Compound (SMILES): CNC(=O)N1N=C(c2ccc(N3CCN(C(=O)CCCCNC(=O)COc4cccc5c4C(=O)N(C4CCC(=O)NC4=O)C5=O)CC3)cc2)c2cc(OC)c(OC)cc2C[C@@H]1C
Modality: PROteolysis-TArgeting Chimera (PROTAC)
Target protein (POI): BRD4 (UniProt O60885)
POI sequence: MSAESGPGTRLRNLPVMGDGLETSQMSTTQAQAQPQPANAASTNPPPPETSNPNKPKRQTNQLQYLLRVVLKTLWKHQFAWPFQQPVDAVKLNLPDYYKIIKTPMDMGTIKKRLENNYYWNAQECIQDFNTMFTNCYIYNKPGDDIVLMAEALEKLFLQKINELPTEETEIMIVQAKGRGRGRKETGTAKPGVSTVPNTTQASTPPQTQTPQPNPPPVQATPHPFPAVTPDLIVQTPVMTVVPPQPLQTPPPVPPQPQPPPAPAPQPVQSHPPIIAATPQPVKTKKGVKRKADTTTPTTIDPIHEPPSLPPEPKTTKLGQRRESSRPVKPPKKDVPDSQQHPAPEKSSKVSEQLKCCSGILKEMFAKKHAAYAWPFYKPVDVEALGLHDYCDIIKHPMDMSTIKSKLEAREYRDAQEFGADVRLMFSNCYKYNPPDHEVVAMARKLQDVFEMRFAKMPDEPEEPVVAVSSPAVPPPTKVVAPPSSSDSSSDSSSDSDSSTDDSEEERAQRLAELQEQLKAVHEQLAALSQPQQNKPKKKEKDKKEKKKEKHKRKEEVEENKKSKAKEPPPKKTKKNNSSNSNVSKKEPAPMKSKPPPTYESEEEDKCKPMSYEEKRQLSLDINKLPGEKLGRVVHIIQSREPSLKNSNPDEIEIDFETLKPSTLRELERYVTSCLRKKRKPQAEKVDVIAGSSKMKGFSSSESESSSESSSSDSEDSETEMAPKSKKKGHPGREQKKHHHHHHQQMQQAPAPVPQQPPPPPQQPPPPPPPQQQQQPPPPPPPPSMPQQAAPAMKSSPPPFIATQVPVLEPQLPGSVFDPIGHFTQPILHLPQPELPPHLPQPPEHSTPPHLNQHAVVSPPALHNALPQQPSRPSNRAAALPPKPARPPAVSPALTQTPLLPQPPMAQPPQVLLEDEEPPAPPLTSMQMQLYLQQLQKVQPPTPLLPSVKVQSQPPPPLPPPPHPSVQQQLQQQPPPPPPPQPQPPPQQQHQPPPRPVHLQPMQFSTHIQQPPPPQGQQPPHPPPGQQPPPPQPAKPQQVIQHHHSPRHHKSDPYSTGHLREAPSPLMIHSPQMSQFQSLTHQSPPQQNVQPKKQELRAASVVQPQPLVVVKEEKIHSPIIRSEPFSPSLRPEPPKHPESIKAPVHLPQRPEMKPVDVGRPVIRPPEQNAPPPGAPDKDKQKQEPKTPVAPKKDLKIKNMGSWASLVQKHPTTPSSTAKSSSDSFEQFRRAAREKEEREKALKAQAEHAEKEKERLRQERMRSREDEDALEQARRAHEEARRRQEQQQQQRQEQQQQQQQQAAAVAAAATPQAQSSQPQSMLDQQRELARKREQERRRREAMAATIDMNFQSDLLSIFEENLF
E3 ligase: CRBN (UniProt Q96SW2)
Ligase sequence: MAGEGDQQDAAHNMGNHLPLLPAESEEEDEMEVEDQDSKEAKKPNIINFDTSLPTSHTYLGADMEEFHGRTLHDDDSCQVIPVLPQVMMILIPGQTLPLQLFHPQEVSMVRNLIQKDRTFAVLAYSNVQEREAQFGTTAEIYAYREEQDFGIEIVKVKAIGRQRFKVLELRTQSDGIQQAKVQILPECVLPSTMSAVQLESLNKCQIFPSKPVSREDQCSYKWWQKYQKRKFHCANLTSWPRWLYSLYDAETLMDRIKKQLREWDENLKDDSLPSNPIDFSYRVAACLPIDDVLRIQLLKIGSAIQRLRCELDIMNKCTSLCCKQCQETEITTKNEIFSLSLCGPMAAYVNPHGYVHETLTVYKACNLNLIGRPSTEHSWFPGYAWTVAQCKICASHIGWKFTATKKDMSPQKFWGLTRSALLPTIPDTEDEISPDKVILCL
Cell line: HEK293 (Homo sapiens)
Cell line description: Used for the production of the Gamaleya SARS-CoV-2 (COVID-19) vaccine (Trade name
A*02 A*03
In situ; Fetal kidney; 
Vaccine production cell line.
NCBI_TaxID; 28285; Human adenovirus C serotype 5.
Transformed cell line
Assay: High-Content Analysis
Assay time (hours): 15
DC50:  16.005 nM
Assay description: Quantification of degradation of fluorescently labelled BRD4 in HEK293 cells
HEK293 cells stably expressing N-terminal fluorescently tagged BRD4 variants were generated by stable integration of expression constructs into HEK293 wild-type cells. PROTAC-mediated degradation was measured by quantification of the tagGFP2-BRD4 fluorescence intensity after 24h compound treatment. For this, human embryonic kidney HEK293 cells (DSMZ ACC 305) stably expressing tagGFP2-BRD4 were seeded in 20 pi into 384- well microtiter plates (8000 cells per well) containing 200 nl_ of compounds in Dose- Response. Control wells received DMSO. Plates were then incubated for 24h at 37 °C in a humidified incubator and fixed with 1% PFA for 30 min. Cells were then stained with Hoechst 33342 (H-1399, 0.1pg/mL) and imaged on an automated confocal microscopy system. Nuclear and cytoplasmic intensity of the tagGFP2 fluorescence signal was determined by automated image analysis using custom generated scripts (MetaXpress). Data were then transferred to Genedata Screener software, normalized to DMSO and DCso values (half-maximal concentration to achieve maximal reduction of tagGFP staining intensity of the tagGFP2- BRD4 reporter fusion proteins relative to DMSO) are reported.
Reference: SUBSTITUTED 2,3-BENZODIAZEPINES DERIVATIVES (patent)
Source database: TPDdb