Molecular Clip and Tweezer Introduce New Mechanisms of Enzyme Inhibition

Talbiersky, Peter; Bastkowski, Frank; Klärner, Frank‐Gerrit; Schräder, Thomas

doi:10.1021/ja801441j

Cited by 140 publications

(171 citation statements)

References 26 publications

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“…However, this was found not Lys residues with micromolar affinity, which is relatively weak but has been found to be sufficient for modulating the assembly and inhibit the toxicity of multiple amyloidogenic proteins without causing toxicity in cell culture unless substantially higher concentrations were used [69]. These data are consistent with in vitro enzyme inhibition studies, which required MT concentrations at least an order of magnitude higher than those needed for inhibition of amyloidogenic proteins [68,70] and with data showing beneficial effects with no associated toxicity in animal models [71,72].…”

supporting

confidence: 71%

“…In blood-brain-barrier studies, one hour following intravenous injection, brain levels of 3 H-CLR01, measured by scintillation counting, were ~2% of the levels found in the blood [71]. Importantly, the binding of CLR01 is highly labile and the affinity of the compound for Lys is in the micromolar range [68,69], although higher affinity may be achieved in certain cases, depending on the number of Lys residues in the target protein and the specific sequence of the protein [134]. The high lability and moderate affinity prevent …”

Section: Effects On Ttr Toxicity In Vivomentioning

confidence: 98%

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Modulators of amyloid protein aggregation and toxicity: EGCG and CLR01

Attar

Rahimi

Bitan

2013

Translational Neuroscience

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Abnormal protein folding and self-assembly causes over 30 cureless human diseases for which no diseasemodifying therapies are available. The common side to all these diseases is formation of aberrant toxic protein oligomers and amyloid fibrils. Both types of assemblies are drug targets, yet each presents major challenges to drug design, discovery, and development. In this review, we focus on two small molecules that inhibit formation of toxic amyloid protein assemblies -the green-tea derivative (-)-epigallocatechin-3-gallate (EGCG), which was identified through a combination of epidemiologic data and a compound library screen, and the molecular tweezer CLR01, whose inhibitory activity was discovered in our group based on rational reasoning, and subsequently confirmed experimentally. Both compounds act in a manner that is not specific to one particular protein and thus are useful against a multitude of amyloidogenic proteins, yet they act via distinct putative mechanisms. CLR01 disrupts protein aggregation through specific binding to lysine residues, whereas the mechanisms underlying the activity of EGCG are only recently beginning to unveil. We discuss current in vitro and, where available, in vivo literature related to EGCG and CLR01's effects on amyloid β-protein, α-synuclein, transthyretin, islet amyloid polypeptide, and calcitonin. We also describe the toxicity, pharmacokinetics, and mechanism of action of each compound.

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supporting

confidence: 71%

Section: Effects On Ttr Toxicity In Vivomentioning

confidence: 98%

Modulators of amyloid protein aggregation and toxicity: EGCG and CLR01

Attar

Rahimi

Bitan

2013

Translational Neuroscience

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“…Fluorescence titration yielded K d = 8.69 μM (Supplementary Figure S6), similar to values reported previously for N-terminal K residues. 46 In contrast, no change was observed in the spectrum of IAPP 2−7 even in the presence of 3-fold excess CLR01 (Supplementary Figure S7), indicating that the presence of K1 was essential for CLR01 binding to IAPP 1−7 .…”

Section: Acs Chemical Biologymentioning

confidence: 95%

Molecular Tweezers Inhibit Islet Amyloid Polypeptide Assembly and Toxicity by a New Mechanism

Lopes¹,

Attar²,

Nair³

et al. 2015

ACS Chem. Biol.

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In type-2 diabetes (T2D), islet amyloid polypeptide (IAPP) self-associates into toxic assemblies causing islet β-cell death. Therefore, preventing IAPP toxicity is a promising therapeutic strategy for T2D. The molecular tweezer CLR01 is a supramolecular tool for selective complexation of K residues in (poly)peptides. Surprisingly, it inhibits IAPP aggregation at substoichiometric concentrations even though IAPP has only one K residue at position 1, whereas efficient inhibition of IAPP toxicity requires excess CLR01. The basis for this peculiar behavior is not clear. Here, a combination of biochemical, biophysical, spectroscopic, and computational methods reveals a detailed mechanistic picture of the unique dual inhibition mechanism for CLR01. At low concentrations, CLR01 binds to K1, presumably nucleating nonamyloidogenic, yet toxic, structures, whereas excess CLR01 binds also to R11, leading to nontoxic structures. Encouragingly, the CLR01 concentrations needed for inhibition of IAPP toxicity are safe in vivo, supporting its development toward diseasemodifying therapy for T2D.

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“…The Na + salts of CLR01 and CLR03 were prepared and purified analogously to the Li salts, as described previously (16).…”

Section: Methodsmentioning

confidence: 99%

The Lys-Specific Molecular Tweezer, CLR01, Modulates Aggregation of the Mutant p53 DNA Binding Domain and Inhibits Its Toxicity

et al. 2015

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The tumor suppressor p53 plays a unique role as a central hub of numerous cell proliferation and apoptotic pathways and its malfunction due to mutations is a major cause of various malignancies. Therefore, it serves as an attractive target for developing novel anti-cancer therapeutics. Due to its intrinsically unstable DNA-binding domain, p53 unfolds rapidly at physiological temperature. Certain mutants shift the equilibrium towards the unfolded state and yield high-molecular-weight, non-functional, and cytotoxic β-sheet-rich aggregates that share tinctorial and conformational similarities with amyloid deposits found in various protein-misfolding diseases. Here, we examined the effect of a novel protein-assembly modulator, the lysine (Lys)-specific molecular tweezer, CLR01, on different aggregation stages of misfolded mutant p53 in vitro and on the cytotoxicity of the resulting p53 aggregates in cell culture. We found that CLR01 induced rapid formation of β-sheet-rich, intermediate-size p53 aggregates, yet inhibited further p53 aggregation and reduced cytotoxicity of the resulting aggregates. Our data suggest that aggregation modulators, such as CLR01, could prevent formation of toxic p53 aggregates.

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Molecular Clip and Tweezer Introduce New Mechanisms of Enzyme Inhibition

Cited by 140 publications

References 26 publications

Modulators of amyloid protein aggregation and toxicity: EGCG and CLR01

Modulators of amyloid protein aggregation and toxicity: EGCG and CLR01

Molecular Tweezers Inhibit Islet Amyloid Polypeptide Assembly and Toxicity by a New Mechanism

The Lys-Specific Molecular Tweezer, CLR01, Modulates Aggregation of the Mutant p53 DNA Binding Domain and Inhibits Its Toxicity

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