ABSTRACT
Activation of soluble guanylate cyclase (sGC) to restore cyclic guanosine monophosphate (cGMP) and improve functionality of nitric oxide (NO) pathways impaired by oxidative stress is a potential treatment of diabetic and chronic kidney disease. We report the pharmacology of BI 685509, a novel, orally active small molecule sGC activator with disease-modifying potential. BI 685509 and human sGC α1/ß1 heterodimer containing a reduced heme group produced concentration-dependent increases in cGMP that were elevated modestly by NO, whereas heme-free sGC and BI 685509 greatly enhanced cGMP with no effect of NO. BI 685509 increased cGMP in human and rat platelet-rich plasma treated with the heme-oxidant ODQ; respective EC50 values were 467 nM and 304 nM. In conscious telemetry-instrumented rats, BI 685509 did not affect mean arterial pressure (MAP) or heart rate (HR) at 3 and 10 mg/kg (p.o.), whereas 30 mg/kg decreased MAP and increased HR. Ten days of BI 685509 at supratherapeutic doses (60 or 100 mg/kg p.o., daily) attenuated MAP and HR responses to a single 100 mg/kg challenge. In the ZSF1 rat model, BI 685509 (1, 3, 10, and 30 mg/kg per day, daily) coadministered with enalapril (3 mg/kg per day) dose-dependently reduced proteinuria and incidence of glomerular sclerosis; MAP was modestly reduced at the higher doses versus enalapril. In the 7-day rat unilateral ureteral obstruction model, BI 685509 dose-dependently reduced tubulointerstitial fibrosis (P < 0.05 at 30 mg/kg). In conclusion, BI 685509 is a potent, orally bioavailable sGC activator with clear renal protection and antifibrotic activity in preclinical models of kidney injury and disease. SIGNIFICANCE STATEMENT: BI 685509 is a novel small soluble guanylate cyclase (sGC) molecule activator that exhibits an in vitro profile consistent with that of an sGC activator. BI 685509 reduced proteinuria and glomerulosclerosis in the ZSF1 rat, a model of diabetic kidney disease (DKD), and reduced tubulointerstitial fibrosis in a rat 7-day unilateral ureteral obstruction model. Thus, BI 685509 is a promising new therapeutic agent and is currently in phase II clinical trials for chronic kidney disease and DKD.
Subject(s)
Renal Insufficiency, Chronic , Ureteral Obstruction , Rats , Humans , Animals , Soluble Guanylyl Cyclase/metabolism , Guanylate Cyclase/metabolism , Ureteral Obstruction/pathology , Kidney/metabolism , Disease Progression , Proteinuria/drug therapy , Fibrosis , Enalapril/therapeutic use , Nitric Oxide/metabolism , Cyclic GMP/metabolismABSTRACT
Microbial-dependent trimethylamine (TMA) generation from dietary precursors such as choline was recently linked to cardiovascular diseases (CVDs) as well as chronic kidney disease (CKD). Inhibition of TMA-generating enzymes in gut bacteria would be an innovative approach to treat these diseases. The potential to accurately quantify secreted TMA levels highlights the capacity of mass spectrometry (MS) for tracking microbial TMA-lyase activity. However, high-throughput screening (HTS) by conventional MS instrumentation is hampered by limited sample throughput. Recent advancement in liquid handling and instrumentation of matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) MS provides an HTS-compatible MS technology. The deciphering of enzymatic reactions using this label-free readout has been successfully applied but has thus far been limited to peptide/protein-centric activity assays. Here, we demonstrate the versatile applicability of MALDI-TOF by tracking a small molecule within a highly complex sample background. The key to success for this concept was chemical derivatization of the target molecule enabling quantitative assessment of microbial TMA formation. Further, its potential was demonstrated in a side-by-side comparison to RapidFire-MS in a primary screen and subsequent dose-response experiments. Overall, the established assay enables the screening for microbial TMA-lyase inhibitors and serves as a proof of concept for the applicability of MALDI-TOF for demanding assay concepts per se.
Subject(s)
Drug Discovery/methods , Enzyme Inhibitors/pharmacology , High-Throughput Screening Assays , Lyases/antagonists & inhibitors , Methylamines/metabolism , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , HumansABSTRACT
A new class of chymase inhibitor featuring a benzimidazolone core with an acid side chain and a P(1) hydrophobic moiety is described. Incubation of the lead compound with GSH resulted in the formation of a GSH conjugate on the benzothiophene P(1) moiety. Replacement of the benzothiophene with different heterocyclic systems such as indoles and benzoisothiazole is feasible. Among the P(1) replacements, benzoisothiazole prevents the formation of GSH conjugate and an in silico analysis of oxidative potentials agreed with the experimental outcome.
Subject(s)
Benzimidazoles/chemistry , Chymases/antagonists & inhibitors , Protease Inhibitors/chemistry , Benzimidazoles/metabolism , Benzimidazoles/pharmacology , Binding Sites , Chymases/metabolism , Crystallography, X-Ray , Humans , Hydrophobic and Hydrophilic Interactions , Oxidation-Reduction , Protease Inhibitors/chemical synthesis , Protease Inhibitors/pharmacology , Protein Structure, Tertiary , Structure-Activity RelationshipABSTRACT
As doenças crônico-degenerativas são doenças progressivas e que interferem na qualidade de vida de seus portadores. Tendo em vista esse aspecto e o fato de ser a humanização das relações e do cuidado ao ser humano uma preocupação de profissionais desaúde e de cuidadores, este estudo tem como objetivo esclarecer dúvidas para que se possa repensar as relações e os valores éticos no processo do cuidar. Além da eficiência técnico-científica, os caminhos apontam também para a prática da sensibilidade e da solidariedade humana.
Chronic-degenerative diseases are gradual diseases that affect the quality of life of sick people. In view of this aspect and the goal of humanization of relationships and care of people as concerns of health professionals and carers, this study aims to dispel doubts so that one can rethink the ethical relations and values in the process of taking care. Beyond technical-scientific efficiency, there are also ways for practicing sensitivity and solidarity regarding human beings.
Las enfermedades crónico-degenerativas son enfermedades graduales que afectan a la calidad de vida de la gente enferma. Debido a este aspecto y a la meta de la humanización de las relaciones y del cuidado de la gente como preocupaciones de los profesionales de salud y de los cuidadores, este estudio busca disipar dudas de modo que uno pueda repensar las relaciones y los valores éticos en elcurso del cuidado. Más allá de la eficacia técnico-científica, hay también maneras de practicar la sensibilidad y la solidaridad respecto a los seres humanos.
