Vyera Pharmaceuticals

Insomnia disorder is characterized by the difficulty in initiating or maintaining sleep and corresponding daytime dysfunction occurs in 10–20% of the population. Although the molecular underpinnings of sleep-onset insomnia remain unclear many large-scale genetic studies have been conducted pointing to UGT1A1 loci as some of the strongest risk variants. UGT1A1 codes for an enzyme that is responsible for glucuronidation of bilirubin and may a role in relationship between insomnia and bilirubin metabolism. In this study, we explored the association of bilirubin levels in insomnia patients as compared with controls and in association with UGT1A variants. Blood samples were obtained from adults (ages 18-70) with a diagnosis of insomnia participating in a clinical trial of 322 patients with primary using polysomnography (PSG) measures of sleep. Bilirubin was measured at baseline in the serum of the patients. Insomnia patients (n=322) at baseline had significantly lower levels of serum bilirubin when compared to healthy sleeping cohort as controls (n=273, p-value=1.75e-14), with mean values 0.32 MG/DL in cases and 0.51 controls. This effect was consistent when using a different set of controls (n=2841, p-value=8.09e-17, mean=0.44) and upon correction for covariates. Furthermore, we have run a genome wide association analysis using linear model to study variants associated with bilirubin levels on variants with MAF ≥0.01. One of the top variants associated with bilirubin at baseline in the insomnia cases was rs4148325 with a MAF of 0.36, where minor allele was associated with higher bilirubin levels (rs4148325 (β = 0.2 mg/dl per T allele increase; p = 3.82e-11)). We expanded the analysis with single gene tests which showed that UGT1A1 gene is significant especially for MAF>0.05 variants (SKAT p=0.009). The variant with highest OR in terms of cases/controls was rs1042640 a UTR3 variant with a MAF of 0.20. Hypobilirubinemia is most commonly seen in patients with metabolic dysfunction, which may lead to cardiovascular complications and possibly stroke. The circadian clock was shown to protect against the neurotoxic consequences of increased bilirubin by regulating bilirubin detoxification. Further studies are warranted in order the further understand the aberrant levels on bilirubin in insomnia patients, and fundamentally their cause of an effect aspects.

Lenacapavir (LEN) is a novel, first-in-class, multistage, selective inhibitor of human immunodeficiency virus type 1 (HIV-1) capsid function recently approved for the treatment of HIV-1 infection in heavily treatment-experienced adults with multidrug-resistant HIV-1 infection. The purpose of this multicohort study was to evaluate the pharmacokinetics, metabolism, excretion, safety, and tolerability of LEN following a single intravenous (IV) infusion of 10 mg LEN or 20 mg [14C]LEN in healthy participants. Twenty-one healthy adult participants were enrolled into the study and received either a single IV dose of 10 mg LEN (n = 8 active, n = 3 placebo; cohort 1) or a single IV dose of 20 mg [14C]LEN containing 200 µCi (n = 10; cohort 2). Blood, urine, and feces samples (when applicable) were collected after dosing, and radioactivity (cohort 2) was assessed using liquid scintillation counting in both plasma and excreta. LEN in plasma was quantified by liquid chromatography (LC) tandem mass spectroscopy (MS/MS) method bioanalysis. Metabolite profiling in plasma and excreta were performed using LC-fraction collect (FC)-high-resolution MS and LC-FC-accelerator mass spectrometry in plasma. Between the 10 mg and 20 mg doses of LEN, the observed plasma exposure of LEN doubled, while the elimination half-life was similar. Following administration of 20 mg [14C]LEN (200 µCi), the mean cumulative recovery of [14C] radioactivity was 75.9

Pancreatic ductal adenocarcinoma (PDAC) remains one of the deadliest solid cancers and thus identifying more effective therapies is a major unmet need. In this study we characterized the super enhancer (SE) landscape of human PDAC to identify novel, potentially targetable, drivers of the disease. Our analysis revealed that MICAL2 is a super enhancer-associated gene in human PDAC. MICAL2 is a flavin monooxygenase that induces actin depolymerization and indirectly promotes SRF transcription by modulating the availability of serum response factor coactivators myocardin related transcription factors (MRTF-A and MRTF-B). We found that MICAL2 is overexpressed in PDAC and correlates with poor patient prognosis. Transcriptional analysis revealed that MICAL2 upregulates KRAS and EMT signaling pathways, contributing to tumor growth and metastasis. In loss and gain of function experiments in human and mouse PDAC cells, we observed that MICAL2 promotes both ERK1/2 and AKT activation. Consistent with its role in actin depolymerization and KRAS signaling, loss of MICAL2 expression also inhibited macropinocytosis. Through in vitro phenotypic analyses, we show that MICAL2, MRTF-A and MRTF-B influence PDAC cell proliferation, migration and promote cell cycle progression. Importantly, we demonstrate that MICAL2 is essential for in vivo tumor growth and metastasis. Interestingly, we find that MRTF-B, but not MRTF-A, phenocopies MICAL2-driven phenotypes in vivo . This study highlights the multiple ways in which MICAL2 impacts PDAC biology and suggests that its inhibition may impede PDAC progression. Our results provide a foundation for future investigations into the role of MICAL2 in PDAC and its potential as a target for therapeutic intervention.

Introduction A growing body of evidence suggests a role for HCN (hyperpolarization-activated cyclic nucleotide-gated) channels in regulation of sleep. We first tested effects of three HCN channel blockers (Corlanor, Zatebradine Hydrochloride and ZD7288) on sleep/wake behavior in wild type zebrafish compared to DMSO control. Then, to more definitively investigate roles of HCN channels, we knocked out genes encoding for HCN channel subunits in zebrafish by utilizing CRISPR/Cas9 technique and we assessed sleep/wake behavior in the HCN channel mutants. Methods The compounds were tested at six concentrations varying between 0.1-30 ◻M (.i.e., 0.1 μM, 0.3 μM, 1.0 μM, 4.5 μM, 10 μM and 30 μM) in zebrafish larvae in our drug screening study. Control groups were administered DMSO. In CRISPR screening, single cell-stage zebrafish embryos were injected with preformed ribonucleoprotein complexes containing Cas9 protein and two crRNAs targeting each homologous gene of human HCN1, HCN2, HCN3, and HCN4. Control groups were negative control crRNA injected embryos. Sleep phenotyping was performed by placing individual larva into the wells of 96 well plates and by recording their activity via commercially available video monitoring equipment. Results Drug screening study showed shorter latency to sleep at 0.1 μM dose of Ivabradine, moderate reductions in average activity at 30 μM dose of Zatebradine Hydrochloride, and increased sleep at 4.5 μM dose of ZD7288. Our CRISPR/Cas9 screening resulted in decreased activity in hcn1 crispants, fewer sleep bouts in daytime in hcn2 crispants, and no difference in sleep parameters in hcn3 crispants. hcn4 crispants had shorter sleep in both day and nighttime, hyperactivity and shorter sleep bouts at night and fewer sleep bouts in daytime. Conclusion Blocking HCN channels decreased wakefulness in our drug screening study. Genetically targeting hcn1 and hcn2 genes resulted in the same direction with the results of drug screening study and knocking out hcn3 didn’t change sleep parameters. Change in sleep parameters were in the opposite direction in hcn4 crispants. HCN channel blockers might be selecting HCN1 and HCN2 however further work is required to confirm this suggestion. Support (if any)