Cover Page The handle http://hdl.handle.net/1887/23082 holds various files of this Leiden University dissertation. Author: Hoogendoorn, Sascha Title: A chemical biology approach for targeting of ligand-drug conjugates Issue Date: 2014-01-23 A Chemical Biology Approach for Targeting of Ligand-Drug Conjugates PROEFSCHRIFT ter verkrijging van de graad van Doctor aan de Universiteit Leiden, op gezag van Rector Magnificus prof. mr. dr. C. J. J. M. Stolker, volgens besluit van het College voor Promoties te verdedigen op donderdag 23 januari 2014 klokke 11.15 uur door Sascha Hoogendoorn Geboren te Amsterdam in 1984 Promotiecommissie Promotores: Prof. dr. H. S. Overkleeft Prof. dr. G. A. van der Marel Co-promotor: Dr. R. G. Boot Overige leden: Prof. dr. J. J. Neefjes Prof. dr. J. Brouwer Prof. dr. J. Lugtenburg Dr. K. M. Bonger Dr. M. van der Stelt Typeset with LATEX Printed by Ridderprint B. V., Ridderkerk Table of Contents List of Abbreviations iv 1 General introduction 1 2 Targeted pH-dependent fluorescent activity-based cathepsin probes 17 3 Synthesis of pH-activatable red fluorescent BODIPY dyes with distinct functionalities 37 4 Targeted delivery of Heat shock protein 70 by sortase-mediated ligation with a synthetic mannose receptor-binding ligand 55 5 Multivalent MPR ligand for endolysosomal targeting of an activity-based cathepsin probe 71 6 Structure-activity relationship studies towards functionalization of dihydropyridine FSHR agonists 87 7 Small-molecule fluorescent agonists for the follicle-stimulating hormone receptor 123 8 Small-molecule FSHR agonists as targeting agents for BNCT 147 9 From LMW to HMW ligands: fluorescently tagged FSH by sortase-mediated ligation 171 10 Summary, Work in Progress and Future Prospects 187 Samenvatting 207 List of Publications 210 Curriculum Vitae 211 iii List of Abbreviations ab ABP Ac ACN AcOH Ahx aq ar ATP BAIB BDP BMP BNCT Boc Boc2 O BODIPY BSA BSH Bu calcd. cAMP cat. CBB CBE CD CD-MPR CHAPS CHO CI-MPR CPZ CRD CRE CTLD CV antibody activity-based probe acetyl acetonitrile acetic acid aminohexanoic acid aqueous aromatic adenosine triphosphate bis(acetoxy)iodobenzene BODIPY bis(monoacylglycero)phosphate boron neutron capture therapy tert-butoxycarbonyl tert-butoxycarbonic anhydride 4,4-difluoro-4-bora-3a,4adiaza-s-indacene bovine serum albumin closo-borane mercaptoundecahydrododecaborate butyl calculated cyclic adenosine monophosphate catalytic coomassie brilliant blue conditurol beta epoxide circular dichroism cation-dependent mannose-6-phosphate receptor 3-[(3-cholamidopropyl)dimethylammonio]-1propanesulfonate chinese hamster ovary (cells) cation-independent mannose-6-phosphate receptor chlorpromazine carbohydrate recognition domain cAMP response element C-type lectin-like domain column volume Cy d δ DBU DC DC cyanine dye doublet chemical shift 1,8-diazabicyclo[5.4.0]undec-7-ene dendritic cell dual color molecular weight marker SDS-PAGE DC-SIGN dendritic cell-specific intercellular adhesion molecule-3-grabbing non-integrin DCE dichloroethane DCM dichloromethane dd double doublet DDQ 2,3-dichloro-5,6-dicyano-1,4-benzoquinone DHP dihydropyridine DIAD diisopropyl azadicarboxylate DiPEA N,N-diisopropylethylamine DMAP 4-(N,N-dimethylamino)-pyridine DMEM Dulbecco’s modified eagle’s medium DMF N,N-dimethylformamide DMSO dimethylsulfoxide DNJ deoxynojirimicin DOX doxorubicin dt double triplet DTT dithiothreitol EDC 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide EDTA ethylenediaminetetraacetate EEDQ 2-ethoxy-1-ethoxycarbonyl-1,2dihydroquinoline ELISA enzyme-linked immunosorbent assay eq molar equivalents ER endoplasmatic reticulum ERT enzyme replacement therapy ESI electronspray ionization Et ethyl EtOAc ethyl acetate FACS fluorescence-activated cell sorting FCS fetal calf serum FITC fluorescein isothiocyanate iv Fmoc FR FRET FSC FSH(R) FT (e)GFP GBA1 GnRH GPCR GRK GSL h HBTU hCG HCTU HEK293 HMW HOBt HRMS HRP Hsp Hz IGF-II IMDM J LC/MS LH(R) LMW LSD M m M6P M6PC M6Pn m/z mman MC Me (9H-fluoren-9-yl)methoxycarbonyl folate receptor fluorescence resonance energy transfer forward scatter follicle-stimulating hormone (receptor) flow-through (enhanced) green fluorescent protein β-glucocerebrosidase 1 gonadotropin-releasing hormone G protein-coupled receptor G protein-coupled receptor kinase glycosphingolipid hour(s) 2-(1H-Benzotriazole-1-yl)-1,1,3,3tetramethyluronium hexafluorophosphate human chorionic gonadotropin 2-(6-chloro-1H-benzotriazole-1yl)-1,1,3,3-tetramethyluronium hexafluorophosphate human embryonic kidney (cells) high molecular weight N-hydroxybenzotriazole high resolution mass spectrometry horseradish peroxidase heat shock protein hertz insulin-like growth factor-2 Iscove’s modified Dulbecco’s medium coupling constant liquid chromatography/ mass spectrometry luteinizing hormone (receptor) low molecular weight lysosomal storage disorder molar multiplet mannose-6-phosphate mannose-6-phosphate cluster mannose-6-phosphonate mass to charge ratio metamannose mannose cluster methyl MeOH MFI min MPR MR MWCO NBD methanol mean fluorescence intensity minute(s) mannose-6-phosphate receptor mannose receptor molecular weight cutoff 7-nitrobenz-2-oxa-1,3-diazol-4-yl (dye) NBD nucleotide binding domain (protein) NBS newborn bovine calf serum NMP N-methyl-2-pyrrolidone NMR nuclear magnetic resonance NPD Niemann-Pick disease NPY neuropeptide Y NTD N-terminal domain o/n overnight oorthoobsd. observed pparaPAGE polyacrylamide gel electrophoresis PBS phosphate buffered saline PE petroleum ether PEG polyethylene glycol pen/strep penicillin/streptomycin PeT photoinduced electron transfer Ph phenyl ppm parts per million PyBOP (benzotriazol-1-yloxy)tripyrrolidinophosphonium hexafluorophosphate q quartet quant quantitative Rf retention factor rFSH recombinant FSH RhoB rhodamine B dye rpm revolutions per minute rt room temperature s singlet SAR structure-activity relationship sat saturated SBD substrate binding domain (protein) SDS sodium dodecyl sulfate SPPS solid-phase peptide synthesis SrtA sortase A SSC side scatter sst somatostatin receptor t triplet tR retention time TBAF tetrabutylammonium fluoride v TBAI TBDMS TBDPS TBS(T) TEA TEMPO TFA THF THQ TIS tetrabutylammonium iodide tert-butyl dimethylsilyl tert-butyl diphenylsilyl Tris-buffered saline (+ Tween 20) triethylamine 2,2,6,6-tetramethylpiperidinyloxyl trifluoroacetic acid tetrahydrofuran tetrahydroquinoline triisopropyl silane TLC thin layer chromatography Tris 2-amino-2-(hydroxymethyl)-1,3propanediol trz triazole TSH(R) thyroid-stimulating hormone (receptor) WB western blot WT wildtype (e)YFP (enhanced) yellow fluorescent protein Note: The one or three letter codes for the amino acids follow the recommendations of IUPAC. J. Biol. Chem. 1968, 243, 3557-3559 and J. Biol. Chem. 1972, 247, 977-983. vi