A Novel “Molecular Tweezer” Inhibitor of α-Synuclein Neurotoxicity in Vitro and in Vivo

Prabhudesai, Shubhangi; Sinha, Sharmistha; Attar, Aida; Kotagiri, Aswani; Fitzmaurice, Arthur G.; Lakshmanan, Rajeswari; Ivanova, Magdalena I.; Loo, Joseph A.; Klärner, Frank‐Gerrit; Schräder, Thomas; Stahl, Mark; Bitan, Gal; Bronstein, Jeff M.

doi:10.1007/s13311-012-0105-1

Cited by 152 publications

(267 citation statements)

References 55 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: 66%

“…Addition of CLR01 to the water environment of the fish at 1 or 10 μM caused dramatic, dose-dependent improvements in both the body phenotype and the survival of the fish [72]. Immunohistochemical analysis of the zebra fish neurons showed abundant aggregated α-synuclein in untreated fish, whereas only soluble α-synuclein was observed in fish treated with CLR01.…”

Section: Clr01mentioning

confidence: 99%

“…The effect of CLR01 on α-synuclein aggregation was investigated using ThT fluorescence, EM, and native PAGE [72].…”

Section: Clr01mentioning

confidence: 99%

“…CLR01: CLR01's in vivo effects on α-synuclein aggregation and toxicity were first tested in a zebra fish (Brachydanio rerio) model, which expresses human wild-type α-synuclein under a neuronal promoter that causes a deformed body phenotype due to neuronal apoptosis, and early death [72]. Addition of CLR01 to the water environment of the fish at 1 or 10 μM caused dramatic, dose-dependent improvements in both the body phenotype and the survival of the fish [72].…”

Section: Clr01mentioning

confidence: 99%

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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: 66%

Section: Clr01mentioning

confidence: 99%

“…The effect of CLR01 on α-synuclein aggregation was investigated using ThT fluorescence, EM, and native PAGE [72].…”

Section: Clr01mentioning

confidence: 99%

Section: Clr01mentioning

confidence: 99%

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Modulators of amyloid protein aggregation and toxicity: EGCG and CLR01

Attar

Rahimi

Bitan

2013

Translational Neuroscience

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“…Of the amyloidogenic proteins for which CLR01's inhibitory activity was studied previously (14,27,36,37), Aβ40, Aβ42, tau, α-synuclein and islet amyloid polypeptide are intrinsically disordered, insulin, β 2 -microglobulin, and TTR are natively structured, and calcitonin is a relatively short peptide that is partially 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 relative to the structured proteins, as would be expected for a partially structured peptide (14).…”

Section: Clr01 Reduces the Toxicity Of P53dbd Aggregatesmentioning

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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Intrinsically Disordered Proteins

Uversky

2020

Structural Biology in Drug Discovery

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Many functional proteins do not have well defined folded structures. In recent years, both experimental and computational approaches have been developed to study the conformational behaviour of this type of protein. It has been shown previously that experimental RDCs (residual dipolar couplings) can be used to study the backbone sampling of disordered proteins in some detail. In these studies, the backbone structure was modelled using a common geometry for all amino acids. In the present paper, we demonstrate that experimental RDCs are also sensitive to the specific geometry of each amino acid as defined by energyminimized internal coordinates. We have modified the FM (flexible-Meccano) algorithm that constructs conformational ensembles on the basis of a statistical coil model, to account for these differences. The modified algorithm inherits the advantages of the FM algorithm to efficiently sample the potential energy landscape for coil conformations. The specific geometries incorporated in the new algorithm result in a better reproduction of experimental RDCs and are generally applicable for further studies to characterize the conformational properties of intrinsically disordered proteins. In addition, the internal-coordinate based algorithm is an order of magnitude more efficient, and facilitates side-chain construction, surface osmolyte simulation, spin-label distribution sampling and proline cis/trans isomer simulation.

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A Novel “Molecular Tweezer” Inhibitor of α-Synuclein Neurotoxicity in Vitro and in Vivo

Cited by 152 publications

References 55 publications

Modulators of amyloid protein aggregation and toxicity: EGCG and CLR01

Modulators of amyloid protein aggregation and toxicity: EGCG and CLR01

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

Intrinsically Disordered Proteins

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