In vitro and in vivo characterization of potent antileishmanial methionine aminopeptidase 1 inhibitors

Leishmania major is the causative agent of cutaneous leishmaniasis (CL). No human vaccine is available for CL, and current drug regimens present several drawbacks, such as emerging resistance, severe toxicity, medium effectiveness, and/or high cost. Thus, the need for better treatment options against CL is a priority. In the present study, we validate the enzyme methionine aminopeptidase 1 of L. major (MetAP1Lm), a metalloprotease that catalyzes the removal of N-terminal methionine from peptides and proteins, as a chemotherapeutic target against CL infection. The in vitro antileishmanial activities of eight novel MetAP1 inhibitors (OJT001 to OJT008) were investigated. Three compounds, OJT006, OJT007, and OJT008, demonstrated potent antiproliferative effects in macrophages infected with L. major amastigotes and promastigotes at submicromolar concentrations, with no cytotoxicity against host cells. Importantly, the leishmanicidal effect in transgenic L. major promastigotes overexpressing MetAP1Lm was diminished by almost 10-fold in comparison to the effect in wild-type promastigotes. Furthermore, the in vivo activities of OJT006, OJT007, and OJT008 were investigated in L. major-infected BALB/c mice. In comparison to the footpad parasite load in the control group, OJT008 decreased the footpad parasite load significantly, by 86%, and exhibited no toxicity in treated mice. We propose MetAP1 inhibitor OJT008 as a potential chemotherapeutic candidate against CL infection caused by L. major infection

HIV-TB Coinfection among 57 Million Pregnant Women, Obstetric Complications, Alcohol Use, Drug Abuse, and Depression

Objective. HIV and tuberculosis represent diseases of major public health importance worldwide. Very little is known about HIV-TB coinfection among pregnant women, especially from industrialized settings. In this study, we examined the association between TB, HIV, and HIV-TB coinfection among pregnant mothers and obstetric complications, alcohol use, drug abuse, and depression. Method. We examined inpatient hospital discharges in the United States from January 1, 2002, through December 31, 2014. We employed multivariable survey logistic regression to generate adjusted estimates for the association between infection status and study outcomes. Results. We analyzed approximately 57 million records of pregnant women and their delivery information. HIV-TB coinfection was associated with the highest risks for several obstetric complications, alcohol use, and drug abuse. The risk for alcohol abuse was more than twice as high among HIV-monoinfected as compared to TB-monoinfected mothers. That risk gap more than doubled with HIV-TB coinfection. Both HIV-monoinfected and HIV-TB coinfected mothers experienced similarly increased risks for depression. Conclusions. Mothers with HIV-TB coinfection experienced relatively heightened risks for obstetric complications, alcohol use, and drug abuse. The findings of this study underscore the importance of augmenting and enhancing social and structural support systems for HIV-TB coinfected pregnant women

Trends in Appendicitis Among Pregnant Women, the Risk for Cardiac Arrest, and Maternal–Fetal Mortality

Background: Appendicitis is the most common extra-uterine surgical emergency requiring immediate intervention during pregnancy. However, risks for mortality and morbidity among pregnant women with appendicitis remain poorly understood. This study was conducted to determine the temporal trends of appendicitis in pregnant women, and to calculate the risk of maternal–fetal mortality and near-miss marker (i.e., cardiac arrest) among pregnant women in general, and by race/ethnicity. Methods: We conducted this retrospective study using data from the Nationwide Inpatient Sample (NIS) from January 1, 2002, through December 31, 2015. Joinpoint regression was used to estimate and describe temporal changes in the rates of all and acute appendicitis during the 14-year study period. We also estimated the risk of cardiac arrest, maternal, and fetal mortality among mothers of various racial/ethnic groups with a diagnosis of acute appendicitis. Within each group, patients without acute appendicitis were the referent category. Results and conclusions: Out of the 58 million pregnancy hospitalizations during the study period, 63,145 cases (10.74 per 10,000 hospitalizations) were for acute appendicitis. There was a 5% decline (95% CI: − 5.1, − 5.0) in the rate of appendicitis hospitalizations over the period of the study. After adjusting for covariates, pregnant mothers with acute appendicitis had increased likelihood when compared to those without acute appendicitis to suffer fetal loss (OR: 2.05, 95% CI: 1.85–2.28) and nearly fivefold increase for inpatient maternal death. In conclusion, appendicitis during pregnancy remains an important cause of in-hospital maternal–fetal mortality overall and regardless of race/ethnicity

Discovery of Clioquinol and analogues as novel inhibitors of Severe Acute Respiratory Syndrome Coronavirus 2 infection, ACE2 and ACE2 - Spike protein interaction in vitro

Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), the etiological agent for coronavirus disease 2019 (COVID-19), has resulted in an ongoing pandemic. Presently, there are no clinically approved drugs for COVID-19. Hence, there is an urgent need to accelerate the development of effective antivirals. Herein, we discovered Clioquinol (5-chloro-7-iodo-8-quinolinol (CLQ)), a Food and Drug Administration (FDA) approved drug, and two of its analogues (7-bromo-5-chloro-8-hydroxyquinoline (CLBQ14); and 5, 7-Dichloro-8-hydroxyquinoline (CLCQ)) as potent inhibitors of SARS-CoV-2 infection-induced cytopathic effect in vitro. In addition, all three compounds showed potent anti-exopeptidase activity against recombinant human angiotensin-converting enzyme 2 (rhACE2) and inhibited the binding of rhACE2 with SARS-CoV-2 Spike (RBD) protein. CLQ displayed the highest potency in the low micromolar range, with its antiviral activity showing a strong correlation with inhibition of rhACE2 and rhACE2-RBD interaction. Altogether, our findings provide a new mode of action and molecular target for CLQ and validates this pharmacophore as a promising lead series for the clinical development of potential therapeutics for COVID-19

A simple, sensitive and reliable LC-MS/MS method for the determination of 7-bromo-5-chloroquinolin-8-ol (CLBQ14), a potent and selective inhibitor of methionine aminopeptidases: Application to pharmacokinetic studies

CLBQ14 is an 8-hydroxyquinoline analogue that inhibits methionine aminopeptidase (MetAP), an enzyme responsible for the post-translational modification of several proteins and polypeptides. MetAP has been validated as druggable target for some infectious diseases, and its inhibitors have been investigated as potential therapeutic agents. In this study, we developed and validated a liquid chromatography tandem-mass spectrometry (LC-MS/MS) method for the quantification of CLBQ14 in solution, and in rat plasma and urine. This method was applied to the pharmacokinetic evaluation of CLBQ14 in adult male Sprague Dawley (SD) rats. Chromatographic separation was achieved using an ultra-high-performance liquid chromatography (UHPLC) system equipped with Waters XTerra MS C18 column (3.5 μm, 125 Å, 2.1 × 50 mm) using 0.1% formic acid in acetonitrile/water gradient system as mobile phase. Chromatographic analysis was performed with a 4000 QTRAP® mass spectrometer using MRM in positive mode for CLBQ14 transition [M + H]+ m/z 257.919 → m/z 151.005, and IS (clioquinol) transition [M + H]+ m/z 305.783 → m/z 178.917. CLBQ14 was extracted from plasma and urine samples by protein precipitation. The retention times for CLBQ14 and IS were 1.31 and 1.40 min respectively. The standard curves were linear for CLBQ14 concentration ranging from 1 to 1000 ng/mL. The intra-day and inter-day accuracy and precision were found to be within 15% of the nominal concentration. Extraction recoveries were \u3e96.3% and 96.6% from rat plasma and urine respectively, and there was no significant matrix effect from the biological matrices. CLBQ14 is stable in samples subjected to expected storage, preparation, and handling conditions. Pharmacokinetic studies revealed that CLBQ14 has a bi-exponential disposition in SD rats, is extensively distributed with a long plasma half-life and is eliminated primarily by liver metabolism

Validation of Methionine Aminopeptidase-1 as a Potential Chemotherapeutic Target for Leishmania major

p.p1 {margin: 0.0px 0.0px 0.0px 0.0px; font: 12.0px Garamond} span.s1 {font: 12.0px \u27Times New Roman\u27} span.s2 {font: 7.0px Garamond} Cutaneous leishmaniasis is a vector-borne disease caused by Leishmania major affecting millions of people throughout the world. It is an emerging concern in the United States due to military establishments in endemic countries. Treatment for the disease is highly toxic and the lack of vaccines emphasizes on the need for alternative drug treatments. Aminopeptidase inhibitors have shown promising results against malaria and tuberculosis. Methionine aminopeptadse-1 (MetAP1) catalyzes the removal of the N-terminal methionine residue from peptides and proteins. The human MetAP1 has low similarity with the Leishmania enzyme making it a potential chemotherapeutic target. The MetAP1 gene was amplified through PCR from L. major genomic DNA and cloned into the expression vector pRSET A. The expression and purification of the recombinant enzyme was performed and the biochemical characterization of the enzyme is underway. Transgenic promastigotes of L. major (Friedlin clone V1) expressing firefly luciferase were used to determine the antiparasitic activity of putative MetAP1 inhibitors. Eight compounds were tested and three of them showed great anti-leishmanial activity at 0.78μM for 72 hr. with 5-6% survival (IC50 = 125 nM). The enzymatic assay using the recombinant enzyme will be developed to determine the inhibitors’ specificity

The Global Alliance for Maternal and Child Health (GLAM): A Pioneer Organization for MCH Students

Currently, there is an insufficient representation of racial/ethnic minority groups in the maternal and child health (MCH) workforce. A student-run outreach organization, the Global Alliance for Maternal and Child Health (GLAM), seeks to address this disparity by increasing the representation of racial/ethnic minority groups in MCH workforce. Founded by students at Texas Southern University in Houston, Texas, United States, GLAM, seeks to establish productive alliances and create programs that would help improve the well-being of mothers, infants, and children locally, nationally, and internationally by engaging an active cadre of students passionate about MCH. Through community outreach and global engagement using evidence-based strategies, GLAM is committed to the elimination of health disparities plaguing the MCH population. Key words: • Maternal and Child Health • Pipeline program • Students-driven • Community outreach • Global engagement   Copyright © 2021 Harris et al. Published by Global Health and Education Projects, Inc. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0) which permits unrestricted use, distribution, and reproduction in any medium, provided the original work, first published in this journal, is properly cited

Development and validation of a sensitive, specific and reproducible uplc-ms/ms method for the quantification of ojt007, a novel anti-leishmanial agent: Application to a pharmacokinetic study

OJT007 is a methionine aminopeptidase 1 (MetAP1) inhibitor with potent anti-proliferative effects against Leishmania Major. In order to study its pharmacokinetics as a part of the drug development process, a sensitive, specific, and reproducible ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was developed and validated. Voriconazole was used as the internal standard to generate standard curves ranging from 5 to 1000 ng/mL. The separation was achieved using a UPLC system equipped with an Acquity UPLC BEH C18 column (2.1 × 50 mm, 1.7 µm) with 0.1% formic acid in acetonitrile and 0.1% formic acid in water as the mobile phase under gradient elution at a flow rate of 0.4 mL/min. The mass analysis was performed with a 4000 QTRAP® mass spectrometer using multiple-ion reaction monitoring (MRM) in the positive mode, with the transition of m/z 325 → m/z 205 for OJT007 and m/z 350 → m/z 101 for voriconazole. The intra-and inter-day precision and accuracy were within ±15%. The mean extrac-tion recovery and the matrix effect were 95.1% and 7.96%, respectively, suggesting no significant matrix interfering with the quantification of the drug in rat plasma. This study was successfully used for the pharmacokinetic evaluation of OJT007 using the rat as an animal model

Pre-clinical pharmacokinetics, tissue distribution and physicochemical studies of CLBQ14, a novel methionine aminopeptidase inhibitor for the treatment of infectious diseases

Introduction: CLBQ14, a derivative of 8-hydroxyquinoline, exerts its chemotherapeutic effect by inhibiting methionine aminopeptidase (MetAP), the enzyme responsible for the post-translational modification of several proteins and polypeptides. MetAP is a novel target for infectious diseases. CLBQ14 is selective and highly potent against replicating and latent Mycobacterium tuberculosis making it an appealing lead for further development. Methods: The physicochemical properties (solubility, pH stability and lipophilicity), in vitro plasma stability and metabolism, pre-clinical pharmacokinetics, plasma protein binding and tissue distribution of CLBQ14 in adult male Sprague-Dawley rats were characterized. Results: At room temperature, CLBQ14 is practically insoluble in water (80 mg/mL); it has a log P value of 3.03 ± 0.04. CLBQ14 exhibits an inverse Z-shaped pH decomposition profile; it is stable at acidic pH but is degraded at a faster rate at basic pH. It is highly bound to plasma proteins (\u3e91%), does not partition to red blood cells (B/P ratio: 0.83 ± 0.03), and is stable in mouse, rat, monkey and human plasma. CLBQ14 exhibited a bi-exponential pharmacokinetics after intravenous administration in rats, bioavailability of 39.4 and 90.0%, respectively from oral and subcutaneous route. We observed a good correlation between predicted and observed rat clearance, 1.90 ± 0.17 L/kg/h and 1.67 ± 0.08 L/kg/h, respectively. Human hepatic clearance predicted from microsomal stability data and from the single species scaling were 0.80 L/hr/kg and 0.69 L/h/kg, respectively. CLBQ14 is extensively distributed in rats; following a 5 mg/kg intravenous administration, lowest and highest concentrations of 15.6 ± 4.20 ng/g of heart and 405.9 ± 77.11 ng/g of kidneys, respectively, were observed. In vitro CYP reaction phenotyping demonstrates that CLBQ14 is metabolized primarily by CYP 1A2. Conclusion: CLBQ14 possess appealing qualities of a drug candidate. The studies reported herein are imperative to the development of CLBQ14 as a new chemical entity for infectious diseases

Temporal trends and black-white disparity in mortality among hospitalized persons living with HIV in the United States

We sought to determine whether black-white gap in mortality exists among hospitalized HIV-positive patients in the United States (US). We hypothesized that in-hospital mortality (IHM) would be similar between black and white HIV-positive patients due to the nationwide availability of HIV services.Our analysis was restricted to hospitalized HIV-positive patients (15-49 years). We used the National Inpatient Sample (NIS) that covered the period from January 1, 2002 to December 31, 2014. We employed joinpoint regression to construct temporal trends in IHM overall and within subgroups over the study period. We applied multivariable survey logistic regression to generate adjusted odds ratios (OR) and 95% confidence intervals (CI).The total number of HIV-related hospitalizations and IHM decreased over time, with 6914 (3.9%) HIV-related in-hospital deaths in 2002 versus 2070 HIV-related in-hospital deaths (1.9%) in 2014, (relative reduction: 51.2%). HIV-related IHM among blacks declined at a slightly faster rate than in the general population (by 56.8%, from 4.4% to 1.9%). Among whites, the drop was similar to that of the general population (51.2%, from 3.9% to 1.9%). Although IHM rates did not differ between blacks and whites, being black with HIV was independently associated with a 17% elevated odds for IHM (OR = 1.17; 95% CI = 1.11-1.25).In-hospital HIV-related deaths continue to decline among both blacks and whites in the US. Among hospitalized HIV-positive patients black-white disparity still persists, but to a lesser extent than in the general HIV population. Improved access to HIV care is a key to eliminating black-white disparity in HIV-related mortality