ABSTRACT
Vascular-disrupting agents (VDA) specifically target established neovasculature which results in vascular shutdown. This therapeutic strategy could improve the outcome of pathologies involving aberrant angiogenesis. Although several classes of VDA exist, inhibitors of tubulin assembly (ITA) represent the main category. A series of 21 conformationnally-restricted analogues of E7010, a known ITA-VDA, were designed and synthesised as novel inhibitors of tubulin assembly (ITA) and vascular-disrupting agents (VDA). Among them, indole 4j exhibited good potency against HUVEC and HIG-82 cell lines, as well as a good ability to inhibit tubulin assembly. Furthermore, indole 4j reduced HUVEC migration in a dose-dependent manner, indicating a vascular disrupting activity comparable to that of the gold standard, Combretastatin A4 (CA4).
Subject(s)
Antineoplastic Agents , Tubulin , Tubulin/metabolism , Cell Line, Tumor , Tubulin Modulators , Antineoplastic Agents/pharmacology , Indoles/pharmacology , Angiogenesis Inhibitors/pharmacologyABSTRACT
BACKGROUND AND PURPOSE: Opioids are effective painkillers. However, their risk-benefit ratio is dampened by numerous adverse effects and opioid misuse has led to a public health crisis. Safer alternatives are required, but isolating the antinociceptive effect of opioids from their adverse effects is a pharmacological challenge because activation of the µ opioid receptor triggers both the antinociceptive and adverse effects of opioids. EXPERIMENTAL APPROACH: The TREK1 potassium channel is activated downstream of µ receptor and involved in the antinociceptive activity of morphine but not in its adverse effects. Bypassing the µ opioid receptor to directly activate TREK1 could therefore be a safer analgesic strategy. KEY RESULTS: We developed a selective TREK1 activator, RNE28, with antinociceptive activity in naive rodents and in models of inflammatory and neuropathic pain. This activity was lost in TREK1 knockout mice or wild-type mice treated with the TREK1 blocker spadin, showing that TREK1 is required for the antinociceptive activity of RNE28. RNE28 did not induce respiratory depression, constipation, rewarding effects, or sedation at the analgesic doses tested. CONCLUSION AND IMPLICATIONS: This proof-of-concept study shows that TREK1 activators could constitute a novel class of painkillers, inspired by the mechanism of action of opioids but devoid of their adverse effects.
Subject(s)
Analgesics, Opioid , Neuralgia , Analgesics , Analgesics, Opioid/adverse effects , Animals , Mice , Mice, Knockout , Morphine , Receptors, Opioid, muABSTRACT
A short and efficient synthesis, based on a one-step double elimination, of a key intermediate in the synthesis of various glucocorticosteroids has been developed. This method can be carried out on large scale for further industrial applications. The synthesis allowed us to identify a novel prednisolone derivative 10 and its anti-inflammatory activity was determined in an in vivo model of inflammation. In order to understand the regioselectivity of the double elimination under various conditions, mechanistic studies were undertaken and confirmed the experimental results. We also propose a mechanism for the formation of the new steroid 10 studied by molecular modeling.
Subject(s)
Glucocorticoids/chemistry , Glucocorticoids/chemical synthesis , Animals , Anti-Inflammatory Agents/chemical synthesis , Anti-Inflammatory Agents/chemistry , Anti-Inflammatory Agents/pharmacology , Anti-Inflammatory Agents/therapeutic use , Chemistry Techniques, Synthetic , Edema/drug therapy , Glucocorticoids/pharmacology , Glucocorticoids/therapeutic use , Male , Mice , Models, Molecular , Molecular Conformation , Prednisolone/chemical synthesis , Prednisolone/chemistry , Prednisolone/pharmacology , Prednisolone/therapeutic useABSTRACT
The TWIK-related K+ channel, TREK-1, has recently emerged as an attractive therapeutic target for the development of a novel class of analgesic drugs, suggesting that activation of TREK-1 could result in pain inhibition. Here, we report the synthesis of a series of substituted acrylic acids (1-54) based on our previous work with caffeate esters. The analogues were evaluated for their ability to modulate TREK-1 channel by electrophysiology and for their in vivo antinociceptive activity (acetic acid-induced writhing and hot plate assays), leading to the identification of a series of novel molecules able to activate TREK-1 and displaying potent antinociceptive activity in vivo. Furyl analogue 36 is the most promising of the series.
Subject(s)
Analgesics/pharmacology , Potassium Channels, Tandem Pore Domain/agonists , AnimalsABSTRACT
Potassium (K(+)) channels are membrane proteins expressed in most living cells that selectively control the flow of K(+) ions. More than 80 genes encode the K(+) channel subunits in the human genome. The TWIK-related K(+) channel (TREK-1) belongs to the two-pore domain K(+) channels (K2P) and displays various properties including sensitivity to physical (membrane stretch, acidosis, temperature) and chemical stimuli (signaling lipids, volatile anesthetics). The distribution of TREK-1 in the central nervous system, coupled with the physiological consequences of its opening and closing, leads to the emergence of this channel as an attractive therapeutic target. We review the TREK-1 channel, its structural and functional properties, and the pharmacological agents (agonists and antagonists) able to modulate its gating.
Subject(s)
Neuroprotective Agents/pharmacology , Potassium Channels, Tandem Pore Domain/agonists , Potassium Channels, Tandem Pore Domain/antagonists & inhibitors , Arrhythmias, Cardiac/drug therapy , Depression/drug therapy , Epilepsy/drug therapy , Humans , Inflammation/drug therapy , Models, Molecular , Molecular Structure , Neuroprotective Agents/chemistry , Pain/drug therapy , Potassium Channels, Tandem Pore Domain/metabolism , Structure-Activity RelationshipABSTRACT
ZL006 and IC87201 have been presented as efficient inhibitors of the nNOS/PSD-95 protein-protein interaction and shown great promise in cellular experiments and animal models of ischemic stroke and pain. Here, we investigate the proposed mechanism of action of ZL006 and IC87201 using biochemical and biophysical methods, such as fluorescence polarization (FP), isothermal titration calorimetry (ITC), and (1)H-(15)N HSQC NMR. Our data show that under the applied in vitro conditions, ZL006 and IC87201 do not interact with the PDZ domains of nNOS or PSD-95, nor inhibit the nNOS-PDZ/PSD-95-PDZ interface by interacting with the ß-finger of nNOS-PDZ. Our findings have implications for further medicinal chemistry efforts of ZL006, IC87201 and analogues, and challenge the general and widespread view on their mechanism of action.
Subject(s)
Aminosalicylic Acids/pharmacology , Benzylamines/pharmacology , Chlorophenols/pharmacology , Intracellular Signaling Peptides and Proteins/antagonists & inhibitors , Membrane Proteins/antagonists & inhibitors , Nitric Oxide Synthase Type I/antagonists & inhibitors , PDZ Domains/drug effects , Triazoles/pharmacology , Calorimetry , Disks Large Homolog 4 Protein , Fluorescence Polarization , Humans , Magnetic Resonance SpectroscopyABSTRACT
The TWIK-related K(+) channel, TREK-1, has recently emerged as an attractive therapeutic target for the development of a novel class of analgesic drugs. It has been reported that TREK-1 -/- mice were more sensitive than wild-type mice to painful stimuli, suggesting that activation of TREK-1 could result in pain inhibition. Here we report the synthesis of a series of substituted caffeate esters (12a-u) based on the hit compound CDC 2 (cinnamyl 3,4-dihydroxyl-α-cyanocinnamate). These analogs were evaluated for their ability to modulate TREK-1 channel by electrophysiology and for their in vivo antinociceptive activity (acetic acid induced-writhing assay) leading to the identification a series of novel molecules able to activate TREK-1 and displaying potent analgesic activity in vivo.
Subject(s)
Analgesics/chemistry , Analgesics/therapeutic use , Caffeic Acids/chemistry , Caffeic Acids/therapeutic use , Pain/drug therapy , Potassium Channels, Tandem Pore Domain/metabolism , Analgesics/pharmacology , Animals , Caffeic Acids/pharmacology , Cinnamates/chemistry , Cinnamates/pharmacology , Cinnamates/therapeutic use , Esters/chemistry , Esters/pharmacology , Esters/therapeutic use , Male , Mice , Models, Molecular , Quantitative Structure-Activity Relationship , XenopusABSTRACT
Photodynamic therapy (PDT) and vascular-disrupting agents (VDA) each have their advantages in the treatment of solid tumors, but also present drawbacks. In PDT, hypoxia at the center of the tumor limits conversion of molecular oxygen into singlet oxygen, while VDAs are deficient at affecting the rim of the tumor. A phthalocyanine-chalcone conjugate combining the VDA properties of chalcones with the PDT properties of phthalocyanines was designed to address these deficiencies. Its vascular targeting, photophysical, photochemical, photodynamic activities are reported herein.
Subject(s)
Chalcone/chemistry , Indoles/chemistry , Photochemotherapy/methods , Photosensitizing Agents/chemistry , Photosensitizing Agents/pharmacology , Cell Line , Cell Movement/drug effects , Chalcone/pharmacology , Humans , Indoles/pharmacology , Isoindoles , Molecular Structure , Photosensitizing Agents/chemical synthesis , Singlet Oxygen/metabolismABSTRACT
Disrupting the interaction between the PDZ protein PSD-95 and the C-terminal domain of the 5-HT2A serotonin receptor has been shown to reduce hyperalgesia in a rodent model of neuropathic pain. Here, we designed and synthesized PDZ ligands capable of binding to the first PDZ domain (PDZ1) of the PSD-95 protein and evaluated their biological activity in vitro and in vivo. A series of substituted indoles was identified by docking simulations, and six novel analogues were synthesized. Three analogues displayed strong interactions with the first PDZ domain (PDZ1) of PDZ-95 in (1)H-(15)N heteronuclear single-quantum coherence (HSQC) experiments and two of them were able to inhibit the interaction between PSD-95 and the 5-HT2A receptor in vitro. We identified compound 8b as the analogue able to significantly suppress mechanical hyperalgesia in an experimental model of traumatic neuropathic pain in the rat. This effect was suppressed by the coadministration of the 5-HT2A receptor antagonist M100907, consistent with an inhibitory effect upon 5-HT2A receptor/PSD-95 interaction. Finally, we determined an NMR-restraint driven model structure for the PSD95 PDZ1/8b complex, which confirms that indole 8b binds to the putative PDZ-ligand binding site.
Subject(s)
Analgesics/chemistry , Hyperalgesia/drug therapy , PDZ Domains , Receptor, Serotonin, 5-HT2A/metabolism , Serotonin 5-HT2 Receptor Antagonists/chemistry , Amino Acid Sequence , Analgesics/chemical synthesis , Analgesics/pharmacology , Animals , Computer Simulation , Disease Models, Animal , Ligands , Magnetic Resonance Spectroscopy , Molecular Structure , Rats , Serotonin 5-HT2 Receptor Antagonists/chemical synthesis , Serotonin 5-HT2 Receptor Antagonists/pharmacology , Structure-Activity RelationshipABSTRACT
Disrupting the interaction between the PDZ protein, PSD-95, and its target ligands (such as the glutamate NMDA receptor or the serotonin 5-HT2A receptor) was found to reduce hyperalgesia in various models of neuropathic pain. Here, we set out to identify lead molecules which would interact with PSD-95, and hence, would potentially display analgesic activity. We describe the virtual screening of the Asinex and Cambridge databases which together contain almost one million molecules. Using three successive docking filters and visual inspection, we identified three structural classes of molecules and synthesized a potential lead compound from each class. The binding of the molecules with the PDZ domains of PSD-95 was assessed by (1)H-(15)N HSQC NMR experiments. The analgesic activity of the best ligand, quinoline 2, was evaluated in vivo in a model of neuropathic pain and showed promising results.
Subject(s)
Analgesics/chemistry , Drug Design , Ligands , Nerve Tissue Proteins/chemistry , Analgesics/chemical synthesis , Analgesics/therapeutic use , Animals , Binding Sites , Molecular Docking Simulation , Nerve Tissue Proteins/metabolism , Neuralgia/drug therapy , PDZ Domains , Quinolines/chemistry , Rats , Receptor, Serotonin, 5-HT2A/chemistry , Receptor, Serotonin, 5-HT2A/metabolism , Receptors, N-Methyl-D-Aspartate/antagonists & inhibitors , Receptors, N-Methyl-D-Aspartate/metabolism , SAP90-PSD95 Associated ProteinsABSTRACT
A phthalocyanine-chalcone conjugate has been designed to combine the vascular disrupting effect of chalcones with the photodynamic effect of phthalocyanines. This potential dual photodynamic and antiangiogenic agent was obtained by the condensation of a tetrahydroxylated non-peripherally substituted Zn(ii) phthalocyanine with an amino chalcone converted into the corresponding activated isocyanate. The conjugate was fully characterized.
Subject(s)
Angiogenesis Inhibitors/chemical synthesis , Drug Design , Indoles/chemistry , Organometallic Compounds/chemistry , Photosensitizing Agents/chemical synthesis , Angiogenesis Inhibitors/chemistry , Chalcone/analogs & derivatives , Chalcone/chemistry , Isoindoles , Molecular Structure , Photosensitizing Agents/chemistry , Stereoisomerism , Zinc CompoundsABSTRACT
We designed bidentate ligands to target PDZ domains through two binding sites: site S0, delimited by the GLGF loop, and site S1, a zone situated around loop ß(B)/ß(C). A molecular docking study allowed us to design a generic S0 binder, to which was attached a variable size linker, itself linked to an amino acid aimed to interact with the S1 site of PDZ domains. A series of 15 novel bidentate ligands was prepared in 6-11 steps in good overall yield (24-43%). Some of these ligands showed an inhibitory activity against serotonin 5-HT2A receptor/PSD-95 interaction. This was assessed by pull-down assay using a synthetic decapeptide corresponding to the C-terminal residues of the receptor as a bait.
Subject(s)
Drug Design , Intracellular Signaling Peptides and Proteins/antagonists & inhibitors , Membrane Proteins/antagonists & inhibitors , PDZ Domains/drug effects , Serotonin 5-HT2 Receptor Agonists/pharmacology , Binding Sites/drug effects , Computational Biology , Disks Large Homolog 4 Protein , Intracellular Signaling Peptides and Proteins/chemistry , Ligands , Membrane Proteins/chemistry , Models, Molecular , Molecular Conformation , Receptor, Serotonin, 5-HT2A/chemistry , Receptor, Serotonin, 5-HT2A/metabolism , Serotonin 5-HT2 Receptor Agonists/chemical synthesis , Serotonin 5-HT2 Receptor Agonists/chemistry , Stereoisomerism , Structure-Activity RelationshipABSTRACT
We synthesized small organic molecules designed as PDZ ligands. These indole-based compounds were evaluated for their interaction with the PDZ1 domain of the post-synaptic density 95 (PSD-95) protein. Three molecules were found to interact with the targeted PDZ protein by NMR. One of them showed chemical shift perturbations closely related to the natural ligands.
Subject(s)
Guanylate Kinases/chemistry , Indoles/chemistry , Magnetic Resonance Spectroscopy , Membrane Proteins/chemistry , Models, Molecular , Animals , Disks Large Homolog 4 Protein , Hydrogen Bonding , Indoles/chemical synthesis , Mice , Protein Structure, TertiaryABSTRACT
Various methoxy- and hydroxy-substituted dibenz[c,e]oxepines were prepared via the copper(I)-induced coupling of ether-tethered arylstannanes or the dehydrative cyclisation of 1,1'-biphenyl-2,2'-dimethanols, assembled using the Ullmann cross-coupling of ortho-bromoaryl carbonyl compounds. The dibenzoxepines were screened for their ability to inhibit tubulin polymerisation and the in vitro growth of K562 human chronic myelogenous leukemia cells. The most active was 5,7-dihydro-3,9,10,11-tetramethoxydibenz[c,e]oxepin-4-ol, whose tubulin inhibitory and cytotoxicity (IC(50)) values were 1 µM and 40 nM, respectively.
Subject(s)
Dibenzoxepins/chemistry , Neovascularization, Pathologic , Tubulin/chemistry , Dibenzoxepins/metabolism , Dibenzoxepins/pharmacology , Humans , K562 Cells , Models, Molecular , Protein Binding , Tubulin/metabolismABSTRACT
A series of cis-restricted 1,4- and 1,5-disubstituted 1,2,3-triazole analogs of combretastatin A-4 (1) have been prepared. Cytotoxicity and tubulin inhibition studies showed that 2-methoxy-5-((5-(3,4,5-trimethoxyphenyl)-1H-1,2,3-triazol-1-yl)methyl)aniline (5e) and 2-methoxy-5-(1-(3,4,5-trimethoxybenzyl)-1H-1,2,3-triazol-5-yl)aniline (6e) were two of the most active compounds. Molecular modeling studies revealed that the N-2 and N-3 atoms in the triazole rings in 5e and 6e did not form hydrogen bonds with the amino acids in the anticipated pharmacophore.
Subject(s)
Aniline Compounds/chemistry , Microtubules/chemistry , Stilbenes/chemistry , Triazoles/chemistry , Tubulin Modulators/chemistry , Aniline Compounds/chemical synthesis , Aniline Compounds/toxicity , Binding Sites , Cell Line, Tumor , Computer Simulation , Humans , Microtubules/metabolism , Protein Structure, Tertiary , Stilbenes/chemical synthesis , Stilbenes/toxicity , Triazoles/chemical synthesis , Triazoles/toxicity , Tubulin Modulators/chemical synthesis , Tubulin Modulators/toxicityABSTRACT
The alpha-methyl chalcone SD400 is a potent inhibitor of tubulin assembly and possesses potent anticancer activity. Various chalcone analogues were synthesized and evaluated for their cell growth inhibitory properties against the K562 human chronic myelogenous leukemia cell line (SD400, IC(50) 0.21nM; combretastatin A4 CA4, IC(50) 2.0nM). Cell cycle analysis by flow cytometry indicated that these agents are antimitotic (SD400, 83% of the cells are in G(2)/M phase; CA4 90%). They inhibit tubulin assembly at low concentration (SD400, IC(50) 0.46microM; CA4, 0.10microM) and compete with [(3)H]colchicine for binding to tubulin (8% [(3)H]colchicine remained bound to tubulin after competition with SD400 or CA4). Upon treatment with SD400, remarkable cell shape changes were elicited in HUVEC cells, consistent with vasculature damaging activity.
Subject(s)
Antineoplastic Agents/chemical synthesis , Bibenzyls/pharmacology , Cell Proliferation/drug effects , Chalcones/pharmacology , Microtubules/drug effects , Neovascularization, Physiologic/drug effects , Tubulin Modulators/chemical synthesis , Antineoplastic Agents/chemistry , Antineoplastic Agents/pharmacology , Bibenzyls/chemical synthesis , Bibenzyls/chemistry , Cell Line, Tumor , Chalcones/chemical synthesis , Chalcones/chemistry , Drug Evaluation, Preclinical , Drug Screening Assays, Antitumor , Humans , K562 Cells , Microtubules/metabolism , Structure-Activity Relationship , Tubulin Modulators/chemistry , Tubulin Modulators/pharmacologyABSTRACT
Tubulin is an important molecular target in cancer chemotherapy. Antimitotic agents able to bind to the protein are currently under study, commonly used in the clinic to treat a variety of cancers and/or exploited as probes to investigate the protein's structure and function. Here we report the binding modes for a series of colchicinoids, combretastatin A4 and chalcones established from docking studies carried out on the structure of tubulin in complex with colchicine. The proposed models, in agreement with published biochemical data, show that combretastatin A4 binds to the colchicine site of beta-tubulin and that chalcones assume an orientation similar to that of podophyllotoxin. The models can be used to design a new class of podophyllotoxin mimics, the alpha-aryl chalcones, capable of binding to the colchicine-binding site of beta-tubulin with higher affinity.
Subject(s)
Bibenzyls/pharmacology , Chalcones/chemistry , Chalcones/pharmacology , Microtubules/drug effects , Tubulin Modulators/pharmacology , Algorithms , Bibenzyls/chemical synthesis , Bibenzyls/chemistry , Binding Sites , Cell Line , Chalcones/chemical synthesis , Colchicine/analogs & derivatives , Colchicine/chemical synthesis , Colchicine/chemistry , Colchicine/pharmacology , Drug Discovery , Humans , Hydrocarbons, Cyclic/chemical synthesis , Hydrocarbons, Cyclic/chemistry , Hydrocarbons, Cyclic/pharmacology , Microtubules/metabolism , Structure-Activity Relationship , Tubulin Modulators/chemical synthesis , Tubulin Modulators/chemistryABSTRACT
To further pursue the antineoplastic leads offered by our isolation of trans-dihydronarciclasine (1a) and 7-deoxy-trans-dihydronarciclasine (1c) from two medicinal plant species of the Amaryllidaceae family, a practical palladium-catalyzed hydrogenation procedure was developed for the synthesis of these isocarbostyrils from narciclasine (2a) and 7-deoxynarciclasine (2c).
Subject(s)
Alkaloids/chemical synthesis , Alkaloids/pharmacology , Antineoplastic Agents, Phytogenic/chemical synthesis , Isoquinolines/chemical synthesis , Isoquinolines/pharmacology , Narcissus/chemistry , Plants, Medicinal/chemistry , Alkaloids/chemistry , Amaryllidaceae Alkaloids , Antineoplastic Agents, Phytogenic/chemistry , Antineoplastic Agents, Phytogenic/pharmacology , Catalysis , Isoquinolines/chemistry , Molecular Structure , Palladium/chemistry , Phenanthridines , StereoisomerismABSTRACT
The development of chalcones as antimitotic agents has led to the design of other analogues able to interact with tubulin and inhibit its assembly into microtubules. This activity has also been associated with their anti-vascular activity. This review focuses on the development of chalcones and related analogues as antimitotic agents.