ailab-bio/TACK huggingface dataset:
Compound (SMILES): Nc1nccn2c(N3CCC4(CC3)CN(CCOCCCCCOc3ccc5c(c3)CN(C3CCC(=O)NC3=O)C5=O)C4)nc(-c3ccc(C(=O)Nc4cc(Cl)ccn4)cc3F)c12
Modality: PROteolysis-TArgeting Chimera (PROTAC)
Target protein (POI): BTK (UniProt Q06187)
POI sequence: MAAVILESIFLKRSQQKKKTSPLNFKKRLFLLTVHKLSYYEYDFERGRRGSKKGSIDVEKITCVETVVPEKNPPPERQIPRRGEESSEMEQISIIERFPYPFQVVYDEGPLYVFSPTEELRKRWIHQLKNVIRYNSDLVQKYHPCFWIDGQYLCCSQTAKNAMGCQILENRNGSLKPGSSHRKTKKPLPPTPEEDQILKKPLPPEPAAAPVSTSELKKVVALYDYMPMNANDLQLRKGDEYFILEESNLPWWRARDKNGQEGYIPSNYVTEAEDSIEMYEWYSKHMTRSQAEQLLKQEGKEGGFIVRDSSKAGKYTVSVFAKSTGDPQGVIRHYVVCSTPQSQYYLAEKHLFSTIPELINYHQHNSAGLISRLKYPVSQQNKNAPSTAGLGYGSWEIDPKDLTFLKELGTGQFGVVKYGKWRGQYDVAIKMIKEGSMSEDEFIEEAKVMMNLSHEKLVQLYGVCTKQRPIFIITEYMANGCLLNYLREMRHRFQTQQLLEMCKDVCEAMEYLESKQFLHRDLAARNCLVNDQGVVKVSDFGLSRYVLDDEYTSSVGSKFPVRWSPPEVLMYSKFSSKSDIWAFGVLMWEIYSLGKMPYERFTNSETAEHIAQGLRLYRPHLASEKVYTIMYSCWHEKADERPTFKILLSNILDVMDEES
E3 ligase: CRBN (UniProt Q96SW2)
Ligase sequence: MAGEGDQQDAAHNMGNHLPLLPAESEEEDEMEVEDQDSKEAKKPNIINFDTSLPTSHTYLGADMEEFHGRTLHDDDSCQVIPVLPQVMMILIPGQTLPLQLFHPQEVSMVRNLIQKDRTFAVLAYSNVQEREAQFGTTAEIYAYREEQDFGIEIVKVKAIGRQRFKVLELRTQSDGIQQAKVQILPECVLPSTMSAVQLESLNKCQIFPSKPVSREDQCSYKWWQKYQKRKFHCANLTSWPRWLYSLYDAETLMDRIKKQLREWDENLKDDSLPSNPIDFSYRVAACLPIDDVLRIQLLKIGSAIQRLRCELDIMNKCTSLCCKQCQETEITTKNEIFSLSLCGPMAAYVNPHGYVHETLTVYKACNLNLIGRPSTEHSWFPGYAWTVAQCKICASHIGWKFTATKKDMSPQKFWGLTRSALLPTIPDTEDEISPDKVILCL
Cell line: OCI-Ly10 (Homo sapiens)
Cell line description: African=0.25%; Native American=0.93%; East Asian, North=0.48%; East Asian, South=0%; South Asian=0.05%; European, North=70.29%; European, South=28%.
Mutation; HGNC; HGNC Mutation; HGNC; HGNC
A*02
In situ; Lymph node; 
Caucasian.
Cancer cell line
Assay: HTRF
Dmax:  99 %
Assay description: Effects of Compounds on BTK Degradation in OCI-LY10 Cells
Total-BTK Kits (cisbio, 63ADK064PEH) were used for detection. 4X Supplemented Lysis Buffer was prepared using Blocking Reagent Stock Solution (100X) and Lysis Buffer Stock Solution (4X). Premixed Antibody Solutions were prepared using Detection Buffer, Total-BTK d2 Antibody, and Total-BTK Eu Cryptate Antibody.
Well-growing OCI-LY10 cells were harvested into a centrifuge tube and the cell density was adjusted to 4.17×10? cells/mL. The cells were seeded onto 384-well plates (12 μL/well) and cultured overnight in a cell culture incubator. Compounds were added using a nanoliter pipette to a final concentration of 100 nM to 0.098 nM. Each concentration was replicated in duplicate wells. A control was also included. After 24 hours of incubation in a cell culture incubator, cells were lysed by adding 4 μL/well of 4X Supplemented Lysis Buffer. After shaking at room temperature for 40 minutes, 4 μL/well of premixed antibody solutions were added. The cells were allowed to stand at room temperature overnight. Fluorescence was measured at 665 nm/620 nm using an Envision microplate reader. Four-parameter analysis was performed, and a dose-response curve was fitted to calculate Dmax (maximum degradation rate). The results are shown in Table 1.
Reference: BRUTON'S TYROSINE KINASE DEGRADER CONTAINING FUSED-RING OR SPIRO-RING (patent)
Source database: TPDdb
Compound (SMILES): Nc1nccn2c(N3CCC4(CC3)CN(CCOCCCOc3ccc5c(c3)CN(C3CCC(=O)NC3=O)C5=O)C4)nc(-c3ccc(C(=O)Nc4cc(Cl)ccn4)cc3F)c12
Modality: PROteolysis-TArgeting Chimera (PROTAC)
Target protein (POI): BTK (UniProt Q06187)
POI sequence: MAAVILESIFLKRSQQKKKTSPLNFKKRLFLLTVHKLSYYEYDFERGRRGSKKGSIDVEKITCVETVVPEKNPPPERQIPRRGEESSEMEQISIIERFPYPFQVVYDEGPLYVFSPTEELRKRWIHQLKNVIRYNSDLVQKYHPCFWIDGQYLCCSQTAKNAMGCQILENRNGSLKPGSSHRKTKKPLPPTPEEDQILKKPLPPEPAAAPVSTSELKKVVALYDYMPMNANDLQLRKGDEYFILEESNLPWWRARDKNGQEGYIPSNYVTEAEDSIEMYEWYSKHMTRSQAEQLLKQEGKEGGFIVRDSSKAGKYTVSVFAKSTGDPQGVIRHYVVCSTPQSQYYLAEKHLFSTIPELINYHQHNSAGLISRLKYPVSQQNKNAPSTAGLGYGSWEIDPKDLTFLKELGTGQFGVVKYGKWRGQYDVAIKMIKEGSMSEDEFIEEAKVMMNLSHEKLVQLYGVCTKQRPIFIITEYMANGCLLNYLREMRHRFQTQQLLEMCKDVCEAMEYLESKQFLHRDLAARNCLVNDQGVVKVSDFGLSRYVLDDEYTSSVGSKFPVRWSPPEVLMYSKFSSKSDIWAFGVLMWEIYSLGKMPYERFTNSETAEHIAQGLRLYRPHLASEKVYTIMYSCWHEKADERPTFKILLSNILDVMDEES
E3 ligase: CRBN (UniProt Q96SW2)
Ligase sequence: MAGEGDQQDAAHNMGNHLPLLPAESEEEDEMEVEDQDSKEAKKPNIINFDTSLPTSHTYLGADMEEFHGRTLHDDDSCQVIPVLPQVMMILIPGQTLPLQLFHPQEVSMVRNLIQKDRTFAVLAYSNVQEREAQFGTTAEIYAYREEQDFGIEIVKVKAIGRQRFKVLELRTQSDGIQQAKVQILPECVLPSTMSAVQLESLNKCQIFPSKPVSREDQCSYKWWQKYQKRKFHCANLTSWPRWLYSLYDAETLMDRIKKQLREWDENLKDDSLPSNPIDFSYRVAACLPIDDVLRIQLLKIGSAIQRLRCELDIMNKCTSLCCKQCQETEITTKNEIFSLSLCGPMAAYVNPHGYVHETLTVYKACNLNLIGRPSTEHSWFPGYAWTVAQCKICASHIGWKFTATKKDMSPQKFWGLTRSALLPTIPDTEDEISPDKVILCL
Cell line: OCI-Ly10 (Homo sapiens)
Cell line description: African=0.25%; Native American=0.93%; East Asian, North=0.48%; East Asian, South=0%; South Asian=0.05%; European, North=70.29%; European, South=28%.
Mutation; HGNC; HGNC Mutation; HGNC; HGNC
A*02
In situ; Lymph node; 
Caucasian.
Cancer cell line
Assay: HTRF
Dmax:  97 %
Assay description: Effects of Compounds on BTK Degradation in OCI-LY10 Cells
Total-BTK Kits (cisbio, 63ADK064PEH) were used for detection. 4X Supplemented Lysis Buffer was prepared using Blocking Reagent Stock Solution (100X) and Lysis Buffer Stock Solution (4X). Premixed Antibody Solutions were prepared using Detection Buffer, Total-BTK d2 Antibody, and Total-BTK Eu Cryptate Antibody.
Well-growing OCI-LY10 cells were harvested into a centrifuge tube and the cell density was adjusted to 4.17×10? cells/mL. The cells were seeded onto 384-well plates (12 μL/well) and cultured overnight in a cell culture incubator. Compounds were added using a nanoliter pipette to a final concentration of 100 nM to 0.098 nM. Each concentration was replicated in duplicate wells. A control was also included. After 24 hours of incubation in a cell culture incubator, cells were lysed by adding 4 μL/well of 4X Supplemented Lysis Buffer. After shaking at room temperature for 40 minutes, 4 μL/well of premixed antibody solutions were added. The cells were allowed to stand at room temperature overnight. Fluorescence was measured at 665 nm/620 nm using an Envision microplate reader. Four-parameter analysis was performed, and a dose-response curve was fitted to calculate Dmax (maximum degradation rate). The results are shown in Table 1.
Reference: BRUTON'S TYROSINE KINASE DEGRADER CONTAINING FUSED-RING OR SPIRO-RING (patent)
Source database: TPDdb
Compound (SMILES): Nc1nccn2c(N3CCC4(CC3)CN(CCOCCOCCNc3ccc5c(c3)CN(C3CCC(=O)NC3=O)C5=O)C4)nc(-c3ccc(C(=O)Nc4cc(Cl)ccn4)cc3F)c12
Modality: PROteolysis-TArgeting Chimera (PROTAC)
Target protein (POI): BTK (UniProt Q06187)
POI sequence: MAAVILESIFLKRSQQKKKTSPLNFKKRLFLLTVHKLSYYEYDFERGRRGSKKGSIDVEKITCVETVVPEKNPPPERQIPRRGEESSEMEQISIIERFPYPFQVVYDEGPLYVFSPTEELRKRWIHQLKNVIRYNSDLVQKYHPCFWIDGQYLCCSQTAKNAMGCQILENRNGSLKPGSSHRKTKKPLPPTPEEDQILKKPLPPEPAAAPVSTSELKKVVALYDYMPMNANDLQLRKGDEYFILEESNLPWWRARDKNGQEGYIPSNYVTEAEDSIEMYEWYSKHMTRSQAEQLLKQEGKEGGFIVRDSSKAGKYTVSVFAKSTGDPQGVIRHYVVCSTPQSQYYLAEKHLFSTIPELINYHQHNSAGLISRLKYPVSQQNKNAPSTAGLGYGSWEIDPKDLTFLKELGTGQFGVVKYGKWRGQYDVAIKMIKEGSMSEDEFIEEAKVMMNLSHEKLVQLYGVCTKQRPIFIITEYMANGCLLNYLREMRHRFQTQQLLEMCKDVCEAMEYLESKQFLHRDLAARNCLVNDQGVVKVSDFGLSRYVLDDEYTSSVGSKFPVRWSPPEVLMYSKFSSKSDIWAFGVLMWEIYSLGKMPYERFTNSETAEHIAQGLRLYRPHLASEKVYTIMYSCWHEKADERPTFKILLSNILDVMDEES
E3 ligase: CRBN (UniProt Q96SW2)
Ligase sequence: MAGEGDQQDAAHNMGNHLPLLPAESEEEDEMEVEDQDSKEAKKPNIINFDTSLPTSHTYLGADMEEFHGRTLHDDDSCQVIPVLPQVMMILIPGQTLPLQLFHPQEVSMVRNLIQKDRTFAVLAYSNVQEREAQFGTTAEIYAYREEQDFGIEIVKVKAIGRQRFKVLELRTQSDGIQQAKVQILPECVLPSTMSAVQLESLNKCQIFPSKPVSREDQCSYKWWQKYQKRKFHCANLTSWPRWLYSLYDAETLMDRIKKQLREWDENLKDDSLPSNPIDFSYRVAACLPIDDVLRIQLLKIGSAIQRLRCELDIMNKCTSLCCKQCQETEITTKNEIFSLSLCGPMAAYVNPHGYVHETLTVYKACNLNLIGRPSTEHSWFPGYAWTVAQCKICASHIGWKFTATKKDMSPQKFWGLTRSALLPTIPDTEDEISPDKVILCL
Cell line: OCI-Ly10 (Homo sapiens)
Cell line description: African=0.25%; Native American=0.93%; East Asian, North=0.48%; East Asian, South=0%; South Asian=0.05%; European, North=70.29%; European, South=28%.
Mutation; HGNC; HGNC Mutation; HGNC; HGNC
A*02
In situ; Lymph node; 
Caucasian.
Cancer cell line
Assay: HTRF
Dmax:  96 %
Assay description: Effects of Compounds on BTK Degradation in OCI-LY10 Cells
Total-BTK Kits (cisbio, 63ADK064PEH) were used for detection. 4X Supplemented Lysis Buffer was prepared using Blocking Reagent Stock Solution (100X) and Lysis Buffer Stock Solution (4X). Premixed Antibody Solutions were prepared using Detection Buffer, Total-BTK d2 Antibody, and Total-BTK Eu Cryptate Antibody.
Well-growing OCI-LY10 cells were harvested into a centrifuge tube and the cell density was adjusted to 4.17×10? cells/mL. The cells were seeded onto 384-well plates (12 μL/well) and cultured overnight in a cell culture incubator. Compounds were added using a nanoliter pipette to a final concentration of 100 nM to 0.098 nM. Each concentration was replicated in duplicate wells. A control was also included. After 24 hours of incubation in a cell culture incubator, cells were lysed by adding 4 μL/well of 4X Supplemented Lysis Buffer. After shaking at room temperature for 40 minutes, 4 μL/well of premixed antibody solutions were added. The cells were allowed to stand at room temperature overnight. Fluorescence was measured at 665 nm/620 nm using an Envision microplate reader. Four-parameter analysis was performed, and a dose-response curve was fitted to calculate Dmax (maximum degradation rate). The results are shown in Table 1.
Reference: BRUTON'S TYROSINE KINASE DEGRADER CONTAINING FUSED-RING OR SPIRO-RING (patent)
Source database: TPDdb
Compound (SMILES): Nc1nccn2c(N3CCC4(CC3)CN(CCOCCOCCOc3ccc5c(c3)C(=O)N(C3CCC(=O)NC3=O)C5)C4)nc(-c3ccc(C(=O)Nc4cc(Cl)ccn4)cc3F)c12
Modality: PROteolysis-TArgeting Chimera (PROTAC)
Target protein (POI): BTK (UniProt Q06187)
POI sequence: MAAVILESIFLKRSQQKKKTSPLNFKKRLFLLTVHKLSYYEYDFERGRRGSKKGSIDVEKITCVETVVPEKNPPPERQIPRRGEESSEMEQISIIERFPYPFQVVYDEGPLYVFSPTEELRKRWIHQLKNVIRYNSDLVQKYHPCFWIDGQYLCCSQTAKNAMGCQILENRNGSLKPGSSHRKTKKPLPPTPEEDQILKKPLPPEPAAAPVSTSELKKVVALYDYMPMNANDLQLRKGDEYFILEESNLPWWRARDKNGQEGYIPSNYVTEAEDSIEMYEWYSKHMTRSQAEQLLKQEGKEGGFIVRDSSKAGKYTVSVFAKSTGDPQGVIRHYVVCSTPQSQYYLAEKHLFSTIPELINYHQHNSAGLISRLKYPVSQQNKNAPSTAGLGYGSWEIDPKDLTFLKELGTGQFGVVKYGKWRGQYDVAIKMIKEGSMSEDEFIEEAKVMMNLSHEKLVQLYGVCTKQRPIFIITEYMANGCLLNYLREMRHRFQTQQLLEMCKDVCEAMEYLESKQFLHRDLAARNCLVNDQGVVKVSDFGLSRYVLDDEYTSSVGSKFPVRWSPPEVLMYSKFSSKSDIWAFGVLMWEIYSLGKMPYERFTNSETAEHIAQGLRLYRPHLASEKVYTIMYSCWHEKADERPTFKILLSNILDVMDEES
E3 ligase: CRBN (UniProt Q96SW2)
Ligase sequence: MAGEGDQQDAAHNMGNHLPLLPAESEEEDEMEVEDQDSKEAKKPNIINFDTSLPTSHTYLGADMEEFHGRTLHDDDSCQVIPVLPQVMMILIPGQTLPLQLFHPQEVSMVRNLIQKDRTFAVLAYSNVQEREAQFGTTAEIYAYREEQDFGIEIVKVKAIGRQRFKVLELRTQSDGIQQAKVQILPECVLPSTMSAVQLESLNKCQIFPSKPVSREDQCSYKWWQKYQKRKFHCANLTSWPRWLYSLYDAETLMDRIKKQLREWDENLKDDSLPSNPIDFSYRVAACLPIDDVLRIQLLKIGSAIQRLRCELDIMNKCTSLCCKQCQETEITTKNEIFSLSLCGPMAAYVNPHGYVHETLTVYKACNLNLIGRPSTEHSWFPGYAWTVAQCKICASHIGWKFTATKKDMSPQKFWGLTRSALLPTIPDTEDEISPDKVILCL
Cell line: OCI-Ly10 (Homo sapiens)
Cell line description: African=0.25%; Native American=0.93%; East Asian, North=0.48%; East Asian, South=0%; South Asian=0.05%; European, North=70.29%; European, South=28%.
Mutation; HGNC; HGNC Mutation; HGNC; HGNC
A*02
In situ; Lymph node; 
Caucasian.
Cancer cell line
Assay: HTRF
Dmax:  97 %
Assay description: Effects of Compounds on BTK Degradation in OCI-LY10 Cells
Total-BTK Kits (cisbio, 63ADK064PEH) were used for detection. 4X Supplemented Lysis Buffer was prepared using Blocking Reagent Stock Solution (100X) and Lysis Buffer Stock Solution (4X). Premixed Antibody Solutions were prepared using Detection Buffer, Total-BTK d2 Antibody, and Total-BTK Eu Cryptate Antibody.
Well-growing OCI-LY10 cells were harvested into a centrifuge tube and the cell density was adjusted to 4.17×10? cells/mL. The cells were seeded onto 384-well plates (12 μL/well) and cultured overnight in a cell culture incubator. Compounds were added using a nanoliter pipette to a final concentration of 100 nM to 0.098 nM. Each concentration was replicated in duplicate wells. A control was also included. After 24 hours of incubation in a cell culture incubator, cells were lysed by adding 4 μL/well of 4X Supplemented Lysis Buffer. After shaking at room temperature for 40 minutes, 4 μL/well of premixed antibody solutions were added. The cells were allowed to stand at room temperature overnight. Fluorescence was measured at 665 nm/620 nm using an Envision microplate reader. Four-parameter analysis was performed, and a dose-response curve was fitted to calculate Dmax (maximum degradation rate). The results are shown in Table 1.
Reference: BRUTON'S TYROSINE KINASE DEGRADER CONTAINING FUSED-RING OR SPIRO-RING (patent)
Source database: TPDdb
Compound (SMILES): Nc1nccn2c(N3CCC4(CC3)CN(CCOCCOCCOc3ccc5c(c3)CN(C3CCC(=O)NC3=O)C5=O)C4)nc(-c3ccc(C(=O)Nc4cc(Cl)ccn4)cc3F)c12
Modality: PROteolysis-TArgeting Chimera (PROTAC)
Target protein (POI): BTK (UniProt Q06187)
POI sequence: MAAVILESIFLKRSQQKKKTSPLNFKKRLFLLTVHKLSYYEYDFERGRRGSKKGSIDVEKITCVETVVPEKNPPPERQIPRRGEESSEMEQISIIERFPYPFQVVYDEGPLYVFSPTEELRKRWIHQLKNVIRYNSDLVQKYHPCFWIDGQYLCCSQTAKNAMGCQILENRNGSLKPGSSHRKTKKPLPPTPEEDQILKKPLPPEPAAAPVSTSELKKVVALYDYMPMNANDLQLRKGDEYFILEESNLPWWRARDKNGQEGYIPSNYVTEAEDSIEMYEWYSKHMTRSQAEQLLKQEGKEGGFIVRDSSKAGKYTVSVFAKSTGDPQGVIRHYVVCSTPQSQYYLAEKHLFSTIPELINYHQHNSAGLISRLKYPVSQQNKNAPSTAGLGYGSWEIDPKDLTFLKELGTGQFGVVKYGKWRGQYDVAIKMIKEGSMSEDEFIEEAKVMMNLSHEKLVQLYGVCTKQRPIFIITEYMANGCLLNYLREMRHRFQTQQLLEMCKDVCEAMEYLESKQFLHRDLAARNCLVNDQGVVKVSDFGLSRYVLDDEYTSSVGSKFPVRWSPPEVLMYSKFSSKSDIWAFGVLMWEIYSLGKMPYERFTNSETAEHIAQGLRLYRPHLASEKVYTIMYSCWHEKADERPTFKILLSNILDVMDEES
E3 ligase: CRBN (UniProt Q96SW2)
Ligase sequence: MAGEGDQQDAAHNMGNHLPLLPAESEEEDEMEVEDQDSKEAKKPNIINFDTSLPTSHTYLGADMEEFHGRTLHDDDSCQVIPVLPQVMMILIPGQTLPLQLFHPQEVSMVRNLIQKDRTFAVLAYSNVQEREAQFGTTAEIYAYREEQDFGIEIVKVKAIGRQRFKVLELRTQSDGIQQAKVQILPECVLPSTMSAVQLESLNKCQIFPSKPVSREDQCSYKWWQKYQKRKFHCANLTSWPRWLYSLYDAETLMDRIKKQLREWDENLKDDSLPSNPIDFSYRVAACLPIDDVLRIQLLKIGSAIQRLRCELDIMNKCTSLCCKQCQETEITTKNEIFSLSLCGPMAAYVNPHGYVHETLTVYKACNLNLIGRPSTEHSWFPGYAWTVAQCKICASHIGWKFTATKKDMSPQKFWGLTRSALLPTIPDTEDEISPDKVILCL
Cell line: OCI-Ly10 (Homo sapiens)
Cell line description: African=0.25%; Native American=0.93%; East Asian, North=0.48%; East Asian, South=0%; South Asian=0.05%; European, North=70.29%; European, South=28%.
Mutation; HGNC; HGNC Mutation; HGNC; HGNC
A*02
In situ; Lymph node; 
Caucasian.
Cancer cell line
Assay: HTRF
Dmax:  99 %
Assay description: Effects of Compounds on BTK Degradation in OCI-LY10 Cells
Total-BTK Kits (cisbio, 63ADK064PEH) were used for detection. 4X Supplemented Lysis Buffer was prepared using Blocking Reagent Stock Solution (100X) and Lysis Buffer Stock Solution (4X). Premixed Antibody Solutions were prepared using Detection Buffer, Total-BTK d2 Antibody, and Total-BTK Eu Cryptate Antibody.
Well-growing OCI-LY10 cells were harvested into a centrifuge tube and the cell density was adjusted to 4.17×10? cells/mL. The cells were seeded onto 384-well plates (12 μL/well) and cultured overnight in a cell culture incubator. Compounds were added using a nanoliter pipette to a final concentration of 100 nM to 0.098 nM. Each concentration was replicated in duplicate wells. A control was also included. After 24 hours of incubation in a cell culture incubator, cells were lysed by adding 4 μL/well of 4X Supplemented Lysis Buffer. After shaking at room temperature for 40 minutes, 4 μL/well of premixed antibody solutions were added. The cells were allowed to stand at room temperature overnight. Fluorescence was measured at 665 nm/620 nm using an Envision microplate reader. Four-parameter analysis was performed, and a dose-response curve was fitted to calculate Dmax (maximum degradation rate). The results are shown in Table 1.
Reference: BRUTON'S TYROSINE KINASE DEGRADER CONTAINING FUSED-RING OR SPIRO-RING (patent)
Source database: TPDdb
Compound (SMILES): Nc1nccn2c(N3CCC4(CC3)CN(CCOCCOCCOc3cccc5c3C(=O)N(C3CCC(=O)NC3=O)C5)C4)nc(-c3ccc(C(=O)Nc4cc(Cl)ccn4)cc3F)c12
Modality: PROteolysis-TArgeting Chimera (PROTAC)
Target protein (POI): BTK (UniProt Q06187)
POI sequence: MAAVILESIFLKRSQQKKKTSPLNFKKRLFLLTVHKLSYYEYDFERGRRGSKKGSIDVEKITCVETVVPEKNPPPERQIPRRGEESSEMEQISIIERFPYPFQVVYDEGPLYVFSPTEELRKRWIHQLKNVIRYNSDLVQKYHPCFWIDGQYLCCSQTAKNAMGCQILENRNGSLKPGSSHRKTKKPLPPTPEEDQILKKPLPPEPAAAPVSTSELKKVVALYDYMPMNANDLQLRKGDEYFILEESNLPWWRARDKNGQEGYIPSNYVTEAEDSIEMYEWYSKHMTRSQAEQLLKQEGKEGGFIVRDSSKAGKYTVSVFAKSTGDPQGVIRHYVVCSTPQSQYYLAEKHLFSTIPELINYHQHNSAGLISRLKYPVSQQNKNAPSTAGLGYGSWEIDPKDLTFLKELGTGQFGVVKYGKWRGQYDVAIKMIKEGSMSEDEFIEEAKVMMNLSHEKLVQLYGVCTKQRPIFIITEYMANGCLLNYLREMRHRFQTQQLLEMCKDVCEAMEYLESKQFLHRDLAARNCLVNDQGVVKVSDFGLSRYVLDDEYTSSVGSKFPVRWSPPEVLMYSKFSSKSDIWAFGVLMWEIYSLGKMPYERFTNSETAEHIAQGLRLYRPHLASEKVYTIMYSCWHEKADERPTFKILLSNILDVMDEES
E3 ligase: CRBN (UniProt Q96SW2)
Ligase sequence: MAGEGDQQDAAHNMGNHLPLLPAESEEEDEMEVEDQDSKEAKKPNIINFDTSLPTSHTYLGADMEEFHGRTLHDDDSCQVIPVLPQVMMILIPGQTLPLQLFHPQEVSMVRNLIQKDRTFAVLAYSNVQEREAQFGTTAEIYAYREEQDFGIEIVKVKAIGRQRFKVLELRTQSDGIQQAKVQILPECVLPSTMSAVQLESLNKCQIFPSKPVSREDQCSYKWWQKYQKRKFHCANLTSWPRWLYSLYDAETLMDRIKKQLREWDENLKDDSLPSNPIDFSYRVAACLPIDDVLRIQLLKIGSAIQRLRCELDIMNKCTSLCCKQCQETEITTKNEIFSLSLCGPMAAYVNPHGYVHETLTVYKACNLNLIGRPSTEHSWFPGYAWTVAQCKICASHIGWKFTATKKDMSPQKFWGLTRSALLPTIPDTEDEISPDKVILCL
Cell line: OCI-Ly10 (Homo sapiens)
Cell line description: African=0.25%; Native American=0.93%; East Asian, North=0.48%; East Asian, South=0%; South Asian=0.05%; European, North=70.29%; European, South=28%.
Mutation; HGNC; HGNC Mutation; HGNC; HGNC
A*02
In situ; Lymph node; 
Caucasian.
Cancer cell line
Assay: HTRF
Dmax:  91 %
Assay description: Effects of Compounds on BTK Degradation in OCI-LY10 Cells
Total-BTK Kits (cisbio, 63ADK064PEH) were used for detection. 4X Supplemented Lysis Buffer was prepared using Blocking Reagent Stock Solution (100X) and Lysis Buffer Stock Solution (4X). Premixed Antibody Solutions were prepared using Detection Buffer, Total-BTK d2 Antibody, and Total-BTK Eu Cryptate Antibody.
Well-growing OCI-LY10 cells were harvested into a centrifuge tube and the cell density was adjusted to 4.17×10? cells/mL. The cells were seeded onto 384-well plates (12 μL/well) and cultured overnight in a cell culture incubator. Compounds were added using a nanoliter pipette to a final concentration of 100 nM to 0.098 nM. Each concentration was replicated in duplicate wells. A control was also included. After 24 hours of incubation in a cell culture incubator, cells were lysed by adding 4 μL/well of 4X Supplemented Lysis Buffer. After shaking at room temperature for 40 minutes, 4 μL/well of premixed antibody solutions were added. The cells were allowed to stand at room temperature overnight. Fluorescence was measured at 665 nm/620 nm using an Envision microplate reader. Four-parameter analysis was performed, and a dose-response curve was fitted to calculate Dmax (maximum degradation rate). The results are shown in Table 1.
Reference: BRUTON'S TYROSINE KINASE DEGRADER CONTAINING FUSED-RING OR SPIRO-RING (patent)
Source database: TPDdb
Compound (SMILES): Nc1nccn2c(N3CCC4(CC3)CN(C[C@@H]3CCN(c5ccc6c(c5)C(=O)N(C5CCC(=O)NC5=O)C6=O)C3)C4)nc(-c3ccc(C(=O)Nc4cc(Cl)ccn4)cc3F)c12
Modality: PROteolysis-TArgeting Chimera (PROTAC)
Target protein (POI): BTK (UniProt Q06187)
POI sequence: MAAVILESIFLKRSQQKKKTSPLNFKKRLFLLTVHKLSYYEYDFERGRRGSKKGSIDVEKITCVETVVPEKNPPPERQIPRRGEESSEMEQISIIERFPYPFQVVYDEGPLYVFSPTEELRKRWIHQLKNVIRYNSDLVQKYHPCFWIDGQYLCCSQTAKNAMGCQILENRNGSLKPGSSHRKTKKPLPPTPEEDQILKKPLPPEPAAAPVSTSELKKVVALYDYMPMNANDLQLRKGDEYFILEESNLPWWRARDKNGQEGYIPSNYVTEAEDSIEMYEWYSKHMTRSQAEQLLKQEGKEGGFIVRDSSKAGKYTVSVFAKSTGDPQGVIRHYVVCSTPQSQYYLAEKHLFSTIPELINYHQHNSAGLISRLKYPVSQQNKNAPSTAGLGYGSWEIDPKDLTFLKELGTGQFGVVKYGKWRGQYDVAIKMIKEGSMSEDEFIEEAKVMMNLSHEKLVQLYGVCTKQRPIFIITEYMANGCLLNYLREMRHRFQTQQLLEMCKDVCEAMEYLESKQFLHRDLAARNCLVNDQGVVKVSDFGLSRYVLDDEYTSSVGSKFPVRWSPPEVLMYSKFSSKSDIWAFGVLMWEIYSLGKMPYERFTNSETAEHIAQGLRLYRPHLASEKVYTIMYSCWHEKADERPTFKILLSNILDVMDEES
E3 ligase: CRBN (UniProt Q96SW2)
Ligase sequence: MAGEGDQQDAAHNMGNHLPLLPAESEEEDEMEVEDQDSKEAKKPNIINFDTSLPTSHTYLGADMEEFHGRTLHDDDSCQVIPVLPQVMMILIPGQTLPLQLFHPQEVSMVRNLIQKDRTFAVLAYSNVQEREAQFGTTAEIYAYREEQDFGIEIVKVKAIGRQRFKVLELRTQSDGIQQAKVQILPECVLPSTMSAVQLESLNKCQIFPSKPVSREDQCSYKWWQKYQKRKFHCANLTSWPRWLYSLYDAETLMDRIKKQLREWDENLKDDSLPSNPIDFSYRVAACLPIDDVLRIQLLKIGSAIQRLRCELDIMNKCTSLCCKQCQETEITTKNEIFSLSLCGPMAAYVNPHGYVHETLTVYKACNLNLIGRPSTEHSWFPGYAWTVAQCKICASHIGWKFTATKKDMSPQKFWGLTRSALLPTIPDTEDEISPDKVILCL
Cell line: OCI-Ly10 (Homo sapiens)
Cell line description: African=0.25%; Native American=0.93%; East Asian, North=0.48%; East Asian, South=0%; South Asian=0.05%; European, North=70.29%; European, South=28%.
Mutation; HGNC; HGNC Mutation; HGNC; HGNC
A*02
In situ; Lymph node; 
Caucasian.
Cancer cell line
Assay: HTRF
Dmax:  95 %
Assay description: Effects of Compounds on BTK Degradation in OCI-LY10 Cells
Total-BTK Kits (cisbio, 63ADK064PEH) were used for detection. 4X Supplemented Lysis Buffer was prepared using Blocking Reagent Stock Solution (100X) and Lysis Buffer Stock Solution (4X). Premixed Antibody Solutions were prepared using Detection Buffer, Total-BTK d2 Antibody, and Total-BTK Eu Cryptate Antibody.
Well-growing OCI-LY10 cells were harvested into a centrifuge tube and the cell density was adjusted to 4.17×10? cells/mL. The cells were seeded onto 384-well plates (12 μL/well) and cultured overnight in a cell culture incubator. Compounds were added using a nanoliter pipette to a final concentration of 100 nM to 0.098 nM. Each concentration was replicated in duplicate wells. A control was also included. After 24 hours of incubation in a cell culture incubator, cells were lysed by adding 4 μL/well of 4X Supplemented Lysis Buffer. After shaking at room temperature for 40 minutes, 4 μL/well of premixed antibody solutions were added. The cells were allowed to stand at room temperature overnight. Fluorescence was measured at 665 nm/620 nm using an Envision microplate reader. Four-parameter analysis was performed, and a dose-response curve was fitted to calculate Dmax (maximum degradation rate). The results are shown in Table 1.
Reference: BRUTON'S TYROSINE KINASE DEGRADER CONTAINING FUSED-RING OR SPIRO-RING (patent)
Source database: TPDdb
Compound (SMILES): Nc1nccn2c(N3CCC4(CC3)CN(C[C@H]3CCN(c5ccc6c(c5)C(=O)N(C5CCC(=O)NC5=O)C6=O)C3)C4)nc(-c3ccc(C(=O)Nc4cc(Cl)ccn4)cc3F)c12
Modality: PROteolysis-TArgeting Chimera (PROTAC)
Target protein (POI): BTK (UniProt Q06187)
POI sequence: MAAVILESIFLKRSQQKKKTSPLNFKKRLFLLTVHKLSYYEYDFERGRRGSKKGSIDVEKITCVETVVPEKNPPPERQIPRRGEESSEMEQISIIERFPYPFQVVYDEGPLYVFSPTEELRKRWIHQLKNVIRYNSDLVQKYHPCFWIDGQYLCCSQTAKNAMGCQILENRNGSLKPGSSHRKTKKPLPPTPEEDQILKKPLPPEPAAAPVSTSELKKVVALYDYMPMNANDLQLRKGDEYFILEESNLPWWRARDKNGQEGYIPSNYVTEAEDSIEMYEWYSKHMTRSQAEQLLKQEGKEGGFIVRDSSKAGKYTVSVFAKSTGDPQGVIRHYVVCSTPQSQYYLAEKHLFSTIPELINYHQHNSAGLISRLKYPVSQQNKNAPSTAGLGYGSWEIDPKDLTFLKELGTGQFGVVKYGKWRGQYDVAIKMIKEGSMSEDEFIEEAKVMMNLSHEKLVQLYGVCTKQRPIFIITEYMANGCLLNYLREMRHRFQTQQLLEMCKDVCEAMEYLESKQFLHRDLAARNCLVNDQGVVKVSDFGLSRYVLDDEYTSSVGSKFPVRWSPPEVLMYSKFSSKSDIWAFGVLMWEIYSLGKMPYERFTNSETAEHIAQGLRLYRPHLASEKVYTIMYSCWHEKADERPTFKILLSNILDVMDEES
E3 ligase: CRBN (UniProt Q96SW2)
Ligase sequence: MAGEGDQQDAAHNMGNHLPLLPAESEEEDEMEVEDQDSKEAKKPNIINFDTSLPTSHTYLGADMEEFHGRTLHDDDSCQVIPVLPQVMMILIPGQTLPLQLFHPQEVSMVRNLIQKDRTFAVLAYSNVQEREAQFGTTAEIYAYREEQDFGIEIVKVKAIGRQRFKVLELRTQSDGIQQAKVQILPECVLPSTMSAVQLESLNKCQIFPSKPVSREDQCSYKWWQKYQKRKFHCANLTSWPRWLYSLYDAETLMDRIKKQLREWDENLKDDSLPSNPIDFSYRVAACLPIDDVLRIQLLKIGSAIQRLRCELDIMNKCTSLCCKQCQETEITTKNEIFSLSLCGPMAAYVNPHGYVHETLTVYKACNLNLIGRPSTEHSWFPGYAWTVAQCKICASHIGWKFTATKKDMSPQKFWGLTRSALLPTIPDTEDEISPDKVILCL
Cell line: OCI-Ly10 (Homo sapiens)
Cell line description: African=0.25%; Native American=0.93%; East Asian, North=0.48%; East Asian, South=0%; South Asian=0.05%; European, North=70.29%; European, South=28%.
Mutation; HGNC; HGNC Mutation; HGNC; HGNC
A*02
In situ; Lymph node; 
Caucasian.
Cancer cell line
Assay: HTRF
Dmax:  96 %
Assay description: Effects of Compounds on BTK Degradation in OCI-LY10 Cells
Total-BTK Kits (cisbio, 63ADK064PEH) were used for detection. 4X Supplemented Lysis Buffer was prepared using Blocking Reagent Stock Solution (100X) and Lysis Buffer Stock Solution (4X). Premixed Antibody Solutions were prepared using Detection Buffer, Total-BTK d2 Antibody, and Total-BTK Eu Cryptate Antibody.
Well-growing OCI-LY10 cells were harvested into a centrifuge tube and the cell density was adjusted to 4.17×10? cells/mL. The cells were seeded onto 384-well plates (12 μL/well) and cultured overnight in a cell culture incubator. Compounds were added using a nanoliter pipette to a final concentration of 100 nM to 0.098 nM. Each concentration was replicated in duplicate wells. A control was also included. After 24 hours of incubation in a cell culture incubator, cells were lysed by adding 4 μL/well of 4X Supplemented Lysis Buffer. After shaking at room temperature for 40 minutes, 4 μL/well of premixed antibody solutions were added. The cells were allowed to stand at room temperature overnight. Fluorescence was measured at 665 nm/620 nm using an Envision microplate reader. Four-parameter analysis was performed, and a dose-response curve was fitted to calculate Dmax (maximum degradation rate). The results are shown in Table 1.
Reference: BRUTON'S TYROSINE KINASE DEGRADER CONTAINING FUSED-RING OR SPIRO-RING (patent)
Source database: TPDdb
Compound (SMILES): Nc1nccn2c(N3CCC4(CC3)CN(C[C@H]3CCN(c5ccc6c(c5)CN(C5CCC(=O)NC5=O)C6=O)C3)C4)nc(-c3ccc(C(=O)Nc4cc(Cl)ccn4)cc3F)c12
Modality: PROteolysis-TArgeting Chimera (PROTAC)
Target protein (POI): BTK (UniProt Q06187)
POI sequence: MAAVILESIFLKRSQQKKKTSPLNFKKRLFLLTVHKLSYYEYDFERGRRGSKKGSIDVEKITCVETVVPEKNPPPERQIPRRGEESSEMEQISIIERFPYPFQVVYDEGPLYVFSPTEELRKRWIHQLKNVIRYNSDLVQKYHPCFWIDGQYLCCSQTAKNAMGCQILENRNGSLKPGSSHRKTKKPLPPTPEEDQILKKPLPPEPAAAPVSTSELKKVVALYDYMPMNANDLQLRKGDEYFILEESNLPWWRARDKNGQEGYIPSNYVTEAEDSIEMYEWYSKHMTRSQAEQLLKQEGKEGGFIVRDSSKAGKYTVSVFAKSTGDPQGVIRHYVVCSTPQSQYYLAEKHLFSTIPELINYHQHNSAGLISRLKYPVSQQNKNAPSTAGLGYGSWEIDPKDLTFLKELGTGQFGVVKYGKWRGQYDVAIKMIKEGSMSEDEFIEEAKVMMNLSHEKLVQLYGVCTKQRPIFIITEYMANGCLLNYLREMRHRFQTQQLLEMCKDVCEAMEYLESKQFLHRDLAARNCLVNDQGVVKVSDFGLSRYVLDDEYTSSVGSKFPVRWSPPEVLMYSKFSSKSDIWAFGVLMWEIYSLGKMPYERFTNSETAEHIAQGLRLYRPHLASEKVYTIMYSCWHEKADERPTFKILLSNILDVMDEES
E3 ligase: CRBN (UniProt Q96SW2)
Ligase sequence: MAGEGDQQDAAHNMGNHLPLLPAESEEEDEMEVEDQDSKEAKKPNIINFDTSLPTSHTYLGADMEEFHGRTLHDDDSCQVIPVLPQVMMILIPGQTLPLQLFHPQEVSMVRNLIQKDRTFAVLAYSNVQEREAQFGTTAEIYAYREEQDFGIEIVKVKAIGRQRFKVLELRTQSDGIQQAKVQILPECVLPSTMSAVQLESLNKCQIFPSKPVSREDQCSYKWWQKYQKRKFHCANLTSWPRWLYSLYDAETLMDRIKKQLREWDENLKDDSLPSNPIDFSYRVAACLPIDDVLRIQLLKIGSAIQRLRCELDIMNKCTSLCCKQCQETEITTKNEIFSLSLCGPMAAYVNPHGYVHETLTVYKACNLNLIGRPSTEHSWFPGYAWTVAQCKICASHIGWKFTATKKDMSPQKFWGLTRSALLPTIPDTEDEISPDKVILCL
Cell line: OCI-Ly10 (Homo sapiens)
Cell line description: African=0.25%; Native American=0.93%; East Asian, North=0.48%; East Asian, South=0%; South Asian=0.05%; European, North=70.29%; European, South=28%.
Mutation; HGNC; HGNC Mutation; HGNC; HGNC
A*02
In situ; Lymph node; 
Caucasian.
Cancer cell line
Assay: HTRF
Dmax:  96 %
Assay description: Effects of Compounds on BTK Degradation in OCI-LY10 Cells
Total-BTK Kits (cisbio, 63ADK064PEH) were used for detection. 4X Supplemented Lysis Buffer was prepared using Blocking Reagent Stock Solution (100X) and Lysis Buffer Stock Solution (4X). Premixed Antibody Solutions were prepared using Detection Buffer, Total-BTK d2 Antibody, and Total-BTK Eu Cryptate Antibody.
Well-growing OCI-LY10 cells were harvested into a centrifuge tube and the cell density was adjusted to 4.17×10? cells/mL. The cells were seeded onto 384-well plates (12 μL/well) and cultured overnight in a cell culture incubator. Compounds were added using a nanoliter pipette to a final concentration of 100 nM to 0.098 nM. Each concentration was replicated in duplicate wells. A control was also included. After 24 hours of incubation in a cell culture incubator, cells were lysed by adding 4 μL/well of 4X Supplemented Lysis Buffer. After shaking at room temperature for 40 minutes, 4 μL/well of premixed antibody solutions were added. The cells were allowed to stand at room temperature overnight. Fluorescence was measured at 665 nm/620 nm using an Envision microplate reader. Four-parameter analysis was performed, and a dose-response curve was fitted to calculate Dmax (maximum degradation rate). The results are shown in Table 1.
Reference: BRUTON'S TYROSINE KINASE DEGRADER CONTAINING FUSED-RING OR SPIRO-RING (patent)
Source database: TPDdb
Compound (SMILES): Nc1nccn2c(N3CCC4(CC3)CN([C@@H]3CCN(C5CN(c6ccc7c(c6)C(=O)N(C6CCC(=O)NC6=O)C7=O)C5)C3)C4)nc(-c3ccc(C(=O)Nc4cc(Cl)ccn4)cc3F)c12
Modality: PROteolysis-TArgeting Chimera (PROTAC)
Target protein (POI): BTK (UniProt Q06187)
POI sequence: MAAVILESIFLKRSQQKKKTSPLNFKKRLFLLTVHKLSYYEYDFERGRRGSKKGSIDVEKITCVETVVPEKNPPPERQIPRRGEESSEMEQISIIERFPYPFQVVYDEGPLYVFSPTEELRKRWIHQLKNVIRYNSDLVQKYHPCFWIDGQYLCCSQTAKNAMGCQILENRNGSLKPGSSHRKTKKPLPPTPEEDQILKKPLPPEPAAAPVSTSELKKVVALYDYMPMNANDLQLRKGDEYFILEESNLPWWRARDKNGQEGYIPSNYVTEAEDSIEMYEWYSKHMTRSQAEQLLKQEGKEGGFIVRDSSKAGKYTVSVFAKSTGDPQGVIRHYVVCSTPQSQYYLAEKHLFSTIPELINYHQHNSAGLISRLKYPVSQQNKNAPSTAGLGYGSWEIDPKDLTFLKELGTGQFGVVKYGKWRGQYDVAIKMIKEGSMSEDEFIEEAKVMMNLSHEKLVQLYGVCTKQRPIFIITEYMANGCLLNYLREMRHRFQTQQLLEMCKDVCEAMEYLESKQFLHRDLAARNCLVNDQGVVKVSDFGLSRYVLDDEYTSSVGSKFPVRWSPPEVLMYSKFSSKSDIWAFGVLMWEIYSLGKMPYERFTNSETAEHIAQGLRLYRPHLASEKVYTIMYSCWHEKADERPTFKILLSNILDVMDEES
E3 ligase: CRBN (UniProt Q96SW2)
Ligase sequence: MAGEGDQQDAAHNMGNHLPLLPAESEEEDEMEVEDQDSKEAKKPNIINFDTSLPTSHTYLGADMEEFHGRTLHDDDSCQVIPVLPQVMMILIPGQTLPLQLFHPQEVSMVRNLIQKDRTFAVLAYSNVQEREAQFGTTAEIYAYREEQDFGIEIVKVKAIGRQRFKVLELRTQSDGIQQAKVQILPECVLPSTMSAVQLESLNKCQIFPSKPVSREDQCSYKWWQKYQKRKFHCANLTSWPRWLYSLYDAETLMDRIKKQLREWDENLKDDSLPSNPIDFSYRVAACLPIDDVLRIQLLKIGSAIQRLRCELDIMNKCTSLCCKQCQETEITTKNEIFSLSLCGPMAAYVNPHGYVHETLTVYKACNLNLIGRPSTEHSWFPGYAWTVAQCKICASHIGWKFTATKKDMSPQKFWGLTRSALLPTIPDTEDEISPDKVILCL
Cell line: OCI-Ly10 (Homo sapiens)
Cell line description: African=0.25%; Native American=0.93%; East Asian, North=0.48%; East Asian, South=0%; South Asian=0.05%; European, North=70.29%; European, South=28%.
Mutation; HGNC; HGNC Mutation; HGNC; HGNC
A*02
In situ; Lymph node; 
Caucasian.
Cancer cell line
Assay: HTRF
Dmax:  98 %
Assay description: Effects of Compounds on BTK Degradation in OCI-LY10 Cells
Total-BTK Kits (cisbio, 63ADK064PEH) were used for detection. 4X Supplemented Lysis Buffer was prepared using Blocking Reagent Stock Solution (100X) and Lysis Buffer Stock Solution (4X). Premixed Antibody Solutions were prepared using Detection Buffer, Total-BTK d2 Antibody, and Total-BTK Eu Cryptate Antibody.
Well-growing OCI-LY10 cells were harvested into a centrifuge tube and the cell density was adjusted to 4.17×10? cells/mL. The cells were seeded onto 384-well plates (12 μL/well) and cultured overnight in a cell culture incubator. Compounds were added using a nanoliter pipette to a final concentration of 100 nM to 0.098 nM. Each concentration was replicated in duplicate wells. A control was also included. After 24 hours of incubation in a cell culture incubator, cells were lysed by adding 4 μL/well of 4X Supplemented Lysis Buffer. After shaking at room temperature for 40 minutes, 4 μL/well of premixed antibody solutions were added. The cells were allowed to stand at room temperature overnight. Fluorescence was measured at 665 nm/620 nm using an Envision microplate reader. Four-parameter analysis was performed, and a dose-response curve was fitted to calculate Dmax (maximum degradation rate). The results are shown in Table 1.
Reference: BRUTON'S TYROSINE KINASE DEGRADER CONTAINING FUSED-RING OR SPIRO-RING (patent)
Source database: TPDdb
Compound (SMILES): Nc1nccn2c(N3CCC4(CC3)CN([C@H]3CCN(C5CN(c6ccc7c(c6)C(=O)N(C6CCC(=O)NC6=O)C7=O)C5)C3)C4)nc(-c3ccc(C(=O)Nc4cc(Cl)ccn4)cc3F)c12
Modality: PROteolysis-TArgeting Chimera (PROTAC)
Target protein (POI): BTK (UniProt Q06187)
POI sequence: MAAVILESIFLKRSQQKKKTSPLNFKKRLFLLTVHKLSYYEYDFERGRRGSKKGSIDVEKITCVETVVPEKNPPPERQIPRRGEESSEMEQISIIERFPYPFQVVYDEGPLYVFSPTEELRKRWIHQLKNVIRYNSDLVQKYHPCFWIDGQYLCCSQTAKNAMGCQILENRNGSLKPGSSHRKTKKPLPPTPEEDQILKKPLPPEPAAAPVSTSELKKVVALYDYMPMNANDLQLRKGDEYFILEESNLPWWRARDKNGQEGYIPSNYVTEAEDSIEMYEWYSKHMTRSQAEQLLKQEGKEGGFIVRDSSKAGKYTVSVFAKSTGDPQGVIRHYVVCSTPQSQYYLAEKHLFSTIPELINYHQHNSAGLISRLKYPVSQQNKNAPSTAGLGYGSWEIDPKDLTFLKELGTGQFGVVKYGKWRGQYDVAIKMIKEGSMSEDEFIEEAKVMMNLSHEKLVQLYGVCTKQRPIFIITEYMANGCLLNYLREMRHRFQTQQLLEMCKDVCEAMEYLESKQFLHRDLAARNCLVNDQGVVKVSDFGLSRYVLDDEYTSSVGSKFPVRWSPPEVLMYSKFSSKSDIWAFGVLMWEIYSLGKMPYERFTNSETAEHIAQGLRLYRPHLASEKVYTIMYSCWHEKADERPTFKILLSNILDVMDEES
E3 ligase: CRBN (UniProt Q96SW2)
Ligase sequence: MAGEGDQQDAAHNMGNHLPLLPAESEEEDEMEVEDQDSKEAKKPNIINFDTSLPTSHTYLGADMEEFHGRTLHDDDSCQVIPVLPQVMMILIPGQTLPLQLFHPQEVSMVRNLIQKDRTFAVLAYSNVQEREAQFGTTAEIYAYREEQDFGIEIVKVKAIGRQRFKVLELRTQSDGIQQAKVQILPECVLPSTMSAVQLESLNKCQIFPSKPVSREDQCSYKWWQKYQKRKFHCANLTSWPRWLYSLYDAETLMDRIKKQLREWDENLKDDSLPSNPIDFSYRVAACLPIDDVLRIQLLKIGSAIQRLRCELDIMNKCTSLCCKQCQETEITTKNEIFSLSLCGPMAAYVNPHGYVHETLTVYKACNLNLIGRPSTEHSWFPGYAWTVAQCKICASHIGWKFTATKKDMSPQKFWGLTRSALLPTIPDTEDEISPDKVILCL
Cell line: OCI-Ly10 (Homo sapiens)
Cell line description: African=0.25%; Native American=0.93%; East Asian, North=0.48%; East Asian, South=0%; South Asian=0.05%; European, North=70.29%; European, South=28%.
Mutation; HGNC; HGNC Mutation; HGNC; HGNC
A*02
In situ; Lymph node; 
Caucasian.
Cancer cell line
Assay: HTRF
Dmax:  95 %
Assay description: Effects of Compounds on BTK Degradation in OCI-LY10 Cells
Total-BTK Kits (cisbio, 63ADK064PEH) were used for detection. 4X Supplemented Lysis Buffer was prepared using Blocking Reagent Stock Solution (100X) and Lysis Buffer Stock Solution (4X). Premixed Antibody Solutions were prepared using Detection Buffer, Total-BTK d2 Antibody, and Total-BTK Eu Cryptate Antibody.
Well-growing OCI-LY10 cells were harvested into a centrifuge tube and the cell density was adjusted to 4.17×10? cells/mL. The cells were seeded onto 384-well plates (12 μL/well) and cultured overnight in a cell culture incubator. Compounds were added using a nanoliter pipette to a final concentration of 100 nM to 0.098 nM. Each concentration was replicated in duplicate wells. A control was also included. After 24 hours of incubation in a cell culture incubator, cells were lysed by adding 4 μL/well of 4X Supplemented Lysis Buffer. After shaking at room temperature for 40 minutes, 4 μL/well of premixed antibody solutions were added. The cells were allowed to stand at room temperature overnight. Fluorescence was measured at 665 nm/620 nm using an Envision microplate reader. Four-parameter analysis was performed, and a dose-response curve was fitted to calculate Dmax (maximum degradation rate). The results are shown in Table 1.
Reference: BRUTON'S TYROSINE KINASE DEGRADER CONTAINING FUSED-RING OR SPIRO-RING (patent)
Source database: TPDdb
Compound (SMILES): Nc1nccn2c(N3CCC4(CCN(CCOCCOCCOc5ccc6c(c5)CN(C5CCC(=O)NC5=O)C6=O)C4=O)CC3)nc(-c3ccc(C(=O)Nc4cc(Cl)ccn4)cc3F)c12
Modality: PROteolysis-TArgeting Chimera (PROTAC)
Target protein (POI): BTK (UniProt Q06187)
POI sequence: MAAVILESIFLKRSQQKKKTSPLNFKKRLFLLTVHKLSYYEYDFERGRRGSKKGSIDVEKITCVETVVPEKNPPPERQIPRRGEESSEMEQISIIERFPYPFQVVYDEGPLYVFSPTEELRKRWIHQLKNVIRYNSDLVQKYHPCFWIDGQYLCCSQTAKNAMGCQILENRNGSLKPGSSHRKTKKPLPPTPEEDQILKKPLPPEPAAAPVSTSELKKVVALYDYMPMNANDLQLRKGDEYFILEESNLPWWRARDKNGQEGYIPSNYVTEAEDSIEMYEWYSKHMTRSQAEQLLKQEGKEGGFIVRDSSKAGKYTVSVFAKSTGDPQGVIRHYVVCSTPQSQYYLAEKHLFSTIPELINYHQHNSAGLISRLKYPVSQQNKNAPSTAGLGYGSWEIDPKDLTFLKELGTGQFGVVKYGKWRGQYDVAIKMIKEGSMSEDEFIEEAKVMMNLSHEKLVQLYGVCTKQRPIFIITEYMANGCLLNYLREMRHRFQTQQLLEMCKDVCEAMEYLESKQFLHRDLAARNCLVNDQGVVKVSDFGLSRYVLDDEYTSSVGSKFPVRWSPPEVLMYSKFSSKSDIWAFGVLMWEIYSLGKMPYERFTNSETAEHIAQGLRLYRPHLASEKVYTIMYSCWHEKADERPTFKILLSNILDVMDEES
E3 ligase: CRBN (UniProt Q96SW2)
Ligase sequence: MAGEGDQQDAAHNMGNHLPLLPAESEEEDEMEVEDQDSKEAKKPNIINFDTSLPTSHTYLGADMEEFHGRTLHDDDSCQVIPVLPQVMMILIPGQTLPLQLFHPQEVSMVRNLIQKDRTFAVLAYSNVQEREAQFGTTAEIYAYREEQDFGIEIVKVKAIGRQRFKVLELRTQSDGIQQAKVQILPECVLPSTMSAVQLESLNKCQIFPSKPVSREDQCSYKWWQKYQKRKFHCANLTSWPRWLYSLYDAETLMDRIKKQLREWDENLKDDSLPSNPIDFSYRVAACLPIDDVLRIQLLKIGSAIQRLRCELDIMNKCTSLCCKQCQETEITTKNEIFSLSLCGPMAAYVNPHGYVHETLTVYKACNLNLIGRPSTEHSWFPGYAWTVAQCKICASHIGWKFTATKKDMSPQKFWGLTRSALLPTIPDTEDEISPDKVILCL
Cell line: OCI-Ly10 (Homo sapiens)
Cell line description: African=0.25%; Native American=0.93%; East Asian, North=0.48%; East Asian, South=0%; South Asian=0.05%; European, North=70.29%; European, South=28%.
Mutation; HGNC; HGNC Mutation; HGNC; HGNC
A*02
In situ; Lymph node; 
Caucasian.
Cancer cell line
Assay: HTRF
Dmax:  98 %
Assay description: Effects of Compounds on BTK Degradation in OCI-LY10 Cells
Total-BTK Kits (cisbio, 63ADK064PEH) were used for detection. 4X Supplemented Lysis Buffer was prepared using Blocking Reagent Stock Solution (100X) and Lysis Buffer Stock Solution (4X). Premixed Antibody Solutions were prepared using Detection Buffer, Total-BTK d2 Antibody, and Total-BTK Eu Cryptate Antibody.
Well-growing OCI-LY10 cells were harvested into a centrifuge tube and the cell density was adjusted to 4.17×10? cells/mL. The cells were seeded onto 384-well plates (12 μL/well) and cultured overnight in a cell culture incubator. Compounds were added using a nanoliter pipette to a final concentration of 100 nM to 0.098 nM. Each concentration was replicated in duplicate wells. A control was also included. After 24 hours of incubation in a cell culture incubator, cells were lysed by adding 4 μL/well of 4X Supplemented Lysis Buffer. After shaking at room temperature for 40 minutes, 4 μL/well of premixed antibody solutions were added. The cells were allowed to stand at room temperature overnight. Fluorescence was measured at 665 nm/620 nm using an Envision microplate reader. Four-parameter analysis was performed, and a dose-response curve was fitted to calculate Dmax (maximum degradation rate). The results are shown in Table 1.
Reference: BRUTON'S TYROSINE KINASE DEGRADER CONTAINING FUSED-RING OR SPIRO-RING (patent)
Source database: TPDdb
Compound (SMILES): Nc1ncnc2c1c(-c1ccc(Oc3ccccc3)cc1)nn2C1CC2CN(C3CN(C4CCN(c5ccc(C(=O)NC6CCC(=O)NC6=O)nc5)C4)C3)CC2C1
Modality: PROteolysis-TArgeting Chimera (PROTAC)
Target protein (POI): IKZF1 (UniProt Q13422)
POI sequence: MDADEGQDMSQVSGKESPPVSDTPDEGDEPMPIPEDLSTTSGGQQSSKSDRVVASNVKVETQSDEENGRACEMNGEECAEDLRMLDASGEKMNGSHRDQGSSALSGVGGIRLPNGKLKCDICGIICIGPNVLMVHKRSHTGERPFQCNQCGASFTQKGNLLRHIKLHSGEKPFKCHLCNYACRRRDALTGHLRTHSVGKPHKCGYCGRSYKQRSSLEEHKERCHNYLESMGLPGTLYPVIKEETNHSEMAEDLCKIGSERSLVLDRLASNVAKRKSSMPQKFLGDKGLSDTPYDSSASYEKENEMMKSHVMDQAINNAINYLGAESLRPLVQTPPGGSEVVPVISPMYQLHKPLAEGTPRSNHSAQDSAVENLLLLSKAKLVPSEREASPSNSCQDSTDTESNNEEQRSGLIYLTNHIAPHARNGLSLKEEHRAYDLLRAASENSQDALRVVSTSGEQMKVYKCEHCRVLFLDHVMYTIHMGCHGFRDPFECNMCGYHSQDRYEFSSHITRGEHRFHMS
E3 ligase: CRBN (UniProt Q96SW2)
Ligase sequence: MAGEGDQQDAAHNMGNHLPLLPAESEEEDEMEVEDQDSKEAKKPNIINFDTSLPTSHTYLGADMEEFHGRTLHDDDSCQVIPVLPQVMMILIPGQTLPLQLFHPQEVSMVRNLIQKDRTFAVLAYSNVQEREAQFGTTAEIYAYREEQDFGIEIVKVKAIGRQRFKVLELRTQSDGIQQAKVQILPECVLPSTMSAVQLESLNKCQIFPSKPVSREDQCSYKWWQKYQKRKFHCANLTSWPRWLYSLYDAETLMDRIKKQLREWDENLKDDSLPSNPIDFSYRVAACLPIDDVLRIQLLKIGSAIQRLRCELDIMNKCTSLCCKQCQETEITTKNEIFSLSLCGPMAAYVNPHGYVHETLTVYKACNLNLIGRPSTEHSWFPGYAWTVAQCKICASHIGWKFTATKKDMSPQKFWGLTRSALLPTIPDTEDEISPDKVILCL
Cell line: Mino (Homo sapiens)
Cell line description: African=0.85%; Native American=0%; East Asian, North=1.5%; East Asian, South=0%; South Asian=6.1%; European, North=39.56%; European, South=51.98%.
Mutation; HGNC; HGNC Mutation; HGNC; HGNC Mutation; HGNC; HGNC
A*11 A*11,68; B*35,39; C*04,12. A*11
In situ; Peripheral blood; 
Caucasian.
Cancer cell line
Assay: Western Blot
DC50: > 1000 nM
Assay description: The IKZF1 protein is one of the target protein substrates for the degradation of amine compounds. The IMiD activity of the compound is judged by the IKZF1 protein degradation experiment. The Mino cell samples degraded by the above BTK protein were detected, and the IKZF1 (CST, Cat. 14859S) protein was simultaneously detected by Western blot method. Calculate and obtain the degradation rate of the IKZF1 protein of the administration group relative to the vehicle group IKZF1 protein according to the following formula:

IKZF1% = (1 - (IKZF1 administration/β-actin)/(IKZF1 vehicle/β-actin)) × 100%

IKZF1 administration is the expression level of IKZF1 in Mino cells at different doses of the compound, and IKZF1 vehicle is the expression level of IKZF1 in Mino cells in the control group. The maximum concentration of the compound in this experimental system is 1000 nM. Dmax is the maximum degradation value in the experimental concentration system, and use Graphpad 5.0 software according to the formula Y = 100/(1 + 10^((LogIC50 - X)*HillSlope)) to fit DC50 values.
Reference: PROTAC COMPOUND, PHARMACEUTICAL COMPOSITION COMPRISING SAME, PREPARATION METHOD THEREFOR, AND USE THEREOF (patent)
Source database: TPDdb
Compound (SMILES): Nc1ncnc2c1c(-c1ccc(Oc3ccccc3)cc1)nn2C1CC2CN(C3CN(C4CCN(c5ccc(C(=O)NC6CCC(=O)NC6=O)nc5)C4)C3)CC2C1
Modality: PROteolysis-TArgeting Chimera (PROTAC)
Target protein (POI): IKZF1 (UniProt Q13422)
POI sequence: MDADEGQDMSQVSGKESPPVSDTPDEGDEPMPIPEDLSTTSGGQQSSKSDRVVASNVKVETQSDEENGRACEMNGEECAEDLRMLDASGEKMNGSHRDQGSSALSGVGGIRLPNGKLKCDICGIICIGPNVLMVHKRSHTGERPFQCNQCGASFTQKGNLLRHIKLHSGEKPFKCHLCNYACRRRDALTGHLRTHSVGKPHKCGYCGRSYKQRSSLEEHKERCHNYLESMGLPGTLYPVIKEETNHSEMAEDLCKIGSERSLVLDRLASNVAKRKSSMPQKFLGDKGLSDTPYDSSASYEKENEMMKSHVMDQAINNAINYLGAESLRPLVQTPPGGSEVVPVISPMYQLHKPLAEGTPRSNHSAQDSAVENLLLLSKAKLVPSEREASPSNSCQDSTDTESNNEEQRSGLIYLTNHIAPHARNGLSLKEEHRAYDLLRAASENSQDALRVVSTSGEQMKVYKCEHCRVLFLDHVMYTIHMGCHGFRDPFECNMCGYHSQDRYEFSSHITRGEHRFHMS
E3 ligase: CRBN (UniProt Q96SW2)
Ligase sequence: MAGEGDQQDAAHNMGNHLPLLPAESEEEDEMEVEDQDSKEAKKPNIINFDTSLPTSHTYLGADMEEFHGRTLHDDDSCQVIPVLPQVMMILIPGQTLPLQLFHPQEVSMVRNLIQKDRTFAVLAYSNVQEREAQFGTTAEIYAYREEQDFGIEIVKVKAIGRQRFKVLELRTQSDGIQQAKVQILPECVLPSTMSAVQLESLNKCQIFPSKPVSREDQCSYKWWQKYQKRKFHCANLTSWPRWLYSLYDAETLMDRIKKQLREWDENLKDDSLPSNPIDFSYRVAACLPIDDVLRIQLLKIGSAIQRLRCELDIMNKCTSLCCKQCQETEITTKNEIFSLSLCGPMAAYVNPHGYVHETLTVYKACNLNLIGRPSTEHSWFPGYAWTVAQCKICASHIGWKFTATKKDMSPQKFWGLTRSALLPTIPDTEDEISPDKVILCL
Cell line: Mino (Homo sapiens)
Cell line description: African=0.85%; Native American=0%; East Asian, North=1.5%; East Asian, South=0%; South Asian=6.1%; European, North=39.56%; European, South=51.98%.
Mutation; HGNC; HGNC Mutation; HGNC; HGNC Mutation; HGNC; HGNC
A*11 A*11,68; B*35,39; C*04,12. A*11
In situ; Peripheral blood; 
Caucasian.
Cancer cell line
Assay: Western Blot
Dmax:  40.24 %
Assay description: The IKZF1 protein is one of the target protein substrates for the degradation of amine compounds. The IMiD activity of the compound is judged by the IKZF1 protein degradation experiment. The Mino cell samples degraded by the above BTK protein were detected, and the IKZF1 (CST, Cat. 14859S) protein was simultaneously detected by Western blot method. Calculate and obtain the degradation rate of the IKZF1 protein of the administration group relative to the vehicle group IKZF1 protein according to the following formula:

IKZF1% = (1 - (IKZF1 administration/β-actin)/(IKZF1 vehicle/β-actin)) × 100%

IKZF1 administration is the expression level of IKZF1 in Mino cells at different doses of the compound, and IKZF1 vehicle is the expression level of IKZF1 in Mino cells in the control group. The maximum concentration of the compound in this experimental system is 1000 nM. Dmax is the maximum degradation value in the experimental concentration system, and use Graphpad 5.0 software according to the formula Y = 100/(1 + 10^((LogIC50 - X)*HillSlope)) to fit DC50 values.
Reference: PROTAC COMPOUND, PHARMACEUTICAL COMPOSITION COMPRISING SAME, PREPARATION METHOD THEREFOR, AND USE THEREOF (patent)
Source database: TPDdb
Compound (SMILES): Nc1ncnc2c1c(-c1ccc(Oc3ccccc3)cc1)nn2C1CC2CN(C3CN(C4CCN(c5ccc(NC6CCC(=O)NC6=O)cc5F)C4)C3)CC2C1
Modality: PROteolysis-TArgeting Chimera (PROTAC)
Target protein (POI): IKZF1 (UniProt Q13422)
POI sequence: MDADEGQDMSQVSGKESPPVSDTPDEGDEPMPIPEDLSTTSGGQQSSKSDRVVASNVKVETQSDEENGRACEMNGEECAEDLRMLDASGEKMNGSHRDQGSSALSGVGGIRLPNGKLKCDICGIICIGPNVLMVHKRSHTGERPFQCNQCGASFTQKGNLLRHIKLHSGEKPFKCHLCNYACRRRDALTGHLRTHSVGKPHKCGYCGRSYKQRSSLEEHKERCHNYLESMGLPGTLYPVIKEETNHSEMAEDLCKIGSERSLVLDRLASNVAKRKSSMPQKFLGDKGLSDTPYDSSASYEKENEMMKSHVMDQAINNAINYLGAESLRPLVQTPPGGSEVVPVISPMYQLHKPLAEGTPRSNHSAQDSAVENLLLLSKAKLVPSEREASPSNSCQDSTDTESNNEEQRSGLIYLTNHIAPHARNGLSLKEEHRAYDLLRAASENSQDALRVVSTSGEQMKVYKCEHCRVLFLDHVMYTIHMGCHGFRDPFECNMCGYHSQDRYEFSSHITRGEHRFHMS
E3 ligase: CRBN (UniProt Q96SW2)
Ligase sequence: MAGEGDQQDAAHNMGNHLPLLPAESEEEDEMEVEDQDSKEAKKPNIINFDTSLPTSHTYLGADMEEFHGRTLHDDDSCQVIPVLPQVMMILIPGQTLPLQLFHPQEVSMVRNLIQKDRTFAVLAYSNVQEREAQFGTTAEIYAYREEQDFGIEIVKVKAIGRQRFKVLELRTQSDGIQQAKVQILPECVLPSTMSAVQLESLNKCQIFPSKPVSREDQCSYKWWQKYQKRKFHCANLTSWPRWLYSLYDAETLMDRIKKQLREWDENLKDDSLPSNPIDFSYRVAACLPIDDVLRIQLLKIGSAIQRLRCELDIMNKCTSLCCKQCQETEITTKNEIFSLSLCGPMAAYVNPHGYVHETLTVYKACNLNLIGRPSTEHSWFPGYAWTVAQCKICASHIGWKFTATKKDMSPQKFWGLTRSALLPTIPDTEDEISPDKVILCL
Cell line: Mino (Homo sapiens)
Cell line description: African=0.85%; Native American=0%; East Asian, North=1.5%; East Asian, South=0%; South Asian=6.1%; European, North=39.56%; European, South=51.98%.
Mutation; HGNC; HGNC Mutation; HGNC; HGNC Mutation; HGNC; HGNC
A*11 A*11,68; B*35,39; C*04,12. A*11
In situ; Peripheral blood; 
Caucasian.
Cancer cell line
Assay: Western Blot
DC50: < 10 nM
Assay description: The IKZF1 protein is one of the target protein substrates for the degradation of amine compounds. The IMiD activity of the compound is judged by the IKZF1 protein degradation experiment. The Mino cell samples degraded by the above BTK protein were detected, and the IKZF1 (CST, Cat. 14859S) protein was simultaneously detected by Western blot method. Calculate and obtain the degradation rate of the IKZF1 protein of the administration group relative to the vehicle group IKZF1 protein according to the following formula:

IKZF1% = (1 - (IKZF1 administration/β-actin)/(IKZF1 vehicle/β-actin)) × 100%

IKZF1 administration is the expression level of IKZF1 in Mino cells at different doses of the compound, and IKZF1 vehicle is the expression level of IKZF1 in Mino cells in the control group. The maximum concentration of the compound in this experimental system is 1000 nM. Dmax is the maximum degradation value in the experimental concentration system, and use Graphpad 5.0 software according to the formula Y = 100/(1 + 10^((LogIC50 - X)*HillSlope)) to fit DC50 values.
Reference: PROTAC COMPOUND, PHARMACEUTICAL COMPOSITION COMPRISING SAME, PREPARATION METHOD THEREFOR, AND USE THEREOF (patent)
Source database: TPDdb