RESUMO
Chordoma is a devastating rare cancer that affects one in a million people. With a mean-survival of just 6 years and no approved medicines, the primary treatments are surgery and radiation. In order to speed new medicines to chordoma patients, a drug repurposing strategy represents an attractive approach. Drugs that have already advanced through human clinical safety trials have the potential to be approved more quickly than de novo discovered medicines on new targets. We have taken two strategies to enable this: (1) generated and validated machine learning models of chordoma inhibition and screened compounds of interest in vitro. (2) Tested combinations of approved kinase inhibitors already being individually evaluated for chordoma. Several published studies of compounds screened against chordoma cell lines were used to generate Bayesian Machine learning models which were then used to score compounds selected from the NIH NCATS industry-provided assets. Out of these compounds, the mTOR inhibitor AZD2014, was the most potent against chordoma cell lines (IC50 0.35 µM U-CH1 and 0.61 µM U-CH2). Several studies have shown the importance of the mTOR signaling pathway in chordoma and suggest it as a promising avenue for targeted therapy. Additionally, two currently FDA approved drugs, afatinib and palbociclib (EGFR and CDK4/6 inhibitors, respectively) demonstrated synergy in vitro (CI50 = 0.43) while AZD2014 and afatanib also showed synergy (CI50 = 0.41) against a chordoma cell in vitro. These findings may be of interest clinically, and this in vitro- and in silico approach could also be applied to other rare cancers.
Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica/farmacologia , Cordoma/tratamento farmacológico , Reposicionamento de Medicamentos , Aprendizado de Máquina , Protocolos de Quimioterapia Combinada Antineoplásica/química , Benzamidas/agonistas , Benzamidas/farmacologia , Linhagem Celular Tumoral , Cordoma/metabolismo , Cordoma/patologia , Humanos , Morfolinas/agonistas , Morfolinas/farmacologia , Proteínas de Neoplasias/antagonistas & inibidores , Proteínas de Neoplasias/metabolismo , Piperazinas/agonistas , Piperazinas/farmacologia , Piridinas/agonistas , Piridinas/farmacologia , Pirimidinas/agonistas , Pirimidinas/farmacologiaAssuntos
Anilidas/administração & dosagem , Antineoplásicos/administração & dosagem , Síndrome do Nevo Basocelular/tratamento farmacológico , Piridinas/administração & dosagem , Adulto , Anilidas/efeitos adversos , Anilidas/agonistas , Antineoplásicos/efeitos adversos , Síndrome do Nevo Basocelular/patologia , Esquema de Medicação , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Piridinas/efeitos adversos , Piridinas/agonistas , Neoplasias Cutâneas/tratamento farmacológico , Neoplasias Cutâneas/patologia , Resultado do TratamentoRESUMO
BACKGROUND: Over recent years, the 5-hydroxytryptamine6 (5-HT6) receptor has emerged as a promising molecular target which interacts with several central nervous system acting drugs. In animal models, both agonists and antagonists of this receptor exhibit equivalent potency and efficacy as potential antidepressants, anxiolytics and anti-obesity or anti-dementia drugs. EMD386088 (5-chloro-2-methyl-3-(1,2,3,6-tetrahydro-4-pyridinyl)-1H-indole hydrochloride) has been described as a high affinity 5-HT6 receptor ligand with a full agonist activity and with moderate affinity for 5-HT3 sites. METHODS: We have extended these data by broadening its profile for other, not yet tested, monoaminergic, GABA(A), opioid µ receptors and serotonin transporter (SERT) and we have conducted functional in vitro assays; i.e., measurement of cAMP by homogeneous TR-FRET immunoassay and HTRF method made by CEREP as well as aequorin-based calcium flux assay. RESULTS: In two in vitro models based on cAMP formation, maximal efficacy values for EMD386088 were 65 and 31%, for in house and CEREP experiments, respectively. In a model based on calcium response, the studied compound showed 46% of maximal serotonin (5-HT) signal. EMD386088 antagonizes 5-HT response in increasing concentrations from 10(-9) to 10(-6) M. CONCLUSIONS: The present in vitro findings confirm that EMD386088 is a selective 5-HT6 receptor ligand with moderate affinity for 5-HT3 sites only and it behaves as a potent partial agonist of 5-HT6 receptor with varying levels of agonist intrinsic activity, depending on a method employed. In view of these results, caution is recommended in the interpretation of pharmacological in vivo studies with EMD386088.
Assuntos
Agonismo Parcial de Drogas , Indóis/agonistas , Indóis/antagonistas & inibidores , Piridinas/agonistas , Piridinas/antagonistas & inibidores , Receptores de Serotonina/efeitos dos fármacos , Agonistas do Receptor de Serotonina/farmacologia , Animais , Células CHO , Cálcio/metabolismo , Células Cultivadas , Cricetulus , AMP Cíclico/metabolismo , Humanos , Ensaio Radioligante , Serotonina/farmacologia , Antagonistas da Serotonina/farmacologiaRESUMO
Radial artery (RA) vasospasm remains a potential cause of early graft failure after coronary artery bypass graft surgery, despite pretreatment with alpha-adrenergic or calcium channel blockers. Our aim was to investigate the mechanism of the vasorelaxant effects of Rho-kinase inhibitors (Y-27632 and fasudil) on the human RA. Segments were obtained from 30 patients undergoing coronary artery bypass graft and were divided into 3-4 mm vascular rings. The rings were stimulated with 10(-5) mol/L phenylephrine (PE) by using the isolated tissue bath technique and were relaxed with 10(-6) mol/L acetylcholine. Relaxation responses were recorded for Y-27632 (10(-9)-10(-4) mol/L), fasudil (10(-9)-10(-4) mol/L), and sodium nitroprusside (SNP) (10(-9)-10(-5) mol/L). Y-27632 and fasudil relaxation responses were repeated in either N(G)-nitro-L-arginine (L-NNA), which is a specific endothelial nitric oxide synthase inhibitor, or 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ), which is a guanylate cyclase inhibitor. SNP relaxation responses were repeated in 10(-8) mol/L Y-27632 and 10(-8) mol/L fasudil. Y-27632 and fasudil caused concentration-dependent vasorelaxation in RA rings precontracted with PE, and maximal relaxation (100%) was recorded at the highest concentration used (10(-4) mol/L). The vasorelaxant effects of Y-27632 and fasudil were significantly reduced in the presence of L-NNA and ODQ, and the pD2 values of Y-27632 and fasudil were not changed. The vasorelaxant effects of SNP were significantly increased in the presence of Y-27632 and fasudil, and the pD(2) values of SNP were not changed. These findings indicate that Y-27632 and fasudil caused concentration-dependent vasorelaxation in the RA rings. Because this effect was decreased in a dose-dependent manner by L-NNA and ODQ, the relaxant effects of Y-27632 and fasudil could be due to stimulation by nitric oxide that is being released. Rho-kinase inhibitors may have an important role in preventing vasospasm in arterial grafts used for coronary artery surgery.
Assuntos
1-(5-Isoquinolinasulfonil)-2-Metilpiperazina/análogos & derivados , Amidas/farmacologia , Inibidores de Proteínas Quinases/agonistas , Inibidores de Proteínas Quinases/farmacologia , Piridinas/farmacologia , Artéria Radial/efeitos dos fármacos , Vasoconstrição/efeitos dos fármacos , Quinases Associadas a rho/antagonistas & inibidores , 1-(5-Isoquinolinasulfonil)-2-Metilpiperazina/agonistas , 1-(5-Isoquinolinasulfonil)-2-Metilpiperazina/farmacologia , Idoso , Amidas/agonistas , Constrição Patológica/enzimologia , Constrição Patológica/prevenção & controle , Relação Dose-Resposta a Droga , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Nitroarginina/farmacologia , Nitroprussiato/farmacologia , Oxidiazóis/farmacologia , Piridinas/agonistas , Quinoxalinas/farmacologia , Artéria Radial/enzimologia , Receptores de Detecção de Cálcio/agonistas , Receptores de Detecção de Cálcio/fisiologia , Vasoconstrição/fisiologia , Vasodilatação/efeitos dos fármacos , Vasodilatação/fisiologia , Quinases Associadas a rho/fisiologiaRESUMO
The electrophysiological actions of various neonicotinoids, including substituted benzyl derivatives, against recombinant Drosophila SAD/chicken beta2 hybrid nicotinic acetylcholine receptor (nAChR) were measured to analyze the relationships between the in vivo (insecticidal) and in vitro (binding and agonist) activities. Most of the neonicotinoids tested were capable of inducing inward currents by activating the hybrid nAChRs expressed in Xenopus laevis oocytes, whereas some compounds had no agonist activity and only blocked the acetylcholine-induced currents. Variations in the agonist activity were well correlated with those in the binding potency evaluated using [3H]imidacloprid as well as insecticidal activities.