Compound (SMILES): O=C(CN1CCN(CCCCCCCc2nc3ccc(N4CCN(C(=O)c5cc(Cc6n[nH]c(=O)c7ccccc67)ccc5F)CC4)nc3[nH]2)CC1)NCCNc1cccc2c1C(=O)N(C1CCC(=O)NC1=O)C2=O
Modality: PROteolysis-TArgeting Chimera (PROTAC)
Target protein (POI): PARP1 (UniProt P09874)
POI sequence: MAESSDKLYRVEYAKSGRASCKKCSESIPKDSLRMAIMVQSPMFDGKVPHWYHFSCFWKVGHSIRHPDVEVDGFSELRWDDQQKVKKTAEAGGVTGKGQDGIGSKAEKTLGDFAAEYAKSNRSTCKGCMEKIEKGQVRLSKKMVDPEKPQLGMIDRWYHPGCFVKNREELGFRPEYSASQLKGFSLLATEDKEALKKQLPGVKSEGKRKGDEVDGVDEVAKKKSKKEKDKDSKLEKALKAQNDLIWNIKDELKKVCSTNDLKELLIFNKQQVPSGESAILDRVADGMVFGALLPCEECSGQLVFKSDAYYCTGDVTAWTKCMVKTQTPNRKEWVTPKEFREISYLKKLKVKKQDRIFPPETSASVAATPPPSTASAPAAVNSSASADKPLSNMKILTLGKLSRNKDEVKAMIEKLGGKLTGTANKASLCISTKKEVEKMNKKMEEVKEANIRVVSEDFLQDVSASTKSLQELFLAHILSPWGAEVKAEPVEVVAPRGKSGAALSKKSKGQVKEEGINKSEKRMKLTLKGGAAVDPDSGLEHSAHVLEKGGKVFSATLGLVDIVKGTNSYYKLQLLEDDKENRYWIFRSWGRVGTVIGSNKLEQMPSKEDAIEHFMKLYEEKTGNAWHSKNFTKYPKKFYPLEIDYGQDEEAVKKLTVNPGTKSKLPKPVQDLIKMIFDVESMKKAMVEYEIDLQKMPLGKLSKRQIQAAYSILSEVQQAVSQGSSDSQILDLSNRFYTLIPHDFGMKKPPLLNNADSVQAKVEMLDNLLDIEVAYSLLRGGSDDSSKDPIDVNYEKLKTDIKVVDRDSEEAEIIRKYVKNTHATTHNAYDLEVIDIFKIEREGECQRYKPFKQLHNRRLLWHGSRTTNFAGILSQGLRIAPPEAPVTGYMFGKGIYFADMVSKSANYCHTSQGDPIGLILLGEVALGNMYELKHASHISKLPKGKHSVKGLGKTTPDPSANISLDGVDVPLGTGISSGVNDTSLLYNEYIVYDIAQVNLKYLLKLKFNFKTSLW
E3 ligase: CRBN (UniProt Q96SW2)
Ligase sequence: MAGEGDQQDAAHNMGNHLPLLPAESEEEDEMEVEDQDSKEAKKPNIINFDTSLPTSHTYLGADMEEFHGRTLHDDDSCQVIPVLPQVMMILIPGQTLPLQLFHPQEVSMVRNLIQKDRTFAVLAYSNVQEREAQFGTTAEIYAYREEQDFGIEIVKVKAIGRQRFKVLELRTQSDGIQQAKVQILPECVLPSTMSAVQLESLNKCQIFPSKPVSREDQCSYKWWQKYQKRKFHCANLTSWPRWLYSLYDAETLMDRIKKQLREWDENLKDDSLPSNPIDFSYRVAACLPIDDVLRIQLLKIGSAIQRLRCELDIMNKCTSLCCKQCQETEITTKNEIFSLSLCGPMAAYVNPHGYVHETLTVYKACNLNLIGRPSTEHSWFPGYAWTVAQCKICASHIGWKFTATKKDMSPQKFWGLTRSALLPTIPDTEDEISPDKVILCL
Cell line: SW620 (Homo sapiens)
Cell line description: African=2.02%; Native American=0%; East Asian, North=4.3%; East Asian, South=0%; South Asian=0%; European, North=50.01%; European, South=43.67%.
Mutation; HGNC; HGNC Mutation; HGNC; HGNC Mutation; HGNC; HGNC Mutation; HGNC; HGNC
A*02 A*02,24; B*07,15; C*07 A*02
Stable (MSS).
Metastatic; Lymph node; 
Caucasian.
Cancer cell line
Assay: Nano-Glo HiBiT
DC50:  47.7 nM
Assay description: PARP1 protein degradation assay
Sw620 colon cancer cells were cultured in Leibovitz's L-15 medium (Gibco). HiBiT-PARP1 plasmid was transfected into the cells using Lipofectamine 2000 (Thermo Fisher) and incubated for 24 hours at 37°C in a CO2-free incubator. Cells were harvested and frozen in aliquots at -80°C in a freezing solution of 10% DMSO and 90% FBS. An aliquot of the frozen SW620/HiBiT-PARP1 cells was quickly placed in a 37°C water bath and allowed to lyse completely. After centrifugation, the cells were resuspended in Leibovitz's L-15 medium at a cell density of 5.0 x 104 cells/mL. Using a Multidrop combi (Thermo Scientific), 20 μL of cells were added to a 384-well compound plate containing 20 μL of compound. The cell density was 1000 cells/well. The cells were incubated for 24 hours at 37°C in a CO2-free incubator. After equilibration at room temperature for 10 minutes, 20 nL of Nano-Glo HiBiT (Promega) lysis reagent was added using a Multidrop Combi. The mixture was incubated at room temperature for 10 minutes, and the signal was read using EnVision (Perkin Elmer). The fluorescence signal (RFU) was collected using Envision. The maximum RFU value was used to calculate the compound degradation effect for each well on the assay plate according to the following formula: %Effect = 100 × (Value - ZPE) / (HPE - ZPE). Where %Effect is the compound degradation effect for the corresponding well, Value is the signal value for that well, ZPE is the mean signal value for the negative control wells, and HPE is the mean signal value for the positive control wells. The positive control was 1 μM SK575. The test compound concentration was diluted 3.162-fold to 11 concentrations ranging from 10 μM to 0.1 nM. XLFit was used to fit the compound percentage degradation rate and the 11-point concentration data to a parametric nonlinear logistic equation to calculate the compound DC50 value.
Reference: PROTEIN DEGRADATION AGENT AND PREPARATION METHOD THEREFOR AND PHARMACEUTICAL USE THEREOF (patent)
Source database: TPDdb