Ultrasensitive small molecule fluorogenic probe for human heparanase

Li, Jun; Schleyer, Kelton A.; Bryan, Tyrel; Xie, Changjian; Seabra, Gustavo; Xu, Yongmei; Kafle, Arjun; Cui, Chao; Wang, Ying; Yin, Kunlun; Fetrow, Benjamin; Henderson, Paul K. P.; Fatland, Peter Zannes; Liu, Jian; Li, Chenglong; Guo, Hua; Cui, Lina

doi:10.1039/d0sc04872k

Cited by 14 publications

(28 citation statements)

References 54 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The structure revealed previously unknown interactions involved in enzymatic processing and suggested using substrates with better leaving groups, e. g., halogenated coumarins. Subsequently, Cui and coworkers [73] reported that difluorocoumarins such as 36 are more efficient aglycone leaving groups for heparanase substrates, in line with our hypotheses.…”

Section: Tools For Studying Hs‐protein Interactionssupporting

confidence: 88%

From Cancer to COVID‐19: A Perspective on Targeting Heparan Sulfate‐Protein Interactions

Chhabra

Doherty

See

et al. 2021

The Chemical Record

View full text Add to dashboard Cite

Heparan sulfate (HS) is a complex, polyanionic polysaccharide ubiquitously expressed on cell surfaces and in the extracellular matrix. HS interacts with numerous proteins to mediate a vast array of biological and pathological processes. Inhibition of HS‐protein interactions is thus an attractive approach for new therapeutic development for cancer and infectious diseases, including COVID‐19; however, synthesis of well‐defined native HS oligosaccharides remains challenging. This has aroused significant interest in the development of HS mimetics which are more synthetically tractable and have fewer side effects, such as undesired anticoagulant activity. This account provides a perspective on the design and synthesis of different classes of HS mimetics with useful properties, and the development of various assays and molecular modelling tools to progress our understanding of their interactions with HS‐binding proteins.

show abstract

Section: Tools For Studying Hs‐protein Interactionssupporting

confidence: 88%

From Cancer to COVID‐19: A Perspective on Targeting Heparan Sulfate‐Protein Interactions

Chhabra

Doherty

See

et al. 2021

The Chemical Record

View full text Add to dashboard Cite

show abstract

“…Recently, we reported the first structurally defined ultrasensitive fluorogenic probe HADP (1) for detecting heparanase activity with high selectivity and sensitivity, which we used in a high-throughput screen for novel heparanase inhibitors. 15 In our initial probe design, use of the fluorogenic reporter 4-methylumbelliferone (4-MU) did not facilitate turnover of the probe by HPSE, consistent with the exclusively endo -glycosidic selectivity of HPSE enzymatic activity. However, by incorporating two electron-withdrawing fluorine atoms on the methylumbelliferone reporter, ortho to the phenolic oxygen, our HADP probe successfully elicited exo -glycosidic activity from HPSE.…”

Section: Maintextmentioning

confidence: 72%

“…However, by incorporating two electron-withdrawing fluorine atoms on the methylumbelliferone reporter, ortho to the phenolic oxygen, our HADP probe successfully elicited exo -glycosidic activity from HPSE. 15 Following this study, we developed a second fluorogenic probe for imaging heparanase activity in living cells (J.L., Z.W., and L.C., unpublished results), which also required two ortho -position fluorine atoms installed on the reporter, to elicit HPSE activity. Encouraged by these advances, we attempted to develop a near-infrared (NIR) probe for monitoring heparanase in vivo by extending the conjugation system of fluorophore, but the synthesis failed due to incompatibility of the NIR fluorophore with the chemical manipulation of the disaccharide recognition unit.…”

Section: Maintextmentioning

confidence: 99%

“…This was due to the resultant weakening of the glycosidic bond and lowering of the transition state energy for enzymatic turnover by heparanase. 15 Based on these results, we deigned two molecules bearing a 4-hydroxybenzyl alcohol linker substituted with either two or four fluorine atoms to facilitate enzymatic cleavage by heparanase. A simple coumarin as fluorescent reporter to examine whether the linker can be disassembled upon incubation with heparanase.…”

Section: Maintextmentioning

confidence: 99%

See 1 more Smart Citation

A self-immolative linker for heparanase activatable probes

Schleyer

Liu

Wang

et al. 2021

Preprint

Self Cite

View full text Add to dashboard Cite

Substrate-based probes utilize known substrate specificity parameters to create a probe that can be activated by a target enzyme. In developing probes for heparanase, an endo-beta-glucuronidase, we previously reported that small, inactive substrate-based probes could be electronically tuned by incorporating electron-withdrawing atoms on the aromatic aglycone fluorophore, ortho- to the cleaved glycosidic bond. However, the installation of electron-withdrawing groups directly onto established fluorophores or other reporters complicates the synthesis of new heparanase probes. In this work we report a new design strategy to expand the toolkit of heparanase imaging probes, in which the installation of an electronically tuned benzyl alcohol linker restored the activity of a previously inactive heparanase probe using 4-methylumbelliferone as the fluorescent reporter, suggesting such a linker can provide a scaffold for facile development of activatable heparanase probes bearing a variety of imaging moieties.

show abstract

“…A 7-hydroxymethylcoumarin moiety was selected as the donor fluorophore, due to the possibility of functionalization onto a porphyrin scaffold and the excellent spectral overlap with the absorbance spectrum of porphyrin-based compounds (Figures S1 and S2). Additionally, 7-hydroxymethylcoumarin derivatives are also known to be fairly photostable, , including compound 1 (Figure S3), a characteristic that will importantly maximize the observed increase in fluorescence following enzymatic probe cleavage.…”

Section: Introductionmentioning

confidence: 99%

A Coumarin–Porphyrin FRET Break-Apart Probe for Heme Oxygenase-1

et al. 2021

View full text Add to dashboard Cite

Heme oxygenase-1 (HO-1) is a vital enzyme in humans that primarily regulates free heme concentrations. The overexpression of HO-1 is commonly associated with cardiovascular and neurodegenerative diseases including atherosclerosis and ischemic stroke. Currently, there are no known chemical probes to detect HO-1 activity, limiting its potential as an early diagnostic/prognostic marker in these serious diseases. Reported here are the design, synthesis, and photophysical and biological characterization of a coumarin–porphyrin FRET break-apart probe to detect HO-1 activity, Fe–L 1 . We designed Fe–L 1 to “break-apart” upon HO-1-catalyzed porphyrin degradation, perturbing the efficient FRET mechanism from a coumarin donor to a porphyrin acceptor fluorophore. Analysis of HO-1 activity using Escherichia coli lysates overexpressing hHO-1 found that a 6-fold increase in emission intensity at 383 nm was observed following incubation with NADPH. The identities of the degradation products following catabolism were confirmed by MALDI-MS and LC–MS, showing that porphyrin catabolism was regioselective at the α-position. Finally, through the analysis of Fe–L 2 , we have shown that close structural analogues of heme are required to maintain HO-1 activity. It is anticipated that this work will act as a foundation to design and develop new probes for HO-1 activity in the future, moving toward applications of live fluorescent imaging.

show abstract

Ultrasensitive small molecule fluorogenic probe for human heparanase

Abstract: Heparanase (HPA) is a critical enzyme involved in the remodeling of the extracellular matrix (ECM), and its elevated expression has been linked with diseases such as various types of cancer...

Cited by 14 publications

References 54 publications

From Cancer to COVID‐19: A Perspective on Targeting Heparan Sulfate‐Protein Interactions

From Cancer to COVID‐19: A Perspective on Targeting Heparan Sulfate‐Protein Interactions

A self-immolative linker for heparanase activatable probes

A Coumarin–Porphyrin FRET Break-Apart Probe for Heme Oxygenase-1

Contact Info

Product

Resources

About