2-Amino-3-(phenylsulfanyl)norbornane-2-carboxylate: An Appealing Scaffold for the Design of Rac1–Tiam1 Protein–Protein Interaction Inhibitors

Ruffoni, Alessandro; Ferri, Nicola; Bernini, Sergio Kevin; Ricci, Chiara; Corsini, Alberto; Maffucci, Irene; Clerici, Francesca; Contini, Alessandro

doi:10.1021/jm401924s

Cited by 32 publications

(53 citation statements)

References 40 publications

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“…The intracellular amount of Rac1-GTP and RhoA-GTP were determined by using the G-LISA assay as previously described (Cytoskeleton, Inc Denver, CO, USA) [23][24][25] . 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 61 62 63 64 65 11…”

Section: G-lisa Assay For Rac1 and Rhoamentioning

confidence: 99%

PCSK9 knock-out mice are protected from neointimal formation in response to perivascular carotid collar placement

et al. 2016

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Proprotein convertase subtilisin kexin type 9 (PCSK9) is a keyregulator of hepatic low-density lipoprotein-receptor (LDLR). PCSK9 is expressed in cultured smooth muscle cells (SMCs) and co-localizes with SMCs in human carotid atherosclerotic plaques. The present study aimed at comparing the neointimal lesions induced by periadventitial carotid placement of a non-occlusive collar in PCSK9 knock-out (PCSK9-/-) and wild type (WT) littermate (PCSK9+/+) mice. Collared carotids of the PCSK9-/-mice showed less intimal thickening compared to WT mice (p<0.05), a decreased intimal media ratio (p<0.02) and tendency to higher lumen area. When compared with PCSK9-/-, carotid lesions of WT mice had a higher content of SMCs (p<0.05) and collagen (p<0.05). No difference in macrophage content was detected between the two groups. SMCs freshly isolated from PCSK9-/-, when compared to PCSK9+/+ cells, showed higher levels of the contractile markers α-smooth muscle actin (α-SMA; 2.24±0.36 fold; p<0.01) and myosin heavy chain II (MHC-II; 8.65±1.55 fold; p<0.01), and decreased levels of synthetic markers caldesmon (-54±14%; p<0.01). PCSK9-/-cells also showed in response to platelet-derived growth factor-BB (PDGF-BB) a slower proliferation rate, and impaired migratory capacity and G1/S progression of the cell cycle. The reconstitution of PCSK9 expression, by retroviral infection of PCSK9-/-SMCs, led to a downregulation of α-SMA (-56±2%; p<0.01) and significant induction of caldesmon (1.46±0.3 fold; p<0.05). Proliferation rate of SMCs PCSK9-/-was significantly lower compared to PCSK9 reconstituted cells. Taken together, the present results suggest that by sustaining SMC synthetic phenotype, proliferation and migration PCSK9 may play a pro-atherogenic role in the arterial wall. HighlightsIn response to perivascular manipulation, the neointimal formation is reduced in PCSK9 -/-mice PCSK9 directly regulates smooth muscle cell differentiation, proliferation and migrationThe reconstitution of PCSK9 expression in SMCs PCSK9 -/-determines a switch towards a synthetic phenotype and rescue the impaired proliferation and migration capacities of SMCs PCSK9 -/-SMCs PCSK9 -/-respond less efficiently to the proliferative and chemotactic action of PDGF-BB 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 61 62 63 64 65 2 AbstractProprotein convertase subtilisin kexin type 9 (PCSK9) is a key-regulator of hepatic low-density lipoprotein-receptor (LDLR). PCSK9 is expressed in cultured smooth muscle cells (SMCs) and colocalizes with SMCs in human carotid atherosclerotic plaques. The present study aimed at comparing the neointimal lesions induced by periadventitial carotid placement of a non-occlusive collar in PCSK9 knock-out (PCSK9 -/-) and wild type (WT) littermate (PCSK9 +/+ ) mice. Collared carotids of the PCSK9 -/-mice showed less intimal thickening compared...

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Section: G-lisa Assay For Rac1 and Rhoamentioning

confidence: 99%

PCSK9 knock-out mice are protected from neointimal formation in response to perivascular carotid collar placement

et al. 2016

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“…For the experiments cells were seeded at a density of 2 ×10 5 /35 mm Petri dish and incubated with DMEM supplemented with 10% FCS; 24 hours later the medium was changed to one containing 0.4% FCS, and the cultures were incubated for 48 hours. At this time, the peptides were added to the cultured medium, and after 4 hours the intracellular amounts of Rac1‐GTP were determined by using the G‐LISA assay, as previously described…”

Section: Methodsmentioning

confidence: 99%

“…This small molecule fits into the surface groove of Rac1 involved in the binding with GEFs, thus interfering with the Tiam1‐Rac1 interaction in a selective manner, without interfering with other small G proteins, such as Cdc‐42 and RhoA. From this pioneer observation, additional and more potent selective Rac1 inhibitors have been identified . A second approach to selectively interfere with the cellular function of small G proteins is represented by the identification of compounds capable to interfere with the PPI between the small G protein and their effectors.…”

Section: Introductionmentioning

confidence: 99%

Peptide modulators of Rac1/Tiam1 protein‐protein interaction: An alternative approach for cardiovascular diseases

et al. 2018

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Rac1 GTPase interaction with guanine nucleotide exchange factor Tiam1 is involved in several cancer types and cardiovascular diseases. Although small molecules interfering with their protein-protein interaction (PPI) were identified and studied, the ability of small peptides and peptide mimics acting as Rac1/Tiam1 PPI inhibitors has not been yet explored. Using computational alanine scanning (CAS), the "hot" interfacial residues have been determined allowing the design of a small library of putative PPI inhibitors. In particular, the insertion of an unnatural alpha, alpha disubstituted amino acid, that is norbornane amino acid, and the side chain stapling have been evaluated regarding both conformational stability and biological activity. REMD calculations and CD studies have indicated that one single norbornane amino acid at the N-terminus is not sufficient to stabilize the helix structure, while the side-chain stapling is a more efficient strategy. Furthermore, both engineered peptides have been found able to reduce Rac1-GTP levels in cultured human smooth muscle cells, while wild type sequence is not active.

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“…41 had an IC 50 value of 4 μ m for the antiproliferation of F3II cancer cells, was more potent in blocking GEF activation in vitro than 40 , and caused a 60 % reduction in metastatic lung colonies in vivo. A different approach used de novo design to yield an inhibitor that would have the same pharmacophoric features as 38 and other previous compounds but a unrelated scaffold . In cellular assays, 42 reduced Rac1‐GTP levels with an IC 50 value of 2.5 μ m albeit containing a dianiline structural alert motif.…”

Section: Rho Gtpasesmentioning

confidence: 99%

Targeting the Small GTPase Superfamily through Their Regulatory Proteins

2020

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The Ras superfamily of small GTPases are guanine‐nucleotide‐dependent switches essential for numerous cellular processes. Mutations or dysregulation of these proteins are associated with many diseases, but unsuccessful attempts to target the small GTPases directly have resulted in them being classed as “undruggable”. The GTP‐dependent signaling of these proteins is controlled by their regulators; guanine nucleotide exchange factors (GEFs), GTPase activating proteins (GAPs), and in the Rho and Rab subfamilies, guanine nucleotide dissociation inhibitors (GDIs). This review covers the recent small molecule and biologics strategies to target the small GTPases through their regulators. It seeks to critically re‐evaluate recent chemical biology practice, such as the presence of PAINs motifs and the cell‐based readout using compounds that are weakly potent or of unknown specificity. It highlights the vast scope of potential approaches for targeting the small GTPases in the future through their regulatory proteins.

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2-Amino-3-(phenylsulfanyl)norbornane-2-carboxylate: An Appealing Scaffold for the Design of Rac1–Tiam1 Protein–Protein Interaction Inhibitors

Cited by 32 publications

References 40 publications

PCSK9 knock-out mice are protected from neointimal formation in response to perivascular carotid collar placement

PCSK9 knock-out mice are protected from neointimal formation in response to perivascular carotid collar placement

Peptide modulators of Rac1/Tiam1 protein‐protein interaction: An alternative approach for cardiovascular diseases

Targeting the Small GTPase Superfamily through Their Regulatory Proteins

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