SGC-AAK1-1

SGC-AAK1-1 A dual inhibitor of AAK1 and BMP2K/BIKE

This probe is available from Tocris, Cayman Chemical and Sigma.

Its negative control (SGC-AAK1-1N) is available for purchase from Tocris and Sigma.

  • overview
  • properties
  • references
overview
Probe Negative control

 

SGC-AAK1-1

 

SGC-AAK1-1N

AAK1 (adaptor protein 2-associated kinase) and BMP2K/BIKE (BMP-2 inducible kinase) comprise half of the numb-associated kinase (NAK) family, which also includes cyclin G associated kinase (GAK) and STK16/MPSK1 (serine/threonine kinase 16/myristoylated and palmitoylated serine/threonine kinase 1).1

AAK1 is a 104 kDa serine/threonine kinase with broad tissue expression. Within the cell AAK1 localizes to the cell membrane and cytoplasm.2 AAK1 is involved in clathrin-mediated endocytosis (CME), both by direct binding to clathrin and by phosphorylation of the medium subunit of AP-2 (adaptor protein 2).3-5 In this manner, AAK1 has been identified as a negative regulator of Wnt signaling via mediation of LRP6 internalization. Studies have implicated multiple roles for AAK1 in influencing Notch signaling, including priming and redistribution of Numb as well as Notch activation.6-7

Relatively less is known about the highly understudied kinase BIKE. BIKE is broadly expressed, and in the cell, it localizes to nuclear speckles.2 BIKE was originally identified as its expression was observed to increase upon bone morphogenic protein (BMP-2)-induced differentiation of a prechondroblastic cell line.8 The same study provided evidence for BIKE having an important regulatory role in attenuating the program of osteoblast differentiation. Proteomic studies identified BIKE as a clathrin vesicle-associated protein and have also identified interaction between BIKE and Numb.9-10

NAK family domain structures

Location of AAK1 and BIKE on kinome tree

Snapshot of crystal structure of acylaminoindazole bound to BIKE

SGC-AAK1-1 is a chemical probe for AAK1 and BIKE that potently targets the ATP-binding site (AAK1 Ki =  9.1 nM; BIKE Ki = 17 nM). Regarding kinase selectivity, only three kinases were observed to bind SGC-AAK1-1 within 30-fold of the KD of AAK1 in a KINOMEscan assay (DiscoverX) at 1 μM followed by KD determinations: RIOK1 (KD = 72 nM), RIOK3 (KD = 290 nM), and PIP5K1C (KD = 260 nM). In a live cell NanoBRET assay (Promega) SGC-AAK1-1 has potency for ectopically expressed full-length AAK1- and BIKE-Nluc fusion proteins (AAK1 IC50 = 230 nM; BIKE IC50 = 1.5 μM).

A chemically related negative control compound, SGC-AAK1-1N, is provided.

properties
Probe Negative control

 

SGC-AAK1-1

 

SGC-AAK1-1N

Physical and chemical properties for BAY-876
Molecular weight427.52
Molecular formulaC21H25N5O3S
IUPAC nameN-(6-(3-((N,N-diethylsulfamoyl)amino)phenyl)-1H-indazol-3-yl)cyclopropanecarboxamide
MollogP4.185
PSA89.47
No. of chiral centres0
No. of rotatable bonds9
No. of hydrogen bond acceptors8
No. of hydrogen bond donors3
Storage-20 °C as DMSO stock
DissolutionSoluble in DMSO at least up to 10 mM



 

Physical and chemical properties for BAY-588 (Negative Control)
Molecular weight398.48
Molecular formulaC20H22N4O3S
IUPAC nameN-(6-(3-(cyclopropanesulfonamido)phenyl)-1H-indazol-3-yl)isobutyramide
MollogP3.381
PSA87.62
No. of chiral centres0
No. of rotatable bonds7
No. of hydrogen bond acceptors7
No. of hydrogen bond donors3
Storage-20 °C as DMSO stock
DissolutionSoluble in DMSO at least up to 10 mM

SMILES:
SGC-AAK1-1: O=C(C1CC1)NC2=NNC3=C2C=CC(C4=CC(NS(N(CC)CC)(=O)=O)=CC=C4)=C3
SGC-AAK1-1N: O=C(C(C)C)NC1=NNC2=C1C=CC(C3=CC(NS(C4CC4)(=O)=O)=CC=C3)=C2

 

InChI:
SGC-AAK1-1: InChI=1S/C21H25N5O3S/c1-3-26(4-2)30(28,29)25-17-7-5-6-15(12-17)16-10-11-18-19(13-16)23-24-20(18)22-21(27)14-8-9-14/h5-7,10-14,25H,3-4,8-9H2,1-2H3,(H2,22,23,24,27)

SGC-AAK1-1N: InChI=1S/C20H22N4O3S/c1-12(2)20(25)21-19-17-9-6-14(11-18(17)22-23-19)13-4-3-5-15(10-13)24-28(26,27)16-7-8-16/h3-6,9-12,16,24H,7-8H2,1-2H3,(H2,21,22,23,25)

InChIKey:
SGC-AAK1-1: UCBIQZUJJSVQHL-UHFFFAOYSA-N
SGC-AAK1-1N: RAIAORGFMNXPOV-UHFFFAOYSA-N

selectivity profile
in vitro potency
cell based assay data
references
  1. Sorrell, F. J.; Szklarz, M.; Abdul Azeez, K. R.; Elkins, J. M.; Knapp, S., Family-wide Structural Analysis of Human Numb-Associated Protein Kinases. Structure 2016, 24 (3), 401-11.
  2. Thul, P. J.; Akesson, L.; Wiking, M.; Mahdessian, D.; Geladaki, A.; Ait Blal, H.; Alm, T.; Asplund, A.; Bjork, L.; Breckels, L. M.; Backstrom, A.; Danielsson, F.; Fagerberg, L.; Fall, J.; Gatto, L.; Gnann, C.; Hober, S.; Hjelmare, M.; Johansson, F.; Lee, S.; Lindskog, C.; Mulder, J.; Mulvey, C. M.; Nilsson, P.; Oksvold, P.; Rockberg, J.; Schutten, R.; Schwenk, J. M.; Sivertsson, A.; Sjostedt, E.; Skogs, M.; Stadler, C.; Sullivan, D. P.; Tegel, H.; Winsnes, C.; Zhang, C.; Zwahlen, M.; Mardinoglu, A.; Ponten, F.; von Feilitzen, K.; Lilley, K. S.; Uhlen, M.; Lundberg, E., A subcellular map of the human proteome. Science 2017, 356 (6340).
  3. Conner, S. D.; Schmid, S. L., Differential requirements for AP-2 in clathrin-mediated endocytosis. J Cell Biol 2003, 162 (5), 773-9.
  4. Henderson, D. M.; Conner, S. D., A novel AAK1 splice variant functions at multiple steps of the endocytic pathway. Mol Biol Cell 2007, 18 (7), 2698-706.
  5. Neveu, G.; Ziv-Av, A.; Barouch-Bentov, R.; Berkerman, E.; Mulholland, J.; Einav, S., AP-2-associated protein kinase 1 and cyclin G-associated kinase regulate hepatitis C virus entry and are potential drug targets. J Virol 2015, 89 (8), 4387-404.
  6. Gupta-Rossi, N.; Ortica, S.; Meas-Yedid, V.; Heuss, S.; Moretti, J.; Olivo-Marin, J. C.; Israel, A., The adaptor-associated kinase 1, AAK1, is a positive regulator of the Notch pathway. J Biol Chem 2011, 286 (21), 18720-30.
  7. Sorensen, E. B.; Conner, S. D., AAK1 regulates Numb function at an early step in clathrin-mediated endocytosis. Traffic 2008, 9 (10), 1791-800.
  8. Kearns, A. E.; Donohue, M. M.; Sanyal, B.; Demay, M. B., Cloning and characterization of a novel protein kinase that impairs osteoblast differentiation in vitro. J Biol Chem 2001, 276 (45), 42213-8.
  9. Borner, G. H.; Antrobus, R.; Hirst, J.; Bhumbra, G. S.; Kozik, P.; Jackson, L. P.; Sahlender, D. A.; Robinson, M. S., Multivariate proteomic profiling identifies novel accessory proteins of coated vesicles. J Cell Biol 2012, 197 (1), 141-60.
  10. Krieger, J. R.; Taylor, P.; Gajadhar, A. S.; Guha, A.; Moran, M. F.; McGlade, C. J., Identification and selected reaction monitoring (SRM) quantification of endocytosis factors associated with Numb. Mol Cell Proteomics 2013, 12 (2), 499-514.
pk properties
co-crystal structures
synthetic schemes
materials and methods
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