DMSO-d6)δ171.50,170.58,168.04,167.77,161.62,161.44,160.93,157.15,156.49,153.48,147.52,139.30,138.66,135.60,133.42,132.67,125.99,21.67,118.74,117.73,116.74,116.33,116.04,115.64,102.88,72.79,55.39,52.98,49.27,47.06,38.84,32.22,30.92,29.51,28.01,26.31,24.66,23.12.HRMS(ESI):m/zcalcd for(M+H)+:966.1850;found:966.1535.
产物实施例2:化合物HTT-20;HTT-21;HTT-25;HTT-26的通用合成方法
将化合物8a~8d(1eq)、3(1eq)和HATU(1.2eq)依次加入两口瓶中,在氮气保护下注射加入无水DMF溶液,搅拌并溶解后,滴加DIPEA(3eq)。滴毕,反应体系在室温下搅拌过夜。检测反应完全后,加水淬灭,乙酸乙酯萃取得粗产品,干燥后硅胶柱层析分离纯化,得到相应的产物HTT-20;HTT-21;HTT-25;HTT-26。
化合物HTT-25的合成
以化合物8a和3为原料,按照化合物HTT-20;HTT-21;HTT-25;HTT-26的通用合成方法得到黄色固体HTT-25,淡黄色固体30mg,收率75%。1H NMR(400MHz,DMSO-d6)δ9.74(d,J=41.8Hz,2H),8.88–8.76(m,2H),8.06–7.93(m,1H),7.76(d,J=7.9Hz,1H),7.62(t,J=11.7Hz,3H),7.42(dd,J=36.8,7.7Hz,1H),7.20(q,J=8.1Hz,4H),6.94(d,J=8.7Hz,2H),6.71(s,1H),6.45(s,1H),5.65(ddd,J=16.4,10.5,5.8Hz,1H),5.34(s,1H),4.99(d,J=10.1Hz,1H),4.82(d,J=17.2Hz,1H),4.69(d,J=3.9Hz,2H),3.49(s,4H),3.41(d,J=5.4Hz,2H),3.32(s,2H),3.11(s,4H),2.99–2.93(m,1H),2.59(d,J=29.1Hz,6H),2.27(s,4H),1.46(s,6H),0.88(d,J=6.8Hz,6H).13C NMR(101MHz,DMSO-d6)δ168.03,166.93,161.64,161.41,160.95,160.49,158.07,157.92,156.48,155.23,147.53,143.30,139.30,137.02,132.67,131.40,129.42,129.25,128.94,128.23,127.25,126.06,124.71,119.47,118.73,116.74,115.94,115.21,110.27,104.91,103.19,72.79,66.47,62.43,56.92,53.44,52.96,49.07,48.83,47.07,36.56,30.93,25.82,22.83.HRMS(ESI):m/z calcd for(M+H)+:1070.2588;found:1070.2302.
化合物HTT-21的合成
以化合物8b和3为原料,按照化合物HTT-20;HTT-21;HTT-25;HTT-26的通用合成方法得到黄色固体HTT-21,淡黄色固体35mg,收率70%。1H NMR(400MHz,DMSO-d6)δ10.17(s,1H),9.72(d,J=42.7Hz,2H),8.83(s,2H),8.04(s,1H),7.76(s,1H),7.60(d,J=7.4Hz,3H),7.21(dd,J=18.5,7.3Hz,4H),6.92(d,J=7.4Hz,2H),6.72(s,1H),6.45(s,1H),5.66(d,J=6.1Hz,1H),5.33(s,1H),4.99(d,J=9.9Hz,1H),4.82(d,J=17.0Hz,1H),4.68(s,2H),3.54(s,4H),3.41(s,2H),3.30(s,5H),3.24–2.94(m,8H),2.32(s,4H),1.46(s,4H),1.33–1.20(m,4H),0.89(d,J=6.4Hz,6H).13C NMR(101MHz,DMSO-d6)δ168.04,166.68,161.63,161.43,160.96,160.51,158.39,157.91,156.49,155.20,147.56,139.29,137.36,132.67,131.36,129.28,129.17,128.95,128.21,126.07,121.66,118.75,116.74,115.08,104.96,103.20,72.79,66.60,62.62,53.61,53.17,49.00,47.01,39.14,30.93,29.12,25.83,24.65,22.84.HRMS(ESI):m/z calcd for(M+H)+:964.1630;found:964.1304.
化合物HTT-20的合成
以化合物8c和3为原料,按照化合物HTT-20;HTT-21;HTT-25;HTT-26的通用合成方法得到黄色固体HTT-20,淡黄色固体38mg,收率63%。1H NMR(400MHz,DMSO-d6)δ10.13(s,1H),9.72(d,J=42.7Hz,2H),8.83(s,2H),8.04(s,1H),7.76(s,1H),7.60(d,J=7.4Hz,3H),7.22(dd,J=18.5,7.3Hz,4H),6.92(d,J=7.4Hz,2H),6.72(s,1H),6.45(s,1H),5.66(d,J=6.1Hz,1H),5.33(s,1H),4.99(d,J=9.9Hz,1H),4.82(d,J=17.0Hz,1H),4.68(s,2H),3.54(s,4H),3.41(s,2H),3.28(s,5H),3.24–2.94(m,8H),2.32(s,4H),1.36(s,6H),1.33–1.20(m,4H),0.85(d,J=6.4Hz,6H).13C NMR(101MHz,DMSO-d6)δ168.04,166.68,161.63,161.43,160.96,160.51,158.39,157.91,156.49,155.20,147.56,139.29,137.36,132.67,131.36,129.28,129.17,128.95,128.21,126.07,121.66,118.76,116.75,115.08,104.95,103.20,72.79,66.61,62.62,53.61,53.17,49.00,47.01,39.14,30.93,29.10,25.83,24.60,22.74.HRMS(ESI):m/z calcd for(M+H)+:978.1896;found:978.1633.
化合物HTT-26的合成
以化合物8d和3为原料,按照化合物HTT-20;HTT-21;HTT-25;HTT-26的通用合成方法得到黄色固体HTT-26,淡黄色固体40mg,收率60%。1H NMR(400MHz,DMSO-d6)δ10.17(s,1H),9.72(d,J=42.7Hz,2H),8.83(s,2H),8.04(s,1H),7.76(s,1H),7.60(d,J=7.4Hz,3H),7.21(dd,J=18.5,7.3Hz,4H),6.92(d,J=7.4Hz,2H),6.72(s,1H),6.45(s,1H),5.66(d,J=6.1Hz,1H),5.33(s,1H),4.99(d,J=9.9Hz,1H),4.82(d,J=17.0Hz,1H),4.68(s,2H),3.54(s,4H),3.41(s,2H),3.35(s,5H),3.24–2.94(m,8H),2.32(s,4H),1.46(s,8H),1.33–1.20(m,4H),0.89(d,J=6.4Hz,6H).13C NMR(101MHz,DMSO-d6)δ168.04,166.68,161.63,161.43,160.96,160.51,158.39,157.91,156.49,155.20,147.56,139.29,137.35,132.67,131.36,129.28,129.17,128.95,128.21,126.07,121.66,118.74,116.74,115.99,115.08,104.96,103.20,72.79,66.62,62.62,53.61,53.17,49.00,47.07,39.14,30.93,29.12,25.83,24.65,22.80.HRMS(ESI):m/z calcd for(M+H)+:950.1862;found:950.1211.
产物实施例3:化合物HTT-27的合成
将化合物12(1eq)、3(1eq)和HATU(1.2eq)依次加入两口瓶中,在氮气保护下注射加入无水DMF溶液,搅拌并溶解后,滴加DIPEA(3eq)。滴毕,反应体系在室温下搅拌过夜。检测反应完全后,加水淬灭,乙酸乙酯萃取得粗产品,干燥后硅胶柱层析分离纯化,得到黄色固体产物HTT-27,淡黄色固体25mg,收率70%。1H NMR(400MHz,DMSO-d6)δ9.77(s,1H),9.66(s,1H),8.95(t,J=5.4Hz,1H),8.84(s,1H),8.12–7.94(m,2H),7.74(dd,J=19.7,8.1Hz,2H),7.61(d,J=7.5Hz,2H),7.47(dt,J=15.2,7.4Hz,2H),7.23(d,J=2.4Hz,1H),7.22(s,1H),7.15(d,J=8.0Hz,2H),7.07(d,J=8.3Hz,2H),6.98(d,J=8.8Hz,2H),6.72(s,1H),6.44(s,1H),5.66(ddd,J=16.5,11.0,5.9Hz,1H),5.34(s,1H),4.99(d,J=10.1Hz,1H),4.82(d,J=17.3Hz,1H),4.69(s,2H),4.11(s,1H),3.75(s,2H),3.56(s,6H),3.47(s,5H),3.17(s,6H),3.00–2.93(m,1H),2.80(t,J=6.8Hz,2H),2.37(s,4H),1.46(s,6H),0.89(d,J=6.8Hz,6H).13C NMR(101MHz,DMSO-d6)δ168.06,166.88,161.61,161.44,160.90,160.45,158.08,157.95,156.51,155.22,153.46,150.02,147.53,147.37,143.30,139.29,136.56,136.48,132.65,132.03,129.91,129.40,129.14,128.24,127.20,126.10,124.69,122.13,119.44,118.74,116.84,115.19,110.26,104.92,103.21,72.81,66.34,62.34,55.36,53.39,49.38,47.06,34.41,30.92,25.86,22.84.HRMS(ESI):m/z calcd for(M+H)+:992.1903;found:992.1020.
产物实施例4:化合物HTT-28~HTT-30的通用合成方法
将化合物8a~8d(1eq)、6(1eq)和HATU(1.2eq)依次加入两口瓶中,在氮气保护下注射加入无水DMF溶液,搅拌并溶解后,滴加DIPEA(3eq)。滴毕,反应体系在室温下搅拌过夜。检测反应完全后,加水淬灭,乙酸乙酯萃取得粗产品,干燥后硅胶柱层析分离纯化,得到相应的产物HTT-28~HTT-30。
化合物HTT-28的合成
以化合物8a和6为原料,按照化合物HTT-28~HTT-30的通用合成方法得到黄色固体HTT-28,淡黄色固体30mg,收率75%。1H NMR(400MHz,DMSO-d6)δ10.59(s,1H),10.15(s,1H),9.75(s,1H),8.93(t,J=5.8Hz,1H),8.80(s,1H),8.03(s,1H),7.73(d,J=7.8Hz,1H),7.67(s,1H),7.59(d,J=7.7Hz,3H),7.25(q,J=8.4Hz,4H),6.91(d,J=8.8Hz,2H),6.54(s,1H),6.33(s,1H),5.68–5.62(m,1H),5.32(s,1H),4.98(d,J=9.7Hz,1H),4.81(d,J=17.9Hz,1H),4.67(d,J=4.6Hz,2H),4.05–3.95(m,2H),3.44(s,2H),3.25(d,J=5.5Hz,2H),3.14(d,J=6.6Hz,2H),3.10(s,2H),2.90–2.84(m,3H),2.54(s,4H),2.44(s,6H),1.45(s,6H),1.05(d,J=7.2Hz,3H),0.89(d,J=7.3Hz,3H),0.76(d,J=6.9Hz,6H).13C NMR(101MHz,DMSO-d6)δ169.37,168.04,161.63,161.42,160.93,157.75,156.60,156.53,156.48,154.89,148.17,147.52,139.98,139.28,134.48,132.67,131.41,130.11,129.61,127.76,126.33,125.89,121.66,118.73,116.74,116.02,103.04,102.96,72.79,61.79,57.73,53.33,52.97,48.87,47.07,38.49,37.92,34.96,34.01,30.93,30.29,29.51,27.54,25.72,22.80,21.89,14.89.HRMS(ESI):m/z calcd for(M+H)+:1034.2866;found:1034.1337.
化合物HTT-29的合成
以化合物8c和6为原料,按照化合物HTT-28~HTT-30的通用合成方法得到黄色固体HTT-29,淡黄色固体31mg,收率78%。1H NMR(400MHz,DMSO-d6)δ10.61(s,1H),10.17(s,1H),9.80(s,1H),8.96(t,J=5.5Hz,1H),8.82(s,1H),8.04(s,1H),7.78–7.69(m,2H),7.60(d,J=7.6Hz,3H),7.33(dd,J=31.4,7.9Hz,4H),6.92(d,J=8.2Hz,2H),6.46(d,J=83.1Hz,2H),5.66(dq,J=11.2,6.0Hz,1H),5.34(s,1H),4.99(d,J=10.2Hz,1H),4.82(d,J=17.1Hz,1H),4.69(s,2H),3.50(s,2H),3.36(s,6H),3.15(dd,J=13.7,6.1Hz,8H),2.91(t,J=10.5Hz,3H),2.57(s,2H),2.46(s,6H),1.46(s,8H),1.26–1.21(m,2H),1.03(t,J=7.1Hz,3H),0.79(d,J=6.7Hz,6H).13C NMR(101MHz,DMSO-d6)δ169.37,168.04,161.63,161.42,160.93,157.75,156.60,156.53,156.48,154.89,148.17,147.52,139.98,139.28,134.48,132.67,131.41,130.11,129.61,127.76,126.33,125.89,121.66,118.73,116.74,116.02,103.04,102.96,72.79,61.79,57.73,53.33,52.97,48.87,47.07,38.49,37.92,34.96,34.01,30.93,30.29,29.51,27.64,25.72,22.81,21.89,14.96.HRMS(ESI):m/zcalcd for(M+H)+:1062.2899;found:1062.1903.
化合物HTT-30的合成
以化合物8d和6为原料,按照化合物HTT-28~HTT-30的通用合成方法得到黄色固体HTT-30,淡黄色固体38mg,收率70%。1H NMR(400MHz,DMSO)δ10.62(s,1H),10.18(s,1H),9.86(d,J=10.0Hz,1H),8.97(t,J=5.4Hz,1H),8.82(s,1H),8.04(s,1H),7.77–7.70(m,2H),7.61(d,J=7.5Hz,3H),7.37(d,J=7.8Hz,2H),7.29(d,J=7.8Hz,2H),6.93(d,J=8.0Hz,2H),6.56(s,1H),6.39(d,J=6.2Hz,1H),5.69–5.62(m,1H),5.35(s,1H),4.98(d,J=10.2Hz,1H),4.82(d,J=17.1Hz,1H),4.68(s,2H),3.51(s,2H),3.35(s,8H),3.18–3.14(m,4H),3.09(d,J=6.0Hz,2H),3.04(d,J=7.1Hz,1H),2.92(d,J=8.8Hz,4H),2.67(s,2H),2.46(s,4H),1.46(s,6H),1.23(d,J=11.3Hz,6H),1.03(t,J=7.0Hz,3H),0.79(d,J=6.7Hz,6H).13C NMR(101MHz,DMSO)δ170.06,169.31,168.05,161.61,160.94,157.79,156.62,156.52,154.90,148.17,147.52,139.29,134.49,132.67,130.11,129.62,127.77,126.32,125.87,121.64,118.73,116.76,116.10,103.07,102.94,72.79,61.74,53.28,52.93,47.06,45.83,38.50,37.92,34.96,34.01,31.62,30.92,30.29,29.51,25.71,24.59,22.81,21.89,14.96,8.98.HRMS(ESI):m/z calcd for(M+H)+:1078.3933;found:1078.2003.
药理实施例1:化合物HTT-19、HTT-20和HTT-21的Western实验,以及化合物HTT-21对MV-4-11肿瘤细胞株的增殖抑制活性实验
首先,为评价本发明中率先合成的化合物HTT-19、HTT-20和HTT-21对WEE1蛋白的影响,采用Western实验对化合物进行测试。细胞接种于六孔板中,培养过夜后加不同浓度的化合物处理24小时后收集细胞。预冷PBS洗一次,加入1×SDS上样缓冲液裂解细胞。收集细胞裂解物,沸水浴加热10min后于4℃12000rpm离心5min。取上清液进行SDS-PAGE电泳。电泳结束后并转移,转移结束后,用丽春红染色确定转移情况和蛋白条带在硝酸纤维素膜上的位置,标记后将目的条带用封闭液于摇床室温封闭1h。然后,将膜置于一抗中4摄氏度孵育过夜。用TBST洗涤液室温洗涤三次,每次10min。加入辣根过氧化物酶标记的二抗,摇床室温孵育1h。再用TBST洗涤三次,每次10min后,显色,曝光。
如附图1所示,首先用化合物HTT-19~HTT-21在不同浓度下处理MV-4-11细胞12小时,并进行Western实验,观察WEE1、AKT以及Actin的下***况。对照组DMSO不能引起WEE1、AKT以及Actin的降解,而这三个化合物中HTT-20和HTT-21在0.1微摩尔浓度下即可明显诱导WEE1的下调,且HTT-21在1微摩尔时显示出了与PROTACs相似的虎克效应,对于Actin没有影响,在较高浓度10微摩尔时才可明显诱导AKT下调。这种对WEE1和AKT的下调可能是由HSP90配体引起的客户蛋白降解,且HSP90对这两种客户蛋白降解的差别可能是由于化合物的一端起到了与PROTACs靶蛋白配体相似的作用,即拉近伴侣蛋白与WEE1的距离导致伴侣蛋白介导的UPS对WEE1起到了的标记的作用,初步显示出了本发明中的分子对WEE1的靶点选择性降解。
药理实施例2:化合物HTT-21对肿瘤细胞株的增殖抑制活性实验
接下来,我们对降解能力较强的HTT-21进行抗增殖实验。化合物对肿瘤细胞生长抑制检测采用CCK8方法。具体步骤如下:处于对数生长期的细胞按合适密度接种至96孔培养板,每孔100μL完全培养基培养过夜。加入一系列浓度的化合物,每个浓度设置三个复孔,并设置无化合物作用的阳性对照孔及无细胞阴性对照孔。将细胞培养在37℃条件下72h。化合物作用结束后,每孔加入10μLCCK-8试剂,置于37℃培养箱中放置3小时后,使用全波长式微孔板酶标仪SpectraMax 190测定450nm波长下的光密度(OD值)。附图2显示MV-4-11细胞经过化合物处理3天后,能够表现出明显的抗肿瘤增值活性IC50=8nM。
药理实施例3:化合物HTT-22~HTT-30对肿瘤细胞株的增殖抑制活性实验
基于以上实验结果,为了进一步验证本发明中HSP90抑制剂与WEE1抑制剂形成化合物的生物活性。对化合物HTT-22~HTT-30进行了的肿瘤细胞抑制实验,并以AZD1775作为对照,采用与药理实施例2相同的试验方法。
表1.化合物在体外对MV-4-11肿瘤细胞系的抗增殖活性
a Data is the mean±SD value of three independent determinations.
b Positive control.
如表1所示,在MV-4-11肿瘤细胞上,本发明中大部分化合物能够表现出明显的抗肿瘤活性,并且与AZD1775相比表现出更强的抗肿瘤能力。其中,肿瘤抑制活性最强的化合物HTT-22;HTT-28;HTT-29;HTT-30表现出IC50在8nM到13nM,说明化合物在体外能有效抑制肿瘤。同系列化合物HTT-27(IC50=24.16nM)与HTT-21(IC50=8nM)相比并没有表现出更好的抗肿瘤活性,说明HTT-27中的连接方式可能影响分子的抗肿瘤活性,因此不再合成相关linker化合物。总体来看,烷基链linker越短,活性越高;HSP90配体为STA9090改造后的化合物分子抗肿瘤的活性较好。
药理实施例4:化合物的蛋白免疫印迹实验
为进一步验证本发明分子靶蛋白降解活性和肿瘤细胞抑制活性,并验证同一配体不同链长及不同连接方式对生物活性的影响,对与化合物HTT-19同一系列的化合物HTT-22~HTT-24,和与HTT-20;HTT-21同一系列的HTT-25~HTT-27进行Western实验。使用AT13387、AUY922和STA9090作为阳性对照,采用与药理实施例1相同的试验方法,实验结果见附图3(a;b);图4(a;b);图5(a;b)。
如附图3(a;b)所示,以HSP90抑制剂AT13387作对照,MV-4-11细胞株以不同浓度的化合物HTT-22~HTT-24处理12小时,通过Western实验观察WEE1、AKT以及GAPDH的降解情况。化合物HTT-22~HTT-24均能在0.1~1微摩尔时有选择性的降解WEE1,对AKT降解能力较弱,体现了HSP90-WEE1化合物分子一定的靶蛋白选择性。而AT13387则对WEE1以及AKT均有较强的降解能力,可能的原因是WEE1与AKT一样均是HSP90的客户蛋白,而缺乏选择性。
如附图4(a;b)所示,以HSP90抑制剂AUY922作对照,MV-4-11细胞株用化合物HTT-25~HTT-27在不同浓度下处理12小时后,出现了与HTT-22~24相似的结果,其中化合物HTT-26甚至在0.01微摩尔时表现出了WEE1的选择性降解能力,AUY922也存在一定的WEE1选择性降解能力。两组实验结果表明本发明化合物分子存在一定的WEE1靶蛋白选择性降解能力。基于以上药理实施结果,以HSP90抑制剂STA9090作为对照并对化合物HTT-28~HTT-30进行了Western实验。
如附图5(a;b)所示,HTT-28;HTT-29处理MV-4-11 12小时后同样对WEE1有一定的选择性降解能力。其中化合物HTT-29在0.1微摩尔时即可有效降解WEE1蛋白,在化合物浓度为1微摩尔时同样表现出了虎克效应,而对AKT则没有表现出明显的影响,HSP90抑制剂则几乎没有表现出对WEE1和AKT的选择性。
以不同浓度的WEE1抑制剂处理MV-4-11细胞株12小时,通过Western实验观察抑制剂AZD1775对WEE1蛋白的作用。如附图6结果显示,抑制剂几乎不降解WEE1,说明本发明化合物与抑制剂有着不同的作用机理。
药理实施例5:化合物诱导转录实验
为进一步验证本发明化合物的活性,如附图7所示,以DMSO作为对照,MV-4-11用0.01~1μM四个浓度的HTT-19~HTT-21处理6小时,并观察WEE1转录组的表达水平。实验结果显示,化合物并不能诱导降解WEE1的上游转录组,而是直接诱导降解靶蛋白以下调靶蛋白含量。而附图8WEE1抑制剂以DMSO作为对照,可影响靶蛋白的上游转录组表达,体现了本发明化合物与WEE1抑制剂不同的作用机制。
本发明设计合成了12个HSP90抑制剂与WEE1抑制剂结合的化合物,并进行了一系列体外生物活性研究。
在体外肿瘤细胞抗增殖实验中,化合物均对MV-4-11的抗肿瘤活性较好,普遍优于WEE1抑制剂AZD1775(IC50=67.49nM),其中化合物HTT-21;HTT-22;HTT-28;HTT-29;HTT-30处理MV-4-11 12小时后IC50在8nM到13nM。进一步的Western实验中,该发明的化合物均能诱导降解WEE1蛋白而WEE1抑制剂则对蛋白几乎没有降解作用,其中化合物HTT-21;HTT-22;HTT-28;HTT-29;HTT-30能实现WEE1蛋白的选择性降解而几乎不影响AKT。化合物HTT-21和HTT-29能在0.1μM时降解80%以上的WEE1蛋白,体现了极强的选择性降解能力,说明linker为三或四个烷基碳链时能最大化的提高选择性WEE1降解能力和肿瘤抑制活性。与抑制剂AZD1775相比,HSP90-WEE1化合物分子的作用机制是直接诱导WEE1降解,并不影响上游转录。
本发明得到了一系列HSP90-WEE1化合物分子,在提高了WEE1抑制剂的靶向性的同时,证明了HSP90通过伴侣蛋白的作用在该发明分子中可以介导WEE1的选择性降解,对WEE1起到了双重作用,提高了药物的抗肿瘤活性。
Claims (10)
1.一种化合物,其包括WEE1抑制部分与HSP90抑制部分,两者通过连接链连接;或其药学上可接受的盐、溶剂合物、互变异构体、立体异构体、外消旋体或前药;
2.根据权利要求1所述的化合物,其中所述连接链包含在HSP90抑制部分和WEE1抑制部分之间形成共价结合的键或者连接基团,所述连接链选自:化学键、酯键、羰基、C1-6亚烷基、酰胺键、醚键、二硫键及其组合;
或其药学上可接受的盐、溶剂合物、互变异构体、立体异构体、外消旋体或前药。
3.根据权利要求1或2所述的化合物,其中所述HSP90抑制部分选自如下结构,以及由这些结构衍生而成的能够和连接链形成各类共价键的结构或基团:
或其药学上可接受的盐、溶剂合物、互变异构体、立体异构体、外消旋体或前药。
4.根据权利要求1所述的化合物,其中所述WEE1抑制部分选自如下结构,以及由这些结构衍生而成的能够和连接链形成各类共价键的结构或基团:
或其药学上可接受的盐、溶剂合物、互变异构体、立体异构体、外消旋体或前药。
5.根据权利要求3或4所述的化合物,其中所述HSP90抑制部分包括所述NVP-AUY922或AT13387类似物的结构,或由NVP-AUY922或AT13387类似物衍生的结构;WEE1抑制部分包括所述AZD1775的结构,或由AZD1775衍生的结构;连接链选自化学键、酰胺键、羰基、C1-6亚烷基及其组合;
或其药学上可接受的盐、溶剂合物、互变异构体、立体异构体、外消旋体或前药。
6.根据权利要求3、4或5所述的化合物,其中,
HSP90抑制部分包含从以下结构中除去一个氢原子后得到的一价基团:
其中R1表示-COOH,R2表示-NHC(=O)CH2CH2COOH,R3表示-CH2COOH;
WEE1抑制部分包含从以下结构中除去一个氢原子后得到的一价基团:
连接链选自化学键:-NH-(CH2)n-、-NH-(CH2)2-Ph-O-C(=O)-,
或其药学上可接受的盐、溶剂合物、互变异构体、立体异构体、外消旋体或前药。
7.根据前述任一项权利要求所述的化合物,其为如下所示的HTT-19~HTT-30,
或其药学上可接受的盐、溶剂合物、互变异构体、立体异构体、外消旋体或前药。
8.一种药物组合物,其包含根据前述任一项权利要求所述的化合物或其药学上可接受的盐、溶剂合物、互变异构体、立体异构体、外消旋体或前药,以及一种或多种药用载体。
9.根据权利要求8所述的药物组合物,其还包含其它治疗药物,所述其它治疗药物为肿瘤化疗药物、肿瘤靶向药物、肿瘤免疫治疗药物及肿瘤药物偶联物。
10.一种权利要求1-7中任一项所述的化合物或权利要求8-9中任一项所述的药物组合物用于制备预防或***药物的用途。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202310470542.4A CN116925084A (zh) | 2023-04-27 | 2023-04-27 | 对肿瘤过表达的细胞周期调节蛋白wee1有双重抑制作用的化合物 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202310470542.4A CN116925084A (zh) | 2023-04-27 | 2023-04-27 | 对肿瘤过表达的细胞周期调节蛋白wee1有双重抑制作用的化合物 |
Publications (1)
Publication Number | Publication Date |
---|---|
CN116925084A true CN116925084A (zh) | 2023-10-24 |
Family
ID=88386842
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202310470542.4A Pending CN116925084A (zh) | 2023-04-27 | 2023-04-27 | 对肿瘤过表达的细胞周期调节蛋白wee1有双重抑制作用的化合物 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN116925084A (zh) |
-
2023
- 2023-04-27 CN CN202310470542.4A patent/CN116925084A/zh active Pending
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Rakesh et al. | Anticancer and DNA binding studies of potential amino acids based quinazolinone analogs: Synthesis, SAR and molecular docking | |
CN113603676B (zh) | 基于厄洛替尼靶向降解egfr蛋白小分子化合物及其制备方法和应用 | |
CN112094266B (zh) | 一种吡咯并吡啶酮类化合物、其制备方法、其组合物和用途 | |
CN110563703B (zh) | 基于crbn配体诱导parp-1降解的化合物及制备方法和应用 | |
JP7041821B2 (ja) | アミノ置換窒素含有縮合環化合物、その調製方法及び使用 | |
Li et al. | Discovery of novel β-carboline/acylhydrazone hybrids as potent antitumor agents and overcome drug resistance | |
WO2013170758A1 (zh) | 含稠环结构的苯甲酰胺类化合物及其作为抗肿瘤药物应用 | |
EP3597186B1 (en) | Myc g-quadruplex stabilizing small molecules and their use | |
CN103420923B (zh) | 4-氨基喹唑啉异羟肟酸类化合物及作为抗肿瘤药物应用 | |
WO2022199547A1 (zh) | 一种7,9-二氢嘌呤衍生物及其制药用途 | |
KR102452412B1 (ko) | 트리프톨리드의 c14-히드록실 에스테르화 아미노산 유도체, 및 그의 제조 방법 및 용도 | |
WO2009124468A1 (zh) | 具有高活性的四环蒽醌类抗生素的衍生物及其制备和应用 | |
WO2019034178A1 (zh) | 一种dna毒性二聚体化合物 | |
CN114195814A (zh) | 羟基萘酮-苯硼酸类化合物、制备方法和用途 | |
CN117024413B (zh) | 3-氨基吡嗪-2-甲酰胺类靶向蛋白水解嵌合体及其制备方法、药物组合物和应用 | |
CN114349738B (zh) | 一类靶向降解cdk2的小分子缀合物及其应用 | |
Wu et al. | Development and structure-activity relationship of tacrine derivatives as highly potent CDK2/9 inhibitors for the treatment of cancer | |
Qiu et al. | Design, synthesis and biological evaluation of matrine contains benzimidazole derivatives as dual TOPOI and PARP inhibitors for cancer therapy | |
CN108358894B (zh) | 一种抑制组蛋白乙酰转移酶的化合物及其制备方法与应用 | |
KR19980032504A (ko) | 포유류 세포의 성장 억제 방법 | |
CN116925084A (zh) | 对肿瘤过表达的细胞周期调节蛋白wee1有双重抑制作用的化合物 | |
WO2019157959A1 (zh) | 一种嘧啶类化合物、其制备方法及其医药用途 | |
WO2019230669A1 (ja) | Runx結合配列を標的とする医薬組成物およびrunx阻害剤 | |
CN111247143B (zh) | 可用作蛋白激酶抑制剂的吡啶并喹唑啉衍生物 | |
CN116836179A (zh) | 具有肿瘤靶向的溴结构域蛋白brd4抑制剂 |
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 |