Potent Inhibitors of β-Tryptase and Human Leukocyte Elastase Based on the MCoTI-II Scaffold

Thongyoo, Panumart; Bonomelli, Camille; Leatherbarrow, Robin J.; Tate, Edward W.

doi:10.1021/jm901233u

Cited by 126 publications

(132 citation statements)

References 26 publications

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“…This observation is consistent with recent studies which demonstrated that grafting sequences as long as sixteen amino acid residues into loop 6 does not alter the disulfide bond connectivity within MCoTI [29]. Moreover, the MCoTI-ll scaffold has been engineered to bind to a range of protein targets including HMD2 [29], beta-tryptase, human leukocyte elastase and matriptase [16,27,30,31], suggesting that the framework itself has plasticity to accommodate different sequences. However, introducing the Sortase A recognition sequence into other loop regions within the framework might pose a problem and hence would need to be carefully assessed on a case-by-case basis.…”

Section: Resultssupporting

confidence: 89%

Backbone cyclization of a recombinant cystine‐knot peptide by engineered Sortase A

Stanger¹,

Maurer²,

Kaluarachchi³

et al. 2014

FEBS Letters

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Edited by Miguel De la RosaKeywords: Cyclotide Recombinant expression Sortase Protein engineering MCoTI-II Cystine-knot peptide a b s t r a c t Cyclotides belong to the family of cyclic cystine-knot peptides and have shown promise as scaffolds for protein engineering and pharmacological modulation of cellular protein activity. Cyclotides are characterized by a cystine-knotted topology and a head-to-tail cyclic polypeptide backbone. While they are primarily produced in plants, cyclotides have also been obtained by chemical synthesis. However, there is still a need for methods to generate cyclotides in high yields to near homogeneity. Here, we report a biomimetic approach which utilizes an engineered version of the enzyme Sortase A to catalyze amide backbone cyclization of the recombinant cyclotide MCoTI-II, thereby allowing the efficient production of active homogenous species in high yields. Our results provide proof of concept for using engineered Sortase A to produce cyclic MCoTI-II and should be generally applicable to generating other cyclic cystine-knot peptides.

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Section: Resultssupporting

confidence: 89%

Backbone cyclization of a recombinant cystine‐knot peptide by engineered Sortase A

Stanger¹,

Maurer²,

Kaluarachchi³

et al. 2014

FEBS Letters

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“…The elastase inhibitory potency of synthetic compounds was determined by Thongyoo et al, 2009 method with some changes. The quantity of free p-nitroaniline, that was hydrolyzed by the action of elastase from the substrate (N-succinyl-Ala-Ala-Ala-p-nitroanilide) was quantified by calculating the absorbance at 410 nm.…”

Section: Elastase Inhibition Activity Assaymentioning

confidence: 99%

Synthesis of 4-aryl-2,6-dimethyl-3,5-bis-N-(aryl)-carbamoyl-1,4-dihydropyridines as novel skin protecting and anti-aging agents

Saeed

Shahzad

Larik

et al. 2017

Bangladesh J Pharmacol

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A series of 4-aryl-2,6-dimethyl-3,5-bis-N-(aryl)-carbamoyl-1,4-dihydropyri-dines 6a-6h were prepared by using the one-pot three component synthetic method. The target compounds 6a-6h were synthesized by reacting two molar equivalents of ketone functionality and one mole of aromatic aldehydes in ammonium acetate to obtain the desired products. The structures of newly synthesized compounds were characterized by FT-IR, 1H-NMR, 13C-NMR, and elemental analysis. All the synthesized compounds were screened for their elastase inhibition and antioxidant activity. Almost all of the com-pounds 6a-h showed good to excellent activities against elastase enzyme more than the reference drug. Compounds 6d and 6b at 0.2 ± 0.0 µM and 0.2 ± 0.0 µM were found to most potent derivatives against elastase enzyme. Compound 6a exhibited prominent free radical scavenging activity. From the results of the biological activity, we infer that some derivatives can serve as lead molecules in pharmacology.Video Clip of Methodology:3 min 13 sec <a href="https://youtube.com/v/gPLdpGpZhR8">Full Screen</a> <a href="https://youtube.com/watch?v=gPLdpGpZhR8">Alternate</a>

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“…In a more recent study, the same authors also generated inhibitors of the serine proteases β-tryptase and human leukocyte elastase (HLE) using the backbone of MCoTI-II [96]. β-Tryptase is implicated in allergic and inflammatory disorders, and HLE has been associated with respiratory and pulmonary disorders.…”

Section: Designing Cyclotides With Novel Biological Activitiesmentioning

confidence: 99%

“…β-Tryptase is implicated in allergic and inflammatory disorders, and HLE has been associated with respiratory and pulmonary disorders. Replacing the P1 residue in loop 1 produced several MCoTI-II mutants (K6A and K6V) with activity against HLE with K i values of 20-30 nM [96] and K i values against trypsin above 1 μM. Removal of the SDGG peptide segment in loop 6 yielded a ß-tryptase inhibitor with a K i ≈ 10 nM without significantly altering the three-dimensional structure as determined by NMR [96].…”

Section: Designing Cyclotides With Novel Biological Activitiesmentioning

confidence: 99%

“…Replacing the P1 residue in loop 1 produced several MCoTI-II mutants (K6A and K6V) with activity against HLE with K i values of 20-30 nM [96] and K i values against trypsin above 1 μM. Removal of the SDGG peptide segment in loop 6 yielded a ß-tryptase inhibitor with a K i ≈ 10 nM without significantly altering the three-dimensional structure as determined by NMR [96]. The authors hypothesized that deletion of the aspartic acid residue in MCoTI-II should improve activity by removal of repulsive electrostatic interactions with β-tryptase thus improving the inhibitory constant against trypsin 160-fold when compared to the wild-type MCoTI-II.…”

Section: Designing Cyclotides With Novel Biological Activitiesmentioning

confidence: 99%

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Biological Activities of Natural and Engineered Cyclotides, a Novel Molecular Scaffold for Peptide-Based Therapeutics

Garcia¹,

Camarero²

2010

CMP

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Cyclotides are a growing family of large plant-derived backbone-cyclized polypeptides (≈30 amino acids long) that share a disulfide-stabilized core characterized by an unusual knotted structure. Their unique circular backbone topology and knotted arrangement of three disulfide bonds makes them exceptionally stable to thermal, chemical, and enzymatic degradation compared to other peptides of similar size. Currently more than 100 sequences of different cyclotides have been characterized and the number is expected to increase dramatically in the coming years. Considering their stability, biological activities and ability to cross the cell membrane, cyclotides can be exploited to develop new peptide-based drugs with high potential for success. The cyclotide scaffold can be engineered or evolved using molecular evolution to inhibit proteinprotein interactions implicated in cancer and other human diseases, or design new antimicrobial. The present review reports the biological diversity and therapeutic potential of natural and engineered cyclotides.

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Potent Inhibitors of β-Tryptase and Human Leukocyte Elastase Based on the MCoTI-II Scaffold

Cited by 126 publications

References 26 publications

Backbone cyclization of a recombinant cystine‐knot peptide by engineered Sortase A

Backbone cyclization of a recombinant cystine‐knot peptide by engineered Sortase A

Synthesis of 4-aryl-2,6-dimethyl-3,5-bis-N-(aryl)-carbamoyl-1,4-dihydropyridines as novel skin protecting and anti-aging agents

Biological Activities of Natural and Engineered Cyclotides, a Novel Molecular Scaffold for Peptide-Based Therapeutics

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